British Pharmacopoeia 2022 BP 2022 - VOL 1

British Pharmacopoeia 2022
Volume I
The British Pharmacopoeia Commission has caused this British
Pharmacopoeia 2022 to be prepared under regulation 317 (I) of the Human
Medicines Regulations 2012 and, in accordance with regulation 317(4), the
Ministers have arranged for it to be published.
The monographs of the Tenth Edition of the European Pharmacopoeia
(2019), as amended by Supplements 10.1 to 10.5, published by the Council
of Europe are reproduced either in this edition of the British
Pharmacopoeia or in the associated edition of the British Pharmacopoeia
(Veterinary).
See General Notices
Effective date: I January 2022
see Notices
London: The Stationery Office
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In respect of Great Britain:
THE DEPARTMENT OF HEALTH AND SOCIAL CARE

In respect of Northern Ireland:
THE DEPARTMENT OF HEALTH (NI)
© Crown Copyright 2021
Published by The Stationery Office on behalf of the Medicines and
Healthcare products Regulatory Agency (MHRA) except that:
European Pharmacopoeia monographs are reproduced with the permission
of the Council of Europe and are not Crown Copyright. These are
identified in the publication by a chaplet of stars.
This publication is a 'value added' product, If you wish to re-use the
Crown Copyright material from this publication, applications must be made
in writing, clearly stating the material requested for re-use, and the purpose
for which it is required. Applications should be sent to: Me J Pound,
MHRA, lOth Floor, 10 South Colonnade, Canary Wharf, London
EI44PU.
First Published 2021
ISBN 978 011 3230 877
British Pharmacopoeia Commission Office:
Medicines and Healthcare products Regulatory Agency
10 South Colonnade,
Canary Wharf,
London EI4 4PU
Telephone: +44 (0)203080 6561
E-mail: bpcom@mhra.gov.uk
Web site: http://www.pharmacopoeia.com
Laboratory:
British Pharmacopoeia Commission Laboratory
Queen's Road
Teddington
Middlesex TWll OLY
Telephone: +44 (0)20 8943 8960
E-mail: bpcrs@mhra.gov.uk
Web site: hnp:llwww.pharmacopoeia.com
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Foreword
The global COVID-19 pandemic has been ongoing for over a year,
drastically affecting our lives whilst unparalleled effort, innovation and
collaboration has been undertaken to respond to the crisis. This work has
included the incredibly rapid development and deployment of vaccines and
treatments by the life sciences industry and health services globally.
Whilst our focus must, and will, remain on working to support the
immediate needs of patients and the healthcare system, we must also
continue to think about a post pandemic world. It is a world where patients
will expect a progressive, responsive and pragmatic regulatory system that
facilitates scientific innovation, enables accelerated access to new medicines
and strengthens already high standards of safety, efficacy, and quality.
Standards have a vital role in this system as they are both an integral
component of ensuring medicine quality and a powerful enabler of
innovation.
I am proud to report that, throughout the pandemic, the British
Pharmacopoeia has continued to progress work to explore and reimagine
standards for the future. This includes the publication of standards on the
application of Analytical Quality by Design (AQbD), flow cytometry and
vector copy number quantification for the cell and gene therapy community.
These standards are at the forefront of a new model for how
pharmacopoeial standards can support enhanced understanding of
medicines quality and act as enablers of new technologies throughout the
product lifecycle.
The development and publication of these standards is only possible
through deep collaborations with national and international partners,
together with innovative ways of working. One such example is the
development of a staff exchange programme between the British
Pharmacopoeia and the Cell & Gene Therapy Catapult, which not only
built capability across the system but also drove forward the development of
enabling standards.
I would therefore like to thank all our staff, advisers and partners for their
expertise, dedication and flexibility throughout this unprecedented year.
Stephen Lightfoot
Chairman
Medicines and Healthcare products Regulatory Agency
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Contents
Contents of Volume I
FOREWORD
NOTICES
PREFACE
BRITISH PHARJVIACOPOEIA COMMISSION
EXPERT ADVISORY GROUPS, PANELS OF EXPERTS AND
WORKING PARTIES
CODE OF PRACTICE
MEMBERSHIP
BP Commission, Expert Advisory Groups, Panels of Experts, Working
Parties, Ad-hoc Group
STAFF
British Pharmacopoeia, BP Laboratory, Publisher
INTRODUCTION
Additions, Omissions, Technical Changes, Changes in Title
GENERAL NOTICES
MONOGRAPHS
Medicinal and Pharmaceutical Substances (A - I)
Contents of Volume II
NOTICES
GENERAL NOTICES
MONOGRAPHS
Medicinal and Pharmaceutical Substances a- Z)
Contents of Volume III
NOTICES
GENERAL NOTICES
MONOGRAPHS
Formulated Preparations: General Monographs
Formulated Preparations: Specific Monographs
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Contents of Volume IV
NOTICES
GENERAL NOTICES
MONOGRAPHS
Herbal Drugs, Herbal Drug Preparations and Herbal Medicinal Products
Materials for use in the Manufacture of Homoeopathic Preparations
Blood-related Products
Immunological Products
Radiopharmaceutical Preparations
Surgical Materials
Contents of Volume V
NOTICES
GENERAL NOTICES
INFRARED REFERENCE SPECTRA
APPENDICES
SUPPLEMENTARY CHAPTERS
INDEX
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Notices
Monographs of the European Pharmacopoeia are distinguished by a chaplet

of stars against the title. The term European Pharmacopoeia, used without
qualification, means the Tenth Edition of the European Pharmacopoeia
comprising, unless otherwise stated, the main volume, published in 2019, as
amended by any subsequent supplements and revisions.
Patents In this Pharmacopoeia certain drugs and preparations have been included
notwithstanding the existence of actual or potential patent rights. In so far
as such substances are protected by Letters Patent their inclusion in this
Pharmacopoeia neither conveys, nor implies, licence to manufacture.
Effective dates New and revised monographs of national origin enter into force on
1 January 2022. The monographs are brought into effect under regulation
320(2) of the Human Medicines Regulations 2012.
Monographs of the European Pharmacopoeia have previously been
published by the European Directorate for the Quality of Medicines &
HealthCare, in accordance with the Convention on the Elaboration of a
European Pharmacopoeia, and have been brought into effect under the
Human Medicines Regulations 2012, as amended, and the Veterinary
Medicines Regulations 2013, as amended.
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Preface
The British Pharmacopoeia Commission has caused this British

Pharmacopoeia 2022 to be prepared under regulation 317(1) of the Human
Medicines Regulations 2012 and, in accordance with regulation 317(4), the
Ministers have arranged for it to be published.
The British Pharmacopoeia 2022 contributes significantly to the quality
control of medicinal products for human use. It contains publicly available,
legally enforceable standards that provide an authoritative statement of the
quality that a product, material or article is expected to meet at any time
during its period of use. The pharmacopoeial standards are designed to
complement and assist the licensing and inspection processes and are part
of the overall system for safeguarding the health of purchasers and users of
medicinal products in the UK.
The British Pharmacopoeia also has an important role to play
internationally, being used across the globe and referenced in the national
legislation of several countries.
The British Pharmacopoeia Commission wishes to record its appreciation of
the services of all those who have contributed to this important work.
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British Pharmacopoeia
Commission
The British Pharmacopoeia Commission is appointed, on behalf of the

Secretary of State for Health and Social Care, by the Department of Health
and Social Care's Public Appointments team who are responsible for
appointments to all of the Advisory Bodies appointed under the Human
Medicines Regulations 2012.
Under the terms of the Human Medicines Regulations 2012, the duties of
the British Pharmacopoeia Commission are as follows:
(a) the preparation and publication of any new edition of the British
Pharmacopoeia [regulations 317(1) and 317(4));
(b) the preparation and publication of any compendium containing
information relating to substances and articles which are or may be
used in the practice of veterinary medicine or veterinary surgery
[regulations 317(3)(b) and 317(4));
(c) the preparation and publication of a list of names to be used as the
headings to monographs in the British Pharmacopoeia [regulations 318
(1) and 318(2));
(d) the preparation of any amendments to the above publications
[regulation 317(5)(a)).
Members of the British Pharmacopoeia Commission are appointed for a

renewable term of 4 years and, under the requirements laid down by the
Office of the Commissioner for Public Appointments, can serve for a
maximum of 10 years.
In order to ensure that the British Pharmacopoeia Commission fulfils its

duties under the Human Medicines Regulations 20 I 2, the members also
have the following duties:
(I) to frame clear and unequivocal technical advice in order to discharge
the Commission's responsibilities both for the British Pharmacopoeia,
the British Pharmacopoeia (Veterinary) and British Approved Names
and as the national pharmacopoeial authority with respect to the
European Pharmacopoeia;
(2) to develop clear policies for the preparation and publication of the
British Pharmacopoeia and its related publications;
(3) to serve on one or more Expert Advisory Groups or Panels of Experts
of the BP Commission, usually in the position of Chair or Vice-Chair;
(4) to approve new and revised text for inclusion in new editions of the
British Pharmacopoeia and British Pharmacopoeia (Veterinary);
(5) to approve new and revised names for inclusion in new editions of
British Approved Names and its annual supplements.
In addition to the duties listed above, the Chair of the British

Pharmacopoeia Commission has the following additional duties:
(1) To chair all scheduled and unscheduled meetings;
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(2) To carry out members appraisals in accordance with policies and
timelines laid down by the Department of Health and Social Care;
(3) To participate in the process to appoint/re-appoint members of the
British Pharmacopoeia Commission.
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Expert Advisory Groups, Panels
of Experts and Working Parties
Members of Expert Advisory Groups, Panels of Experts and Working

Parties are appointed by tbe British Pharmacopoeia Commission.
The duties of tbe members are as follows:

(a) to collaborate in tbe preparation and revision of Monographs,
Appendices and Supplementary Chapters for inclusion in tbe British
Pharmacopoeia and British Pharmacopoeia (Veterinary);
(b) to collaborate in tbe preparation and revision of Monographs, Metbods
and General Chapters of tbe European Pharmacopoeia;
(c) to review reports from tbe British Pharmacopoeia Laboratory in terms
of technical content and, where possible, provide independent
experimental data to assist in decision making;
(d) to collaborate in tbe preparation and revision of tbe list of names to be
used as titles for monographs of tbe British Pharmacopoeia and British
Pharmacopoeia (Veterinary).
Members of Expert Advisory Groups, Panels of Experts and Working
Parties are usually appointed for a renewable term of 4 years.
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Code of Practice
Members of the British Pharmacopoeia Commission and its supporting

Expert Advisory Groups, Panels of Experts and Working Parties are
required to comply with a Code of Practice on Declaration of Interests in
the Pharmaceutical Industry.
British Phannacopoeia Commission
The Chair and members of the British Pharmacopoeia Commission are
required to make a full declaration of interests on appointment and
annually thereafter. They must also inform the BP Secretariat promptly of
any changes to these interests during the year. These interests are published
in the Medicines Advisory Bodies Annual Reports.
Relevant interests must be declared at meetings and are recorded in the
Minutes.
Expert Advisory Groups, Panels of Experts and Working Parties
Chairs and members are required to make a full declaration of interests on
appointment and to update the Secretariat if these interests change during
their term of office. A record is kept of those experts who have declared
specific interests, but these are not published.
Relevant interests must be declared at meetings and are recorded in the
Minutes.
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Membership of the British
Pharmacopoeia Commission
The list below includes those members who served during the period 2020
to 2021.
Chair Professor Kevin M G Taylor BPharm PhD FRPharmS

Professor of Clinical Pharmaceutics, UCL School of Pharmacy
Vice-Chair Professor Alastair G Davidson BSc PhD FRPharmS

Visiting Professor of Pharmaceutical Sciences, University of Strathclyde
Dr Emre Amirak BSc MBBS MRCS
Country Medical Director UK, Ireland & Nordics, Akcea Therapeutics
Dr Andrew Barnes BSc PhD FRSC
QualityAssurance Pharmacist, Pharmacy Manufacturing Unit, East Suffolk and
North Essex NHS Trust
Dr Jon Beaman BSc PhD MBA CChem MRSC
Head of Development Analytical Group, Pfizer UK
Dr Anna-Maria Brady BSc PhD
Former Head of Biologicals and Administration, Veterinary Medicines Directorate
Dr Graham D Cook BPharm PhD MRPharmS
Senior Director, Process Knowledge/Quality by Design, Pfizer
Dr Alison Gleadle BSc PhD (Lay member)
Former Group Product Risk Director, Tesco Stores Ltd.
Dr Vikas jaitely BPharm MPharm PhD MRPharmS GPhC MTOPRA
Director (EU Digital Healthcare & Devices), Global Regulatory Affairs, Merck
Mr Robert Lowe BPharm FRPharmS
Director of Pharmacy Quality Assurance Specialist Services, NHS East of
England & Northampumshire
Dr Paul Marshall BPharm PhD MRPharmS MAPS FTOPRA
Director, Global Regulatory Affairs, Jazz Pharmaceuticals
Professor John Miller MSc PhD MRSC CChem
Visiting Professor, Strathclyde Institute of Pharmacy and Biomedical Sciences;
former Head of the EDQM Laboratory
Ms Sharon Palser MSc (Lay member)
Former Director of Development, NHS Plymouth
Professor Monique Simmonds OBE JP BSc PhD FLS FBS FRES FWIF
Deputy Director of Science, Royal Botanic Gardens, Kew
Dr Ronald Torano BSc PhD MRSC CChem
Pharmacopoeial Intelligence and Advisory Specialist; GlaxoSmithKline
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Dr Paul Varley BSc PhD
Vice President of Biopharmaceutical Development, Kymab Limited
Secretary and Me James Pound BSc

Scientific Director Group Manager, British Pharmacopoeia and Laboratory Services, MHRA
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Membership of Expert Advisory
Groups, Panels of Experts and
Working Parties
The Commission appointed the following Expert Advisory Groups, Panels

of Experts and Working Parties to advise it in carrying out its duties.
Membership has changed from time to time; the lists below include all who
have served during the period 2020 to 2021.
EXPERT ADVISORY GROUPS

ABS: Antibiotics R L Horder (Chair), G Cook (Vice-Chair), G Blake, G Clarke, E Flahive,
V [aitely, W Mann, J Miller, M Pires, J Sumal, I R Williams
BIO: Biological and P Varley (Chair), A-M Brady (Vice-Chair), E Amirak, L Bisset', C
Biotechnological Braxton" C Bums, K Chidwick', A Cook I, J Cook', B Cowper, S Gill,
Products C Jones" A Kippen, V Loh, K Nordgren', B Patel, A M Pickett',
T Pronce', L Randon, I Rces', S Schepelmanrr', P Sheppard, P Stickings",
R Thorpe, L Tsang, M Wadhwa', W Zunic
HCM: Herbal and M Simmonds (Chair), R Middleton (Vice-Chair), L A Anderson,

Complementary P Anderson, A Booker, C Etheridge, C Leon, B Moore, M Pires, E Reich,
Medicines M Rowan, A Slater, K Strohfeldt-Venables, J Sumal', C Welham,
E Williamson, K Zhao
(Corresponding members SS Handa, A Krauss, Z-T Wang)
MCI: Medicinal A G Davidson (Chair), D Cairns (Vice-Chair), S Bale, H Batchelor,

Chemicals J C Berridge, E Bush, A J Caws, D Deutsch, P Fleming, E Gray,
W J Lough, D Malpas, P Marshall, S Nolan
MC2: Medicinal G Cook (Chair), C T Goddard (Vice-Chair), J Birchall, K Boon, J Cowie, K

Chemicals Foster, E Hook, J Lim, J Miller, A Ruggiero, N Wynne
(Corresponding members M Brits, W Sherwin)
MC3: Medicinal M Almond (Chair), J Beach (Vice-Chair), J Beaman, K Foster,

Chemicals C T Goddard, P Hampshire, W K L Pugh, B Rackstraw, R Torano,
I R Williams
NOM: J K Aronson (Chair), A McFarlane, D Mehta, G P Moss, R Thorpe

Nomenclature
(Corresponding member R G Balocco Mattavelli)
PCY: Pharmacy R L Horder (Chair), R Lowe (Vice-Chair), M Ahmed', E Baker, J Beach,

D Elder, J Lim', J MacDonald, A McFarlane, J F McGuire, T Purewal,
K M G Taylor, S Wicks
(Corresponding member J Churchill)
I Specialist member.
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ULM: Unlicensed M G Lee (Chair), V Fenton-May (Vice-Chair), A Barnes, A Bosley, M
Medicines Godber, W Goddard, S Hartley, D Kirby, J Ramada-Magalhaes,
M Santillo, J Smith, A Sully, P Weir, M Westwood
PANELS OF EXPERTS
BLP: Blood K Chidwick, A R Hubbard, J More, P Varley
Products
CX: Excipients C Mroz (Vice-Chair), H Batchelor, R Cawthorne, D Deutsch
IGC: Inorganic and C T Goddard (Chair), M Almond, S Boland, P Henrys, G Lay

General Chemicals
MIC: Microbiology V Fenton-May (Chair), B Alexander, C Iverson, V Iaitely, J Silva
RAD: Radioactive I Boros, J Brain, D Graham, G Inwards, R D Pickett

Materials
VET: Veterinary E Williamson (Chair), A Cairns, S Cockbill, D Evans, E Flahive, B Ward

Medicines
VIP: Veterinary A M Brady (Chair), R Banks, R Cooney, M Ilort, M Johnson, K Redhead,

Immunological J Salt, C Stirling, R Woodland
Products
WORKING PARTIES
AQbD: Analytical G Cook (Chair), P Borman, S Brown, M Chatfield, S Ellison, C Gray,
Quality by Design M Hanna-Brown, S Jones, P Nethercote, E Razzano
(Corresponding members K Barnett, B Harrington, W Sherwin)
ATMP: Advanced J Barry (Chair), E Abranches, C Blue, J Campbell, D Caulfield, R Cowell',

Therapy Medicinal K Gilmour, J Glassford, A Lovatt, A Niewiarowska, J Norton, A Nowocin,
Products L Pattenden, J Rattu, I Rees, R Rego, V Robertson, I Santeramo, F
Schnetzinger, I Searing, B Surmacz-Cordle, J Towler, C Trento, S Vinter,
Y Zhao
NOTE: The membership incorporales that of the sub-groups on Flow Cytomeuy
and Vector Copy Number.
BIO-DPS: P Varley (Chair), A-M Brady (Vice-Chair), C Burns, B Cowper, L Duhau,

Documentary and V Ganeva, C E Giartosio, A Ramzan, B Rellahan, M Wild
Physical Standards*
* BIO-DPS: Alternative Approaches for Documentary and Physical Standards
for Biotechnological Products
AD-HOC GROUP
New Analytical J Beaman, G Cook, J Miller, M Simmonds, R Torano
Technologies
1 Deceased.
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Current British Pharmacopoeia
Staff
Secretariat J Pound (Secretary and Scientific Director)
A Gibb (Editor-in-Chief)
S Young (Head of Analytical Science)
H Ashraf, H Bowden, H Corns, P Crowley, L Elanganathan, A Evans,
G Kemp, G Li-Ship, S Maddocks, R Smith, F J Swanson, A Thomson,
M Whaley
Administrative N Begum, F Chughtai, B F Delahunty, J Paine, U Rothna
Secondees from the M Francois, R McCoy

Cell and Gene
Therapy Catapult
ISO 9001
F527268
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Current British Pharmacopoeia
Laboratory Staff
I Reydellet (Operations Manager)

D Ballottin, 0 Bennett, 0 Bernabe, C Bernardi, A Biesenbruch, M
Boardman, K Busuttil, S Choudhury, A Ciesluk, E Couzins, C Cropley, Y
EI Dabh, B Federer, S Ganguli, M Goode, S Greatorex, R Griffiths, B
Heerschop Kenalernang, D Holcombe, A Iyawe, L Magee, K Meyer de
Figueiredo, W Mohammed, G Naar, M Nanasi, A Paul, M Petrova, L
Piare, R Ravishankar, D Rutty, M Sciberras, G Searle, C Smart, C
Thompson, V Vekereya
150 9001
F527613
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Current Staff of the Publisher of
the British Pharmacopoeia
A Prince (Business Director)

P Allard (Service Delivery Manager)
N Billington, C Cole, A Dampier, C Gaines, N Griffiths, A Hughes,
N joisa, J Khurana, N Pope, M Rainbird, T Wheeler
ISO 9001
F522428
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2022 Introduction I-xxiii
Introduction
British Pharmacopoeia 2022

The British Pharmacopoeia 2022 supersedes the British Pharmacopoeia
2021. It has been prepared by the British Pharmacopoeia Commission, with
the collaboration and support of its Expert Advisory Groups, Panels of
Experts and Working Parties and contains approximately 4000 monographs
for substances, preparations and articles used in the practice of medicine.
Some of these monographs are of national origin and have been elaborated
or revised under the auspices of the British Pharmacopoeia Commission
whilst others (indicated to users by a chaplet of stars) have been elaborated,
or revised, under the auspices of the European Pharmacopoeia
Commission, supported by its Groups of Experts and Working Parties, and
are reproduced from the European Pharmacopoeia. This edition, together
with its companion volume, the British Pharmacopoeia (Veterinary) 2022,
incorporates all the monographs of the lOth Edition of the European
Pharmacopoeia, as amended by Supplements 10.1 and 10.5. Users of the
British Pharmacopoeia thereby benefit by finding within this
comprehensively indexed compendium all current United Kingdom
pharmacopoeial standards for medicines for human use.
The BP 2022 comprises six volumes as follows.
Volumes I and II Medicinal Substances

Volume III Formulated Preparations: General Monographs
Volume IV Herbal Drugs, Herbal Drug Preparations and
Herbal Medicinal Products
Materials for use in the Manufacture of
Homoeopathic Preparations
Blood-related Products
Immunological Products
Surgical Materials
Volume V Infrared Reference Spectra
Appendices
Supplementary Chapters
Index
Volume VI British Pharmacopoeia (Veterinary) 2022
Effective Date The effective date for British Pharmacopoeia monographs in this edition is
1 January 2022.
National monographs omitted from this or earlier editions of the British
Pharmacopoeia remain effective in accordance with Regulation 252(2)(c) of
the Human Medicines Regulations 2012, as amended.
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I-xxiv Introduction 2022
Implementation dates regarding European Pharmacopoeia publications are

provided in Supplementary Chapter IV B: Dates of Implementation.
European Pharmacopoeia monographs are identified by a chaplet of stars
alongside the title.
Additions A list of monographs included for the first time in the British
Pharmacopoeia 2022 is given at the end of this introduction. It includes 20
new monographs of national origin and 38 new monographs reproduced
from the 10th Edition of the European Pharmacopoeia, as amended by
Supplements 10.1 to 10.5.
The British During the coronavirus (COVID-19) outbreak the British Pharmacopoeia
Pharmacopoeia and has committed to keeping its users updated and to supporting the wider
coronavirus healthcare response. As part of this the British Pharmacopoeia has
prioritised the continued availability of written and physical standards, while
also contributing staff and expertise to the Medicines and Healthcare
products Regulatory Agency (MHRA) and participated in international
pharmacopoeial initiatives. Availability of British Pharmacopoeia standards
has been extended and expanded, including the free access publication of
relevant supportive pharmacopoeial texts in cooperation with the European
Pharmacopoeia. This has ensured that those developing, manufacturing and
testing medicines in response to the COVID-19 pandemic have had ready
access to the standards required.
Information about the British Pharmacopoeia's response to COVID-19 is
available on a dedicated webpage: https://www.pharmacopoeia.comlcovidI9.
British The BP continues to be part of the Medicines and Healthcare products

Pharmacopoeia Regulatory Agency's public health role.
Operations from 1st
January 2021 The UK was a founding member of the Convention on the Elaboration of a
European Pharmacopoeia and continues to be a member of the European
Pharmacopoeia (Ph. Eur.). The UK will continue to be a member of the
Council of Europe (CoE) in its own right. The British Pharmacopoeia (BP)
continues to reproduce Ph. Eur. text for the convenience of our customers.
In the absence of European Pharmacopoeia standards, Directive
2001l83IEC allows the continued applicability of BP standards (as a third
party pharmacopoeia) for medicines and their components. Where the BP is
appropriately referenced in the regulations of an EU member state, it may
be considered a national pharmacopoeia of that EU member state.
Previous references to EU legislation have been revised to rellect the
appropriate UK legislation. The Preliminary texts and General Notices Part
II have been revised. Several Appendices and Supplementary Chapters have
also been revised; these are listed later in this Introduction.
Pharmacopoeial The MHRA has continued to implement its strategy for pharmacopoeial
Public Quality public quality standards for biological medicines as published in 2017 and
Standards for updated in 2019 1. The strategy acknowledges the importance of biological
Biological medicines, the value of pharmacopoeial public quality standards and the
Medicines unique position of the MHRA to lead in this field through its alignment of
I The straugy and work programme can befound on thefollowing webpage: hnps:l!www.gov.uk/
gwernmentlconsuJtalionslstmtegy-f(N'-phannawpoeial-public-qualiry-standards-fo,...tn"oIogiaJ/-medicines.
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2022 Introduction I-xxv
regulatory, documentary (BP) and physical (NIESC) standard setting

functions.
Part of the published strategy was to investigate alternative approaches to
standards for biological medicines. This led to the establishment of the
Alternative Approaches for Documentary and Physical Standards for
Biotechnological Products Working Party (WP BIO-DPS) in 2018. WP
BlO-DPS has continued to develop a deeper understanding of performance
and class-based standards and consequently how documentary and physical
standards may need to evolve. These concepts are currently being evaluated
through real-world case studies coupled with supporting laboratory
assessment.
The strategy also highlighted the need to investigate and take forward
documentary and physical standard-setting opportunities for Advanced
Therapy Medicinal Products (ATMPs). The MHRA has engaged with
groups across the ATMP community to improve its understanding of the
challenges faced by those developing, manufacturing, testing and
administering these medicines and the role standards can have in supporting
innovation and assuring product quality.
The ATMP working party has developed two sets of non mandatory, best
practice guidance for the cell and gene therapy community and these have
been made available through our website. The ATMP guidelines ensure
patient safety by providing an outline of best practices to ensure product
quality is upheld throughout the product's lifecycle. The texts have been
written by experts in the flow cytometry and gene therapy community, and
are intended to be helpful to a range of stakeholders including those
operating in GMP regulated environments, research and development,
academia and clinical trials. Following the success of this work the ATMP
Working Party will identify and develop guidance on further topics that will
support the development of these innovative medicines.
Analytical QualIty The British Pharmacopoeia, working with the MHRA and stakeholders,
by Design (AQbD) continues to investigate the application of Quality by Design principles to
analytical methods and the pharmacopoeia. Several AQbD concepts have
been assessed practically in conjunction with the British Pharmacopoeia
Commission Laboratory, and the Australian Therapeutic Goods
Administration. The MHRA published the outcomes of these studies with
an accompanying public consultation I.
Consultation responses underscored the importance of AQbD concepts as
potentially transformative catalysts for enabling innovation for analytical
methods and ultimately further supporting the assurance of medicines
quality. The result of the consultation has been the adoption of a strategy
and accompanying work programme that will continue to drive forward this
important area of regulatory science. The first outcome of this work
programme, Supplementary Chapter on the use of Analytical Quality by
Design concepts for analytical procedures, is included within this
publication. The selective guidance this Supplementary Chapter provides
will support users in the application of Analytical Quality by Design
1 The consultation can befound on lkfollowing webpage: https:llwww.gov.uklgovemmemlamsultauonsl

consull<J~Wn-on-the-applialrion-of-anaJytiaJl-qua/;t;Y-by-design-tUlbd-pn·nciples-tQ-phannacqpoe; al-sta ndards
for-medicines.
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I-xxvi Introduction 2022
principles to pharmacopoeial procedures and across the entire Analytical

Method Lifecycle.
In recognition of the importance of alignment between regulators,
pharmacopoeias and stakeholders, a joint online workshop between the
MHRA and United States Pharmacopeia on pharmacopoeial application of
AQbD and Analytical Method Lifecyle concepts was held in February
2021. The workshop included speakers from industry, ICH Expert Working
Groups, the MHRA and United States Pharmacopeia.
Traditional Herbal One new British Pharmacopoeia monograph for herbal medicine is included
Medicines; in this edition (Tinospora Stem). This reflects a continued commitment to
Homoeopathic providing quality standards for herbal drugs commonly used in the UK and
Preparations for those known to be used for the preparation of traditional medicines.
The Herbal and Complimentary Medicines Expert Advisory Group has
reviewed the work programme and will continue to develop useful standards
that add value to users.
Unlicensed With this new edition, a further four monographs for unlicensed
Medicines formulations have been added. All monographs for such formulations are
characterised by a statement that the monograph has been prepared to
cover unlicensed formulations. The general and individual monographs are
intended to apply to all types of Unlicensed Medicines, that is, those
formulations prepared under a Manufacturer's 'Specials' Licence and those
prepared extemporaneously under the supervision of a pharmacist.
The Supplementary Chapter on the Aseptic Preparation of Unlicensed
Medicines 01 F) has been updated to include a new section on Ready-to-
Administer Injections. Such products are widely available and may be used
in the home environment. They are prepared in aseptic preparation units
and ate stored in a ready-to-administer form until administered to the
patient.
New Analytical LCIUV-DAD (Diode Array Detection), also known as a photo-diode array
Technologies (PDA) detection, has been introduced as a routine identification test option
in BP monographs in the BP 2022. This follows from a positive response to
a change proposal made available via the regular review schedule for draft
texts.
Revisions A significant number (130, comprising 120 technical revisions and 10

editorial revisions) of national monographs have been amended by means of
this edition. Of these monographs, those with major technical revisions are
listed at the end of this Introduction. For the benefit of the reader this list
indicates the section, or sections, of each monograph which haslhave been
revised.
The list of revisions appended to this Introduction is as comprehensive as
practicable. However, to ensure that the reader uses the current standard, it
is essential to refer to the full text of each individual monograph.
For those texts reproduced from the European Pharmacopoeia, the
European Directorate for the Quality of Medicines & HealthCare (EDQM)
database (see below, under Websites) provides information on revisions of
www.webofpharma.com
Introduction T-xxvii
the monographs or other texts on a historical basis, beginning from the 5 th

Edition of the European Pharmacopoeia.
British The British Pharmacopoeia continues to expand the catalogue of BPCRS

Pharmacopoeia which are essential parts of the published monographs. The catalogue
Chemical Reference currently contains over 800 items. The British Pharmacopoeia Commission
Substances Laboratory continuously strives to improve the percentage of BPCRS in
(BPCRS) stock and continues to aim, wherever possible, to make the BPCRS that
support new monographs for the BP 2022 and future editions, available for
users at the same time as the publication becomes available and ahead of
the implementation date.
Title Changes 9 monograph titles have been amended in this edition. The list of changes
is appended at the end of this Introduction.
Omissions 11 monographs have been omitted from the British Pharmacopoeia 2022.
Infrared Reference As with the previous edition, the reference spectra are placed in alphabetical
Spectra order within this edition.
Appendices Two new Appendices to harmonise with the European Pharmacopoeia were
first published in the British Pharmacopoeia 2021 in-year online updates.
These have been consolidated in the new edition as follows:
Appendix XIV C. Test for Bacterial Endotoxins (LAL Test) (Ph. Eur.
method 2.6.32);
Appendix VIII Z. Tetrabutylammonium in Radiopharmaceutical
Preparations (Ph. Eur. method 2.4.33).
The following Appendix has been revised:
Appendix XXI B. Approved Synonyms
Supplementary Four new Supplementary Chapters to harmonise with the European

Chapters Pharmacopoeia were first published in the British Pharmacopoeia 2021 in-
year online updates. These have been consolidated in the new edition as
follows:
SC IV T. Depyrogenation of Items used in the Production of Parenteral
Preparations (Ph. Eur. general text 5.1.12);
SC I N. Particulate Contamination (Ph. Eur. general text 5.17.2);
SC IV U. Multivariate Statistical Process Control (Ph. Eur. general text
5.28);
SC VII E. Methods of Pretreatment for Preparing Traditional Chinese
Drugs: General Information (Ph. Eur. general text 5.18).
A new Supplementary Chapter has been included for the BP 2022:
Supplementary Chapter on the use of Analytical Quality by Design
concepts for analytical procedures
The following Supplementary Chapters have been revised:
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I-xxviii Introduction 2022
SC II A. Changes in Monograph Titles

SC III Al. Contact Points
SC III A2. Expert Advisory Groups
SC IV A. Membership of the European Pharmacopoeia Commission
SC IV C. Certification Scheme
SC V Unlicensed Medicines
SC V F. Aseptic Preparation of Unlicensed Medicines
SC VII A. Traditional Herbal Medicines
SC VIII Materials for use in the Manufacture of Homoeopathic
Preparations
SC IX Similar Biological Medicinal Products
European Co-operation Agreement

Pharmacopoeia As a consequence of the Co-operation Agreement with the EDQM of the
Council of Europe, the British Pharmacopoeia Commission is pleased to
note the integration of European Pharmacopoeia texts for the British
Pharmacopoeia 2021 in-year online updates and for this edition of the
British Pharmacopoeia.
In accordance with previous practice, all monographs and requirements of
the European Pharmacopoeia are reproduced in this edition of the British
Pharmacopoeia or, where appropriate, within its companion edition, the
British Pharmacopoeia (Veterinary) 2022.
Where a monograph has been reproduced from the European
Pharmacopoeia, this is signified by the presence of a chaplet of stars
alongside its title. Additionally, reference to the European Pharmacopoeia
monograph number is included immediately below the title in italics in the
form 'Ph. Bur. monograph >:xxx'. Where the title in the British
Pharmacopoeia is different from that in the European Pharmacopoeia, an
approved synonym has been created (see Appendix XXI B) and the
European Pharmacopoeia title is included before the monograph number.
The entire European Pharmacopoeia text is delineated by two horizontal
lines bearing the symbol' Ph. Bur.'.
The European Pharmacopoeia texts have been reproduced in their entirety
but, where deemed appropriate, additional statements of relevance to UK
usage have been added (e.g. action and use statement, a list of British
Pharmacopoeia preparations). It should be noted, however, that in the
event of doubt of interpretation in any text of the European
Pharmacopoeia, the text published in English under the direction of the
Council of Europe should be consulted.
Correspondence between the general methods of the European
Pharmacopoeia and the appendices of the British Pharmacopoeia is
indicated in each appendix and by inclusion of a list at the beginning of the
appendices section.
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Introduction I-xxix
Pharmacopoeial It should be noted that any article intended for medicinal use which is
Requirements described by a name at the head of a monograph in the current edition of
the Pharmacopoeia must comply with that monograph 'whether or not it is
referred to as BP'.
It is also important to note that no requirement of the Pharmacopoeia can
be taken in isolation. A valid interpretation of any particular requirement
depends upon it being read in the context of (i) the monograph as a whole,
(ii) the specified method of analysis, (iii) the relevant General Notices and,
where appropriate, (iv) the relevant General Monograph(s). Familiarity with
the General Notices of the Pharmacopoeia will facilitate the correct
application of the requirements. Additional guidance and information on
the basis of pharmacopoeial requirements is provided in Supplementary
Chapter I. This non-mandatory text describes the general underlying
philosophy and current approaches to particular aspects of pharmacopoeial
control.
Code of Practice Members of the British Pharmacopoeia Commission and its supporting
Expert Advisory Groups, Panels of Experts and Working Parties are
required to comply with a Code of Practice on Declaration of Interests in
the pharmaceutical industry. Details of the Code are published on the
website (pharmacopoeia.com).
Websites British Pharmacopoeia Website

The British Pharmacopoeia website, pharmacopoeia.com, contains
information relating to the British Pharmacopoeia. It allows subscribers to
access the British Pharmacopoeia 2022 and British Pharmacopoeia
(Veterinary) 2022 online and British Approved Names publications. All
users are also able to view and purchase BPCRS products through the
website.
In-year updates to the British Pharmacopoeia 2022 will be published on the
website in order to enable users to keep up to date with texts published in
Supplements 10.6 to 10.8 of the 10th Edition of the European
Pharmacopoeia. These updates will be integrated annually with the
publication of the main edition of the British Pharmacopoeia.
Chromatograms for information to support new monographs published in
the British Pharmacopoeia 2022 have been added to the example tesr
results gallery to aid users of British Pharmacopoeia monographs. This
service will increase year-on-year to allow users to examine chromatograms
obtained during the practical evaluation of new monographs by the British
Pharmacopoeia Commission Laboratory.
A new set of pages has been added to the website which host projects which
go to public consultation for review: https://www.pharmacopoeia.com/bp-
consultations. Past and live consultations will be published here to increase
the transparency of the work of the BP and to provide an appropriate point
of contact for stakeholders to engage and respond to the work of the BP.
A regular review schedule for draft texts is included on the website, with
draft new and revised monographs being posted at the start of each quarter
and available for comment for a period of three months thereafter. This free
www.webofpharma.com
L-xxx Introduction 2022
service allows greater visibility of the BP's work programme and enables
stakeholder contributions to monograph development.
Subscribers to the BP online will find that draft texts and example test
results are also linked with relevant texts and directly accessible from the
BP online content. Additionally, BPCRS products are also linked with
relevant BP monographs and subscribers to the BP online will be able to
purchase these directly from the BP online. BPCRS customers are able to
make purchases through invoice or credit card orders.
An email subscription feature allows users to keep abreast with BP news.
Additionally, users can subscribe to receive BPCRS updates, which are now
posted monthly.
Access to previous editions of the BP is available as a BP archive product
for purchase by new and existing BP online subscribers. The content of the
archive starts from the BP 2014 onwards and grows year-on-year as
superseded editions are added to the archive.
The British Pharmacopoeia is committed to improving users' experience of
its products and services through a programme of continuous improvement
based on ongoing independent user research. This research enables
identification of user needs and the development of enhancements, several
of which have been deployed to the BP website. These enhancements
include:
• A simple guide on how to use the BP;
• Enhancement of the BPCRS catalogue enabling search by CAS number
and a record of leaflets for previous batches;
A tracked changes feature allowing users to identify what content has
been changed when texts have been revised through the addition of clear
textual mark-ups.
• A Revision History feature providing users with the justifications for
changes made to monographs between editions
European Pharmacopoeia Websites
For those texts reproduced from the European Pharmacopoeia, the EDQM
website provides access to a database (the Knowledge database: https://
www.edqm.eulenlknowledge-database) containing information of various
sorts related to monographs and intended to facilitate their proper use.
Information is provided on chromatographic columns used in monograph
development, suppliers of reagents and equipment that may be difficult to
find for some users, the status of monographs (in development, adopted,
published, under revision), revisions of the monographs on a historical
basis, beginning from the 5th Edition of the European Pharmacopoeia as
well as other useful information.
The European Pharmacopoeia Forum, Pharmeuropa, is published quarterly
as an aid for the elaboration of monographs and as a vehicle for information
on pharmacopoeial and related matters. Pharmeuropa is available as a free
online publication: https:/Ipharmeuropa.edqm.eulhome
International Therapeutic Goods Administration, Australia The British

Collaboration Pharmacopoeia Commission is pleased to continue its long-standing co-
operation with the Australian Department of Health Therapeutic Goods
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Introduction I-xxxi
Administration (TGA). The TGA continues to provide advice to British

Pharmacopoeia Commission Expert Advisory Groups, to participate in
inter-laboratory evaluation of British Pharmacopoeia monographs and to
review data jointly. This collaboration has enabled the production of robust,
high qualiry monographs for users.
Chinese Pharmacopoeia The British Pharmacopoeia Commission is
pleased to continue its collaboration with the Chinese Pharmacopoeia on
the development of monographs and staff exchanges to support mutually
agreed projects.
Croatian Agency for Medicinal Products and Medical Devices
("HALMED") The Cooperation Agreement between the Medicines and
Healthcare products Regulatory Agency and HALMED provides a licence
for the use of information in the British Pharmacopoeia on unlicensed
medicines.
Indian Pharmacopoeia A Memorandum of Understanding was signed
with the Indian Pharmacopeia in March 2021. The Memorandum of
Understanding will allow for the exchange of information on the qualiry of
medicines and technical expertise regarding the development of standards,
methods and supporting materials.
Japanese Phannacopoeia The British Pharmacopoeia has collaborated
with the Japanese Pharmacopeia for the development of informally
harmonised standards and knowledge sharing in a number of areas of
mutual interest.
State Pharmacopoeia of the Republic of Kazakhstan Following the
signing of a Collaboration Agreement in April 2016, the Medicines and
Healthcare products Regulatory Agency has granted the Committee on
Surveillance of Medical and Pharmaceutical Activities of the Ministry of
Health of the Republic of Kazakhstan a licence to continue to use relevant
contents of the British Pharmacopoeia in the State Pharmacopoeia of the
Republic of Kazakhstan.
State Pharmacopoeia of Ukraine Following the signing of a
Collaboration Agreement in 2016, the Medicines and Healthcare products
Regulatory Agency has continued to grant the Ukrainian Scientific
Pharmacopoeial Center for Qualiry of Medicines a licence to use relevant
contents of the British Pharmacopoeia in the State Pharmacopoeia of
Ukraine.
United States Pharmacopeia A Memorandum of Understanding was
signed with the United States Pharmacopeia in July 2019. The
Memorandum of Understanding builds on the success of the programme of
work to jointly develop and revise drug product monographs and will
facilitate further knowledge sharing and joint participation in conferences
and symposia in areas of mutual interest. This included the development
and hosting of a joint BP and USP webinar on Analytical Quality by Design
and Analytical Method Lifecycle concepts to the pharmacopoeia in
February 2021.
World Health Organization The collaboration agreement between the
British Pharmacopoeia and the International Pharmacopoeia continues to
support the work of the WHO, including collaboration and information
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I-xxxii Introduction 2022
exchange, contribution to the International Meeting of World

Pharmacopoeias, and the international non-proprietary names programme.
The BP has collaborated with global pharmacopoeias under the auspices of
the WHO IMWP to respond to the global COVID-19 Pandemic.
Acknowledgements The British Pharmacopoeia Commission is greatly indebted to the members

of its Expert Advisory Groups, Panels of Experts and Working Parties for
their dedicated enthusiasm and assistance in the preparation of this edition.
In particular, member's' commitment and adaptability which has enabled
continued participation in BP meetings, albeit remotely, during the
COVID-19 pandemic.
The British Pharmacopoeia Commission would also like to thank the

laboratory team who support the work of the British Pharmacopoeia, in
particular for continuing to maintain the laboratory service throughout the
pandemic.
Close co-operation has continued with many organisations in the United

Kingdom and overseas. These include the Medicines and Healthcare
products Regulatory Agency, the Veterinary Medicines Directorate, the
Royal Pharmaceutical Society, the Association of the British Pharmaceutical
Industry the British Association of Homoeopathic Manufacturers, the
United Kingdom Herbal Forum, the Cell and Gene Therapy Catapult, the
China National Medical Products Administration, the Chinese
Pharmacopoeia Commission, the European Pharmacopoeia Commission
and the European Directorate for the Quality of Medicines & HealthCare,
the Therapeutic Goods Administration (Australia), the Health Products and
Food Branch of Health Canada, the United States Pharmacopeia, the
Quality Assurance and Safety: Medicines Department of the World Health
Organization, the Health Sciences Authority of Singapore and the Royal
Botanic Gardens, Kew.
The British Pharmacopoeia Commission wishes to thank the European

Directorate for the Quality of Medicines & HealthCare for their support
and assistance in the reproduction of the European Pharmacopoeia texts
and monographs. The British Pharmacopoeia Commission acknowledges
the importance of the work of the European Pharmacopoeia Commission
and its Groups of Experts and Working Parties. The British Pharmacopoeia
Commission is also grateful for the generous contribution by the UK
experts to the work of the Groups of Experts and Working Parties of the
European Pharmacopoeia Commission.
The British Pharmacopoeia Commission also acknowledges and appreciates

the advice of the publishing-team at The Stationery Office, in particular, Me
Paul Allard, Ms Nichola Billington, Me Chris Cole, Me Ashley Dampier,
Ms Charlotte Gaines, Ms Natasha Griffiths, Me Adrian Hughes, Me
Nagaraja [oisa, Me [aspaul Khurana, Me Andrew Prince, Ms Nichol Pope,
Me Mark Rainbird, and Me Thomas Wheeler, in the production of this
edition.
The British Pharmacopoeia Commission also acknowledges the contribution

of two members of the Cell and Gene Therapy Catapult, Dr Moira
Francois and Dr Ryan McCoy who participated in a secondment to the
British Pharmacopoeia Secretariat.
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Introduction I-xxxiii
Additions The following monographs of the British Pharmacopoeia 2022 were not
included in the British Pharmacopoeia 2021.
Medicinal and Pharmaceutical Substances

Aspirin Lysine'
Deferasirox'
Ethanolamine"
Ibandronate Sodium Monohydrate!
Latanoprosr'
Pemetrexed Disodium 2.5-Hydrate l
Riociguat'
Rivaroxaban I
Sorafenib Tosilate!
Teriflunomide!
Ticagrelor'
Trifluridlne'

Intravesical Preparations I
Medicated Plasters I

Benzylpenicillin Infusion
Ciprofloxacin Ear Drops
Ciprofloxacin Hydrochloride Eye Drops
Ciprofloxacin Oral Suspension
Dronedarone Tablets l
Flucloxacillin Infusion
F1uticasone and Salmeterol Inhalation Powder
Folic Acid Oral Solution
Ibuprofen Effervescent Granules
Magnesium Sulfate, Potassium Chloride and Sodium Chloride Infusion
Mebendazole Oral Suspension
Methadone Concentrate for Oral Solution
Pregabalin Capsules
Pregabalin Oral Solution
Repaglinide Tablets
Rifaximin Tablets
Rivastigmine Capsules
Rivastigmine Oral Solution
Rivastigmine Transdermal Patches
Regorafenib Tablets!
Riociguat Tablets l
Rivaroxaban Tablets!
Sorafenib Tablets l
Ticagrelor Tablets l
Tranexamic Acid Oral Solution
Herbal Drugs, Herbal Drug Preparations and Herbal Medicinal

Products
Chaenomeles Fruit!
Cyathula Root l
Forsythia Fruit!
1 Denotes a monograph of me European Phannacopoeia.
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I-xxxiv Introduction 2022
Ganoderma'
Morinda Root'
Tinospora Stem
Betiatide for Radiopharmaceutical Preparations!
Gallium (68Ga) Chloride (Accelerator-produced) Solution for
Radiolabelling!
Gallium (68Ga) PSMA-ll Injection'
e
PSMA-1007 8F)Injection I
Omissions The following monographs of the British Pharmacopoeia 2021 are not
included in the British Pharmacopoeia 2022.
Arnobarbital''
Amobarbital Sodium"
Carisoprodol"
Colecalciferol Concentrate (Water-dispersible Form)"
Ethanclamine"
Meprobamate"
Metrifonate''
Nalidixic Acid3
Theobromine4
Trifluridine"

Biphasic Insulin Iniecrion''
Technical Changes The following monographs in the British Pharmacopoeia 2022 have been
technically amended since the publication of the British Pharmacopoeia
2021, or have had a significant editorial change. This list does not include
revised monographs of the European Pharmacopoeia. An indication of the
nature of the changers) or the section(s) of the monograph that haslhave
been changed is given in italic type in the right hand column.
Diclofenac Diethylamine Related substances

Oxytetracycline Calcium Definition; Related substances; Assay
Promethazine Teoclate Graphic formula (corrected)
Sodium Feredetate Free sodium edetate; Nitrilotriacetic acid
Sumatriptan Impurities A and H; Related substances;
Assay
Aciclovir Cream Related substances

Aciclovir Infusion Related substances
I Denotes a monograph of theEuropean Pharmacopoeia.

2 Monograph suppresud by the European Pharmacopoeia Commission on 1stApn"12021.
3 Monograph suppressed by the European Pharmacopoeia Commission on lsi January 2021.
4 lWonograph suppressed by lire European Pharmacopoeia Commi.ssion on lstJu/y 2021.
5 Replaced by Ph Bur Monograph.
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Introduction T-xxxv
Aciclovir Ointment Related substances

Aciclovir Oral Suspension Related substances
Aciclovir Tablets Related substances
Aciclovir Dispersible Tablets Related substances
A1endronic Acid and Colecalciferol Uniformity of content
Tablets
Azitbromycin Tablets Dissolution
Beclometasone Pressurised Inbalation Content of beclometasone dipropionate;
Uniformity of delivered dose; Assay;
LabeUing
Bendroflumetbiazide Tablets Content of bendroflumethiazide;
Identification; Dissolution; Related
substances; Assay; Impurities
Cefalexin Capsules Identification; Disintegration (deleted);
Dissolution; Relatedsubstances; Assay
Cefalexin Tablets Identification; Relatedsubstances
Ceftazidirne Eye Drops Definition; Acidity or alkalinity; Related
substances; Assay for sodium carbonate
(deleted); Storage
Ceftazidirne Injection Related substances
Cilastatin and Imipenem for Infusion Related substances
Cirnetidine Oral Suspension Related substances
Ciprofloxacin Infusion Related substances; Assay
Ciprofloxacin Tablets Dissolution; Related substances; Assay
Clindamycin Injection Related substances; Assay
Clobazam Tablets Dissolution
Clomipramine Capsules Related substances
Clonidine Injection Related substances
Co-codamol Capsules Dissolution; Relatedsubstances; Assay
Co-codamol Tablets Dissolution; Related substances; Assay
Co-codarnol Effervescent Tablets Identification; Relatedsubstances; Assay
Co-dydrarnol Tablets Dissolution; Related substances; Assay
Dalteparin Sodium Injection Identification testA; Assay
Diltiazem Prolonged-release Tablets Related substances
Doxorubicin Sterile Concentrate Identification
Doxorubicin For Infusion Identification
Dosulepin Tablets Related substances
Enoxaparin Sodium Injection Identification test A; Assay
Estradiol Vaginal Tablets Content of estradiol
Farnotidine Tablets Related substances
Ferrous Fumarate and Folic Acid Dissolution
Capsules
Ferrous Fumarate and Folic Acid Dissolution
Tablets
Flavoxate Tablets Related substances; 3-Methylflavone-8-
carboxylic Acid
Fluorescein Injection Related substances
Furosemide Injection Related substances
Furosemide Oral Solution Related substances
Furosemide Tablets Related substances
Fusidic Acid Cream Assay
Fusidic Acid Oral Suspension Assay
Gliclazide Tablets Dissolution; Relatedsubstances
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I-xxxvi Introduction 2022
Haloperidol Capsules Identification; Dissolution; Related

substances; Impurities
Haloperidol Injection Content of haloperidol; Identification;
Related substances; Assay; Impurities
Haloperidol Oral Solution Definition; Characteristics (deleted);
Identification; Related substances; Assay;
Impurities
Haloperidol Tablets Content of haloperidol; Dissolution;
Related substances; Impurities
Heparin Injection Identification test C
Hydroxychloroquine Tablets Definition; Content of
hydroxychloroquine sulfate;
Identification; Disintegration (deleted);
Dissolution; Related substances; Assay;
Impurities
Hyoscine Burylbromide Injection Related substances
Hyoscine Butylbromide Tablets Related substances
Ibuprofen Capsules Related substances
Ibuprofen Prolonged-release Capsules Related substances
Ibuprofen Gel Related substances
Ibuprofen Oral Suspension Related substances
Ibuprofen Tablets Related substances
Ibuprofen Orodispersible Tablets Related substances
Ibuprofen Prolonged-release Tablets Related substances
Ketoconazole Cream Related substances
Ketoconazole Shampoo Related substances
Levothyroxine Oral Solution Identification; Related substances
Lidocaine Intraocular Injection Related substances
Loperamide Capsules Related substances
Loperamide Oral Solution Related substances
Loperamide Oral Suspension Related substances
Loperamide Orodispersible Tablets Related substances
Loperamide Tablets Related substances
Melatonin Capsules Identification
Melphalan for Injection Related substances; Impurities
Methadone Injection Identification; Related substances; Assay
Methadone Oral Solution Related substances; Assay
Methadone Tablets Identification; Dissolution; Related
substances; Assay
Mexiletine Capsules Content of mexiletine hydrochloride;
Identification; Dissolution; Related
substances; Impurities
Mirtazapine Oral Solution Related substances
Mirtazapine Tablets Related substances
Mirtazapine Orodispersible Tablets Related substances
Moxonidine Tablets Related substances
Mycophenolate Mofetil Capsules Related substances
Mycophenolate Mofetil for Infusion Related substances
Mycophenolate Mofetil Oral Assay
Suspension
Mycophenolate Mofetil Tablets Related substances
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Introduction I-xxxvii
Naproxen Oral Suspension Content of naproxen; Identification;

Acidity; Dissolution; Relatedsubstances;
Assay
Naproxen Tablets Identification; Dissolution; Related
Naproxen Gastro-resistant Tablets Definition; Dissolution; Related
substances; Assay
Niclosamide Chewable Tablets Title change; Identification; 5-
Chlorosalicylic acid
Nicotine Inhalation Cartridges Related substances
Nicotine Nasal Spray Relatedsubstances; Assay
Nicotine Sublingual Tablets Related substances
Nicotine Transdennal Patches Related substances; Assay
Nicotine Resinate Medicated Related substances
Chewing Gum
Norfloxacin Tablets Identification test A; Related substances
Ondansetron Injection Relatedsubstances; Bacterial endotoxins
(deleted)
Ondansetron Tablets Definition; Dissolution; Related
substances
Oxytetracycline Capsules Dissolution; Related substances; Assay;
Impurities
Oxytetracycline Tablets Dissolution; Related substances; Assay;
Impurities
Paracetamol and Caffeine Tablets Identification test B; Dissolution for
paracetamol; Dissolution for caffeine
(deleted); Related substances
Paracetamol and Caffeine Soluble Identification; Disintegration (deleted);
Tablets Related substances
Paracetarnol, Codeine Phosphate and Identification test D; Dissolution; 4-
Caffeine Capsules Aminophenol (deleted); Related
substances; Assayfar codeine phosphate
Paracetamol, Codeine Phosphate and Identification test D; Dissolution; 4-
Caffeine Tablets Aminophenol (deleted); Related
substances; Assay far codeine phosphate
Parenteral Nutrition Solutions Preparation; Labelling
Povidone-Iodine Mouthwash Identification tests G and D
Pyridoxine Tablets Identification; Dissolution; Related
Sertraline Tablets Related substances
Sildenafil Chewable Tablets Related substances
Simvastatin Tablets Identification; Related substances
Sodium Fusidate Ointment Assay
Sodium Fusidate Tablets Assay
Sodium Valproate Prolonged-release Related substances; Assay
Capsules
Sodium Valproate Oral Solution Relatedsubstances
Sodium Valproate Tablets Dissolution; Related substances; Assay
Sodium Valproate Gastro-resistant Related substances; Assay
Tablets
Tablets
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I-xxxviii Introduction 2022
Streptomycin Injection Manograph replaced by that for

Streptomycin Sulfate for Injection;
Requirements for the ready-to-use
solution have been deleted; Identification
testA
Sumatriptan Tablets Impurities A and H; Related substances;
Assay
Tigecyeline for Infusion Related substances; Assay
Tinzaparin Sodium Injection Idemification test A; Assay
Tranexamic Acid Injection ldentification; Related substances; Assay
Trichloroacetic Acid Solution ldentification test B; Assay
Vecuronium Bromide for Injection Related substances

Products
Capsicum Tincture Assay
Changes in Title The following list gives the alterations in the titles of monographs of the
British Pharmacopoeia 2021 that have been retained in the British
Pharmacopoeia 2022.
BRITISH PHARMACOPOEIA BRITISH PHARMACOPOEIA

2021 2022
Ferrous Gluconate Ferrous Gluconate Hydrate

Piperacillin Piperacillin Monohydrate
Colloidal Silver for External Use Colloidal Silver
Sulfur for External Use Sulfur
Transderrnal Patches Patches
Nielosamide Tablets Nielosamide Chewable Tablets

Products
Quantified Hawthorn Leaf and Hawthorn Leaf and Flower Liquid

Flower Liquid Extract Extract
Passion Flower Passionflower Herb
Passion Flower Dry Extract Passionflower Herb Dry Extract
www.webofpharma.com

Acidity; Dissolution; Related substances;
Assay
substances; Assay
Chlorosalicylic acid
Nicotine Nasal Spray Related substances; Assay
Nicotine Transdermal Patches Related substances; Assay
Chewing Gum
Ondansetron Injection Related substances; Bacterial endotoxins
(deleted)
substances
Impurities
Impurities
Paracetamol and Caffeine Tablets Identification test B; Dissolution for
ParacetamoJ and Caffeine Soluble Identification; Disintegration (deleted);
substances; Assayfor codeine phosphate
Povidone-Iodine Mouthwash Identification tests C and D
Capsules
Sodium Valproate Oral Solution Related substances
Tablets
Tablets
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T-xxxviii Introduction 2022
Streptomycin Injection iV!onograph replaced by that for

Requirements for the readY-la-use
testA
Assay
Tigecycline for Infusion Related substances; Assay
Tinzaparin Sodium Injection Identification test A; Assay
Tranexamic Acid Injection Identification; Related substances; Assay
Trichloroacetic Acid Solution Identification test B; Assay

Products
Pharmacopoeia 2022.
BRITISH PHARMACOPOEIA BRITISH PHARi\1ACOPOEIA

2021 2022

Niclosamide Tablets Niclosamide Chewable Tablets

Products

Passion Flower Dry Extract PassionflowerHerb Dry Extract
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Acidity; Dissolution; Related substances;
Assay
substances; Assay
Chlarosalicylic acid
Nicotine Nasal Spray Related substances; Assay
Nicotine Transderrnal Patches Related substances; Assay
Chewing Gum
Ondansetron Injection Related substances; Bacterial endotoxins
(deleted)
substances
Impurities
Impurities
Paracetarnol and Caffeine Tablets Identification test B; Dissolution for
Paracetamol and Caffeine Soluble Identification; Disintegration (deleted);
Paracetamol, Codeine Phosphate and Identification test D; Dissolution; 4- .
Povidone-Iodine Mouthwash Identification tests C and D
Capsules
Sodium VaJproate Oral Solution Related substances
Tablets
Tablets
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I-xxxviii Introduction 2022
Streptomycin Injection Monograph replaced by that for

Requirements for the ready-to-use
test A
Assay
Tigecycline for Infusion Relatedsubstances; Assay
Tinzaparin Sodium Injection Identification test A; Ass<UI
Tranexamic Acid Injection Identification; Related substances; Assay
Trichloroacetic Acid Solution Identification test B; Assay

Products
Pharmacopoeia 2022.
BRITISH PHARMACOPOEIA BRITISH PHARMACOPOEIA· ...

2021 2022

Niclosamide Tablets Niclosamide Chewable Tablets

Products

Passion Flower Dry Extract Passionflower Herb Dry Extract
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2022 General Notices I-I
General Notices
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1-2 General Notices 2022
CONTENTS OF THEGENEIlAL NOTICES

Parll Implementation of PharmacopoeialMethods
Italic introduction Conventional Terms
EuropeanPharmacopoeia Interchangeable Methods
References to Regulatory Documents
Parln 1.2 Other Provisions Apply.ing to General Chapters
Iutlicinrroducwm . and Monographs
Official Standards Quantities
Definition of Terms Apparatus and Procedures
Expression of Standards Water_bath
Temperature Drying and Ignition to Constant Mass
Weights and Measures ·Reagents
Atomic Weights Solvents
Constant Weight Expression of Content
Expression of Concentrations Temperature
Water Bath 1.3 General Chapters
Reagents Containers
Indicators 1.4 Monographs
Caution.Statements . Tides
Tides • ..•.. . Relative Atomic and Molecular Masses
Chemical Formulae Chemical Abstracts Service (CAS) Registry
Definition . Number
Production Definition
Manufacture of Formulated Preparations Limits of Content
Freshly and Recently Prepared l-Ierbal Drugs
Methods-of Sterilisation Production
Water Choice. of Vaccine Strain, Choice of Vaccine
Excipients •..•. Composition .. . . .
Colouring Agents" Potential Adulteration.
Antimicrobial Preservatives Characters
Characteristics . .. Solubility
Solubility Identification
Identification Scope
Reference Spectra First and Second Identifications
AssaysandTests Powdered HerbalDrugs
Biological Assays and Tests .. . Tests and Assays
Reference Substances and Reference Preparations Scope
ChemicalReference. Substances Calculation
Biological Reference Preparations Limits
Storage Indication of Permitted Limit ofImpurities
Labelling Herbal Drugs
Action and Use Equivalents
Crude Drugs; Traditional Herbal and Culture Media
Complementary Medicines Storage
Monograph Tide Labelling
Definition Warnings
Characteristics Impurities
Control.Methods Functionality-related Characteristics of
HomoeopathicMedicines Excipients .
Unlicensed.Medicines Reference Standards
PartIn 1.5 Abbreviations and Symbols
Italicintroduction Abbreviations used in the Monographs on
General Notices of the European Pharmacopoeia Immunoglobulins, Irnmunosera and Vaccines
1.1 General Statements Collections. of Micro-organisms
Quality Systems 1.6 Units of the International System (SI) used in
Alternative Methods the Pharmacopoeia and Equivalence with
DcrrionstrationofCompliance with the other Units
Pharmacopoeia International System of Units(SI)
Grade of Materials Notes
General Monographs
Validation of Pharmacopoeial Methods
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2022 General Notices 1-3
Part I
. -
The British Pharmacopoeia.comprises the entire ~x(within this publication. - The

word 'official' is used in the.Pharmacopoeia to signify 'of the Pharmacopoeia'. It
appliesto any title, substance, preparation, method or statement included in the
general notices, monographs and appendices,of the Pharmacopoeia. The
'abbtwiatiotl for British Pharmacopoeia is BP.
European Monographs of the European Pharmacopoeia are reproduced in this edition

. Pharmacopoeia' of the British Pharmacopoeia by incorporation of the text published under
the direction of the Council of Europe (Partial Agreement) in accordance
with the Convention on'the Elaboration of a European Pharmacopoeia
(Treaty Series 1'10.32 (1974) CMND 5763) as amended by the Protocol to
the Convention (Treaty Series No. MISC16 (1990) CMND 1133) ..They
are included for the convenience of users of the British Pharmacopoeia. In
cases of doubt or dispute reference should be made to the Council of
Europe text." .
•'**.' Monographs of the European Pharmacopoeia are distinguished by a
.. : chaplet 'of stars against the title and by reference to the European
** * Pharmacopoeiainonograph number included immediately below the
title in italics. The beginning and end of text from the European
Pharmacopoeia ate denoted by means of hOnZontallirieswith the symbol
'Ph EUt' ranged left and right, respectively. .' .
The general provisions of the European Pharmacopoeia relating to
different .types of dosage form are included in the appropriate general
monograph in that Section of the British Pharmacopoeia entitled
Monograph~: Formulated Preparations. These general provisions apply to
all dosage forms of the type defined, whether or not. an individual -
monograph-is included in the British Pharmacopoeia. In addition, the
provisions of the European Pharmacopoeia General Monograph for
Pharmaceutical Preparations apply to all dosage forms, whether or not an
individual monograph is included in the British Pharmacopoeia.
Texts of the European Pharmacopoeia are governed by the General
. Notices of the European Pharmacopoeia. These are reproduced as Part III
of these notices.
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Part II
The following general notices applyw the statements made in the monographs of
the British Pharmacopoeia other than those reproduced. from the European
Pharmacopoeia and tothe statements made in the'Appendices of the British
-Pharmacopoeia other than iohen a method, test or other matter described in an
appendix is invoked ill a monograph reproduced from the European
Pharmacopoeia.
Official Standards The requirements stated in the monographs of the Pharmacopoeia apply to
articles that are intended for medicinal use but not necessarily to articles
that may be sold under the same name for other purposes. An article
intended for medicinal use that is described by means of an official tide
must comply with die requirements of th~ .relevantmonograph, A
formulated preparation must comply throughout its assigned shelf-life.
(period of validity). The subject of any other monograph must comply
throughout its period of use. _
A monograph is to be construed in accordance with any general
monograph or notice or any appendix, note or other explanatory material
that is contained in-this edition_ and that is applicable to that monograph.
All statements contained in the monographs, except where-a specific general
notice indicates otherwise and with the exceptions given be1ow,~onstitnte
.standards for the official articles. An article is-not of pharmacopoeial quality -
unless it complies with all of the requirements stated. This does not imply
that.a manufacturer is obliged to perform all the tests in a monograph in
order to assess compliance with the Pharmacopoeia before release of a
product. The manufacturer may assure himself that a product is of
pharmacopoeial quality by other means, for example, from d~ta derived
froni validation studiesof the manufacturing process.from in-process -
_ controls or from a combination of the two. Parametric release in
appropriate circumstances is thus not precluded by the need to comply with
the-Pharmacopoeia.- The general notice on-Assays and Tests indicates that
analytical methods,other than those described In the Pharmacopoeia may be
employed for routine purposes. _
Requirements in monographs have been framed to provide appropriate __
-limitation of potential inipurities rather than to provide against all. possible
impurities. Material found to contain an impurity not detectable by means
of the prescribed tests is not of pharmacopoeial quality if the nature or
amount of the impurity found is incompatible with good pharmaceutical
practice. , _
The status of any statement given under the headings Definition, _
Production, Characteristics, Storage, Labelling or-Action and use is defined-
within the general notice relating to the relevant heading. In addition to any
exceptions indicated by one of the general notices referred to above, the-
following parts of a monograph do not constitute standards: (a) a graphic or ,
molecular formula given at the beginning of a monograph; (b) a molecular
weight; (c) a Chemical Abstracts Service Registry Number; (d) any
information given at the end of a monograph concerning impurities known
to be limited bythat monograph; (e) information in any annex to a I
I
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I
j
2022 General Notices ~ 1-5
monograph. Any statement containing the word 'should' constitutes non-

mandatory advice or recommendation.
The expression 'unless otherwise justified and authorised' means that the
requirement jn question has to be met, unless a competent authority
authorises a modification or exemption where justified in a particular case..
The term 'competent authority' means the national, supranational or
. international body or organisation vested with the authority for making
decisions concerning the issue in question. 1£ may, for example, be a- ~
, licensing authority or an official control laboratory. For aformulated. ~
preparation that is the subject. of monograph in the British Pharmacopoeia
any ~ justified and authorised modification to, or exemption from, the'
requirements of the relevant t~neral monograph of theEuropean ~
Pharmacopoeia is stated in the individual monograph. For example, the
~ general monograph for Tablets requires that Uncoated Tablets, exceptfor
chewable tablets, disintegrate within 15 minutes; for Calcium Lactate'
Tablets a time of 30 minutes .is-permitted..
Many of the general monographs. for formulated preparations include
statementsand requirements additional to those of the European,- -
Pharmacopoeia that are applicable to theindividualmonographs of the
British Pharmacopoeia. Such statements and requirements apply to all
monographs for that dosage form included in the Pharmacopoeia unless
otherwise indicated in the individual monograph.
Where a monograph on a biological substance 'or preparation refers to a
strain, a test, a method, a substance, erc., using' the qualifications 'suitable'
or 'appropriate' without further definition in the textj-the choice of such
strain; test, method, substance, et~., is made in accordance with any
international agreements ornational regulations affecting-the subject'
concerned. ~-
Definition of Terms Where the term 'about' is includ~din. a monograph or test it should be . ~
taken to mean. approximately (fairly correct or accuratejnear to the actual
value).
Where the term 'corresponds' is included in amonograph or test it
should be taken to mean. similar or equivalent in characteror-quantity,
Where the term 'similar' is included in a monograph or test it should be
taken to mean alike though not necessarily identical.
~F!Jrtherqualifiers (such as numerical acceptance criteria) for the above
terms are not included in the BP. The acceptance criteria for any indivIdual
case is set .based on the range of results obtained from known reference
samples, the level of precision of the equipment or apparatus used and the
level of accuracy required for the particular application. The user should
determine the variability seen in his/her own laboratory and set in-house"
acceptance criteria that he/she judges' to be' appropriate based on. the 10c,a1
operating conditions.
Expression of Where the standard for the content of a substance described in a

Standards monograph is expressed in terms of the chemical formula for that substance
an upper limit exceeding 100% maybe stated. Such an upper limit applies
-tctheresult of the assay calculated in terms of the equivalent content of the
specified chemical formula. For example,the statement 'contains not less
than 99.0% and not more than 101.0% of C2oH24N202,HCl' implies that
the-result of the assay is not less than 99.0% and not more than 101.0%,
calculated in terms ofthe equivalent content .of C2oH24N202,HCI.
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Where the result of an assay or test is required to be calculated with

reference to. the dried, anhydrous or ignited substance, the substance free
from a specifiedsolvent .Or to the peptide content, the determination of loss
on drying, water content, loss onignition,content ofthe specified solvent .
or peptide contentis carried out by the method prescribed in the relevant
test in the monograph.
Temperature The Celsius thermometric scale is used inexpressing temperatures.
Weights and The metric system of weights and measures is employed; 51 Units have
Measures generally been adopted. Metric measures are required to have•been
graduated at 20" and all measurements involved in theanalytical operations
of the Pharmacopoeia are intended, unless otherwise stated, to be madeat
that temperature. Graduated glass apparatus-used in analyticaloperations
should comply with Class A requirements of the appropriate International
Standard issued by the International Organization for Standardization. The
abbreviationfor litre. is 'L' throughout . t.h e Pharmacopoeia.
.. . . . . .
'.' . . ...-
Atomic Weights The atomic weights adopted are the values given in the Table ofRel~tive
Atomic Weights 2001 published by the International Union ofPure and
Applied Chemistry (Appendix XXV). '
Constant Weight The term 'constant weight', used in relation to the process of drying or the
process of ignition, means that two consecutive weighings do not differ by
more than 0.5 mg, the second weighing being made after an additional
period of drying or ignition under the specified conditions appropriate to
the nature and quantity of the residue (1 hour is usually suitable).
Expression of The term 'per cent' or more usually the symbol '%' is used with one of four
Concentrations 'different meanings in the expression of concentrations according to '
Circumstances. In order that the meaning to be attached to the expression
in each instance is clear, the following notation is used: , "
Per cent w/w (% wfw) (percentage weight in weight) expresses the
number of grams of solute in 100 g of product,
Per cent wfv (% wfv) (percentage weight in volume) expresses the
number of grams of solute in 100 mL of product.
Per cent vfv (% vfv) (percentage volume in volume) expresses the
number of millilitres of solute in 100 mL of product.
Per cent vb» (% vfw) (percentage volume in weight) expresses the
number of millilitres of solute in 100 g of product.
Usually the strength of solutions of solids in liqnids is expressed as
percentage weighr in volume, of liquids in liquids as percentage volume in
volume and of gases in liquids as percentage weight in weight.
When the concentration of a solution is expressed as parts per million
(ppm), it means weight in weight, unless otherwise specified. '
When the concentration' of a solution is expressed as parts of dissolved
substance in parts of the solution, it means parts by weight (g) of a solid in
parts by volume (mL) of the final solution; or parts by volume (mL) of a
liquid in parts by volume (mL) of the final solution; or parts by weight (g)
of a gas in parts by weight (g) of the final solution.
When the concentration of a solution is expressed in molarity designated
by the symbol M preceded by a number, it denotes the number of moles of
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the stated solute contained in sufficient Purified Water (unless otherwise

stated) to produce I litre (If solution.
Water Bath The term 'water bath' means a bath of boiling water, unless water at some
other temperature is indicated. in the text. An alternative form of-heating
may be employedproviding that the required temperature is approximately
maintained but not exceeded.
Reagents The reagents required for the assays and tests ofthe Pharmacopoeiaare
.defined in appendices..The descriptions setout in the appendices. do nor
'implythat the materials. are suitable for use in medicine.
Indicators Indicators, the colours of which change over approximately the same range '
of pH, may be substituted for one another but in the event of doubt or
dispute as to me equivalence of indicators .fora particular purpose, the
indicator specified in the text is alone authoritative... '.:,
The. quantity ofanindicator solution appropriate for use in acid-base ,
titrations described in assays-or tests is if I rnl, unless otherwisestaredIn
the text,
, Any solvent required in an assay or test in which an indicator is specified
is previously neutralised to the indicator, unless a blank test is prescribed.
Caution Statements A number of materials described in the monographs and some of the
reagents specified for use in the assays and tests of the Pharmacopoeia n13Y
be injuriousto~ealth unless adequate precautions.are taken. The principles
of goodlaboratory practice and the provisions ofany appropriate
regulations such as those.issued in the United !<ingdom in .accordancewith
the Healthand.Safetr at Work etc.,Act 1974 should be observed at all times
in carrying out the assays andt~sts of the Pharmacopoeia., '.'
Attention is drawn to particular hazards in certainmonographs by means
of an italicised statement; the absence of such a statement should not
however be taken to mean .tharno hazard exists.
, '
Titles Subsidiary titles, where included, have the samesignificance as the main
titles..An.abbreviated title constructed in accordance with the directions
given in Appendix XXI A has the same significance as the main title.
Titles that are d~rived by the suitable inversion of wordsof a main or
subsidiary title, with the addition of a preposition if appropriate, are also
officialtitles. Thus, the following are all official titles: Aspirin Tablets,
Tablets of Aspirin; Atropine Injection, Injection of Atropine. ,
A title ofa formulated preparation thatincludes the full nonproprietary
name of the active ingredient or ingredients, wperethis is not included in
the title ofthe monographyis also all official title. For example, the title
Promethazine Hydrochloride Oral Solution has the same significance as
Promethazine Oral Solution and the title Brompheniramine Maleate Tablets
has the same significance as Brompheniramine Tablets.
Where the English title at the head of a monograph in the European
Pharmacopoeia is different from that at the head of the text incorporated
into the British Pharmacopoeia, an Approved Synonym has been created on
the recommendation of the British Pharmacopoeia Commission. Approved
Synonyms have the same significance as the main title and are thus official
titles. A cumulative list of such Approved' Synonyms is provided in
Appendix XXI B. '
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Where the names of pharmacopoeial substances, preparations and other

materials occur in the text they are printed with capital initial letters and
this indicates that materials ofPharmacopoeial quality must be used. Words
in the text that name a reagent or other material, a physical characteristic or
a process that is described or defined in an appendix are printed in italic
type, for example, methanol, absorbance, gas chromatography, and these imply
compliance with the requirements
. . . '.'
specified
. ,'-
in the appropriate appendix,
Chemical Formulae When the chemical composition of an official substance is known or

generally accepted.the graphic and molecular formulae, the molecular"
weight and the Chemical Abstracts Service Registry Number are normally
given at the beginning of the monograph for information. This information
refers to the chemically pure substance and is not to be regarded as an
indication of the purity of the official material. Elsewhere, in statements of
standards ofpurity and strength and in descriptions Of processes of assay, it
is evident from the context that the formulae denote the chemically pure
substances... .c, • .'....... '. •... ••.. .• .
Where.theabsolutestereochemical configuration is specified, the

. International Union, of Pure and Applied Chemistry (lUPAC) RlS and EIZ
systems of designation have been used. If the substance is an enantiomer of
unknown absolute stereochemistry the sign of the optical rotation, as
determined in the solvent and under the conditions specified in the
monograph, has. been attached to .the syste~atic name. An .indication of
sign of rotation has also been given where. this is incorporated ina trivial
name that appears onanIUPACpreferred list.
All amino acids, except glycine, have the t-configurafionunlessotherwiee
indicated. "The three-letter and one-letter symbolsused for amino acids in
peptide and protein sequences are those recommended by the Joint .
. Commission Oil Biochemical Nomenclature of the International Union of
Pure andApplied Chemistry and the International Union of Biochemistry
and Molecular Biology.
In the graphic formulae the following abbreviations are used;
Me -CH3 Bu' -CH(CH3)CH2CH3

Et -CH2Crh Bu" -CH 2CH2CH2CH3
pI -CH(CH3 ), Bu' -C(CH3 ) 3
Pr" -CH2CH2CH3 Ph -C 6H5
Bu; -CH2CH(CH3 ), Ac -COCH3
Definition Statements given under the heading Definition constitute an official

definition. of the substance, preparation Or other article that is the subject of
.themonograph.rfhey constitute instructions or requirements and are
mandatory in nature. . ..
Certain medicinal orpharmaceutical substances and other articles are
defined by reference to a particularmethod of manufacture. A statement
that a substance or article is prepared or obtained by a certain method
cons tirures part Of the official definition and implies that other methods are
not permitted. Astatement that a substance may be prepared or obtained by
a certain method, however, indicates that this is one possible method and
does not imply that other methods are proscribed.
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Additional statements concerning the definition of formulated

preparations are given in the general notice on Manufacture of Formulated
Preparations.
Production . Statements given under the headiI)g Production draw attention to particular
aspects ofJhemanufacturing process but are not necessarilycomprehensi"e.
They constitute mandatory. instructions to manlif'acrurers. They may relate,
for example, to source ma!erials! to the manufacturing process itself and its
validation and control, to in-process testing orto testing that-is to be
carried outby the manufacturer on the final product (bulk material or
. dosage form) either oriselected batche~ or on each batch prior 10 release.
These statements cannot necessarily be verified on a sample of the final
product by an independent analyst. The competent authority may establish
that the instructions have been followed, for example, by examination of
data received from the manufacturer, by inspection or by testing
appropriate samples. . • . . . . . . ". .
. Theabsence of'a section on Production does not imply that attention to
features such as those referred toab()ve is not required. A substance,
preparation or article described in a monograph-of the Pharmacopoeia is to
be manufactured in accordance with the principles of good manufacturing
practice and in accordance with relevant international agreements arid
supranational and national regulations governing medicinalproducts.
Wherein the section under the heading Production a monograph ona
vaccine defines. the charactpristics of the vaccine strain to be used, any test
·111eth0dsgi~en.for confirmingthese characteristics are provided as examples
.• ofsuitable methods, The Use of these tnethQ?sisnotmandatory.
. •Additional statements.coricerning theprcducticn of formulated
preparations are given in the general notice on Manufacture ofFormulated
Preparations.
Manufacture of Attention is drawn to the need to observeadequate hygienic precautionsin

Formulated the preparation and dispensing of pharmaceutical formulations. The
Preparations principles of good pharmaceutical manufacturing practice should be
observed.
The Definition in certain monographs for pharmaceutical preparations is
given in terms of the principal ingredients only. Any ingredient; other than
those included iri the Definition, must comply with the general notice on
Excipienrs and the product must conform with the Pharmacopoeial
requirements.
The Definition in other monographs for pharmaceutical preparations is
presented as a full formula. No deviation from the stated formula is
permitted except those allowed by the general notices on Colouring Agents
and Antimicrobial Preservatives. Where additionally directions are given
under the heading Extemporaneous Preparation these are intended for the
extemporaneous preparation of relatively small quantities for short-term
supply and use. When so prepared, no deviation from the stated directions
is permitted. If, however, such a pharmaceutical preparation is
manufactured on a larger scale with the intention that it may be stored,
deviations from the stated directions are permitted provided that the final
product meets the following criteria:
(I) compliance with all of the requirements stated in the monograph;
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(2) retention of the essential characteristics of the preparation made strictly

in accordance with the directions ofthe Pharmacopoeia.
Monographs for yet other pharmaceutical preparations include both a
Definition in terms ofthe principal ingredients and,under the side-heading
Extemporaneous Preparation, a full formula together with, in some cases,
directions for their preparation. Such full formulaeand.directions are
intended for the extemporaneous preparation of relatively small quantities
for short-term supply and use. When so prepared, no deviation from the'
stated formula and directions is permitted. If,however, such a
pharmaceutical preparation is manufactured on a larger scale with .the
intention that it may be stored, deviations from the formulaand directions
stated under the heading Extemporaneous Preparation are permitted
provided that any ingredient, other than those included in the Definition,
complies with the general notice on Excipientsand that the final product
meets the following criteria:
(1) accordance with the Definition stated in the monograph;
(2) _ compliance with all of therequireme9-tsstatedinthemonograp~;
(3) retentionofthe>essential characteristicS ofthepreparation made strictly
in accordance with the formula and directions of the Pharmacopoeia.
In the manufactureof any official preparation on a large scale with the
intention that it should be stored, in addition to following any instruction
under the headi~g Production, it is necessary to ascertain that the product
is satisfactory ""ith respect to its physical and che/ilical stability and its state
of preservation over the claitnedshelf'-life. This applies irrespective of
whethertheformula .: ofthe Pharmacopoeia .and iallY ·instructions given under
the heading Extemporaneous Preparation are foIIowedprecisely or ....
modified. Provided.that the preparation has beenshown to be stable in
other respects, deterioration due to microbial contamination may be
inhibitedbytheincorporation ofa suitable antimicrobial preservative. In
such circumstances the label states appropriate storage.conditions, the date
after which the productshould not be used and the identity and
concentration of theantimicrobial preservative.
Freshly and - The direction, given under the heading Extemporaneous Preparation, that a
Recently Prepared preparation must be freshly prepared indicates that it must be made not
more than 24 hours before iris issued for use. The direction that a . .-
preparationshould be recently preparedindicates that deterioration islikely
if the preparation is stored for longer than. about 4 weeks .at 15° to 25°.
Methods of The methods of sterilisation used in preparing .the sterile materials

Sterilisation described in the Pharmacopoeia are given in Appendix XVIII. For aqueous
preparations, steam sterilisation (heating in anautoclaye) is tile method of
choice wherever it is kn.0wn to be.suitable.Any method of sterilisationlllllst
be validated with respect to both the assuranc.e of sterility and the integrity
of the product and to. ensure that the final product complies with the
requirements of the monograph.
Water The term water used without qualification in formulae for formulated
preparations means either potable water freshly drawn direct from the
public supply and suitable for drinking or freshly boiled and cooled Purified
Water. The latter should be used if the public supply is from a local storage
tank or if the potable water is unsuitable for a particular preparation.
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. Excipients Where. an excipient for which there is a pharmacopoeial monograph is used

in preparing an official.preparation it shall comply with that monograph.
Any substance added in preparing an official preparation shall be
innocuous, shall have no adverseinfluence on the therapeutic efficacy. of the
active ingredients and shall not interfere with the assays and t~stsofihe
Pharmacopoeia. Particular care should be taken to.ensurethat such.
substances are free.fromharmful organisms: . .
Colouring Agents If in a monographfor aformulatedpreparation defined by means of a full

formula a specific colouring agent or agents is prcscribed.suitable
alternatives approvedsin the country concerned may be substituted.
Aqtimicrobial When the teITll 'suitable antimicrobial preservative' is used itis implied that
Preservatives the preparation concerned will be effectivelypreserved according to the
_ appropriate criteria applied and interpreted as described in the testfor .
efficacy ofantimicrobial preservation (Appendix XVI C). In certain
monographs for formulated preparations defined by means ofa full formula,
a specmcantimicrohial .agent or agents. may.be Prescribed; suitable ..
alternatives may be<substiruted provided that their ideiuityand
. concentration are stated on the label.
Characterlstics Statements given under the heading Characteristics are not to be

interpreted in a strict sense and are not to be regarded as official
requirements. Statements on taste are provided only in cases where this
property is a guide to the acceptability of the.material (for example, a.
material used primarily for flavouring). The status ofstateinents on
solubility is given in the general notice on Solubility. ... .- .
Solubility Statements on solubility given under the heading
Characteristics are intended as information on the approximate solubility at
a temperature between 15° and 25°, unless otherwise stated, arid are not to
be considered as official requirements.
Statements given under headings such as Solubility in ethanol express
exact requirements and ~onstirute part of the standards for the substances
under which they occur.
The following table indicates the meanings of the terms used in
statements of approximate solubilities.
Descriptive term Approximate volume of solvent

in millilitres per gram of solute
very soluble less than I
freely soluble from ·1 to 10
soluble from 10~to 30
sparingly soluble from 30 to 100
slightly soluble from. 100 to 1000
very slightly soluble from 1000 to 10 000
practically insoluble more than 10 000
The term 'partly soluble' is used to describe a mixtureof which only

some of the components dissolve.
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Identification The tests described or referred to under the heading Identification are not
necessarily sufficient to establish absolute proof of identity. They provide a
means of verifying tha t the identity of the material being examined is in
accordance with the label on the container.
Unless otherwise prescribed, identificationtests are carried out at a
temperature between IS" and 25".
Reference spectra Where a monograph refers to an infrared reference
spectrum, this spectrum is provided in a separate section of the -
Pharmacopoeia. A sample spectrum i~ considered to be concordant with a
reference spectrum if the transmission minima (absorptionmaxima) of the
principal bands in the sample correspond in position, relative intensities and
shape to those of the reference. Instrumentation software may be used to
calculate concordance with a previously recorded reference spectrum.
When tests for infrared absorption are applied to material extracted from
formulated preparations, strict concordance with.the specified reference -
spectrum may not always be possible; but nevertheless a close resemblance
between the spectrum of the extracted material.and the specified referen~e
spectrum should be achieved. _- _,_ -
Assays andTests The assays and tests-described are the official methods upon which the
standards of the Pharmacopoeia depend. The analyst is not precluded from
employing alternative methods, including methods of micro-analysis, in-any
-assay Or test if it is-known that the method used will give a result of
equivalent accuracy. Local reference materials may be used for routine
analysis, provided that these are calibrated against the official reference
materials. ill the event of doubt or dispute,_the methods of analysis; the
reference materials and the reference spectra of the Pharmacopoeia are -
alone authoritative.
Where the solvent used for a solutionis not named, the solvent is
Purified Water.
Unless otherwise prescribed, the assays and 'tests are carried out at a
temperarure between 15" and 25".
A temperature--ill a test for Loss on dryifig, where no temperature range
is given, implies a range of ±2" about the stated value.
Visual comparative tests, unless otherwise prescribed, are carried out
using identical tubes of COlourless, transparent, neutral glass with a flat
base. The volumes of liquid prescribed are for use with tubes 16 mm in
internal diameter; tubes with a larger internal diameter may beused but the
_volume of liquid examined must be increased so that the depth of liquid in
the tubes is not Jess than that obtained' when the prescribed volume of
liquid and tubes 16 mm in internal diameter are used. Equal volumes of the
liquids to be compared are examined down the vertical axis of the rubes _
against a white background or, if necessary.iagainst a.black background.
The examination is carried out in diffuse light.
Where a direction is given that an analytical operation is to be carried out
'in subdued light', precautions should be taken to avoid exposure to-direct
sunlight or other strong light. Where a direction is given that an analytical
operation is to be carried our 'protected from light', precautions should be
taken to exclude actinic light by the use of low-actinic glassware, working in
a dark room or similar procedures.
For preparations other than those of fixed strength, the quantity to be
taken for an assay or test is usually expressed in terms of the active_
ingredient. This means that_the quantity-of theactive ingredient expected to
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be present and the quantity of the preparation to be taken are calculated

from the strength stated on the label. c
In assays the approximate quantity to be-taken for examination is

indicated but the quantity actually used must not deviate bymore than
10%frorn that stated. The quantity taken is accurately weighed or
measured and the result of the assay is calculated from this exact quantity.
Reagents are measured and the procedures are carriedoutwith an accuracy
commensurate with the degree of precision implied by the standard stated
for the assay. ~ ~ ~ ~~ ~
In tests the stated quantity. to be taken for examination must be used
unless any divergence can be taken into account in conducting the test and
calculating the result. The quantity taken is accurately weighed or measured
with the degree of precision implied by the standard or, where the standard
is not stated numerically (for example, in tests for Clarity and colour of
solution), with the degree of precision implied by the number of significant
figures stated. Reagents are measured and the procedures are carried out
with an accuracy commensurate with-tills degree of precision.
The limits stated in monographs are. based on data obtained in .normal
analytical practice; they take account ofnormal analytical errors, of
acceptable variations in manufacture and of deterioration to an extent
considered acceptable. No further tolerances are to be applied to the limits
prescribed to determine whether the article being examined complies with
the requirements of the monograph. c
In determining compliance with a numerical limit, the calculated result of

a test or assay is first rounded to the number of significant figures stated,
unless otherwise prescribed. The last figure is increased by 1 when the part
rejected is equal to-or exceeds one half-unit, whereas it is not modified ~
when the part rejected -is less than a halt-unit.
~In certain tests, the concentration of impurity is given in parentheses
either as a percentage or in parts per million by weight (ppm). In
chromatographic tests such concentrations are.stared as a percentage
irrespective of the limit. In other tests they are usually stated in ppm unless
~ the limit exceeds 500 ppm. In those chromatographic tests in which a
~ secondary spot 0;' peak in a chromatogram obtained with a solution of the
substance being examined is described as corresponding to a named
impurity and is compared with a spot or peak . in a chromatogram obtained
, - -
with a reference solution of the same impurity, the percentage given in

parentheses indicates the limit for that impurity. In those chromatographic
tests in which a spot or peak In a chromatogram obtained with a solution of
the substance being examined is described in terms other than as
corresponding to a named impurity (commonly, for example, as any (other)
secondary SPOI or peak) but is comparedwith a spot or peak in a
chromatogram obtained .with a reference solution of a named impurity, the
percentage given in parentheses indicates an impurity limit expressed in
terms of a nominal concentration ofrhe named impurity. In
chromatographic tests in which a comparison is made between spots or
peaks in chromatograms obtained with solutions of different concentrations
of the substance being examined, the percentage given in parentheses
indicates an impurity limit expressed in terms of a nominal concentration of
the medicinal substance itself. In some monographs, in particular those for
certain formulated preparations, the impurity limit is expressed in terms of a
nominal concentration of the active moiety rather than of the medicinal
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substance itself. Where necessary for clarification the terms in which the
limit is expressed are stated within the monograph.
In all cases Where an impurity limit is given in parentheses, the figures
given are approximations for information. only; conformity with the
requirements is determined On the basis ofcompliance or otherwise.with
the stated test. -
The use of a proprietary designation to .identify a material used Inan
assay or test does not imply-that another equally suitable material may not
be used.
Blologlcal Assays Methods of assay described as Suggested methods are not obligatory; but.
and Tests when another method is used its precision must be not less than that .
required for the Suggested method.
For those antibiotics for which the monograph specifies a microbiological
assay the potency requirement is expressed In the monograph In
International Units_{IU) per milligram. The material is not of
pharmacopoeial quality if the upper fiducial limit of error is less than the
stated potency. For such-antibiotics the required precision of the assay is --
stated In the monograph in terms of the-fiducial limits of error about the
estimated potency.
For other substances and preparations for which the-monograph specifies_
a biological assay, unless otherwise stated, the precision of the assay is such
that the fiducial limits of error, expressed as a percentage of-the estimated
potency, are within a range not wider than that obtained by multiplying by
_a factor of 10 the square roots of the limits given In the mono-graph for the -
-- fiducialJimits.of error about the stated potency. _ -
-In allcasesftc\uciallimits of error are based on a probabilitY of 95'% -
(p= 0.95). - - -_
Where the biological assay is being used to ascertain the punty of me
material, the stated potency means the potency stated on the label In terms
of International.Units (IV) or other Units per gram, per milligram or per
millilitre. When no such statement appears on the label,-the stated potency
means the fixed or minimum potency required in the monograph. This
Interpretation of stated potency applies In all cases except where the
-monograph specifically directs otherwise. . _
Where the biological assay is being used to determine the total activity in
the container, the stated potency means the total number of International
Units (IU) or other Units stated on the label or, if no such statement
appears, the 'total activiry calculated in accordance with the instructions in
the monograph.
Wherever possible the primary standard used in an assay or test Is the
respective International Standard or Reference-Preparation established by
the World Health Organization-for international use and the biological
activity is expressed in International Units (IV). _
In other cases; where Units are referred to in an assay or test, the Unit
for a particular substance or preparation is, for the United Kingdom, the
specific biological activity contained In such an 'amount of the respective
primary standard as the appropriate International or national organisation
_Indicates. The necessary information is provided with the primary standard.
Unless otherwise directed, animals used In an assay or a test are healthy _
animals, drawn from -a uniform stock, that have not previously been treated
with any material that will Interfere with the assay-or test. Unless otherwise
stated, guinea-pigs weigh not less than 250 g or, when used in systemic
,I
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J
toxicity tests, not less than 350 g. When used in skin tests they are white or
light coloured. Unless otherwise stated, mice weigh not less than 17 g and
not more than 22 g. "
Certain of the biological assays,and tests of the Pharmacopoeia are such
that in the United Kingdom they maybecarried out only in accordance
with the Animals (Scientific Procedures) Act 1986. Instructionsincluded in
such assays and tests in the Pharmacopoeia, with respect to the handling of' , '
'animals, aretherefore confined to those concerned with the accuracy and
reproducibility of the assay or test.
Reference -Certain monographs require the use of a reference substance, a reference "
Substances and' preparation or a reference spectrum. These are chos~nwith regard to their'
Reference intended use as prescribed in the monographs of the Pharmacopoeia and
Preparations are not necessarily suitable in other circumstances.
Any information necessary for proper use of the reference substance or
reference preparation is given on the label or in the accompanying leaflet or .
brochure. Where no drying conditions are stated in the leaflet or on the
label" the substanceis to be used as received. No certificate of analysis.or
other data not relevant to, the prescribed'use of the product are provided.
, The products are guaranteed to be suitable for use for a period ofthree
months from dispatch when stored .under the appropriate conditions. The
stability of the contents of opened containers cannot be guaranteed. The
current lot is listed in the BP Laboratory website catalogue. Additional ,
information is provided inSupplemenrary Chapter I I I E . . , ,
Chemi(!~l Reference Substances The abbreviation BPCRS indicates
a Chemical Reference Substance establishedby the British Pharmacopoeia
Commission. The abbreviation CRSor EPCRSinditates aChemical '
Reference Substance established by the European Pharmacopoeia
Commission. Some Chemical Reference Substances are used for the
microbiological assay of antibiotics and their activity is stated, in
International Units, on the label or on the accompanying leaflet and' defined
in the same manner as for Biological Reference Preparations. -, ,
, Biological Reference Preparations The majority of the primary
biological reference preparations referred to are the appropriate' ,
International Standards and Reference Preparations established by the
World Health Organisation, Because these reference materials are usually
availableonly in limited quantitiesjthe European Pharmacopoeia has
established Biological Reference Preparations (indicated by the abbreviation
BRP Or EPBRP) where appropriate. Where applicable, the potency ofthe
Biological.Reference Preparations is expressed in InternationalUnits. For
some Biological Reference Preparations, where an international standard or
reference preparation does nor existythe potency is expressed in European
\ Pharmacopoeia Units.
,Storage "Statements under the side-heading Storage constitute non-mandatory

advice. The substances and prepararions described in the Pharmacopoeia
are to be stored under conditions that prevent contamination and, as far as
possible, deterioration. Unless otherwise stated in thenionograph,the
substances and preparations described in the Pharmacopoeia are kept in
well-closed containers and stored at a temperature not exceeding 25°.
Precautions that should be taken in relation to the effects of the
atmosphere, moisture, hear and light are indicated, where appropriate, in
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the monographs. Further precautions may be necessary when some

materials are stored in tropical climatesor under other severe conditions.
The expression 'protected from moisture' means that the product is to be
stored in an airtight container. Care is to be taken when the container is ..
opened in a damp atmosphere. A lowmoisrure Content may be maintained,
if necessary, by the use of a desiccant in the container provided that direct
contact with the product is avoided., ••••. .,.' '.
The expression 'protected from light' means that the product isto be
stored either in a container made of a material thaCabsorbsactiIliJ; light
sufficiently to protect the contents fromchangeinduced by such light or in
a container enclosed in an outer cover that provides such protection Or
stored in a place from which all such -Iight is excluded. '
- The expression 'tamper-evident container'. means a closed container fitted
with a device that reveals irreversibly whether the container has been
opened.
Labelling The labelling requirements of the Pharmacopoeia are not comprehensive,

and the provisions -of .rcgulations issued in accordance with the _ ,
requirements of the territory in which-the medicinal product is to be used
should be met.
, Licensed medicines intended for use within the United Kingdom must
comply with the requirements of the Human Medicines Regulations 2012,
as amended, in respecr of their labelling and packaging leaflets, together
with those regulations for the labelling of hazardous materials.
Best practice guidance on the labelling and packaging of medicines for
u~e in the United Kingdom advises thatcertain items of information are
, 'deemed critical for the safe use of the medicine (see "Best Practice
. Guidance on ,the Labelling and Packaging of Medicines" issued by the
MHRA, 2012). Further information and guidance on the labelling of
medicinal products can be found in-Supplementary Chapter I G.
Such matters as the exact form. of wording to be used and whether a-
particular item of information should appear on the primary label and
additionally, or alternatively, on the package OF exceptionallyin a leaflet are,
in general, outside the scope of the Pharmacopoeia. When the term 'label'
is used in Labelling statements or the Pharmacopoeia, decisions as to where
the particular statement should appear should therefore be made in
accordance with relevant legislation.
The label of every official formulated preparation other than those of
fixed strength also states the content of the active ingredient or ingredients
expressed in the terms required by the monograph. Where the content of
active ingredient is required to be expressed in terms other than the weight
of the official medicinal substance used in making the formulation, this is
specifically stated under the heading Labelling. Unless otherwise stated in
the monograph, the content of the active ingredient is expressed in terms of
the official medicinal substance used in making the formulation.
These requirements do not necessarily apply to unlicensed preparations
supplied in accordance with' a prescription. For requirements for unlicensed
medicines see the general monograph on Unlicensed Medicines.
Action and Use The statements given under this headingin monographs are intended only
as information on the principal pharmacological actions or ,the uses of the
materials in medicine or pharmacy. It should notbe assumed that the
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substance has no other action or use, The Statements are not intended to be
binding on prescribers or to limit their discretion,
Crude Drugs; Herbaland complementary medicines are classed as medicines under the Human
Traditional Herbal Medicines Regulations 2012, as amended. It is emphasised that, although
and Complementary requirements for the'quality of the material areprouidedin the m'!llogrdph to assist
Medicines, the registration scheme by the UK Licensing.Authority, the British Pharmacopoeia
Commission has not assessed the safety or efficacy' of the material in traditional '
use. . ,,' _
Monograph Title For traditional herbal medicines, the monograph
tide is a combination of the binomial name together with a description of
use. Monographs for the, material that has not been processed (the herbal
drug) and the processed material (the herbal drug preparation) are
published where possible. To distinguish between the two, the word
'Processed' is included in the .relevant monograph title.
Definition Under the heading Definition, the botanical name together
with any synonym is given." Where appropriate, 'for material that has not
been processed, information on the collection/harvesting and/or treatment!
drying of the whole herbal drug may begiven, For processed materials, the
method of processing, whe~e appropriate, will normally be given in a
separate section.
Characteristics References to odour are included only where this is
highly characteristic. References to taste are not included.
Control m~thods Where applicable, the control methods to be used in
monographs, are:
(a) , macroscopical and microscopical descriptions and chemical/
chromatographic tests for identification
(b) tests for absence of any related species
(c) microbial test to assure microbial quality
(d) tests for inorganic impurities and non-specific purity tests, including
extractive tests, Sulfated ash and Heavy metals, where appropriate
(e) test for Loss on drying or Water
(f) wherever possible, a-method for assaying the active constiruent(s) or
suitable marker consriruenns).
'The macroscopical characteristics include those features that can be seen
by the unaided eye or by the -use of a hand lens. When two species/
subspecies of the same plant are included in the Definition, individual
differences between the two are indicated where possible.
The description of the microscopical characteristics of the powdered drug
includes information on the dominant or the most specific characters.
Where it is considered to be an aid to identification, illustrations of the
powdered.drug may be provided.
The following aspects are controlled by the general monograph for
Herbal Drugs: they are required to be free from moulds, insects, decay;
animal matter and animal excreta. Unless otherwise prescribed the amount
of foreign matter is not more than 2% w/w. Microbial contamination should
be minimal.
In determining the content of the active constituents or the suitable
marker substances measurements are made with reference to the dried or
anhydrous herbal drug. In the tests for Acid-insoluble ash, Ash, Extractive
soluble in ethanol, Loss on drying, Sulfated ash, Water, Water-soluble ash
and Water-soluble extractive of herbal drugs, the calculations are made with
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reference to the herbal drug that has not been specifically dried unless
oth~rwise
-
prescribed
. -.-.'
ill the monograph. ,--
Homoeopathic Homoeopathic medicines are classed asmedicines under the Human Medicines
Medicines Regulations 2012, as amended. Itis emphasised that, althOJigh requirements.for
the quality of me material are prooided.in the~levant monograph-in order to . .
assist the simplified registrationscheme Qy the UKLicensing AuthlJrity, the British
Pharmacopoeia Commissionhas notassessed me safe!)! orefficacy of.the material
inuse. _ ,_ " " ..... : . . ',.'. , ." .,_ .~
All materials used for the production of homoeopathicrnedicines, .
including excipieIlts, must comply with European Pharmacopoeia or British
Pharmacopoeia monographs for those materials. Where such European
Pharmacopoeia or British pharmacopoeia monographs do not exist, each
material used for the production of homoeopathic medicines must comply
with an official national pharmacopoeia of.a Member State.
British Pharmacopoeia monographs forhomoeopathic medicines apply to
homocoparhicstocks andmother tinctures only, but may be wefacedby a
section which details the qualityreqllirem~ntsapplicable. to the. principle
component where there is no European Pharmacopoeia or British
Pharmacopoeia monograph fotthemateriaI.T\lesemonograp~salso_
include either general statements on the methods of preparation or refer to
. specific methods of preparation given in the European Pharmacopoeia.
Homoeopathicsrocksand mother tinctures undergo the further process
referred to aspotentisarionPotenrisationis a term specific to hornoeopathic
inedicinealld is a processofdillltion of stocks and mother tincruresto
produce thefinal product. •... <. .• . ...•.•. . . •. .
Identification tests are establishedfol'...the componentsin homoeopathic
stocks and usually relate to those applied to the materials USed in the
productionofthe homoeopaihicsrocks.Anassay is included for the
principal componentts) where possible. For mother tinctures, an
identification test, usually. chromatographic.is established and, where
applicable, an assay for the prin~iplecomponent(s); where appropriate,
other tests, related jothesolvent, dry matter or known adulterants, are
included.
Specifications have not been set for final homoeopathic products due. to
the high dilution used in their preparation and the subsequent difficulty in
applying analytical methodology.
Statements under Crude Drugs; Traditional Herbaland Complementary
Medicines also apply to homoeoparhlc stocks andrnothertincrures, when
appropriate.
Unlicensed The General Monograph for Unlicellsed 'Medicinesapplies to those

Medicines formulations used in human medicine.that are prepared. under a
Manufactuter's'Specials'Licence or prepared extemporaneously under the
supervision of a pharmacist, whether or notthere is a published monograph
for the specific dosage form.
An article intended for medicinal use that is described by means ofan
official title must comply with the requirements of the relevant monograph.
A formulated preparation must comply throughout its assigned shelf-life
(period of validity). The subject of any other monograph must comply
throughout its period of use.
Unlicensed medicines that are prepared under a Manufacturer's
'Specials' Licence comply with the requirements of the GeneralMonograph
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for Pharmaceutical Preparations, the.requirements of the General

Monograph for Unlicensed Medicines and, where applicable, the c •
requirements of the individual monograph for the .specific dosage form.

Unlicensed medicines prepared extemporaneously under the supervision
cofaphllrinacistcomply with therequirements of the General Monograph ..
. for Pharmaceutical Preparations, the requirements of the General.
Monograph-for Unlicensed Medicines and, where applicable,the
requirements of the individualmonograph for the specific dosage form,
While it is expected that extemporaneous preparations will demonstrate:
pharmacopoeial compliance when tested, it is recognised that it might not
be practicable to carry out the pharmacopoeial tests routinely on such
formulations ..Inthe event of doubt or dispute, the methods of analysis, the ..
reference materials and .the reference spectra of the Pharmacopoeia are
alone authoritative.
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F20 General Notices 2022
Part Ill,
,. . '
Monographs and other texts of the European Pharmacopoeiathat are incorporated

in this edition of the British Pharmacopoeia aregoverned by the general notices of
the European Pharmacopoeia; these are reprodilced below.
GENERAL NOTICES OF THE EUROPEAN
PHARMACOPOEIA
1.1. GENERAL STATEMENTS
The General Notices apply to all monographs and other texts of me
European Pharmacopoeia.
The official texts of me European- Pharmacopoeia are published in
English and French. Translations in otherlanguages may be prepared by ,
the signatory States of the European Pharmacopoeia Convention. In case of ,
doubt or dispute, the English and French versions are alone authoritative.
In the texts of the European Pharmacopoeia, the word 'Pharmacopoeia'
without qualification means the European Pharmacopoeia. The official
abbreviation Ph. Eur. may be use'd to indicate the European'
Pharmacopoeia.
The use of the tide or the subtitle of a-monograph implies that the article
complies with, the requirements of the relevant monograph. Such references
to monographs in the texts of the Pharmacopoeia are shown using' the
monograph tide and reference number in italics.
I
1
!
A preparation must comply throughout its period of validity; a distinct i
period of validity and/or specifications for opened or broached containers I
may be decided by the competent authority. The subject of any other i
monograph must comply throughout its period of use. The period of
validity that is assigned to any given article and the time from which that
period is to be-calculated are decided by the competent authority in light of
experirnental results of stability studies. I
Unless otherwise indicated in the General Notices orin the monographs, j
statements 'in monographs constitute mandatory requirements. General ,
I
chapters become mandatory when referred to in a monograph, unless such

reference is made in a way that indicates that it is not the intention to make
the text referred to mandatory but rather to cite it for information.
The active substances, excipients, pharmaceutical preparations and other
articles described in the monographs are intended for human and veterinary
use (unless explicitly restricted to one of these uses).
Quality systems The quality standards represented by monographs are valid only where the
articles in question are produced within the framework of a suitable quality
system. The quality system must assure that the articles consistently meet
the requirements of the Pharmacopoeia.
Alternative .methods The tests and assays described are the official methods llPon which the
standards of the Pharmacopoeia are based. Wim theagreement ofthe
competent authority, alternative methods ofanalysis may be used-for
control purposes, provided that the methods used enable an unequivocal
decision to be made as to whether compliance with the standards of the
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monographs would be achieved if the official methods were used, ill the
event -of doubt or dispute, the methods of analysis of the Pharmacopoeia are
alone authoritative. ~ ~
Demonstration of (1) An article is not of Pharmacopoeia quality unless it complies with all the
compliance with the ~ requirements stated in the monograph. This does not imply that
Pharmacopoeia performance of all the-tests in a ~nionograph is necessarily a prerequisite
for a manufacturer-in assessing compliance with the Pharmacopoeia
before release of a product. The manufacturer may obtain assurance
that a product is of Pharmacopoeia quality on the basis' of its 'design,
together withits control strategy and data derived, forexample, from
validation studies of the manufacturing process.
(2) An enhanced approach to quality control could utilise process analytical
technology (PAT) and/or real-time release testing (including parametric
release) strategies as alternatives to end-product testing alone. Real-time
release testing in circumstances deemed appropriate by the competent
authority is thus not precluded by the need to comply with the ~
'- Pharmacopoeia; -~ : ' , -- ~ , ~ ~ ~'
~ (3) Reduction of auimal testing: -the European Pharmacopoeia is dedicated
to phasing out the use of animals for .test purposes, in accordance with ~
the 3Rs (Replacement, Reduction, Refinement) ~set out in the European
Convention for the Protection of Vertebrate Auimals used for
'Experimentaland Other Scientific Purposes. ill demonstrating
compliance with the Pharmacopoeia -as indicated above (1),
manufacturers may consider establishing additional systems to monitor
consistency Ofproduction. \Vith the agreement uf the competent
authority, the choice of tests performed to assess compliance with the ,
Pharmacopoeia when auimal tests are prescribed is established in such a
~ way that auimal usage is minimised as much as possible.
Grade of materials Certain materials that are the subject of a pharmacopoeial monograph may
exist in.different grades Suitable for different purposes. Unless otherwise
indicated in the ~onograph, the requirements apply to all grades of the
material: In some monographs, particularly those on excipients, a list of
functionality-related characteristics that are relevant to the use of the
substance may be appended [0 the monograph for information, Test
methods for determination of one 'or more of these characteristics may be
given, also for information.
General Substances and preparations that are the subject of an individual

monographs monograph are also required to comply with relevant, applicable general
monographs. Cross-references to applicable general monographs are.not
normally given in individual monographs.
General monographs apply to all substances and preparations within the
scope of the Definition section of the general monograph, except where a
preamble limits the application, for example to substances and preparations
that are the subject of a monograph of the Pharmacopoeia.
General monographs on dosage forms apply to all preparations of the
type defined. The requirements are not necessarily comprehensive for a
given specific preparation and requirements additional to those prescribed
in the general monograph may be imposed by the competent authority.
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General monographs and individual monographs are complementary. If

the provisions of a general monograph do not apply to a particular product,
this is expressly stated in the individual
. . monograph. . .
Validation of The test methods given in monographs and general chapters have been
pharmacopoeial validated.in accordance with accepted scientific practice and current:
methods recommendations on analytical validation. Unless otherwise stated in the
monograph or general chapter, validation of the test methods by the analyst·
is not required. . .
Implementation of When implementing a pharmacopoeial method, the user must assess

pharmacopoeial whether and to what extent the suitability of the method under the actual
methods conditions of use needs to be· demonstrated- according to relevant
monographs, general chapters and quality systems.
Conventional terms. . The term 'competent authority' means the national, supranational or .
international body or organisation vested with the ·lIuthority for making -.
. decisions concerning the issue in questiori, It may; for example, be a-·
-national pharmacopoeia authority, a licensing ·authority or an official control
laboratory.· .. _
The expression 'unless otherwise justified and authorised' meansthat the
requirements have to be met, unless the competent authority authorises a
modification or an exemption where justified in a particular case.
Statements containing the word 'should' ate informative or advisory.
In certain monographs or other texts, the terms 'suitable' and
'appropriate' are used to describe 1I reagent, micro-organism, test 'method.
etc., if criteria for suitability are not described in the monograph, suitability.
is demonstrated to the satisfaction ofthe competent authority: .
.Medicinal product (a) Any substance or combination of substances
presented as having properties for treating -or preventing disease in human
beings and/or animals; or (b) any substance or.combination of substances ..
that may be used in or administered to human beings and/or animals with a
view either to restoring, correcting or modifying physiological functions by
exerting a pharmacological, immunological or metabolic action, or to
making a medical diagnosis. . . .
Herbal medicinal product Any medicinal product, exclusively
containing as active ingredients one or more herbal drugs or one or more
herbal drug preparations, or one or more such herbal drugs in combination
with one or more such herbal drug preparations.
Active substance Any substance intended to be used in the
manufacture of a medicinal product and that, when so used, becomes an
active ingredient of the medicinal product. Such substances are intended to
furnish a pharmacological activity or other direct effect in the diagnosis,
cure, mitigation, treatment or prevention of disease, or to affect the
structure and function of the body.
Excipient: (auxiliary substance), Any constituent of a medicinal
product that is not an active substance. Adjuvants, stabilisers, antimicrobial
preservatives, diluents, antioxidants, for example, are excipients.
Interchangeable Certain general chapters contain a statement that the text in question is
methods harmonised with the corresponding text of the Japanese Pharmacopoeia
and/or the United States Pharmacopeia and that these texts are
interchangeable. This impliesthat if a substance or preparation is found to
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2022 General Notices 1"23
comply with a requirement using an interchangeable. method from one of

these pharmacopoeias it complies with the requirements ofthe EUropean .
Pharmacopoeia. In the event of doubt-or dispute, the text ofthe European
Pharmacopoeia is alone authoritative,
References to Monographsand-general chapters may. contain. references to documents

regulatory issued by regulatory authorities for medicines, for example directives and.
documents notes for guidance ofthe European Union. These references are provided
for information for users for the Pharmacopoeia. Inclusion of such a . .
reference does not modify the status of the .documents referred to, which
may be mandatory or for guidance. . .
1.2. OTHER PROVISIONS APPLYING TO GENERAL

CHAPTERS AND MONOGRAPHS
Quantities In tests with numerical limits and assays, the quantity stated to be taken for
examination is approximate. The amount actually used, which may deviate
by not more than 10 per cent from that stated, is accurately weighed or
measured and the result is calculated from this exact quantity, In tests
where the limit is riot numerical, but usually depends upon comparison
with the behaviour ofa reference substance in the same conditions, the
stated quantiry is taken for examination. Reagents are used in the
prescribed amounts.
Quantities are weighed or measured with an accuracy commensurate with
the indicated degree of precision. For weighings, the precision corresponds
to plus or minus 5 units. after the last figure stated(for example, 0.25 g is to
be interpreted as 0.245 g to 0.255 g): For the measurement of volumes, if
the figure after the-decimal point is a zero or-ends in a zero (for example,
10.0 mL or .0.50 mL), the volume is measured. using a pipette, a volumetric
flask or a burette, as appropriate; otherwise, a graduated measuring cylinder
or a graduated pipette may be used. Volumes stated in microlitres are
measured using a micropipette or microsyringe.
It is recognised, however, that in certain cases the precision with which
quantities are stated does not correspond to the number of significant
figures stated in a specified numerical limit. The weighings and
measurements are then carried out with-- a sufficiently improved accuracy.
. . . -_... . . . .
J\pparatusand . Volumetric glassware complies with ClassA requirements of the appropriate

procedures International Standard issued by. the International Organisation for
Standardisation.
Unless otherwise prescribed, analytical procedures are carried out at a
temperature between IS "C and 25°C.
Unless otherwise prescribed, comparative tests are. carried out using
identical tubes of colourless, transparent, neutral glass with a flat base; the
volumes of liquid prescribed are for use with tubes having an internal
diameter of 16 mm, but tubes with a larger internal diameter may be used
provided the volume of liquid usedis adjusted (2.1.5}.Equal volumes of
the liquids to be compared are examined down the vertical axis of the tubes
against a white background, or if necessary against a black background. The
examination is carried Out in diffuse light.
Any solvent required in a test or assay in which-an indicator is to be used
is previously neutralised to the indicator, unless a blank test is prescribed.
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Water-bath The term 'water-bath' means ab~th of boiling water unlesswater at

another temperature is indicated. Other methods ofheating may be
substituted provided the temperature is near to but not higher. than 100°C
or the indicated temperature.
Drying and ignition Theterms 'dried to constant mass' and 'ignited toconstantmass' mean
to constant mass that 2cpnsecutive ",eigl:Iings do ,not differ by more thanO. 5 mg~the2nd
weighing following an additional period ofdtying or of ignition respectively
appropriate to the nature,and quantity of the residue, ,.
Where drying is prescribed using one of the expressions 'in a desiccator'
or 'in vacuo', it is carried out using the conditions.described in chapter
2.2.32. Loss on drying.
Reagents The proper conduct of the analytical procedures described in the

.Pharmacopoeia and tile reliability of theresulrsdepend.jn part, upon the
quality of the reagents used. The reagents ar~describedil1 general chapter
4. It.is .assumed that reagents of"nalyticaL~ade are used; for some
reagents, tests to determine suitabilityare included in the specifications,
_ ... c .c.- .
. __ ..-',. -.. . . . .. ~ ':- ..... . :--. _,
, Solvents Where the name of the solvent is not stated, the term 'solution" implies a
solution in water.
Where the use of water is specified or implied in the analytical
procedures described in the Pharmacopoeia or for the preparation of
reagents, water complying with the requirements of the monograph Purified
water (0008) is used, 'except that for many purposes the requirements for
bacterial endotoxins (Pur;ified waterin bulk) and microbial contamination
(Purified watedn containers) are not relevant. The term 'distilled water' "
indicates purified water prepared by distillation. .
The term 'ethanol' without qualification means anhydrous ethanol. The
... : - .. :'-:' -':- -.-
-'.. : :':'-' :"-
.. :.-:._ - .. : " -. .. .. ..
'term 'alcohol' without qualification means ethanol (96 per cent). Other
dilutions of ethanol are indicated by the term 'ethanol' or 'alcohol' followed
by a statement of the percentage by,volume of ethanol (C ZH60) required.
Expression of In defining content, the expression' 'per cent' is used according to

content circumstances with one of 2 meanings:
per cent mlm (percentage, mass in mass) expresses the number of
grams of substance in 100 g of final product;
per cent VIV (percentage, volume in volume) expresses the number of
millilitres of substance in 100 rnL of final product.
The expression 'parts per million' (or ppm) refers to mass in mass, unless
otherwise specified.
Temperature Where an analytical procedure describes temperature without a figure, the

general terms used have the following meaning:
in a deep-freeze: below -15°C;
in a refrigerator: 2 "C to 8°C;
cold or cool: 8 "C to 15°C;
room temperature: 15 "C to 25°C.
1.3. GENERAL CHAPTERS
Containers Materials used for containers are described in general chapter 3.1..General
names used for materials, particularly plastic materials, each Cover a range
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of products varyingnot only in the properties of the principal constituent

but also in the additives used. The test methods and limits for materials
depend on the formulation and are therefore applicable only for materials
whose formulation is covered by the preamble to the specification. The lise .
.of materials with different formulations, and the test methods and limits c. .
applied to them, are subject to agreement by the competent authority..
The specifications iorcontainersingeneralchapter 3,2'have been
developed for general application to containers of the stated category, but in
view of the wide variety of containers available and possible-new
• developments, the publication of a specification does not exclude the use~ in
justified circumstances, of containers that comply with other specifications,
subject to agreement by the competent authority.
Reference may be made within the monographs of the Pharmacopoeia to
the definitions and specifications for containers provided in chapter 3.2.
Containers. The genera! monographs for pharmaceutical dosage forms may,
• , under the heading Definition/Production, require the use of certain types. of
container; certain other monographs may, under the heading.Storage,"
indicate the type of container thatis recommended for use.. ' ,
1.4. MONOGRAPHS
Tides Monograph titles are in English and French in the respective versions and
there is a Latin subtitle.
,Relative Atomic The relative atomic mass (A r ) or the relative molecular mass (Mr ) is shown,
. AndMolecular as and where appropriate, at the beginning-of each monograph. The relative
Masses atomic and molecular masses and the molecular and graphic formu'lae do
not constitute analytical standards for the substances described:' '
' . '. . . ~ ,
Chemical Abstracts CAS registry numbers are included for information in monographs, where
Service (CAS)' . applicable, to provide convenient access to usefulinformation for users.
Registry Number CAS Registry Number® is a registered trademark of the American .
Chemical Society.
Definition Statements under the heading Definition constitute an official definition of

the substance, preparation or other article that is the subject of the
monograph.
Limits of content Where limits Of content are prescribed, they are
those determined by the method described under Assay.
Herbal drugs In monographs on herbal drugs, the definition indicates
whether the subject of the monograph is, for example, the whole drug Or
the drug in powdered form. Where a monograph applies to the drug in
several states, for example both to the whole drug and. the drug in
powdered form, the definition-states this,
Production Statements under the heading Production draw attention to particular

aspects of the manufacturing process but are not necessarilycomprehensive,
They constitute mandatory requirements for manufacturers, unless
otherwise stated'. They may relate, for example, to source materials; to the .
manufacturing process itself and its validation and control; to in-process
'testing; or to testing that is to be carried out by the manufacturer on the
final article, either on selected batches or on each batch prior to release,
These statements cannot necessarily be verified on a sample .of the final
article by an independent analyst. The competent authority
. _ . may" establish
.
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,-'-----C~ _
that the instructions havebeen followed,for example, by examination of

data received from the manufacturer, by inspection of manufacture or by
testing appropriate samples.' -
The absence of a Production section does not imply that attention to
'features such as those referred to above-is not required.
Choice 0/ vaccine strain, Choice a/vaccine composition The
Production section of a monograph may define the characteristics of a
, vaccine strainor vaccine composition; Unless'otherwisestated, test methods
given for verification of these characteristics are provided for information as
" examples of suitable methods. Subject to approval by the competent:
authority; other test methods may be used without validation against the
method shown in the monograph.
, Potential Due to the increasing number of fraudulent activities and cases of

Adulteration adulteration, information may be made available to Ph. Eur. users to help,
detect adulterated materials (i.e, active substances, excipients, intermediate
. products, bulk products and finished products). ,
To this purpose, a method for the detection of potential adulterants and
relev;nt limits, together with a' reminder that allstages of production and
sourcing are subjected to a suitable quality system, may be included in this
section of monographs on substances for which 'an incidenthas occurred or
that present a risk of deliberate contamination. The frequency of testing by
manufacturers or by users (e.g, manufacturers of intermediate products,
bulk products and finished products, where relevant) depends on a risk .
assessment, taking into account the level of knowledge of the whole supply
chain and national requirements." .".
This section constitutes requirements for the whole supply chain, from,
manufacturers to users (e.g. manufacturers of intermediate products, bulk
- products and finished products.where relevant). The absence of this section
does not imply that attention to features such as those referred to above is
no~ required.
- - -~; ' - - -, ,- - - '~-
.Characters 'The statements under the heading Characters are not to be interpreted 'in ~
Strict sense and are not requirements. ' .
Solubility In statements of solubility in the Characters section, the
terms used have the following significance, referred to a temperature
between 1.5 °C and 25°C.
Descriptive term , Approximate volume of solvent in millilitres

per gram of solute'
Very soluble less than I
Freely soluble from 1 to 10
- - ..-
.Soluble from 10- to 30
Sparingly soluble from 30 to 100
Slightly soluble from 100 to 1000
Very slightly soluble from " 1000 to 10000
Practically insoluble more than 10000
The term 'partly soluble' is used to describe a mixture where only some
of the components dissolve. The term 'miscible' is used to describe a liquid
that is miscible in all proportions withthe stated solvent.
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J
Identification Scope The tests givenin the Identification section are not designed to
give a full confitmation of the. chemical structure or composition of the
product; they are intended to give confirmation, With an acceptable degree
ofassurance, that the article 'conforms to the description on the label.
First and second idelltij/cations Certain monographs have
subdivisions entitled 'First identification' and 'Second identification'. The
test or. tests that.constitute the 'first identification' maybeused in all
circumstances: .The test or tests that constitute the'SecondidentificatiOI1'
may be used inpharmacies provided it can be demonstrated that the
substance or preparation isfully traceable to a batch certified to comply
with alI the other requirements ofthe monograph.
Certain monographs give two or more sets of tests for the purpose of the
first identification, which are equivalent and may be used independently.
One or more of these sets usualIy contain a.cross-reference to a test
prescribedinthe Tests section of the monograph. It maybeusedto
simplify the work of the analyst carrying out the identification and the
prescribed tests. For example, one identification setcross-refersro atestfor
enantiomericpurity while the other setgivesatestforspecillcoptical
rotationrthe intended purpose of thetwois the same, that isjverificatitlti
that the correct enantiomer is present.
Pouidered herbal drugs Monographs on herbal drugs may contain
schematic drawings of the powdered drug. These drawings complement the
description given in the relevant identification test.
, '
Tests And Ass~ys Scope The requirements arenot framed to take account of alI possible
impurities. It is not to be presumed, for example, that an 'impurity: that is
not detectable by means of the prescribedtests is tolerated if common sense
and good pharmaceutical practice require that it be absent. See also below
under Impuriti~s. ' . ,.
Calculation Where the result of a test 'or assay is required to be
calculated with reference to the dried or anhydrous substance or on some
other specified basis, the determination of loss on drying, water content or
other property is carried out by the method prescribed in the relevant test
in the monograph. The words 'dried substance' or 'anlfydro'us substance'
.etc. appear in parentheses after the result. Where a quantitative
determination of a residual solvent is carried out and a test for loss on
drying is not carried out, the content of residual solvent is taken into,'
'account for the calculation of the assay content of the substance, the
specific optical rotation and the specific absorbance. No further indication is
given in the specific monograph.
Limits The limits prescribed are based on data obtained in normal
analytical practice; they take account of normal analytical errors, of
acceptable variations in manufacture and. compoundingand of deterioration
to an extent. considered acceptable. No further tolerances are to be applied
to the limits prescribed to determine whether the article being examined
complies with the requirements of the monograph: .
In determining compliance with a numerical limit, the calculated result of
a test or assay is first rounded to the number of significant figures stated,
, unless otherwise prescribed. The limits, regardless of whether the values are
expressed as percentages or as absolute values, are considered significant to
the last digit shown (for example 140 indicates 3 significant figures). The
last figure of the result is increased by one when the part rejected is equal to
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or exceeds one half-unit, whereas it is notmodified when the part rejected

is less than a half-unit.
Indication of permitted limit ofimpurities The acceptance criteria
for related substances are expressed in monographs either in terms of
comparison of peak areas (comparative tests) or as numerical values, For
comparative tests, the approximate content of impurity tolerated, or the
sum of impurities, may be indicated in bracketsfor information only.
Acceptance or rejection is determined on the basis of compliance or non-
compliance with the stated test. If the use of a reference substance for the
named impurity is not prescribed, this content may be expressed as a
nominal concenrration of the substance used to prepare the reference
solution specified in the monograph, unless otherwise described.
Herbal Drugs For herbal drugs, the sulfated ash, total ash, water-
soluble matter, alcohol-soluble matter, \Vater content, content of essential
oil and content of active principle are calculated with reference to the drug
that has not been specially dried, unless otherwise prescribed in the
monograph, _ -
Equi'l!Cllents ~ JVhere an equivalent. is given, for the purposes of the
Pharmacopoeia only the figures ;hown are, to be' used in applying the~
requirements of the -monograph, ~ ~
Culture media The culture media described in monographs and
general chapters have been found to be satisfactory for the intended ~
purpose. However, the components of media, particularly those of ~~
biological origin, are of variable quality, and it may be necessary for optimal
performance to modulate the concentration' or some ingredients, notably:
~ peptones and meat or yeast~ exttacts,with:respect to their nutritive
properties;
buffering substances; ~ ~ ~ •
bile salts, bile extract, deoxycholate, and colouring matterj'depending ~
ori.their selective properties;
- ~antibioti~s, with resp~ect to their activity.
Storage The information and recommendations.given under the heading Storage do

no! constitute a pharmacopoeial requirement but the competent authority
may specify particular storage conditions that must be met. ~
~ The~ articles described in the Phannacopoeia are stored in such a way as ~
to prevent contamination and, as far as possible, deterioration. Where
special conditions of storage are recommended, including the type of ~
cont~iner (see section 1.3. General chapters) and limits of temperature, they
are stated in the monograph. ~ ~
The following expressions are used in monographs under Storage-with
the meaning shown. ~
an
In' airtight conta,ner Means that'the product is stored in ~aii
airtight container (3.2). Care is to be taken when the container is opened in
a ~damp atmosphere, A low moisture content may be maintained, if
necessary, by the use of a desiccant ill the container provided that direct
contact with the product is avoided.
Protected from light Means that the product is stored either in a ,
container made of a material that absorbs actinic light sufficiently to protect I
I
the contents from change induced by such light, or in a container enclosed
in an outer cover that provides such protection, or is -stored in a place from
which all such light is excluded. j
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I
J
Labelling In general, labelling of medicines _is subject to supranational and national

regulation and to international agreements, The statements under the
heading Labelling are not therefore comprehensive and, moreover, for the
purposes of the Pharmacopoeia only those statements that are necessary to
'demonstrate compliance or non-compliance .with the monograph are
mandatory. Any other labelling statements are included as ,
, recommendations. When the term 'label' is used in the Pharmacopoeia, the
labelling statements may appear on the-container" the package" a leaflet
}ccompanying the package, or a certificate of analysis accompanying the
article.as decided by the competent authority. ; _
Warnings Materials described in monographs and reagents specified for use in the
. Pharmacopoeia may be injurious to health unless adequate precautions are
taken. The principles of good quality control laboratory practice and the '
provisions, of any, appropriate regulations are to be observed at all times.
Attention is drawn to particular hazards in certain monographs by means of
a warning statement; absence of such a statement is not 'to be takeil,to
mean .that no hazard exists.'
Impurities A list of all known and potential impurities that have been shown to be
detected by the tests in a monograph may be given. See also chapter 5.10.
Control of impun'ties in substances for ph(mnaceutical use. The impurities are
designated by a letter or letters of the alphabet. Where a letter appears tobe
missing, the impurity designated by this letter 'has been deleted from the list
during monograph development prior to publication or during monograph
revision.
Funcrionallty> Monographs on excipients may have a' section on fuiictionality-related '

related characteristics: The characteristics;' any test methods for determination and
-Characteristics of any tolerances are not mandatory reguirements; they may nevertheless be,
ExCipients relevant for use .of the excipient and are given for information (see also
section 1.1. General statements). -
Reference Certain monographs require the use of reference standards (chemical

Standards reference substances, herbal reference standards, biological reference
preparations, reference spectra). See also chapter 5.12. Reference standards.
The-European Pharmacopoeia Commission establishes the official reference
standards, which' are alone authoritative in case of arbitration. These
reference standards are available from the European Directorate for the'
Quality of Medicines & HealtlrCare (EDQM). Information on the available
reference standards and a batch validity statement can be obtained via the
EDQM website.
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1.5. ABBREVIATIONS AND SYMBOLS
A Absorbance mp Melting point

~~
Specificabsorbance Refractive Index
A, Relative atomic mass Ph. Eur. U. European PhannacopoefaUnlr
l~]~ Specificoptical rotation _ ppb _Parts per billion (micrograms per kilogram)_
bp Boiling point ppm Pans per :million (milligrams per kilogiam)
BRP Biological reference preparation R Substance or solution defined under 4. R~gents _
CRS Chemical referencesubstance RF Retardation factor (see chapter 2.2.46)
dJ8 Relativedensity R" - Used m cltromatography to" indka~ the ratio of
A Waveleflgth the distance travelled by Ii substance (0 the
distance trayelled by-a reference'substance
HRS Herbal reference standard
RV Substance used as a primary.standard in
IU International VOir volumetric analysis (chapter 4.,2.1)
M~
Molarity
M - '
r Relativemolecular mass
-Abbreviations used in- the

-
monographs, on immunoglobulins,
-~- -. ~
hnmunosera and vaccines
CFU Colony-fanning .Urii~' LotIO dose The largest quantity oj aroxln that, in the"
ill50 The statistically determined.qUantity..of a condirionscf.rhe resr, when-mixed with 0.1 IU
substance that, when administered by the .Qf antitoXin and edminisrered by_the'specified
specifiedroute, may be expected to 'cause the rcute.-does not cause symptoms'oftoXicity in
death of 50 per cent of me test animals within . the test animals within a given period
l! given period ." Lfdcse The, quantityof toxin or toxoid thatflocculates
MID "Minimum lethal dose in the shortest time with 1'ill of antitoxin
L+/IO dose The smallesr ,qu_~nti[y ofa to·xin that, in 'the CCID" The statistiCally determined quantity of virus
-"conditions oflhe',test',W.ben miXed,wi.m'a1 IU thai may,~e:eXpeCted-to in(eet 50 'per, cent of
of anti~,()xin and administered by tile,specified the.-ceIl·OiItures'to'which it'is added ,-
route, "causes the death Qf the test animalS Th~ st;iisuciri~.~etetmined 'quan!:i,ty :~f virus
within_~ given period __' ' _that may be'expected .[0 ipfect .50 per cent of
'L+ dose. The__ s~~Uest'~WtrititY--9f~ toxin,th~t,.in the the, fe~sed e~',into. ",IDch it is in,otulated
conditions,of the .test, 'when mixed with 1 ill of ID,. The, sta~sPtalIY,determined quantitY, ofa. ~ .
antitoxin-and adIDiniste~d by" the specified- that maY:,be expectedto infect 50 per 'Cent of
rome, 'causes-the de~\:h 'of the test·animals :'theanittuHtdmo ;whIch it is.iJ?octl1ated '
within a given. period The 'statistically .de(.~ed dose of a vaccine
IdIOO dose The'!;IJ!lalle$t quantitY of a to;dn min, in the.', that,.in the conditioflS of the test" tnay. be
conditions ofthe-fest~-when-mixed,with 0.01 ,.- -,,::._:_expect~cUP protec;;t5P per cent of the aDimals.
1U of aptiloxin and,inlected intiacuraneously a~inn a cb,aUenie·~dose ofthe.'micro::Qrganis~s
causes a characteristic reaction af the site Q.f or toxins ~gainSt which,it is active
injecriori within a.gfven period - nie 'statiStically, dei~ed dOse of ~ vaccine' -
Lp/IO dose The smallest quantitY of toxin ·th~[,' in the - - that, in ·the, condinons of the.test, may be'
conditions of the test, when nuxed with 0.1 IV expectedto induce ,specific antibodies in 50 per
of antitoxin and administered by the'specified cent of me animals for the relevant vaccine
route, causes parelysisin the test annnaJs withiri ~tigens _ _,
a given period - PFU Pock-fumiing Units'or plaque-fonning units
SPF Specified-pathogen-fre<
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J
Collections of micro-organisms
ATCC American Type Culture Collection, NGrC National Collection of Type C~tUres
,10801 University Boulevard _ . Central Public Health Laboratory
Mana';." Virginia 201l0-2209, USA: . Colindale Avenue
C.I.P. Collection de, Bacteries de l'Iflstitut.Pasteur London NW9 5HT, Great Britain
B.P..52, 2~=rue du Docreur Roux " . NCYC National Collection of Yeast" Cultures
-75724-Paris Cedex 15, Fiance AFRC Food Research Institute
Jl'<U International MycoJogicallnstitute Cotney Lane
, Bakeham Lane Norwich NR4 7UAJ Great Britain
Surrey TW20 9TI, Great Brita'in ." NITE Biological Resource Center
J.P. Collection Nationale de Culture de Department of Biotechnology
Microorganiames (C:N.C.M.)-: National Institute' of Technology and
Institut Pasteur Evaluation
25) rue du Docreur Roux 2~5-8 Kazusakamatari,Kisarazu-shi, Chiba,
75724 Paris Cedex 15, Fnmce 292-0818
NClMB National Collectionof Industrial and .Marine Japan . .
Bacteria Ltd , - - - .5.5.1. SrarensSenun Iusrlnu _

23 St Macher Drive 80 Amager' Bo~lektd~ -Cop~nh~gen, Denmark ,
Aberdeen AB2,lRY, Great Britam
" ,'-
NCPF National Collection of Pathogenic F\U!&i
London School of Hygiene and Tropical
Medicine
Keppel Street
London WClE 7Hr, Great Britain
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1-32 .General Notices 2022
1.6. UNITS OF THE INTERNATIONAL SYSTEM (SI) USED IN .

THE PHARMAcOPOEIA AND EQUIVALENCE WITH .
OTHER UNITS .
International The International System of Units comprises 2 main classes of units,
System Of Units namely base units and derived units' : The base units are the metre, the
(SI) kilogram; the second, the ampere, the kelvin, the mole and the candela,
The derived units are formed as products of powers of the base units.
'.according to the algebraic relationships linking the corresponding quantities.
Some of these derived units have special names and symbols. The
derived units used in the Pharmacopoeia are shown in Table 1.6.-1.
Some important and widely used units "outside the International System
'are-shown in Table 1.6.-2.
The prefixes shown in Table 1.6.-3 are used to form the names and
symbols of the decimal multiples and submultiples of SI units.
Table 16 -1 "- Derivedunits usedin the European

- Pharmacopoeia andequioalence with other units
I·
- . .
-
-. Quantity - Unit . - .
. -
Conve:t's~on of other unitS-into Sf
Name- Symbol Name - Symbol Expression Expres~ori in -
- units--
-In-Sf base other SI units
units -
Wave·number v one.per metre 11m : m-' - -

.
Wavel~ngth !- micromerre pm lO'-6m

- .
.
- .
•• narioJriette om' lO"1m .
.. . .
- . .
• 2
Ate, A,S square meti:e m2 .
-'_ID - .
.
- , 6
Vol~c:;-':'---- V - cubic metre m3 =- m' ImL=l= =10- m'
.
_,¥requ.eDCy I' . . hertz Hz ,-I .
.. -
. . ' .
Density I p . '·'I;il'!gram pet kglm' kg·m'--3 1 glmL = I glom' = 10' kg·m-~

I
. I cubicmetre
Velocity;, speed I
'II me~pef--- mI, m·s-1 -
.
..
second I
- -0
.
Force F newton - N m.kg_g-2 1. dyne = 1 g-em-s-2 = 1-0-5 N

I -
I kp = 9.806 65 N
.
Pressure, stress p pascal Pa m:"' 1.Iqi.s-2 . N ·m'-2
,
l,dyne/em ':;;: 10-;:-1_Pa:;;:, 10-1 N·m-~
I 'lID = 101 325 Pa =-101.325 Id',

, 1 bar = 10' Po = 0.1 MP,
I I mm Hg = 133322 387 Pa
- . I Torr = 133.322 368.Pa
.
~
.
- , . [ - I psi = 6.894 757 Id', , ,
.
. Dynamic n pascal second Pa·s m-1·kg·s-l N·s·m-2 1 P = 10- I'pa·s:;;: 10-1 N·s·in-~
"viscosity . 1 cP = 1 ml'a-s
' "':'J - - I';"": -
Kinematic. v square metre m% m2·s-1 Pa.s.m "kg-. 1 St =-1 cm 2·s-J = JO-1 m 2·s-1
viscosity - pee.second N-m-s.kg-' .
1 The dqItlitions o/the units usedin the InternatitmoT S,YS'tetn aregiven in me' booklet 'I.e Syste'me .Imernaiional d'Uniris, (SIr, publiihed bj l/uJ .
Buiroa Ihtemotional des Poids ei M.esures"PqviUon de BreuYiJ, F-92310 Seures. -
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Quantity Unit ,
I Symbol
Converslri~ of other units into sr
Name Symbol "Name Expression Expression' in
units
inSI base other SI units
I I units
Energy ,
W ioule: J -m2 ,"k'g-s-2> , N·m '-I'erK;, '1 cm2 ,g og"""2 == 1', dyne-em =
IO-~ J ' ,
" 'J cal = 4,1868J
I
Power, P waH ,
W m2·kg:,s-J" ' N·mos-I, l-eIWS =) ,dyne-cJ¥.s-t ,=
1~,-1 W = lO~7 N'~'S-I = 10--:1 JS~,1
- '-I
radiant flux Js
Absorbed dose D gray Gy m2·s-2 , J-kg-' I red = 10-2 Gy

(of radiant
energy)
Electric U 'volt V m 2. ,l{g.s-3·A- 1 W,A-'

I
potentia)
difference, -
voftage I
" - ,
'Electric R ohm II 2 'kg '-) A-2

m,' , ,S ,', V.A~j
,
resistgace ,
- -,\f.' -
Electric';~b<lrge
-'
C ,
, I
Q coulomb As
Activity . A becquerel Bq s-' I Ci = 37·10" Bq = 37·10' s-'

referred to a
radionuclide I I
, ,
. . , .
-
Concentration' c .: mole per -moUrn), I mol·m-3 lll\ol!L = 1 M = I mol/elm' ='10'_
(ofamountof .:'cubicmetre
I mQl'm~3, -
,
,
subStance), ,
,
I '
molar '
cencentraticn ,
I ,Mass P ' 'kil~~ per _ kglm' , kg-m-, I gIL = I gldm' = I kg,m-' ,
concentration cubic metre' .

,
Catatync Z ketal kat mol-e?' I '

activity
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Table 1.6.-2, -Non-81 units accepted for use with the 81 units
Quantity Uni~ Value In Sl units
Name, SJ'Dlbol
Time minute min 1 min ==- 60 s

,-
,
hour h I h = 60 min = 3600 s

,
day d Id~24h=86400"
PI~e angle degree 0

10 = (rt! f80) rad
Volume litre L 1 L == 1 drrr' == 10- 3 m3 I
Mass tonne t 1 [= 10' kg,
dalton Da 1 Da = 1.660539040(20) x 10-27 kg
Rotational revolution. rlmin - ' 1 rlmin = (1/60) ,-I

-
frequency per minute -
- '
Energy , electronvolt _ eV leV=1.602176634 x 10-19J

"
Table 1 6 -3 - Decimal multiples and sub~multiples of $1 units

Factor Prefix SJ'Dlbol Factor Prefix SJ'Dlbol
10 1,8 _exa E ,
10- 1 deci d
,
-
'10 15 - p 10-2
- peta : ,
" '- centi c

12
• -
10 (era T - 10-' -milli ,
,m "
-
-10' I giga G 10- 6 micro ~
,
,
10· mega M 1- 10-9, neno , n
I
'0'
,I Idlo k
I 10- 12 pica I p
-
102 hecto b 1O-1~ femto f
I -~101 deca da 10- 16 atto i a

-
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Notes 1. In the Pharmacopoeia, the Celsius temperature.is used (symbol t). This is
defined by the following equation:
t = T- To
where 1'0 = 273.15 K by definition. The Celsius or centigrade
temperature is expressed in degrees Cclsius.Isymbol "C), The unit
'degree Celsius' is equal to the unit 'kelvin', " ' "
2. The practical expressions of concentrations used in the Pharmacopoeia are
defined in the General Norices.c".". ' , ',' _ -
3. The radian is the plane 'angle between two radii of a circle thatcur off on
the circumference an arc equal in.length to the radius.
4. In the Pharmacopoeia, conditions of centrifugation are defined by
reference to the acceleration due to gravity (g):
5. Certain quantities without dimensions are used in the Pharmacopoeia:

relative density (2.2.5), absorbance (2.2.25), specific absorbance (2.2.25) and
refractive index (2.2.6).
6. The microkatal is defined as the 'enzymic activity that, under defined
conditions, produces the transformation (e.g, hydrolysis) of I micromole of
the substrate per second. - --
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-
, ,
----'-----~---_. ---' - - ,
-'.'--'
I
II
I
I
i
·I
1
1
J
I
J
·i,
·
·j
':1
.i
·!
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Monographs
Medicinal and Pharmaceutical

Substances (A to I)
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j
2022 General Monographs 1-39
PRODUCTION
MEDICINAL AND PHARMACEUTICAL Substances for pharmaceutical use are manufactured by
SUBSTANCES procedures that are designed to ensure a consistent quality
and comply with the requirements of the individual
monograph or approved specification.
The manufacture of active substances must take place under
Substances for Pharmaceutical conditions of good manufacturing practice.
The provisions of general chapter 5.10 apply to the control of
Use impurities in substances for pharmaceutical use"
(ph. Bur. monograph 1034) Whether or not it is specifically stated in the individual
PO." _ monograph that the substance for pharmaceutical use:
- is a recombinant protein or anomer substance obtained as
DEFINITION
a direct gene product based on genetic modification,
Substances for pharmaceutical use are any organic or
where applicable, the substance also complies with the
inorganic substances that are used as active substances or
requirements of the general monograph Products of
excipients for the production of medicinal products for
recombinant DNA ,ochnology (0784);
human or veterinary use. They may be obtained from natural
- is obtained from animals susceptible to transmissible
sources or produced by extraction from raw materials,
spongiform encephalopathies other than by experimental
fermentation or synthesis. challenge, where applicable, the substance also complies
This general monograph does not apply to herbal drugs, with the requirements of the general monograph Products
herbal drugs for homoeopathic preparations, herbal drug with risk of transmuting agents of animal spungifonn
preparations, herbal drug extracts, or mother tinctures for encephalopathies (1483);
homoeopathic preparations, which are me subject of separate - is a substance derived from a fermentation process,
general monographs (Hetbal drugs (1433), Herbaldrugs for whether or not the micro-organisms involved are modified
homoeopathic preparations (2045), Herbaldrug by traditional procedures or recombinant DNA (rDNA)
preparations (1434), Herbal drug extracts (0765), Mother technology, where applicable, the substance also complies
tinctures for homoeopathic preparations (1019). It does not with the requirements of the general monograph Produas
apply to raw materials for homoeopathic preparations, except of'fermentation (1468).
where there is an individual monograph for the substance in
If solvents are used during production, they are of suitable
the non-homoeopathic pan of the Pharmacopoeia.
quality. In addition, their toxicity and their residual level are
This monograph does not apply to chemical precursors for taken into consideration (5.4). If water is used during
radiopharmaceutical preparations which are the subject of a production, it is of suitable quality.
separate monograph (Chemiallprecursors for
The identity of elemental impurities derived from
radiopharmaceutical preparations (1902). intentionally added catalysts and reagents is known, and
Where a substance for pharmaceutical use not described in strategies for controlling them should be established using the
an individual monograph of the Phannacopoeia is used in a principles of risk management.
medicinal product prepared for the special needs of
If substances are produced or processed to yield a certain
individual patients, the need for compliance with the present
form or grade, that specific fonn or grade of the substance
general monograph is decided in the light of a risk complies with the requirements of the monograph. Certain
assessment that takes account of the available quality of the
functionality-related tests may be described to control
substance and its intended use. properties that may influence the suitability of the substance
Where medicinal products are manufactured using and subsequently the properties of dosage forms prepared
substances for pharmaceutical use of human or animal origin, from it.
the requirements of chapter 5.1.7. Viral safay apply. Powdered substances May be processed to obtain a certain
Substances for pharmaceutical use may be used as such or as degree of fineness (1.9.35).
starting materials for subsequent formulation to prepare
Compacted substances Are processed to increase the particle
medicinal products. Depending on the formulation, certain
size or to obtain particles of a specific form and/or to obtain
substances may be used either as active substances or as
a substance-with a higher bulk density.
excjpients. Solid substances may be compacted, coated,
granulated, powdered to a certain fineness, or processed in
Coated aaioe substances Consist of particles of the active
other ways. A monograph is applicable to a substance substance coated with one or more suitable excipients.
processed with an excipient only where such processing is Granulated active substances Are particles of a specified size
mentioned in the definition section of the monograph. and/or fonn produced from the active substance by
granulation directly or with one or more suitable excipients.
Substance for pharmaceutical use of spe<ial grade Unless
otherwise indicated or restricted in the individual If substances are processed with exclpients, these excipients
monographs, a substance for pharmaceutical use is intended comply with the requirements of the relevant monograph or,
for human and veterinary use, and is of appropriate quality . where no such monograph exists, the approved specification.
for the manufacture of all dosage forms In which it can be Where active substances have been processed with excipienrs
used. to produce, for example, coated or granulated substances, the
Polymorphism Individual monographs do not usually specify processing is carried out under conditions of good
crystalline or amorphous forms, unless bioavailability is manufacturing practice and the processed substances are
affected. AU forms of a substance for pharmaceutical use regarded as intermediates in the manufacture of a medicinal
comply with the requirements of me monograph, unless product.
otherwise indicated.
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1-40 General Monographs 2022
CHARACTERS Table 2034.-2. - Reporting, identification and qualification of

The statements under the heading Characters organic impurities in peptides obtained by chemical synthesis
(e.g. statements about the solubility or a decomposition Reporting Identification Qualification
point) are not to be interpreted in a strict sense and are not threshold threshold threshold
requirements. They are given for information. > 0, I per cent > 05 per cent > 1.0 per cent
Where a substance may show polymorphism, this may be
stated under Characters in order to draw this to the attention Specific thresholds may be applied for impurities known to
of me user who may have to take this characteristic into be unusually potent or to produce toxic or unexpected
consideration during formulation of a preparation. pharmacological effects.
IDENTIFICATION For DNA reactive impurities) the requirements of ICH
Where under Identification an individual monograph Guideline M7 Assessment and Control of DNA Reactive
contains subdivisions entitled 'First identification' and (Muragem'c) lmpun',ies in Pharmaceuticals ro Limit Potential
'Second identification" the test or tests that constitute the Carcinogenic Risk must be complied with for active substances
'First identification' may be used in all circumstances. to be used in medicinal products for human use) in cases
The test or tests that constitute the 'Second identification I defined in the scope of the guideline.
may be used in pharmacies only, provided it can be If the individual monograph does not provide suitable control
demonstrated that the substance or preparation is fully for 'a new impurity) a suitable test for control must be
traceable to a batch certified to comply with all the other developed and included in the specification for the substance.
requirements of the monograph. The implementation of the The requirements above do not apply to biological and
tests under the second identification is subject to national biotechnological products, oligonucleotides) products of
regulation. fermentation and semi-synthetic products derived therefrom,
Certain monographs give two or more sets of tests for the to crude products of animal or plant origin or herbal
purpose of the first identification, which are equivalent and products.
may be used independently, One or more of these sets Elemental Impurities
usually contain a cross-reference to a test prescribed in the Permitted daily exposures for elemental impurities
Tests section of the monograph. It may be used to simplify (e.g. as included in the ICH Q3D guideline, the principles of
the work of the analyst carrying out the identification and the which are reproduced in general chapter 5.20. Elemental
prescribed tests. For example, one identification set cross- impuniies) apply to the medicinal product. Individual
refers to a test for enantiomeric purity while the other set monographs on substances for pharmaceutical use therefore
gives a test for specific optical rotation: the intended purpose do not contain specifications for elemental impurities unless
of the two is the same, that is, verification that the correct otherwise prescribed.
enantiomer is present.
Residual solvents
TESTS Are limited according to the principles defined in chapter
Polymorphism (5.9) 5.4) using general method 2.4.24 or another suitable method.
If the nature of a crystalline or amorphous form imposes Where a quantitative determination of a residual solvent is
restrictions on its use in preparations, the nature of the carried out and a test for loss on drying is not carried out)
specific crystalline or amorphous fonn is identified, its the content of residual solvent is taken into account for
morphology is adequately controlled and its identity is stated calculation of the assay content of the substance) the specific
on the label. optical rotation and the specific absorbance.
Related substances Microbiological quality
Unless otherwise prescribed or justified and authorised, Individual monographs give acceptance criteria for
organic impurities in active substances are to be reported) microbiological quality wherever such control is necessary.
identified wherever possible) and qualified as indicated in Table 5.1.4.-2. - Acceptance criteria for microbiological quality of
Table 2034.-1 or in Table 2034.-2 for peptides obtained by non-sterile subsumces for phosmaceuiicol use in chapter 5.1.4.
chemical synthesis. Microbiological quality of non-nenle pharmaceutical preparations
and substances forpharmaceutical use gives recommendations
Table 2034.-1. - Reporting, identification and qualiji<a,ian of on microbiological quality that are of general relevance for
organ~ impurities in aaive substances substances subject to microbial contamination. Depending on
Use Maximum Reporting ldentificatlon Quallflcation the nature of the substance and its intended use) different
dBlly threshold threshold threlIhold
acceptance criteria may be justified.
d...
Human use :5 2 yJday > 0.05 per > 0.10 per cent > 0.15 per cent Sterility (2.6.1)
orbuman cent or a daily intake or a daily intake If intended for use in the manufacture of sterile dosage forms
and of> 1.0 mg of> 1.0mg without a further appropriate sterilisation procedure) or if
veterinary (whichever ls (whicheveris offered as sterile grade) the substance for phannaceutical use
u"' the lower) the lower)
complies with the test for sterility.
Hwnan use > 2 glday > 0.03 per > 0.05 per cent > 0.05 per cent
or human cent Bacterial endotoxin. (2.6.1<f)
and The substance for pharmaceutical use complies with the test
veterinary for bacterial endotoxins if it is labelled as a bacterial
u"' endotoxin-free grade or if it is intended for use in the
Veterinary No. > 0.10 per > 0.20 per cent > 050 per cent
use only applicable cent manufacture of parenteral preparations or preparations for
irrigation without a further appropriate procedure for the
removal of bacterial endotoxins. The limit, when not
indicated in the individual monograph) is determined in
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2022 Abacavir Sulfate 1-41
accordance with the recommendations of general chapter

Abacavir Sulfate ****
5.1.10. Guidelines for using the test/or bacterial endosoxim, •* •
Pyrogens (2.6.8) (Ph. Eur. monograph 2589) *****
If the test for pyrogens is justified rather than the test for
bacterial endotoxins and if a pyrogen-free grade is offered,
the substance for pharmaceutical use complies with the test
for pyrogens. The limit and test method are stated in the
individual monograph or approved by me competent
authority. Based on appropriate test validation for bacterial
endotoxins and pyrogens, the test for bacterial endotoxins
may replace the test for pyrogens.
Additional properties
Control of additional properties (e.g. physical characteristics,
functionality-related characteristics) may be necessary for 671 188062-Sa-2
individual manufacturing processes or formulations. Grades
(such as sterile, endotoxin-free, pyrogen-free) may be Action and use
produced with a view to manufacture of preparations for Nucleoside reverse transcriptase inhibitor; antiviral (HIV).
parenteral administration or other dosage forms and Preparations
appropriate requirements may be specified in an individual Abacavir Oral Solution
monograph. Abacavir Tablets
ASSAY Abacavir, Zidovudine and Lamivudine Tablets
Unless justified and authorised, contents of substances for Abacavir and Lamivudine Tablets
pharmaceutical use are determined, Suitable methods are
P1>E" _
used.
LABELLING DEFINITION
In general, labelling is subject to supranational and national Bis[[(IS,4R)-4-[2-amino-6-(cyclopropylamino)-9H-purin-9-
regulation and to international agreements. The statements yl]cyclopent-2-enyl]methanol] sulfate.
under the heading Labelling therefore are not comprehensive Content
and, moreover, for the purposes of me Pharmacopoeia only 99.0 per cent to 101.0 per cent (anhydrous substance).
those statements that are necessary to demonstrate
CHARACTERS
compliance or non-compliance with the monograph are
Appearance
mandatory. Any other labelling statements are included as
White or almost white powder.
recommendations. When the term 'label' is used in the
Pharmacopoeia, the labelling statements may appear on the Solubillty
container, the package, a leaflet accompanying the package or Soluble in water, practically insoluble in ethanol
a certificate of analysis accompanying the article, as decided (96 per cent) and in methylene cbloride.
by the competent authority. IDENTIFICATION
Where appropriate, the label states that the substance is: A. Infrared absorption spectrophotometry (2.2.24).
- intended for a specific use; Comparison abaca"ir sulfale CRS.
- of a distinct crystalline form;
B. Enantiomeric purity (see Tests).
- of a specific degree of fineness;
- compacted; C. Solution S (see Tests) gives reaction (a) of sulfates
- coated; (2.3.1).
- granulated; TESTS
- sterile; Solution S
- free from bacterial endotoxins; Dissolve 0.250 g in waterR and dilute to 25.0 mL with the
- free from pyrogens; same solvent.
- containing gliding agents.
Bnendomcric purity
Where applicable, the label states: Liquid chromatography (2.2.29).
- the degree of hydration;
Solution A Mix 0.5 mL of t,;fiuuroacetic acidR and 100 mL
- the name and concentration of any excipient.
of methanol R.
_________________ ~ P1>E"
Solution B J\.lix 30 volumes of methanol R J 30 volumes of
2-propanol Rand 40 volumes of heptane R.
Test solution Dissolve 40 mg of the substance to be
examined in 30 mL of solution A. Sonicate until dissolution
is complete. Add 30 mL of 2-propa"ol R and dilute to
100.0 mL with heptane R.
Reference solution (a) Dissolve 2 mg of abacavir for system
suitabJ,iy CRS (containing impurities A and D) in 1.5 mL of
solution A. Sonicate until dissolution is complete.
Add 1.5 mL of 2-propanol R and dilute to 5.0 mL with
heptane R.
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1-42 Abacavir Sulfate 2022
Reference solution (b) Dilute 1.0 mL of the test solution to Time frlobUe phase A hlobUe phase B
100.0 mL with solution B. Dilute 1.0 mL of this solution to (min) (per cent I'll? (per cent PlY)
10.0 mL with solution B. 0-5 95 5

5 - 25 95 ---> 70 5 ---> 30
Column:
25 - 40 70 10 30 90
- size: 1= 0.25 rn, 0 = 4.6 mm; ---> ->
- stationary phase: amylose den'van'tie of silica gelfor chiral

separation R (10 ~); Flow rare 1.0 mUmin.
- temperature: 30 "C. Detection Spectrophotometer at 254 nm.
Mobile phase: Injection 20 tll-.
- mobik phase A: diethylamine R, 2-propanol R, heprane R Identification of impurities Use the chromatogram supplied
(0.1: 15:85 VIVIV); with abacavir for peak idenri/ication CRS and the
- mobile phase B: heprane R, 2-propanol R (50:50 VIV); chromatogram obtained with reference solution (a) to
identify the peaks due to impurities B and D.
Tim' Mobile phase A Mobile phase B
(min) (pel' cent VIV) (per cent YJ1I) Relative retention Widl reference to abacavir (retention
time = about 22 min): impurity D = about 1.04;
0-25 100 0
impurity B = about 1.3.
25 - 27 100 ..... 0 0 ..... 100
27 - 37 0 100 System suitab,1ity Reference solution(a):
- peak-w-1Jalley ratio: minimum 3.0 J where Hp == height
above the baseline of the peak due to impurity D and
Flow rare 1.0 mUmin.
H" == height abovethe baseline of the lowest point of the
Detection Spectrophotometer at 286 nrn. cUIVe separating this peakfrom the peakdue to abacavir.
Injection 20 ut, Limits:
Identification of impurities Use the chromatogram supplied - impurity B: not more than twice the area of the principal
with abacavir for sys<em suirabiliry CRS and the chromatogram peak in the chromatogram obtained withreference
obtained with reference solution (a) to identify the peaks due solution (b) (0.2 per cent);
to impurities A and D. - unspecified impurities: for each impurity, not more than the
Relau've retention With reference to abacavir (retention area of the principal peak in the chromatogram obtained
time = about 17 min): impurity D = about 0.8; withreference solution (b) (0.10 per cent);
=
impurity A about 0.9. - total: not more than 5 times the area of the principal peak
System suitability Reference solution (a): in the chromatogram obtainedwithreference solution (b)
- resolution: minimum 1.5 between the peaks due to (0.5 per cent);
impurities D and Aj minimum 1.5 between the peaks due - disregard limit: 0.5 times the area of the principal peak in
to impurity A and abacavir. the chromatogram obtained withreference solution (b)
(0.05 pet cent).
Limit:
- impurity A: not more than 3 times the area of the Water (2.5.32)
principal peakin the chromatogram obtained with Maximum 0.5 per cent, determined on 60.0 mg.
reference solution (b) (0.3 per cent). Sulfated ash (2.4.14)
Related substances Maximum 0.2 per cent, determined on 1.0 g.
Liquid chromatography (2.2.29). Prepare the solutions ASSAY
immediately be/ore use and transfer them to lcw-adsorption, inert Dissolve 0.300 gin 50 mL of waterR. Titrate with 0.1 M
glass vials. sodium hydroxide) determining the end-point
Test solution Dissolve 25 mg of the substance to be potentiometrically (2.2.20).
examined in water R and dilute to 100.0 mL with the same 1 mL of 0.1 M sodium hydroxide is equivalent to 33.54 rng of
solvent. Sonicateuntil dissolution is complete. C,.H.aN.,O,S.
Reference solution (a) Dissolve 2.5 mg of abacavir for peah
IMPURITIES
identijicauim CRS (containing impurities B and D) in
Specijied impurities A, B.
10.0 mL of water R.
Otherdete<rable impurities (the following substances would, if
Reference solution (b) Dilute 1.0 mL of the test solution to
present at a sufficient level, be detected by oneor other of the tests
100.0 mL with waterR. Dilute 1.0 mL of this solution to
in the monograph. They are limited by the general acceptance
10.0 mL with waterR.
criterion for otnerlunspedfied impurities andlor by thegeneral
Column: monograph Subsrances for pharmaceutical use (2034). It is
- size: 1= 0.15 m, 0 = 3.9 mrn; therefore not necessary to identify these impurities for
- stationary phase: end-capped octade<y/silyl silica gelfor demonstration of compliance. See also 5.10. Control of impurities
chromarography R (5 urn); in substances for pharmaceutical we) C, D, EJ F.
- <emperalUre: 30°C.
Mobile phase:
- mobile phase A: dilute 0.5 mL of lrifluoroacetic acidR in
1000 mL of waterR;
- mobile phase B: waterR, methanol R (15:85 VIV);
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2022 Acacia 1-43
A. [(IR,4S)-4-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl]
cydopent-2-enyl]methanol, F. 6-(cyclopropylamino)-9-[( I R,4S)-4-[[(I, J-dimethylethyl)
oxy]methyl]cyclopent-2-enyl]-9H-purine-2-amine.
_________ ~ PhE"'
Acacia
(ph. Eur. monograph 0307)
Action and use
Bulk-fanning laxative; excipient.
When Powdered Acacia is prescribed or demanded, material
B. 6-(cyclopropylamino)-9-[(IR,4S)-4-[[(2,5-diamino-6- complying with the requirements below with the exception of
cWoropyrimidio-4-yl)oxy]methyl]cyclopent-2-enyl]-9H- Identification test A shall be dispensed or supplied.
purine-z-amine, PhE" _
DEFINITION
Air-hardened, gummy exudate flowing naturally from or
obtained by incision of the trunk and branches of Aroda
senegal L. Willd. (syn. Senegalia senegal (L.) Britton), other
species of Acacia of African origin and Acacia seyal Defile.
CHARACTERS
It is almost completely but veryslowlysoluble, afterabout
C. [(IS,4R)-4-(2,6-diamino-9H-purin-9-yl)cyclopent-2-enyJ] 2 h, in twice its mass of water leaving only a very small
methanol, residue of vegetable particles; the liquid obtained is colourless
or yellowish, dense, viscous, adhesive, translucent and weakly
acid to blue litmus paper. It is practically insoluble in ethanol
(96 per cent).
IDENTIFICATION
A. It occurs as yellowish-white, yellow or pale amber,
sometimes with a pinkish tint, friable, opaque, spheroidal,
oval or reniform pieces (tears) of a diameter from about
1-3 cm, frequently with a cracked surface, easily broken into
irregular, whitish or slightly yellowish angular fragments with
D. [(IR,4R)-4-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl] a conchoidal fracture and a glassy and transparent
cycIopent-2-enyl]methanol, appearance. In the centre of an unbroken tearthere is
sometimes a small cavity.
B. Microscopicexamination (2.8.23). The powderis white or
yellowish-white. Examine undera microscope using ethanol
(96 per cen!! R. The powder shows the following diagnostic
characters: angular, irregular, colourless, transparent
fragments. Only traces of starch or plant tissues are visible.
No stratified membrane is apparent.
C. Examine the chromatograms obtained in the test for
glucose and fructose.
E. [(IR,3S)-3-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl] Results See below the sequence of zones present in the
cycIopentyl]methanol, chromatograms obtained with reference solution (a) and the
test solution.
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1-44 Acacia 2022
Top of the plate Reference solution (a) Dissolve 5 mg of arabinose R, 5 mg of

galactose R, 5 rng of glucose R, 5 mg of rhamnose Rand 5 mg
of xylose R in 1 mL of waler R and dilute to 10.0 mL with
methanol R.
3 blue zones, very faint
Reference solulron (b) Dilute 2.5 mL of reference solution (a)
to 10.0 mL with methanol R.
Reference solutw" (e) Dissolve 5 mg of galactose Rand 5 mg
Rhamnose: a greenish-brown zone A greenish-brown zone. very faint to of gluwse R in 1 mL of water R and dilute to 10 mL with
equivalent (rhamnose) methanol R.
Intensity marker Galactose.
Xylose; a brownish-grey zone Plate TLC silica gelF'54plare R (2-10 pm).
MoMe phase waler R, acetonitrile R (15:85 VII').
Applican"on 4 J.lL of the test solution and reference
solutions (a) and (b), and 2 J.1L of reference solution (c), as
bands of 8 mm.
-- --
Development A 70 mm from the lower edge of the plate, in
an unsaturated tank.
Arabinose: a brownish-grey zone A brownish-grey zone. intense Drying A In air.
(arabinose)
Development B 70 mm from the loweredge of the plate, in
an unsaturated tank, usingfreshly prepared mobile phase.
Glucose: a greyish-blue zone Drying B In air.
Detection Treatwith a solution prepared as follows: dissolve
4 g of diphenylamine Rand 4 mL of aniline R in 160 mL of
Galactose: a greyish-blue zone A greyish-blue zone, intense
(galactose)
acetone R and add phosphoric add R until the precipitate
formed dissolvesagain (about 30 mL). Heat at 120 'C for
5-10 min and examine in daylight.
-- -~ System suitability Reference solution (c):
I or 2 brownish-grey zones. very faint - thechromatogram shows in the middle third 2 distinct
10 equivalent zones, which may be touching; the lowerzone (galactose)
and the upper zone (glucose) aregreyish-blue.
Results The chromatogram obtained. with the test solution
1 or 2 blue zones, filiIu to equivalent
showsno greyish-blue zone and no reddish zone between the
zones due to galactose and arabinose in the chromatogram
obtained with reference solution (a).
Reference solution (a) Test solution
Starch, dextr1n and agar
To 10 mL of solution S, previously boiled and cooled, add
D. Dissolve 1 g of the powdered herbal drug (355) (2.9.12) 0.1 mL of O. 05 M iodine. No blue or reddish-brown colour
in 2 mL of waterR by stirring frequently for 2 h. Add 2 mL develops.
of ethanol (96 per cen,) R. After shaking, a white gelatinous
Sterculia gum
mucilage is formed that becomes fluid upon addition of
A. Place 0.2 g of the powdered herbal drug (355) (2.9.12) in
10 mL of warer R.
a 10 mL gmund-glass-stoppered cylinder graduated in
TESTS 0.1 mL Add 10 mL of ethanol (60 per ce,,' VII') Rand
Soludon S shake. Any gel formed occupies a maximum of 1.5 mL.
Dissolve 3.0 g of the powdered herbal drug (355) (2.9.12) in B. To 1.0 g of the powdered herbal drug (355) (2.9.12) add
25 mL of water R by stirring for 30 min. Allow to stand for 100 mL of water R and shake. Add 0.1 mL of methyl red
30 min and dilute to 30 mL with water R. solution R. Not more than 5.0 mL of 0.01 M sodium hydroxide
Insoluble matter is required to change the colour of the indicator.
Maximum 0.5 per cent. Tannins
To 5.0 g of the powdered herbal drug (355) (2.9.12) add To 10 mL of solution S add 0.1 mL ei fenic chloride
100 mL of water Rand 14 mL of dilute hydrochloric add R, solution RI. A gelatinous precipitate is formed, but neither the
boil gently for 15 min, shaking frequently and filter while hot precipitate nor the liquid is dark blue.
through a tared sintered-glass lilter (2.1.2). Wash with hot Tragacanth
water R and dry at 100-105 'C. The residue weighs a Examine the chromatograms obtained in the test for glucose
maximum of 25.mg. and fructose.
Glucose and fructose Results The chromatogram obtained with the test solution
High-performance thin-layer chromatography (2.8.25). shows no faint to intense brownish-grey zone corresponding
Testsolu'ron To 0.1 g of the powdered herbal drug (355) to the zone due to xylose in the chromatogram obtained with
(2.9.12) in a thick-walled centrifuge tube, add 2 mL of a reference solution (a).
100 gIL solution of trifiuoroacetk acid R and shake vigorously. Loss on drying (2.2.32)
Stopper the tube and heat the mixture at 120 °C for 1 h. Maximum 15.0 per cent, determined on 1.000 g of the
Centrifuge, transfer 1 mL of the clear supernatant into a powdered herbal drug (355) (2.9.12) by drying in an oven at
10 mL flask and add 5 mL of methanol R. 105 'c.
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2022 Acacia 1-45
Total ash (2.4.16) Top of the plate

Maximum 4.0 per cent.
Microbial contamination
TAMC: acceptance criterion 10' CFU/g (2.6.12).
TYMC: acceptance criterion 10' CFU/g (2.6.12). 3 blue zones, very faint
Absence of Escherichia coli (2.6.11).
Absence of Salmonella (2.6.11).
Rhamnose: a greenish-brown zone A greenish-brown zone, very falnr to
FUNCTIONAliTY-RELATED CHARACTERISTICS equivalent (rhamnose)
This section provides information on characteristics that are
recognised as being relevant control parameters for one or more
Xylose: a bro....nish-grey zone
functions of the substance when usedas an excipient (see chapter
5.15). Some of the characteristics described in the Functionality-
related characteristics section may also bepresent in the mandatory
part of the monograph since they also represent mandatory quality
criteria. In such cases, a cross-reference to the tests described in the -- --
mandatorypart is included in the Functionality-related
characteristics section. Control of the characteristics can contribtue
to the qualityof a medicinal product by improving the consistency Arabinose: a brownish-grey zone A bro wnish-grey zone, intense
of the manufacturing process and the performance of the medicinal (arabinose)
product durmg use. W'hm control methods are cited, they are
recognised as being suitable for thepurpose, but othermethods can Glucose: a greyish-blue zone
also be used. Wherever results for a particular characteristic are
reponed, the control method must be indicated.
The following characteristic may be relevant for acacia usedas a Galactose: a greyish-blue zone A greyish-blue zone. intense
viscosity-increasing agentand/or suspending agentin aqueous (galactose)
preparations.
Apparent viscosity
Determine the dynamic viscosity using a capillary viscometer -- --
(2.2.9) or a rotating viscometer (2.2.10) on a 100 gIL I or 2 brownish-grey zones, very faint
to equivalent
solution of acacia (dried substance).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEII
I or 2 blue zones, faint to equivalent
Acacia, Dried Dispersion Reference solution (a) Test solution
Spray-dried Acacia C. Dissolve 1 g of the preparation to be examined in 2 mL

(Ph. Eur. monograph 0308) of waterR by stirring frequently for 20 min. Add 2 mL of
PhEIl _ ethanol (96 per e"'li R. After shaking, a white gelatinous
mucilage is formed that becomes fluid upon addition of
DEFINITION
10 mL ot oxuer R.
Powder obtained from a dispersion of Acacia (0307) after a
drying process. TESTS
Solution S
CHARACTERS
Dissolve 3.0 g of the preparation to be examined in 25 mL
It dissolves completely, after about 20 min, in twice its mass
of water R by stirring for 10 min. Allow to stand for 20 min
of water. The liquid obtained is colourless or yellowish, and dilute to 30 mL with water R.
dense, viscous, adhesive, translucent and weakly acid to blue
litmus paper. It is practically insoluble in ethanol Glucose and fructose
(96 per cent). High-performance thin-layer chromatography (2.8.25)
Test solution To 0.1 g in a thick-walled centrifuge tube add
IDENTIFICAnON
2 mL of a 100 gIL solution of rrifluoroaceti< acid R and shake
A. Examine under a microscope using ethanol (96 percent) R
vigorously. Stopper the tube and heat the mixture at 120 "C
as the mounting medium. The preparation to be examined
for I h. Centrifuge, transfer I mL of the clear supernatant
consists of predominam.ly spheroidal or irregular and angular
into a 10 mL flask and add 5 mL of methanol R.
particles varying in size (4-500 IJm), with 1 or more rounded
cavities containing 1 or several air bubbles; a few flat Reference solution (a) Dissolve 5 mg of arabinose R, 5 mg of
fragments are also present. Only traces of starch granules are galactose R) 5 mg of glucose R, 5-mg of rhamnose Rand 5 mg
visible and no plant tissue is observed. of xylose R in I mL of water R and dilute to 10.0 mL with
methanol R.
B. Examine the chromatograms obtained in the test for
glucose and fructose. Reference solution (b) Dilute 2.5 rnL of reference solution (a)
Results See below the sequence of zones present in the
chromatograms obtained with reference solution (a) and the Reference solution (c) Dissolve 5 mg of galactose Rand 5 mg
test solution. of glm:ose R in I mL of waterR and dilute to 10 mL with
methanol R.
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1-46 Acamprosate Calcium 2022
Intensity marker Galactose. FUNCTIONAUTY-RELATED CHARACTERISTICS

Place TLC ,ilica gelFm plateR (2-10 urn), This section provides itifonnatiotl on characteristics that are
Mobile phase waterR, acetomtnle R (15:85 VIV). recognised as being relevant control parameters for one or more
functions of the substance when used as an e:«ipienl (see chapter
ApplicQtion 4 j.1L of the test solution and reference 5.15). Some of the charaaeristia described in the Functionality-
solutions (a) and (b), and 2 j.1L of reference solution (c), as
related characteristics section may also be present in the mandatory
bands of 8 nun.
pan of the monograph since they also represent mandatory quality
Development A 70 mm from the lower edge of the plate, in ctiteria. In such cases, a cross-reference to lhe tests described in the
an unsaturated tank. mandatory part is imluded in the Functionality-mated
DryingA In air. characteristics section. Control of the characteristics can contribute
Development B 70 mm from the loweredge of the plate, in '0 the qualityof a medicinal produce by improving the consistency
an unsaturated tank, using freshly prepared mobilephase. of the manufacturing proms and the performance of the medicinal
DryingB In air. produce during use. W'hm control methods are cited, they are
recognised as being suitable for the purpose, but other methods can
Detection Treat with a solution prepared as follows: dissolve alsobe used. Wherever results for a particular characteristic are
4 g of dipherrylamine Rand 4 mL of anilme R in 160 mL of reported, the control method must be indicated.
acetone R and add pJwsplwric acidR until the precipitate
formed dissolves again (about 30 mL). Heat at 120°C for The following characteristic may be relevant for acacia dried
5-10 min and examine in daylight. dispersion used as a viscosiry-increasing agemandlor suspending
agent in aqueous preparations.
System suitability Reference solution (c):
- the chromatogram shows in themiddle third 2 distinct Apparent viscosity
zones, which may be touching; the lower zone (galactose) Determine the dynamic viscosity using a capillary viscometer
and the upperzone (glucose) are greyish-blue. (2.2.9) or a rotating viscometer (2.2.10) on a 100 WL
solution of acacia, dried dispersion (dried substance).
Results The chromatogram obtained with the test solution
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _~_ _ PhEII
showsno greyish-blue zone and no reddish zone between the
zones due to galactose and arabinose in the chromatogram
obtained withreference solution (a).
Starch, dextrin and agar
To 10 mL of solution S, previously boiled and cooled, add Acamprosate Calcium
0.1 mL of 0.05 M iadine. No blue or reddish-brown colour
develops.
Sterculla gum
A. Place 0.2 g in a 10 mL ground-glass-stoppered cylinder
graduated in 0.1 mL. Add 10 mL of ethanol
Ca"[H'Cr~~S031
(60 percent VIVj R and sbake. Any gel formed occupies not
more than 1.5 mL.
B. To 1.0 g add 100 mL of waterR and shake. Add 0.1 mL 400.5 77337-73-6
of methylredsolmian R. Not more than 5.0 mL of
0.01 M sodium hydroxide is required to change the colour of Action and use
the indicator. Treatment of alcoholism.
TannIns Preparation
To 10 mL of solution S add 0.1 mL offerric chloride Acamprosete Gastro-reslatant Tablets
solution RI. A gelatinous precipitate is formed, but neither the PhEII _
precipitate nor the liquid is dark blue.
Tragacanth DEFINITlON
Examine the chromatograms obtained in the test forglucose Calciwn bis(3-acetamidopropane-l-sulfonate).
and fructose. Content
Results The chromatogram obtained with the test solution 98.0 per cent. to 102.0 per cell'. (dried substance).
shows no faint to intense brownish-grey zone corresponding CHARACTERS
to the zone due to xylose in the chromatogram obtained with Appearance
reference solution (a). White or almost white powder.
Loss on drying (2.2.32) Solubillty
Maximum 10.0 per cent, detennined on 1.000 g by drying in Freely soluble in water, practically insoluble in ethanol
an oven at 105°C. (96 per cent) and in methylene chloride.
Total ash (2.4.16) IDENTIFICATION
Maximum 4.0 per cent A. Infrared absorption spectrophotometry (2.2.24).
Microbial contamination Comparison acamprosate calcium CRS.
TAMC: acceptance criterion 10' CFU/g (2.6.12).
B. It gives reaction (a) of calcium (Z.3.1).
TYMC: acceptance criterion 10' CFU/g (2.6.12).
TESTS
Absence of Escherichia cdi (2.6.13).
Solution S
Absence of Salmonella (2.6.13). Dissolve 5.0 g in carbon dioxide-free water R and dilute to
100 mL with the same solvent.
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2022 Acarnprosare Calcium 1-47
Appearance of solution - stationaryphase: end-copped oaadecylsilyl silica gelfor

Solution S is clear (2.2.1) and colourless (2.2.2, Method If). chromatography compatible with 100 per cent aqueaus mobile
pH (2.2.3) phases R (5 um),
55 to 7.0 for solution S. Mobile phase Mix 5 mL of triethylamine R and about
Impurity A 900 mL of waUir for chromatagraphy R, adjust to pH 4.0 with
Liquid chromatography (2.2.29). phosphoric acid R and dilute to 1000 mL with waUlI' for
chromatography R.
Tesc solution Dissolve 0.400 g of the substance to be
examined in distiUed water R and dilute to 20.0 mL with the Flow raUl 0.7 mUmin.
same solvent. Dilute 10.0 mL of the solution to 100.0 mL Detection Spectrophotometer at 210 om.
with borate blfffer solurion pH 10.4 R. Introduce 3.0 mL of this Injection 20 ilL of test solution (a) and reference
solution into a 25 mL ground-glass-stoppered tube and add solutions (a) and (e).
0.15 mL of a freshly prepared 5 gIL solution of Run time 2.5 times the retention time of acamprosate.
ftuorescamine R in acetoniuile R. Shake immediately and Identification 0/ impun·,ies Use the chromatogram obtained
vigorously for 30 s. Heat in a water-bath at 50°C for
with reference solution (c) to identify the peakdue to
30 min. Cool undera stream of cold water. Centrifuge and impurity B.
filter the supernatant through a membrane filter (nominal
pore size 0.45 urn). Rdative retention With reference to acamprosare (retention
time e about 9 min): calcium ;;;; about OAj
Reference solution Dissolve 50.0 mg of acamprosau impurity B = about 0.8.
impurity A CRS in disti/led waUir R and dilute to 200.0 mL
with the same solvent. Dilute 0.4 mL of me solution to System suitability Reference solution (c):
~ resolution: minimum 5.0 between the peaks due to
100.0 mL with borace buffersolution pH lOA R. Iotroduce
3.0 mL of this solution into a 25 mL ground-glass-stoppered impurity Band acamprosate.
tube. Proceed as described for the test solution, starting from Calculation ofpercentage contents:
'and add 0.15 mL of a freshly prepared 5 gIL solution of - for each impurity, use the concentration of acamprosate
fiuarescamine R J
•
calcium in reference solution (a).
Column: Limits:
- size: 1 == 0.15 m,0 = 4.6 mm; - unspecified impurities: for each impurity, maximum
- stationary phase: end-capped oaadecyisi/yl silira gelfor 0.05 per cent;
chromatagraphy R (5 urn). - total: maximum 0.3 per cent;
Mobilephase a"tanilril. R, methanolR, 0.1 M phosphate - reporting threshold: 0.03 per cent; disregard the peak due to
buffersolution pH 6.5 R (10:10:80 VIVIV). calcium.
Flow raUl 1 mIJrnin. Loss on drying (2.2.32)
Maximum 0.4 per cent, determined on 1.000 g by drying in
Detection Spectrophotometer at 261 DOl.
an oven at 105°C.
Inje<rion 20 pL.
ASSAY
Run time 6 times the retention time of impurity A
Liquid chromatography (2.2.29) as described in the test for
derivative.
related substances with the following modification.
Retention time Fluorescamlne e about 4 min; impurity A
derivatfve » about 8 min; acamprosate is not detectedby this
Iniecuon 20 pI. of test solution (b) and reference
solution (b).
system,
Limit: Calculate the content of CtoH20CaN20SS2 taking into
account the assigned content of acamprosau calcium CRS.
- impun·ty A: not more than the area of the corresponding
peak in the chromatogram obtained with the reference IMPURITffiS
solution (0.05 per cent). Specified impurities A.
Related substances Otherdetectable impurities (the following substances would, if
Liquid chromatography (2.2.29). present a1 a sufficient level, be detected by oneor other of the tests
Testsolution (a) Dissolve 0.100 g of the substance to be in the monograph. They am limited by the general acceptance
examined in 8 mL of water R using sonication and dilute to cruenon for other/unspecified impurities and/or by thegeneral
10.0 mL with the same solvent. monograph Substances for pharmaceutical use (2034). It is
therefore not necessary to identify these impurities for
Test solution (b) Dilute 3.0 mL of test solution (a) to
demonstration of compliance. See also 5.10. Control 0/ impurities
100.0 mL with waUlI' R.
in subsronces for pharmaceutical use) B, c.
Reference solution (a) Dilute 1.0 mL of test solution (a) to
100.0 mL with waUir R. Dilute 1.0 mL of this solution to
20.0 mL with waUir R.
Reference solution (b) Dissolve 30.0 rng of acamprosate
A. 3-aminopropane-I-sulfonic acid (homoraurine),
calcium CRS in 20 mL of water R using sonication and dilute
to 100.0 mL with the same solvent.
Reference solution (c) Dissolve 10 mg of calcium bis(fonny/
homotaurine) R (corresponding to about 9 mg of impurity B)
in 1 mL of test solution (a) and dilute to 100 mL with B. 3-fonnamidopropane-l-sulfonic acid (formyl
wmer R. homoraurine),
Column:
- size: I;;;; 0.25 m, 0 = 4.6 mm;
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1-48 Acarbose 2022
Test solution Dissolve 0.200 g of the substance to be

examined in water R and dilute to 10.0 mL with the same
solvent.
Reference solution (a) Dissolvethe contents of a vial of
C. 3-(N-methylacetamido)propane-l-sulConic acid. acarbose GRS in 5.0 mL of water R.
__ ~ PhE<6 Reference solution (b) Dissolve me contents of a vial of
aCQrbose for peak identification GRS (acarbose containing
impurities A, B, CJ DJ EJ F and G) in 1 mL of water R.
Reference solution (c) Dilute 1.0 mL of the test solution to
Acarbose 100.0 mL with waterR.
Column:
(ph. Eur. monograph 2089) - size: 1= 0.25 m, 0 = 4 mmj
- stationary phase: aminopropylsily1 sil",a gelfor
chromatography R (5 1IJI1);
- temperoture: 35 "C.
klobilephase Mix 750 volumes of acetonitrile Rl and
250 volumes of a solution containing 0.60 gIL of potassium
dihydrogen phosphate Rand 0.35 gIL of disodium hydrogen
phosphate dihydrate R.
Flow .rote 2.0 mllmin.
c"H,,NO,, 646 56181)-94-0
Detection Spectrophotometer at 210 run.
Action and use lnjution 10 ~L of the test solution andreference
Alpha-glucosidase inhibitor; treatment of diabetes mellitus. solutions (b) and (c).
PhE" _ Run time 2.5 times the retention timeof acarbose.
Idemification of impun'ties Use the chromatogram supplied
DEFINITION with acarbose for peak idenufication CRS and the
0-4,6-Dideoxy-4-[[(IS,4R,5S,6S)-4,5,6-trihydroxy-3- chromatogram obtained with reference solution (b) to
(hydroxymethyl)cyclohex-2-enyl]amino]-«-D-glucopyranosyl- identify the peaks due to impurities AJ BJ C, D, E, F and G.
(I ~4)-0-«-l}-glucopyranosyl-(1 ~ 4)-D-glucopyranose, which
Relative retention With reference to acarbose (retention
is produced by certain strains of Aetinoplanes utonensis.
Content = =
impurity B about 0.8; impurity A about 0.9;
95.0 per cent to 102.0 per cent (anhydrous substance). impurity C = about 1.2j impurity E = about 1.7;
CHARACTERS impurity F = about 1.9; impurity G = about 2.2.
Appearance System suitability Reference solution (b):
While or yellowish, hygroscopic, amorphous powder. - the chromatogram obtained is similar to the
Solubility chromatogram supplied with euarbose for peak
Very soluble in water, soluble in methanol, practically identification CRS;
insoluble in methylene chloride. - peak-to-'llalley ratio: minimum 1.2, where H p = height
above the baseline of the peak due to impurity A and
IDENTIFICATION HI} = heightabove the baseline of the lowestpointof the
A. Infrared absorption spectrophotometry (2.2.24). curve separating this peak from the peak due to acarbose.
Comparison arorbose for identification CRS. Limits:
B. Examine the chromatograms obtained in the assay. - correction faaon: for the calculation of content, multiply
Results The principal peak in the chromatogram obtained the peak areas of the Collowing impurities by the
with the test solution is similar in retention time and size to corresponding correction factor: impurity B = 0.63j
the principal peak in the chromatogram obtained with = =
impurity D 0.75; impurity E 1.25; impurity F 1.25; =
reference solution (a). impurity G" 1.25;
- impuniy G: not more than 1.5 times the area of the
TESTS principal peak in the chromatogram obtained with
Solution S reference solution (c) (1.5 per cent);
Dissolve I. 00 g in carbon dioxide-free water R and dilute to - impurity D: not more than the area of the principal peak
20.0 mL with the same solvent. in the chromatogram obtained with reference solution (c)
pH (2.2.3) (1.0 per cent);
5.5 to 7.5 for solution S. - impurity A: not more than 0.6 times the area of the
Specific optical rotation (2.2.7) principal peak in the chromatogram obtained with
+ 168 to + 183 (anhydrous substance). reference solution (c) (0.6 per cent);
- impwilY B: not more than 0.5 times the area of the
Dilute 2.0 mL oC solution S to 10.0 mL with water R.
principal peakin the chromatogram obtained with
Absorbance (2.2.25) reference solution (c) (0.5 per cent);
Maximum 0.15 at 425 nm Cor solution S. - impurities F, G: for each impurity, not more than
Related substances 0.3 times the area of the principal peak in the
liquid chromatography (2.2.29). chromatogram obtained with reference solution (c)
(0.3 per cent);
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j
2022 Acarbose 1-49
- impurity E: not more than 0.2 times the area of the HO

principal peak in the chromatogram obtained with o
reference solution (c) (0.2 per cent);
- a,ry other impurity. for each impurity) not more than HO
0.2 times the area of the principal peak in the
chromatogram obtained with reference solution (c) HO~ CH, H~O
OH
-
(0.2 per cent);
not more than 3 times the area of the principal peak
lOlal: 0-;- ~o\.. OH 0
in the chromatogram obtained withreference solution (c)
(3.0 per cent);
HO ~~o 0
OH OH OH
- disregard limit: 0.1 times the area of the principal peak in
the chromatogram obtained with reference solution (c) C. Il-D-g1ucopyranosyI4-G-[4,6-dideoxy-4-[[(IS,4R,5S,6S)-
(0.1 per cent). 4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-euyll
Water (2.5.12) amino] -c-o-glucopyranosyl]-e-n-glucopyrancside,
Maximum 4.0 per cent, determined on 0.300 g.
Sulfated ash (2.4.14) HO~ CH, H~O
10;- ~HO
Maximum 0.2 per cent} determined on 1.0 g. O
OH OH
ASSAY
HO N 0
Liquid chromatography (2.2.29) as described in the test for H
OH OH OH
Injection Test solution and reference solution (a). D. 4-G-[4,6-dideoxy-4-[[(I S,4R,5S,6S) -4,5,6-trihydroxy-3-
Calculate the percentage content OfC2)H4~018 raking inro (hydroxymethyl)cyclohex-2-enyI}amino)-<1.-D-
account the assigned content of acarbose CRS. glucopyranosylj-n-glucopyranose,
STORAGE
r
OH
In an airtight container, HO HO HO HO
IMPURITIES
Specified impuriues A, B, C, D, E, F, G.
-~ )2-0\ '-~o,-~f OH
Otherdetectable impurities (the following substances would, if
present at a sufficient level, bedetected by oneor other of the tests
H~~~/4'iYL(\0
OH OH OH OH
in the monograph. They are limited by thegeneral acceptance
criterion for other/unspecified impurities. It is therefore not E. G-4,6-dideoxy-4-[[(IS,4R,5S,6S)-4,5,6-trihydroxy-3-
necessary UJ identify these impurities for demonstration of (hydroxymethyl)cyclohex-2-enyl]amino}-<1.-D-
compliance. See also 5.10. Control of impurities in subsumces for glucopyranosyl-H-» 4)- O-<t.-D-g1 ucopyranosyl-( 1-->4)- O-<t.-
pharmaceutical use) H. n-glucopyranosyl- (I ~ 4)-D-arabin<>-hex- 2-ulopyranose
(4-Q...a-acarbosyl-D-frucropyranose),
F. G-4,6-dideoxy-4-[[(IS,4R,5S,6S)-4,5,6-trihydroxy-3-
A. 0-4,6-dideoxy-4-[[(IS,4R,5S,6S)-4,5,6-trihydroxy-3- (hydroxymethyl)cyclohex-2-enyl]amino)-<1.-D-
(hydroxymethyl)cyclohex-2-enyl}amino]-<1.-o- g1ucopyranosyl-(l ~4)-O-<t.-D-g1ucopyranosyl-(I-->4)-O-<t.
g1ucopyranosyl-(l ~ 4)-O-<t.-o-g1ucopyranosyl-(1--> 4)-D- n-glucopyranosyl-It ~4)-o-g1ucopyranose (4-0-<1.-
arabino-hex-2-ulopyranose, acarbosyl-n-glucopyranose),
HO
HO~ CH, 0 HhO H~O
_OH
~o'\
_ 0
OH OH OH
HO ~~o 0 HO
~C>HhO H~O
OH OH OH OH
HO OH OH OOH 0
B. (IR,4R,5S,6R)-4,5,6-trihydroxy-2-(hydroxymethyl)
cyclohex-2-enyI4-0-[4,6-dideoxy-4-[[(IS,4R,5S,6S)- HOb H N 0 0 0
4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl) H
OH OH OH OH
amino] -«-o-glucopyranosyl] -c-n-glucopyrenoside,
G. c-n-gluccpyranosyl G-4,6-dideoxy-4-[[(lS,4R,5S,6S)-
4,5,6-trihydroxy-3-(hydroxymethyl)cyclohex-2-enyl)
amino]-<1.-n-gtucopyranosyi-tt-« 4)-O-<t.-D-g1ucopyranosyl-
(I ~4)-O-<t.-D-g1ucopyranoside(c-n-glucopyrancsyl
c-acarboside),
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I-50 Acebutolol Hydrochloride 2022
Comparison acebutolol hydrochloritk CRS.

C. Thin-layer chromatography (2.2.27).
examined in methanol R anddilute to 20 mL with the same
solvent.
Reference solution (a) Dissolve 20 mg of acebutolol
H.O-4,6-dideoxy-4-[[(IS,4R,5S,6S)-4,5,6-ttihydroxy-3- hydrochloride CRS in methanol R and dilute to 20 mL with the
(hydroxymethyl)cyclohex-2-enyl)amino)-<x-o- same solvent. .
glucopyranosyl-(I ;4)-D-6-deoxy-<x-o-glucopyranosyl- Reference solution (b) Dissolve 20 mg of pindolol CRS in
(1-->4)-o-glucopyranose. methanol R and dilute to 20 mL with the samesolvent.
____ ~~ ~~ ""61 To 1 mL of this solution add 1 mL of reference solution (3).
Plate TLC silica gelFm plat< R.
Mobile phase perchloric acidR, methanol R, water R
(5:395:600 VIVIV).
Acebutolol Hydrochloride ***
*** *** Application 10 pL.
(Ph. Eur. monograph 0871) *** Development Over 314 of the plate.
Drying In air.
o CH,
Detection Examine in ultraviolet light at 254 om.
E
H pH H
o ~~ O~NyCH, ·Ha System suitability The chromatogram obtained with

reference solution (b) shows2 clearly separated principal
HC~N.# CH3 spots.
3 H and enanuceer
Results The principal spot in the chromatogram obtained
with the test solution is similar in position and size to the
372.9 34381-68-5 principal spot in the chromatogram obtained with reference
solution (a).
Action and use
Beta-adrenoceptor antagonist. D. It gives reaction (a) of chlorides (2.3.1).
Preparations TESTS
Acebutolol Capsules Appearance of solution
Acebutolol Tablets The solution is not more opalescent than reference
suspension II (2.2.1) and not more intensely coloured than
""'v _ reference solution BY, (2.2.2, Method If).
DEFINITION Dissolve 0.5 g in wa"r R and dilute to IO mL with the same
N-[3-Acetyl-4-[(2RS)-2-hydroxy-3-[(I-methylethyl)amino] solvent.
propoxy)phenyl]butanamide hydrochloride. pH (2.2.3)
Content 5.0 to 7.0.
99.0 per cent to 101.0 per cent (dried substance). Dissolve 0.20 g in carbon dioxitk-free water R and dilute to
CHARACTERS 20 mL with the same solvent.
Appearance Related substances
Whiteor almost white, crystalline powder. Liquid chromatography (2.2.29).
Solubility Test solution Dissolve 0.100 g of the substance to be
Freely soluble in water and in ethanol (96 per cent), very examined in mobile phase A and dilute to 50.0 mL with
slightly soluble in acetone and in methylene chloride. mobile phase A.
mp Reference solution (a) Dissolve 20.0 mg of the substance to
About 143 'C. be examined in mobile phase A and dilute to 100.0 mL with
mobile phase A. Dilute 0.5 mL of this solution to 50.0 mL
IDENTIFICATION with mobile phase A.
First idemificaeion: B, D. Reference solution (b) Dissolve the contents of a vial of
Second identification: A, C, D. acebutolol impurity I CRS in 1.0 mL of mobile phase A.
A. Ultraviolet and visible absorption spectrophotometry Reference solulion (c) Mix 2.0 mL of reference solution (a)
(2.2.25). and 1.0 mL of reference solution (b) and dilute to 10.0 mL
Test solution Dissolve 20.0 mg in a 0.1 per cent VIV with mobile phase A.
solution of hydrochloric acid R and dilute to 100.0 mL with Reference solurion (d) Dissolve 5.0 mg of acebutolol
the same acid solution. Dilute 5.0 mL of this solution to impurity C CRS in 10 mL of acetonitrile R and dilute to
100.0 mL with a 0.1 per cent VlVsolution of hydrochloric 25.0 mL with mobile phase A. Dilute 0.5 mL of this solution
acid R. to 50.0 mL with mobile phase A.
Spe<tral range 220-350 urn. Reference rolution (e) Dissolve 5.0 mg of acebutolol
Absorption maxima At 233 urn and 322 nm. impurity B CRS in 10.0 mL of acetonitrik R and dilute to
Specific absorbance at the absorption maximum 555 to 605 at 25.0 mL with mobile phase A. Dilute 1.0 mL of this solution
233 urn. to 50.0 mL with mobile phase A.
B. Infrared absorption spectrophotometry (2.2.24). Column:
- size: 1:= 0.125 m, 0 = 4 DUD,
Preparation Discs.
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2022 Acebutolol Hydrochloride I-51
-
stationary phase: end-capped oetaduylsilyl silica gel for
o CH,
chromatography R (5 urn),
- temperature: 40 "C.
Mobile phase:
- mobile phase A: mix 2.0 mL of phosphoric acid R, and
3.0 mL of triethylamine R and dilute to 1000 mL with
H3C~NH
o
EI "" 006
d
anO eoenuomer
A. N-[3-acetyl-4-[(2RSJ-oxirnn-2-ylmethoxy]
water R;
phenyl]butanamide,
- mobile phase B: mix equal volumes of acetonitrile Rand
mobile phase Ai o CH,
Time
(mln)
0-2
2 - 30.5
30.5·41
Mobile phase A
(per cent VIJ?
98
98
->
10
10
MobUe phase B
(per cent J'fIJ)
2
2 ...... 90
90
W
H,C...-"-.~ E
I
~ O~N
'"
H pH H
Y
CH,
CH3
and enanliomer
B. N-[3-acetyl-4-[(2RSJ-2-hydroxy-3-[(I-methylethyl)amino]
propoxyJphenyl]acetamide (diacetolol),
Flow rate 1.2 mUmin.
Detection Spectrophotometer at 240 nm. o CH,
E
Injection 25 ~L.
System suitability Reference solution (c): "" OH
o I
- resolution: minimwn 7.0 between the peaks due to
impurity I and acebutolol.
HC~N
a H
d
Limits:
- impurity B: not more than the area of the principal peak in C. N-(3-acetyl-4-hydroxyphenyl)butanamide,
the chromatogram obtained with reference solution (e)
(0.2 per cent);
- impun"ty C: not more than the area of the principal peak
in the chromatogram obtained with reference solution (d)
(0.1 per cent);
- lmpun"ty 1: not more than twice the area of the principal
peak in the chromatogram obtained with reference
solution (a) (0.2 per cent); D.I-[5-amino-2-[(2RSJ-2-hydroxy-3-[(I-methylethyl)amino]
- any other impun·ty: for each impurity, not more than the propoxy)phenyl]ethanone,
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.1 percent); H OH
O
- total: not more than 5 times the area of the principal peak O~ ~ yCH'
in the chromatogram obtained with reference solution (a) ~ I· CH and enanUomer
(0.5 per cent); H,C~~ '" a

- disregard limil: 0.5 times the area of the principal peak in
the chromatogram obtained with reference solution (a)
E. N-[4-[(2RSJ-2-hydroxy-3-[(I-methylethyl)amino]propOXY]
(0.05 per cent).
phenyl)butanamide,
L088 on drying (2.2.32)
Maximum 0.5 per cent, determined on 1.000 g by drying in o CH, H OH
E
an oven at 105°C for 3 h.
I ~ ~
o .: OH
Sulfated ash (2.4.14) o andenanliomer
H'C~~
ASSAY
Dissolve 0.300 g in 50 mL of ethanol (96 P<' cenlJ R and add
I mL of 0.1 M hydrochloric acid. Carry out a potentiometric F. N-[3-acetyl-4-[(2RSJ-2,3-dihydroxypropoxy]
titration (2.2.20), using 0.1 M sodium hydroxide. Read the phenyljbutanamide,
volume added between the 2 points of inflexion.
I mL of 0.1 M sodium hydroxide is equivalent to 37.29 mg of
ClsH29C1N204·
STORAGE
Protected from light.
IMPURITIES
Specified impurities A, BJ CJ DJ EJ FJ OJ HJ 1J JJ K.
G. N,N' -[[(I -methylethyl)imino]bi8[(2-hydroxypropane-1 ,3-
diyl)oxy(3-acetyl-I,4-phenylene)]]dibutanamide (biarnine),
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I-52 Aceclofenac 2022
Solubility
Practically insoluble in water, freely soluble in acetone,
soluble in ethanol (96 per cent).
IDENfIFICATION
First identification: B.
Second identification: A, C.
H. N)v'-[(2-hydroxypropane-1 ,3-diyl)bis[0X)'(3-acetyl-1 ,4- A. Ultraviolet and visible absorption spectrophotometry
phenylene)]]dibutanamide, (2.2.25).
Testsolution Dissolve 50.0 mg in methanol R and dilute to
100.0 rnL with the same solvent. Dilute 2.0 mL of the
solution to 50.0 rnL with methanol R.
Spearal range 220-370 run.
Absorption maximum 275 run.
Specific absorbance at theabsorption maximum 320 to 350.
I. N- [3-acetyl-4- [(2RS)- 3-(ethylamino)- z-hydroxypropoxyj B. Infrared absorption spectrophotometry (2.2.24).
phenyl]butanamide, Comparison: Ph. Bur. reference spectrum of ocedcfenac.
C. Dissolve about 10 mg in 10 rnL of ethonol (96 percem) R.
To I mL of the solution, add 0.2 mL of a mixture, prepared
immediately beforeuse, of equal volumes of a 6 gIL solution
of potassium fenicyan ide R and a 9 gIL solution of ferric
chloride R. Allow to stand protected from light for 5 min.
Add 3 mL of a 10.0 gIL solution of hydrochlori< add R. Allow
to stand protected from light for 15 min. A blue colour
J. N-[3-acetyl-4-[(2RS)-2-hydroX)'-3-[(I-methylethyl)aminoj develops and a precipitate is formed,
propoxyjphenyljprcpanamlde, TESTS
H'W
C=&I '
Related substances
H pH H Liquid chromatography (2.2.29). Prepore the solutWns
'" O~N immediarely before use.
Y CHo and enanliomer Solvent mixture Mobile phase A, mobilephaseB
HC~N.# CH3 (30:70 VIV).
, H
Test solution Dissolve 50.0 mg of the substance to be
K. N-[3-butanoyl-4-[(2RS)-2-hydroX)'-3-[(1- examined in the solventmixture and dilute to 25.0 mL with
methylethyl)amino]propoX)'jphenyljbutanamide. the solvent mixture.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POElr Reference solurian (a) Dissolve 21.6 mg of didofenae
sodium CRS (impurity A) in the solvent mixture and dilute 10
50.0 rnL with the solvent mixture.
Reference sdution (b) Dilute 2.0 rnL of the test solution to
Aceclofenac
Reference solution (c) Mix 1.0 rnL of reference solution (a)
(ph. Bur. monograph 1281) and 1.0 rnL of reference solution (b) and dilute to 100.0 rnL
with the solvent mixture.
Reference solntum (d) Dissolve 4.0 mg of acedofenae
impun·ty F CRS in the solventmixture and dilute to 10.0 mL
Reference solution (e) Dissolve 2.0 mg of acedofenac
impurity H CRS in the solvent mixture and dilute to 10.0 rnL
Reference solution (f) Mix 1.0 rnL of reference solution (b),
354.2 89796-99-6 1.0 rnL of reference solution (d) and 1.0 rnL of reference
solution (e) and dilute to 100.0 mL with the solventmixture.
Action and use
Cycle-oxygenase inhibitor; analgesic; anti-inflammatory.
Reference solnt"," (g) Dissolve 5.0 mg of acedofenac
impurity I CRS in the solventmixture and dilute to 50.0 mL
POE" ~_---------- with solvent mixture. Dilute 1.0 mL of the solution to
50.0 mL with the solvent mixture.
DEFINITION
[[[2- [(2,6- DicWorophenyl)amino] phenyl] acetyl] oX)'j acetic Reference solutinn (h) Dissolve 4 mg of acedofenac for peok
acid. idenrijication CRS (containing impurities B, C, D, E and G)
in 2 mL of the solventmixture.
Content
Column:
99.0 per cent to 101.0 per cent (dried substance).
- size: 1= 0.25 m, 0 = 4.6 mm;
CHARACTERS
Appearance
White or almost white, crystalline powder.
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2022 Aceclofenac I-53
- stationary phase: spherical end-capped octadecy/sily/ silica gel Loss on drying (2.2.32)
for chromatography R (5 urn) with a pore size of 10 om Maximum 0.5 per cent, determined on 1.000 g by drying in
and a carbon loading of 19 per cent; an oven at 105 "C.
- temperature: 40 "C. Sulfated ash (2.4.14)
MobJe phase: Maximum 0.1 per cent, determined on 1.0 g.
- mobile phase A: 1.12 gIL solution of phosphoric acidR
ASSAY
adjusted to pH 7.0 with a 42 gIL solution of sodium
Dissolve 0.300 g in 40 mL of methanol R. Titrate with 0.1 1\1
hydroxide R;
- mobile phase B: waterR, acetonitrile R (10:90 VIV); sodium hydroxide, determining the end-point
potentiometrically (2.2.2rJ).
Time Mobile phase A Mobile phase B 1 mL of 0.1 1\-1 sodium hydroxide is equivalent to 35.42 mg of
(min) (per cent VIP) (per cent JIm C,oH 13CI,N04 ·
0-25 70 50 30 --> 50 STORAGE
25 - 30 50 20 50 --> 80
30 - 50 20 80
IMPURITIES
Specified impurities A, B, C, D, E, F, G, H, I.
Injection 10 JIL of the test solution and reference
solutions (c), (d), (e), (I), (g) and (h).
0(
""
1
NH
Co,H
Identification of impurities Use the chromatogram obtained CI~CI

with reference solution (c) to identify the peak due to
impurity Aj use the chromatogram supplied with acedofenoc
U
for peak identification CRS and the chromatogram obtained A. [2-[(2,6-dichlorophenyl)antino]phenyl]acetic acid
with reference solution (h) to identify the peaks due to (diclofenac),
impurities B, C, D, E and G; use the chromatogram
obtained with reference solution (d) to identify the peak due
to impurity F; use the chromatogram obtained with reference
solution (e) to identify the peak due to impurity H; use the
chromatogram obtained with reference solution (g) to
identify the peak due to impurity I.
Relative retention With reference to aceclofenac (retention
= =
time about 11 min): impurity A about 0.8;
= =
impurity G about 1.3j impurity H about 1.5j B. methyl [2-[(2,6-dichlorophenyl)antino]phenyl]acetate
impurity I =about 2.3; impurity D ;;;; about 3.1; (methyl ester of diclofenac),
impurity B ;;;; about 3.2j impurity E ;;;; about 3.3j
impurity C ;;;; about 3.5; impurity F;;;; about 3.7.
- resolution: minimum 5.0 between the peaks due to
impurity A and acedofenac.
ocr°
""
I
CI~CI
NH
O.............. CH3
Limits:
- impun·ly A: not more than the area of the corresponding
U
peak in the chromatogram obtained with reference C. ethyl [2-[(2,6-<1ichlorophenyl)amino]phenyl]acetate (ethyl
solution (c) (0.2 per cent); ester of diclofenac),
- impurities B, C, D, E, G: for each impurity, not more than
the area of the peak due to aceclofenac in the a
chromatogram obtained with reference solution (f)
(0.2 per cenr);
r""(Y°J OCH,
- impuniy F: not more than the area of the corresponding V"NH O

peak in the chromatogram obtained with reference CI~CI
solution (I) (0.2 per cent);
- impun·ty H: not more than 1.5 times the area of the U
corresponding peak in the chromatogram obtained with
reference solution (f) (0.15 per cent); D. methyl [[[2-[(2,6-dichlorophenyl)amino]phenyl]acetyl]oxy]
- impurity I: not more than 1.5 times the area of the acetate (methyl ester of aceclofenac),
reference solution (g) (0.15 per cent); °
r""(Y°J
- unspecified impurities: for each impurity) not more than O »<; CH,
0.5 times the area of the peak due to aceclofenac in the
chromatogram obtained with reference solution (f)
V"NH O
(0.10 per cent); CI~CI
- total: maximum 0.7 per cent,
- disregard limit: 0.25 times the area of the peak due to
U
aceclofenac in the chromatogram obtained with reference E. ethyl [[[2-[(2,6-dichlorophenyl)amino]phenyl]acetyl]oxy]
solution (I) (0.05 per cent). acetate (ethyl ester of aceclofenac),
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I-54 Acemetacin 2022
CHARACTERS
Appearance
Yellow or greenish-yellow, crystalline powder.
Solubility
Practically insoluble in water, soluble in acetone, slightly
soluble in anhydrous ethanol.
It shows polymorphism (5.9).
F. benzyl [[[2-1(2,6-<1ichlorophenyl)amino]phenyl]acetyl]oxy] IDENTIFICATION
acetate (benzyl ester of aceclofenac), Infrared absorption spectrophotometry (2.2.24).
Comparison acemetacin CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and me reference
substance separately in aaWne R, evaporate to dryness and
record new spectra using the residues.
TESTS
Related substances
Liquid chromatography (2.2.29).
G. [[1[[2-[(2,6-dichlorophenyl)amino]phenyl]acetyl]oxy]
Testsolution Dissolve 0.100 g of die substance to be
acetyl]oxy]acetic acid (acetic aceclofenac),
examined in acetonitrile for chromatography R and dilute to
20.0 mL with the same solvent.
Reference solution (a) Dilute 5.0 mL of the test solution to
50.0 mL with acetonitrile for chromatography R. Dilute 1.0 mL
of this solution to 100.0 mL with acetonitrile for
chromatography R.
Reference solution (b) Dissolve 5.0 mg of acemetacin
impurity A CRS and 10.0 mg of indometaein CRS
(impurity B) in acetonitrile for chromatography R, and dilute to
H. III[[[[2-[(2,6-dichlorophenyI) amino]phenyl]acetyl]oxy] 50.0 mL with the samesolvent.
acetyl]oxy]acetyl]oxy]acetic acid (diacetic aceclofenac), Reference solution (c) Dilute 1.0 mL of reference solution (b)
to 20.0 mL with automin'le for chromatography R.
Reference solution (d) To 1 mL of reference solution (b),
add 10 mL of the test solution and dilute to 20 mL with
acetonitrile for chromatography R.
Reference solution (e) Dissolve the contents of a vial of
acemetacin impun'ly mixture CRS (containing impurities C, D,
E and F) in 1.0 mL of the lest solution.
I. 1-(2,6-dichlorophenyl)-1,3-dihydro-2H-indol-2-one.
Column:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pf>E"
- size: I::::: 0.25 m, 0 = 4 nun;
- stationary phase: spherical end-capped octadecy1si/yl silica gel
for chromatography R (5 pm);
Acemetacin Mob.e phase:
- mobile phaseA: dissolve 1.0 g of potassium dihydrogen
(Ph. Eur. monograph 1686) phosphate R in 900 mL of waterR, adjust to pH 6.5 with
I M sodium hydroxide and dilute to 1000 mL with
waterR;
- mob.e phase B: acetonitrile for chromatography R;
Time Mobile phase A Mobile phase B

(min) (per cent VM (per cent V/1?
0-5 .5 5
5-' 95 --) 65 5 -. 35
415.8 53164-05·9 9·16 65 35
16 - 28 65 -. 20 35 -. 80
Action and use 28 - 34 20 80
Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.
Pf>E" _ Flow rat< 1.0 mlJmin.
Detection Spectrophotometer at 235 nm.
DEFINITION
Injection 20 1'1.
[[[I-(4-Chlorobenzoyl)-5-methoxy-2-methyl-IH-indol-3-yl]
acetyl]oxy]acetic acid. Identification of impurities:
- use the chromatogram supplied with acemetacin
Content
impun'ty mixture CRS and the chromatogram obtained
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2022 Acemetacin I-55
with reference solution (e) to identify the peaks due to

impurities C, D, E and F;
- use the chromatogram obtained with reference
solution (b) to identify the peak due to impurity B.
Relative retention With reference to acemetacin (retention
time e about 15 min): impurity A = about 0.7;
impurity B ;;;; about 0.9; impurity F = about 1.2,;
impurity C = abcur 1.3; impurity D = about 1.5; B. [1-(4-ehlorobenzoyl)-5-methoxy-2-methytindol-3-yl]acetic
=
impurity E about 2.2. acid (indometacin),
System suitabl7ilY Reference solution (d):
- peah-to-oalley ratio: minimum 15, where Hp = height
above the baseline of the peak due to impurity Band
H" = height above the baselineof the lowest point of the
curve separating this peakfrom the peak due [Q
acernetacin.
Limits:
- correaion factors: for the calculation of content, multiply
the peak areas of the following impurities by the C. [[[1-(3,4-dichlorobenzoyl)-5-methoxy-2-methyl-1 H-indol-
corresponding correction factor: impurity C = J.3; 3-yl]acetyl]oxy]acetic acid,
impurity D =
1.4; impurity F 1.3;=
- impurity E: not more than 3 times the area of the principal
solution (a) (0.3 per cent);
- impuniy B: not more than the area of the corresponding
solution (c) (0.2 per cent);
- impun·ty A: not more than the area of the corresponding
- impurities C} D} F: for each impurity} not more than the D. [[[ 1-(4-chlorobenzoyl)-6-(I, I-dimethyl ethyl)-5-methoxy-2-
area of the principal peak in the chromatogram obtained methyl-IH-lndol-3-yl]acetyl]oxy]acetic acid,
with reference solution (a) (0.1 per cent);
- unspecified impurities: for each impurity} not more than the
- total: not more than 4 times the area of the principal peak
in the chromatogram obtained with reference solution (a)
(0.4 per cent);
the chromatogram obtainedwith reference solution (a)
(0.05 per cent). E. I, l-dimethylethyl [[[1-(4-chlorobenzoyl)-5-methoxy-2-
methyl-IH-indol-3-yl]acetyl]oxy[acetate,
Loss on drying (2.2.32)
an oven at 105 "C.
Sulfated ash (2.4.14)
ASSAY
Dissolve 0.350 g in 20 mL of acetone R aod add 10 mL of
water R. Titrate with 0.1 AI sodium hydroxide, determining the
end-point potentiometrically (2.2.20).
F. [[[[[I-(4-chlorobenzoyl)-5-methoxy-2-methyl-IH-indol-3-
I mL of 0.1 M sodium hydroxide is equivalent to 41.58 mg yl]acetyl]oxy]acetyl]oxy]acetic acid.
of C 21H1SCINO•. _____________________ '''''11
STORAGE
IMPURITIES
Specified impurities A, B, C,' D, E, F.
co,H
P
Cl
A. 4-cWorobenzoic acid,
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I-56 Acenocoumarol 2022
MOBILE PHASE
Acenocoumarol 20 volwnes of glacial acetic acid, 50 volumesof cyclohexane
and 50 volumes of dichtoromethane.
LIMITS
Any secondary spot in the chromatogram obtained with
solution (1) is not more intense thanthe spot in the
chromatogram obtained with solution (2) (0.1 %).
Loss on drying
When dried to constant weight at 105°) loses not more than
and enantiomer
0.5% of its weight. Use I g.
353.3 152-72-7 Sulfated ash

Not more than 0.1 %) Appendix IX A.
Action and use ASSAY
Vitamin K epoxide reductase inhibitor; oral anticoagulant. Dissolve0.6 g in 50 mL of acelOne and titrate with 0.1.'.1
Preparation sodium hydroxide VS using bromothymoJ blue solution R3 as
Acenocoumarol Tablets indicator. Repeatthe operation without the substance being
examined. The difference between the titrations represents
DEFINITION the amount of sodiumhydroxide required. Each mL of a.1M
Acenocoumarol is (RS)-4-hydroxy-3-(I-p-nitrophenyl-3- sodium hydroxide VS is equivalent to 35.33 mg of
oxoburyljcoumarin. It contains not less than98.5% and not CI9HlSN06'
more than 100.5% of C19HlSN06J calculated with reference
to the dried substance.
CHARACTERISTICS
An almost white to buff powder. Acesulfame Potassium
Practically insoluble in water and In ether, slightly soluble in
(ph. Bur. monograph 1282)
ethanol (96%). It dissolves in aqueous solutions of the alkali
hydroxides. It exhibits polymorphism.
IDENTIFICATION
The infrared absorption spellrnm, Appendix IT A, is concordant
with the reference spectrum of acenocoumarol (RS 001). If the
spectra are not concordant, dissolve 0.1 g of the substance
being examined in 10 mL of acetone and add water drop wise
until the solution becomes turbid. Heat on a water bath until
C,H,KNO,S 201.2 55589-62-3
the solution is clear and allow [0 stand. Filter, wash the
crystals with a mixture of equalvolumes of acetone and water Action and use
and dry at 100° at a pressure of 2 kPa for 30 minutes. Sweetening agent.
Prepare a new spectrum of the residue.
PhEw ~ _
TESTS
Clarity and colour of solution DEFINITION
A. A 2.0% w/v solution in cutlOne is dear) Appendix IV A. Potassium 6-methyl-I)2,3-oxathiazin-4-0Iate 2,2-dioxide.
B. The absorbanu of a 4-cm layer of a 2.0% w/v solution in Content
acelOne at 460 om is not more than 0.12) Appendix n B. 99.0 per cent to 101.0 per cent (dried substance).
C. A 2.0% wlv solution in O.IM sodium hydroxide is clear) CHARACTERS
Appendix N A, and yellow. Appearance
Light absorption White or almost white) crystalline powder or colourless
Absorbance of a 0.001 % wlv solution in a mixture of crystals.
I volume of 1M hydrochlori< acid and 9 volumes of methanol Solubility
at the maximum at 306 urn, 0.50 to 0.54, calculated with Solublein water) veryslightly soluble in acetone and in
reference to the dried substance) Appendix n B. ethanol (96 per cent).
Related substances IDENTIFICATION
Carry out the method for chin-layer chromalOgraphy,
First identification: A) C.
Appendix ill A) using the following solutions in acetone.
Second identification: B, C.
(1) 2.0% w/v of the substance beingexamined.
A. Infrared absorption spectrophotometry (2."2.24).
(2) 0.0020% wlv of the substance being examined.
Comparison acesu/fame potassium CRS.
CHROMATOGRAPHIC CONDITIONS
B. Thin-layer chromatography (2.2.27).
(a) Use as the coating silica gel GFZ54'
Test solution Dissolve 5 mg of the substance to be examined
(b) Use the mobile phase as described below. in water R and dilute to 5 mL with the same solvent.
(c) Apply 20 ~ of each solution. Reference solution (a) Dissolve 5 mg of acesulfame
(d) Develop the plate to 15 em. potassium CRS in wat~ R and dilute to 5 mL with the same
(e) After removal of the plate) allow it to dry in air and solvent.
immediately examine under ultraviolet light (254 nm).
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2022 Acesulfame Potassium I-57
Reference solution (b) Dissolve 5 mg of acesulfome Test solution Dissolve 0.100 g of the substance to be
potassium CRS and 5 mg of saccharin sodium R in water Rand examined in water R and dilute to 10.0 mL with the same
dilute to 5 mL with the same solvent. solvent.
Plate cellulose for chromatography R as the coating substance. Reference solution (aJ Dissolve 4.0 mg of acesulfame potassium
J.HobiJe phase concentrated ammonia R, acewne R, ethyl impurity B CRS in waterR and dilute to 100.0 mL with the
acetate R (10:60:60 VIVIV). same solvent. Dilute 1.0 mL of the solution to 200.0 mL
with water R.
Application 5 nl, as bands.
Developmem Twice over 2/3 of the plate. Reference solution (b) Dissolve 0.100 g of the substance to
be examined in reference solution (a) and dilute to 10.0 mL
Drying In a current of warm ale. with the same solution.
Detection Examine in ultraviolet light at 254 nm. Column:
System suitability Reference solution (b): - size: 1= 0.25 m, 0 = 4.6 mm;
- the chromatogram shows 2 clearly separated zones. - stationaryphase: octad«ylsi/yl silica gelfor chromatography R
Results The principal zone in the chromatogram obtained (3 urn).
with me test solution is similar in position and size to the J\1obile phase Mix 40 volumes of acetonitrile Rand
principal zone in the chromatogram obtained with reference 60 volumes of a 3.3 gIL solution of tetrabutylammollium
solution (a). hydrogen sulfateR.
C. 0.5 mL of solution S (see Tests) gives reaction (b) of Flow rate I mUmin.
potassium (2.3.1). Detection Spectrophotometer at 234 nm.
TESTS Injection 20 IJL.
Solution S Run time Twice the retention time of acesulfame.
Dissolve 10.0 g in carbon dioxide-free waur R and dilute to
Relative retention With reference to acesulfame (retention
=
time = about 5.3 min): impurity B about 1.6.
Appearance of solution
System suitability:
Solution S is clear (2.2.1) and colourless (2.2.2, Method If).
- signal-to-noise ratio: minimum 10 for the peak due to
Acidity or alkalinity impurity B in the chromatogram obtained with reference
To 20 mL of solution S add 0.1 mL of bromothymol blue solution (a);
solution RI. Not more than 0.2 mL of 0.01 M hydrochloric - peak-toJUal/ey ratio: minimum 1.2, where Hp =height
acid or 0.01 AI sodium hydroxide is required to change the above the baseline of the peak due to impurity Band
colour of the indicator." H; = height above the baseline of the lowest point of the
Impurity A curve separating this peak from the peak due to
Thin-layer chromatography (2.2.27). acesulfame, in the chromatogram obtained with reference
solution (b).
Test solutwn Dissolve 0.80 g of the substance to be
examined in water R and dilute to 10 mL with the same Limit:
solvent. - impmity B: not more than the area of the principal peak in
Reference solution (aJ Dissolve 50 mg of acetylacetamide R the chromatogram obtained with reference solution (a)
(impurity A) io water R and dilute to 25 mL with the same (20 ppm).
solvent. To 5 mL of the solution add 45 mL of water Rand Fluorides
dilute to 100 mL with methanolR. Maximwn 3 ppm.
Reference solution (bJ To 10 mL of reference solution (a) Potentiometry (2.2.36, Method f).
add 1 mL of the test solution and dilute to 20 mL with Test solution Dissolve 3.000 g of the substance to be
methanol R. examined in distilled waterR, add 15.0 mL of total-ionic-
Plate TLC silica gd plate R. strength-adjustment bufferR 1 and dilute to 50.0 mL with
Afobile phase waterR, ethanol (96 per cent) R, ethyl acetate R distiHed water R.
(2:15:74 VIVIV). Reference solutions To 0.5 mL, 1.0 mL, 1.5 mL and 3.0 mL
Application 5 ~L. of fluoride standard solution (10 ppm F) R add 15.0 mL of
total-ionic-strength-adjustment buffer RI and dilute to 50.0 mL
Development Over 2/3 of the plate.
with distilled warerR.
Drying In air until the solvents are completely removed.
Indicatorelectrode Fluoride-selective.
Detection Spray with phosphoric vanillin solution R and heat
Reference electrode Silver-silver chloride.
at 120 °C for about 10 min; examine in daylight.
System suiUlbility The chromatogram obtained with
Maximum 1.0 per cent, determined on 1.000 g by drying io
reference solution (a) shows a clearly visible spot and the
an oven at 105 °C for 3 h.
chromatogram obtained with reference solution (b) shows
2 clearly separated spots. ASSAY
Limit: Dissolve 0.150 g io 50 mL of anhydrous acetic acid R. Titrate
- impw;ty A: any spot due to impurity A is not more with 0.1 kI perchlotic add, determining the end-point-
intense than the spot in the chromatogram obtained with potentiometrically (2.2.20).
reference solution (a) (0.125 per cent). I mL of 0.1 M perchlaric acid is equivalent to 20.12 mg
Impurity B of C.H.,KNO.S.
Liquid chromatography (2.2.29). IMPURITIES
Specified impurities A, B.
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I-58 Acetazolamide 2022
H'CyO B. Infrared absorption spectrophotometry (2.2.24).

'yNH 2 Comparison acetazolamide CRS.
If the spectra obtained in the solid stateshowdifferences,
o
dissolve the substance to be examined and the reference
A. 3-oxobutanamide (acetylacetamide), substance separately in ethanol (96 per cent) R. evaporate to
dryness and record new spectra usingthe residues.
C. Introduce about 20 mg into a test-tube andadd 4 mL of
dilute hydrochloric acid Rand 0.2 g of zinc powder R.
Immediately place a piece of lead acetate paper R overthe
mouth of the tube. The paper shows a brownish-black
colour.
B. 5-<:h1oro-6-methyl-I,2,3-oxathiazin-4(3H)-one 2,2-<1ioxide. D. Dissolve about 25 mg in a mixture of 0.1 mL of dIlute
____ ~ PhE" sodium hydroxide solution Rand 5 mL of water R. Add 0.1 mL
of copper sulfate solution R. A greenish-blue precipitate is
formed.
TESTS
Acetazolamide Appearance of solution
The solution is not more opalescent than reference
(Ph Bur. monograph 0454) suspension II (2.2.1) and not more intensely coloured than
reference solution Yj or BYs (2.2.2, Method II).
Dissolve 1.0 g in 10 mL of 1 M sodium hydroxide.
Related substances
Test sdution Dissolve 40 mg of the substance to be
222.2 59-66-5
examined in the mobilephase and dilute to 100.0 mLwith
Action and use the mobile phase.
Carbonic anhydrase inhibitor; diuretic; treatment of Reference solution (a) Dilute 1.0 mL of the test solution to
glaucoma and ocular hypertension; treatment of mountain 100.0 mL with the mobile phase. Dilute 1.0 mL of this
sickness. solution to 10.0 mL with the mobile phase.
Preparation Reference solution (b) Dissolve the contents of a vialof
Acetazolamide Tablets acetazolamide for system suitability CRS (containing
impurities A, B, C, D, E and F) in 1.0 mL of the mobile
PhE" _
phase.
DEFINITION Column:
N-(5-Sulfamoyl-I,3,4-thiadiazol-2-yl)acetamide. - size: I =0.1 5 m, 0 =4.6 mm;
- stalWnary phase: end-capped praPoxybenzene SIlica gelfor
Content
chromalOgraphy R (4 pm).
Mobile phase acewnitrile for chromawgraphy R, 6.8 gIL
CHARACTERS solution of potassium dihydrogen phosphate R (10:90 VII').
Appearance
FWw rale 1.0 mllmin.
Whiteor almost white,crystalline powder.
Solubility
Very slightly soluble in water, slightly soluble in ethanol
Injection 25 ~l.
(96 per cent). It dissolves in dilute solutions of alkali Run lime 3.5 times the retention time of acetazolamide.
hydroxides. Identification of impumies Use the chromatogram supplied
It shows polymorphism (5.9). with acetazolamide for system suitability CRS and the
chromatogram obtained with reference solution (b) to
IDENTIFICATION identify the peaks due to impurities A, B, C, D, E and F.
First identification: A, B.
Relative retention With reference (Q acetazolamide (retention
Second identification: A, C~ D. time=about 8 min): impurity E =about 0.3;
A. Ultraviolet and visible absorption spectrophotometry impurity D = about 0.4; impurity B =about 0.6;
(2.2.25). impurity C =about 1.4; impurity A =about 2.1;
Solutum A Dissolve 30.0 mg in 0.01 M sodium hydroxide and impurity F =about 2.6.
dilute to 100.0 mL with the same solvent. Dilute 10.0 mL of System suitability Reference solution (b):
the solution to 100.0 mL with 0.01 M sodium hydroxide. - resolution: minimwn 2.0 between the peaks due to
Solution B Dilute 25.0 mL of solution A to 100.0 mL with impurities E and D.
0.01 M sodium hydroxide. Limits:
Spectral range 230-260 nm for solution A; 260-350 nm for - CQ17'UUon factors: for the calculation of content, multiply
solution B. the peak areas of the following impurities by the
Absorption maximum At 240 run for solution A; at 292 run corresponding correction factor: Impurity B = 2.3;
for solution B. impurity C = 2.6; impuriry D = 1.6;
- impurities A, B, C, D, E, F: for each impurity, not more
Specific absorbance at the absorption maximum 162 to 176 for
than 1.5 times the area of the principal peak in the
solution A; 570 to 620 for solution B.
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2022 Acetic Acid I-59
chromatogram obtained with reference solution (a)

(0.15 per cent);
- unspecified impurities: for each impurity, not more than the
with reference solution (a) (0.10 per cent); E. 5-acetamido-I,3,4-thiadiazole-2-sulfonic acid,
(0.6 per cent);
the chromatogram obtained with reference solution (3)
(0.05 per cent). F. N-[5-[(5-acetamido-I,3,4-thiadiazol-2-yl)
Sulfates (2.4.13) sulfonyl] sulfamoyl-l,3,4-thiadiazol-2-yl]acetamide,
Maximum 500 ppm.
To 0.4 g add 20 mL of distilled warer R and dissolve by
heating to boiling, Allow to cool with frequent shaking and
filter.
Loss on drying (2.2.32) G. 5-amino-I,3,4-thiadiazole-2-thiol.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
an oven at 105 DC.
ASSAY
Glacial Acetic Acid
Dissolve 0.200 g in 25 mL of dimethylformamide R. Titrate (Ph. Eur. monograph 0590)
with 0.1 M ethanolic sodium hydroxide, determining the
1 mL of 0.1 kI emanolic sodium hydroxide is equivalent to
22.22 mg of C.HoN,03S2'
IMPURITffiS
c,H,O, 60.1 64-19-7
Specified impurities AJ .8J CJ DJ EJ F. PhE" _
Otherdetectable impurities (thefollowing substances would, if
present ar a sufficient levelJ be detected by one or other of the tests DEFINITION
in the monograph. They are limited by thegeneral aueprance Content
criterion for other/unspecified impun"ries and/or by the general 99.0 per cent m/m to 100.5 per cent m/m.
monograph Substances for pharmaceutical use (2034). It is CHARACTERS
therefore nor necessary to identify these impun'ries for Appearance
demonstration of compliance. See also 5. JO. Control 0/impurities Crystalline mass or clear, colourless, volatile liquid.
in substances for pharmaceutical use) G.
Solubility
Miscible with water, with ethanol (96 per cent) and with
methylene chloride.
IDENTIFICATION
A. A 100 gIL solution is strongly acid (2.2.4).
A. N-(S-ehloro-lJ3A-thiadiazol-2-yl)acetamide, B. To 0.03 mL add 3 mL of warer R and neutralise with
dihue sodium hydroxide solution R. The solution gives
reaction (b) of acetates (2.3.1).
TESTS
Solution S·
B. N-(1,3,4-thiadiazol-2-yl)acetamide, Dilute 20 mL to 100 mL with disrined waterR.
Appearance
The substance to be examined is clear (2.2.1) and colourless
(2.2.2, Method11).
Freezing point (2.2. II!)
Minimum 14.8 -c,
C. N-(5-sulfanyl-I,3,4-thiadiazol-2-yl)acetamide,
Reducing substances
Dilute 2.0 mL to 10.0 mL with warer R. Add 0.1 mL of
0.02 M potassium permanganate. Heat on a water-barn. for
1 min, the colourremains pink.
Chlorides (2.4.4)
Maximum 25 mgIL.
D.S-amino-l,3,4-thiadiazole-2-sulfonamide,
Dilute 10 mL of solution S to 15 mL with water R.
Sulfates (2.4.13)
Maximum 50 mg/L, determined on solution S"
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1-60 Acetic Acid 2022
Iron (2.4.9) Readily oxldisable Impnritles

Maximum 5 ppm. To 25 mL add 0.2 mL of 0.02M potassium pennanganate VS
Dissolve the residue obtained in the test for residue on and allow to standfor 1 minute. The pink colour is not
evaporation by heating with 2 quantities, each of 15 ml., of entirely discharged.
water R and dilute to 50.0 mL with water R. Dilute 5.0 mL Non-volatile matter
of the solution to 10.0 mL with wattr R. When evaporated {Q dryness and dried at 105°, leaves not
Residue on evaporation more than 0.01% wlw of residue.
Maximum 0.01 per cent. ASSAY
Evaporate 20 g to dryness on a water-bath and dry at Add 30 mL of water to 20 g in a stopper flask and titrate
100-105 °C. The residue weighs a maximum of 2.0 mg. with 1M sodium hydroxide VS using phenolphthalein solution Rl
ASSAY as indicator. Each rnL of 1M sodium hydroxide VS is
Weigh accurately a conical flask with a ground-glass stopper '0
equivalent 60.05 mg of C,H.O,.
containing 25 mL of water R. Add 1.0 mL of the substance
to be examined and weigh again accurately. Add 0.5 mL of
phenolphthalein solution R and titrate with 1 M sodium
hydroxide. Acetic Acid (33 per cent)
"I mL of 1 M sodium hydroxide is equivalent to 60.1 mg Acetic Acid
ofC,H,O,. Preparation
STORAGE Acetic Acid (6 per cent)
In an airtight container. DEFINITION
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE« Acetic Acid (33 per cent) contains not less than 32.5% and
not more than 33.5% w/w of acetic acid, ~Ht02'
CHARACTERISTICS
A clear,colourless liquid.
Acetic Acid (6 per cent) Miscible with water, with ethanol (96%) and with glycerol.
Dilute Acetic Acid
IDENTIFICATION
DEFINITION A. Strongly acidic, even when dilutedfreely.
Acetic Acid (6 per cent) containsnot less than 5.7% and not B. When neutralised, yields the reactions characteristic of
more than 6.3% w/w of acetic acid, ~H.t02' It may be aalates, Appendix VI.
prepared by mixing 182 g of Acetic Acid (33 per cent) with
818 g of Purified Wa'er. TESTS
Weight per mL
IDENTIFICATION
1.040 10 1.042 g, Appendix V G.
A. Strongly acidic.
Chloride
B. When neutralised, yields the reactions characteristic of
Dilute 5.0 mL with sufficient water 10 produce 100 mL.
acetates, Appendix VI.
15 mL of the resulting solutioncomplies with the limit test for
TESTS chlorides, Appendix VII (70 ppm).
WeIght per mL Sulfate
About 1.005 g, Appendix V G. 12.5 mL of the solutionused in the test for Chloride, diluted
Chloride to 15 mL with water, complieswith the limitces, for sulfates,
Dilute 5.0 mL with sufficient water to produce 100 mL. Appendix VII (240 ppm).
15 mL of the resulting solution complies with the limit teu for Aldehydes
chlorides, Appendix VII (70 ppm). Distil 15 mL. To the first 5 mL of the distillate add 10 mL
Sulfate of a 5% wlv solution of mercury(JI) chloride, make alkaline
12.5 mL of the solution used in the test for Chloride, diluted with 5M sodium hydroxide, allow to stand for 5 minutes and
lO 15 mL with Water, complies with the limit test for suifateS, make acidic with 1M suJfun'c add. The solutionshows not
Appendix VII (240 ppm). more than a faint turbidity.
Aldehydes Formic acid and oxidisable impurities
Distil 75 mL. To the firs' 5 mL of the distillate add 10 mL '0
Mix 5 mL with 6 mL of sulfuric acid and cool 20'.
of a 5% wlv solution of mercury(ll) chloride, makealkaline Add 2 mL ofO.0167M potassium dichromate VS, allow '0
with 5M sodium hydroxide, allow to stand for 5 minutes and stand for I minute, add 25 mL of water and I mL of freshly
acidify with 1Msulfuric acid. The solution shows not more prepared dilute potassium iodide solution and titrate the
than a faint turbidity. liberated iodine with O.IM sodium thiosulfate VS using starch
Formic acId and oxldisable Impnritles mucilage as indicator. Not less than 1.0 mL of O.lM sodium
'0
Mix 5 mL with 6 mL of su/juric acid and cool 20'. tlDosu!!ate VS is required.
Add 0.4 mL ofO.0167M potassium dichromate VS, allow '0 Readily oxldisable Impnritles
stand for I minute, add 25 mL of waterand I mL of freshly To 5.0 mL add 20 mL of water and 0.2 mL of
prepared diluu potassium iodide solution and titrate the 0.02M potassium permanganare VS and allow to stand for
liberated iodine with O.IM sodium thiosulfate VS using starch 1 minute. The pink colour is not entirely discharged.
mucilage as indicator. Not less than 0.2 mL of O.IM sodium Non-volatile matter
thiosulfate VS is required. When evaporated to dryness and driedat 105°, leavesnot
more than 0.01% w/w of residue.
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2022 Acetone 1-61
ASSAY Reference solution (b) Dilute 100 ilL of benzene R to

Weigh 5 g into a stopperflask containing 50 mL of water and 100.0 mL with the test solution. Dilute 0.20 mL of this
titrate with 1Msodium hydroxide VS using phenolphthalein solution to 100.0 mL with the test solution.
sohuion RJ as indicator. Each mL of 1M sodium hydroxide VS Column:
is equivalent to 60.05 mg of C2H4 0 2 . - material: fused silica,
- size: / ;;;; 50 rn, 0 ;;;; 0.3 mm,
- stationary phase: mtu:rogof 20 000 R (film thickness 1 urn).
Carrier gas helium for chromatography R.
Acetone ***
** ** Linearvelocity 21 crn/s,
(ph. Bur. monograph 0872) ***** Spill ratio 1:50.
Temperature:
Time Temperature
(mlo) CCJ
Column 0- Il 45 ---> 100
C,H.O 58.08 67-64-1 II - 20 '00
PhEu ~ _ Injection port 150
Detector 250
DEFINITION
Propanone. Detection Flame ionisation.
CHARACTERS Injeaion I ~L.
Appearance Retention time Impurity C ;;;; about 7.5 min.
Volatile, clear, colourless liquid.
System suitability.
Solubility - resolution: minimum 5.0 between the peak due to
Miscible with water and with ethanol (96 per cent). impurity A (2nd peak) and the peak due to impurity B
The vapour is flammable. (3rd peak) in the chromatogram obtainedwith reference
IDENTIFICATION solution (a);
- signal-to-noise ratio: minimum 5 for the peak due to
A. Relative density (see Tests).
impurity C in the chromatogram obtained with reference
B. To I mL, add 3 mL of difute sodium hydroxide sofution R solution (b).
and 0.3 mL of a 25 gIL solution of sodium nitropmsside R.
Limits:
An intensered colour is produced which becomes violet with
- impurities A, B: for each impurity, not more than the
the addition of 3.5 mL of acetic acid R.
difference between the areas of the corresponding peaks in
C. To 10 mL of a 0.1 per cent VIV solutionof the substance the chromatogram obtained with reference solution (a)
to be exantined in ethanol(50 per cent V/~ R, add I mL of a and the areas of the corresponding peaks in the
10 gIL solution of nitrobenzafdehyde R in ..hanol chromatogram obtained with the test solution
(50 per cent V/~ R and 0.5 mL of strong sodium hydroxide (0.05 per cent VIII),
solution R. Allow to stand for about 2 min and acidify with - impuniy C: not more than me difference between the area
acetic acid R. A greenish-blue colouris produced. of me peak due to impurity C in the chromatogram
TESTS obtained with reference solution (b) and the area of the
Appearance of solution corresponding peak in the chromatogram obtained wirh
To 10 mL add 10 mL of waterR. The solution is clear the 'est solution (2 ppm VIII),
(2.2.1) and colourless (2.2.2, MelhodII). - any other impun'ty: for each impurity, not more than the
difference between the area of the peak due to impurity A
Acidity or alkalinity
To 5 mL add 5 mL of carbon dioxide-free waterR, 0.15 mL of
and the area of the corresponding peak in the
phenolphlhalein solution Rand 0.5 mL of O. 01 M sodium
chromatogram obtained with the test solution
hydroxide. The solution is pink. Add 0.7 mL of 0.01 M
(0.05 per cent VIII).
hydrochloric acid and 0.05 mL of methyl redsolution R.
The solution is red or orange. Matter insoluble in water
Dilute 1.0 mL to 20 mL with water R. The solution is clear
Relative density (2.2.5)
(2.2.1).
0.790 to 0.793.
Residue on evaporation
Reducing substances
Maximum 50 ppm.
To 30 mL add 0.1 mL of 0.02 M potassium permanganate
and allow to stand in the dark for 2 h. The mixture is not Evaporate 20.0 g to dryness on a water-bath and dry at
completely decolourised. 100-105 DC. The residue weighs a maximum of 1 mg.
Related substances Water (2.5.12)
Gas chromatography (2.2.28). Maximum 3 gIL, determined on 10.0 mL.
Testsolutkm The substance to be examined. STORAGE
Reference solution (a) To 0.5 mL of melhanof R add 0.5 mL Protected from light.
of 2-propanol R and dilute to 100.0 mL with the test solution. IMPURITIES
Dilute 1.0 mL of this solution to 10.0 mL with the test Specified impurities A, B, C.
solution.
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........ -
1-62 Acetylcholine Chloride 2022
TESTS
Solution S
A. methanol, Dissolve 5.0 g in carbon dioxide-free water R and dilute to
Solution S is clear (2.2.1) and not more intensely coloured
than reference solution Y. or BY. (2.2.2, MeJhod If).
B. propan-z-ol (isopropanol), Acidity
o
Dilute 1 mL of solution S to 10 mL with carbon dioxide-free
water R. Add 0.05 mL of phenolphthalein solution R. Not more
than 0.4 mL of 0.01 M sodium hydroxitk is required to
changethe colour of me indicator to pink.
c. benzene. Related substances
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'I Thin-layer chromatography (2.2.27). Prepare the solutions
immediately before use.
Test solution (a) Dissolve 0.30 g of the substance to be
examined in methanol R and dilute to 3.0 mLwith the same
Acetylcholine Chloride solvent.
Test solution (b) Dilute I mL of test solution (a) to 10 mL
(Ph. Bur. monograph 1485) with methanol R.
Reference solution (a) Dilute I mL of test solution (a) to
100 mL with methanol R.
Reference solution (b) Dissolve 20.0 mg of acetylcholine
chloride CRS in methanol R and dilute to 2.0 mL with the
181.7 60-31-1 same solvent.
Reference solution (c) Dissolve 20 mg of choline chloride R in
Action and use
methanol R, add 0.4 mL of test solution (a) anddilute to
Cholinoceptor agonist.
2.0 mL with methanol R.
PhE<I _
Plate TLC silica gelplateR.
DEFINITION Mobile phase Mix 20 volumes of a 40 gIL solution of
2-(Acetyloxy)-N,N,N-trimethylethanaminium chlorlde. ammonium nitrate R, 20 volumes of methanolRand
60 volumes of acetonitrile R.
Content
98.5 per cent to 101.5 per cent (dried substance). Application 5 Il1. as bands of 10 mm by 2 mm.
Deve/opmenl Over 2/3 of the plate.
CHARACTERS
Appearance
Detection Spraywith potassium iodobismuthat6 solution R3.
White or almost white crystalline powderor colourless System suitability The chromatogram obtained with
crystals, very hygroscopic. reference solution (c) shows 2 clearly separated zones.
Solubility Limits:
Verysoluble in water, freely soluble in alcohol, slightly - any impun"ry: any zones in the chromatogram obtained
soluble in methylene chloride. with test solution (a), apart from the principal zone, are
not more intense than the principal zone in the
IDENTIFICATION chromatogram obtainedwith reference solution (a)
First identification: B, E. (1 per cent).
Second identification: A, C, D, E. Trimethylamine
A. Melting point (2.2.1f): 149 "C to 152 "C. Dissolve 0.1 gin 10 mL of sodium carbonate solution R and
Introduce the substance to be examined into a capillary tube. heat to boiling; No vapours appear which rum red litmus
Dry in an oven at 100-105 "C for 3 h. Seal the tube and paper R blue.
determine the melting point. Loss on drying (2.2.32)
B. Infrared absorption spectrophotometry (2.2.2f). Maximum 1.0 per cent, determined on 1.000 g by drying in
Comparison atetylcholine chloride CRS. an oven at 105 °C for 3 h.
C. Examine the chromatograms obtained in the test for Sulfated ash (2.4.1f)
related substances. Maximum 0.1 per cent, determined on the residue obtained
Results The principal zone In the chromatogram obtained in the test for loss on drying.
with test solution (b) is similar in position, colour and size to ASSAY
the principal zone in the chromatogram obtained with Dissolve 0.200 g in 20 mL of carbon dioxide-free water R.
reference solution (b). Neutralise with 0.01 M sodium hydroxide using 0.15 mL of
D. To 15 mg add 10 mL of dilute sodium hydroxide solution R, phenolphthalein solution R as indicator. Add 20.0 mL of 0.1 M
2 mL of 0.02 M potassium permanganate and heat. sodium hydroxide and allow to stand for 30 min. Titratewith
The vapours formed change the colourof red litmus paper R 0.1 M hydrochloris acid.
to blue. I mL of 0.1 M sodium hydroxide is equivalent to 18.17 mg of
E. 0.5 mL of solution S (see Tests) gives reaction (a) of G,H,.ClN02 •
chlorides (2.3.1).
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2022 Acerylcysteine 1-63
STORAGE Determination B Mix equal parts of the substance to be

In ampoules, protected from light. examinedand acetylcysteine CRS and determine the melting
point of the mixture.
IMPURITIES
Result B The absolutedifference between the meltingpoint
of the mixture and the valueobtained in determination A is
not greater than 2 "C.
C. Infrared absorption spectrophotometry (2.2.24).
A. 2-hydroxy-N,N,N-trimethylethanaminium chloride Comparison aeety/cysteine CRS.
(choline chloride), TESTS
The solution is clear (2.2.1) and colourless (2.2.2,
Method If).
Dissolve 0.5 g in waterR and dilute to 10 mL with the same
B. 2-(acetyloxy)-N,N-dimethylethanaminium chloride, solvent.
Specific optical rotation (2.2.7)
+ 21.0 to + 27.0 (dried substance).
Mix 1.25 g and I mL of a 10 gIL solution of sodium
edeuue R. Add 7.5 mL of a 40 gIL solution of sodium
C. N,N-dimethylmethanamine. hydroxide R, mix and dissolve. Dilute to 25.0 mL with
____________________ ""E" phosphale buffer solution pH 7.0 R2.
Related substances
Liquid chromatography (2.2.29). Prepare the solutions
Acetylcysteine Solution A 1.03 gIL solution of hydrochlori< acid R.
Testsolution Suspend 0.120 g of the substance to be
(Ph. Bur. monograph 0967) examined in solution A and dilute to 15.0 mL with
solution A, ensuring complete dissolution.
50.0 mL with solution A. Dilute 1.0 mL of this solution to
100.0 mL with solution A.
Reference solution (b) Dissolve 4 mg of L-cystine R
(impurity A) in solution A and dilute to 10 mL with
C,H,NO,S 163.2 616-91-1 solution A.
Reference solution (c) Dissolve 3 mg of l-cysteine R
Action and use
(impurity B), 5 mg of acetylcysteine impurity C CRS and
Sulfydryl donor; antidote to paracetamol poisoning;
2.5 mg of tu:etykysteine impun'ty D CRS in solution A, mix
mucolytic.
with 4 mL of reference solution (b) and dilute to 20 mL with
Preparations solution A. Dilute I mL of this solution to 10 mL with the
Acetylcysteine Eye Drops test solution.
Acetylcysteine Injection Reference solution (d) Dissolve 2 mg of sodium 2-merhyl-2-
""E" _ thiazoline-4-carboxylate R in solution A and dilute to 50 mL
with solution A.
DEFINITION Column:
(2R)-2-Acetamido-3-sulfanylpropanoic acid. - size: I::::: 0.25 m, 0 ::::: 4.0 mm;
Content - stationary phase: end-eapped tutadecylsilyl silica gelfor
98.5 per cent to 101.0 per cent (dried substance). chromatography R (5 urn).
CHARACTERS jHobiJe phase acetonitrile for chromatography RJ waterfor
Appearance chromatography R previously adjusted to pH 3.0 with
White or almost white, crystalline powderor colourless phosphoric acidR (3:97 VW).
crystals. Flow rale 1.0 mllmin.
Solubility Detection Spectrophotometer at 220 om.
Freely soluble in water and in ethanol (96 per cent), Injection 20 ilL of the test solutionand reference
practically insoluble in methylene chloride. solutions (a), (c) and (d).
IDENTIFICATION Run time 3 times the rerention time of acetylcysteine,
First identification: A, C. Identification of impurities Use the chromatogram obtained
Second idetuificauon: B. with reference solution (c) to identify the peaks due to
impurities AJ BJ C and D; use the chromatogram obtained
A. Specific optical rotation (see Tests).
with reference solution (d) to identify the peak due to
B. Melting point (2.2.14). 2-methyl-2-thiazoline-4-carboxylic acid.
Determination A Determine the melting point of the Relative retention With reference to acetylcysteine (retention
substance to be examined. time = about 5 min): impurity A = about 0.48;
Result A 108"C to IlO "C.
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1-64 Acetylcysteine 2022
impurity B = about 0.53; 2-methyl-2-thiazoline-4-carboxylic STORAGE

acid = about 0.8; impurity C = about 2.1; Protectedfrom light.
impurity D = about 2.6.
IMPURITIES
Systemsuitability: Specified impurities B, C, D.
- resolution: minimwn 1.5 between me peaks due to
Other detectable impurities (the joUowing subsrances would, if
impurities A and B in the chromatogram obtained with
present at a suJficienc level, be detected by one or other of the tests
reference solution (c)j
in the monograph. They are limited by thegeneral a«eptance
- peak-w-1Jalley ratio: minimwn 5.0, where Hp = height
criterion for other/unspecified imPJm·ties. It is therefore not
above the baselineof the peak due to 2-methyl-2-
necessary UJ identify these impurities for demonstration of
thiazoline-4-carboxyHc acid and H; = height above the
compliance. See also 5.10. Control of~'mpuniies in substances/or
baselineof the lowest point of the curve separating this
pharmaceutical use) A.
peakfrom the peak due to acerylcysteine in the
chromatogram obtained with reference solution (e);
- symmetry factor. maximum 2.2 for the peak due to
acetylcysteine in the chromatogram obtained with
reference solution (a).
Calculation of percentage contents:
- correction factors: multiply the peak areas of the following A. 3,3'-disulfanediylbis[(2R)-2-aminopropanoic acid] (L-
impurities by the corresponding correction factor: cystine),
impurity B = 3.4; impurity C = 0.7; impurity D = 0.3;
- for each impurity) use the concentration of acetylcysteine H Nfl,
in reference solution (a). HS0 CO,H
Limits:
- impun'ty C: maximum 0.3 per cent, B. (2R)-2-amino-3-sulfanylpropanoic acid (t-cysteine),
- impurity B: maximum 0.2 per cent;
- impun"ty D: maximum 0.15 per cent;
- unspecified ;mpun"ties: for each impurity) maximum
0.10 per cent;
- total: maximum 0.5 per cent;
- reporting threshold: 0.05 per cent; disregard the peakdue to
2-methyl-2-thiazoline-4-carboxylic acid, which is formed
due to in situ degradation of acetylcysteine in acidic
solutions such as solution A.
The thresholds indicated under Related substances C. 3,3'-disulfanediylbis[(2R)-2-acetamidopropanoic acid] (N,
(Table 2034.-1) in the general monograph Substances for N'-diacetyl-L-cystine)J
pharmaceutical use (2034) do not apply.
Zinc
Maximum 10 ppm.
Atomic absorption spectrometry (2.2.23, Method If).
TestsolutUm Dissolve 1.00 g in a 0.103 gIL solution of
hydrochlori< acid R and dilute to 50.0 mL with the same
solution.
Reference solutions Prepare the reference solutions using zinc D. (2R)-2-acetamido-3-(acetylsulfanyl)propanoic acid (N,S-
standard solu.on (5 mglmL Zn) R, diluting with a 0.103 gIL diacetyl-L-cysteine).
solution of hydrochloric acid R. ________________ ~ PI>E"
Source Zinc hollow-cathode lamp.
Wavelength 213.9 nm.
Atomisation device Air-acetylene flame.
Use a correction procedure for non-specific absorption.
VlUUO at 70°C for 3 h.

ASSAY
Dissolve 0.140 g in 60 mL of water R and add 10 mL of
d/7uM hydrochloric acid R. Add 10 mL of porassium iodide
solution R and titrate with 0.05 kI iodine, determining the
1 mL of 0.05 M iodine is equivalent to 16.32 mg of
C,H,NO,S.
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2022 Acetyldigoxin 1-65
Reference solution (c) Dissolve 5 mg of gisoxin CRS

Acetyldigoxin (impurity D) in the solvent mixture and dilute to 100.0 mL
(p-Acety/digoxin, Ph. Eur. monograph 2168) with the solvent mixture. To 5.0 mL of this solution, add
0.5 mL of reference solution (a) and dilure to 100.0 mL with
o the solvent mixture.
Reference solution (d) Dissolve 5.0 mg of P-a<ety/digoxinfor
peak identification CRS (containing impurities A and B) in
10.0 mL of the solvent mixture.
Column:
- size: 1= 0.125 m,·0 = 4.0 mm;
- stationary phase: oetade<y/si/yl silica gelfor chromatography R
(4 pm).
Mobile phase:
- mobile phaseA: waterfor chromatography R;
- mobile phase B: acetonitrile for chromatography R;

(mIn) (per cent VIV) (per cent ViJ')
0-10 70 30
10 - 20 70 ..... 35 30 ..... 65
20·20.t 35 ..... 70 65 ..... 30
823 5355-48-6 20.1 - 25 70 30
Action and use

Cardiac Glycoside.
PhE" ~ _
Injection 10 r.tL of the test solution and reference
DEFINITION solutions (b), (c) and (d).
3 p-[(4- O-Acetyl-2,6-dideoxy-p-D-ribo-hexopyranosyl-(I ~ 4)- Identification of impurities Use the chromatograms obtained
2,6-dideoxy-p-D-ribo-hexopyranosyl-(1->4)-2,6-dideoxy-p-D- with reference solutions (c) and (d) to identify the peaks due
ribo-hexopyranosyl) oxy]-12P, 14-dihydroxy-Sjl-card-20(22)- to impurities A, Band D.
enolide. Relative retention With reference to p-acetyldigoxin
Content (retention time = about 9 min): impurity B' = about 0.3;
97.0 per cent to 102.0 per cent (dried substance). impurity A = about 0.7; impurity D = about 1.2.
CHARACTERS System suitability Reference solution (c):
Appearance - resolution: minimum 1.5 between the peaks due to
White or almost .white powder. IJ-acetyldigoxin and impurity D;
- symmetry factor: maximum 2.5 for the peak due to
Solubility Il-acetyldigoxin.
Practicajly insoluble in water, sparingly soluble in methylene
Limits:
chloride, slightly soluble in ethanol (96 per cent).
- impun'lies A B: for each impurity, not more than the area
j
IDENTIFICATION of the principal peak in the chromatogram obtained with

Infrared absorption spectrophotometry (2.2.24). reference solution (b) (0.5 per cent);
Comparison P-a<etyldigoxin CRS. - impun'ty D: not more than 0.6 times the area of the
principal peak in the chromatogram obtained with
TESTS
reference solution (b) (0.3 per cent);
- arry other impurity: for each impurity, not more than
+ 26.2 to + 28.2 (dried substance). 0.4 times the area of the principal peak in the
Dissolve 0.50 g in a mixture of equal volumes of methanol R chromatogram obtained with reference solution (b)
and methylene chloride R and dilute to 25.0 mL with the same (0.2 per cent);
mixture of solvents. - sum of impurities other than A Band D: not more than
j
Related substances 1.2 times the area of the principal peak in the
Liquid chromatography (2.2.29). Prepare the solutions chromatogram obtained with reference solution (b)
immediately before use. (0.6 per cent);
So/vent mixture Mix equal volumes of methanol R2 and - total: not more than 3 times the area of me principal peak
acetonitrile for chromatography R. in the chromatogram obtained with reference solution (b)
(1.5 per cent);
Testsolutio" Dissolve 50.0 mg of the substance to be
- disregard limit. 0.1 times the area of the principal peak in
examined in the solvent mixture and dilute to 100.0 mL with
the chromatogram obtained with reference solution (b)
the solvent mixture.
(0.05 per cent).
Reference solution (a) Dissolve 10.0 mg of
The thresholds indicated under Related substances
p-acetyldigoxin CRS in the solvent mixture and dllute to
(Table 2034.-1) in the general monograph Substances for
Reference solution (b) Dilute 1.0 mL of the test solution [0
20.0 mL with the solvent mixture. Dilute 1.0 mL of this
solution to 10.0 mL with the solvent mixture.
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1-66 Acetyldigoxin 2022

o
an oven at 105 "C.
Sulfated ash (2.4. /4)
Maximum 0.1 per cent, determined on me residue obtained
in the test for loss on drying.
ASSAY
related substances with the following modification. C. 3P, 12p, 14-trihydroxy-5p-card-20(22)-enolide
lnjection Test solution and reference solution(a). (digoxigenin),
Calculate the percentage contentof C43H66015 from the
declared content of p-a<:elyldigoxin CRS.
STORAGE
IMPURITIES
Specified impurities A, B, D.
Otherdetectable impuniies (thefollowing subseanas would, if
present at a sufficient level, be detected by one or other of the tests
in em monograph. They areUmited by the general aa:eptanu
oitenon for other/unspecified impuniies. It is therefore not
mxessary to identify these impurities for demonstration of
compliance. See also 5.10. Control ofimpun"ties in substances/or
pharmaceutical use) C, E, F, GJ H.
o o
D. 3jl-[(2,6-dideoxy-p-o-rib<>-hexopyranosyl-(l--> 4)-2,6-
dideoxy-p-o-rib<>-hexopyranosyl-(l-->4)-2,6-dideoxy-p-o-
rib<>-hexopyranosyI)oxy]-14, 16p-dihydroxy-5jl-card-20(22)-
enolide (gitoxin),
A. 3P-[(3-D-acetyl-2,6-dideoxy-p-o-ribo-hexopyranosyl-
(1-->4)-2,6-dideoxy-p-o-ribo-hexopyranosyHl--> 4)-2,6-
dideoxy-p-o-rib<>-hexopyranosyl)oxy)-12p, 14-<1ihydroxy-
5jl-card-20(22)-eno~de (c-acetyldigoxia),
E. 3P-[(2,6-dideoxy-p-o-rib<>-hexopyranosyl-(l-->4)-2,6-
dideoxy-jl-o-rib<>-hexopyranosyl-(l-->4)-2,6-dideoxy-p-o-
rib<>-hexopyranosyl) oxy)-14-hydroxy-5p-card-20(22)-
enolide (digitoxin),
FoP
H
~
CH30~ OH
OH
HO
OH
B. 3P-[(2,6-dideoxy-jl-o-rib<>-hexopyranosyl-(1 ~ 4)-2,6-
dideoxy-p-o-ribo-hexopyranosyl-(l-->4)-2,6-dideoxy-jl-o-
nb<>-hexopyranosyI)oxy]-12P, 14-dihydroxy-5jl-card-20(22)-
enolide (digoxin),
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2022 Acetylene Intermix (1 per cent) in Nitrogen 1-67
Acetylene Intermix (1 per cent) in

Nitrogen
(Ph. Enr. monograph 2903)
Ph,,, _
DEFINITION
A mixture containing I per cent VIV of acetylene in Low-
oxygen nitrogen (1685).
Content
0.95 per cent VIV to 1.05 per cent VIV of acetylene (C 2H2 )
in nitrogen (N2).
This monograph applies [0 acetylene intermix (l per cent) in
nitrogen used in the preparation of lung function test gas
mixtures for medicinal use.
PRODUCTION
F. 3P-[(3,4-0-diacetyl-2,6-dideoxy-p-D-n'b<>-hexopyranosyl- The acetylene used in the manufacturing process is produced
(I ~4)-2,6-dideoxy-p-D-ribo-hexopyrnnosyl-( I ~ 4)-2,6- by hydrolysis of calcium carbide.
dideoxy-p-D-n'bo-hexopyranosyl)oxy)-12P,14-dihydroxy- Prior [0 usingthe gas in the manufacturing process, the
5Il-card-20(22)-enolide (diacetyldigoxin), acetylene may be passed through an activated charcoal filter.
The acetylene is stored in cylinders, whichmay be filled with
a porous mass with acetone the only solvent permitted.
CHARACTERS
Appearance
Colourless gas.
IDENTIFICATION
A. Examine the chromatograms obtained in the assay.
Results The peak due to acetylene in the chromatogram
obtained with the gas to be examined is similar in retention
time and size to the peakdue to acetylene in the
chromatogram obtained with the reference gas.
B. Gas chromatography (2.2.28).
Gas to be examined The substance to be examined.
Reference gas Nitrogen RI.
Column:
- material: stainless steel;
G.31l-[(3-o-acetyl-2,6-dideoxy-p-D-ribo-hexopyrnnosyl- - size: / = 2 m, 0 = 2 mm;
(I ~4)-2,6-dideoxy-p-D-ribo-hexopyranosyl-(1 ~4)-2,6 - stationary phase: molecular sieve for chromatography R
dideoxy-p-D-ribo-hexopyrnnosyl) oxy]-I4-hydroxy-5Il-card- (0.5 nco).
20(22)-enolide (u-acetyldigitoxin), Carnergas helium for chromatography R.
a Flow rate 20 mUmin.
Temperature:
- column: 80°C;
- detector: 130 'C.
Detection Thermal conductivity.
Injection 10 ~L.
Reuntion time Nitrogen = about 2 min.
Results The principal peak in the chromatogram obtained
with the gas to be examined is similar in retention time to
the principal peak in the chromatogram obtained with the
reference gas.
TESTS
Acetone
Gas chromatography (2.2.28).
Reference gas Mixture containing 250 ppm VIV of ace"'", R
H. 3P-[(4-0-acetyl-2,6-dideoxy-p-D-ribo-hexopyrnnosyl- in nitrogen R.
(I ~4)-2,6-dideoxy-p-D-n'b<>-hexopyrnnosyl-( I ~ 4)-2,6-
Column:
dideoxy-p-D-ribo-hexopyrnnosyl) oxy]-14-hydroxy-5Il-card-
- material: fused silica;
20(22)-enolide (p-acetyldigitoxin).
___________________ Ph,,, - size: / = 10 ID, 0 = 0.53 IDID;
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1-68 Acetyltryptophan 2022
- stationary phase: poIyorganosiJoxane for oxygen-containing

compounds R (film thickness 10 urn).
Carrier gas heNum for chromatography R.
Flow rale 50 mUmin. B. arsane (arsine),
Temperature:
- column: 200°C;
- inj«tion port: 240 "C;
- detector: 250 "C.
Detection Flame ionisation. C. phosphane (phosphine),
Injection 25 ~L.
Retention time Acetone = about 1 min.
Calculation of percentage content:
- use the concentration of acetone in the reference gas. D. hydrogen sulfide,
Limit:
- amone: maximum 250 ppm VIV. H"O'H
•
Arsine
Maximum 0.25 ppm VIVo determined using an arsine E. water.
detector tube (2.1.6). _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE,;
Phosphine
Maximum 0.2 ppm VIV; determined using a phosphine
detector tube (2.1.6).
Hydrogen sulfide Acetyltryptophan
Maximwn 0.2 ppm ViVo determined using a hydrogen
sulfide detector tube (2.1.6). (N-AcetyltrypU>fJhan, Ph. Eur. monograph 1383)
Water (2.5.28)
Maximum 10 ppm VIV.
ASSAY
Gas chromatography (2.2.~8).
Reference gas Mixture containing 1.0 per cent VIV of C 13H,.,N,O, 246.3 87-32-1
acetylene R in nitrogen RI. PhE,; _
Column: DEFINITION
- material: stainless steel; (RSJ-2-Acetylamino-3-(IH-indol-3-yl)propanoic acid.
- size: 1= 2 m, 0 = 2 rnm;
- stationary phase: 3 per cent squalane R on alumina. Content
99.0 per cent to 101.0 per cent (driedsubstance).
Comer gas helium for chromatography R.
Flow rale 20 mUmin. PRODUCTION
Tryptophan used for the production of N-acetyltryptophan
Temperatu te :
- column: 100°C; complieswith the test for impurity A and otherrelated
substances in the monograph on TrypU>fJhan (1272).
- detector: 250 "C.
Detection Flame ionisation. CHARACTERS
Injection I 00 ~L. Appearance
White or almost white, crystalline powder, or colourless
Retention time Acetylene e about 6 min.
crystals.
Calculate the percentage content of C2H2 .
Solubility
STORAGE Slightly soluble in water, verysoluble in ethanol
As a compressed gas, in appropriate high-pressure cylinders (96 per cent). It dissolves in dilute solutions of alkali
complying with the legal regulations. hydroxides.
LABELLING mp
The label states the nominalcontent) in per cent VIV, of About 205 "C.
acetylene in nitrogen. IDENTIFICATION
IMPURITIES First idenu]icalion: A, B.
Specified ;mpun'ties A, B, C, D, E. Second identification: A, C, D, E.
A. Opticalrotation (see Tests).
B. Infrared absorption spectrophotometry (2.2.24).
Comparison N-acetyltryptophan CRS.
A. propan-2-one (acetone), Test solution Dissolve 50 mg of the substance to be
examined in 0.2 mL of concentrated ammonia R and dilute to
10 mL with water R.
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2022 Acetyltryptophan 1-69
Reference solution (a) Dissolve 50 mg of N- Time Mobile phase A Mobile phase B

acetyltryprophan CRS in 0.2 mL of concentrated ammonia R (min) (per cent VIJI) (per cent VIJI)
and dilute to 10 mL with water R. 0-10 '00 0
10 - 45 100 ...,. 0 0...,. 100
Reference solution (b) Dissolve 10 mg of tryptophan R in the
45·65 0 '00
test solution and dilute to 2 mL with the test solution.
PiaI' TLC silica gelFm piaI' R.
Flow raW 0.7 mIlmin.
J.Hobl1e phase glacial acetic add R, waterR, buumoiR
(25:25:40 VIVII'). Delation Spectrophotometer at 220 om.
Application 2 ~L. Injection 20 f.lL of the test solution and reference
solutions (a) and (c).
Development Over a path of 10 em.
Retention time N-acetyltryptophan = about 29 min;
Drying In an oven at 100-105 °C for 15 min.
=
1,1'-ethylidenebis(tryptophan) about 34 min.
Detection Examine in ultraviolet light at 254 run. System suitability Reference solution (c):
System suitability Reference solution (b): - resolution: minimum 8.0 between the peaks due to
- the chromatogram shows 2 clearly separated spots. N-acel)/Itryptophan and 1,1'-ethylidenebis(tryptophan);
Results The principal spot in the chromatogram obtained if necessary, adjust the time programme for the elution
with the test solution is similar in position and size to the gradient (an increase in the duration of elution with
principal spot in the chromatogram obtained with reference mobile phase A produces longer retention times and a
solution (a). better resolution);
D. Dissolve about 2 mg in 2 mL of waterR. Add 2 mL of - symmetryfactor: maximum 3.5 for the peak due to
dimethylaminobenzaldehyde solution R6. Heat on a water-bath. I,I ' -ethylidenebistryptophan in the chromatogram
A blue or greenish-blue colour develops. obtained with reference solution (c).
E. It gives the reaction of acetyl (2.3.1). Proceed as described Limits:
for substances hydrolysable only with dilficulry. - impurities A, B, C, D, E, F, G, H, I, J, K, L: for each
impurity, not more than 0.25 times the area of the
Appearance of solution reference solution (a) (0.25 per cent);
The solution is clear (2.2.1) and not more intensely coloured - total: not more than 0.5 times the area of the principal
than reference solution Y7 or GY7 (2.2.2, 1Wethod Ii). peak in the chromatogram obtained with reference
Dissolve 1.0 g in a 40 gIL solution of sodium hydroxide Rand solution (a) (0.5 per cent);
dilute to 100 mL with the same alkaline solution. - disregard limit: 0.01 times the area of the principal peak
Optical rotation (2.2.7) the chromatogram obtained with reference solution (a)
-0.1 0 to + 0.1 0 • (0.01 per cent).
Dissolve 2.50 g in a 40 gIL solution of sodium hydroxide R Ammonium (2.4.1, Method B)
and dilute to 25.0 mL with the same alkaline solution. Maximum 200 ppm, determined on 0.10 g.
Related substances Prepare the standard using 0.2 mL of ammonium standard
Liquid chromatography (2.2.29). Prepare the tes: and reference solulion (100 ppm NH.,) R.
solutions immediately before use. Iron (2.4.9)
Buffer solution pH 2.3 Dissolve 3.90 g of sodium dihydrogen Maximum 10 ppm.
phosphate R in 1000 mL of water R. Add about 100 mL of a Dissolve 1.0 g in 50 mL of hydrochlori< acidRI, with heating
2.9 gIL solution of phosphoric acid Rand adjust to pH 2.3 at 50 DC. Allow to cool. In a separating funnel, shake with
with the same acid solution. 3 quantities, each of 10 mL, of methylisobutyl ketone RI,
Solvent mixture acewnilrile R, waterR (10:90 VII'). shaking for 3 min each time. To the combined organic layers
Test solution Dissolve 0.10 g of the substance to be add 10 mL of waterR and shake for 3 min. Examine the
examined in a mixture of 50 volumes of acetonitrile R and aqueous layer.
50 volumes of waterR and dilute to 20.0 mL with the same Loss on drying (2.2.32)
mixture of solvents. Maximum 0.5 per cent, determined on 1.000 g by drying in
Reference solution (aJ Dilute 1.0 mL of the test solution to an oven at'105 °C.
100.0 mL with the solvent mixture. Sulfated ash (2.4.14)
Reference solution (b) Dilute 4.0 mL of reference solution (a) Maximum 0.1 per cent, determined on 1.0 g.
to 100.0 mL with the solvent mixture. ASSAY
Reference solution (c) Dissolve the contents ofa vial of 1,1'- Dissolve 0.200 gin 5 mL of methanol R. Add 50 mL of
ethylidenebislrypwphan CRS in I mL of reference solution (b). anhydrous ethanol R. Titrate with 0.1 1\1 sodium hydroxide,
Column: determining the end-point potentiometrically (2.2.211).
=
- size: 1= 0.25 m, 0 4.6 mID; I mL of 0.1 M sodium hydroxide is equivalent to 24.63 mg of
- stationary phase: octadecylsilyl silica gelfor chromatography R C. 3H,.,N,03'
(5 ~m);
- temperature: 40 "C. STORAGE
Mobile phase:
- mobile phase A: acetonitrile R, buffer solution pH 2.3 IMPURITIES
(115:885 VII'); Specified impuriues A, B, C, D, E, F, G, H, I, J, K, L
- mobile phase B: acetonitrile R, buffer solution pH 2.3
(350:650 VII');
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1-70 Acetyltyrosine
2022
c;:x
r '"
~
HN \ H lH,
CO,H
A. (S)-2-amino-3-(IH-indol-3-yl)propanoic acid
I. I_methyl_l,2,3,4_tetrahydro-9H_~_carboline_ 3-carboxylic
(tryptophan),
acid,
o
, HO
d¥
N H .NH
/I ,~ ' andeplmer at C*
'I : C0 2H
~ OH
B. (S)_2_amino_3_[(3RS)_3_hydroxy_2_oxo_2,3_dihydro_lH_
indol-3-yl]propanoic acid (dioxyindolylalanine),
CO,H
NHz 0 H NHz
~CO'H J. (S)-2-amino-3- [2- [2,3-dihydroxy-l-( I H-indol- 3-yl)

propylj-lH-indol-3-yl]propanoic acid,
c. (S)_2_amino_4_(2_aminophenyl)_4-oxobutanoic acid
(kynurenine),
I
¢1Y
HN H fNH,
r '" ~
CO,H CD,H
HO K. (S)_2_amino_3_[2_(IH_indo!_3_ylmethyl)_IH_indol_3_ylj
propanoic acid,
D. (S)-2-amino-3-(5-hydroxy-lH-indol-3-yl)propanoic acid
(5-hydroxytryptophan), .
OHC,
NH 0 H N~
~CO'H
E. (S)_2_amino_4_[2_(formylamino)phenylj-4-oxobutanoic
acid (N-formylkynurenine), L. 1_(IH_indol_3_ylmethyl)_1,2,3,4_tetrahydro_9H_~_
carboline-3-carboxylic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
F. (S)_2_amino_3_(phenylamino)propanoic acid Acetyltyrosine

(3-phenylaminoalanine),
(N-Acecyltyrosine, Ph. Eur. monograph 1384)
OH
I
6fY
r '" ~
N H ....NH,
Co,H
G. (S)_2_amino_3_(2_hydroxy_lH_indol_3_yl)propanoic acid
(2-hydroxytryptophan), CIlH,,NO, 223.2 537-55-3
PIlE" _
k--il1.7
~C02H
H DEFINITION
(2S)-2-(Acetylamino)-3-( 4-hydroxyphenyl)propanoic acid.
Content
H. (3RS)_1,2,3,4_tetrahydro_9H_~_carboline_3_carboxylic CHARACTERS
acid, Appearance
White or almost white, crystalline powder or colourless
crystals.
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2022 Acetyltyrosine 1-71
Solubility Mobile phase:

Freelysoluble in water, practically insoluble in cyclohexane. - mobile phaseA: mix 1.0 mL of phosphoric acid Rand
IDENTIFICATION 1000 mL of waterfor chromatography R;
- mobile phase B: acetonitrile Rl;
Second identification: A, C, D. Time Mobile phase A Moblle phase B
A. Specific optical rotation (see Tests). (min) (per cent Vm (percent Vm
B. Infrared absorption spectrophotometry (2.2.24). 0-2 97 3
2· 15 97 -t 62 3 38
Comparison N-tuetyltyrosine CRS. -t

Testsolution Dissolve 80 mg of the substance to be
examined in a mixture of 3 volumes of glacial acetic acidR, Detection Spectrophotometer at 219 nm.
3 volumes of water Rand 94 volumes of anhydrous ethanol R, Injection 2 ilL of the test solution and reference
and dilute to 10 mL with the same mixture of solvents. solutions (a), (c) and (d).
Reference solution Dissolve 80 mg of N-atetyllyrosine CRS in Relative retention With reference to N-acetyltyrosine
a mixture of 3 volumes of glacial aark acidR, 3 volumes of (retention time = about 6 min): impurity A = about 0.5.
water Rand 94 volumes of anhydrous ethanol R, and dilute to System suitability Reference solution (d):
10 mL with the same mixture of solvents. - resolution: minimum 5.0 between the principal peak and
Plate TLC silica gel F m plateR. the peak due to impurity A.
lvlobile phase water R, glacial acetic acid R, ethyl acetate R Limits:
(10:15:75 VIVIV). ---' impurity A: not more than 0.8 times the area of the
ApplicalWn 5 fIl.. corresponding peak in the chromatogram obtained with
Development Over 213 of the plate.
Drying In air. area of the principal peak in the chromatogram obtained
Detection Examine in ultraviolet light at 254 run. with reference solution (a) (0.10 per cent);
Results The principal spot in the chromatogram obtained - total: maximum 1.0 per cent;
with the test solution is similar in position and size to the - disregard limit: 0.5 times the area of the principal peakin
principal spot in the chromatogram obtained with the the chromatogram obtainedwith reference solution (a)
reference solution. (0.05 per cent).
D. Solution S (see Tests) is strongly acid (2.2.4). Chlorides (2.4.4)
Maximum 200 ppm.
TESTS
Solution S Dilute 10 mL of solution S to IS mL with water R.
Dissolve 2.50 g in waler R and dilute to 100.0 mL with the Sulfutes (2.4.13)
same solvent. Maximum 200 ppm.
Appearance of solution Dissolve 1.0 g in distilled water R and dilute to 20 mL with
Solution S is clear (2.2.1) and colourless (2.2.2, Metfwd II). the same solvent.
Specific optical rotation (2.2.7) Anunonlum (2.4.1, Metfwd B)
+ 46 to + 49 (dried substance). Maximum 200 ppm, determined on 0.100 g.
Dilute 10.0 mL of solution S to 25.0 mL with water R. Prepare the standard using 0.2 mL of ammonium standard
Related substances solution (100 ppm NH.,J R.
Liquid chromatography (2.2.29). Corry oiu the test protected Iron (2.4.9)
from light. Maximum 20 ppm.
Test solution Dissolve 50.0 mg of the substance to be In a separating funnel, dissolve 0.5 g in 10 mL of dilute
examined in mobile phase A and dilute to 50.0 mL with hydrochloric acid R. Shakewith 3 quantities, each of 10 mL,
mobile phase A. of methylisobutyl ketene Rl, shaking for 3 min each time.
Reference sduuon (a) Dilute 1.0 mL of the test solution to To the combined organic layers add 10 mL of water Rand
100.0 mL with mobile phase A. Dilute 1.0 mL of this shakefor 3 min. The aqueous layer complieswith the test.
solution to 10.0 mL with mobile phaseA. Loss on drying (2.2.32)
Reference sduuon (b) Dissolve 20.0 mg of tyrosine CRS Maximwn 0.5 per cent, determined on 1.000 g by drying in
(impurity A) in 2 mL of a 40 gIL solution of sodium an oven at 105 "C,
hydroxide R and dilute to 20.0 mL with waterR. Dilute Sulfated ash (2.4.14)
1.0 mL of this solution to 10.0 mL with water R. Maximum 0.1 per cent, determined on 1.0 g.
Reference solution (c) Dilute 1.0 mL of reference solution (b) Bacterial endotoxins (2.6.14)
to 10.0 mL with mobile phase A. Less than 25 illig, if intended for use in the manufacture of
Reference solution (d) Dilute 1.0 mL of reference parenteral preparations without a further appropriate
solution (b) to 20.0 mL with the test solution. procedure for the removal of bacterial endotoxins.
Column: ASSAY
- size: 1= 0.15 m, 0 = 3 mm; Dissolve 0.180 g in 50 mL of carbon dioxide-free waterR.
- stationary phase: spherical octade<y/si/yl silica gelfor Titrate wilh 0.1 lW sodium hydroxide, determining the
chromawgraphy R (311m); end-point potentiometrically (2.2.2(f).
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1-72 Aciclovir 2022
1 mL of 0.1 M sodium hydroxide is equivalent to 22.32 mg of CHARACTERS

C llH 13N04 · Appearance
STORAGE White or almost white, crystalline powder.
Protected from light. If the substance is sterile, store in a Solubility
sterile, airtight) tamper-evident container. Slightly soluble in water, very slightly soluble In ethanol
(96 percent), practically insoluble in heptane. It dissolves in
IMPURITIES
dilute solutions of mineral acids and alkali hydroxides.
Specified impurities A.
Otherdetectable impurities (the following substonas would, if IDENTIFICATION
present at a sufficient level, be detected by one or other of the tests Infrared absorption spectrophotometry (2.2.24).
in the monograph. They are limited by thegeneral aeuptance Comparison aciclmnr CRS.
cruetion for other/unspecified impun'ties and/or by the general TESTS
monograph Substances for pharmaceutical use (2034). It is Appearance of solution
therefore not netessary eo identify these ,mpun'ties for The solution is clear (2.2.1) and not more intensely coloured
demonstration of compliance. See also 5. JO. Control of impurities than reference solution Y7 (2.2.2J Method II).
in substances for pharmaceutical use) B.
Dissolve 0.25 g in a 4 gIL solution of sodium hydroxide Rand
H0'Qy
I "" H"N'''
dilute to 25 mL with the same solvent.
Related substances
GO,H Liquid chromatography (2.2.29). Prepare the ,oIutwns
immediauly before use.
A. (2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid Solvent mixture dimethYl sulfllXide R, water R (20:80 VIV).
(tyrosine), Phosphate buffer ,oIution pH 2.5 Dissolve 3.48 g of
dipotas,ium hydrogen phosphate R in 1000 rnL of waterfor
chromatography R and adjust to pH 2.5 with phosphoric
acid R.
Phosphate buffer solution pH 3.1 Dissolve 3.48 g of
dipotassium hydrogen pho,phate R in 1000 rnL of waterfor
chromatography R and adjust to pH 3.1 withpho,phorie
B. (2S)-2-(acetylamino)-3-[4-(acetoxy)phenyl]propanoic acid acid R.
(dlacerylryrosine). Test solution Dissolve 25 mg of the substance to be
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEur examined in 5.0 rnL of dimethYl ,ulfoxide R and dilute to
25.0 mL with water R.
Reference solution (a) Dissolve 5 mg of acidooir for system
suitobl1iry A CRS (containing impurities B, J, K, N, 0 and P)
Aciclovir in I mL of dimethYl sulfoxide R and dilute to 5 mL with
water R.
(ph. Eur. monograph 0968) Reference ,oIution (b) Dilute 1.0 mL of the test solution to
Reference solution (c) Dissolvethe contents of a vial of
acidmnrfor impurity C identification CRS in 200 Ill- of dimethyl
,ulfoxide R and dilute to 1 mL with water R.
Reference sdution (d) Dissolve the contents of a vial of
acidovirfor impun'ty G identification CRS in I mL of reference
225.2 59277-89-3
solution (a).
Action and use Co/umn:
Purine nucleoside analogue; antiviral (herpesviruses). - size: 1= 0.25 m, 0 = 4.6 mm;
Preparations - 'totionaryphase: end-capped octadeeylsi(ylsilica gelfor
Aciclovir Cream chromatographY R (5 pm).
AcicIovir Eye Ointment Mobile phase:
- mobile phase A: acewm·trile R, phosphate buffer solution
Aciclovir Infusion pH 3.1 (1:99 VIV);
Aciclovir Oral Suspension - mobile phase B: acetonitrile R, phosphate buffer solution
Adclovir Tablets pH 2.5 (50:50 VIII);
Aciclovir Dispersible Tablets
PhEv _ (min) (per cent VIV,) (per cent VII')
DEFINITION 0-5 100 o

5 - 27 100 80 0---+20
8.
--->
2-Amino-9-[(2-hydroxyethoxy)methyl]-1,9-dihydro-6H-purin-
27 - 40 2.
6-one.
Content
Flow rate 1.0 mllmin.
98.5 percent to 101.0 per cent (anhydrous substance).
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2022 Aciclovir 1-73
Injection 10 /-lL of the test solution and reference Otherdetectable impurities (thefollowing substances would, if
solutions (b), (c) and (d). present at a sufficient level, be detected by one or otherof me tests
Identification of impurities Use the chromatogram supplied in the monograph. They a~ limited by the general acceptance
with acidovirfor impun"ty C identification CRS and the criterion for other/unspecified impurities and/or by the general
chromatogram obtained with reference solution (c) to identify monograph Substances for pharmaceutical use (2034). II is
the peak due to impurity C ; use the chromatograms therefore not necessary 10 idemify these imptmues for
supplied with aciclovir for system ,uilability A CRS and demonstration of compliance" See also 5.10" Conrrol of impurities
acidovirfor impun"ly G identification CRS and the in substances for pharmaceutical use) A, F, G, 1, L, 1\-1.
chromatogram obtained with reference solution (d) to
a
identify the peaks due to impurities B, G,], K, N, 0 and P.
~N a
Rdative retention Widl reference to aciclovir (retention
HN, jl > 0---<
=
time about 13 min): impurity B = about 0.4;
impurity P = about 0.7; impurity C = about 0.9;
HzNAN N ~ CH 3
'---0
=
impurity N = about 1.37; impurities 0 and Q about 1.42;
=
impurity J about 1.62; impurities K and R = about 2.5; A. 2-[(2-amino-6-oxo-I,6-<1ihydro-9H-purin-9-yl)methoxy]
impurity G = about 2.6. . ethyl acetate,
System suitability:
- resolution: minimwn 1.5 between the peaks due co
impurity C and aciclovir in the chromatogram obtained
with reference solution (c)j minimum 1.5 between the
peaks due to impurities K and G in the chromatogram
obtained with reference solution (d)"
Limits: B. 2-amino-J,7-dihydro-6H-purin-6-one (guanine),
- C<J1'1Ution factor. for the calculation of content, multiply the
peak area of impurity C by 2.2j
- impurity B: not more than 7 times the area of the
- impun"ty J: not more than twice the area of the principal
peak in the chromatogram obtained with reference C. 2-amino-7-[(2-hydroxyethoxy)methyl]-I,7-dihydro-6H-
solution (b) (0.2 per cent); purin-e-one,
- sum of impurities K and R: not more than 1.5 times the
with reference solution (b) (0.15 per cent);
- sum of impurities 0 and Q: not more man 1.5 times the
- impurities C, N, P: for each impurity, not more than
F. N-[9-[(2-hydroxyethoxy)methyl]-6-oxo-6,9-<1ihydro-IH-
purin-2-yl]acetamide,
chromatogram obtained with reference solution (b)
(0.15 per cent); o
- unspecified impurities: for each impurity, not more than ~N
HN , j l >
0
0.5 times the area of the principal peak in the a 0---<
chromatogram obtained with reference solution (b) H,C)lN~N N ~ CH,
(0.05 per cent); H '---0
- total: not more than 10 times the area of the principal
peak in the chromatogram obtained with reference G.2-[(2-acetamido-6-oxo-I,6-dihydro-9H-purin-9-yl)
solution (b) (1.0 per cent); methoxy]ethyl acetate,
(0.03 per cent).
Water (2.5.12)
ASSAY I. 2-amino-7-[[2-[(2-amino-6-oxo-I,6-dihydro-9H-purin-9-
yl)methoxyJ ethoxy1methyl] -1,7-dihydro-6H-purin-6-one,
Dissolve 0.150 g in 60 mL of anhydrous acetic acidR. Titrate
with 0"1 .M perchloric add, determining me end-point
potentiometrically (2.2.20). Carry out a blank tittation.
I mL of 0.1 M perchloric acid is equivalent to 22.52 mg
of CBHlINsOj.
L"liPURITIES
Specified impuriues B, C, J, K,. N, 0, P, Q, R. J. 9,9'-[ethane-I,2-diylbis(oxymelhylene)]bis(2-amino-l ,9-
dihydro-6H-purin-6-one),
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1-74 Acitrerin 2022
Acitretin
(Ph. Eur. monograph 1385)
K. 2,2'-(methylenediazanediyl)bis[9-[(2-hydroxyethoxy) CD,H
methyl]-I ,9-dihydro-6H-purin-6-one],
",CO
CH,
326.4 55079-83-9
Action and use

Vitamin A analogue (retinoid); treatment of psoriasis;
ichthyosis; Darier's disease.
L. N-(9-acetyl-6-oxo-6,9-dihydro-IH-purin-2-yl)acetamide
(N',9-diacetylguanine). Preparation
Acitretin Capsules
o
o 0 0 /I PhEIr _
~/'--J\
Jl 1,l N> CH,
DEFINITION
(2E,4E,6E,8E)-9-( 4-Methoxy-2,3,6-trimethylphenyl)-3,7-
~c N N N
H dimethylnona-2,4,6,8-tetraenoic acid
Content
,,,1. 2-[(2-acetamido-6-oxo-I,6-dihydro-7H-purin-7-yl) 98.0 per cent [Q 102.0 per cent (dried substance).
methoxy]ethyl acetate,
CHARACTERS
N. unknown structure,
Appearance
O. unknown structure, Yellow or greenish-yellow, crystalline powder.
Solubility
Practically insoluble in water, sparingly soluble in
tetrahydrofuran, slightly soluble in acetone and in ethanol
(96 per cent), very slightly soluble in cyclohexane.
It is sensitive to air, heat and light, especially in solution.
P. 2-amino-9-(2-hydroxyethyl)-1,9-dihydro-6H-purin-6-one,
Carry out all operations as rapidly as possible and avoid exposure
to actinic lighlj use freshly prepared solutions.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
Comparison acuretin CRS.
substance separately in 2-propanol R by heating under reflux;
filter, evaporate to dryness and recordnew spectra using the
residues.
TESTS
Related substances
Q. mixture of 2-amino-9-[[2-(hydroxyethoxy) Liquid chromatography (2.2.29). Cany out the ,,$I protected
methoxy]methyl]-1,9-dihydro-6H-purin-6-one and from light andprepare the solutions immediately before use.
2-amino-9-[[2-(hydroxymethoxy)ethoxy]methyl]-1,9-
Test solution (a) Dissolve 25.0 mg of the substance to be
dihydro-6H-purin-6-one,
examined in 5 mL of utrahydrofuran R and dilute [0
100.0 mL with anhydrous ethanol R.
Test sduiion (b) Dilute 10.0 mL of test solution (a) to
25.0 mL with anhydrous ethanol R.
Reference solution (a) Dissolve 25.0 mg of acureti:.. CRS in
5 mL of seuanydrofunm R and dilute to 100.0 mL with
a..hydrous ethanolR. Dilute 10.0 mL of the solution to
25.0 mL with anhydrous ethanolR.
R. 9,9'-[methylenebis(oxyethane-2,I-diyloxymethylene)]bis Reference solution (b) Dissolve I mg of ,retinoin CRS in
(2-amino-I,9-dihydro-6H-purin-6-one). anhydrous ethanol R and dilute to 20.0 mL with the same
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" solvent. Mix 5.0 mL of the solution with 2.5 mL of reference
solution (a) and dilute to 100.0 mL with anhydrous ethanol R.
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2022 Adapalene 1-75
Reference solution (e) Dilute 1.0 mL of the test solution (a) in the monograph. They are limited try thegeneral acceptance
to 100.0 mL with anhydrous ethanol R. Dilute 1.0 mL of this cruerion for other/unspecified impurities and/or by thegeneral
solution to 10.0 mL with anhydrous ethanol R. monograph Substances for pharmaceutical use (2034). It is
Reference sdution (d) Dissolve 2.5 mg of acitretin for therefore not necessary UJ identiJY these impurities for
impurity A identification CRS in 0.5 mL of tetrahydrofuran R demonstration of compliaru:e. See also 5,10, Control of impurities
and dilute to 10.0 mL with anhydrous ethanol R. in substances for phannaceutica/ use) B,
Column:
- size: 1= 0.25 m, III = 4 mm;
- stationary phase: octadecylsilyl silica gel for
chromatography for separation of polycyclic aromatic
hydrocarbons R (5 urn);
- temperature: 25°C.
Mobilephase 0.3 per cent VIV solution of glacial acetic
acid R in a mixture of 8 volumes of waterfor A. (2Z,4E,6E,8E)-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-
chromatography Rand 92 volwnes of anhydrous ethanol R. dimethylnona-2,4J6,8-tetraenoic acid,
Flow rate 0.6 mlJrnin. CH,
Auunampler Set at 4°C.
Injection 10 J.tL of test solution (a) and reference H,CO CH,
solutions (b), (c) and (d). CH,
Run time 2.5 times the retention time of acitretin.
Identification of impun"t;es Use the chromatogram supplied B. ethyl (2E,4E,6E,8E)-9-(4-methoxy-2,3,6-trimethylphenyl)-
with acitretin for ;mpun'ty A identification CRS and the 3,7 -dimethylnona-2,4,6,8-tetraenoate.
chromatogram obtained with reference solution (d) to _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
identify the peak due to impurity A.
Relativeretention With reference to acitretin (retention
=
time about 6 min): impurity A about 0.8;=
tretinoin ;;;; about 0.85. Adapalene
System suitability Reference solution (b):
- resolution: minimum 2,0 between the peaks due to (ph. Eur. monograph 2445)
tretinoin and acitretin.
- for each impurity, use the concentration of acitretin in
reference solution (c).
Limits:
- impun'ty A: maximwn 0.2 per cent;
- unspecified impun'ties: for each impurity, maximum
0.10 per cent;
- total: maximum 0.5 per cent; 412.5 106685-40-9
- reporting threshold: 0.05 per cent.
Action and use
Lo ss on drying (2.2.32)
Vitamin A analogue (retinoid); treatment of acne.
vcu:uo at 100°C for 4 h. Preparadons
Adapalene Cream
Maximum 0,1 per cent, determined on 1.0 g. Adapalene Gel
ASSAY PhE" _
Liquid chromatography (2.2.29) as described in the test for DEFINITION

related substances with the following modification. 6-(4-Methoxy-3-tricyc!0[3.3.I.l"'Jdec-l-ylphenyl)
Injection Test solution (b) and reference solution (a). naphthalene-2-earboxylic acid.
Calculate the percentage content of C21H260j taking into Content
account the assigned content of acureun CRS. 98.0 per cent to 102.0 per cent (dried substance).
STORAGE CHARACTERS
In an airtight container, protected from light, at a Appearance
temperature of 2 °C to 8°C, White or almost white powder.
It is recommended that the contents of an opened container Solubility
be used as soon as possible and any unused part be protected Practically insoluble in water, sparingly soluble in
by an atmosphere of lnerr gas, tetrahydrofuran, practically insoluble in ethanol
IMPURITIES (96 per cent).
Specified impurilies A. IDENTIFICATION
Otherdetectable impurities (thefollowing sub'tanus would, if Infrared absorption spectrophotometry (2.2.24).
present at a sufficient level, be detected by one or other of the tests Comparison adapaime CRS.
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1-76 Adapalene 2022
TESTS Relativeretention With reference to adapalene (retention

Appearance of solution ume e about 20 min): impurity A::: about 0.3j
The solution is clear (2.2.1) and not more intensely coloured impurity C =about 0.9; impurity D =about 1.9.
than reference solution BY6 (2.2.2, lHethod II). System suiwbility Reference solution (b):
Dissolve 0.2 g in tetrahydrofuran R and dilute to 20 mL with - resolution: minimum 4.5 between the peaks due to
the same solvent. impurity C and adapalene;
Related substances - signal-to-noise ratio: minimum 10 for the peak due to
Liquid chromatography (2.2.29). impurity C.
Solvent mixture tetrahydrofuran R, acetonitrile R, water R Limits:
(20:37:43 VIVIII). - correction factors: for the calculation of content, multiply
the peak areas of the following impurities by the
corresponding correction factor: impurity A = 0.7;
examined in 10 mL of tetrahydrofuran R, add 7 mL of the
solvent mixture and dilute to 20.0 mL with tetrahydrofuran R.
impurity C =7; impurity D =1.4;
- impun'ty A: not more than 3 times the area of the
Test solution (b) Dissolve 20.0 mg of the substance to be principal peak in the chromatogram obtained with
examined in 50 mL of tetrahydrofuran R, add 35 mL of the reference solution (a) (0.3 percent);
solvent mixture and dilute to 100.0 mL with - impurity D: not more than twice the area of the principal
tetrahydrofuran R. Dilute 5.0 mL of the solution to 50.0 mL peakin the chromatogram obtained with reference
with the solvent mixture. solution (a) (0.2 per cent);
Reference solution (a) Dilute 1.0 mL of test solution (a) to - impun'cy C: not more than 1.5 times the area of the
10.0 mL with tetrahydrofuran R. Dilute 1.0 mL of this principal peak in the chromatogram obtained with
solution to 100.0 rnL with the solvent mixture. reference solution (a) (0.15 per cent);
Reference solution (b) Dissolve 2.4 mg of adapalene - unspedfied impun·tieJ: for each impurity, not more than the
impurity C CRS in 2 mL of tetrahydrofuran R and dilute to area of the principal peak in the chromatogram obtained
20.0 mL with the same solvent. Dilute 2.0 mL of the with reference solution (a) (0.10 percent);
solution to 20.0 mL with the solvent mixture. To 2.0 mL of - total: not more than 5 times the area of the principal peak
this solution add 2.0 mL of reference solution (a) and dilute In the chromatogram obtained with reference solution (a)
to 20.0 mL with the solvent mixture. (0.5 per cent);
Reference solution (c) Dissolve the contents of a vial of - disregard limit: 0.5 rimes the area of the principal peak in
adapaknefor peak idemlfication CRS (containing impurities A, the chromatogram obtained withreference solution (a)
(0.05 per cent).
C and D) in 0.5 mL of teti'ahydrofuran R and dilute to
1.0 mL with the solvent mixture. Loss on drying (2.2.32)
Reference solution (d) Dissolve 20.0 mg of adapolene CRS in Maximum 0.5 per cent, determined on 1.000 g by drying in
50 mL of tetrahydrofuran R, add 35 mL of the solvent an oven at 105 °C for 4 h.
mixture and dilute to 100.0 mL with tetrahydrofuran R. Sulfated ash (2.4.14)
Dilute 5.0 mL of the solution to 50.0 mL with the solvent Maximum 0.1 percent, determined on 1.0 g.
mixture. ASSAY
Column: Liquid chromatography (2.2.29) as descnbed in the test for
- size: I::: 0.25 m, 0 ::: 4.6 mID; related substances with the following modification.
- stationary phase: end-eapped phenyfsilyl SIlica gelfor
Iniecticn Test solution (b) and reference solution (d).
chromatography R (5 pm) with a carbon loading of
7.5 percent; Calculate the percentage contentof adapalene from the
- temperature: 30°C. declared contentof adopolene CRS.
Mobile phase: IMPURITIES
- mobile phaseA: glacial acetic add R, waterR (0.1: 100 VIII); Specified impurities A, C, D.
- mobile phase B: tetrahydrofuran R, acetonitrile R Otherdete<table impurities (thefollowing substances would, if
(35:65 VIII); present at a suffidentlevel, be detected by one or other of the tests
Time MobUe phase A Mobile phase B criterion for otherlunspedfied impuritks and/or by thegeneral
(min) (per cent VIJI) (per cent VIJi)
monograph Substances for pharmaceutical use (2034). It is
0-2.5 50 50 therefore not necessary to idenuJy these impmities for
2.5 - 40 50 ---> 28 50 ---> 72 demonstration of compliance. See also 5.10. Ccmtrol of impurides
40 - 42 28 72 in substances for pharmaceutical use) B.
Flow rate 1.2 mUmin. HD,C

Delation Spectrophotometer at 270 om.
Injection 25 j.tL of test solution (a) and reference
solutions (a), (b) and (c).
Identification of impurities Use the chromatogram supplied
CD,H
with adapalene for peak identijicaticn CRS and the
chromatogram obtained with reference solution (c) to identify
A. 2,2'-binaphthalene-6,6/-dicarboxytic acid,
the peaks due to impurities A, C and D.
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2022 Adenine 1-77
add 15 mL of lightpetroleum R and heat to boiling with

vigorous stirring. Cool and filter, Wash the precipitate with
two quantities, each of 5 mL, of lightpetroleum R. Dissolve
the precipitate in 10 mL of wacer R and boil for 1 min. Filter
the mixrure at 30 °C to 40 °C. Allow to cool. Filter, and dry
OH
Co,H me precipitate at 100 °C to 105 °C for 1 h. The melting
point (2.2.14) of the precipitate is 231 "C to 241 "C.
B. 6-[3-(3-hydroxyuicyclo[3.3. I. I>.7jdec-I-yl)-4-
TESTS
methoxyphenyl]naphthalene-2-earboxylic acid,
Solution S
Suspend 2.5 g in 50 mL of distiUed water R and boil for
3 min. Cool and dilute to 50 mL with distil/ed water R. Filter.
Use the filtrate as solution S.
Dissolve 0.5 g in dilute hydrochloric acid R and dilute to
C. 1-(2-methoxyphenyl)cricyclo[3.3.1.I"'jdecane, 50 mL with the same acid. The solution is clear (2.2.1) and
colourless (2.2.2, J.Hethod II).
Acidity or alkalinlty
To 10 mL of solution S add 0.1 mL of bromothymol blue
solution Rl and 0.2 mL of 0.01 M sodium hydroxide.
OCH, The solution is blue. Add 0.4 mL of 0.01 M hydrochloric acid.
The solution is yellow,
D. 1,1'-[4,4'-bis(methoxy)biphenyl-3,3'-diyljbis(tricyclo Related substances
[3.3.1.1'·7jdecane). Examine by thin-layer chromatography (2.2.27), using silica
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw
gel GFZ54 R as the coating substance.
Test solution (aJ Dissolve 0.10 g of the substance to be
examined in dilute acetic add R, with heating if necessary, and
dilute to 10 mL with the same acid.
Adenine Test solution (b) Dilute I mL of test solution (a) to 10 mL
with dilute acetic acid R.
(ph. Bur. monograph oso» Reference solution (a) Dissolve 10 mg of adenine CRS In
NH, dilute acetic add R, with heating if necessary, and dilute to
N~~ 10 mL with the same acid.
1lJ-N>
Reference solution (b) Dilute 1 mL of test solution (b) to
N 20 mL with dilute acetic acid R.
Reference sdution (e) Dissolve 10 mg of adenine CRS and
C,H,N, 135.1 73-24-5 10 mg of adenosine R in dilute acetic acid RJ with heating if
necessary, and dilute (0 10 mL with the same acid.
Action and use Apply to the plate 5 J.lL of each solution. Develop over a
Constituent of anticoagulant and preservative solutions for
path of 12 cm using a mixture of 20 volumes of concentrated
blood.
ammonia R J 40 volumes of ethyl euetate R and 40 volumes of
PhE" ~ propanol R. Dry the plate in a current of warm air and
examine in ultraviolet light at 254 run. Any spot in the
DEFINITION chromatogram obtained with test solution (a), apart from the
Adenine contains not less than 98.5 per cent and not more principal spot, is not more intense than the spot in the
than the equivalent of 101.0 per cent of7H-purin-6-amine, chromatogram obtained with reference solution (b)
calculated with reference to the dried substance. (0.5 per cent). The test is not valid unless the chromatogram
CHARACTERS obtained with reference solution (c) shows two dearly
A white or almost white powder, very slightly soluble in separated spots.
water and in alcohol. It dissolves in dilute mineral acids and Chlorides (2.4.4)
in dilute solutions of alkali hydroxides. To 10 mL of solution S add 1 mL of concentrated ammonia R
IDENTIFICATION and 3 mL of silver nitrate solution R2. Filter. Wash the
precipitate with a little water Rand dilute the filtrate to
First identification: A.
J 5 mL with water R. The solution complies with the limit
Second identification: B, C. test for chlorides (100 ppm). When carrying out the test, add
A. Examine by infrared absorption spectrophotometry 2 mL of dilute nit,;c add R instead of 1 mL of dilute nioic
(2.2.24), comparing with the spectrum obtained with acid R.
adenine CRS. Examine the substances prepared as discs.
Sulfates (2.4.13)
B. Examine the chromatograms obtained in the test for Dilute 10 mL of solution S to 15 mL with distilkd waterR.
related substances. The principal spot in the chromatogram The solution complies with the limit test for sulfates
obtained with test solution (b) is similar in position and size (300 ppm).
to the principal spot in the chromatogram obtained with
reference solution (a). Ammonium
Prepare a cell consisting of two watch-glasses 60 mm in
C. To I g add 3.5 mL of propionic anhydrUk R and boil for
diameter placed edge to edge. To the inner wall of the upper
15 min with stirring. Cool. To the resulting crystalline mass
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1-78 Adenosine 2022
watch-glass stick a piece of red litmus paper R 5 mm square TESTS

and wetted with a few drops of water R. Finely powder the Solution S
substance to be examined, place 0.5 g in the lower watch- Suspend 5.0 g in 100 mL of distilled water R and heat to
glass and suspend in 0.5 mL of water R. To the suspension boiling. Allow to cool, filter with the aid of vacuum and
add 0.30 g of heavymagnesium oxide R. Briefly triturate with dilute to 100 mL with distilled waterR.
a glass rod. Immediately close the cell by putting the two Appearance of solution
watch-glasses together. Heat at 40 °C for 15 min. The litmus Solution S is colourless (2.2.2, Method If).
paper is not more intensely blue coloured man a standard
prepared at the same time and in me same manner using Acidity or alkalinlty
To 10 mL of solution S, add 0.1 mL of bromocresol purple
0.05 mL of ammonium standard solution (100 ppm NH,) R,
0.5 mL of water Rand 0.30 g of heavy magnesium oxide R
solution Rand 0.1 mL of 0.01 M hydrochloric acid.
The solution is yellow. Add 0.4 mL of 0.01 M sodium
(10 ppm).
hydroxide. The solution is violet-blue.
Not more than 0.5 per cent, determined on 1.000 g by
-45 to -49 (dried substance).
drying in an oven at 105 "C.
Dissolve 1.25 g in 1 M hydrochloric acid and dilute to
50.0 mL with the same acid. Examine within 10 min of
Not more then 0.1 per cent, determined on 1.0 g.
preparing the solution.
ASSAY Related substances
Dissolve 0.100 g in a mixture of 20 mL of acetic anhydride R Liquid chromatography (2.2.29).
and 30 mL of anhydrous acetic acidR. Titrate with 0.1 M
Solvent mixture Dissolve 6.8 g of potassium hydrogen sulfate R
percblonc acid, determining the end-point potentiometrically
and 3.4 g of teuabutylammomum hydrogen sulfate R in water R,
(2.2.2(/).
adjust to pH 6.5 with a 60 gIL solution of potassium
1 mL of 0.1 M. perchloric acid is equivalent to 13.51 mg of hydroxide R and dilute to 1000 mL with the same solvent.
C,H,Nj • Use a freshly prepared solvent mixture.
___ ~ PhE"
examined in the mobile phase and dilute to 20 mL with the
mobile phase.
Adenosine 100.0 mL with the mobile phase. Dilute 1.0 mL of this
solution to 10.0 mL with the mobile phase.
Reference solution (b) Dissolve 5 mg of adenine R
Nt<, (impurity A) and 5 mg of inosine R (impurity G) in the
NJyN) mobile phase and dilute (0 50 mL with the mobile phase.

Dilute 4 mL of this solution to 100 mL with the mobile
HO ~NJlN phase.
~
Column:
- size: 1 = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-copped octadecylsi!YI silica gellor
OH OH chromatography R (5 um).
Mobile phase waterR, solvent mixture (40:60 VIV).
267.2 58-61-7 Flow rat< 1.5 mUmin.
Action and use Detection Spectrophotometer at 254 nm.
Antiarrhythmic. Injection 20~.
PhE" _
Run time 1.5 times the retention time of adenosine.
Relativeretention With reference to adenosine (retention
DEFINITION = =
rime about 13 min): impurity A about 0.3;
9-Jl-D-Ribofuranosyl-9H-purin-6-amine. impurity G = about 0.4.
Content System suitability Reference solution (b):
99.0 per cent to 101.0 per cent (driedsubstance). - resolution: minimum 1.5 between the peaks due (0
impurities A and G.
CHARACTERS
Appearance Limits:
White or almost white, crystalline powder. - CQ17UUon facum: for the calculation of content, multiply
Solubility corresponding correction factor: impurity A = 0.6;
Slightly soluble in water, soluble in hot water, practically =
impurity G 1.4;
insoluble in ethanol (96 per cent) and in methylene chloride. - impurity A: not more than twice the area of the principal
It dissolves in dilutemineral acids. peak in the chromatogram obtained with reference
mp solution (a) (0.2 per cent);
About 234 ·C. - impun"ty G: not more than the area of the principal peak
IDENTIFICATION in the chromatogram obtained withreference solution (a)
Infrared absorption spectrophotometry (2.2.24). (0.1 per cent);
Comparison adenosine CRS.
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2022 Adipic Acid 1-79
- unspecified impurities: for each impurity, not more than the o

HN~N~
"O~
(0.5 per cent);
OH OH
(0.05 per cent).
Chlorides (2.4. f) G. 9-~-D-ribofuranosyl-l,9-dihydro-6H-purin-6-one (inosine),
Maximum 100 ppm.
o
Dilute 10 mL of solution S to 15 mL with waterR.
Sulfates (2.4.13) HN:XN~
Maximum 200 ppm, determined on solution S.
Ammonium (2.4.1, Merhod B)
H'N~AN
HO N
Maximum 10 ppm, determined on 0.5 g. o

Prepare the standard using 5 mL of ammonium standard
solution (l ppm NH.J R. OH OH
Maximum 0.5 per cent, determined on 1.000 g by drying in H.2-amino-9-ll-o-ribofuranosyl-1,9-dihydro-6H-purin-6-one
an oven at 105 "C. (guanosine).
Sulfated ash (2.4. If) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>E<I
ASSAY
Dissolve 0.200 g) warming slightly if necessary, in a mixture
of 20 mL of acetic anhydride Rand 30 mL of anhydrous acetic Adipic Acid
acidR. Titrate with 0.1 M perchloric acid, determining the
(ph. Ellr. monograph 1586)
1 mL of 0.11\1 perchknic acid is equivalent to 26.72 mg
of CIOHI:3Nj04'
IMPURITIES 146.1 124-04-9
Specified impurities A, G.
Otherdetectable impurities (the following SlIbstances mould, if Action and use
present at a sufficient leud, be detected by oneor otherof the tests Excipient.
in the monograph. They are limited by thegeneral a«eptance Pl>E<I _
criterion for other/unspecified impun"ties andlor by the general
monograph Substances for pharmaceutical use (2034). lr is DEFINITION
therefore not necessary to identify these impuritles for Hexanedioic acid.
demonstration of compliance. See also 5.10. Control of impurities Content
in substances for phamzaceutical use) FJ H. 99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
~N Appearance
tJ-N
~ ~
Solubility
Sparingly soluble in water, soluble in boiling water, freely
soluble in ethanol (96 per cent) and in methanol, soluble in
A. 7H-purin-6-amine (adenine),
acetone.
IDENTIFICATION
A. Melting point (2.2. If): 151 "C to 154 'C.
B. Infrared absorption spectrophotometry (2.2.2f).
Comparison adipic acidCRS.
TESTS
Solution S
Dissolve 5.0 g with heating in distilled waterR and dilute to
50 mL with the same solvent. Allow to cool and to
F. 1-~-o-ribofuranosylpyrimidine-2,4(IH,3H)-dione crystallise. Filter through a sintered-glass filter (40) (2.1.2).
(uridine), Wash the filter with dis.1Ied water R. Collect the filtrate and
the washings until a volume of 50 mL is obtained.
MethodII).
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1-80 Adrenaline 2022
Dissolve 1.0 g in me/hanoi R and dilute to 20 mL with the Iron (2.4.9)

same solvent. Maximum 10 ppm, determined on solution S.
Related substances Loss on drying (2.2.32)
Liquid chromatography (2.2.29). Maximum 0.2 per cent, determined on 1.000 g by drying in
Test solution Dissolve 0.20 g of the substance to be an oven at 105°C.
examined in the mobile phase and dilute to 10.0 mL with Sulfated ash (2.4.14)
the mobile phase. Maximum 0.1 per cent.
Reference solution (a) Dissolve 20 mg of glutarit: acid R in Melt 1.0 g completely over a gas burner, then ignite the
1.0 mL of the test solution and dilute to 10.0 mL with the melted substance with the burner. After ignition, lower or
mobile phase. remove the flame in order to prevent the substance from
Reference solution (b) Dilute 1.0 mL of the test solution to boiling and keep it burning until completely carbonised.
100.0 mL with the mobile phase, dilute 1.0 mL of the Carry out the test for sulfated ash using the residue.
solution to 10.0 mL with the mobile phase. ASSAY
Column: Dissolve 60.0 mg in 50 mL of water R. Add 0.2 mL of
=
- size: / 0.125 m, 0 = 4.0 mm, phenolphthalein solution Rand titrate with 0.11\-1 sodium
- stationary phase: spherical octadecylsily[ silica gelfor hydroxide.
chromatography R (5 urn) with a specific surface area of 1 mL of 0.1 M sodium hydroxide is equivalent to 7.31 mg of
350 m2/g and a pore size of 10 om, C 6H IO04 •
IMPURITIES
lvlob,7e phase Mix 3 volwnes of acetonitrile Rand
97 volumes of a 24.5 gIL solution of dihue phosphoric acid R.
Flow raUt I mUmin.
Detection Spectrophotometer at 209 run. A. pentanedioic acid (glutaric acid),
Injection 20 ~L
Run time 3 times the retention time of adipic acid.
System sU1iabllity Reference solution (a):
- resolution: minimum 9.0 between the peaks due to glutaric B. buranedioic acid (succinic acid),
add and adipic acid.
Ho,C~co,H
Limits:
- any impurity: not more than the area of the principal peak
in the chromatogram obtained with reference solution (b) C. heptanedioic acid (pimelic acid).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE"
(0.1 per cent),
- total: not more than 5 times me area of the principal peak
in the chromatogram obtained with reference solution (b)
(0.5 per cent),
- disregard limit: 0.5 times the area of the principal peak in Adrenaline I Epinephrine
(0.05 per cent). . (Ph. Eur. monograph 2303)
Chlorides (2.4.4)
H, OH H
Maximum 200 ppm.
HO~·
I N,
Dilute 2.5 mL of solution S to IS mL with water R. CH,
Nitrates HO "'"
Maximum 30 ppm.
To I mL of solution S add 2 mL of concentrated ammonia R, c.H,,NO, 183.2 51-43-4
0.5 mL of a 10 gIL solution of manganese sulfate R, 1 mL of
a 10 gIL solution of sulfamlamide R and dilute to 20 mL with Action and use
water R. Add 0.10 g of zinc powder R and cool in iced water Adrenoceptor agonist.
for 30 min; shake from time to time. Filter and cool 10 mL Preparations
of the filtrate in iced water. Add 2.5 mL of hydrochloric Adrenaline Eye DropslEpinephrine Eye Drops
acid R1 and I mL of a 10 gIL solution of Dilute Adrenaline Injection (I in 1O,000)lDilute Epinephrine
naph'hy/ethy/enediamine dihydrochlon"de R. Allow to stand at Injection (I in 10,000)
room temperature. After 15 min the mixture is not more
PIlE" _
intensely coloured than a standard prepared at the same time
and in the same manner, using 1.5 mL of nitrate standard DEFINITION
solution (2 ppm NO) R instead of I mL of solution S.
4- [(IR)-I-H ydroxy-2-(methylamino)ethyl]benzene-I,2-diol.
The test is invalid if a blank solution prepared at the same
time and in the same manner, using I mL of water R instead Synthetic product.
of 1 mL of solution S, is more intensely coloured than a Content
2 mgIL solution of potassium permanganate R. 99.0 per cent to 101.0 per cent (dried substance).
Sulfutes (2.4.13) CHARACTERS
Maximum 500 ppm. Appearance
Dilute 3 mL of solution S to 15 mL with distiUed water R. White or almost white crystalline powder, becoming coloured
on exposure to air and light.
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2022 Adrenaline 1-81
Solubility Time Mobile phase A Mobile phase B

Practically insoluble in water, in ethanol (96 per cent) and in (miD) (per cent VIJ') (per ceot VIJ')
methylene chloride. It dissolves in hydrochloric acid. 0-15 92 --> 50 8 --> 50

15 - 20 50 --> 92 50 --> 8
IDENTIFICATION 20 - 25 92 8
A. Infrared absorption spectrophotometry (2.2.24).
Comparison adrenaline CRS. Flow rate 2.0 mllmin.
B. Specific optical rotation (see Tests).
Detection Spectrophotometer at 210 run"
TESTS Injection 20 ~L.
Solution S Identification of impuruies Use the chromatogram obtained
Dissolve 1.000 g in a 25.75 gIL solution of hydrochloric acid R with reference solution (b) {Q identify the peaks due to
and dilute to 50.0 mL with the same solvent. Examine the impurities Band C; use the chromatogram supplied with
solution immediately. adrenaline impun"ty mixture CRS and the chromatogram
Appearance of solution obtained with reference solution (c) (Q identify the peaks due
Solution S is not more opalescent than reference to impurities D and Ej use the chromatogram obtained with
suspension II (2.2.1) and not more intensely coloured than reference solution (d) to identify the peak due to impurity F.
reference solution BY j (2.2.2, JUelhod 1/). Relativeretention With reference to adrenaline (retention
Specific optical rotation (2.2.7) time = about 4 min): impurity F ;;;; about 0.2j
-50.0 to -54.0 (dried substance), determined on solution S. impurity B =about 0.8j impurity C ;;;; about 1.3j
Related substances impurity D =about 3.3j impurity E = about 3.7.
Liquid chromatography (2.2.29). Prepare the ,oIutions protected System suitability Reference solution (b):
from light; - resolution: minimum 3.0 between the peaks due to
Solvent mixtureA Dissolve 5.0 g of potassium dihydrogen impurity B and adrenaline.
phosphate Rand 2.6 g of sodium octanesuljonate R in waterfor Limits:
chromatography R and dilute (0 1000 mL with the same - correction factors: for the calculation of content, multiply
solvent (it is usually necessary to stir for at least 30 min to the peak areas of the following impurities by the
achieve complete dissolution). Adjust [Q pH 2.8 with corresponding correction factor: impurity D ;;;; 0.7j
phosphoric acid R. impurity E 0.6; =
Solvent mixture B acetonitrile R, solvent mixture A - impurities B, C, F: for each impurity, not more than twice
(13:87 VIII). the area of the principal peak in the chromatogram
obtained with reference solution (a) (0.2 per cent);
Solvent mixture C 10.3 gIL solution of hydrochloric acid R, - impurities D, E: for each impurity, not more than the area
solvent ntiature B (10:90 VIII).
of the principal peak in the chromatogram obtained with
Testsolution Dissolve 40.0 mg of the substance to be reference solution (a) (0.1 per cent);
examined in 5 mL of a 10.3 gIL solution of hydrochloric - unspecified impurities: for each impurity, not more than the
acidR and dilute to 50.0 mL with solvent mixture B. area of the principal peak in the chromatogram obtained
Reference solution (a) Dilute 1"0 mL of the test solution to with reference solution (a) (0.10 per cent);
100.0 mL with solvent mixture B. Dilute 1.0 mL of this - rota/: not more than 5 times the area of the principal peak
solution to 10.0 mL with solvent mixture B. in the chromatogram obtained with reference solution (a)
Reference ,oIution (b) Dissolve 1.5 mg of noradrenaline (0.5 per cent);
lanrate CRS (impurity B) and 1.5 mg of adrenalone - disregard limit: 0.5 times the area of the principal peak in
hydrochloride R (impurity C) in solvent mixture B, add I mL the chromatogram obtained with reference solution (a)
of the test solution and dilute to 100 mL with solvent (0.05 per cent).
mixture B. Loss on drying (2.2.32)
Reference solution (c) Dissolve the contents of a vial of Maximum 0.5 per cent, determined on 1.000 g by drying in
adrenaline impun·ty mixture CRS (containing impurities D a desiccator at a pressure not exceeding 0.7 kPa for 18 h.
and E) in I mL of solvent mixture C. Sulfated ash (2.4.14)
Reference solution (d) Dissolve the contents of a vial of Maximum 0.1 per cent, determined on 1.0 g.
adrenaline impuniy F CRS in I mL of a 10.3 gIL solution of ASSAY
hydrochloric acid R. Dissolve 0.150 g in 50 mL of anhydrous acetic acid R. Titrate
Column: with 0.1 M perchloric acid, determining the end-point
- size: I;;;; 0.10 m, 0 ;;;; 4.6 mm; potentiometrically (2.2.20).
- stationary phase: base-deactivated end-copped ocraduylsi/yl
1 mL of 0.1 M perchloric acid is equivalent to 18.32 mg
silica gd for chromatography R (3 ~m);
of C 9H13N 0 3 •
- temperature: 50 °C.
Mobile phase: STORAGE
- mobile phase A: acetonitrile Rl, solvent mixture A Under nitrogen, protected from light.
(5:95 VIII); IMPURITIES
- mobile phaseB: acetonitrile Rl, solvent mixture A Specified impuruies B, G, D, E, F"
(45:55 VIII);
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1-82 Adrenaline Acid Tartrate 2022
H OH Ph,,, _
HO~NH' DEFINITION
HO.v (IR)-I-(3,4-Dihydroxyphenyl)-2-(methylamino)ethanol
hydrogen (2R,3R)-2,3-dihydroxybutanedioate.
B. (1R)-2-amino-I-(3,4-dihydroxyphenyl)ethanol
Content
(noradrenaline),
o
CHARACTERS
Hoif~'
""" CH3 Appearance
White or greyish-white) crystalline powder.
HO -'"
Solublllty
C. 1-(3,4-dihydroxyphenyl)-2-(methylamino)ethanone Freely soluble in water, slightly soluble in ethanol
(adrenalone), (96 per cent).
IDENTIFICATION
A. Dissolve 5 g in 50 mL of a 5 gIL solution of sodium
metabisulfite R and make alkaline by addition of ammonia R.
Keep the mixture at room temperature for at least 15 min
and filter. Reserve the filtrate for identification test C. Wash
the precipitate with 3 quantities, each of 10 rnl., of
D.4-[(IR)-2-(benzylmethylamino)-I-hydroxyethyl]benzene- methanol R. Dry at 80°C. The specific optical rotation
1,2-dioIJ (2.Z.7) of the residue (adrenaline base) is -53.5 to -50,
determined using a 20.0 gIL solution in 0.5 M hydrodllori<
acid.
B. Infrared absorption spectrophotometry (Z.2.24).
Preparation Discs of adrenaline base prepared as described
underidentification [est A.
Comparison Use adrenaline base prepared as descnbed
underidentification test A from 50 mg of adrenaline
E. 2-(benzylmethylamino)-I-(3,4-dihydroxyphenyl)ethanone, tartrate CRS dissolved in 5 mL of a 5 gIL solutionof sodium
metabisulfite R. Keep the mixture at room temperature for at
HO~~,CH' least 30 min. Filter through a sintered-glass filter (2.1.2).

C. 0.2 mL of the filtrate obtained in identification test A
HO.v
gives reaction (b) of tartrates (2.3.1).
TESTS
F. (1R)-I-(3,4-dihydroxyphenyl)-2-(methylamino)
ethanesulfonic acid.
The solutionis not moreopalescent than reference
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph,<I
suspension II (2.2.1) and not more intensely coloured than
reference solution BY 5 (Z.2.2, MethodIf).
Dissolve0.5 g in water R and dilute to 10 mL with the same
solvent. Examine the solution immediately.
Adrenaline Acid Tartrate I Related substances
Epinephrine Acid Tartrate Liquid chromatography (Z.2.29). Prepare the solutions protected
from h"ghc
(Adrenaline Tartrate, Ph. Bur. monograph 0254)
Solvenl mixture A Dissolve 5.0 g of potassium dihydrogen
H O i f-, H OH~, H OH phosphote R and then 2.6 g of sodium oaonesalfonase R in
water for chromatagraphy R, and dilute to 1000 mL with the
I ~ CH, • Ho,CXyco,H
same solvent (it is usually necessary to stirfor at least30 min
HO -'" H OH
to achieve complete dissolution). Adjust to pH 2.8 with
pho,phoric acidR.
C"H,,NO, 333.3 51-42-3 Solvem mixture B acetonitrile Rt, solvent mixture A
(130:870 VIV).
Action and use
Adrenoceptor agonist. Test solution Dissolve75 mg of the substance to be
examined in 5 mL of O. I M hydroch/ori< acid and dilute to
Preparations
50 mL with solventmixture B.
Adrenaline InjectionlEpinephrine Injection
Reference solution (a) Dilute 1.0 mL of the test solutionto
Dilute Adrenaline Injection (I in 10,OOO)lDilute Epinephrine
100.0 mL with solvent mixrure B. Dilute 1.0 mL of this
Injection (I in 10,000)
solution to 10.0 rnL withsolvent mixture B.
Adrenaline SolutionlEpinepbrine Solution
Reference solution (b) Dissolve 1.5 mg of noradrenaline
Adrenaline and CocaineIntranasal Solution tartrate CRS (impurity B) and 1.5 mg of adrenalone
Bupivacaine and Adrenaline InjectionIBupivacaine and hydrochloride R (impurity C) in solvent mixture B, add
Epinephrine Injection 1.0 mL of the test solution and dilute to 100.0 mL with
Lidocaine and Adrenaline Injection/Lidocaine and solvent mixture B.
Epinephrine Injection
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2022 Adrenaline Acid Tartrate 1-83
Reference solution (c) Dissolve the contents of a vial of - disregard limit: 0.5 times the area of the principal peak in
adrenaline impurity mixture CRS (impurities D and E) in the chromatogram obtained with reference solution (a)
0.1 mL of 0.1 M hydrochloric acid and 0.9 mL of solvent (0.05 per cent).
mixture B. Los. on drying (2.2.32)
Reference solution (d) Dissolve 7.5 mg of adrenaline tartrate Maximum 005 per cent, determined on 10000 g by drying in
wirh impurity A CRS in 0.5 mL of 0.1 M hydrochlon'c acid and vacuo for 18 h.
dilute [0 5.0 mL with solvent mixture B. Sulfated ash (2.4.14)
Blank solution 0.1 M hydrochloric acid, solvent mixture B Maximum 0.1 per cent, determined on 1.0 g.
(1:9 VIII).
ASSAY
Column:
Dissolve 0.300 g in 50 mL of anhydrous acetic acidR, heating
~ size: 1= 0.10 m, 0 = 4.6 mm; gently if necessary. Titrate with 001 M perchkmc add until a
- stationary phase: end-capped octadecylsj/yl silica gelfor
bluish-green colour is obtained, using 0.1 mL of crystal violet
chromatography R (3 urn); solution R as indicator.
- temperature: SO "C.
I mL of 0.1 M perch/oric acid is equivalent to 33.33 mg
Mobilephase: of C 13H L9NOg •
- mobile phase A: acetonitrile Rl, solvent mixture A
(5:95 VIII); STORAGE
- mobile phase B: acetonitrile RJ, solvent mixture A In an airtight container, or preferably in a sealed tube under
(45:55 VIII); vacuum or under an inert gas, protected from light.
IMPURITIES
(min) (per cent VIJJ)
Specified impunOties A, B, C, D, E.
(per cent VIJ')
0-15 92 ..... 50 8 ..... 50
A. unknown structure,
15·20 50 ..... 92 50 -> 8 H OH
20 - 25 92
• HO~NH'
Flow rate 2.0 mllrnin. HOV
B. (IR)-2-amino-I-(3,4-dihydroxyphenyl)elhanol
Injection 20 111-. (noradrenaline),
Identification of impurities . Use the chromatogram supplied
with adrenaline impun°ty mixture CRS and the chromatogram o
obtained with reference solution (c) to identify the peaks due
to impurities D and Ej use the chromatogram supplied with Hoif~'
I CH,
adrenaline tartrate with impuni;y A CRS and the chromatogram HO
obtained with reference solution (d) to identify the peak due
to impurity A. C. 1-(3,4-dihydroxyphenyl)-2-(methylamino)ethanone
Relative retention With reference to adrenaline (retention (adrenalone),
=
time about 4 min): impurity B about 0.8j =
impurity C = about 1.3; impurity A = about 302j
impurity D =
about 3.3; impurity E about 3.7. =
System suitabiUty Reference solution (b):
impurity B and adrenaline.
Limits:
- correction factors: for the calculation of content, multiply D. 4-[(IR)-2-(benzylmethylamino)-I-hydroxyelhyl]benzene-
the peak areas of the following impurities by the 1,2-diol,
corresponding correction factor: impurity D = 0.7;
impurity E = 0.6;
- impun°ty A: not more than 3 times the area of the
reference solution (a) (0.3 per cent);
- impurities B, C: for each impurity, not more than twice the
E. 2-(benzyhnethylamino)-I-(3,4-dihydroxyphenyl)elhanone.
- impurities D, E: for each impurity, not more than the area
_ _ _ _ _ _ _ _ _~ -_ _- _ - _ -_ _ PhE"
.In the chromatogram obtained with reference solution (a)
(0.6 per cent);
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1-84 Agar 2022
*** solution add 5 mL of pictic acidsolution R. No turbidity

Agar
** ** appears within lO min.
(Ph. Eur. monograph 03/0)
***** Loss on drying (2.2.32)
Maximum 20.0 per cent, determined on 1.000 g of the
Action and use
powdered herbal drug (355) (2.9. /2) by drying in an oven at
Excipient.
105 "C.
PhE" _
Total ash (2.4./6)
DEFINITION Maximwn 5.0 per cent.
Polysaccharides from various species of Rhodophyceae Microbial contamination
mainly belonging to the genus Gelidium. It is prepared by TAMC: acceptance criterion 10' CFUlg (2.6./2).
treating the algae with boiling water; the extract is filtered TYMC; acceptance criterion 102 CFUlg (2.6./2).
whilsthot, concentrated and dried. Absence of Escherichia coli (2.6.13).
CHARACTERS Absence of Salmonella (2.6. /3).
Appearance
LABELLING
Powderor crumpled strips 2-5 mm wide or sometimes flakes,
The label states the swelling index.
colourless or pale yellow, translucent) somewhat tough and
~ ~ ~ PhE<I
difficult to break, becomingmore brittle on drying.
Mucilaginous taste.
IDENTIFICATION
***
A. Examine undera microscope. When mounted in 0.005 M
iodine, the strips or flakes are partly stained brownish-violet.
Medical Air •* **
Magnified 100 times, they show the following diagnostic (JUedicinal Air, Ph. Eur. monograph /238)
*****
characters: numerous minute, colourless, ovoid or rounded
grains on an amorphous background; occasional brown, U7hen kledical Air is intended for use in a room in which
round or ovoid spores with a reticulated surface) measuring magnetic resonanu imaging (kIRl) is being performed, the
up to 60 11m, may be present. Reduce to a powder, if cylinder andfittings should be madefrom suitable non-
necessary. The powder is yellowish-white. Examine undera ferromagnetic materials and labelled o=rding/y.
PhE" _
microscope using 0.005 .M iodine. The powder presents
angular fragments with numerous grains similar to those seen DEFINITION·
in the strips and flakes; some of the fragments are stained Compressed ambient air.
brownish-violet.
Content
B. Dissolve 0.1 g with heating in 50 mL of water R. Cool.
20.4 per cent VIV to 21.4 per cent VIV of oxygen (02 ) ,
To 1 mL of the mucilage carefully add 3 mL of water R so as
to fonn 2 separate layers. Add 0.1 mL of 0.05 M iodine. CHARACTERS
A dark brownish-violet colour appears at the interface. Mix. Appearance
The liquid becomes pale yellow. Colourless gas.
C. Heat 5 mL of the mucilage prepared for identification Solubility
test B on a water-hath with 0.5 mL of hydrochloric add R for At 20°C at a pressure of 101 kPa, 1 volumedissolves in
30 min. Add 1 mL of barium chloride solution R/. A white about 50 volumes of water.
turbidity develops within 30 min.
PRODUCTION
D. Heat 0.5 g with 50 mL of water R on a water-bath until Carbon dioxide
dissolved. Only a few fragments remain insoluble. During Maximum 500 ppm VIV, determined using an infrared
cooling, the solutiongels between 35°C and 30 °C. Heat the analyser (2.5.24).
gel thus obtainedon a water-bath; it does not liquefy below
Gas to be examined Filter the substance to be examined to
80 "C.
avoid stray light phenomena.
TESTS Reference gas (a) Use a mixture of21 per cent VIV of
Swelling index (2.8.4) oxygen Rand 79 per cent VIV of nitrogen Rl, containing less
Minimum 10 and within 10 per cent of the value stated on than I ppm VIV of corban dioxide R/.
the label, determined on the powdered herbal drug (355)
Reference gas (b) Use a mixture of 21 per cent VIV of
(2.9.12).
oxygen Rand 79 per cent VIV of nitrogen RJ, containing
Insoluble matter 500 ppm VIV of corban dioxide R/.
Maximum 1.0 per cent. Calibrate the apparatus and set the sensitivity using reference
To 5.00 g of the powdered herbal drug (355) (2.9./2) add gases (a) and (b). Measure the contentof carbon dioxide in
100 mL of waterRand 14 mL of dIlute hydrochkmc acidR. the gas to be examined.
Boil gentlyfor 15 min with frequent stirring. Filter the hot Carbon monoxide
liquid through a tared, sintered-glass filter (160) (2./.2), rinse
Maximum 5 ppm VIV, determined usingan infrared analyser
the filter with hot waterR and dry at 100-105 "C.
(2.5.25).
The residue weighs a maximum of 50 mg.
Gas to be examined Filter the substance to be examined to
Gelatin avoid stray light phenomena.
To 1.00 g add 100 mL of water R and heat on a water-bath
Reference gas (a) Use a mixture of 21 per cent VIV of
until dissolved. Allow to cool to 50°C. To 5 mL of this
oxygen Rand 79 per cent VIVof nitrogen RJ, containing less
than 1 ppm VIV of carbon monoxide R.
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2022 Air 1-85
0( Chopper
Sample in Sample out
M t t
I~_I
D~ •>•
UV Source i Filletonm
Collimator
0( Filler 350 nm
t
Photomultiplier
t
Amplifier
Figure 1238.-1. - Ut/fiuorescence analYser
Reference gas (b) Use a mixture of 21 per cent VIV of Gas to be examined The substance to be examined.
oxygen Rand 79 per cent VIV of nitrogen Rl, containing Reference gas (a) Use a mixture of 21 per cent VIVof
5 ppm VIV of carbon monoxide R. o.\)'gen Rand 79 per cent VIV of nitrogen R1, containing less
Calibrate me apparatus and set the sensitivity using reference than 0.05 ppm VIV of nitrogen monoxide and nitrogen
gases (a) and (b). Measure the content of carbon monoxide dioxide.
in the gas to be examined. Reference gas (b) Use a mixture of 2 ppm VIV of nitrogen
Sulfur dioxide monoxide R in nitrogen Rt,
Maximum 1 ppm VIr;, determined using an ultraviolet Calibrate the apparatus and set the sensitivity using reference
fluorescence analyser (Figure 1238.-1). gases (a) and (b). Measure the content of nitrogen monoxide
The apparatus consists of the following: and nitrogen dioxide in the gas (0 be examined.
- a system generating ultraviolet radiation with a wavelength Water
0£210 om, made up of an ultraviolet lamp, a collimator, Maximum 67 ppm VIV, determined using an electrolytic
and a selective filter; the beam is blocked periodically by a hygrometer (2.5.28), except where the competent authority
chopper rotating at high speeds; decides that the following limit applies to medicinal air
- a reaction chamber, through which flows the gas to be generated on-site and distributed in pipe-line systems
examined; operating at a pressure not greater than lObar and a
- a system that detects radiation emitted at a wavelength of temperature not less than 5°C: maximum 870 ppm VIY,
350 om, made up of a selective filter, a photomultiplier determined using an electrolytic hygrometer (2.5.28).
tube and an amplifier.
Assay
Gas to be examined Filter the substance (0 be examined. Determine the concentration of oxygen in air using a
Reference gas (a) Use a mixture of 21 per cent VIV of paramagnetic analyser (2.5.27).
oxygen Rand 79 per cent VIVof nitrogen Rt.
IDENTIFICATION
Reference gas (b) Use a mixture of 21 per cent VIVof Firstidentification: C.
oxygen Rand 79 per cent VIV of nitrogen R1, containing
Second identification: A, B.
0.5 ppm VIV to 2 ppm VIV of sulfurdioxide Rt,
A. In a conical flask containing me substance co be
Calibrate the apparatus and set the sensitivity using reference
examined, place a glowing wood splinter. The splinter
gases (a) and (b). Measure the content of sulfur dioxide in
remains glowing.
the gas to be examined.
B. Use a gas burette (Figure 1238.-2) of 25 mL capacity in
Oil the form of a chamber in the middle of which is a tube
Maximum 0.1 mglm3, determined using an oil detector tube
graduated in 0.2 per cent between 19.0 per cent and
(2.1.6), when an oil-lubricated compressor is used for the
23.0 per cent, and isolated at each end by a tap with a
production. conical barrel. The lower tap is joined to a tube with an
Nitrogen monoxide and nitrogen dioxide olive-shaped nozzle and is used to introduce the gas into the
Maximum 2 ppm VIV in total) determined using a apparatus. A cylindrical funnel above the upper tap is used to
chemiluminescence analyser (2.5.26). introduce the absorbent solution. Wash the burette with
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1-86 Air 2022
C. It complies with the limits of the assay.

TESTS
Carbon dioxide
Maximum 500 ppm V/V, determined using a carbon dioxide
detector rube (2./.6).
Sulfur dioxide
1.---25mm Maximum 1 ppm V/~ determined using a sulfur dioxide
detector rube (2./.6).
Oil
Maximum 0.1 mg/m), determined using an oil detector tube
(2.1.6)J when an oil-lubricated compressor is used for the
production.
Nitrogen monoxide and nitrogen dioxide
.Maximum 2 ppm VIV, determined using a nitrogen
monoxide and nitrogen dioxide detector tube (2.1.6).
Carbon monoxide
Maximwn 5 ppm VIVJ determined using a carbon monoxide
detector rube (2.1.6).
370mm Water vapour
Maximum 67 ppm VIVJ determined using a water vapour
detector tube (2.1.6)J except where the competent authority
decides that the following limit applies to medicinal air
23.0%
generated on-site and distributed in pipe-line systems
operating at a pressure not greater than 10 bar and a
temperature not less than 5 °C: maximum 870 ppm VIV,
determined using a water vapour detector rube (2.1.6).
210mm
STORAGE
21.0%
As a gas, in suitable containers complying with the legal
regulations or as a gas supplied by a pipe network.
--~,*"f-- 8 mm
LABELLING
Where applicable, the label states the production method, as
regards to the use of an oil - lubricated compression.
IMPURITIES
A. CO2 : carbon dioxide,
oOIf--25mm
B. SO" sulfur dioxide,
C. NO: nitrogen monoxide, _
D. N02 : nitrogen dioxide,
E. oil,
F. CO: carbon monoxide,
G. H 20: water.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph'Id
Figure 1238.-2. - Gas burette

Synthetic Air
water R and dry. Open the 2 taps. Connect the nozzle to the (Syntheric Medicinal A,r, Ph. Bur. monograph /684)
source of the gas to he examined and set the flow rate to
1 Umin. Flush the burette by passing the gas to be examined When Synthaic Air is intended for use in a room in which
through it for 1 min. Close the lower tap of the burette and magnetic resonance imaging (MRl) is being peifonnedJ the
immediately afterwards the upper tap. Rapidly disconnect the cylinder andjiuings should be madefrom suitable ntm-
burette from the source of the gas to he examined. Rapidly ferromagnetic materials and labelled a"",dingly.
give a half tum to the upper tap to eliminate any excess Ph 'II _
pressure in the burette. Keeping the burette vertical, fill the
DEFINITION
funnel with a freshly prepared mixture of 21 mL of. 560 gIL
Mixture of Nitrogen (/247) and Oxygen (04/7).
solution of potassium hydroxide Rand 130 mL of a 200 gIL
solution of sodium dithioniee R. Open the upper tap slowly. Content
The solution absorbs the oxygen and enters the burette. 95.0 per cent to 105.0 per cent of the nominal value which is
Allow to stand for 10 min without shaking. Read the level of between 21.0 per cent VIV to 22.5 per cenr VIV of oxygen
the liquid meniscus on the graduated part of the burette. (0,).
This figure represents the percentage VIV of oxygen. CHARACTERS
The value read is 20.4 to 21.4. Colourless and odourless gas.
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2022 Alanine 1-87
B. Use a gas burette (Figure 1684.-1) of25 mLcapacity in

A
the form of a chamber, in me middle of which is a tube
graduated in 0.2 per cent between 19.0 per cent and
23.0 per cent, and isolated at each end by a tap with a
conical barrel. The lower tap is joined to a tube with an
olive-shaped nozzle and is used to introduce the gas into the
apparatus. A cylindrical funnel above the upper tap is used to
k---25mm introduce the absorbent solution. Wash the burette with
water R and dry. Open both taps. Connect the nozzle £0 the
source of the substance £0 be examined and set the flow rate
to 1 Umin. Flush the burette by passing the substance to be
examined through it for 1 min. Close the lower tap of the
burette and immediately afterwards the upper tap. Rapidly
disconnect the burette from the source of the substance to be
examined. Rapidly give a half turn. of the upper tap to
eliminate any excess pressure in the burette. Keeping the
burette vertical, fill the funnel with a freshly prepared mixture
of 21 mL of a 560 gIL solution of potassium hydroxide Rand
130 mL of a 200 gIL solution of sodium di,hion;ce R. Open
the upper tap slowly. The solution absorbs the oxygen and
370mm enters the burette. Allow (0 stand for 10 min without
shaking. Read the level of the liquid meniscus on the
graduated part of the burette. This figure represents the
percentage VIV of oxygen. The value read is 95.0 per cent to
23.0%
105.0 per cent of the nominal value.
C. It complies with the limits of the assay.
TESTS
210mm Water vapour
21.0% Maximum 67 ppm VIV', determined using a water vapour
detector tube (2.1.6).
--~IttlI-~--8mm STORAGE
As a compressed gas in suitable containers complying with
the legal regulations or as a compressed gas supplied by a
pipe network, after mixing of the components.
19.0%
LABELLING
The label states the nominal content of O2 in per cent VIV.
+--25mm
IMPURITIES
A. H 20: water.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE"
Alanine
Figure 1684.-1.- Gas burette
SolubUlty
At a temperature of 20°C and a pressure of 101 kPa, C,H,NO, 89.1 56-41-7
1 volume dissolves in about 50 volumes of water.
PRODUCTION Action and use
Water (2.5.28)
Amino acid.
Maximum 67 ppm V/V. PIlE" _
Assay (2.5.27)
DEFINITION
Carry out the determination of oxygen in gases.
(2S)-2-Aminopropanoic acid.
IDENTIFICATION Content
Fi"t identification: C. 98.5 per cent to 101.0 per cent (dried substance).
CHARACTERS
A. In a conical flask containing the substance to be Appearance
examined, place a glowing splinter of wood. The splinter White or almost white, crystalline powder or colourless
remains glowing. crystals.
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1-88 Alanine 2022
Solubility Reference solution (a) Dilute 1.0 mL of the test solution to

Freely soluble in water, veryslightlysoluble in ethanol 100.0 mL with solutionA. Dilute 2.0 mL of this solution to
(96 per cent). 10.0 mL with solntion A.
IDENTIFICATION Reference solution (1)) Dissolve 30.0 mg of proline R in
First identification: A.. B. solution A and dilute to 100.0 mL with solution A. Dilute
1.0 mL of the solution to 250.0 mL with solution A.
Second identification: A, C, D.
Reference solution (c) Dilute 6.0 rnL of ammonium standard
solution (100 ppm NHJ R to 50.0 mL with solution A. Dilute
B. Infrared absorption spectrophotometry (2.2,24). 1.0 mL of this solution to 100.0 mL with solution A.
Comparison alanine GRS. Reference ,olution (d) Dissolve 30 mg of isoleucine Rand
C. Thin-layer chromatography (2.2.27). 30 mg of leucine R in solution A and dilute to 50.0 mL with
Testsolution Dissolve 10 mg of me substance £0 be solntion A. Dilute 1.0 mL of the solution to 200.0 mL with
examined in water Rand dilute to 50 mL with the same solution A.
solvent. Blank solution Solution A.
Reference soluuon Dissolve 10 mg of alanine CRS in water R Inject suitable, equal amounts of the test, blank andreference
and dilute to 50 mL with the same solvent. solutions into the amino acid analyser. Run a program
Plait TLC silica gelplait R. suitable for the determination of physiological amino acids.
Mobile phase glacial acetic acid R, water R, butanol R Systtm suitability Reference solution (d):
(20:20:60 VIV/V). - resolution: minimum 1.5 between the peaks due to
Applica.ion 5 ~L. isoleucine and leucine.
Development Over 2/3 of the plate. Calculation of percentage contents:
- for any ninhydrin-positive substance detected at 570 run,
Drying In air. use the concentration of alanine in reference solution (e),
Detection Spray with ninhydrin solution R and heat at 105 °C - for any ninhydrin-positive substance detected at 440 nm,
for 15 min. use the concentration of proline in reference solution (b);
Results The principal spot in the chromatogram obtained if a peak is above the reporting threshold at both
with the test solution is similar in position, colour and size to wavelengths, use the result obtained at 570 nm for
the principal spot in the chromatogram obtained with the quantification.
reference solution. Limils:
D. Dissolve 0.5 g in a mixture of 0.25 mL of hydrodlloric - allYninhydrin-positive subseana: for each impurity,
acidRI, 0.5 mL of a 100 gIL solution of sodium nUn'It R and maximum 0.10 per cent;
I mL of waterR. Shake; gas is given off. Add 2 mL of dilult - total: maximum 0.5 per cent;
sodium hydroxide solution R, foUowed by 0.25 mL of iodinattd - reporting threshold: 0.05 per cent.
potaSsium iodide solution R. After about 30 min, a yellow Chlorides (2.4.4)
precipitate is formed. Maximum 200 ppm.
TESTS Dilute 5 mL of solution S to 15 mL with waterR.
Solution S Sulfates (2.4.13)
Dissolve 2.5 g in distilled water R and dilute to 50 mL with Maximum 300 ppm.
the samesolvent.
Dilute 10 mL of solution S to 15 mL with distilled waltr R.
The solution is clear (2.2.1) and not more intensely coloured Ammonium
Amino acid analysis (2.2.56) as described in the test for
than reference solution BY. (2.2.2, Method II).
ninhydrin-positive substances with the following
Dilute 10 mL of solution S to 20 mL with waterR. modifications.
Specific optical rotation (2.2.7) Injection Test solution, reference solution (e) and blank
+ 13.5 to + 15.5 (dried substance). solution.
Dissolve 2.50 g in hydrochloric acid RJ and dilute to 25.0 mL Limit:
with the same acid. - ammonium' at 570 nm: not more than the area of the
Ninhydrin-positive substances corresponding peak in the chromatogram obtained with
Amino acid analysis (2.2.56). For analysis, use Method 1. reference solution (c) (0.02 per cent), taking into account
The concentrations of the test solution and the reference the peak due to arrunoniwn in the chromatogram
solutions may he adapted according to the sensitivity of the obtained with the blank solution.
equipment used. The concentrations of all solutions are Iron (2.4.9)
adjusted so that the system suitability requirements described Maximum 10 ppm.
in general chapter 2.2.46 arefulfilled, keeping the ratios of In a separating funnel, dissolve 1.0 g in 10 mL of dilult
concentrations between all solutions as described. hydrochloric acid R. Shake with 3 quantities, each of 10 rnl.,
Sohuion A dilult hydrochloric acidRJ or a sample preparation of methyl isobutyl ketone RI, shaking for 3 min each time.
buffer suitable for the apparatus used. To the combined organic layers add 10 rnL of water R and
Test solution Dissolve 30.0 mg of the substance to be shake for 3 min. Use the aqueous layer.
examined in solution A and dilute to 50.0 mL with Loss on drying (2.2.32)
solution A. Maximum 0.5 per cent, determined on 1.000 g by drying in
an oven at 105 "C.
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2022 Albendazole 1-89
Sulfated ash (2.4.14) Solubility

Maximum 0.1 per cent, determined on 1.0 g. Practically insoluble in water, freelysoluble in anhydrous
formic acid, vecyslightly soluble in methylene chloride,
ASSAY
practically insoluble in ethanol (96 per cent).
Dissolve 80.0 mg in 3 mL of anhydrous formic add R.
Add 30 mL of anhydrous acetic acid R. Titrate with 0.1 M It shows polymorphism (5.9).
perchloric acid, determining Ute end-pointpotentiometrically IDENTIFICATION
(2.2. 2/J). Infrared absorption spectrophotometry (2.2.24).
I mL of 0.1 M perch/oric acid is equivalent to 8.91 mg of Comparison albendazole CRS.
C,H,N02 . If the spectra obtained in the solid state show differences,
STORAGE dissolve the substance to he examined and the reference
Protected from light. substance separately in methylene chloride R, evaporate to
dryness and record new spectra using the residues.
IMPURITIES
Otherdetectable impurities (thefollowing substances would, '1 TESTS
present at a sufficient leuel, be detected by oneor other of the tests Appearance of solution
in the monograph. They are limited by the general acceptance The solutionis clear (2.2.1) and not more intensely coloured
cdtenonfor otherhmspecified impurities and/or by thegeneral than reference solutionBY6 (2.2.2, Method If).
monograph Subseances for pharmaceutical use (2034). It is Dissolve 0.10 g in a mixture of 10 volumesof anhydrous
therefore not necessary to identify these impurities for formic acid Rand 90 volumes of methylene chloride Rand
demonstration of compliance. See also 5.10. Control of impurities dilute to J0 mL with the same mixture of solvents.
in substances for pharmaceutical use) A.. B. Related substances
H Nt<,
HD,C 0 CD,H
Soioem mixsure su/furic acidR, methanol R (1 :99 VIV).
A. (2S)-2-aminobutanedioic acid (aspartic acid), examined in 5 mL of the solvent mixture and immediately
dilute to 50.0 mL with the mobile phase.
H Nt<,
Reference solution (aJ Dilute 1.0 mL of the test solution to
HO:!C~C0:2H 100.0 mL with the mobile phase. Dilute 1.0 mL of this
B. (2S)-2-aminopentanedioic acid (glutamic acid). Reference sdunon (b) Dissolve 5 mg of albendazole for system
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE.. suitability CRS (containing impurities B, C, B, F and H) in
1 mL of the solvent mixture and dilute [0 10 mL with the
mobile phase.
Reference solution (c) Dilute 1 mL of the solvent mixture to
Albendazole 10 mL with the mobile phase. Use 1 mL of this solution to
dissolve the contents of a vial of albendazole
(Ph. Eur. monograph 1386) impurity mixture CRS (containing impurities A and D).
Column:
- size: / = 0.25 m, 0 = 4.6 rnm;
- stationary phase: end-capped octade<ylsilyl silica gelfor
chromatography R (5 urn).
MobI7e phase 1.67 gIL solution of ammonium dihydrogen
phosphate R, methanol R (30:70 VIV).
265.3 54965-21-8
Flow rat< 0.7 mUmin.
Action and use Detection Spectrophotometer at 254 nm.
Benzimidazole antihelminthic. Injection 20 IJL.
Preparations Run time Twice the retention time of albendazole.
Albendazole Oral Suspension Identification of impurilies Use the chromatogram supplied
Albendazole Oral Suspension with Minerals with albendazole impurity mixture CRS and the chromatogram
obtained with reference solution (c) [Q identify the peaks due
PhE.. _
to impurities A and D; use the chromatogram supplied with
DEFINITION albendazole for system suitability CRS and the chromatogram
Methyl N-[5-(propylsulfanyl)-IH-benzimidazol-2-yl] obtained with reference solution (b) to identify the peaks due
carbamate. to impurities B + C, E, F and H.
Content Relative retention With reference to albendazole (retention
99.0 per cent to 101.0 per cent (dried substance). = =
time about I I min): impurity D about 0.35;
impurities Band C = about 0.40; impurity E = about 0.45j
CHARACTERS impurity A = about 0.48; impurity F == about 0.57;
Appearance impurity H = about 0.66.
White or slightly yellowish powder. System suitability Reference solution (b):
- resolution: minimum 1.5 between the peaks due [0
impurities B + C and E.
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1-90 Albendazole 2022

- correction factors: multiply the peak areas of the following
impurities by the corresponding correction factor:
impurity A::::;; 1.7; impurities Band C == 1.4;
impurity D = 1.9; impurity E = 1.4;
- for each impurity, use the concentration of albendazole in
C. methyl N-[5-(PropylsuLfonyl)-IH-benzimidazol-2-
yljcarbamate,
Limits:
- impmiey H: maximum 0.6 per cent;
- impurity F: maximwn 05 per cent;
- impurity A: maximum 0.4 per cent;
- sum oj impurities Band C: maximum 0.4 per cent;
- impuniy E: maximum 0.3 per cent;
- impurity D: maximum 0.2 per cent;
- unspecified impurities: for each impurity J maximwn D. (2-amino-lH-benzimidazol-5-yl)propyl-)..'-suLfanedione,
0.10 per cent;
er
- total: maximum 1.3 per cent; H 0
N }-OCH,
I N
r NH
an oven at 105°C for 4 h. E. methyl N-(IH-benzimidazol-2-yl)carbamate,
ASSAY
In order to avoid overheating dunng the (imllion, mix thoroughly
throughout and SlOP the titration immediately afterthe end-point
has been reached. F. methyl N-[5-(methylsuLfanyl)-IH-benzimidazol-2-
Dissolve 0.250 g in 3 mL of anhydrous fonni' acidR and add yljcarbamate,
40 mL of anhydrous acetic acidR. Titrate with 0.1 M
perchknic acid) determining-the end-point potentiometrically
(2.2.20).
of ClzH15N30ZS.
STORAGE G. methyl N-(5-<:hloro-lH-benzimidazol-2-yl)carbamate,
IMPURITIES
Specified impun'ties A) B, C, D, E, F, H.
Other detectoble impurities (thefollowing substances would, if
present at a sufficient level) be detected by one or other of the tests
in the monograph. They arelimiud by thegeneral aaeptance H. methyl N-[5-[(2-methyl-4-oxopentan-2-yl)sulfanylj-lH-
criterion for otherlullspedjied impuniies andlor by the general benzhnidazol-2-yl]carbamate,
numcgroph Substances for pharmaceutical use (2034). It is
therefore not necessary to identify these impumies for o
DI r
demonstration of compliance. See also5.10. Control 0/impurities ~ }-OCH,
in substances for pharmaceutical use) 0, I) J, K, L
HC_ »<:
3 .............
~
~o............ N
NH
1. methyl N-(5-propoxy-lH-benzimidazol-2-yl)carbamate,
H 0
ClVN }-OCH,
A. 5-(propylsuLfanyl)-IH-benzimidazol-2-amine,
~
I N
r NH
o
("'y ~ }- OCH, CI
H'C~sAJ-.N}-NH J. methyl N-(4,6-dichloro-lH-benzimidazol-2-yl)carbamate,

o"
B. methyl N-[5-(propylsulfinyl)-IH-benzimidazol-2-
yl]carbamate,
K. methyl N-[5-(butylsuifanyl)-IH-benzimidazol-2-ylJ
carbamate)
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2022 Alcuronium Chloride 1-91
Application 10 J.lL.
Deodopment Overa path of 15 em.
Drying In air for 10 min.
Detection Spray with 0.1 M ammonium and cerium nitrate.
L. methyl N-[5-[(propan-2-yl)sulfanyl]-1 H-benzimidazol-2- Results The principal spot in the chromatogram obtained
yl]carbamate. with the test solutionis similar in position, colour and size to
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'/>81 the principal spot in the chromatogram obtainedwith the
reference solution.
C. It gives reaction (a) of chlorides (2.1.1).
TESTS
Alcuronium Chloride Solution S
(ph. Eur. monograph 1285) Dissolve 0.250 g in carbon dioxide-free water R and dilute to
Solution S is clear (2.2.1) and not more intenselycoloured
chan reference solution Y61 BY6 or B 6 (2.2.2, klethod l).
Acidity or a1ka1lnlty
z cr To 10 mL of solution S add 0.1 mL of methyl redsolution R
and 0.2 mL of 0.01 M hydrochloric acid. The solution is red.
Add 0.4 mL of 0.01 M sodium hydroxide. The solution is
yellow.
-430 to -451 (anhydrous substance), determined on
738 1518()'01-7 solution S.
Propan-z-ol (2.4.24, System A)
Action and use Maximum 1.0 per cent.
Non-depolarizing neuromuscular blocker.
Related substances
--------c-------------
I'/>E"
DEFINITION Solventmixture Mix 100 mL of methanol R, 200 mL of
acetonitrile Rand 200 mL of a 6.82 gIL solution of potassium
(lR,3aS, lOS, lIaS,12R,14aS, 19a5,20bS,2IS,22aS,23E,26B)-
dihydrogen phosphate R. Dissolve 1.09 g of sodium
23,26-bis(2-Hydroxyethylidene)-1,12-bis(prop-2-enyl)-
laurylsu/fonate for chromatography R in the mixture and adjust
2,3,11,11 a,13,14,22,22a-octahydro-1 OH,2IH-I,21:10,12-
the apparent pH to 8.0 with a 100 gIL solution of sodium
diethano-19aH,20bH-[1 ,5Idiazocino[1 ,2,3-lm:5,6,7-1'm'l
hydroxide R.
dipyrrolo[2',3' -d:2'',3' I :d']dicarhazolediiuffi dichloride (4,4'-
didesmethyl-4,4'-bis(prop-2-enyl)toxiferin I dichloride). Test solution Dissolve 0.20 g of the substance to be
Content the solvent mixture.
98.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS 100.0 mL with the solvent mixture.
Appearance Reference solution (b) Dilute 4.0 mL of reference solution (a)
White or slightly greyish-white, crystalline powder. to 10.0 mL with the solvent mixture.
Solubility Reference solution (c) Dilute 1.0 mL of reference solution (a)
Freelysoluble in water and in methanol, soluble in ethanol to 10.0 mL with the solvent mixture.
(96 per cent), practically insoluble in cyclohexane.
Reference solution (d) To 5.0 mL of the test solution add
Carry out the identification, tests and assay as rapidlyas possible 5.0 mg of aHylstrychm'ne bromide CRS, dissolve in the solvent
avoidingexposure Ie actinic light. mixture and dilute to J00.0 mL with the solvent mixture.
IDENTIFICATION Column:
First identification: A, C. - size: I = 0.25 m, 0 = 4 mm;
Second identification: B, C. - stationary phase: octy/silyl silica gelfor chromatography R
(5 pm).
Comparison alcuronium chloride CRS. Mobile phase Mix 200 mL of methanol R, 400 mL of
acetonitrile Rand 400 mL of a 6.82 gIL solution of potassium
B. Thin-layer chromatography (2.2.27). dihydrogen phosphate R. Dissolve 2.18 g of sodium
Testsolution Dissolve 10 mg of the substance to be lautylsulfonate for chromawgraphy R in the mixture and adjust
examined in methanol R and dilute to 10 mL with the same the apparent pH to 5.4 with a 100 gIL solution of phosphoric
solvent. acidR.
Reference solution Dissolve 10 rng of akuroniumehloride CRS Flow rate 1.2 mUmin.
in methanol R and dilute to 10 mL with the same solvent. Detection Spectrophotometer at 254 om.
Plate TLC silica gelplateR. Injection I0 ~L.
Mobile phase Mix 15 volumes of a 58.4 gIL solution of Run time Twice the retention time of alcuronium.
sodium chloride R, 35 volumes of dilute ammonia R2 and
50 volumes of methanol R.
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1-92 Alfacalcidol 2022
System suitability Reference solution (d):

Alfacalcidol
N-allylstrychnine and alcuronium. (Ph. Eur. monograph 1286)
Limits:
- impurities A, B: for each impurity, not more than the area
CH,
reference solution (a) (0.5 per cent) and not more than
one of the peaks has an area greater than the area of the
- total: not more than twice the area of the principal peak in
(I per cent);
- disregard limit: the area of the principal peak in the
chromatogram obtained with reference solution (c)
(0.05 per cent). C"f4,02 400.6 41294-56-8
Water (2.5.12)
Action and use
Maximum 5.0 per cent, detenninedon 0.500 g. Vitamin D analogue.
PhE" _
ASSAY DEFINITION
Dissolve 0.300 g by stirring in 70 mL of acetic anhydride R (IS,3R,5Z, 7E)-9, 1u-Secocholesta-S,7, I O( 19)-triene-I,3-diol.
for 1 min. Titrate with 0.1 M perchfon'c acid until the colour Content
changes from violet-blue to greenish-blue, using 0.1 mL of 97.0 per cent to 102.0 per cent.
crystal viokt solution R as indicator. A reversible isomerisation (0 pre-alfacalcidol takes place in
I mL of 0.1 M perch/one <Kid is equivalent to 36.9 mg solution, depending on temperature and time. The activity is
of C..H,oCI,N,02. due to both compounds (see Assay).
STORAGE CHARACTERS
In an airtight container undernitrogen, protected from light, Appearance
at a temperature of 2 °C to 8°C. White or almostwhite crystals.
IMPURITIES Solubility
Specified impurities A, B. Practically insoluble in water, freely soluble in ethanol
(96 per cent), soluble in fatty oils.
It is sensitive to air, heat and light.
IDENTIFICATION
2Cf Comparison Ph. Eur. reference spearum of alfoualcidol.
related substances.
with the test solution is similar in retention time and size to
theprincipal peak in the chromatogram obtained with
A. (IR,3aS,9R,9aR,IOR,ll as,12R,14aS,19aS,20R,20aR,
20bS,21 R,22aS)-I,12-bis(prop-2-enyl)-
2,3,9a,11,11a)13) 14,19a,20a)21,22,22a-dodecahydro- TESTS
I OH,20bH-I,23:12,27-dimethano-9, I 0:20,21-bis Related substances
(epoxyprop[2]eno) -9H,20H- [1,5]diazocino[1,2 ,3-lm: 5,6,7- Liquid chromatography (2.2.29): use the nonnalisation
I'm']dipyrrolo[2',3'-d:2" ,3" :d']dicarbazolediium procedure. Cany out the test as rapidly as possible, avoiding
dichloride (4,4'-diallylcaracurin V dichloride), exposure to light and air, andprepare the solutWns immediately
before use.
examined in 25.0 mL of acetonitrile R and dilute to 50.0 rnL
with water R.
CI
Reference ,o{wion (a) Dissolve 5.0 mg of alfoualcido/ CRS in
25.0 mL of acetonitrile R and dilute to 50.0 mL with waterR.
Reference ,olution (b) Dilute 1.0 mL of reference solution (a)
(0 100.0 mL with a mixture of equal volumes of acetonitrile R
B. (4bS,7R, 7aS,8aR, 13R,13aR,13bS)-13-hydroxy-7-(prop-2- and water R. Dilute 1.0 mL of this solution to 20.0 mL with
enyl)-5,6,7a,8,8a,11,13,13a,13b,14-decahydro-7,9~ the same mixture of solvents.
methano-7H-oxepino[3,4-a]pyrrolo[2,3-d1carbazolium Reference sdudon (e) Dissolve 2 mg of alfacalcidol for system
chloride «4R, 17R)-4-allyl-17, 18-epoxy-17-hydroxy-19,20- ,,,,cabihiy A CRS (containing impurities A and D) in 10 mL
dldehydrocuranium chloride). of acetonitrile R and dilute to 20 mL with water R. Allow to
___________ PhE"
~ ~
stand at room temperature for about 2 h, ensuring tha t the
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2022 Alfacalcidol 1-93
signal-to-noise ratio of the peak due to pre-alfacalcidol is Otherdetectable impurities (thefollO'i.villg substances would, if
between 25 and 300. present at a sufficient level, be detected by oneor other of the tests
Reference solmion (d) Dissolve 2 mg of alfacalcidJJl for in the monograph. They are limited by thegeneral acceptance
impun"IY B identification CRS in 10 mL of acetonittiie Rand criterion for other/unspecified impuniies andlor by thegeneral
dilute to 20 mL with water R. monograph Substances for pharmaceutical use (2034). It is
Column: therefore not necessary to identify these impurities for
- size: 1= 0.15 m, 0 ;;;; 2.1 mmj demonstration ofcomplianu. See also 5.10. Control of impun'ties
- stationary phase: end-capped ethylene-bridged oetade<ylsilyl in substances for pharmaceutical use) C.
SIlica gelfor chromatography (hybridmateriaQ R (1. 7 ~m);
iHohiJe phase methanol R, waterfor chromatography R, CH,
acetonitrile R (15:17:68 VIVW).
FkJw race 0.3 mllmin.
Injection 5 JlL of the test solution and reference
solutions (b), (c) and (d).
Run time Twice the retention time of alfacalcidcl, HO-- ',OH
H H
Identification of imptmues Use the chromatogram supplied
with aJfacaicidol for system suitability A CRS and the A. (IS,3R,5E, 7E)-9,1 0-secocholesta-5,7,1 0(19)-triene-I,3-
chromatogram obtained with reference solution (c) to identify diol (trans-alfacalcidol),
the peaks due to impurities A and D and pre-alfacalcidol; use
the chromatogram supplied with alfaroltidol for impurity B
identification CRS and the chromatogram obtained with
reference solution (d) to identify the peak due to impurity B.
Relativeretention With reference to alfacalcidol (retention H,C
time = about 15 min): pre-alfacalcidol = about 0.91;
impurity A = about 0.94; impurity D = about 0.96;
impurity B = about l.l.
System suilabiJity Reference solution (c):
- resolution: minimum i.s between the peaks due to
impurity D and alfacalcidol;
- peak-to-vaUey ratio: minimwn 5.0 J where Hp height =
above the baseline of the peak due to impurity A and B. (I R,3R,5Z, 7E)-9,1 0-secocholesta-5,7,I0(19)-triene-I,3-
HI) = height above the baseline of the lowest point of the diol (l~-calcidol),
curve separating this peak from the peak due to pre-
alfacalcidol; minimum I.5 J where Hp = height above the
baseline of the peak due to impurity D and HI) height=
above the baseline of the lowest point of the curve eH,
separating this peak from the peak due to impurity A.
o
>--N
Limits:
- impurities A J B, D: for each impurity, maximum
0.5 per cent;
0-N)fN
, j
o eH,
- unspecified impurities: for each impurity, maximum
0.10 per cent; HO'" \ OH
- total: maximum 1.0 per cent; H H
chromatogram obtained with reference solution (b) C. 6~-[(3S,5R)-3,5-dihydroxy-2-methylcyclohex-l-en-I-yll
(0.05 per cent); disregard the peak due to pre-alfacalcidol. 17 ~- [(2R)-6-methylhep tan- 2-yl]- 2-phenyl-2,5, 1O-triaza-4-
nor-9~-estr-7 -ene-Lj-dione,
ASSAY
Liquid chromatography (1.1.19) as described in the test for D. unknown structure.
related substances with the following modification. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'hE«
Injection Test solution and reference solution (a).

Calculate the percentage content of C21~402 taking into
account the assigned content of olfacalcidol CRS and, if
present, the peak due to pre-alfacalcidol.
STORAGE
Under nitrogen, in an airtight container, protected from light,
at a temperature of 2 °C to 8°C.
The contents of an opened container are to be used
immediately.
IMPURITIES
Specified impurities A, B D.
J
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1-94 Alfadex 2022
Alfadex *** room temperature. Add 10 mL of ammonium molybdate

** **
**** *
reagent Rl and allow to stand for 15 min.
Alphacyclodextrin Reference solution Prepare a reference solution at the same
(Ph. Eur. monograph 1487) time and in the same manner as the test solution, using
I mL of a 0.02 gIL solution of glucose R.
OO)_(~qHX:'pHOH
Measure the absorbance (2.2.25) of the test solution and the
reference solution at the absorption maximum at 740 om
using warer R as the compensation liquid. The absorbance of
the test solution is not greater than that of the reference
\--no solution.
-o HO OH
O>---<OH HO
QH •
0
Light-absorbing Impurities
Examine solution S between 230 om and 750 om. Between
OO"".:Q~~
0 :
230 nrn and 350 nm, the absorbance (2.2.25) is not greater
than 0.10. Between 350 om and 750 om, the absorbance
(2.2.25) is not greater than 0.05.
Related substances
HQ OH
973 10016-2()-3
examined in water R with heating) cool and dilute to
Action and use 25.0 rnL with the same solvent.
Cyclodextran; carrier molecule for drug delivery systems. Test solution (b) Dilute 5.0 mL of test solution (a) to
Ph8l _ 50.0 mL with waw R.
Reference solution (a) Dissolve 25.0 mg of beuulex CRS
DEFINITION (impurity A), 25.0 mg of gammacydodextrin CRS
Cyclohexakis-( I ~ 4)-( u-n-glucopyranos yl) (impurity B) and 50.0 mg of alfadex CRS in waw R, then
(cyclomaltobexaose or c-cyclcdextrin). dilute to 50.0 mL with the same solvent.
Content Reference solution (b) Dilute 5.0 mL of reference solution (a)
97.0 per cent to 102.0 per cent (dried substance). to 50.0 mL with waw R.
CHARACTERS Reference solution (c) Dissolve 25.0 mg of alfadexCRS in
Appearance water R and dilute to 25.0 mL with the same solvent.
White or almost white, amorphous or crystalline powder. Column:
- size: 1= 0.25 TIl, 0 = 4.6 nun;
Solubility
- statianary phase: end-eapped octadecy/silyl silica gelfor
Freely soluble in water, slightly soluble in propylene glycol,
chroma/Ography R (10 urn).
practically insoluble in anhydrous ethanol and in methylene
chloride. Mobile phase methanol R, waw R (10:90 VIV).
FkJw rau 1.5 mUmin.
IDENTIFICATION
A. Specific optical rotation (see Tests). Detection Differential refractometer.
B. Examine the chromatograms obtained in the assay. Equilibration With the mobile phase for about 3 h.
Results The principal peak in the chromatogram obtained Injection 50 ~ of test solution (a) and reference
with test solution (b) is similar in retention time and size £0 solutions (a) and (b).
the principal peak in the chromatogram obtained with Run time 3.5 times the retention time of alfadex.
reference solution (c). Relative retention With reference to alfadex (retention
C. Dissolve 0.2 g in 2 mL of iodine solution R4 by warming time = about 10 min): impurity B = about 0.7;
on a water-bath, and allow to stand at room temperature; impurity A = about 2.2.
a yellowish-brown precipitate is fanned. System suitability Reference solution (a):
TESTS - resolution: minimum 1.5 between the peaks due to
impurity Band alfadex; if necessary, adjust the
Solution S
concentration of methanol in the mobile phase.
Dissolve 1.000 g in carbon dioxide-free water R and dilute to
100.0 mL with the same solvent. Limits:
- impurities A, B: for each impurity, not more than
0.5 times the area of the corresponding peak in the
Solution S is clear (2.2.1).
pH (2.2.3) (0.25 per cent);
5.0 to 8.0. - sum of impurities other than A and B: not more than
Mix I mL of a 223.6 gIL solution of potassium chloride Rand 0.5 times the area of the peak due to alfadex in the
30 mL of solution S. chromatogram obtained with reference solution (b)
Specific optical rotation (2.2.7) (0.5 per cent).
+ 147 to + 152 (dried substance), determined on solution S. Loss on drying (2.2.32)
Reducing sugars Maximum 11 per cent, determined on 1.000 g by drying in
Maximum 0.2 per cent. an oven at 120 °C for 2 h.
Test solmum To I mL of solution S add I mL of cupri- Sulfated ash (2.4.14)
lartan"c solution R4. Heat on a water-bath for 10 min, cool to Maximum 0.1 per cent, determined on 1.0 g.
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2022 Alfentanil Hydrochloride 1-95
**.
••* •**
ASSAY
Alfentanil Hydrochloride Hydrate
related substances with me foHowing modifications. Alfentanil Hydrochloride
*••
Injection Test solution (b) and reference solutions (a) (Ph. Bur. monograph 1062)
and (c).
System SUiMbi!ity:
- repeatability: maximum relative standard deviation of
2.0 per cent for the peak due to alfadex after 5 injections
of reference solution (a).
Calculate the percentage content of [C~1005]6 from the
assigned content of alfadex CRS.
STORAGE
In an airtight container.
LID>URITIES
453.0 (anhydrous substance)
Spedfied impuruies A, B.
Anhydrous a1fentanil hydrochloride 69049-06-5
HO.. «OH"O"yH,?OH Action and use
HO~
Ho·--\-d
O' 0
OH
a
HO HO p". OH
'0
OH
Opioid receptor agonist; analgesic.
PhE"
DEFINITION
_
6.~OH HO ;, N- [1- [2-(4-Ethyl-5-oxo-4,5-dihydro-l H-tetrazol-I-yl) ethylj-4-
S:-:~~"OH
(methoxymethyl)piperidin-4-ylj-N-phenylpropanamide
hydrochloride hydrate.
HO-\_{.
HO H
HO OH
OH
Content
It contains a variable quantity of water.
CHARACTERS
A. cycloheptakis-(1->4)-(a-D-glucopyranosyI) (betadex or Appearance
cyclcmaltoheptaose or Bcyclodextrin), White or almost white powder.
Solubility
HO H~H OH Freely soluble in water, in ethanol (96 per cent) and in
methanol.
HO.)--,··O-- 0) --O~Yr0H mp
Hox:r)---O~H HO Hor»
About 140°C, with decomposition.
··'? .OH It shows polymorphism (5.9).

OH O' IDENTIFICATION
. a HO A. Infrared absorption spectrophotometry (2.2.24).
~""JS~c):/~
Comparison alfentanil hydrochloride hydrate CRS.
oa dissolve the substance to be examined and the reference
HO
HO
H OH
OH
substance separately in methanol R, evaporate to dryness and
B. Dissolve 50 mg in a mixture of 0.4 mL of ammonia Rand
2 mL of water R. Mix, allow to stand for 5 min and filter.
B. cyclooctakis-(1->4)-(a-D-glucopyranosyl) Acidify the filtrate with dilute nitric acidR. It gives
(cyclornalroocraose or y-cyclodextrin). reaction (a) of chlorides (2.3.1).
_ _ _ _ _ _ _ _ _ _ _ _ _ _~ ~ I'1>E"
TESTS
Method II).
Dissolve 0.2 g in water R and dilute to 20 mL with the same
solvent.
Related substances
Liquid chromatography (2.2.29). Cany out she us, protected
from ligh,
examined in methanol R and dilute to 10.0 mL with the same
solvent.
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1-96 A1fentanil Hydrochloride 2022
Reference solution (aJ In order to prepare impurity E in situ, STORAGE

dissolve 10 mg of the substance to be examined in 10.0 mL Protected from light.
of dIlute hydrochlon'c acid R. Heat on a water-bath undera
IMPVRITIES
reflux condenser for 4 h. Neutralise with 10.0 mL of dilure
Specified impurities D.
sodium hydroxide SolUtWll R and evaporate to dryness on a
water-bath. Cool and take up me residue in 10 mL of Otherdetectable impurities (thefollowing substances «'QuldJ if
methanol R. Filter. present at a sufficient level, be detected by one or otherof the tests
in the monograph. They are limitedby thegeneral acceptance
Reference solUlion (b) Dissolve the contents of a vial of
cntetion for otherlunspec{fied impurities and/or by thegeneral
a/fentanil impuril)l D CRS in 1 mL of methanol R.
Reference solution (c) Dilute 1.0 mL of the test solution to therefore not necessary to identify these impuniies for
100.0 mL with methanol R. Dilute 1.0 mL of this solution to demonstration of compliance. See also 5.10. Control of impurities
10.0 mL with methanol R. in substances for pharmaceutical use) AJ B, C, E, F, G, H.
Blank solution methanol R.
Column:
- size: 1 = 0.1 m, 0 = 4.6 mm;
- stationary phase: end-copped O<tade<y/si!yl silica gelfor
chromatography R (3 pm).
Mobile phase:
- mobile phase A: 5 gIL solution of ammonium carbonate R in
a mixture of 10 volumes of tetrahydrofuran Rand
90 volumes of waterfor chromatography Rj A. (1,,4,)-1-[2-(4-ethyl-5-oxo-4,5-d ihydro-I H-terrazol-I-yl)
- mobile phase B: aceronitrile for chromatography R; ethyl)-4-(methoxymethyl)-4-(N-phenylpropanamido)
piperidine f-oxlde,
Time MobUe phase A MobUe phase B
(min) (per cent VM (per cent VII?
0- 15 90 ~ 40 10 ---> 60
15 - 20 40 60
20 - 25 40 ---> 90 60 ~ 10

Detection Spectrophotometer at 220 run. B. (I r,4r)-I-[2-(4-ethyl-5-oxo-4,5-dihydro-IH-tetrazol-I-yl)
lnjectum 10 pL. ethyl)-4-(methoxymethyl)-4-(N-phenylpropanamido)
Identification of impun·ties Use the chromatogram obtained piperidine f-oxlde,
with reference solution (a) to identify the peak due to
impurity H; use the chromatogram obtained with reference
solution (b) to identify the peak due to impurity D.
Relative retention With reference to alfentanil (retention
impurity E = about 0.9.
System suitab-iUty Reference solution (a):
impurity E and alfentanil.
C. N-[4-(methoxymethyl)piperidin-4-yl]-N-
phenylpropanamide,
Calculation ofpercentage contents:
- for each impurity, USe the concentration of alfentanil
hydrochloride hydrate in reference solution (c).
Limits:
- impurity D: maximum 0.2 per cent;
- unspecified impun·cies: for each impurity, maximum
0.10 per cent;
- toud: maximum 0.4 per cent;
- reporting threshold: 0.05 per cent. D. N- [1-[2-(-l-ethyl- 5-ox<f-4,5-dihydro-1 H-terrazol-l-yl)
Water (2.5.12) ethyl]-4-(methoxymethyl)piperidin-4-yl]-N-
3.0 percent to 4.0 per cent, determined on 0.500 g. phenylacetamide,
ASSAY
Dissolve 0.350 g in 50 mL of a mixture of 1 volume of
ethanol (96 perunV R and 4 volumes of woW' R and add
5.0 mL of 0.01 M hydrochloric acid. Titrate with 0.1 M
sodium hydroxide, determining the end-point
potentiometrically (2.2. 20). Read the volume added between
the 2 pointsof inflexion.
1 mL of 0.1 M sodium hydroxide is equivalent to 45.30 mg of E. l-ethyl-4-[2-[4-(methoxymethyl)-4-
C21H33CIN603' (phenylamino)piperidin-I-yl)ethyl]-1,4-dihydro-5H-
terrazol-S-one,
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2022 Alfuzosin Hydrochloride 1-97
Solubility
Freely soluble in water, sparingly soluble in ethanol
(96 per cent), practically insoluble in methylene chloride.
IDENTIFICATION
Comparison olfuzosin hydrochloride CRS.
F. N-[ 1-(2-hydroxyethyl)-4-(methoxymethyI)piperidin-4-yl)- B. It gives reaction (a) of chlorides (2.3.1).
N-phenylpropanamide, TESTS
pH (2.2.3)
4.0 to 5.5.
25.0 mL with the same solvent. Use a freshly prepared
solution.
Related substances
Liquid chromatography (2.2.2'l).
examined in the mobile phase and dilute to 100.0 mL with
G. N-[I-[2-(4-ethyl-5-oxo-4,5-dihydro-1H-tetrazol-l-y1) the mobilephase.
ethyl]-4-[(propanoyloxy)methyl)piperidin-4-yl]-N-
Reference solution (aJ Dilute 1.0 mL of the test solutionto
phenylpropanamide,
100.0 mL with the mobile phase. Dilute 1.0 mL of this
solution to 10.0 mL with the mobilephase.
Reference ,0IUlUm (b) Dissolve 4 mg of alfuzo,in for system
,uitability A CRS (containing impurities B, F and G) in the
mobilephase and dilute to 10.0 mL with the mobile phase.
Reference solution (c) Dissolve 4 mg of alfuzosin for peak
identification CRS (containing impurity D) in the mobile
phase and dilute to 10.0 mL with the mobile phase.
H. N-[I-[2-(4-ethyl-5-oxo-4,5-dihydro-l H-tetrazol-I-yl) Column:
ethyl)-4-(methoxymethyl)piperidin-4-yl)-N- - size: 1= 0.15 m, 0 = 4.6 nun;
phenylbutanamide. - 'tauonary phase: base-deactivated end-capped octaduylsi1y1
_______________ ~ !'IIE" sllica gelfor chromatography R (5 um);
- temperature: 25°C; if necessary, increase the temperature
slightly to achieve the required resolution between the
peaks due to impurity G and alfuzosin.
Mobile phase Mix I volume of telrahydrofuran R, 20 volumes
Alfuzosin Hydrochloride of acetonitrlle Rand 80 volumes of a solution prepared as
(Ph. Bur. monograph 1287) follows: dilute 5.0 mL of perchloric acidR in 900 mL of water
for chromaUJgraphy R, adjust to pH 3.5 with dilute sodium
CH, hydroxide solution R and dilute to 1000 mL with waterfor
H'C0XX;N"¥,,N~~ H./I chromatography R.
I I Y '0--' , HCI Flow rate 1.5 mUmin.
H ~ ~N 0
3CO Detection Spectrophotometer at 254 run.
NH2 and enentcmer Injedon 10 ~L
Run time Twice the retention time of alfuzosin.
425.9 81403-68-1 Identification of impurities Use the chromatogram supplied
with alfuzosin for sy,tem ,uitabilityA CRS and the
Action and use
Alphaj-adrenoceptcr antagonist.
identify the peaks due to impurities B, F and G; use the
Preparations chromatogram supplied with alfuzosin for peak
Alfuzosin Tablets identification CRS and the chromatogram obtained with
Alfuzosin Prolonged-release Tablets reference solution (c) to identify the peak due to impurity D.
PhE" _ Relative retention With reference to alfuzosin (retention
time ;;;; about 9 min): impurity D ;;;; about 0.4;
DEFINITION = =
impurity B about 0.57; impurity F about 0.63;
(2RSj-N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2-y1) impurity G = about 0.9.
methylamino)propyl]oxolan-2-carboxamide hydrochloride. System suitabIlity Reference solution (b):
Content - resolution: minimum 1.5 between the peaks due to
99.0 per cent to 101.0 per cent (anhydrous substance). impurities Band F; minimum 1.5 between the peaks due
to impurity G and alfuzosin.
CHARACTERS
Appearance Limits:
Whiteor almost white, crystalline powder, slightly - correction factor. for the calculation of content, multiply the
hygroscopic. peak area of impurity F by 0.6;
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1-98 Alginic Acid 2022
- impwlly D: not more than twice the area of the principal

- impun"ty F: not more than 1.5 times the area of the
principal peak in the chromatogram obtainedwith
- unspecified impurities: for each impurity, not more man the D. 1'1'-(3-antinopropyl)-6,7-dimethoxy-N"-methylquinazolin-
area of the principal peak in the chromatogram obtained 2,4-diamine,
in the chromatogram obtainedwith reference solution (a)
(0.3 per cent);
- disregard lim£l: 0.5 times the area of the principal peak in
(0.05 per cent).
Water (2.5.12) E. N-[3-[ (4-amino-6,7-dimethoxyquinazolin-2-yl)
Maximwn 0.5 per cent, determined on 1.00 g. methylamino]propyl]fonnamide,
ASSAY
Dissolve 0.300 g in a mixture of 40 mL of anhydrous acetic
acid Rand 40 mL of acetic anhydride R. 'Titrate with 0.1 M
perchlmU acid, determining the end-point potentiometrically
(2.2.20).
I mL of 0.1 M perch/oric acid is equivalent to 42.59 mg F. 6,7-dimethoxy-N',N"-dimethylquinazoline-2,4-diamine,
of C"H28CIN,O,.
STORAGE
In an airtight container, protected from light.
IMPURITIES
Specified impurities D, F.
Otherdet«table impurities (lhe following substances would, if
present at a sufficient level, be detected by one or otherof the tests G. 1'1'-[3-[(4-antino-6,7-dimethoxyquinazolin-2-yl)aminoj
in the monograph. They are limited by the general aeuptanu propylj-6,7-dimethoxy-N'-methylquinazoline-2,4-diantine.
criterion for other/unspecified impurities andlor by the general _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEor
monograph Substances for phannaceuticol use (2034). It is
therefore not necessary UJ identify these impun'tres for
demonstration of romplianc~ See aha 5.10. Control of impurilies
in substances for pharmaceutical use) A, B, C, E, G.
Alginic Acid
Action and use
Treatment of gastro-oesophageal reflux disease; excipient;
thickening agent.
PhEor ~ _
A. N- [3-[(4-antino-6,7-dimethoxyquinazclin-2-yl)
methylantino]propyl]furan-2-catboxamide, DEFINITION
Mixture of polyuronic acids [(C,HsO,).] composed of
residues of D-mannuronic and L-guluronic acids, obtained
mainly from algae belonging to the Phaeophyceae. A small
proportion of the carboxyl groups may be neutralised.
Content
19.0 per cent to 25.0 per cent of carboxyl groups (-C02 H )
B. 2-cWoro-6,7-dimethoxyquinazolin-4-amine, (dried substance).
CHARACTERS
Appearance
White or pale yellowish-brown, crystalline or amorphous
powder.
Solubility
Very slightly soluble or practically insoluble in ethanol
(96 per cent), practically insoluble in organic solvents.
C. (2RS)-N-[3-[(4-antino-6,7-dimethoxyquinazolin-2-yl) It swells in water but does not dissolve; it dissolves in
amino]propyl]-N-methyloxolan-2-carboxamide, solutions of alkali hydroxides.
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2022 A1imemazine Tartrate 1-99
IDENTIFICATION Particle-size distribution (2.9.31 or 2.9.3/f)

A. To 0.2 g add 20 mL of water Rand 0.5 mL of sodium Settling volume
carbonate solution R. Shakeand filter. To 5 mL of the filtrate Place 75 mL of waler R in a 100 mL graduated cylinder and
add 1 mL of calcium chloride solution R. A voluminous add 1.5 g of the substance to be examined in 0.5 g portions,
gelatinous mass is formed. shaking vigorously after each addition. Dilute to 100.0 mL
B. To 5 mL of the filtrate obtained in identification test A with water R and shake again until the substance is
add 0.5 mL ofa 123 gIL solution of magnesium sulfate R. homogeneously distributed. Allow to stand for 4 h and
No voluminous gelatinous mass is formed. determine the volume of the settled mass.
C. To 5 mg add 5 mL of water R, I mL of a freshly Thefollowing characteristic may be relevant for alginic acid used
prepared 10 gIL solution of 1,3-di/tydro:<yTlaphthakTle R in as geUing agentor viscosity-increasing agent.
ethanol (96 per cellO Rand 5 mL of hydrochloric acid R. Boil Apparent viscosity
gently for 3 min, cool, add 5 mL of water R, and shake with Determine the dynamic viscosity using a rotating viscometer
15 mL of di-isopropyl ether R. Carry out a blank test. (2.2. IIJ).
The upperlayerobtained with the substance to be examined
Prepare a 20 gIL suspension of alginic acid (driedsubstance)
exhibits a deeperbluish-red colour than that obtained with
and add 0.1 J\1 sodium hydroxide until a solution is obtained.
the blank. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE«
TESTS
Chlorides
Maximwn 1.0 per cent.
To 2.50 g add 50 mL of dJute Tlitric acid R, shake for I h Alimemazine Tartrate
and dilute to 100.0 mL with dilute nitric acid R. Filter.
To 50.0 mL of the filtrate add 10.0 mL of 0.1 M silvernitrate (Alimemazine Hemiumraie Ph. Bur. monograph
and 5 mL of toluene R. Titrate with O. J M ammonium 2650)
thiocyanate, using 2 mL of/em', ammonium sulfate solution R2
as indicator and shaking vigorously towards the end-point.
1 mL of 0.1 M silver nitrate is equivalent to 3.545 mg of Cl.
Los. on drying (2.2.32) and erenucmer •
H OH
H0:2CXyC0:2H
1
Maximum 15.0 per cent, determined on 0.1000 g by drying
in an oven at 105 DC for 4 h.
f H OH 'I,

Maximum 8.0 per cent (dried substance), determined on C,oH2,N20,S 373.5 433IJ.99-8
0.100 g.
Microbial contamination Action and use
TAMC: acceptance criterion 102 CFU/g (2.6.12). Histamine Hj, receptor antagonist; sedative.
Absence of Escherichia coli (2.6.13). Preparations
Absence of Salmonella (2.6.13). Paediatric Alirnemazine Oral Solution
StrongPaediatric Alimemazine Oral Solution
ASSAY
To 0.2500 g add 25 mL of waterR, 25.0 mL of O. 1 M Alirnemazine Tablets
sodium hydroxide and 0.2 mL of phenolphthakiTl solution R. PhE« _
Titrate with 0.1 M hydrochlori< acid.
I mL of 0.1 M sodium hydroxide is equivalent to 4.502 mg of DEFINITION
carboxyl groups (-C02lI). (2RS)-N,N,2-Trimethyl-3-( IOH-phenothiazin-I O-yl)propan-
I-amine hemi[(2R,3R)-2,3-dihydroxybutanedioate].
FUNCTIONALITY-RELATED CHARACTERISTICS
Content
This section provides infonnation on charaaeristics that are 99.0 per cent to 101.0 per cent (dried substance).
recognised as being relevant control parameters far oneor more
functions of the substance when usedas an excipient (see chapter CHARACTERS
5.15). Someof the chorocterisucs described in the Functionality- Appearance
related characteristics sutton may also bepresent in the mandatory White or very slightly yellowish powder.
part of the monograph since they alsorepresent mandatoryqua/ily Solubility
criutia. In such cases, a cross-reference UJ the tests described in the Freelysoluble in water, sparingly soluble in ethanol
mandatory pare is induded in the Funaionaiiey-related (96 per cent), practically insoluble in toluene.
characteristics section. Control of the characteristics can contribute It deteriorates when exposed to air and light.
to the quality of a medicinal produa by improoing the consistency
of the manufacturmg process and the perfonnance of the medicinal IDENTIFICATION
product dun'ng use. Where control methods are cited, they are Infrared absorption spectrophotometry (2.2.24).
recognised as beingsuiuzble for the purpose, but othermethods can Comparison alimemasine hemisanrate CRS.
alsobe used. W1Jerever results for a particular characteristic are
TESTS
reported, the control method must be indicated.
Thefollowing characteristics may be relevant for alginic acid used The solution is not more opalescent than reference
as disintegrant and/or binder. suspension II (2.2.1) and not more intensely coloured than
reference solution BY5 (2.2.2, Mehod if).
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1-100 Allantoin 2022
Dissolve 1.0 g in water R and dilute to 10 mL with the same ASSAY

solvent. Dissolve 0.300 g in 50 mL of anhydrous ac.en"c acid R. Titrate
pH (2.2.3) with 0.1 IH perchloric acid, determining the end-point
5.0 to 6.5. Cany out the tesl protected from lightand use a potentiometrically (2.2.20).
freshly prepared solution. I mL of 0.1 M perehloric acid is equivalent to 31.35 mg of
Dissolve 1.0 g in carbon dioxide-free waterR and dilute to C,oH"N,03S,
50 mL with the same solvent. STORAGE
Related substances In an airtight container, protected from light.
Liquid chromatography (2.2.29). Cany au' the test protected IMPURITIES
from light and use freshly prepared solutions.
Specified impurities A, B, C.
Solvent mixture aatonitrile R, waterR (20:80 VIV).
the solvent mixture. and enantiomer

Reference solution (b) Dissolve .3.5 mg of alimemazine for A. (2RS)-N,N,2-trimethyl-3-(5-oxido-l OH-phenothiazin- I0-
system suitability CRS (containing impurities A, B and C) in yl)propan-I-amine,
the solvent mixture and dilute (0 10.0 mL with me solvent
mixture.
Column:
- size: 1= 0.15 m, 0 = 4.6 mm;
- stationary phase: base-deacuVal<d end-capped octadecylsayl
silica gelfor,chromatography R (3 pm);
kfobilephase acetonitrile RJ methanol R, 3.854 gIL solution of B. (2RS)-N,2-dimethyl-3-(IOH-phenothiazin-IO-yl)propan-l-
ammonium acetal< R (10:40:50 VIVIV). amine,
Flow ral< 1.3 mUmin.
~NH
Detection Spectrophotometer at 253 om.
Injedon 20~.
Run time Twice the retention time of alimemazine.
s~
ldentificau·on of impurities Use the chromatogram supplied
with alimemazine for system suitability CRS and the
U
identify the peaks due to impurities A, B and C. C. JOH-phenothiazine.
Relative retention With reference to alimemazine (retention _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POE"
time = about 27 min): impurity A = about 0.1;
=
impurity B about 0.5; impurity C = about 1.4.
- resolution: minimum 5.0 between the peaks due to Allantoin *****
alimemazine and impurity C. ** **
Co/culation of percentage cantents: (ph. Eur. mo"ograph 1288) ***
- correction facto": multiply the peak areas of the following
impurity A = 4.4; impurity C = 0.4; and enanliomer
- for each impurity, use the concentration of alimemazine
hernitartrate in reference solution (a).
Limits:
158.1 97-59-6
- impurities A, C: for each impurity, maximum Action and use
0.15 per cent; Astringent; keratolytic.
0.10 per cent; Pl>E" ~ _
- total: maximum 0.5 per cent; DEFINITION
- reporting threshold: 0.05 per cent. Allantoin contains not less than 98.5 per cent and not more
Loss on drying (2.2.32) than the equivalent of 101.0 per cent of (RS)-(2,5-
Maximum 0.5 per cent, determined on 1.000 g by drying in dioxoimidazolidin-4-yJ)urea.
CHARACTERS
Sulfated ash (2.4.14) A white or almost white, crystalline powder, slightly soluble
Maximwn 0.1 per cent, determined on 1.0 g. in water, very slightly soluble in alcohol.
It melts at about 225°C, with decomposition.
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2022 Allergen Products 1-101
IDENTIFICATION solution of dimethylaminobenzaldehyde R in a mixture of

First identification: A. I volume of hydrochloric acid Rand 3 volumes of methanol R.
Second identification: BJ C, D. Dry the plate in a current of hot air. Examine in daylight
after 30 min. Any spot in the chromatogram obtained with
A. Examine by infrared absorption spectrophotometry
test solution (a), apart from the principal spot, is not more
(2.2.24), comparing with the spectrum obtained with
intense than the spot in the chromatogram obtained with
allantain CRS.
reference solution (b) (0.5 per cent). The test is not valid
B. Examine the chromatograms obtained in the test for unless the chromatogram obtained with reference solution (c)
related substances. The principal spot in the chromatogram shows two clearly separated principal spots.
obtained with test solution (b) is similar in position, colour
and size to the principal spot in the chromatogram obtained Loss on drying (2.2.32)
with reference solution (a). Not more than 0.1 per cent, determined on 1.000 g by
drying in an oven at 105 "C.
C. Boil 20 mg with a mixture of 1 mL of dilute sodium
hydroxide solution Rand 1 mL of waterR. Allow to cool. Sulfated ash (2.4.14)
Add I mL of dilutehydrochloric acid R. To 0.1 mL of the Not more than 0.1 per cent, determined on 1.0 g.
solution add 0.1 mL of a 100 gIL solution of potassium ASSAY
bromide R, 0.1 mL of a 20 gIL solution of resorcinol Rand Dissolve 120.0 mg in 40 mL of water R. Titrate with 0.1 M
3 mL of su/fun'c add R. Heat for 5 min to 10 min on a water- sodium hydroxide, determining the end-point
bath. A dark blue colour develops, which becomes red after potentiometrically (2.2.2fJ).
cooling and pouring into about 10 mL of water R.
1 mL of 0.11H sodium hydroxide is equivalent to 15.81 mg of
D. Heat about 0.5 g. Ammonia vapour is evolved, which C,HoN,03.
turns red litmus paper R blue.
IMPURITIES
TESTS
Solution S
Dissolve 0.5 g in carbon dioxide-free water R, with heating if
necessary, and dilute to 100 mL with the same solvent.
Acidity or alkalinity A. glyoxylic acid,
To 5 mL of solution S add 5 mL of carbon dioxide-free
waterR, 0.1 mL of methylred solution Rand 0.2 mL of
0.01 M sodium hydroxide. The solution is yellow. Add 0.4 mL
of o. 01 M hydrochloric aCUl. The solution is red.
Optical rotation (2.2. 7)
The angle of optical rotation, determined on solution S, is B. carbamide (urea).
-0.10' to + 0.10'. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'"
Reducing substances
Sbake 1.0 g with 10 mL of waterR for 2 min. Filter.
Add 1.5 mL of 0.02 M potassium permanganate. The solution
Allergen Products ***
*** ***
must remain violet for at least 10 min.
Related substances
Examine by thin-layer chromatography (2.2.27), using a
(ph. Eur. monograph 1063) ***
PhE'" _
suitable cellulose for chromatography R as the coating
substance. This monograph does not apply to: chemicals lhal are usedsolely
Testsolution (a)Dissolve 0.10 g of the substance to be for diagnosis of contact dermacuis: chemically synthesised produas;
examined in 5.0 mL of waterR with heating. Allow {O cool. aOergens derived by recombinant DNA lUhnoiogy. It does nor
Dilute to 10 mL with methanol R. Use the solution immediately necessarily apply to allergen products for veten·nary use.
after preparauon.
DEFINITION
Test soludon (b) Dilute I mL of test solution (a) to 10 mL
Allergen products are pharmaceutical preparations derived
with a mixture of 1 volume of methanol R and I volume of
from extracts of naturally occurring source materials
waterR. containing allergens, which are substances that lead to andlor
Reference solution (a) Dissolve 10 mg of al/antoin CRS in a provoke allergic reactions. The allergenic components are
mixture of I volume of methanol Rand 1 volume of water R most often of a proteinaceous nature. Allergen products are
and dilute to 10 mL with the same mixture of solvents. intended for in vivo diagnosis or treatment of allergic diseases
Reference solution (b) Dissolve 10 mg of urea R in 10 mL of attributed to these allergens.
waterR. Dilute 1 mL of this solution {O 10 mL with Allergen products are available as finished products, and as
methanol R. finished products used on a named-patient basis. Allergen
Reference solution (c) Mix I mL of reference solution (a) products are generally presented as parenteral preparations,
and 1 mL of reference solution (b). eye preparations, preparations for inhalation, preparations for
Apply to the plate 10 ~L of test solution (a) and 5 ~L each oral use, sublingual preparations or preparations for skin
of test solution (b), reference solution (a), reference tests.
solution (b) and reference solution (c). Develop over a path For in vivo diagnostic use, allergen products are usually
of 10 em using a mixture of IS volumes of glacial acetic prepared as unmodified extracts in a 50 per cent VIV
acid R, 25 volumes of waterRand 60 volwnes of butanol R. solution of glycerol for skin testing. For intradermal diagnosis
Allow the piate ro dry in air. Spray the plate with a 5 gIL or for provocation tests by nasal, ocular or bronchial
administration, suitable dilutions of allergen products may be
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1-102 Allergen Products 2022
prepared by dilution of aqueous or glycerinated extracts, or pharmaceutical preparations and substances for pharmaceutical
by reconstitution of unmodified freeze-dried extracts. use.
For specific immunotherapy, allergen products may be either All allergen preparations are manufactured under conditions
unmodified extracts or extracts modified chemically and/or designed to minimise exogenous and endogenous enzymatic
by adsorption onto different carriers (for example, aluminium degradation.
hydroxide, calcium phosphate or tyrosine). Any purification procedure is designed to minimise the
PRODUCTION content of any potential irritant low-molecular-mass
Where allergen products or source materials are components and non-allergenic components.
manufactured using materials of human or animal origin, the Allergen products may contain suitable antimicrobial
requirements of general chapter 5.1.7. Viral safety apply. preservatives. The nature and concentration of the
SOURCE MATERIALS antimicrobial preservatives have to be justified and their
Source materials are obtained from qualified suppliers. efficacy complies with chapter 5.1.3. Efficacy of antimicrobial
The source materials comply with the requirements of the preseroouon.
appropriate individual monographs (where a relevant The manufacturing process comprises various stages:
monograph exists) and the statements in this section are - source material;
intended to be read in conjunction with the individual - active substance: it is generally a modified or an
monographs. unmodified allergen extract; where applicable it is stored
Source materials for the preparation of allergen products are under conditions ensuring its stability, for example freeze-
products of animal or vegetable origin, mostly pollens, dried;
moulds, mites, animal epithelia and outgrowths, and - finished product.
Hymenoptera venoms. Other source materials include certain All other stages of the manufacturing process are considered
insects and foods. as intermediates,
The source materials are defined, where possible, by their IN-HOUSE REFERENCE PREPARATION
origin, nature, method of collection or production and pre- An appropriate representative preparation is selected as the
treatment. Control methods and acceptance criteria relating in-house reference preparation (llIRP») characterised and
to identity and purity are established. The acceptance criteria used to verify batch-to-batch consistency. The IHRP is
must ensure the consistency of the allergenic source material stored in suitably sized aliquots under conditions ensuring its
from a qualitative and quantitative point of view. The source stability, for example freeze-dried.
materials are stored under controlled conditions justified by Characterisation of the in-house reference preparation
stability data. . The extent of characterisation of the IHRP depends on the ,ou""
The collection or production, as well as the handling of the material, knowledge of the allergenic components and availabjli~
source materials, are such that consistent composition is of suitable reagents, as well as the intended use. The characrerised
ensured from batch to batch. IHRP is usedas the reference in the batch control of active
When applicable, pesticides, heavy metals and residual substances and imennediates and, if possible, in the batch control
solvents are limited according to the principles defined in of finished products.
general chapters 2.8.13. Pesticide residues, 2.4.27. Heavy metals The IHRP is characterised by the protein content
in herlJai drugs and herbal drugpreparahons and 1.4.24. determination and a protein profile using appropriate
Identification and control of residual solvents, respectively. methods (such as isoelectric focusing, sodium dodecyl sulfate
M.icrobial contamination of the source material may be polyacrylamide gel electrophoresis, immunoelectrophoresis,
unavoidable and should be monitored according to a justified capillary electrophoresis, chromatographic techniques and
sampling plan; if a determination of microbial contamination mass spectrometry).
is not applicable, this must be justified. Allergenic components may be detected by appropria te
The scientific name (species, variety, strain etc.) of the source methods (for example, immunobloning or crossed radio-
material is indicated and the part used is stated, if applicable. immunoelectrophoresis). Characterisation of the allergenic
components may include identification of relevant allergens
Foods must be of a quality suitable for human consumption.
based on serological or other techniques using pooled or
The origin of the food stuff as well as its processing stage is
individual sera from allergic patients, or allergen-specific
stated.
polyclonal or monoclonal antibodies.
MANUFACTURING PROCESS
Determination of the content of relevant allergens is
Allergen products are generally obtained by extraction, and
performed wherever possible. TIlls determination may be
may be purified, from the source materials using appropriate
made using individual allergen-specific reference standards,
methods shown to preserve the allergenic properties of the
when available. The choice of the relevant allergen
components. Allergens for which there are not enough
components subjected to the determination must be justified.
patients to determine the total allergenic activity in vivo or in
Individual allergens are identified and named according to
vitro, the extraction ratio indicating the relative proportions
internationally established nomenclature wherever possible.
(mlV) of allergenic source materials and solvents is a
minimum requirement. Allergen products presented as The biological potency of the first IHRP is determined in
parenteral preparations, eye preparations) preparations for patients by in vivo techniques such as skin testing, and
inhalation and preparations for skin testing are manufactured expressed in units of biological activity, except when not
under aseptic conditions. enough patients are available. In this case, the potency of the
first IHRP is determined by an in vitro method.
In the manufacture, packaging, storage and distribution of
Subsequently, the biological activity of future IHRPs is
allergen products intended for administration by other routes,
demonstrated by in vitro methods by comparison with the
suitable measures are taken to ensure their microbial quality;
results obtained with the first IHRP. The in Wl1'O potency
recommendations on this aspect are provided in general
may be measured by a suitable immunoassay (for example,
chapter 5.1.4. Microbiologi<al quality of non-sterile
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2022 Allopurinol 1-103
an assay based on the inhibition of the binding capacity of determination of individual allergens or arlY otherjustified tests)
specific immunoglobulin E antibodies). must be applied.
IDENTIFICATION Aluminium (2.5. H)
The tests for identification are performed as late as possible 80 per cent to 120 per cent of the stated amount but in any
in the manufacturing process. In the case of products used case not more than 1.25 mg per human dose unless
on a named-patient basis, the control is performed on the otherwise justified and authorised, when aluminium
active substance and/or at the intermediate stage between the hydroxide or aluminium phosphate is used as adsorbent.
active substance and the finished product. Calcium (2.5.1f)
Identity is confirmed by comparison with the IHRP using 80 per cent to 120 per cent of the stated amount when
protein profiling by appropriate methods (for example, calcium phosphate is used as adsorbent.
isoelectric focusing, sodium dodecyl sulfate polyacrylamide Allergen profile
gel electrophoresis, immunoelectrophoresis, immunoblorting, Relevant allergenic components are identified by means of
liquid chromatography or mass spectrometry). suitable techniques using allergen-specific human or animal
In exceptional cases, if no IHRP is available, a representative antibodies.
batch may be used to confirm identity. Total allergenic acdvity
Identity may also be confirmed by comparison with 50 per cent to 150 per cent of the stated amount as assayed
individual allergen-specific reference standards, when by inhibition of the binding capacity of specific
available. immunoglobulin E antibodies or a suitable equivalent in tniro
method.
TESTS
The tests are performed as late as possible in the Individual allergens
manufacturing process. In the case of products used on a 50 per cent to 200 per cent of the stated amount of each
named-patient basis, the control is performed on the active relevant allergen component, determined by a suitable
substance and/or at the Intermediate stage between the active method.
substance and the finished product. STORAGE
Various biochemical and immunological tests have been Adsorbed allergen products are not to be frozen, unless
developed in order (Q characterise allergens qualitatively and otherwise justified and authorised.
quantitatively. In those cases where such methods cannot be
LABELLING
applied, particularly for the determination of allergenic
activity and allergen and/or protein profile, justification must
The labd ,tates:
- the name of the aUergen product;
be provided. .
- the biological potency and/or the protein content and/or
Water (2.5.12 or 2.5.32) or loss on drying (2.2.32) the extraction concentration;
Maximum 5 per cent for freeze-dried products. In the case of - the route of administration and the intended use;
oral Iyophilisates, the water content may be higher than - the storage conditions;
5 per cent, where justified and authorised. - where applicable, the name and amount of added
Sterility (2.6.1) antimicrobial preservative;
Allergen products presented as parenteral preparations, eye - where applicable, for freeze-dried preparations:
preparations, preparations for inhalation or preparations for - the name, composition and volume of the
skin testing comply with. the test for sterility. reconstituting liquid to be added;
Microbial contamination - the period of time within which the preparation is to
For non-sterile allergen products, recommendations are be used after reconstitution;
provided in general chapter 5.1.4. Microbiologi<ol quality of - where applicable, that the preparation is sterile;
non-sunk pharmaceutical preparations and substances for - where applicable, the name and amount of adsorbent.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIf"
pharmaceutical use.
Protein content (2.5.33)
80 per cent to 120 per cent of the stated content, unless
otherwise justified and authorised. If the biological potency
can be determined then the test for protein content is Allopurinol
performed as a batch-to-batch consistency test and the
protein content is within 50 per cent to 150 per cent of the (Ph. Eur. monograph 0576)
stated content. When the finished product contains
proteinaceous excipients, the test for protein content is
o
performed as late as possible during production before
addition of the proteinaceous excipient.
N~NH
.r,
Protein profile ~ N
The protein profile determined by suitable methods

corresponds to that of the IHRP. The presence of relevant 136.1 315-30-0
allergen components is verified, where possible. The choice
of relevant allergen components to be tested for must be Action and use
justified. Xanthine oxidase inhibitor; treatment of gout and
Varia-us additional tests, some with increasing selectivity, hyperuricaemia.
depending on the allergen product concerned can be applied, but in Preparations
airy case for allergen products intended for therapeutic use, a Allopurinol Oral Suspension
validated test measuring the potency (total allergenic activity, Allopurinol Tablets
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1-104 Allopurinol 2022
PhE/¥ _ Test solution (b) Dissolve 20.0 mg of the substance to be

examined in 5.0 mL of a 4 gIL solution of sodium hydroxide R
DEFINITION
and dilute immediately to 250.0 mL with the mobile phase.
1,5-Dihydro-4H-pyrazolo[3,4-djpyrimidin-4-one.
Content 100.0 mL with the mobile phase. Dilute 5.0 rnL of this
CHARACTERS Reference solution (b) Dissolve 5 mg of aUopurinoi
Appearance impurity A CRS, 5 mg of aUopurinoi impurity B CRS and
White or almost whitepowder. 5.0 mg of aUopurinoi impurity C CRS in 5.0 rnL of a 4 WL
Solubility solutionof sodium hydroxide R and dilute immediately to
Very slightly soluble in water and in ethanol (96 per cent). 100.0 rnL with the mobile phase. Dilute 1.0 mL of this
It dissolves in dilute solutions of alkali hydroxides. solutionto 100.0 mL with the mobile phase.
IDENfIFICATION Reference solution (c) Dissolve 20.0 mg of aUopurinoi CRS in
First identification: B. 5.0 rnL of a 4 WL solution of sodium hydroxide R and dilute
immediately to 250.0 mL with the mobile phase.
Column:
A. Ultraviolet and visible absorption spectrophotometry - size: I = 0.25 m, 0 = 4.6 mm;
(2.2.25). - stationary phase: ocrade<y/silyl SIlica gelfor chromatography R
Test solution Dissolve 10 mg in 1 mL of a 4 gIL solution of (5 pm).
sodium hydroxide R and dilute to 100.0 rnL with a 10.3 WL Mobile phase 1.25 WL solution of potassium dihydrogen
solution of hydrochloric <Kid R. Dilute 10.0 rnL of this phosphate R.
solution to 100.0 rnL with a 10.3 WL solution of hydrochltJric
<Kid R.
SpeC/rol range 220-350 run. Delation Spectrophotometer at 230 ron.
Injeuion 20 J.lL of test solution (a) and reference
Absorption maximum At 250 nm.
solutions (a) and (b).
Absorption minimum At 231 nrn.
Run time Twice the retention time of allopurinol.
Absorbance ratio A231/A250 = 0.52 to 0.62.
Elution order Impurity A, impurity B, impurity C,
B. Infrared absorption spectrophotometry (2.2.24). allopurinol.
Compotison allopurinol CRS. Retention time Allopurinol = about 10 min.
C. Dissolve 0.3 g in 2.5 rnL of dilute sodium hydroxide System suitability Reference solution (b):
solution R and add 50 rnL of water R. Add slowly and with - resolution: minimum 1.1 between the peaks due to
shaking 5 mL of silver nitrate solution Rl. A white precipitate impurities Band C.
is formed which does not dissolve on the addition of 5 mL of
ammonia R. Limits:
- impurity A: not more than twice the area of the principal
D. Thin-layer chromatography (2.2.27). peak in the chromatogram obtained with reference
Test solution Dissolve 20 mg of the substance to be solution (a) (0.2 per cent);
examined in concentrated ammonia R and dilute to 10 mL - impurity B: not more than the area of the principal peak in
with the same solvent. the chromatogram obtained withreference solution (a)
Reference solution Dissolve 20 mg of alltJpurinoi CRS in (0.1 per cent);
concentrated ammonia R and dilute to 10 mL with the same - ~"mpurity C: not more than the area of the corresponding
solvent. peak in the chromatogram obtained with reference
Plate TLG silica gel F254 plateR. solution (b) (0.1 per cent);
Mobile phase anhydrous ethanol R, methylene chloride R - unspecified ~'mprmiies: for each impurity) not more than the
(40:60 VIV).
area of the principal peakin the chromatogram obtained
Application I0 ~L. - sum of impurities other thanA, Band C: not more than
Development Over 213 of the plate. 3 times the area of the principal peak in the
Drying In air. chromatogram obtained with reference solution (a)
Detection Examine in ultraviolet lightat 254 nm. (0.3 per cent);
(0.05 per cent).
principal spot in the chromatogram obtained with the
reference solution. Impurities D and E
Liquid chromatography (2.2.29). Usejreshly prepared soludons.
TESTS Store and injut them at 8 °C, "using a cooled aUlOsampkr.
Related substances
Liquid chromatography (2.2.29). Use freshly prepared solmions. Sduuon A 1.25 WL solution of potassium dihydrogen
phosphate R.
Store and inject them at 8 °C, using a cooled autosampler.
Testsolution (a) Dissolve 25.0 mg of the substance to be
examined in 2.5 rnL of a 4 WL solution of sodium hydroxide R examined in 5.0 mL of a 4 WL solution of sodium hydroxide R
and dilute immediately to 50.0 rnL with the mobile phase. and dilute immediately to 100.0 rnL with solution A.
Reference solution Dissolve 5.0 mg of aiiopurinol
impurity D CRS and 5.0 rng of alWpurinoi impurity E CRS in
5.0 rnL of a 4 WL solution of sodium hydroxide R and dilute
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2022 Allopurinol 1-105
immediately [0 100.0 mL with solution A. Dilute 1.0 mL of System suitability Reference solution:
this solution to 100.0 mL with solution A. - resolution: minimum 2 between the peaks due to
Column: benzaldehyde azine and benzaldehyde;
- size: / = 0.05 m, 0 ;;:; 4.6 mm; - signal-to-noise ratio: minimum 20 for the peak due to
- stationary phase: base-deactivated octadecy/silyl silica geljQT benzaldehyde azine.
chromatography R (3 pm). Limit:
Mobile phase methanol R, 1.25 gIL solution of potassium - impurity F: the area of the peak due to benzaldehyde azine
dihydrogen phosphate R (10:90 VIV). in the chromatogram obtained with the test solution is not
more than the area of the corresponding peak in the
Flow rate 2 mUmin.
chromatogram obtained with the reference solution
Detection Spectrophotometer at 230 run. (10 ppm of hydrazine sulfate equivalent to 2.5 ppm of
Injection 20 ~L. hydrazine).
Run time 1.5 times the retention time of impurity E. Loss on drying (2.2.32)
Retention times Impurity D = about 3.6 min; Maximum 0.5 per cent, determined on 1.000 g by drying in
impurity E ;;;;: about 4.5 min. an oven at 105 "C.
System suitability Reference solution: Sulfated ash (2.4.14)
- resolution: minimum 2.0 between the peaks due to Maximum O. I per cent, determined on 1.0 g.
impurities D and E.
ASSAY
Limits: Liquid chromatography (2.2.29) as described in the test for
- impunry D: not more than the area of the corresponding related substances with the following modification.
peak in me chromatogram obtained with the reference
solution (0.1 per cent); Injection Test solution (b) and reference solution (c).
- impurity E: not more than the area of the corresponding Calculate the percentage content of C SH4N"O from the
peak in the chromatogram obtained with the reference declared content of allopurinol CRS.
solution (0.1 per cent). IMPURITlliS
Impurity F Specified impurities A, B, C, DJ EJ F.
Under the following conditions, any hydrazine in the sample
reacts with benzaldehyde to give benzaldehyde azine.
Solvent mixture Mix equal volumes of dilute sodium hydroxide
solution R and methanol R.
Solution A Dissolve 2.0 g of benzaldehyde R in the solvent
mixture and dilute to 50.0 mL with the solvent mixture. A. 5-amino-lH-pyrazole-4-carboxamideJ
Prepare immediately before use.
Test solution Dissolve 250.0 mg of the substance (0 be
examined in 5 mL of the solvent mixture. Add 4 mL of
solution A, mix and allow to stand for 2.5 h at room
temperature. Add 5.0 mL of hexaneR and shake for I min.
Allow the layers to separate and use the upper layer.
Reference solutWn Dissolve 10.0 mg of hydrazine sulfate R in
B. 5-(fonnylamino)-IH-pyrazole-4-earboxamide,
the solvent mixture by sonicating for about 2 min and dilute
to 50.0 mL with the solvent mixture. Dilute 1.0 mL to
solution £0 20.0 mL with the solvent mixture. To 5.0 mL of
the solution obtained, add 4 mL of solution A, mix and
allow to stand for 2.5 h at room temperature. Add 5.0 mL of
hexane R and shake for 1 min. Allow the layers to separate
and use the upper layer.
Blank solution To 5 mL of the solvent mixture add 4 mL of C. 5-( 4H-l ,2,4-triazol-4-yl)-1 H-pyrnzole-4-catboxamide,
solution A, mix and allow to stand for 2.5 h at room
temperature. Add 5.0 mL of hexane R and shake for 1 min.
Allow the layers to separate and use the upper layer.
Column:
- size: I =. 0.25 m, 0 = 4.0 mm;
- suuionory phase: cyanosilyl silica gd for chromatography R
(5 pm) with a pore size of 10 nm;
D. ethyl 5-amino-IH-pyrazole-4-catboxylate,
Mobile phase 2-propanol R, hexane R (5:95 VIV).
Injection 20 1'1-.
Relative retention With reference to benzaldehyde (retention
time = about 2.8 min): benzaldehyde azine = about 0.8.
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1-106 Almagate 2022
Chlorides (2.4.4)
Maximum OJ percent.
Dissolve 0.33 g in 5 rnL of dilute nitric add R and dilute to
100 rnL with water R. Prepare simultaneously the standard
by diluting 0.7 mL of dilute nitric acid R to 5 mL with
E. ethyl 5-(fonnylamino)-IH-pyrazole-4-carooxylate,
waw R and adding 10 mL of chloride standard solution (5 ppm
CDR.
Sulfates (2.4.11)
F. diazane (hydrazine). Dissolve 0.25 gin 5 mL of dilute hydrochlon'c acid Rand
____________________ ""E" dilute to 100 mL with distilled waw R. Prepare
simultaneously the standard by adding 0.8 mL of dilul<
hydrochloric acid R to 15 mL of sulfate standard solution
(10 ppm SO~ R.
Sodium
Almagate Maximum 150 ppm.
(Ph. Eur. monograph 2010) Atontic absorption spectrometry (2.2.23, Method 1).
66827-12-1 Tes' solution Dissolve 0.25 g in 50 mL of a 103 gIL solution
Al,Mll6C,O,oH",4H,O 630
of hydrochloric acidR.
Action and use Reference solutions Prepare the reference solutions using
Antacid. sodium standard solution (200 ppm Na) R, diluted as necessary
""E,, _ with a 103 gIL solution of hydrochloric acid R.
Loss on ignition
DEFINITION 43.0 per cent to 49.0 per cent, determined on 1.000 g by
Hydrated aluminium magnesium hydroxycarbonate. ignition at 900 ± 50 "C.
Content Microbial contamination
- aluminium: 15.0 per cent to 17.0 per cent (calculated TAMC: acceptance criterion 10' CFUlg (2.6.12).
as AJ 2 0 J) , TYMC: acceptance criterion 10' CFU/g (2.6.12).
- magnesium: 36.0 per cent to 40.0 per cent (calculated
as MgO),
- carbonic acid: 12.5 per cent to 14.5 per cent (calculated Absence of Pseudomonas aeruginosa (2.6.13).
as CO,). ASSAY
CHARACTERS Aluminium
Appearance Dissolve 1.000 g in 5 mL of hydrochloric acidR, heating if
White or almost white) fine, crystalline powder. necessary. Allow to cool to room temperature and dilute to
100.0 mL with waw R (solution A). Introduce 10.0 mL of
Solubility
solution A into a 250 mL conical flask, add 25.0 mL of
Practically insoluble in water, in ethanol (96 per cent) and in
0.05 M sodium edetate, 20 mL of buffer solution pH 3.5 R,
methylene chloride. It dissolves with effervescence and
40 mL of anhydrous ethanol Rand 2 mL of a freshly prepared
heating in dilute mineral acids.
0.25 gIL solution of dithizone R in anhydrous ethanol R.
IDENTIFICATION Titrate the excess of sodium edetate with 0.05 M zinc sulfate
A. Infrared absorption spectrophotometry (2.2.24). until the colourchanges from greenish-violet to pink.
Comparison Ph. Eur. reference spectrum of aimagate. 1 mL of 0.05 M sodium edetal< is equivalent to 2.549 mg
B. Dissolve 0.15 g in dilul< hydrochloric acid R and dilute to of Al,O,.
20 mL with the same acid. 2 mL of the solutiongives the Magnesium
reaction of aluminium (2.3. I). Introduce 10.0 mL of solution A prepared in the assay of
C. 2 mL of the solution prepared under identification test B aluminium into' a 500 mL conical flask, add 200 mL of
gives the reaction of magnesium (2.3.1). water R, 20 mL of u;ethanolamine R with shaking, 10 mL of
ammonium chloride buffer solution pH 10.0 Rand 50 mg of
TESTS
mordant black l l triturate R. Titrate with 0.05 M sodium
pH (2.2.3)
edeuue until the colourchanges from violet to pure blue.
9.1 to 9.7.
I mL of 0.05 M sodium ea."'te is equivalent to 2.015 mg
Disperse 4.0 g in 100 mL of carbon dioxide-free waW R, stir
of MgO.
for 2 min and filter.
Carbonic acid
Neutrallsing capacity 12.5 per cent to 14.5 per cent.
Cany ou, the I<st at 37"C Disperse 0.5 g in 100 mL of
waw R, heat, add 100.0 mL of 0.1 M hydrochloric acid, Test sample Place 7.00 mg of the substance to be examined
previously heated and stir continuously; the pH (2.2.3) of the
in a tin capsule. Seal the capsule.
solution between 5 min and 20 min is not less than 3.0 and Reference sample Place 7.00 mg of almagase CRS in a tin
not greater than 4.5. Add 10.0 mL of 0.5 M hydrochloric acid, capsule. Seal the capsule.
previously heated, stir continuously for 1 h and titrate with Introduce separately the test sampleand the reference sample
0.1 M sodium hydroxide to pH 3.5; not more than 20.0 mL of into a combustion chamber of a eHN analyser purged with
0.1 1"1 sodium hydroxide is required. helium for chromatography R and maintained at a temperature
of 1020 "C. Simultaneously, introduce oxygen R at a pressure
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2022 Almond Oil 1-107
of 40 kPa and a flow rate of 20 mUmin and aUow complete Second identification: A, B.
combustion of the sample. Sweep the combustion gases A. Absorbance (see Tests).
through a reduction reactor and separate the gases formed by
B. Identification of fatty oils by thin-layer chromatography
gas chromatography (2.1.28).
(2.3.2).
Column:
~tSUllS The chromatogram obtained is similar to the
- size: 1= 2 m, 0::::: 4 mm;
corresponding chromatogram shown in Figure 2.3.2.-1.
- stationary phase: echy/vjnylhenzene-divinylbenzene
",polymer R. C. Composition of fatty acids (see Tests).
Cam....gas helium for chromatography R. TESTS
Flow rate 100 mUmin. Specific absorhance (2.2.25)
Maximum 0.2, determined at the absorption maximum at
Temperature:
270 om. The ratio of the absorbance measured at 232 om to
- column: 65°C;
that measured at 270 om is greater than 7.
- detector; 190°C.
To 0.100 g add cydohexane R and dilute to 10.0 mL with the
same solvent. Adapt the concentration of the solution so that
Run time 16 min. the absorbance lies between 0.5 and 1.5, measured in a I em
System suitability: cell.
- average percentage of carbon in 5 reference samples must Acid value (2.5.1)
be within ± 0.2 per cent of the value assigned to Maximum 2.0, determined on 5.0 g.
the CRS; the difference between the upper and the lower
values of the percentage of carbon in these samples must Peroxide value (2.5.5, Method A)
he below 0.2 per cent. Maximum 15.0.
Calculate the percentage content of carbonic acid in the test Unsaponifiable mailer (2.5.7)
sample according to the following formula: Maximum 0.9 per cent) determined on 5.0 g.
Composition of fatty acids (2.4.22, Method A)
A
c-ac-.-:
m
Use the mixture of calibrating substances in Table 2.4.22.-3.
Composition of thefatty-add fraction of the oil:
C percentage cement of carbonic acid in the reference sample; - saturated/any adds of chain length less than C 16 : maximum
K mean value for the 5 reference samples of the ratio of the mass 0.1 per cent)
in miUignuns to the area of the peak due [0 carbonic acid; - palmitic acid: 4.0 per cent to 9.0 per cent)
A area of the peak due to carbonic acid in the chromatogram - palm;lQleic acid: maximum 0.8 per cent)
obtained with the test sample;
m sample mass, in milligrams.
- margosic add: maximum 0.2 per cent)
-. stearic acid: maximum 3.0 per cent)
- oleic acid: 62.0 per cent to 86.0 per cent,
STORAGE
- linoleic add: 20.0 per cent to 30.0 per cent)
In an airtight container. - linolenic acid: maximum 0.4 per cent)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
- arachidic acid: maximum 0.2 per cent,
- eicosenoic add: maximum 0.3 per cent)
- behenic add: maximum 0.2 per cent)
- erucic add: maximum 0.1 per cent.
Virgin Almond Oil Sterols (2.4.23)
Composition of sterol fraction of the oil:
Almond Oil - cholesterol: maximum 0.7 per cent)
(Ph. Bur. monograph 0261) - campeuerd: maximum 4.0 per cent)
Preparation - stigmasterol: maximum 3.0 per cent,
Almond Oil Ear Drops - P-sitosterol: 73.0 per cent to 87.0 per cent)
PhE" _ - LJ5-avenasterol: minimum 10.0 per cent)
- Ll7-sugmastenol: maximum 3.0 per cent)
DEFINITION - LJ 7-avenasterol: maximum 3.0 per cent,
Fatty oil obtained by cold expression from the ripe seeds of - brassicasterol: maximum 0.3 per cent.
Pmnus dukis (Mill.) D.A.Webb var. dulcis or Pnmus dulcis Water (2.5.32)
(Mill.) D.A.Webb var. amara (DC.) Buchheim or a mixture Maximum 0.1 per cent, determined on 1.00 g.
of both varieties.
STORAGE
CHARACfERS
In a weU-fiUed container, protected from light.
Appearance _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Yellow, clear liquid.
Solubility
Slightly soluble in ethanol (96 per cent), miscible with light
petroleum.
Relative density
Abour 0.916.
It solidifies at about-IS "C.
IDENTIFICATION
First identification: A, C.
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1-108 Almond Oil 2022
- A5-avenasterol: minimum 5.0 per cent;

Refined Almond Oil - A7-stigmascenol: maximwn 3.0 per cent:
(Ph. Eur. monograph 1064) - A7-avenasterol: maximum 3.0 per cent;
- brassicasterd: maximum 0.3 per cent.
PhEtr _
Water (2.5.31)
DEFINITION Maximum 0.1 per cent, determined on 1.00 g.
Fatty oil obtained from the ripe seeds of Prnnus dulcis STORAGE
(MiU.) D.A. Webb var. duleis or Prunus dukis (Mill.) D.A.
In a well-filled container, protected from light.
Webb var. amara (DC.) Buchheimor a mixture of both
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
varieties by cold expression. It is then refined. A suitable
antioxidant may he added.
CHARACTERS
Appearance
Pale yellow, clear liquid.
Almotriptan Malate
Solubility (Ph Eur. monograph 2970)
Slightly soluble in ethanol (96 per cent), miscible with light
petroleum.
Relative density
About 0.916.
It solidifies at about -18 "C.
IDENTIFICATION andenanUomer
A. Identification of fatty oils by thin-layer chromatography
(2.3.2).
469.6 181183-52-8
Results The chromatogram obtained is similar to me
corresponding chromatogram shown in Figure 2.3.2.-1. Action and use
B. Composition of fatty acids (see Tests). Serotonin 5HTJ receptor agonist; treatment of migraine.
TESTS PhE" _
Speclfic absorbance (2.2-.25)
0.2 to 6.0, determined at the absorption maximum at DEFINITION
270 om. N,N-Dimethyl-2-[5-[(pyrrolidine-l-sulfonyl)methyl]-IH-
indol-3-yl]ethan-l-amine (RS)-2-hydroxybutanedioate.
To 0.100 g add cyclohexane R and dilute to 10.0 mL with the
same solvent. Adaptthe concentration of the solutionso that Content
the absorbance lies between 0.5 and 1.5, measured in a 1 em 98.0 per cent to 102.0 per cent (dried substance).
cell. CHARACTERS
Acid value (2.5.1) Appearance
Maximum 0.5, determined on 5.0 g. White or slightly yellow, crystalline powder.
Peroxide value (2.5.5, Method A) Solubility
Maximum 5.0. Soluble in water, slightly soluble in methanol, practically
Unsaponlfiable matter (2.5.7) insoluble in anhydrous ethanol and in heptane.
Maximum 0.9 per cent, determined on 5.0 g. IDENTIFICATION
Composition of fatty acids (2.4.22, Method A) A. Infrared absorption spectrophotometry (2.2.24).
Use the mixture of calibrating substances in Table 2.4.22.-3. Comparison almotriptan malate CRS.
Composition of thefrmy-acid fraction of the oil: B. Examinethe chromatograms obtained in the assay.
- saturated fatty acids of chain length less than C 16 : maximum Results The principal peak in the chromatogram obtained
0.1 per cent; with test solution (b) is similar in retention time and size to
- palmitic acid: 4.0 per cent to 9.0 per cent; the principal peak in the chromatogram obtained with
- palmitoleic acid: maximum 0.8 per cent; reference solution (b).
- margatic add: maximum 0.2 per cent;
- steanc acid: maximum 3.0 per cent; TESTS
- oleic acid: 62.0 percent to 86.0 per cent; Related substances
- linoleic acid: 20.0 per cent 10 30.0 per cent; Liquid chromatography (2.2.29). Carry out ,he teu prouaed
- linolenk acid: maximum 0.4 per cent; from ligh,.
- arachidic acid: maximum 0.2 per cent; Solvent mixture acetonitrile RJ waterR (20:80 VIV).
- eicosenoic acid: maximum 0.3 per cent; Buffer solution Dissolve 0.5 g of sodium octanesulfonare
- behenic acid: maximum 0.2 per cent; monohydrate R in about 900 mL of water for
- erucic acid: maximum 0.1 per cent. chromatography R. Add 5 mL of phosphonc acid R, adjust to
Sterols (2.4.23) pH 3.0 with sodium hydroxide solution R and dilute to
Compositi<>n of the sterol fraction of the oil: 1000 mL with waterfor chromatography R.
- cholesterol: maximum 0.7 per cent; Test solution (a) Dissolve 10.0 mg of the substance to be
- campeuerd: maximum 5.0 per cent; examined in the solvent mixture and dilute to 10.0 mL with
- stigmasterol: maximum 4.0 per cent; the solventmixture.
- fJ-sitosterol: 73.0 per cent to 87.0 per cent;
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2022 Almotriptan Malate 1-109
Test solution (b) Dissolve 25.0 mg of the substance to be Mobile phase acetonitrile for chromatography R, buffer solution
examined in the solvent mixture and dilute to 50.0 mL with (27:73 VII').
the solvent mixture. Dilute 5.0 mL of the solution to Injection 10 J..IL of test solution (b) and reference
50.0 mL with the solvent mixture. solution (b).
Reference solution (a) Dilute 1.0 mL of test solution (a) to Run time Twice the retention time of almotriptan.
Retention time Almotriptan = about 9 min.
Calculate the percentage content OfC21H3lN307S taking
Reference solutwn (b) Dissolve 25.0 mg of almotriptan into account the assigned content of almompmnmalateCRS,
malate CRS in the solvent mixture and dilute to 50.0 mL
with the solvent mixture. Dilute 5.0 mL of the solution to IMPURITffiS
50.0 mL with the solvent mixture. Speajied impurities A.
Reference solution (c) Dissolve 5 mg of a/motriptan for system Otherdet«tab/e impun·ties (thefollowing subsronces would) if
su;robj/ity CRS (containing impurity A) in the solvent mixture present al a sufficienl/eve!, bedetected by oneor ocher of the tests
and dilute to 5 mL with the solvent mixture. in the monograph. They are limited by the general a«eptarn:e
Column: criterion for otherlunspedfied impurities and/or by thegeneral
- size: 1= 0.25 m, 0 == 4.6 mm; monograph Substances for pharmaceutical use (2034). It is
- stationary phase: end-tapped ottylsilyl silica gelfor therefore not necessary to identify these impurities for
chromawgraphy R (5 urn). demonstration of aJmpliance. See also 5.10. Control of impurities
in subsumces for pharmaceutical use) B) C, D, E, F.
MoMe phase:
- mobile phase A: acewnitrile for chromawgraphy R, buffer
solution (10:90 VII');
- mobile phase B: buffer solution, aaronitn"le for
chromawgraphy R (30:70 VII');
Time MobUe phose A MobUe phase B

(min) (per cent V/J? (per cent V/J?
0-5 85 15
5 - 20 85 ~ 78 15 ..... 22 A. N-methyl-2-[5-[(Pyrrolidine-I-sulfonyl)methyl]-IH-indol-
20 - 30 78 ~ 70 22 ..... 30 3-yl]ethan-I-amine,
30 - 40 70 30
F/()W rOle 1,0 mllmin.

Daeaion Spectrophotometer at 228 nm.
o\\ 0
Injection 5
and (c).
J.1l. of test solution (a) and reference solutions (a)
Identification of impurities Use me chromatogram supplied

O,S II
with almotriptan for system suitability CRS and the

the peak due to impurity A.
B. 2-[2-[[3-[2-(dimethylamino)ethyl)-IH-indol-5-yl]methyl]-
Relative retention With reference to almorriptan (retention
5-[(pyrrolidine-I-suifonyl)me thyl]-I H-indol-3-yl]-N,N-
time = about 27 min): malic acid = about 0.1;
dimeiliylethan-I-amine,
impurity A = about 0.96.
Systemsuitability Reference solution (c):
impurity A and almotriptan.
Colcutasion of percentage contents:
- for each impurity, use the concentration of almotriptan
malate in reference solution (a),
Limits:
- impun·cy A: maximum 0.5 per cent;
- unspecified impun'ties: for each impurity, maximum C. [3-[2-(dimethylamino)ethyl]-5-[(pyrrolidine-l-sulfonyl)
0.10 per cent; metbyl]-IH-indol-l-yl] methanol,
- reporting threshold: 0.05 per cent; disregard the peak due to
malic acid.
D. 2- [5-[(pyrrolidine-I-sulfonyl)methyl]-IH-indol-3-yl]ethan-
Maximum 0.1 per cent. determined on 1.0 g.
l-arnine,
ASSAY
liquid chromatography (2.2.29) as described in the test for
related substances with the following modifications.
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1-11 0 Alprazolam 2022

Preparation Discs.
Comparison alprazolam CRS.
If the spectra obtained in the solid state show dilIerences,
substance separately in the minimum volume of ethyl
E. N ,N-dimethyl-2-[5-[(pyrrolidine-l-sulfonyl)methyl]-IH- aatate R, evaporate to dryness on a water-bath and record
indol-3-yl)ethan-I-amine N-oxide, new spectra using the residues.
examined in methanol R and dilute to 10 mL with the same
solvent.
Reference ,00ution (a) Dissolve 10 mg of alprazolam CRS in
methanol R and dilute to 10 mL with the same solvent.
Reference ,00U1ion (b) Dissolve 10 mg of alprazolam CRS and
F. N-me thyl- N- [2-15-[(pyrrolidine-I-sulfonyl)methyl]-IH- 10 mg of midazolam GRS in methanol R and dilute to 10 mL
. indol-3-yl]ethyl]propan-2-amine. with the same solvent.
__________ ~ PhE" Plate TLC silica gel GFZ54 plate R.
lvlobile phase glacial lUetic acid R, waterR, methanol R, ethyl
acetate R (2:15:20:80 VIVIVIV).
Application 5 ~L.
Alprazolam ***
*** ***
Deudopmen: Over a path of 12 em.
Drying In air.
(Ph. Eur. monograph 1065) ***
- the chromatogram shows 2 clearly separately Spots.
wil:h the test solution is similar in position and size to the
principal spot in the chromatogram obtained with reference
solution (a).
TESTS
Related substances
308.8 28981-97-7 Liquid chromatography (2.2.29).
Buffersolution Dissolve 7.7 g of ammonium acetate R in
Acdon and use 1000 mL of woterfor chromawgraphy Rand adjust to pH 4.2
Benzodiazepine. with glacial acetic acidR.
PhE<I _ Testsolution Dissolve 0.100 g of the substance to be
examined in dimethylfarmamide R and dilute to 10.0 mL with
DEFlNITION the same solvent.
8-Chloro-l-methyl-6-phenyl-4H-[1,2,4] triazoloI4,3-a) Reference ,00ution (a) Dissolve 2 mg of alprazolam CRS and
[1,4)benzodiazepine. 2 mg of triazolam CRS in dimethylfOlmamide R and dilute to
Content 100 mL with the same solvent.
99.0 per cent to 101.0 per cent (dried substance). Reference sohuion (b) Dilute 5.0 mL of the test solution to
CHARACTERS 100.0 mL with dimethylformamide R. Dilute 0.5 mL of this
Appearance solution to 10.9 mL with dimethylfarmamide R.
White or almost white, crystalline powder. Column:
Solubility - size: 1= 0.25 m, 0 = 4.6 rom;
Practicallyinsoluble in water, freely soluble in methylene - 'tationary phase: end-capped extra-dense bonded phenylsi1yl
chloride, sparingly soluble in acetone and in ethanol silica gelfor chromawgraphy R (5 urn).
(96 per cent). Mobile phase:
It shows polymorphism (5.9). - mobile phase A: buller solution, methanol R (44:56 VIV);
- mobile phase B: buller solution, mahand R (5:95 VIV);
IDENTIFICATION - temperature: 40 "C;
Second identification: A, C. Time Mobile phase A Mobile phase B
(min) (per cent VIP) (per cent VIP) .
A. Dissolve the substance to be examined in the smallest
necessary quanrity of ethylacetate R and evaporate to dryness 0-15 98 2
on a water-bath. Thoroughly mix 5.0 mg of the substance to 15 - 35 98 --01 2 --099
be examined with 5.0 mg of alprazolam CRS. The melting 35·40 I 99

point (2.2.14) of the mixture does not differ by more than
2 °C from the melting point of the substance to be Flow rate 2 mIlmin.
examined.
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2022 Alprazolam r-i n

Injection 10 IlL; inject dimethylformamide R as a blank.
Retention time Triazolam = about 9 min;
alprazolam = about 10 min.
System suitability Reference solution (a):
triazolam and alprazolam.
Limits:
D. 8-cbloro-l-ethenyl-6-phenyl-4H-[1,2,4Jlriazolo[4,3-aJ
- total: not more than the area of the principal peak in the
[l}4]benzodiazepine}
~
(0.25 per cent); NH'

'" 1 0
me chromatogram obtained with referencesolution (b) CI
(0.05 per cent).
Loss on drying (2.2.32) I'"
#
an oven at 105 "C.
E. (2-amino-5~b1orophenyl)phenylmethanone,
Sulfuted ash (2.4.14)
ASSAY
Dissolve 0.140 g in 50 mL of a mixture of 2 volumes of
acetic anhydn'de Rand 3 volumes of anhydrous auric acid R.
Titrate with 0.1 M perchlotic acid, determining the end-point
potentiomelrically (2.2.20). Titrate to the 2 nd point of
inflexion.
of C 17H13CIN,. F. [5-cWoro-2-[3-(chloromerhyl)-5-methyl-4H- J,2, 4-lriazol-
4-yl]pbenyl)phenylmethanone,
STORAGE
Protected from light. H,C
~
IMPURITIES
r~N
CI '" 1 """ NH,
and enanllomer
I'"
"""
G. 7~b1oro-l-methyl-5-phenyl[I,2,4Jlriazolo[4,3-a]quinolin
A. (4RS)-3-amino-6-cWoro-2-methyl-4-phenyl-3,4- 4-amine,
dlhydroquinazolin-a-ol,
0 0
N-N N-N
H,C--{ 1\ ~ ">-CH,
N~ ~C(DN
cr¢ J"'-
'" 1
cr
""'"
I ""
CI
""
I
H. bis[[4-(2-benzoyl-4-cWorophenyl)-5-methyl-4H-I,2,4-
lriazol-3-yIJmethyl]amine,
B. [5-cWo';'-2-[3- (hydroxymethyl)-5-methyl-4H-1,2,4-lriazol- H,C"""N

IV ,
~
'" I
4-yl]phenylJphenylmethanone,
N)N N-N
H,C
)=N, CI '. N~ J\.... and enanliomer
~
N -..J' N I' OH N CH3
CI
'" I 0 ~ o~ h
r;7u' ~
CI '"
1#
1. [5-cWoro-2-[3-[[(6RS)-8-cbloro-6-hydroxy-l-methyl-6-
. phenyl-4H-[I,2,4]lriazolo[4,3-aJ [I,4Jbenzodiazepin-5
C. [5-cWoro-2-[3-methyl-4H-l,2,4-lriazol-4-ylJphenyl]
(6H) -yl]methyl) -5-methyl-4H-1,2,4-lriazol-4-ylJpbenylJ
phenylmethanone,
phenylmethanone,
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1-112 Alprenolol Hydrochloride 2022
than reference solution Bg (2.2.2, Method 11).
To 10 mL of solution S add 0.2 mL of methyl red solution R
CI and 0.2 mL of 0.01 M hydrochlOlic acid; the solution is red.
Add 0.4 mL of 0.01 M sodium hydroxide; the solution is
yellow.
J. 2,17-dichloro-6,13-dimethyl-18b,19a-<iiphenyl-
Impurity C
8b,19adihydro-IOH,18bH-[1,2,4]triazolo
[4/ 1',3'":1",2"[quinolo [31f,4" :4',5']oxazolo[3 f ,2' -d]-
1,2,4-triawlo [4,3-0) [1,4]benzodiazepine. Dissolve 0.25 g in ethanol (96 percent) R and dilute to
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf<l
25 mL with the same solvent. The absorbance (2.2.25)
measured at 297 om is not greater than 0.20.
Impurity D
Thin-layer chromatography (2.2.27).
Alprenolol Hydrochloride Test solution (a) Dissolve 0.50 g of the substance [0 be
(Ph. Eur. monograph 0876) solvent.
Testsolution (b) Dilute I mL of test solution (a) to 50 mL
H OH with methanol R.
oe::
~~
?" I yCH,. Hel and enaoucmer Reference solution (a) Dissolve 10 mg of alprenolol
CH, hydrochloride CRS in methanol R and dilute to 10 mL with the
CH,
same solvent.
Reference solutWn (b) Dissolve 10 mg of alprenolol
285.8 13707-88-5 hydrochloride CRS and 10 mg of oxprenolol hydrochloride CRS
in methanol R and dilute to 10 mL with the same solvent.
Action and use
Bera-adrenoceptor antagonist. Reference solucion (c) Dilute 5 mL of test solution (b) to
Phf<l _
Plate TLC silica gel G place R.
DEFINITION Mobile phase Place 2 beakers each containing 30 mL of
(2RS) -1- [( I-Methylethyl)amino)-3- [2-(Prop-2-enyl) ammonia R at the bottomof the tank containing a mixture of
phenoxyjpropan-g-ol hydrochloride. 5 volumes of methanol Rand 95 volumes of ethyl <UetaU R.
Content ApplicatWn 5 1'1-.
99.0 per cent to 101.0 per cent (dried SUbstance). Development Over a path of 15 em in a tank saturated for at
CHARACTERS least 1 h.
Appearance Drying At 100 "C for 15 min.
White or almost white, crystalline powder or colourless Detection Expose to iodinevapour forup to 6 h.
crystals. Syuem suitability Reference solution (b):
Solubility - the chromatogram shows 2 clearly separated spots.
Very soluble in water, freely soluble in ethanol (96 percent) Limits Test solution (a):
and in methylene chloride. - impurity D: any spot with an RF value greater than thatof
IDENTIFICATION the principal spot is not more intense than the principal
First identification: B, D. spot in the chromatogram obtained with reference
solution (c) (0.2 per cent).
Related substances
A. Melting point (2.2.14): 108 "C to 112 "C.
Comparison alprenolol hydrochlonik CRS. examined in the mobilephase and dilute to 10.0 mL with
C. Examine the chromatograms obtained in the test for the mobile phase.
impurity D. Reference sduuon (a) Dissolve 4.0 mg of alprenolol
Detection Examine in daylight, after exposure to iodine hydrochloride CRS and 0.8 mg of 4-isopropj'Iphenal R in the
vapour for 30 min. mobile phase and dilute to 100.0 mL with the mobile phase.
Results The principal spot in the chromatogram obtained Reference soluciotl (b) Dilute 4.0 mL of the test solution to
with test solution (b) is similar in position, colour and size to 100.0 mL with the mobile phase. Dilute 1.0 mL of this
theprincipal spot in the chromatogram obtained with solution to 10.0 mL with the mobile phase.
reference solution (a). Column:
D.lt gives reaction (a) of chlorides (2.3.1). - size: 1 = 0.15 m, 0 = 4 nun;
TESTS - stationary phase: O<tylsi/j'1 silica gelfor chromatography R
(5 urn).
Solution S
Dissolve 1.0 g in carbon dioxide-free waterR and dilute to Mobile phase Mix 0.656 g of sodium octanesulfonate R with
50 mL with the same solvent. 150 mL of acetonitrile R and dilute to 500 mL with
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2022 Alprostadil 1-113
phosphate buffer pH 2.8 prepared as follows: mix 1.78 g of r""y0H

phosphoric acidRand 15.6 g of sodium dihydrogen phosphate R
and dilute to 2000 mL with water R. ~CH,
Flow rate 1 mUmin.
B. 2-(prop-2-enyl)phenol,
Equilibration With the mobile phase for about 1 h.
Injection 20 1'1-.
and enanllomer
Run time Twice the retention time of alprenolol.
Retenlion lime Alprenolol = about 11 min;
4-isopropylphenol = about 18 min.
System suitability Reference solution (a): C. (2RS)-I-[ (I-methylethyl)amino1-3-[2-(prop-l-<:nyl)
- resolution: minimum 5 between the peaks due to alprenolol phenoXYlpropan-2-0~
and 4-isopropylphenol; if necessary) adjust the
concentration of sodium octanesulfcnate and/or
acetonitrile in the mobile phase (increase me
concentration of sodium octanesulfonate to increase the
retention time of alprenolol and increase the
concentration of acetonitrile to decrease the
retention times of both compounds). D. 1,1'-[( I-methylethyl)iminolbis[3-[2-(proI'"2-enyl)phenoXYl
Limits: propan-z-ol].
- unspecified impuruies: for each impurity) not more than _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE"
0.25 times the area of the principal peakin the
(0.10 per cent);
Alprostadil ***
chromatogram obtainedwith reference solution (b) *** ***
-
(0.4 per cent);
disregard limit: 0.1 times the area of the principal peak in
(ph. Bur. monogroph 1488) ***
(0.04 per cent).
vaaro.
Maximum 0.1 per cent, determined on 1.0 g. 354.5 745-65-3
ASSAY
Action and use
Dissolve 0.400 g in 25 mL of a mixrure of equal volumes of
Prostaglandin E, (pGE,).
anhydrous ethanol R and water R. Add 10 mL of 0.01 M
hydrochloric add. Carry out a potentiometric titration (2.2.20), PIlE" _
using 0.1 M sodium hydroxide. Read the volume added
between the 2 points of inflexion. DEFINITION
7- [( 1R,2R,3R)-3-Hydroxy-2-[(I B,3S)-3-hydroxyoct-I-enyt]-
I mL of 0.1 M sodium hydroxide is equivalent to 28.58 mg
5-oxocyclopentyllheptanoic acid.
of C I5H,.CINO,.
Content
STORAGE 95.0 per cent to 102.5 per cent (anhydrous substance).
CHARACTERS
IMPURITIES Appearance
Specified imputities C, D. White or slightly yellowish, crystalline powder.
Other detectable impurities (the following substances would, if Solublllty
present at a wjficientleveJ, be detected by one or other of the tests Practically insoluble in water, freely solublein ethanol
in the monograph. They are limited by thegeneral acceptance (96 per cent), soluble in acetone, slightly soluble in ethyl
criterion for other/unspecified impurities and/or try the general acetate.
monogroph Substances for pharmaceutical use (2034). It is
there/ore not neussary to idenuJy these impun'll"es for IDENTIFICATION
demonstration of compliance. See also 5.10. Control of impurities A. Specific optical rotation (2.2.7): -70 to -60 (anhydrous
in substanas for pharmaceutical use) A, B. substance).
Immediately before use, dissolve 50 mg in ethanol
H OH (96 per cenO R and dilute to 10.0 mL with the same solvent.
O~OH and enanOOmer
r""yv............... B. Infrared absorption spectrophotometry (2.2.24).
~CH2 Comparison alprostadil CRS.

C. Examine the chromatograms obtained in the assay.
A. (2RS)-3-[2-(proI'"2-enyl)phenoXYlpropan-I,2-diol, Results The principal peakin the chromatogram obtained
with the test solutionis similar in retention time and size to
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1-114 Alprostadil 2022
the principal peak in the chromatogram obtained with the System B

reference solution. Use me same conditions as for systemA with the following
TESTS mobile phase and elution programme:
- mobile phase A: dissolve 3.9 g of sodium dihydrogen
Related substances
phosphate R in water R and dilute to 1.0 L with the same
Liquid chromatography (2.2.29). Prepare the solutions protected
solvent; adjust to pH 2.5 with a 2.9 gIL solution of
from light.
phosphoric acidR (approximately 600 mL is required);
Test solution Dissolve 10.0 mg of the substance to be to 600 mL of the buffer solution add 400 mL of
examined in a mixture of equalvolumes of aceumitn"le Rl and acetonitrile R J;
water R and dilute to 10.0 mL with the same mixture of - mobile phase B: use mobile phase-B as described under
solvents. system A;
Reference solution (a) Dilute 100 ~L of the test solution to
20.0 mL with a mixture of equal volumes of acetonitrile Rl Thn' MobUe phase A Mobile phase B
and water R. (mIn) (per cent V/II) (per cent V/1?
Reference sdsuion (b) Dissolve 1.0 mg of dinoprouone 0·50 100 0
,mpuniy C CRS (a1prostadil impurity H) and 1.0 mg of the 50· 51 100 -i 0 0---> LOO
substance £0 be examined in a mixture of equal volumes of 51 - 61 0 100
ace«mitrile RJ and water R and dilute [Q 20.0 mL with the 61 - 62 0--->100 100 -i 0
same mixture of solvents. 62 - 72 '00 0
Reference solution (c) In oedec to prepare impurities A and B
in situ, dissolve 1 mg of the substance to be examined in Relativeretention With reference to alprostadil (retention
100 ul., of 1 M sodium hydroxide (the solution becomes time = about 7 min): impurity A = about 2.4;
brownish-red), wait for 3 min and add 100 ~L of a 112 gIL impurity B =about 2.6.
solution of p!wsphork acidR (yellowish-white opalescent System suilability:
solution); dilute to 5.0 mL with a mixture of equal volumes - resolution: minimum 1.5 between the peaksdue to
of acetonitrile RJ and water R. impurity A and impurity B in the chromatogram obtained
System A with reference solution (c).
Column: Carry out the test according to systemA and B.
- size: 1= 0.25 m, 0 = 4.0 mrn; Limits:
- stationary phase: bose-deactiuated octy/silyl silica gelfor - comaion factors: for the calculation of content) multiply
chromatography R (4 um) with a poce size of 6 nrn; the peak areas of the impurities listed in Table 1488.-1 by
- temperature: 35°C. the corresponding correction factor;
Mobile phase:
- mobile phase A: dissolve 3.9 g of sodium dihydrogen Table 1488.-1.
phosphate R in water R and dilute £0 1.0 L with the same hnpurlty Relative retention Relative retention Correction factor
solvent; adjust to pH 2.5 with a 2.9 gILsolution of (system A) (sY!ltem B)
phosphoric acidR (approximately 600 mL is required); impurity G 0.80 0.7
to 740 mL of the buffer solution add 260 mL of impurity F 0.88 0.8
aceumitrile RJj impurityD 0.90 1.0
- mobile phaseB: dissolve 3.9 g of sodium dihydrogen impurityH 0.96 0.7
phosphate R in water R and dilute £0 1.0 L with the same impurityB 1.10 0.7
solvent; adjust to pH 2.5 with a 2.9 gIL solution of impurity C 1.36 I.,
phosphoric acid R (approximately 600 mL is required); impurityK l.85 0.06
to 200 mL of the buffer solution add 800 mL of impurity A 2.32 0.7
acetonitrile Rl; impurity B 2.45 1.5
impurity.I 4.00 1.0
Thn' MobUe phase A Moblle phase B impurityJ 5.89 1.0
(mIn) (per cent VIV) (per cent VIV)
0-75 100 0 - impun"ty A: not more than 3 times the area of the
75 - 76 100 -i 0 o -i 100 principal peak in the chromatogram obtained with
76 - 86 0 100 reference solution (a) (1.5 per cent);
86·87 o -i 100 100 - i 0 - impurity B: not more than the area of the principal peak in
87 - 102 100 0 the chromatogram obtained with reference solution (a)
(0.5 per cent);
Flmo rare I mUmin. - any other impun'ty: not more than 1.8 times the area of the
Detection Spectrophotometer at 200 nm. principal peak in the chromatogram obtained with
reference solution (a) (0.9 per cent), and not more than 1
Injection 20~.
such peakhas an area greater than the area of the
Retention time AJprostadil = about 63 min. principal peak in the chromatogram obtained with
Systemsuitability: reference solution (a) (0.5 per cent). Evaluate impurities
- resolution: minimum 1.5 between the peaks due [0 appearing at relative retentions less than 1.2 by systemA
impurity Hand alprostadil in the chromatogram obtained and impurities appearing at relative retentions greater than
with reference solution (b). 1.2 by system B;
- LOud: not more than 3 times the area of the principal peak
(1.5 per cent);
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2022 Alprostadil 1-115

(0.05 per cent),
Wa'er (2.5.32)
Maximum 0.5 per cent) determined on 50 mg.
ASSAY E. 7-[(IR,2R,3S)-3-hydroxy-2-[(IE,3S)-3-hydroxyoct-l-
Liquid chromatography (2.2.29) as described in the test for enyl]-5-oxocyclopentyllheptanoic acid (11-
relatedsubstances) system A. .Prepare the solutions protected epiprostaglandin E I ) ,
from light.
examined in a mixture of equal volumes of acetonittiie RI and Co,H
water R and dilute to 25.0 mL with the same mixture of
solvents. Dilute 3.0 mL of the solution to 20.0 mL with a CH,
mixtureof equal volumes of acetonitrile RI and WQt.er R.
Reference solution Dissolve 5.0 mg of alpwstadil CRS in a
mixture of equal volumes of ace«m;tn"le Rl and water Rand F. 7-[(lS,2R,3R)-3-hydroxy-2-[(IE,3S)-3-hydroxyoct-l-
dilute to 25.0 mL with the same mixture of solvents. Dilute enyl]-5-oxocyclopentyl]hep,anoic acid
6.0 mL of the solution '0
20.0 mL with a mixture of (8-epiprostaglandin E I ) ,
equal volumes of acetonitrile Rl and water R.
Injeaion 20 pL. o
Calculate the percentage content of C2oH340S taking into
account the assigned content of alprostadi/ CRS.
CH,
STORAGE HO ".
At a temperature of 2 °C to 8 °C. H H OH
IMPURITIES G. (5Z)-7-[(IR,2R,3R)-3-hydroxy-2-[(IE,3S)-3-hydroxyOCl-
l-enyI1-5-oxocyclopentyl]hepr-5-enoic acid
(dinoprostone),
A. 7-[(1 R,2S)-2-[(IE.3S)-3-hydroxyoct-l-enyl]-5-
oxocydopent-3-enyl]heptanoic acid (prostaglandin AI),
o H. (5E)-7-[(IR,2R,3R)-3-hydroxy-2-[(IE,3S)-3-hydroxyoct-
CO,H l-enylj-5-oxocyclopentyl]hep'-5-enoic acid «5E)-
prostaglandin 1>,),
CH,
o o
H""'~O"""""""CH3
B. 7-[2-[(1E,3S) -3-hydroxyoc'-I-enyl]-s-oxocvclopenr-1-
CH,
enyljheptanoic acid (prostaglandin Bj),
HO'
H H OH
I. ethyl 7-[(IR,2R,3R)-3-hydroxy-2-[(IE,3S)-3-hydroxyoct-
l-enyl]-5-oxocyclopentyl]heptanoate (prostaglandin EI>
ethyl ester),
o
o o CH3
C.7-[(lR,2R,3R)-3-hydroxy-2-[(IE)-3-oxooct-l-enyl]-5- H"'~oAcH,
oxocyclopentyljheptanoic acid (Is-oxoprostaglendln E I ) ,
CH,
H H OH
J. I-methylethyl 7 -[(lR,2R.3R)-3-hydroxy-2-[(IE,3S)-3-
HO hydroxyoct-l-enyl]-5-oxocyclopentyl]heptanoate
(prostaglandin E I , isopropyl ester),
D.7-[(IR,2R,3R)-3-bydroxy-2-[(IE,3R)-3-hydroxyoct-l-
enyl]-5-oxocyclopentyllheplanoic acid (15-
epiprostaglandin E I ) ,
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1-116 Alteplase 2022
o If alteplase is stored in bulk form, stability (maintenance of
O-6~-O
P'I
potency) in the intended storage conditions must be
demonstrated.
The production, purification and product consistency are
"'" checked by a number of analytical methods described below,

carried out routinely as in-process controls.
K. triphenylphosphine oxide. Protein content
________ ~ PhE" The protein concentration of alteplase solutions is
determined by measuring the absorbance (2.2.25) of the
protein solution at 280 run and at 320 nm, using formulation
buffer as the compensation liquid. If dilution of alteplase
Alteplase for Injection samples is necessary, the samples are diluted in formulation
buffer. For the calculation of the alteplase concentration, the
(Ph. Eur. monograph 1170) absorbance value (A 280 - A J 20) is divided by the specific
absorption coefficient for alteplase of 1.9.
S~QVICRDEK TQHIYQQHQS WLRPVLRSNR .
VEYCWCNSGR Potency
.... ~HSVPVKS CSEPR9FNGG
.
TCQQALY~SD ,
FVCQCPEGFA
'
The potency of alteplase is determined in an in vitro clot-lysis
GKCCEIDTRA ~YEDQGrSY RGTWSTAESG AECTNWNSSA
assay as described under Assay. The specific activity of bulk
LAQl(PYSGRR PDAIRLGLGN HNY9RNPDRD ISf(~"'VFKA alteplase is approximately 580 000 IU per milligram of
alteplase. .
GK,{SSEF~ST PACSEGNSDC 'ifGNGSAYRG THSLTESGAS
N-terminal sequence
~pWNSMILI GKV,{T~QNP~QALGLGKHN Y9RNPDGDAK
N-tenninal sequencing is applied to determine the correct
PHCHVLKNRR LTHEYCDVPS CSTCGLRQYS QPQFR N-tenninal sequence and to determine semiquantitatively
IKGGL additional cleavage sites in the alteplase molecule, for
FADIASHPWQ AAIFAKHRRS PGERFLCGGI LISSCHILSA example at position AA 275-276 or at position AA 27-28.
I
AHCFQERFPP HHLTVILGRT YRv7PG~EEO J KFEv~KnvH The N-tenninal sequence must conform with the sequence of
human tissue plasminogen activator.
KEFDDDTYDN DIALLQLKSD SSRCAQESSV VRTVCLPPAD
Iso electric focusing
LQLPDHTECE LSGYGKHEAL SPFYSERLKE AHYRLYPSSR
The consistency in the microheterogeneity of glycosylation of
tTS~LLNRTJVTDNMLCA~D TRSGGPQANL
I
HDArQGDSGG
the alteplase molecule can be demonstrated by isoelectric
PLVCLNDGRH TLVGIISWGL GCGQI<DVPGV YTKVTNYLDH focusing (IEF). A complex banding pattern with 10 major
IRDNMRP and several minor bands in the pH range 6.5-8.5 is observed.
Denaturing conditions are applied to achieve a good
Action and use separation of differently charged variants of alteplase.
Tissue-type plasminogen activator; fibrinolytic. The broad charge distribution observed is due to a
population of molecules, which differ in the fine structure of
PhE" _
biantenary and triantenary complex-type carbohydrate
DEFINITION residues, with different degrees of substitution with sialic
Alteplase for injection is a sterile, freeze-dried preparation of acids. The banding pattern of alteplase test samples must be
alreplase, a tissue plasminogen activator produced by consistent with the pattern of alteplase reference standard.
recombinant DNA technology. It has a potency of not less Single-chain alteplase content
than 500 000 IU per milligram of protein. The alteplase produced by CHO (Chinese hamster ovary)
Tissue plasminogen activator binds to fibrin clots and cells in serum-free medium is predominantly single-chain
activates plasminogen, leading to the generation of plasmin alteplase. The single-chain fonn can be separated from the
and to the degradation of fibrin clots or blood coagulates. two-chain form by gel-permeation liquid chromatography
under reducing conditions as described under Single-chain
Alteplase consists of 527 amino acids with a calculated
content (see Tests). The single-chain alteplase content in
relative molecular mass of 59 050 without consideration of
bulk samples must he higher than 60 per cent,
the carbohydrate moieties attached at positions Asn 117,
Asn 184 and Asn 448. The total relative molecular mass is Tryptic-peptide mapping
approximately 65 000. Alteplase is cleaved by plasmin The primary structure of the alteplase molecule is verified by
between amino-acids 275 and 276 into a two-chain form (A tryptic-peptide mapping as described under Identification B.
chain and B chain) that are connected by a disulfide bridge The reduced and carboxymethylated molecule is cleaved by
between Cys 264 and Cys 395. The single-chain form and trypsin into about 50 peptides, which are separated by
the two-chain form show comparable fibrinolytic activity in reverse-phase liquid chromatography. A characteristic
vitro. chromatogram (fingerprint) is obtained. The identity of the
tryptic-peptide map of a given alteplase sample with the
PRODUCTION
profile of a well-characterised reference standard is an
Alteplase is produced by recombinant DNA synthesis in cell indirect confirmation of the amino-acid sequence, because
culture; the fermentation takes place in serum-free medium. even single amino-acid exchanges in individual peptides can
The purification process is designed to remove efficiently be detected by this sensitive technique. In addition, complex
potential impurities, such as antibiotics, DNA and protein peaks of the glycopeptides can be isolated from the tryptic-
contaminants derived both from the host cell and from the peptide map and separated in a second dimension, either by
production medium, and potential viral contaminants. reverse-phase liquid chromatography under modified
conditions or by capillary electrophoresis. By this two-
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2022 Alteplase 1-117
dimensional separation of glycopeptide variants) lot-to-lot and reference standard and using a relative molecular mass of
consistency of the microheterogeneity of glycosylation can be 180.2 for mannose and a relative molecular mass of 59 050
demonstrated. for the alteplase protein moiety. The neutral sugar content of
The tryptic-peptide map of alteplase samples must be the alteplase samples must be in the range of 70 to
consistent with the tryptic-peptide map of alteplase reference 130 per cent compared to alteplase reference standard, which
standard. contains about 12 moles of neutral sugar per mole of
alteplase.
Monomer content
The monomer content of alteplase is measured by gel- CHARACTERS
permeation liquid chromatography under non-reduced White or slightly yellow powder or solid friable mass.
conditions as described under Monomer content (see Tests). Reconstuute the preparation as stated on the label immediately
The monomer content of altepJase bulk samples must he before canying ou' the Identification, Tesu (except thosefor
higher than 95 per cent. solubility and water) and Assay.
Type VType II alteplase content IDENTIFICATION
CHO cells produce 2 glycosylation variants of alteplase. A. The assay serves aiso to identify the preparation.
Type I alteplase contains 1 pnlymannose-type gJycosylation at
B. Tryptic-peptide mapping. Examine by liquid
position Asn 117 and 2 complex-type glycosylation sites at
chromatography (2.2.29).
positions Aso 184 and Aso 448. Type Il alteplase is only
glycosylated at positions Aso 117 and Asn 448. Testsolution Dilute the preparation to be examined with
water R to obtain a solution containing about 1 mg of
The ratio of Type Iffype IT alteplase is constant in the range
alteplase per milliUtre. Dialyse about 2.5 mL of the solution
of 45 to 65 per cent of Type I and 35 to 55 per cent of
for at least 12 h into a solution containing 480 gIL of urea R,
Type II. The content of alteplase Type I and Type II can be
44 gIL of tris(hydroxymethyl)aminomethane R and 1.5 gIL of
determined by a densitometric scan of SDS-PAGE (sodium
sodium edeuue R and adjusted to pH 8.6, using a membrane
dodecyl sulfate polyacrylamide gel electrophoresis) gel.
with a cut-off point corresponding to a relative molecular
Plasmin-treated samples of alreplase, which are reduced and
mass of 10 000 for globular proteins. Measure the volume of
carboxymethylated before loading on the gel, are separated
the solution, transfer it to a clean test-tube and add per
into 3 bands: Type I alteplase A-chain (AA 1-275), Type II
millilitre 10 ~L nf a 156 gIL solutiou of dithiothreitol R. Allow
alteplase A-chain (AA 1-275) and alteplase B-chain
to stand for 4 h, cool in iced water and add per millilitre of
(AA 276-527). The ratio of Type Iffype II alteplase is
solution 25 ~L of a freshly prepared 190 gIL solution of
determined from a calibration curve, which is obtained by a
iodoacetic acidR. Allow {O stand in the dark for 30 min.
densitometric scan of defined mixtures of purified Type I
Add per millilitre 50 ~L of dithiothreitol solution to stop the
alteplase and Type II alteplase standards.
reaction. Dialyse for 24 h against an 8 gIL solution of
SDS-PAGE ammoniumhydrogen carbonate R. Add 1 part of trypsin for
SDS-PAGE (silver staining) is used to demonstrate purity of peptide mappingR to 100 parts of the protein and allow to
the alteplase bulk material and the integrity of the alteplase stand for 6 h ro 8 h. Repeat the addition of rrypsin and allow
molecule. For alteplase bulk samples, no additional protein to stand for a total of 24 h.
bands compared to reference standard or degradation
Reference solution Prepare as for the test solution using a
products must occur in SDS-PAGE gels at a loading amount
suitable reference standard instead of the preparation to be
of 2.5 J.lg alteplase protein per lane and a limit of detection of
examined.
5 ng per protein (BSA) band.
The chromatographic procedure may be carried out using:
Bacterial endotoxins (2.6.14) - a colwnn 0.1 m long and 4.6 mm in internal diameter
Less than I W per milligram of alteplase. packed with oc'adecylsilyl silica gd for chromatography R
Sialic acids (5 11m to 10 pm);
Proceed using a suitable validated method developed Mobile phaseA 8 gIL solution of sodium dihydrogen
according to general chapter 2.2.59. Glycan analysis of phosphate R, adjusted to pH 2.85 with phosphoric acidR,
glycoproteins. The sialic acids content for the test samples filtered and degassed;
must be in the range of 70 to 130 per cent compared to
Mobile phase B 75 per cent VIV solution of
alteplase reference standard, which contains about 3 moles of
acetonitrile R in mobile phase Aj
sialic acids per mole of alteplase.
- as detector a spectrophotometer set at 210 nm.
Neutral sugars Equilibrate the system with mobile phase A at a flow rate of
Dilute alteplase samples and the reference standard in the 1 mIJrnin. After injection of the solution, increase the
assay buller, containing 34.8 gIL of arginine R, 0.1 gIL of proportion of mobile phase B at a rate of 0.44 per cent per
polysorbate 80 R and adjusted to pH 7.4 with phosphoric minute until the ratio of mobile phase A to mobile phase B is
acid R, to a protein concentration of 50 J.lglmL. Prepare the 60:40, then increase the proportion of mobile phase B at a
following concentrations of mannose in the same assay buffer rate of 1.33 per cent per minute until the ratio of mobile
for a calibration curve: 20, 30, 40, 50 and 60 ~glmL. Pipette phase A to mobile phase B is 20:80 and then continue
2 mL of alteplase samples and reference standard, as well as elution with this mixture for a further 10 min. Record the
2 mL of each mannose concentration in duplicate in reagent chromatogram for the reference solution: the test is not valid
tubes. Add 50 ~L of phenolR, followed by 5 mL of sulfuric unless the resolution of peaks 6 (peptides 268-275) and 7
acid R, in each reagent tube. Incubate the mixture for 30 min (peptides 1-7) is at least l.5j WJrI and WIIZ are not more than
at room temperature..Measure the absorbance at 492 om for 0.4 min. Inject about 100 ~L of the test solution and record
each tube. Read the content of neutral sugars from the the chromatogram. Verify the identity of the peaks by
mannose calibration curve. The neutral sugar content is comparison with the chromatograms of the reference
expressed in moles of neutral sugar per mole of alreplase, solution. There should not be any additional significant peaks
taking into account the dilution factor for alteplase samples or shoulders, a significant peak or shoulder being defined as
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1-118 A1teplase 2022
100
80
::! \!!
60 \!!
..
~
n '$. e
~ ~
t:
-
40 N
.. e
..
e
~
\!!~
a
N
~
e
~ ..
20
o
1.1 1 lA. W. ft, V II.U cl J
20 40 60 80 100 120
Figure 1170.-1. - Chromatogram for tryptic-peptide mapping of altep/ase
one with an area response equal to or greater than 5 per cent - a column 0.6 m long and 7.5 mm in internal diameter
of peak 19 (peptides 278-296); no significant peak is missing. packed with silica-based, rigid, hydrophilic gel with
A type chromatogram for identification of the peaks cited is spherical particles 10 pm to 13 urn in diameter, suitable
shown in Figure 1170.-1. for size-exclusion chromatography;
- as mobile phase at a flow rate of 0.5 mUmin a solution
TESTS
containing 30 gIL of sodimn dihydrogen phosphore Rand
I gIL of sodium dodecyl sulfare R, adjusted to pH 6.8 with
The reconstituted preparation is clear (2.2.1) and not more
dilute sodium hydroxide solution R;
intensely coloured than reference solution Y, {2.2.2}
- as detector a spectrophotometer set at 214 nm.
Method II).
Inject about 50 ""Lof the test solution and record the
pH (2.2.3)
chromatogram. The chromatogram shows 2 major peaks
7.1 to 7.5.
corresponding to single-chain and two-cham alteplase.
Solubility Calculate the relative amount of single-chain alteplase from
Add the volume of the liquid stated on the label. the peak area values.
The preparation dissolves completely within 2 min at 20°C The test is not valid unless: the number of theoretical plates
to 25 "C, calculated on the basis of the single-chain alteplase peak is at
Protein content least 1000. The content of single-chain alteplase is not less
Prepare a solution of the substance to be examined with an than 60 per cent of the total amount of alteplase-related
accurately known concentration of about 1 gIL. Using a substances found.
34.8 gIL solution of arginine R adjusted to pH 7.3 with Monomer content
phosphoric add R, dilute an accurately measured volume of Examine by liquid chsomatography (2.2.29).
the solution of the substance to be examined so that the
absorbance measured at the maximum at about 280 om is
Test solution Reconstitute the preparation to be examined to
obtain a solution containing about 1 mg per millilirre,
0.5 to 1.0 (test solution). Measure the absorbance (2.2.25) of
the solution at the maximum at about 280 nm and at The chromatographic procedure may be carried out using:
320 run using the arginine solution as the compensation - a column 0.6 m long and 7.5 mm in internal diameter
liquid. Calculate the protein content in the portion of packed with silica-based rigid, hydrophilic gel with
alteplase taken from the following expression: spherical particles 10 ""01 to 13 JIm in diameter) suitable
for size-exclusion chromatography;
V(A 2so - An.) - as mobile phase at a flow rate of 0.5 mUmin a solution
1.9 containing 30 gIL of sodium dihydrogen phosphore R and
I gIL of sodium dodecyl sulfate R, adjusted to pH 6.8 with
in which V is the volume of the test solution, Azgo is the dilute sodium hydroxide solution R;
absorbance at the maximum at about 280 run and A 320 is the - as detector a spectrophotometer set at 214 nm.
absorbance at 320 run. Inject the test solution and record the chromatogram.
Single-chain content The test is not valid unless the number of theoretical plates
Examine by liquid chsomatography (2.2.29). calculated for the alreplase monomer peak is at least 1000.
Test solution Dissolve the preparation to be examined in Measure the response for all peaks) i.e, peaks corresponding
water R to obtain a solution containing about 1 mg of to alteplase species of different molecular masses. Calculate
alteplase per millilitre. Place about J mL of the solution in a the relative content of monomer from the area values of these
tube, add 3 mL of a 3 gIL solution of dithiothreiU!1 R in the peaks. The monomer content for alteplase must be at least
mobile phase, place a cap on the tube and heat at about 95 per cent.
80°C for 3 min to 5 min. Water (2.5.12)
The chromatographic procedure may he carried out using: Not more than 4.0 per cent, determined by the semi-micro
determination of water.
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2022 Altizide 1-119
Bacterial endotoxins (2.6.14) [(IoglO') - oj

Less than I IU per milligram of protein. b
Sterility (2.6.1) Calculate the aIteplase activity in International Units per
It complies with the test for sterility. millilitre from the following expression:
ASSAY DxUA
The potency of alteplase is determined by comparing its
abilityto activate plasminogen to form plasmin with the same in which D is the dilution factor for the test solution.
capacityof a reference preparation calibrated in International Calculate the specific activity in the portionof the substance
Units. The formation of plasmin is measured by the to be examined from the following expression:
determination of the lysis time of a fibrin clot in given
conditions. UA
The International Unit is the activity of a stated quantity of P
the International Standard of alteplase. The equivalence in in which P is the concentration of proteinobtained in the lest
International Units of the International Standard is stated by for proteincontent.
the World Health Organization. The estimated potency is not less than 90 per cent and not
Solvent buffer A solution containing 1.38 gIL of sodium more than 110 per cent of the stated potency.
dihydrog,n phosphate monohydrate R, 7.10 gIL of anhydrous
disodium hydrogen phosphat< R, 0.20 gIL of sodium azide Rand STORAGE
0.10 gIL of polysorbat< 80 R. Store in a colourless, glass container, under vacuum or under
an inerr gas, protected from light, at a temperature of 2 °C to
Human thrombin solution A solutionof human thrombin R
30°C.
containing 33 IU/mL in solventbuffer.
Human fibrinogen solution A 2 gILsolution ofjibn·nogen R in LABELLING
solvent buffer. Thelabelsuues:
- the number of International Units per container;
Human plasminogen sdution A 1 gIL solution of human
- the amount of protein per container;
plasminogen R in solvent buffer.
- the name and volume of the liquid to be used for
Test solutions Using a solution of the substance (Q be reconstitution.
examined containing I gI4 prepare serial dilutions using _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
solvent buffer, for example 1:5000, 1:10 000, 1:20000.
Reference solutions Using a solution of a suitable reference
standard havingan accurately known concentration of about
I gIL (580 000 IU of alteplase per millilitre), prepare 5 serial
dilutions using water R to obtain reference solutionshaving Altizide
known concentrations in the range 9.0 IU/mL to 145 IU/mL.
To each of a set of labelled glass test-tubes, add 0.5 mL of
human thrombin solution. Allocateeach test and reference
solution to a separate tube and add to each tube 0.5 mL of
and enanliomer
the solution allocated [0 it. To each of a second set of
labelled glass tubes, add 20 ~L of human plasminogen
solution, and I mL of human fibrinogen solution, mix and
store on ice. Beginning with the reference/thrombin mixture 383.9 5588-16-9
containing the lowest nwnber of International Units per
millilitre, record the time and separately add 200 ~L of each Action and use
of the thrombin mixtures to the test tubes containing the Thiazide diuretic.
plasminogen-fibrinogen mixture. Using a vortex mixer,
PhE" _
intermittently mix the contents of each tube for a total of
15 s and carefully place in a rack in a circulating water-bath DEFINmON
at 37 °C. A visibly turbid clot forms within 30 s and bubbles (3RS)-6-Chloro-3-[(prop-2-enylsulfanyl)methyl]-3,4-dihydro-
subsequently fonn within the clot. Record the clot-lysis time 2H-l)2,4-benzothiadiazine-7-sulfonamide I,I-dioxide.
as the time between the first addition of alteplase solution
and the moment when the last bubblerises to the surface. Content
Using a least-squares fit, determine the equation of the line 97.5 per cent to 102.0 per cent (anhydrous substance).
using the logarithms of the concentrations of the reference CHARACTERS
preparation in International Units per millilitre versus the Appearance
logarithms of the values of theirclot-lysis times in seconds, White or almost white powder.
according to the following equation:
Solubility
loglO' ~ a + b(logIOU,) Practically insoluble in water, soluble In-methanol, practically
insoluble in methylene chloride.
in which r is the clot-lysis time, Us the activity in It shows polymorphism (5.9).
International Units per rmllilitre of the reference preparation,
IDENTIFICATION
b is the slope and a they-intercept of the line. The test is not
valid unless the correlation coefficient is -0.9900 to -1.0000.
From the line equation and the clot-lysis time for the [est Comparison altizide CRS.
solution) calculate the logaritlun of the activity VA from the If the spectra obtainedshow differences, dissolve 50 mg of
following equation: the substance to be examined and 50 mg of the reference
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1-120 Alum 2022
substance separately in 2 mL of acetone R and evaporate the System sur'labi/ity Reference solution (c):
solvent. Precipitate by adding 1 mL of methylene chloride R. - resolution: minimum 1.0 between the peaks due to altizide
Evaporate to dryness and record Dew spectra using the and furosemide.
residues. Limie:
TESTS - impurity A: not more than 3 times the area of the
Impurity B principal peak in the chromatogram obtained with
Thin-layer chromatography (2.2.27). reference solution (a) (0.3 per cent);
Test solution Dissolve 0.200 g of the substance to be - unspedfied impurities: for each impurity, not more than the
examined in acetene R and dilute to 2.0 mL with the same area of the principal peak in the chromatogram obtained
solvent. with reference solution (a) (0.10 per cent);
Reference solution (a) Dissolve 10.0 mg of altizide
impurity B CRS in acerone R and dilute to 25.0 mL with the
(0.5 per cent);
same solvent.
Reference solution (b) To 1.0 mL of reference solution (a) the chromatogram obtained with reference solution (a)
add 1.0 mL of the test solution. (0.05 per cent).
Reference solution (c) Dilute 5.0 mL of reference solution (a)
to 10.0 mL with acerone R. Water (2.5.32)
Maximum 0.5 per cent, determined on 50.0 mg.
Plate TLC silica gel F", plate R.
Mobile phase acerone R, methylene chloride R (25:75 VII'). Sulfated ash (2.4. J 4)
Application 10 ilL of the test solution and reference
solutions (b) and (c). ASSAY
Deodopmem Over 2/3 of the plate. Liquid chromatography (2.2.29) as described in the test for
Drying In air. related substances, with the following modifications.
Detection Spray with a mixture of equal volumes of a 10 gIL Test solution Dissolve 25.0 mg of the substance to be
solution of potassium pennanganate R and a 50 gIL solution of examined in 2 rnL of acetonitrile R and dilute to 25.0 mL
sodium carbonate R, prepared immediately beforeuse. Allow with the mobile phase.
to stand for 30 min and examine in daylight. Reference solntien Dissolve 25.0 mg of altizide CRS in 2 mL
System suitability Reference solution (b): of acetonlink R and dilute to 25.0 mL with the mobile phase.
- the chromatogram shows 2 clearly separated spots. Calculate the percentage content of CIIHI4ClN30ot,S3 from
Limit Any spot due to impurity B is not more intense than the declared content of altizide CRS.
the principal spot in the chromatogram obtained with
IMPURITIES
reference solution (c) (0.2 per cent).
Specified imp""'tres A, B.
Related substances
Liquid chromatography (2.2.29). Prepare 'he solutions
immediately before use, except riference solution (b).
Test solution Dissolve 50 mg of the substance to he
examined in 5 mL of acetonitrile R and dilute to 25 mL with
the mobile phase.
Reference solUMn (a) Dilute 1.0 mL of the test solution to A. 4-amino-6-chlorobenzene-l,3-disulfonamide,
Reference soluuon (b) In order to produce impurity A in situ,
dissolve 50 mg of the substance to be examined in 5 mL of
aceronitrile R and dilute to 25 mL with water R. Allow to B. 3-[(2,2-diroethoxyethyl)sulfanyl]prop-I-ene.
stand for 30 min. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'I>E"
Reference solution (c) Dissolve 4 mg ofjurosemide CRS in

2 mL of acetonitrile R, add 2 mL of the test solution and
dilute to 100 mL with the mobile phase.
Column: Alum
- size: I =0.15 m, 0 = 3.9 mm;
- stationary phase: end-capped oetadeeylsilyl silica gelfor Potash Alum
chromatography with embedded poIor groups R (5 pm); Aluminium Potassium Sulphate
- temperature: 30°C. Aluminium Potassium Sulfate
Mobile phase acetonitrile R, water for chromatography R (Ph. Bur. monograph 0006)
previously adjusted to pH 2.0 with perchlone acidR
AlK(SO.l,,12H,O 474.4 7784-24-9
(25:75 VII').
Flow rate 0.7 mUmin. Action and use
Detection Spectrophotometer at 270 om. Astringent.
Injection 5 ~L. I'I>Ew _
Run time Twice the retention time of altizide.
DEFINITION
Rdative retention With reference to altizide (retention
Content
time = about 25 min): iropurity A = about 0.15;
99.0 per cent to 100.5 per cent of AlK(SO.l,,12H,O.
furosemide = about 1.05.
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2022 Aluminium Glycinate 1-121
CHARACTERS IDENTIFICATION
Appearance A. Dilute 0.1 mL of solution S2 (see Tests) to 2 mL with
Granular powder or colourless, transparent, crystalline water R. The solution gives reaction (a) of chlorides (2.3.J).
masses. B. Dilute 0.3 mL of solution 52 to 2 mL with water R.
Solubility The solution gives the reaction of aluminium (2.3.1).
Freely soluble in water, very soluble in boiling water, soluble TESTS
in glycerol, practically insoluble in ethanol (96 per cent). Solution SI
IDENIlFICATION Dissolve 10.0 g in distilled waterR and dilute to 100 mL with
A. Solution S (see Tests) gives the reactions of sulfates the same solvent.
(2.3.1). Solution S2
B. Solution S gives the reaction of aluminium (2.3.1). Dilute 50 mL of solution 81 to 100 mL with water R.
C. Shake 10 mL of solution S with 0.5 g of sodium hydrogen Appearance of solution
carbonau R and filter. The filtrate gives reaction (a) of Solution 82 is clear (2.2. J) and not more intenselycoloured
potassium (2.3.1). than reference solution B7 (2.2.2) Method II).
TESTS Sulfates (2.4.13)
Solution S Maximum 100 ppm, determined on solution 51.
Dissolve 2.5 g in water R and dilute to 50 mL with the same Iron (2.4.9)
solvent. Maximum 10 ppm, determined on solution 51.
Appearance of solution Alkali and alkaline-earth metals
Solution S is clear (2.2.1) and colourless (2.2.2, Method11). Maximum 0.5 per cent.
pH (2.2.3) To 20 mL of solution S2 add 100 mL of water R and heat to
3.0 to 3.5. boiling. To the hot solution add 0.2 mL of methylred
Dissolve 1.0 g in carbon dioxide-free water R and dilute to solution R. Add dilute ammonia R1 until the colour of the
10 mL with the same solvent. indicator changes to yellow and dilute to 150 mL with
Ammonlwn (2.4.1) water R. Heat to boiling and filter. Evaporate 75 mL of the
Maximum 0.2 per cent. filtrate to dryness on a water-bath and ignite to constant
To I mL of solution S add 4 mL of water R. Dilute 0.5 mL mass. The residue weighs a maximwn of 2.5 mg.
of this solution to 14 mL with water R. Water (2.5.12)
Iron (2.4.9) 42.0 per cent to 48.0 per cent, determined on 50.0 mg.
Maximum 100 ppm. ASSAY
Dilute 2 mL of solution S to 10 mL with water R. Use in this Dissolve 0.500 g in 25.0 mL of water R. Carry out the
test 0.3 mL of rhioglycaYic acidR. complexometric titration of aluminium (2.5.11). Titrate with
0.1 M zinc sulfate until the colour of the indicator changes
ASSAY
from greyish-green to pink. Cany out a blanktitration.
Dissolve 0.900 g in 20 mL of water R and carry out the
complexometric titration of aluminium (2.5.11). I mL of 0.1 M sodium <delate is equivalent to 24.14 mg
of AlCI,,6H 2 0 .
I mL of 0.1 M sodium <delate is equivalent ro 47.44 mg
of A1K(SO.h,12H2 0 . STORAGE
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" In an airtight container.
__________ ~ PhE"
Aluminium Chloride Hexahydrate

Aluminium Glycinate
(ph. Bur. managraph 0971)
AlCI,,6H 20 241.4 7784-13-6 NH2
£
\ /OH
AI .xH20
Action and use
/ 'OH
Astringent. o 0
Preparation
Aluminium Chloride Solution
G..f4AINO.,xH20 135.1 41354-48-7
PhE" _
Action and use
DEFINITION Antacid.
Content
95.0 per cent to 101.0 per cent. DEFINITION
CHARACTERS Aluminium Glycinate is a basic aluminium monoglycinate,
Appearance partly hydrated. It contains not less than 34.5% and not
White or slightly yellow, crystalline powder or colourless more than 38.5% of Ah03 and not less than 9.9% and not
crystals, deliquescent. more than 10.8% of N, both calculated with reference to the
dried substance.
Solubility
Very soluble in water, freely soluble in ethanol (96 per cent), CHARACTERISTICS
soluble in glycerol. A white or almost white powder.
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1-122 Aluminium Hydroxide 2022
Practically insoluble in water and in organic solvents.

It dissolves in dilutemineral acids and in aqueous solutions
Hydrated Aluminium Hydroxide for
of the alkali hydroxides. Adsorption
IDENTIFICATION (Ph. Eur. monograph 1664)
A. Add 0.1 g to 10 rnL of a solution prepared by dissolving
[AlO(OHl),xH,O
0.84 g of ciuic arid in 8 rnL of 1M sodium hydroxide and ""or ~
diluting to 20 rnL with water. Add 0.5 mL of a 0.1 % wlv

solutionof ninhydn"n in methanol and warm. A purple colour DEFINITION
is produced. Content
B. Suspend I g in 25 mL of O.5M hydrochlori< aeid and heat 90.0 per cent to 110.0 per cent of the content of aluminiwn
gentlyuntil a clearsolution is produced. Reserve half of the stated on the label.
solution. To 2 mL of the solution add 0.15 rnL of liquefied NOTE: shake the gelvigorously for at least 30 s immediately
phenol, shake and add carefully without shaking 5 mL of before examining.
dilute sodium hYfJO'hlon"te solution. A blue colouris produced. CHARACTERS
C. The solution reserved in test B yields the reaction Appearance
characteristic of aluminium salts, Appendix VI. White or almost white, translucent, viscous, colloidal gel.
TESTS A supernatant may be formed upon standing.
Acidity or alkalinity Solubility
pH of a suspension of I g in 25 rnL of carbon dioxide-free A clearor almostclearsolutionis obtained with alkali
water, 6.5 to 7.5, Appendix V L. hydroxide solutions and mineral acids.
Neutrallsing capacity IDENTIFICATION
Shake 0.2 g vigorously with 25 rnL of O.IM hydrochlori< acid Solution S (see Tests) givesthe reaction of aluminium.
for 5 minutes and allow to stand for 5 minutes. The pH of To 10 rnL of solution S add about 0.5 mL of dilute
the mixture is greater than 3.0, Appendix V L hydrochlori< aeid R and about 0.5 rnL of thioaatamide
Arsenic reagent R. No precipitate is formed. Add dropwise 5 mL of
Dissolve 2.0 g in 18 mL of brominated hydrochlori< arid and dilute sodium hydroxide solution R. Allow to standfor 1 h.
32 mL of water. 25 mL of the resulting solutioncomplies A gelatinous white precipitate is formed whichdissolves upon
with the limit testfor arsenic, Appendix vn (I ppm). addition of 5 mL of daute sodium hydroxide solution R.
Mercuric salts Gtadually add 5 rnL of ammonium chloride solution R and
Dissolve 2.0 g in 10 mL of 1M sulfuric add, transfer to a allow to standfor 30 min. The gelatinous whiteprecipitate is
separating funnel with the aid of water, dilute 10 about re-formed,
50 rnL with water and add 50 mL of 0.5M ndfune acid. TESTS
Add 100 rnL of water, 2 g of hydroxylamine hydrochloride, Solution S
I rnL of 0.05M disodium edetate and I rnL of glacial acetic Add I g to 4 rnL of hydrochloric aeid R. Heat at 60 "C for
acid. Add 5 mL of chlorofonn, shake, allow to separate and 1 h, cool, dilute to 50 mL with distilled water R and filter if
discard the chlorofonn layer. Titrate the aqueous layer with a necessary.
solution of dilhizone in chlorofonn containing 8 JIg per mL
pH (1.1.3)
until the chloroform layer remains green. After each addition,
5.5 to 8.5.
shake vigorously, allow the layers to separate and discard the
chloroform layer. Repeat the operation using a solution Adsorption power
prepared by diluting I rnL of mercury standard solution Dilute the substance to be examined with distilled water R to
(5 ppm Bg) to 100 rnL with 0.5M sulfuric aeid and beginning obtain an aluminium concentration of 5 mglmL. Prepare
at the words CAdd 100 mL of water. . . '. The volwne of the bovine albumin R solutions with the following concentrations
dithizone solution required by the substance being examined of bovine albumin: 0.5 mglmL, I mglmL, 2 mglmL,
does not exceed thatrequired by the mercury standard 3 mglmL, 5 mglrnL and 10 mglrnL. If necessary, adjust the
solution. gel and the bovine albumin R solutions to pH 6.0 with dilute
hydrochloric aeid R or dilute sodium hydroxide solution R.
Chloride
For adsorption; mix 1 partof the diluted gel with 4 parts of
Dissolve 1.0 g in 10 mL of 2M ninic acid and dilute to
each of the solutions of bovine albumin R and allow to stand
100 mL with water. 15 mL of the resulting solution complies
at room temperature for 1 h. During this time shake the
with the limit testfor chlorides, Appendix vn (330 ppm).
mixture vigorously at least 5 times. Centrifuge or filter
Loss on drying through a non-protein-retaining filter. Immediately determine
When dried to constant weight at 130°, loses not more than the protein Content (1.5.33, Method 1) of either the
12.0% of its weight. Use I g. supernatant or the filtrate.
ASSAY It complies with the test if no bovine albwnin is detectable in
ForA1,03 the supernatant or filtrate of the 2 mglmL bovine albumin R
Dissolve 0.25 g in a mixture of 3 rnL of 1Mhydrochlori< acid solution (maximum level of adsorption) and in the
and 50 rnL of water, add 50 rnL of 0.05M disodium edetate VS supernatant or filtrate of bovine albumin R solutions of lower
and neutralise with / M sodium hydroxide using methyl red concentrations. Solutions containing 3 mglmL, 5 rng/mf.and
solution as indicator. Heat the solution to boiling, allow to 10 mglmL bovine albumin R may show bovine albumin in the
stand for 10 minutes on a water bath, cool rapidly, add supernatant or filtrate, proportional to the amount of bovine
about 50 mg of xylenol orange triturate and 5 g of hexamine albumin in the solutions.
and titrate the excessof disodium edetate with O.05M lead Sedimentation
nitrate VS until the solution becomes red. EachmL of O.05M If necessary, adjust the substance to be examined to pH 6.0
disodium edetate VS is equivalent to 2.549 mg of A120J • using dilute hydrochlori< acid R or dilute sodium hydroxide
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2022 Aluminium Hydroxide 1-123
solution R. Dilute with distilled waterR to obtain an

aluminium concentration of approximately 5 mglmL. If the
Dried Aluminium Hydroxide
aluminium content of the substance to be examined is lower (Hydrated Aluminium Oxide.. Ph. Bur. monograph
than 5 mglmL, adjust to pH 6.0 and dilute with a 9 gIL 0311)
solution of sodium chloride R to obtain an aluminium
concentration of about 1 mglmL. After shaking for at least Action and use
30 s, place 25 mL of the preparation in a 25 mL graduated Antacid.
cylinder and allow to stand for 24 h. Preparations
It complies with the test if the volume of the clear Aluminium Hydroxide Chewable Tablets
supernatant is less than 5 mL for the gel with an aluminium Aluminium Hydroxide Oral Suspension
content of about 5 mglmL. Compound Magnesium Trisilicate Chewable Tablets
It complies with the test if the volume of the clear Co-magaldrox Oral Suspension
supernatant is less than 20 mL for the gel with an aluminium Co-magaldrox Tablets
content of about 1 mglmL.
1'1>£" _
Chlorides (2.4.4)
Maximum 0.33 per cent. DEFINITION
Dissolve 0.5 g in 10 mL of dilute nitric add R and dilute to Content
500 mL with water R. 47.0 pet cent to 60.0 per cent of Al 20, (M, 102.0).
Nitrates CHARACTERS
Place 5 g in a test-tube immersed in ice-water, add 0.4 mL White or almost white, amorphous powder.
of a 100 gIL solution of potassium chloride R, 0.1 mL of Solubility
diphenylamine solution Rand, dropwise with shaking, 5 mL of PracticaUy insoluble in water. It dissolves in dilute mineral
sulfuric acidR. Transfer the tube to a water-bath at 50 'C. acids and in solutions of alkali hydroxides.
After 15 min, any blue colour in the solution is not more IDENTIFICATION
intense than that in a standard. prepared at me same time Solution S (see Tests) gives the reaction of aluminium
and in the same manner using 5 mL of nitrate standard (2.3.1).
solUlion (100 ppm NO,) R.
TESTS
Sulfates (2.4.13)
Soludon S
Maximum 0.5 per cent. Dissolve 2.5 g in 15 mL of hydrochloric acid R, heating on a
Dilute 2 mL of solution S to 20 mL with waterR. water-bath. Dilute to 100 mL with distilled waterR.
Ammonium (2.4.1, Method H) Appearance of solution
Maximum 50 ppm, determined on 1.0 g. Solution S is not more opalescent than reference
Prepare the standard using 0.5 mL of ammonium standard suspension II (2.2.1) and not more intensely coloured than
solution (100 ppm NH.,) R. reference solution GY. (2.2.2, Method 11).
Arsenic (2.4.2, Method A) Alkaline impurities
Maximum 1 ppm, determined on 1 g. Shake 1.0 g with 20 mL of carbon dioxide-free waterR for
Iron (2.4.9) I min and filter. To 10 mL of the filtrate add 0.1 mL of
Maximum 15 ppm, determined on 0.67 g. phenolphthalein solution R. Any pink colour disappears on the
addition of 0.3 mL of 0.1 M hydrochlori< acid.
Bacterial endotoxins (2.6.14)
Less man 5 IV of endotoxin per milligram of aluminium, if Neutrallsing capacity
intended for use in the manufacture of an adsorbed product Carry out the rest at 37°C Disperse 0.5 gin 100 mL of
without a further appropriate procedure for the removal of water R, heat, add 100.0 mL of 0.1 M hydrochloric acid,
bacterial endotoxins. previously heated, and stir continuously; the pH (2.2.3) of
the solution after 10 min, 15 min and 20 min is not less than
ASSAY 1.8.. 2.3 and 3.0 respectively and is at no time greater than
Dissolve 2.50 g in 10 mL of hydrochloric acid R, heating for 4.5. Add 10.0 mL of 0.5 M hydrochlori< acid, previously
30 min at 100°C on a water-bath. Cool and dilute to 20 ml, heated, stir continuously for 1 h and titrate with O.11W
with water R. To 10 mL of the solution, add concentrated '0
sodium hydroxide pH 3.5; no' mOle than 35.0 mL of 0.1 M
ammonia R until a precipitate is obtained. Add the smallest sodium hydroxide is required.
quantity of hydrochlO1U acid R needed to dissolve the Chlorides (2.4.4)
precipitate and dilute to 20 mL with waterR. Carry out the
Maximum 1 per cent.
complexometric titration of aluminium (2.5.11). Carry out a
blank titration. Dissolve 0.1 g with heating in 10 mL of dilute nitric acidR
and dilute to 100 mL with water R. Dilute 5 mL of the
STORAGE solution to 15 mL with waterR.
At a temperature not exceeding 30°C. Do not allow to Sulfates (2.4.13)
freeze. If the substance is sterile, store in a sterile, airtight,
Maximum 1 per cent.
tamper-evident container.
Dilute 4 mL of solution S to 100 mL with distiUed waterR.
LABELLING
Arsenic (2.4.2, MethodA)
The label states the declared content of aluminium.
Maximum 4 ppm, determined on 10 mL of solution S.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'1>£"
TAMC: acceptance criterion 10' CFUlg (2.6.12).
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1-124 Aluminium Magnesium Silicate 2022
TYMC: acceptance criterion 102 CFUlg (2.6.12). CHARACTERS

Absence of bile-tolerant gram-negative bacteria (2.6.13). Appearance
Absence of Escherichia coli (2.6. H). Almost white, coarse powder, granules or flakes (types lA, IC
and IIA); almost white, fine powder (type IB).
ASSAY
Dissolve 0.800 g in 10 mL of hydrochloric acid R1, heating on Solubility
a water-bath. Cool and dilute to 50.0 mL with water R. Practically insoluble in water and in organic solvents.
To 10.0 mL of the solution add dilute ammonia Rl until a It swells in water to produce a colloidal dispersion.
precipitate begins to appear. Add the smaUest quantity of IDENTIFICATION
dilute hydrochlom acid R needed to dissolve the precipitate Carry out either tests A, B, C, F or tests D, E.
and dilute to 20 mL with water R. Carry out the
A. In a platinum crucible mix 1.0 g with 5.0 g of anhydrous
complexometric titration of aluminium (2.5.11).
lithium metaborate R. Heat slowly at first and ignite at
I mL of 0.1 M sodium edetate is equivalentto 5.098 mg 1000-1200"C for 15 min. Allow to cool and crush the
of AJ,O,. residue. 0.25 g of the residue gives the reaction of silicates
STORAGE (2.3.1).
In an airtight container, at a temperature not B. Dissolve 1.0 g of the residue obtained in identification
exceeding 30°C. test A in a mixture of 5 mL of dilute hydrochloric acidR and
FUNCTIONALITY-RELATED CHARACTERISTICS 10 mL of water R. Filter to obtain a clear solution and add
This section provides in/o1711ation on characteristics that an! ammonium chloride buffer solution pH 10.0 R. A white>
recognised as being relevant control parameters for oneor more gelatinous precipitate is formed. Centrifuge and keep the
funaions of the substance when usedas an excipient (see chapter supernatant for identification test C. Dissolve the precipitate
5.15). Some of the characteristics described in the Functionality- in dilute hydrochloric acidR. Add dropwise dilute sodium
related chamaenstia section may also bepresent in themandah»y hydroxide solUlion R. A white gelatinous precipitate is formed.
part of the monograph since they also represent mandatory quality Filter and add a few drops of phenolphlila1ein ,oI",umR to the
crueria. In such cases} a cross-reference to the tests descn·bed in the residue. The residue turns pink. Wash the residue with
mandatory part is included in the Functionality-related water R until the pink colour is completely discharged and
characteristics seeMn. Control of the characteristics can contribute the residue remains white upon addition of a drop of
to the quality of a medicinal produa by improving tire consistency phenolphthalein solution R. Sprinkle a few crystals of sodium
of the manuja£tun·ng process and theper/onnanu of the medicinal fluoride R on the residue. The residue, in contact with the
product during use. Where amtrol methods are cited, they are crystals, turns pink again in a short time.
reaJgnised as being suitable for'the purpose, but other methods can C. To 2 mL of the supernatant obtained aftercentrifugation
also be used. Wherever results for a particular charaaerudc are in identification test B, add 1 mL of dilute ammonia Rl and
reponed, the amtrolmethod must be indicated. 1 mL of ammonium chloride so/mien R. Upon the addition of
Thefollowing characteristics m'lY be relevant for hydrated dilute ammonia R1 a white precipitate may form, which
aluminium oxide usedas adsorbent. dissolves afteraddition of the ammonium chloride solution R.
Add I mL of disadium hydrogen pho,phare ,oIution R. A white
Particle-size distribution (2.9.31) precipitate is formed.
Specific surface area (2.9.26) D. X-ray diffraction (2.9.33), oriented sample.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Add 2 g in small portions to 100 mL of water R, with
vigorous shaking. Allow to stand for at least 12 h to ensure
complete hydration. Place 2 mL of the resulting mixture on a
Aluminium Magnesium Silicate suitable glass slide and allow to dry in air at room
temperature to produce an oriented film. Place the slide in a
(ph. Bur. monograph 1388) vacuum desiccatorover ethylene glycol R. Evacuate the
Action and use desiccator and close the stopcock so that the ethylene glycol
Excipient. saturates the chamber. Allow to stand for at least 12 h.
Record the X-ray diffraction pattern and calculate the
PIlE" ~ d values: the largest peak corresponds to a d value between
1.5 nm and 1.72 nm,
DEFINITION
Mixtureof colloidal-sizeparticles of montmorillonite and E. X-ray diffraction (2.9.33), random sample.
saponite, practically free from grit and non-swellable ore. Prepare a random powdersample, record the X-ray
The requirements for viscosity and ratio of aluminium diffraction pattern and determine the d values in the region
content to magnesium content differ for the several types of between 0.148 nm and 0.154 nm. Peaks are found between
aluminium magnesium silicate, as shown in the table below. 0.1492 nm and 0.1504 nm and between 0.1510 nm and
0.1540 nm.
F. It complies with the limits of the assay.
Viscosity (mPa-s) AI content J Mg content
Typo TESTS
min. max. min. max.
pH (2.2.3)
IA 225 600 0.5 1.2 9.0 to 10.0.
IB 150 450 0.5 1.2
Disperse 5.0 g in 100 mL of carbon dioxide-free water R.
Ie
Viscosity (2.2.1 U)
soo 2200 0.5 1.2
Weigh a quantity of the substance to be examined equivalent
l1A 100 300 1.4 2.S to 25.0 g of the dried substance and immediately transfer to
a suitable 1 L blender jarcontaining a quantity of waterR, at
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2022 Aluminium Magnesium Silicate 1-125
25 ± 2°C, that is sufficient to produce a mixture weighing Lead

500 g. Blend for exactly 3 min, at 14 000-15 000 r/min. Maximum 15 ppm.
The heat generated during blending causes the temperature Atomic absorption spectrometry (2.2.23, Me/lwd 1).
to rise to above 30°C. Transfer the contents of the blender
Test solution Transfer 10.0 g to a 250 mL beaker containing
to a 600 mL beaker and allow to stand for 5 min. 100 mL of dilute hydrochloric acidR. Mix, cover with a watch
The sample temperature should be 33 ± 3°C. glass and boil for 15 min. Allow to cool to room temperature
Using a suitable rotating viscometer equipped with a spindle and allow the insoluble matter to settle. Decant the
as specified below, operate the viscometer at 60 r/min for supernatant through a rapid-flow filter paper into a 400 mL
exactly 6 min and record the scale reading. beaker. To the insoluble matter in the 250 mL beaker add
For type IA, use a spindle with a cylinder 1.87 em in 25 mL of hot water R. Stir, allow the insoluble matter to
diameter and 0.69 em high attached to a shaft 0.32 em in settle and decant the supernatant through the filter into the
diameter, the distance from the top of the cylinder to the 400 mL beaker. Repeat the extraction with 2 additional
lower tip of the shaft being 2.54 em, and an immersion quantities, each of 25 ml., of waterR, decanting each time
depth of 5.00 em (No.2 spindle); if the scale reading is the supernatant through the filter into the 400 mL beaker.
greater than 90 per cent of the full scale, repeat the Wash the filter with 25 mL of hot water R, collecting this
measurement using a spindle similar to the No. 2 spindle but filtrate in the 400 mL beaker. Concentrate the combined
with a cylinder 1.27 cm in diameter and 0.16 cm high (No.3 filtrates to about 20 mL by gently boiling. If a precipitate
spindle). appears, add about 0.1 mL of n;m'c acidR, heat to boiling
For type IC, use a No.3 spindle; if the scale reading is and allow to cool to room temperature. Filter the
greater than 90 per cent of the full scale. repeat the concentrated extracts through a rapid-flow filter paper into a
measurement using a spindle with a cylindrical shaft 0.32 cm 50 mL volumetric flask. Transfer the remaining contents of
in diameter and an immersion depth of 4.05 cm (No.4 the 400 mL beaker through the filter paper and into the Oask
spindle). with water R. Dilute this solution to 50.0 mL with water R.
For types IB and lIA, use a No.2 spindle. Reference solutions Prepare the reference solutions using lead
standard solution (10 ppm Ph) R, diluted as necessary with
Limits:
water R.
. Source Lead hollow-cathode lamp.
Viscosity (mPa-s)
Typ. Wavelength 217 nm.
min. mex,
Atomisation detnce Oxidising air-acetylene flame.
lA 225 600 Loss on drying (2.2.32)
18 Maximum 8.0 per cent, determined on 1.000 g by drying in
1'0 4'0
an oven at 110 "C,
IC 800 2200
IlA 100 300 TAMC: acceptance criterion 103 CFU/g (2.6.12).
TYMC: acceptance criterion 10' CFUlg (2.6.12).
Acid demand Absence of Escherichia coli(2.6.13).
Weigh a quantity of the substance to be examined equivalent
ASSAY
to 5.00 g of the dried substance and disperse in 500 mL of
Alumlnlum
water R using a suitable blender fitted with alL jar. With
Atomic absorption spectrometry (2.2.23, Methad 1).
constant mixing, add 3.0 mL portions of 0.1 M hydrochloric
acid at 5 S, 65 s, 125 s, 185 s, 245 s, 305 s, 365 s, 425 s, Test salution In a platinum crucible mix 0.200 g with 1.0 g
485 s, 545 s, 605 s, 665 s and 725 s and add a 1.0 mL of anhydrous lithium metaborate R. Heat slowly at first and
portion at 785 s. The pH (2.2.3) determined at 840 s is not ignite at 1000-1200 °C for 15 min. Allow to cool, place the
greater than 4.0. crucible in a 100 mL beaker containing 25 mL of a
40 per cent VIV solution of dilute mtnc acidR and add
Arsenic (2.4.2, Me/hodA) 50 mL of a 40 per cent VIV solution of dilute nitric acid R,
Maximum 3 ppm. filling and submerging the crucible. Place a
Transfer 16.6 g to a 250 mL beaker containing 100 mL of polytetrafluoroethylene-coated magnetic stirring bar in the
dl1uc.e hydrochloric acid R. Mix, cover with a watch glass and crucible and stir gently with a magnetic stirrer until
boil gently, with occasional stirring, for 15 min. Allow the dissolution is complete. Transfer the solution to a 200 mL
insoluble matter to settle and decant the supernatant through volumetric flask, wash the beaker, crucible and magnetic
a rapid-flow filter paper into a 250 mL volumetric flask, stirrer bar with water R, collecting the washings in the
retaining as much sediment as possible in the beaker. To the volumetric flask, and dilute to 200.0 mL with water R
residue in the beaker add 25 mL of hot duute hydrochlonc (solution A). To 20.0 mL of solution A add 20 mL of a
acidR, stir, heat to boiling, allow the insoluble matter to 10 gIL solution of sodium chloride R and dilute to 100.0 mL
settle and decant the supernatant through the filter into the with water R.
volumetric flask. Repeat the extraction wilh 4 additional Reference solutions Dissolve, with gentle heating, 1.000 g of
quantities, each of 25 mL, of hot dilute hydrochloric acidR, aluminium R in a mixture of 10 mL of hydrochloric acid Rand
decanting each supernatant through the filter into the 10 mL of waterR. Allow to cool, then dilute to 1000.0 mL
volumetric flask. At the last extraction, transfer as much of with water R (1 mg of aluminium per millilitre). Into 4
the insoluble matter as possible onto the filter. Allow the identical volumetric flasks, each containing 0.20 g of sodium
combined filtrates to cool to room temperature and dilute to chloride R, introduce 1.0 mL, 2.0 mL, 3.0 mL and 4.0 mL of
250.0 mL with dilute hydrochloric acidR. Dilute 5.0 mL of this solution respectively, and dilute to 100.0 mL with
this solution to 25.0 mL with duute hydrochloric acidR. water R.
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1-126 Aluminium Phosphate 2022
Blank solutio" Dissolve 0.20 g of sodium chloride R In

water R and dilute to 100.0 mL with the same solvent.
Dried Aluminium Phosphate
Source Aluminium hollow-cathode lamp. (Aluminium Phosphate, Hydrated, Ph. Eur.
Wavelength 309 nm. monograph 1598)
Atomisation device Acetylene-nitrous oxide flame. A1PO..,xH,O 122.0
Magnesium (anhydrous substance)
Atomic absorption spectrometry (2.2.23, Method1).
Action and use
Testsolution Dilute 25.0 mL of solution A, prepared in the
Antacid.
assayfor aluminium, to 50.0 mL with waler R. To 5.0 mL of
this solution add 20.0 mL of lanthanum chloride solUlian R Phfll _
and dilure to 100.0 mL with water R.
DEFINITION
Reference solutions Place 1.000 g of magnesium R in a Content
250 mL beaker containing 20 mL of water R and carefully 94.0 per cent to 102.0 per cent of AlPO. (M, 122.0) (ignited
add 20 mL of hydrochloric acidR, warming if necessary '0 substance).
dissolve. Transfer the solution to a volumetric flask and
dilute to 1000.0 mL with waterR (I mg of magnesium per It contains a variable quantity of water.
'0
millilitre). Dilute 5.0 mL of this solution 500.0 mL with CHARACTERS
water R. Into 4 identical volumetric flasks) introduce 5.0 mL, Appearance
10.0 mL, 15.0 mL and 20.0 mL of the solution respectively. White or almost white powder.
To each flask add 20.0 mL of lanthanum chloride solution R Solubility
and dilute '0
100.0 mL with waterR. Veryslightly soluble in water, practically insoluble in ethanol
Blank solution Dilute 20 mL of lanthanum chloride solution R (96 per cent). It dissolves in dilute solutions of mineral acids
'0 100.0 mL with waterR. and alkali hydsoxides.
Source Magnesium hollow-cathode lamp. IDENTIFICATION
Wavelength 285 urn. A. Solution S (see Tests) gives reaction (b) of phosphates
Awmisation device Air-acetylene flame. (2.3.1).
LABELLING B. Solution S gives the reaction of aluminium (2.3.1).
The label states the ratio of aluminium content to TESTS
rnagnesiwn content) the viscosity and the corresponding type Solution S
(see Definition). Dissolve 2.00 g in dilul< hydrochlon"c acidR and dilute to
FUNCTIONALITY-RELATED CHARACTERISTICS 100 mL with the same acid.
This seaion provides information on characteristics that are Appearance of solution
recognised as being relevant control parameters for one or more Solution S is clear (2.2.1) and colourless (2.2.2, Method /1).
functions of thesubstance when usedas an excipient (see chapter pH (2.2.3)
5.15). Someof the characteristics described in the Funaionality-
related characunstia section may also be present in the mandatary
5.5 '0
7.2
part of the monograph since they alsorepresent mandatory quality Shake 4.0 g with carbon dioxide-free waterR and dilute '0
cntetia. In such cases, a cross-reference to the tests described in the
mandatory part is induded in the Functionalir:y-related Chlorides (2.4.4)
characteristics section. Control of the cbaraaoistics can contribute Maximum 1.3 per cent.
U! the quality of a medicinal product by improving the consistency Dissolve 50.0 mg in 10 mL of dilute niuic add R and dilute
of the manujactuting process and the performance of the medicinal to 200 mL with waterR.
product dun'ng use. Where control methods are cited, they are Soluble phosphates
recognised as being suitable for thepurpose, but other methods can Maximum 1.0 per cent, calculated as P04 3 -.
Test solution Stir 5.0 g with 150 mL of waterR for 2 h.
reported, the conlro1 method must be indicated.
Filter and wash the filter with 50 mL of waterR. Combine
Thefollowing characteristic may be relevant for aluminium the filtrate and the washings and dilute'0 250.0 mL with
magnesium silicate used as viscosity-increasing agent and
stabiliser.
'0
waterR. Dilute 10.0 mL of this solution 100.0 mL with
water R.
Viscosity Reference solution (a) Dissolve 28.6 mg of potassium
(see Tests). dihydrogen phosphate R in water Rand dilute to 100.0 mL
__ ~ PhEll with the same solvent. Dilute 10.0 mL of the solution to
Reference salution (b) Dilute 1.0 mL of reference solution (a)
'0 5.0 mL with waterR.
Rsference solution (c) Dilute 3.0 mL of reference solution (a)
'05.0 mL with waterR.
Treat each solution as follows. To 5.0 mL add 4 mL of dilute
sulfuric acid R, 1 mL of ammonium molybdate solution R, 5 mL
of water Rand 2 mL of a solution containing 0.10 g of
4-methylaminophenol sulfate R, 0.5 g of anhydrous sodium
sulfite Rand 20.0 g of sodium metabisulfite R in 100 mL of
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2022 Aluminium Phosphate Gel 1-127
water R. Shake and allow to stand for 15 min. Dilute to TESTS

25.0 mL with water R and allow to stand for a further SoJution S
15 min. Measure the absorbance (2.2.25) at 730 run. Dissolve 2.00 g in dilure hydrochlo'ic acid R and dilute to
Calculate the content of soluble phosphates from a 100 mL with the same acid.
calibration curve prepared using reference solutions (a), pH (2.2.3)
(b) and (c) after treatment. 6.0 to 8.0.
Sulfates (2.4.13) Peroxides
Maximum 0.6 per cent. Maximum 150 ppm, expressed as hydrogen peroxide.
Dilute 8 mL of solution S to 100 mL with distilled warer R. Testsolution Dissolve with heating 1.0 g of the substance to
Arsenic (2.4.2) be examined in 5 mL of dilute hydrodrloric acidR, then add
Maximum I ppm. 5 mL of water Rand 2 mL of divanadium pentoxide solution in
1.0 g complies with limit test A. sulfuric acidR.
Loss on ignidon Reference solution Dilute 1.0 mL of dilute hydrogen peroxide
10.0 per cent to 20.0 per cent, determined on 1.000 g at solution R to 200.0 mL with warer R. To I mL of this
800 ± 50°C. solution add 9 mL of warer R and 2 mL of divanadium
pentoxide solution in su/jurk acidR.
Neutralislng capacity
Add 0.50 g to 30 mL of 0.1 M hydrodrlonc acid previously The test solution is not more intensely coloured than the
heated to 37 °C and maintain at this temperature for 15 min reference solution.
while stirring. The pH (2.2.3) of the mixture after 15 min at Chlorides (2.4.4)
37 °C is 2.0 to 2.5. Maximum 500 ppm.
ASSAY Dissolve 1.3 g in 5 rnL of dilute niuic acid R and dilute to
Dissolve 0.400 g in 10 mL of dilute hydrochloric acid Rand 200 mL with warer R.
dilute to 100.0 mL with warer R. To 10.0 mL of the Soluble phosphates
solution, add 10.0 mL of 0.1 M sodium edetate and 30 mL of Maximum 0.5 per cent, expressed as PO".
a mixtureof equal volumes of ammonium aatale solution R Testsolution Centrifuge 10.0 g until a clear supernatant is
and diluteacetic acidR. Boil for 3 min, then cool. Add 25 mL obtained. To 2.00 mL of the supernatant add 20.0 mL of a
of ethanol (96 per cent) R and I mL of a fteshly prepared 10.3 gIL solution of hydrodr/oric acid R and dilute to
0.25 gIL solution of dithizone R in ethanol (96 per cent) R. 100.0 mL with warer R. To 10.0 mL of this solution add
Titrate the excess of sodium edetate with 0.1 M zin« sulfau 10.0 mL ofnitro-molybdovanadic reagent R and dilute to
until the colour changes' to pink. 50.0 mL with warer R. Allow to stand protected from light
I mL of 0.1 M sodium <delate is equivalent to 12.20 mg of for 15 min.
AlPO•. Reference solution Add 10.0 mL of nitro-molybdovanadic
STORAGE reagent R to 10.0 mL of a 143 mgIL solution of potassium
In an airtight container. dihydrogen phosphate R and dilute to 50.0 mL with warer R.
______________ ~ I'I>f" Allow to stand protected from light for 15 min.
Measure the absorbances (2.2.25) of the 2 solutions at
400 nm. The absorbance of the test solution is not greater
than that of the reference solution.
Aluminium Phosphate Gel Sulfates (2.4.13)

(Ph. Bur. monograph 2166) Dilute 25 mL of solution S to 100 mL with distiOed warer R.
Acdon and use Soluble aluminium
Antacid; vaccine adjuvant. Maximum 50 ppm.
POE" _ To 16.0 g add 50 mL of warer R. Heat to boiling for 5 min.
Cool and centrifuge. Separate the supernatant. Wash the
DEFINITION residue with 20 mL of water R and centrifuge. Separate the
Hydrated AlPO. in gel form, supernatant and add to the first supernatant. To the
Content combined supernatants add 5 mL of hydrochloric. acid Rand
19.0 per cent to 21.0 per cent of AlP04 . 20 mL of warer R. Introduce all of this solution into a
500 mL conical flask and carry out the complexometric
CHARACTERS titration of aluminium (2.5.11) using 0.01 M sodium <delate.
Appearance Arsenic (2.4.2, MethodA)
Gel. Maximum 1 ppm) determined on 1.0 g.
Solubility
Acid neutralising capacity
Practically insoluble in water, in ethanol (96 per cent) and in
Add 2.0 g to 30 mL of 0.1 M hydrochloric acid heated to
methylene chloride. It dissolves in dilute solutions of mineral
37°C and maintain at 37 °C. while shaking. Determine the
acids. pH after 15 min. The pH (2.2.3) of the mixture is 2.0 to 2.5.
IDENTIFICATION Residue on Ignition
A. Solution S (see Tests) gives reaction (b) of phosphates 19.0 per cent to 23.0 per cent.
(2.3.1).
Heat 0.500 g at 50°C for 5 hours) then ignite at
B. Solution S gives the reaction of aluminium (2.3.1). 500 ± 50°C until constant mass.
C. It complies with the assay.
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1-128 Aluminium Powder 2022
Microbial contamination Iron

TA.t\1C: acceptance criterion 10' CFU/g (2.6.12). Dissolve 10 mg in 20 mL of 2M hydrochloric acid and dilute
TYMC: acceptance criterion 10' CFU/g (2.6.12). to 100 mL with water. 10 mL of the resulting solution
Absence of bile-tolerant gram-negative bacteria (2.6.13). complies with the limit test/or iron, Appendix vn
(1.0%).
Absence of Escherichia coli(2.6.13). Lead
Use two solutions prepared in the following mariner.
ASSAY For solution (1) boil 0.40 g with 20 mL of 2M hydrochlori:
Dissolve with heating 0.300 g in 5 mL of dllwe hydrtXhloric acid and 10 mL of water until effervescence ceases) add
acid R. Add 45 mL of warer R, 10.0 mL of 0.1 M sodium 0.5 mL of nitric add) boil for 30 seconds and cool; add 2 g of
edetate and 30 mL of a mixture of equal volumes of ammonium chloride and 2 g of ammonium thiocyanate, extract
ammonium acetate solution Rand d11ute acetic acid R. Heat to with three 10 mL quantities of a mixture of equal volumes of
boiling and maintain boiling for 3 min. Cool, then add amyl akohol and ether, discard the extracts and add 2 g of
25 mL of ethanol (96 per cent) R. Titrate with 0.1 M zinc cime acid. For solution (2) dissolve 2 g of citric acid in 10 mL
sulfare, determining the end-point potentiometricaUy (2.2.20). of 2M hydrtXhloric acid and add 4 mL of leadstandard solurion
1 mL of 0.1 M zinc sulfate is equivalent to 12.2 mg of AlPO... (10 ppm Ph). Make solutions (1) and (2) alkaline with
5M ammonia and to each add 1 mL of pouusium cyanide
STORAGE
solution PbT. The solutions should be not more than faintly
opalescent. If the colours of the solutions differ, equalise by
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E"
the addition of about 0.2 mL of a highly diluted solution of
burnt sugar or other non-reactive substance. Dilute each
solution to 50 rnL with water, add 0.1 mL of a 10% wlv
solution of sodium sulfide to each and mix thoroughly.
Aluminium Powder The colour produced in solution (1) is not more intense than
that produced in solution (2), when viewed against a white
AI 26.98 7429-9/)-5
background (100 ppm).
Action and use Other metals
Topical protective. Dissolve 2 g in 40 mL of 2M hydrtXhloric acid. Dilute 20 mL
Preparation of the solution to 100 mL with water, make alkaline to litmus
Compound Aluminium Paste paper by the addition of 5M ammonia, boil and filter.
Evaporate the filtrate to dryness, add 0.05 mL of sulfuric acid
DEFINITION and ignite. The residue weighs not more than 2 mg.
Aluminium Powder consists mainly of metallic aluminium in Lubricant
the form of very small flakes, usually with an appreciable To 2 g add 100 mL or hot water) cover and add, drop wise,
proportion of aluminium oxide; it is lubricated with stearic sufficient of a mixture of equal volumes of hydrochlorit acid
acid to prevent oxidation. It contains not less than 86.0% of and water to dissolve the metal almost completely. Heat to
AI, calculated with reference to the substance freed from complete dissolution, cool, filter through a hardened filter
lubricant and volatile matter. paper and wash the vessel and filter paper thoroughly with
CHARACTERISTICS water; dry both the vessel and paper at room temperature.
A silvery grey powder. Extract the paper with thsee 100-mL quantities of boiling,
freshly distilled acetone, using the original vessel to contain
Practically insoluble in water and in ethanol (96%).
the solvent and then wash the paper wilh five 10-mL
It dissolves in dilute acids and in aqueous solutions of alkali
quantities of freshly distilled acetone. Evaporate the combined
hydroxides) with the evolution of hydrogen.
filtrate and washings to dryness using a rotary evaporator.
IDENTIFICATION The residue, after drying at 105 0 for 30 minutes and allowing
A solution in 2M hydrochloric add yields the reaction to cool, weighs 10 to 60 mg.
characteristic of aluminium salts) Appendix VI. When the basin containing the residue is floated in a beaker
TESTS of water suitably stirred and heated, the residue melts
Surface-covering power between 40 0 and 60°. The residue is almost completely
Not less than 4000 cm' per g when determined by the soluble, with effervescence, in hot dilute sodium carbonate
following method. Fill with water a shallow trough measuring solution.
approximately 60 cm x 12 cm x 1.5 ern, fitted with a Volatile matter
movable partition so constructed that it is a sliding fit and When heated to constant weight at 105'"', loses not more than
can be used to divide the trough into two rectangular areas. 0.5% of its weight. Use 1 g.
Place the movable partition near one end and sprinkle 50 mg
ASSAY
of the substance being examined on the surface of the liquid
Transfer 0.2 g, previously freed from lubricant by successive
confined in the smaller area. Using a glass rod, spread the
washing with acetone and drying) to a three-necked 500 mL
powder evenly over the liquid surface until an unbroken film
flask fitted with a 150 mL dropping funnel, an inlet tube
covers the entire surface. Move the partition so as to increase
connected to a cylinder of carbon dioxide and an outlet tube
the area confined and again spread the powder to cover the
dipping into a water trap. Add 60 mL of warer and disperse
increased surface. Continue this process and determine the
the substance being examined; replace the air by carbon
maximum unbroken surface area obtained. The surface-
dioxide and add 100 mL of a solution containing 56 g of
covering power is the area covered per g of the powder at the
ammonium iron(m) sulfate and 7.5 mL of sulfuric acid in water.
breaking point of the film,
While maintaining an atmosphere of carbon dioxide in the
flask, heat to boiling) boil for 5 minutes after the sample has
dissolved, cool rapidly to 20 0 and dilute to 250 mL with
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2022 Aluminium Sodium Silicate 1-129
water. To 50 mL add 15 mL of orthophosphoric acid and filter into the beaker. Repeat the extraction with 2 additional
titrate with O.02M potassium permanganate VS. Each mL of quantities, each of 25 ml., of hot water R) decanting each
O.02M potassium permanganate VS is equivalent to 0.8994 mg supernatant through me filter into the beaker. Wash the filter
ofAl. with 25 mL of hot water R, collecting the filtrate in the
beaker. Concentrate the combined filtrates by gently boiling
to about 15 mL. Add about 0.05 mL of heavy metal-free ni,ric
add R, heat to boiling and allowto cool to room
Aluminium Sodium Silicate temperature. Filter the concentrated extracts through a rapid-
flow filter paperinto a 25 mL volwnetric flask. Transfer the
(Ph. Eur. monograph 1676) remaining contents of the beaker through the filter paper and
Pl>E<r _ into the volumetric flask with water R and dilute to 25.0 mL
with the same solvent.
DEFINITION Reference solutions Into 4 separate 100 mL volumetric flasks,
Silicicacid aluminium sodium salt of synthetic origin. introduce respectively 3.0 rnL, 5.0 rnL, 10.0 mL and
Content 15.0 mL of lead standard solution (10 ppm Pb) R, add
- aluminium (AI; Me 26.98): 2.7 per cent to 7.9 per cent 0.20 mL of heavy metal-free min'c acidR and dilute to
(driedsubstance); 100.0 mL with water R.
- sodium (Na; M, 22.99): 3.7 per cent to 6.3 per cent (dried Source Lead bollow-cathode lamp.
substance). Wavelength 217.0 om.
CHARACTERS Atomisauon device Air-acetylene flame.
Appearance Loss on drying (2.2.32)
White or almost white) fine, light) amorphous powder. Maximum 8.0 per cent, determined on 1.000 g by drying in
Solubility an oven at 105 QC for 4 h.
Practically insoluble in water and in organic solvents. Loss on ignition
IDENTIFICATION 5.0 per cent to 11.0 per cent (dried substance), determined
A. Transfer 1.0 g to a 100 mL beaker and add 10 mL of on 1.000 g by ignition in a platinum crucible to constant
dilute hydrochlork acid R. N1ix, cover with a watch glass and mass at 1000 ± 25°C.
boil for 15 min. Allow to cool to room temperature, mix and Microbial contamination
centrifuge the solution. 2 mL of the supernatant gives the TAMC: acceptance criterion 10' CFU/g (2.6.12).
reaction of aluminium (Z.3.1). TYMC: acceptance criterion 102 CFU/g (2.6.12).
B. 2 mL of me supernatant obtained in identification test A Absence of Escherichia coli (2.6.11).
gives reaction (a) of sodium (2.3.1).
ASSAY
C. 0.2 g gives the reaction of silicates (2.3.1).
Aluminium
TESTS Atomic absorption spectrometry (2.2.23, Method l).
pH (2.2.3) Acid mixture Add 50 mL of "itric acid R to 500 mL of
9.5 to 11.5. water R. Dissolve in this solution 17 g of tanarK. acid Rand
Disperse 5.0 g in 100 mL of carbon dioxide-free water R. dilute to 1000 mL with water R.
Arsenic (2.4.2, Method A) Blank solution Dissolve 1.4 g of anhydrous lithium
Maximum 3 ppm. metaborate R in 60 mL of the acid mixture and dilute to
Transfer 8.3 g to a 250 mL beaker containing 50 mL of 200 mL with water R.
dilute hydrochloric acid R. Mix, cover with a watch glass and Test solution In a platinum crucible mix 0.200 g with 1.4 g
boil gently, with occasional stirring, for 15 min. Centrifuge, of anhydrous lithium metaburate R. Heat slowly at first and
and decant the supernatant through a rapid-flow filterpaper ignite at 1100 ± 25°C for 15 min. Cool) chen place the
into a 250 mL volumetric flask. To the residue in the beaker, crucible in a 100 mL beaker containing 60 mL of the acid
add 25 mL of bot dilute hydrochlotic acidR, stir, centrifuge, mixture. Place a polytetrafluoroethyJene-coated magnetic
and decant the supernatant through the same filter into the stirring bar in the crucible and stirgently with a magnetic
volwnetric flask. Repeat the extraction with 3 additional stirrer for 16 h. Transfer the contents of the crucible into a
quantities, each of 25 rnL, of hot dilute hydrochloric acid R, 200 mL volumetric flask. Wash the crucible, the magnetic
filtering each supernatant through this filter into the stirring bar and the beaker with water R and dilute to
volumetric flask. Allow the combined fila-ares to cool to room 200.0 mL with the same solvent (solution A). To 10.0 mL of
temperature and dilute to 250.0 mL with dilute hydrochloric this solution) add 1.0 mL of lanthanum chloride solution Rand
acid R. Dilute 10.0 mL of the solution to 25.0 mL with dilute to 50.0 mL with water R.
water R. Reference solutwns Into 5 separate 50 mL volumetric flasks,
Lead inttoduce respectively 1.0 101., 2.5 rnL, 5.0 rnL, 7.5 mL and
Maximum 5 ppm. 10.0 mL of aluminium standardsolution (100 ppm AD R, add
Atomic absorption spectrometry (2.2.23, Method l). I mL of lanthanum chloride solution R and 10 mL of the blank
solution, and dilute to 50.0 mL with water R.
Test solution Transfer 5.0 g to a 250 mL beakercontaining
50 rnL of dilute hydrochloric acid R. Mix, cover with a watch Source Aluminium hollow-cathode lamp.
glass andboil for 15 min. Allow to cool to room Wavelength 309.3 om.
temperature. Centrifuge, and decant the supernatant through Atomisauon deoice Acetylene-nitrous oxide flame.
a rapid-flow filter paper into a 250 rnL beaker. To the Sodium
insoluble matter add 25 mL of hot water R. Stir vigorously, Atomicemissionspectrometry (2.2.22, lvlethod 1).
centrifuge, anddecant the supernatant through the same
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1-130 Aluminium Stearate 2022
Testsolution To 2.0 mL of solution A, prepared in the assay distilled waterR (solution S). Evaporate the ether layer to
of aluminium, add 1 mL of a 12.5 gIL solution of caesium dryness and dry the residue at 100-105 "C. Keep the residue
chloride R and dilute to 20.0 mL with walei' R. for identification tests A and B.
Reference solutions Into 5 separate 200 mL volumetric flasks, Acidity or alkalinity
each containing 10 mL of a 12.5 gIL solution of caesium To 1.0 g add 20 mL of carbon dioxide-free walei' R and boil
chloride R, introduce respectively 1.0 mL, 2.0 mL, 4.0 mL, for 1 min with continuous shaking. Cool and filter.
6.0 mL and 10.0 mL of sodium standard solution To 10 mL of the Iiltrate add 0.05 mL of bromorhymol blue
(ZOO ppm Na) R and dilute to 200.0 mL with walel'R. solution R4. Not more than 0.05 mL of 0.1 M hydrochloric
Wavelengrh 589.0 om. acid or 0.1 M sodium hydroxide is required to change the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I colourof the indicator.
Chlorides (2.4.4)
Dilute 0.5 mL of solution S to 15 mL with water R.
Aluminium Stearate ***
** ** Sulfates (2.4.13)
Maximwn 0.5 per cent.
(ph. Eur. monograph 1663) *****
Dilute 0.3 mL of solution S to 15 mL with distilled walei' R.
PhE" _
Cadmium
DEFINITION Maximum 3 ppm.
Aluminium salts of a mixture of solid organic acids consisting Atomic absorption spectrometry (2.1.23, Method II).
mainly of variable proportions of aluminium stearate and an
For thepreparation of aqueous solutions andfor the rinsing of
aluminium palmitate. The organic acids are obtained from glaJSWare be/ore use, employ waterrhathas been passed through a
sources of vegetable or animalorigin. strong-acid, strong-base, mixed-bed ion-exchange resin before use.
Content Select all reagents to have as low a content of cadmium, leadand
- aluminium (Al; A r 26.98): 3.0 per cent to 9.0 per cent nicJuI as practicable and store all reagent solutions in containers of
(dried substance); borosilicate glass. Clean glassware be/ore use by soaking in a
- steaM"c acid in thefatty add fraaion: minimum wann 773 gIL solution of nim"c acid R for 30 min and by rinsing
40.0 per cent; with deionised water.
- sum of stearic acid and palmitic acid it' the fatly add fraction: Bklnk solution Dilute 25 mL of cadmium- and lead-free nitric
minimum 90.0 per cent. acidR to 100.0 mL with walel'R.
CHARACTERS Modifier solution Dissolve 20 g of ammonium dihydrogen
Appearance phosphate Rand 1 g of magnesium nitrate R in waterR and
White or almostwhite, very fine, light powder. dilute to 100 mL with the same solvent. Alternatively, use an
Solubility appropriate matrix modifier as recommended by the graphite
Practically insoluble in water and in anhydrous ethanol. furnace atomicabsorption (GFAA) spectrometer
manufacturer.
IDENTIFICATION Test solution Place 0.100 g of the substance to be examined
First identification: C, D. in a polytetmfluoroethylene digestion bomb and add 2.5 mL
Second identification: A, B, D. of cadmium- and lead-free nitric acid R. Close and seal the
A. Freezing point (2.2.11f): minimum 53 "C, determined on bomb according to the manufacturer's operating instructions.
the residue obtained in the preparation of solution S When using a digestion btmlb.J be thoroughly familiar with the
(see Tests). safct)! and operating instructions. Carefully follow the bomb
B. Acid value (2.5.1): 195 to 210. manu/aemreY's instructions regarding care and maintenance 0/
Dissolve0.200 g of the residue obtained in the preparation of
these digestion bombs. Do not use metal-jacketed bombs or liners
thaI hatJe been used with hydrochloric acid due to contamination
solutionS in 25 mL of the prescribed mixture of solvents,
from corrosion of the metaljacket by hydrochloric acid. Heat the
C. Examine the chromatograms obtained in the assay of bomb in an oven at 170 QC for 3 h. Cool the bomb slowlyin
stearic acid and palmitic acid. air to room temperature according to the bomb
Results The 2 principal peaks in the chromatogram obtained manufacturer's instructions. Place the bomb in a fume
with the test solution are similar in retention time (0 the cupboard and open carefully as corrosive gases maybe
2 principal peaks in the chromatogram obtained with the expelled. Dissolve the residue in waterR and dilute to
reference solution. 10.0 mL with the same solvent.
D. 1 mL of solutionS gives the reaction of aluminium Reference solution Prepare a solutioncontaining
(2.3.1). The addition of 0.5 mL of dilute hydrochloric acidR 0.00165 ~!¥mL of cadmium nitrate ",,"hydrate R in the blank
described in the general method is omitted. solution (equivalent to 0.006 ~!¥mL of Cd).
TESTS Dilute 1.0 mL of the test solution to 10.0 mL with the blank
Solution S solution. Prepare mixtures of this solution, the reference
To 5.0 g add 50 mL of peroxide-free ether R, 20 mL of dilute solution and the blank solution in the following proportions:
nitric acid Rand 20 mL of distilled walei' R and heat gently (1.0:0:1.0 VfVll'), (1.0:0.25:0.75 VIVII'),
under a reflux condenser until dissolution is complete. Allow (1.0:0.5:0.5 VIVII'), (1.0:0.75:0.25 VIVII'). To each mixture
to cool. In a separating funnel, separate the aqueous layer add 50 ~L of the modifier solutionand mix. These solutions
and shake the ether layer with 2 quantities, each of 4 mL, of contain respectively 0 ~g, 0.0015 pg, 0.0030 ~g and
distilled water R. Combine the aqueous layers, wash with 0.0045 Jig of cadmium per millilitre from the reference
15 mL of peroxide-free ether R and dilute to 50.0 mL with
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2022 Aluminium Stearate 1-131
solution. Keep the remaining test solution for use in the test For the preparation of alJ aqueous solutions andfor the rinsing of
for lead and nickel. glassware before use, employ water that has been passed through a
Source Cadmium hollow-cathode lamp. strong-acid, strong-base, mixed-bed ion-exchange resin before use.
Wavelength 228.8 nm. Select aU reagents to have as low a content of cadmium, leadand
nickel as practicable and store all reagent solutions in containers of
Atomisation device Furnace. borosilicate glass. Clean glassware before use by soaking in a
Pkuform Pyrolyrically coated with integrated tube. warm 773 gIL sohuion of nitric add R for 30 min and by n'ming
Operating conditions Use the temperature programme with deionised water.
recommended forcadmium by the GFAA manufacturer. Blank solution Use the solutiondescribed in the test for
An example of temperature parameters for GFAA analysis of cadmium.
cadmium is shown below. Modifier solution Dissolve 20 g of ammonium dihydrogen
phosphate R in water R and dilute to 100 mL with the same
Stage Final temperature Ramp time Hold time
solvent. Alternatively, use an appropriate matrix modifier as
rCJ (,j (,j
recommended by the GFAA spectrometer manufacturer.
Drying 110 10 20
Ashing 600 10 30
Testsolution Use the solution described in the test for
Atomisation 1800 0 5
cadmium.
Reference solution Prepare a solution of 0.050 ~glrnL ofNi
by suitable dilutions of a 0.2477 ~glmL solution of nickel
Lead
nitratehexahydrate R with the blank solution.
Maximum 10 ppm.
Prepare mixtures of the test solution, the reference solution
Atomic absorption spectrometry (2.2.23, Method II).
and the blank solutionin the following proportions:
For thepreparau"Q1l of all aqueous solutions andfor the rinsing of (1.0:0: 1.0 VIVIV), (1.0:0.5:0.5 VIVIV), (1.0: 1.0:0 VIVIV).
glassware before useJ employ waterthat has bun passed through a To each mixture add SO ~L of the modifier solutionand mix.
strong-acid, strong-base, mixed-bed ion-exchange resin brim use. These solutions contain respectively 0 ug, 0.0125 ~g and
Select all reagents lO have as low a content of cadmium, lead and 0.025 ~lg of nickel per millilitre from the reference solution.
nkkel as praaicoble and stow aU rMgent solutions in containers of
Source Nickel hollow-cathode lamp.
borosilicate glass, Clean glassware before use by soaking in a
wann 773 gIL solution of nioic acid RfOT 30 min and by rinsing Wavelength 232.0 nm.
wuh deionised water. Atomisation device Furnace.
Blank solution Use the solurion described in the test for Platform Pyrolytically coated with integrated tube.
cadmium. . Operating conditions Use the temperature programme
Modifier solution Use the solution described in the test for recommended for nickel by the GFM manufacturer.
cadmium. An example of temperature parameters for GFAA analysis of
Test solution Use the solution described in the test for nickel is shown below.
cadmium.
Stage Finallemperature Rampdme Hold time
Reference solution Prepare a solution of 0.1 00 ~glrnL of Pb rCJ (,j (,)
by suitable dilutions of lead standardsolution (100 ppm Pb) R
Drying 110 10 20
with the blank solution.
Ashing 1000 20 30
Prepare mixtures of the test solution, the reference solution Atomisation 2300 0 5
and the blank solution in the following proportions:
(1.0:0:1.0 VIVIV), (1.0:0.5:0.5 VIVIV), (1.0:1.0:0 VIVIV).
To each mixture add 50 JlL of the modifier solution and mix.
These solutions contain respectively 0 pg, 0.025 p.g and
0.05 p.g of lead per millilitre from the reference solution. an oven at 105 "C.
Source Lead hollow-cathode lamp. Microbial contaminadon
TAJ'AC: acceptance criterion 10 3 CFU/g (2.6.12).
TYMC: acceptance criterion 10 2 CFU/g (2.6.12).
Atomisation device Furnace.
Pkuform Pyrolyrically coated with integrated tube.
Absence of Salmonella (2.6.13).
Operating conditions Use the temperature programme
recommended forlead by the GFAA manufacturer. ASSAY
An example of temperature parameters for GFAA analysis of Aluminium
lead is shown below. To 0.250 g in a 250 rnL conical flask add 20 rnL of
methanol R and, slowly, 2 mL of sulfun'c addR. Heat the
Stage Final temperature Ramp time Hold time solutionfor 30 min under reflux on a water-bath, swirling
rCJ (,) (s) frequently. Allow to cool. Add 100 mL of waw R and adjust
Drying IJO 10 20 to about pH I by adding approximately 12 mL of diJul<
Ashing 450 10 30 sodium hydroxide solution R. Add 20.0 rnL of 0.1 M sodium
Atomisation 2000 0 5 edetate and adjust to between pH 5 and pH 6 by the addition
of sodium acetal< R. Add 70 mg of xy1enol orange triturate R
Nickel and titrate immediately and quickly with 0.1 M zinc sulfate
Maximum 5 ppm. until the colour changes from yellow to pinkish-violet.
Atomic absorption spectrometry (2.2.23, Method II). I mL of 0.1 M sodium edetate is equivalent to 2.698 mg of
AI.
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1-132 Aluminium Sulfate 2022
Stearic acid and palmitic acid CHARACTERS

Gas chromatography (2.2.28): use the normalisation Appearance
procedure. Colourless, lustrous crystals or crystalline masses.
Test solution In a conical flask fitted with a reflux condenser, Solubility
dissolve 0.100 g of the substance to be examined in 5 mL of Soluble in cold water, freely soluble in hot water, practically
boron tnjluoride-methanol solution R. Boil undera reflux insoluble in ethanol (96 per cent).
condenser fnr 10 min. Add 4 mL of heptane R through the
condenser and boil again undera reflux condenser for IDENTIFICATION
10 min. Allow to cool. Add 20 mL of saturated sadium A. Solution S (see Tests) gives reaction (a) of sulfates (2.3.1).
chloride solution R. Shake and allow the layers to separate. B. Solution S gives the reaction ofalurninium (2.3.1).
Dry the organic layer over 0.1 g of anhydrous sadium sulfate R TESTS
previously washed with heptane R. Dilute 1.0 mL of the Solution S
solution to 10.0 mL with heptane R. Dissolve 2.5 g in water R and dilute to 50 mL with the same
Reference solution Prepare the reference solution in the same solvent.
manner as the test solution using 50.0 mg of palmitic Appearance of solution
acidCRS and 50.0 mg of stearic acid CRS instead of the Solution S is not more opalescent than reference
substance to be examined.
suspension III (2.2.1) and is colourless (2.2.2, Me/had11).
Column:
pH (2.2.3)
2.5 to 4.0.
- size: I::: 30 m, 0 ::: 0.32 mm;
- stationary phase: macrogol 20 000 R (film thickness Dissolve 05 g in carbon dioxide-flu waterR and dilute to
0.5 um). 25 ml, with the same solvent.
Carrier gas helium for chromatagraphy R. Alkali and alkaline-earth metals
FI<>w rate 2.4 mllmin.
To 20 mL of solution S add 100 mL of warer R, heat and
Temperature:
add 0.1 mL of methyl red solution R. Add dilute ammonia Rl
Thn, Temperature until the colour of the indicator changes to yellow. Dilute to
(mln) ("C) 150 mL with water R, heat to boiling and filter. Evaporate
Column 0-2 70 75 mL of the filtrate to dryness on a water-bath and ignite.
2 - 3~ 70 240
-"-* The residue weighs a maximum of 2 mg.
36 - 41 2.0 Ammonium (2.4.1)
Inje<:tion port 220 Maximum 500 ppm.
Detector 260 Dilute 0.4 mL of solution S to 14 mL with water R.
Iron (2.4.9)
Detection Flame ionisation.
Maximum 100 ppm.
Injection I ~L.
Relative retention With reference to methyl stearate: methyl
Use 0.3 mL of thiog/ywUi< acidR in this test.
palmitate = about 0.9.
System suitability Reference solution: ASSAY
- resolution: minimum 5.0 between the peaks due to methyl Dissolve 0.500 gin 20 mL of water R. Carry out the
palmitate and methyl stearate; complexometric titration of aluminium (2.5.11).
- repeatability: maximum relative standard deviation of I mL of 0.1 1\1 sodium edetate is equivalent to 17.11 mg
3.0 per cent for the areas of the peaks due to methyl of A12(SO,),.
palmitate and methyl stearate after 6 Injections; maximum STORAGE
relative standard deviation of 1.0 percent for the ratio of
the areas of the peaks due to methyl palmitate to the areas
______ ~ PIIE<I
of the peaks due to methylstearate after 6 injections.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<I
Alverine Citrate
Aluminium Sulfate
Aluminium Sulphate
A1,(SO.),,xH20 342.1
(anhydrous substance)
Preparation
Aluminium Acetate Ear Drops
PIIE<I _ C,Jf,,NO, 473.6 5560-59-8
DEFlNITlON Action and use

Content Smooth muscle relaxant; antispasmodic.
51.0 per cent to 59.0 per cent of A1,(SO,h Preparation
It contains a variable quantity of water of crystallisation. Alverine Capsules
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2022 Alverine Citrate 1-133
""E<T _ Temperature:
DEFINITION
Thne Temperature
N-Ethyl-3-phenyl-N-(3-phenylpropyl)propan-l-amine (min) ('C)
dihydrogen 2-hydroxypropane-l,2,3-tricarboxylate.
Column 0-7 120
Content 7 - 13 120 --0 240
99.0 per cent to 101.0 per cent (dried substance). 13 - 21 240
CHARACfERS 21 - 24 240 ---> 290
Appearance 24 - 39 290
White or almost white, crystalline powder. Injection poet 290
Detector 290
Solublllty
Slightly soluble in water and in methylene chloride, sparingly
soluble in ethanol (96 per cent). Detection Flame ionisation.
Injection I ~L.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24). Identification of impurities Use the chromatogram obtained
wilh reference solution (c) to identify the peak due to
Comparison alven"ne citrate CRS. impurity C; use the chromatogram obtained with reference
TESTS solution (a) to identify the peak due to impurity D.
pH (2.2.3) Relat1've retention With reference to alverine (retention
3.5 to 4.5. time = about 18 min): impurity C = about O.5j
Dissolve 0.250 g in carbon dioxide-free water R and dilute to =
impurity D about 0.97.
50.0 mL with the same solvent. System suitability Reference solution (a):
Related substances - resolution: minimum 3.0 between the peaks due to
Gas chromatography (2.2.28): use the normalisation impurity D and alverine.
procedure. Use freshly prepared solutions. Limits:
Test solution Dissolve 0.250 g of the substance [0 be - impun"ties C, D: for each impurity, maximum
examined in water R and dilute to 20 mL with the same 0.15 per cent;
solvent. Add 2 mL of concentrated ammonia R and shake with - unspecified "mpurities: for each impurity, maximum
3 quantities, each of 15 mL, of melhy/ene chloride R. To the 0.10 per cent;
combinedlowerlayers add anhydrous sodium sulfate R, shake, - total: maximum 0.5 per cent;
filter, and evaporate the filtrate by suitable means at a - reporting threshold: 0.05 per cent (reference solution (b)).
temperature not exceeding 30°C. Take up the residue with Loss on drying (2.2.32)
methylene chloride R and dilute to 10.0 mL with the same Maximum 0.5 per cent, determined on 1.000 g by drying in
solvent. an oven at 80°C for 2 h.
Reference souaion (a) Dissolve 5 mg of a/verine Sulfuted ash (2.4.14)
impurity D CRS (impurity D citrate) in 5 mL of water R, add Maximum 0.1 per cent, determined on 1.0 g.
1 mL of concentrated ammonia R and shake with 3 quantities,
each of 5 mL, of methylene chloride R. To the combined lower ASSAY
layers add anhydrous sodium sulfate R, shake, filter, and Dissolve 0.375 g in 50 mL of anhydrous acetic acid R. Titrate
evaporate the filtrate by suitable means at a temperature not with O. J M perchlon"c acid, determining the end-point
exceeding 30 "C. Take up the residue with methylene potentiometrically (2.2.20).
chloride R, add 0.2 mL of the test solution and dilute to I mL of 0.1 M perchloric acid is equivalent to 47.36 mg
2.0 mL with methylene chloride R. of C26H3SN07'
Reference solution (b) Dilute 1"0 mL of the test solution to STORAGE
100.0 mL with melitylene chloride R. Dilute 1.0 mL of this Protected from light.
solution to 20.0 mL with methykne chloride R.
IMPURITIES
Reference solution (c) Dissolve 20 mg of alven'ue
Specified impurities C, D.
impurity C CRS in methylene chloride R and dilute to 20.0 mL
with the same solvent. Dilute 1.0 mL of the solution to Otherdetectable impurities (thefollowing substances would, if
10.0 mL with methylene chloride R. present at a sufficient level, be detected by Me or other of the tests
in the monograph. They arelimited by thegeneral acceptance
Column:
criterion for other/unspecified impurities and/or by thegeneral
- size: 1= 25 ill, 0 = 0.32 mm;
therefore not n«essary W identify these lmpun"ties for
- stationary phase: phenyl(5)methyl(95)polysiloxane R (film
demonstration of rompliance. See also 5.10. Control of impurities
thickness 0.45 urn).
in substances for pharmaceutical use) A, B, E.
Split ratio I: II.
A. l-chloro-3-phenylpropane,
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1-134 Amantadine Hydrochloride 2022
~OH
dried in vaalO at 60'C for I h, melts (2.2.14) at 147 "C to
151 'C.
C. Dissolve 0.2 g in I mL of 0.1 M hydroehlori< acid.
B. 3-phenylpropan-l-ol, Add I mL of a 500 gIL solution of sodium nitrite R. A white
precipitate is formed.
D. I mL of solution S (see Tests) gives reaction (a) of
chlorides (2.3.1).
TESTS
C. N-ethyl-3-phenylpropan-I -amine,
Solutlon S
25 mL with the Same solvent.
D. N-(3-cyclohexylpropyl)-N-ethyl-3-phenylpropan-I-amine, Solution S is clear (2.2.1) and not mOre Intensely coloured
man reference solution Y1 (2.2.2, J.Wethod II).
Acidity or a1ka1inlty
waterR. Add 0.1 mL of me/kyl red solution Rand 0.2 mL of
0.01 M sodium hydroxide. The solution is yellow. Add 0.4 mL
of 0.01 M hydrochloric acid. The solution is red.
Related substances
E. 3-phenyl-N,N-bis(3-phenylpropyl)propan-I -amine. Gas chromatography (2.2.21f).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEIT Internal standard solution Dissolve 0.500 g of adamantane R
in merhy1ene chloride R and dilute to 10.0 mL with the same
solvent.
Test sdution Weigh 0.5 g of the substance to be examined
Amantadine Hydrochloride ***
*** ***
into a centrifuge tube. Add 9 mL of methylene chlmide R and
10 mL of a 210 gIL solution of sodium hydroxide R. Shake for
(ph. Bur. monograph 0463) *** 10 min. Discard. the upperlayer. Dry the lowerlayer over
anhydrous sodium sulfate R. Filterand collect the filtrate in a
NH2
hY
V ,Hel
volumetric flask. Add 0.1 mL of the internal standard
solution and dilute to 10.0 mL with methylene chloride R.
Reference solution Weigh 5 mg of amantadine
hydrochloride CRS into a centrifuge tube. Add 9 mL of
187.7 665-66-7
methylene chloride Rand 10 mL of a 210 gIL solution of
Action and use sodium hydroxide R. Shake for 10 min. Discard the upper
Viral replication inhibitor (influenza A); dopamine receptor layer. Dry the lower layer over anhydrous sodium sulfate R.
agonist; treatment of influenza and Parkinson's disease. Filter and collect the filtrate in a volwnetric flask.
Add 1.0 mL of the internal standard solution and dilute to
Preparations 100.0 mL with me/kylene chloride R.
Amantadine Capsules
Column:
Amantadine Oral Solution - material: fused silica;
PhEIT _ - size: 1;;;; 30 m, 0 ;;;; 0.53 mm;
- stationary phase: base-deactivated phenyl(5)methy/(95)
DEFINITION polysiloxane R (film thickness I urn).
Tricyclo[3.3. I. I3"ldecan- I-amine hydrochloride. Carrier gas helium for chromatography R.
Content Flow rate 4 mllmin.
Split ratio I :50.
CHARACTERS Temperature:
Appearance
White or almost white, crystalline powder. Time Temperature
(mIn) CCl
Solubility
Freely soluble in water and in ethanol (96 per cent). Column 0·5 70
5·23 70 250
--->
It sublimeson heating.
23 - 40 250
IDENTIFICATION Injection port 220
First identification: A, D. Detector 300
Second identification: B. C, D.
A. Infrared absorption spectrophotometry (2.2.24). Detection Flame ionisation.
Comparison amantadine hydrochloride CRS. Injertion I ~L.
B. To 0.1 g add 1 mL of pyridine R, mix and add 0.1 mL of Relative retention Wil:h reference to amantadine (retention
acetic anhydride R. Heat to boiling for about 10 s. Pour the ume « about 14 min): internal standard e about 0.8.
hot solution into 10 mL of dilute hydrochlonc acid R, cool to
5 °C and filter. The precipitate, washed with water Rand
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2022 Ambroxol Hydrochloride 1-135
System suitability Referencesolution:

- resolution: minimum 5.0 between the peaks due to the
Ambroxol Hydrochloride
internal standard and amantadine. (ph. Bur. monograph 1489)
Limits:
- unspecified impurities: calculate the ratio CRt) of the areaof
the peak due [0 amantadine to the area of the peak due to
the internal standard from the chromatogram obtained HO
with the reference solution; from the chromatogram
obtained with the test solution, calculate the ratio of the
areaof any peak, apart from the principal peak and the
peak due to the internal standard, to the area of the peak
due to the internal standard: this ratio is not greaterthan 414.6 21828-92-4
R, (0. IO per cent);
- total: calculate the ratio (R2 ) of 3 times the areaof the Action and use
peak due to amantadine to the areaof the peak due to the Mucolytic expectorant.
internal standard from me chromatogram obtained with PIIE<B _
the reference solution; from me chromatogram obtained
with the test solution, calculate the ratio of the sum of the DEFINITION
areas of any peaks, apart from the principal peak and the trans-4-[(2-Amino-3,5-dibromobenzyl)aminojcyc!ohexanol
peak due to the internal standard, to the area of the peak hydrochloride.
due to the internal standard: this ratio is not greater than Content
R, (0.3 per cent); 99.0 per cent to 101.0 per cent (dried substance).
- disregard limit: calculate the ratio (R,) of 0.5 times the
area of the peak due to amantadine to die areaof the CHARACTERS
peak due to the internal standard from the chromatogram Appearance
obtained with the reference solution; from the White or yellowish, crystalline powder.
chromatogram obtained with die test solution, calculate Solubility
the ratio of the areaof any peak, apartfrom the principal Sparingly soluble in water, soluble in methanol, practically
peak and the peakdue to the internal standard, to the insoluble in methylene chloride.
area of the peak due to the internal standard: disregard IDENTIFICATION
any peak with a ratioless than R3 (0.05 per cent).
First identification: B, D.
Water (2.5.12) Second ideneficasion: A, C, D.
A. Ultraviolet and visible absorption spectrophotometry
Sulfated ash (2.4.14) (2.2.25).
Test solutian Dissolve 20.0 mg in dilute ,ulfuric acid Rl and
ASSAY dilute to 100.0 mL with the same acid. Dilute 2.0 mL of the
Dissolve 0.150 g in a mixture of 5.0 mL of 0.01 M solution ro 10.0 mL with dilute su/juri< acid Rl.
hydrochloric acid and 50 mL of erhanol (96 per ceno R. Carry Spearal range 200-350 run.
out a potentiometric titration (2.2.20), using 0.1 M sodium Absorption maxima At 245 om and 310 run.
hydroxide. Read the volume added between the 2 points of
inflexion. Absorbanu ratio A249'A310 = 3.2 to 3.4.
I mL of 0.1 M sodium hydroxide is equivalent to 18.77 mg of B. Infrared absorption spectrophotometry (2.2.24).
C,oH 18ClN. Comparison ambroxol hydrvchlcride CRS.
IMPURITIES C. Thin-layer chromatography (2.2.27).
Orher detectable impurities (rhe following substances would, if Testsolution Dissolve 50 mg of the substanceto be
present at a sufficient Ieud, be detected by one or otherof (he tests examined in methanol R and dilute to 5 mL with the same
in the monograph. They are limited by the general acaptance solvent.
criterion for other/unspecified impurities and/or by the general Reference solution Dissolve 50 mg of ambroxol
monograph Substances for pharmaceutical use (2014). It is hydrochloride CRS in methanol R and dilute to 5 mL with the
therefore not necessary to identify these impurities for same solvent.
demonstration of compliance. See also 5.1-0. Control 0/impumies Plate TLC silica gel Fm plate R.
in substances for pharmaceutical use) A, B. Mobile phase concentrated ammonia RJ propanol R, ethyl
0 acetate R, hexane R (1:10:20:70 VIVIV/V).
C25 Application 10 ~L.

Drying In air.
A. l-cWorotricyc!0[3.3.1.1"'jdecane,
Detection Examine in ultraviolet light at 254 run.
principal spot in me chromatogram obtainedwith the
reference solution.
B. N-(tricyc!0[3.3.1.1 "'jdec-I-yl)acetamide. D. Dissolve 25 mg in 2.5 mL of water R, mix with 1.0 mL of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<B dilute ammonia RJ and allow to stand for 5 min. Filter and
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1-136 Ambroxol Hydrochloride 2022
acidify the filtrate with dilute nitn"c add R. The filtrate gives Loss on drying (2.2.32)
reaction (a) of chlorides (2.3.1). Maximum 0.5 per cent, determined on 1.000 g by drying in
TESTS an oven at 105 "C.
Solution S Sulfated ash (2.4.14)
Dissolve 0.75 g in methanol R and dilute to 15 mL with the Maximum 0.1 per cent, determined on 1.0 g.
same solvent. ASSAY
Appearance of solution Dissolve 0.300 g in 70 mL of ethanol (96 per anO R and add
Solution S is clear (2.2.1) and not more intensely coloured 5 mL of 0.01 M hydrochloric acid. Carry out a potentiometric
than reference solution Y. (2.2.2, Method If). titration (2.2.20), using 0.1 M sodium hydroxide. Read
pH (2.2.3) the volume added between the 2 pointsof inflexion.
4.5 to 6.0. I mL of 0.1 M sodium hydroxide is equivalent to 41.46 mg of
Dissolve 0.2 g in carbon dioxide-free waterR and dilute to C13H19Br2ClN20"
20 mL with me same solvent. STORAGE
Related substances Protected from lighr.
Liquid chromatography (2.2.29). Prepare the solu'ions IMPURITIES
immediately be/ore use. Otherdete<lable impuriu.s (thefollowing subslances would, if
Test solution Dissolve 50 mg of the substance to be present a' a sufficient levd, bedete<ted by oneor otherof the rests
examined in water R and dilute to 50.0 mL with the same in the monograph. They are limited by thegeneral ac«plana
solvent. criterion for otherlunspedfied impurities and/or by thegeneral
Referena solu.ion (a) Dilute 1.0 mL of the test solution to monograph Substancesfor pharmaceutical use (2034). I. is
100.0 mL with waterR. Dilute 1.0 mL of this solution to therefore not necessary to identify these impun"ties for
10.0 mL with the mobile phase. demonstration of compliance. See also 5.10. Conrrol of impurities
Reference solution (b) In order to prepare impurity B insim, in substanas for pharmaceutical we) A, B C, D, E.
J
dissolve 5 mg of the substance (0 be examined in 0.2 mL of

methanol R, add 0.04 mL of a mixture of 1 volumeof "'~OH
formaldehyde sdution Rand 99 volumes of waterR. Heat at
60°C for 5 min. Evaporate to dryness undera current of
yNH,
nitrogen. Dissolve the residue in 5 mL of water R and dilute B'
to 20.0 mL with the mobile phase.
A. (2-amino-3,5-dibromophenyl)methanol,
Column:
- size: 1= 0.25 m, 0 :;;; 4.0 mm;
- SIa.ianaIY phase: ocuulecylsilyl S17i<a gd for chromatography R
(5 urn).
Mobile phase A mixture of equal volumes of ac:eumirrile R
and a solution prepared as follows: dissolve 1.32 g of
ammonium phosphate R in 900 mL of water R, adjustto
pH 7.0 withphosphori< acid R and dilute to 1000 mL with B. rraO$-4-(6,8-dibromo-l,4-dihydroquin azolin-3(2H)-
water R. yl)cyclohexanol,
Flow rate 1 mlJmin.
Detection Spectrophotometerat 248 nm,
Injection 20 f1L.
Run time 3 times the retention time of ambroxol.
IdentificatWn of impurities Use the chromatogram obtained
with reference solution (b) to identify the peak due to
impurity B.
C. rraO$-4-[[(E)~2-amino-3,5-<1ibromobenzyliden]
Relative retention With reference to ambroxol (retention
= =
time about 9 min): impurity B about 0.6.
aminojcyclchexanol,
System- suitability Reference solution (b): ·00
impurity B and ambroxol.
B'Y'('W 0 ..
Limits:
- unspecified impurities: for each impurity, not more than the yNH,
with reference solution (a) (0.10 per cent), '"
- total: not more than 3 times the area of the principal peak D. ci<-4-[(2-amino-3,5-dibromobenzyl)amino]cyclohexanol,
(0.3 per cent), "'yyCHO
- disregard limir. 0.5 times the area of the principal peak in
yNH,
(0.05 per cent). B'
E. 2-amino-3,5-dibromobenzaldehyde.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'/1£<1
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2022 Amfetamine Sulfate 1-137
Amfetamine Sulfate *** concentrated ammonia R and dilute to 1000 mL with

*** *** acetonitrile R.
Amfetamine Sulphate *** Test solution Dissolve 20.0 mg of the substance to be
(Ph. Eur. monograph 0368) examined in the solvent mixture and dilute to 10.0 mL with
Reference solution (aJ
Dilute 1.0 mL of the test solution to
100.0 mL with the solvent mixture. Dilute 1.0 rnL of this
and enantiomer
Reference sduuon (b) Dissolve 5 mg of I-phenylpropan-2-o1 R
(impurity A) and 5 mg of benzaldehyde R (impurity D) in the
solvent mixture and dilute to 10 mL with the solvent
368.5 60-13-9 mixture. Dilute I rnL of the solution to 100 mL with the
solvent mixture.
Action and use
Releases dopamine; central nervous system stimulant. Column:
- size: 1= 0.15 m, {2) = 4.6 mrn;
P1>E" _ - stationary phase: base-deactivated end-capped ocuulecy/sily/
SIlica gelfor chromatography R (5 pm),
DEFINITION
- temperature; 40 "C.
Bis[(2RSJ-I-phenylpropan-2-amine] sulfate.
Mobile phase:
Content - mobile phase A: solvent mixture;
-' mobile phase B: aatonitn"le R;
CHARACTERS
Appearance Time Mobile phase A MohUe phase B
White or almost white powder. (min) (per cent VIP) (per cent VlJI)
O· I 100 0
Solubility
I . 16 100 ---> 65 0--->35
Freely soluble in water, veryslightly soluble in ethanol
16 - 21 65 ---> 0 35 -> 100
21 - 23 0 100
IDENTIFICATION
First identification: A, B, D. Flow rate 1.5 rnUmin.
Second idenofication: G, D. Detection Spectrophotometer at 257 nm.
A. Optical rotation (see Tests). Injection 20 IlL.
B. Infrared absorption spectrophotometry (2.2.24). Identification of impuriues Use the chromatogram obtained
Comparison amfuomine salfase GRS. with reference solution (b) to identify the peaks due to
C. To 50 rnL of solution S add 5 rnL of scrong sodium impurities A and D.
hydroxide solution Rand 0.5 rnL of benzoyl chloride Rand Relarive resention With reference to amfetamine (retention
shake. Continue to add benzoyl chlodde R in portions of time = about 8 min): impurity D = about 1.6;
0.5 mL, shaking after each addition, until no further =
impurity A about 1.7.
precipitate is formed. Filter, wash the precipitate with System suitability Reference solution (b):
water R, recrystallise twice from a mixture of equal volumes - resolution: minimum 4.0 between the peaks due to
of ethanol (96 per cenv R and waterR, then dry at impurities D and A.
100-105 "C. The crystals melt (2.2.14) at 131 "C to 135 "C.
D. Solution S (see Tests) gives reaction (a) of sulfates - for each impurity, use the concentration of amfetamine
(2.3.1). sulfate in reference solution (a).
TESTS Limits:
Solution S - unspecified impurities: for each impurity, maximum
Dissolve 2.0 g in carbon dioxide-free waterR and dilute to 0.10 per cent;
100 mL with the same solvent. - cota!: maximum 0.5 per cent;
Appearance of solution - reporting threshold: 0.05 per cent.
Solution S is clear (2.2.1) and colourless (2.2.2, Melhod II). Loss on drying (2.2.32)
Optical rotation (2.2.7) Maximum 1.0 per 'cent, determined on 1.000 g by drying in
-0.040 [0 + 0.04 0 (measured in a 2 dm tube), determined on an oven at 105 "C.
solution S. Sulfated ash (2.4.14)
Acidity or alkal1nlty Maximum 0.1 per cent, determined on 1.0 g.
To 25 mL of solution S add 0.1 mL of methyl red solution R. ASSAY
Not more than 0.1 rnL of 0.01 M hydrochloric acid or 0.01 M Dissolve 0.300 g in 30 mL of anhydrous acetic acid R. Titrate
sodium hydroxide is required to change the colour of the with 0.1 ~\1 perch/on'e add, determining the end-point
indicator. potentiomelrically (2.2.20).
Related substances I mL of 0.1 M perchloric acid is equivalent to 36.85 mg
Liquid chromatography (2.2.29). Prepare the solutions of C,.H2S N, O. S.
STORAGE
So/vent mixture Mix 5 mL of lrifluoroaatic acid Rand
900 rnL of waterfor chromatography R, adjust to pH 2.2 with
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1-138 Amidotrizoic Acid Dihydrate 2022
IMPURITIES IDENTIFICATION
Other detectable impurities (thefollowing substances would,· if Firstidentification: A.
present at a sufficient level, be detected by oneor other of the tests Second identification: B, C.
it. the monograph. They arelimited by the general acceptance A. Infrared absorption spectrophotometry (2.2.24).
cruetion for otherlunspedfied impurities and/or by the. general
monograph Substances for pharmaceutical use (2034). It is Comparison amidomzoic add dihydrate CRS.
therefore notnecessary to identify these impuniies for B. Thin-layer chromatography (2.2.27).
demonstraiion of compliance. See also 5. I O. Control of impurities Test solution Dissolve 25 mg of the substance to be
in substances for pharmaceutical use) A, B, C, D. examined in a 3 per cent VIV solution of ammonia R in
methanol R and dilute to 5 mL with the samesolution.
~CH3
V H' OH and enanllomer
Reference soluticn Dissolve 25 mg of amidotrizoic acid
dihydrate CRS in a 3 per cent VIV solution of ammonia R in
methanol R and dilute to 5 mL with the same solution.
A. (2RS)-I-phenylpropan-2-01, Plare TLC silica gel GFZ54 pkue R.
l\1obile phase anhydrous formic acid R, methyl ethyl ketone R,
~CH, rotuene R (20:25:60 VIV/V).
Vo Application 2~.
B. I-phenylpropan-2-one, Drying In air until the solvents have evaporated.
Detection In ultraviolet light at 254 nm.
o Results The principal spot in the chromatogram obtained
~NH, with the test solution is similar in position and size to the
U ,('CH, principal spot in the chromatogram obtained with the
reference solution.
C. Heat 50 mg gently in a small porcelain dish over a naked
C. (2S)-2-amino-l-phenylpropan-l-one (cathinone),
flame. Violet vapour is evolved.
~CHO TESTS
V Appearance of solution
The solution is clear (2.2.1) and colourless (2. Z. 2,
MethodIi).
D. benzaldehyde. Dissolve 1.0 g in dilute sodium hydroxide solution R and dilute
~ PIIE"
to 20 mL with the same solution.
Related substances
Amldotrizoic Acid Dihydrate *** Solvent mixture Dissolve 0.250 g of sodium hydroxide Rand
*** *** 0.860 g of sodium dihydrogen phosphate R in 50 mL of water R
*** and dilute to 1000 mL with the same solvent.
examined in 10.0 mL of me solvent mixture with the aid of
o
)l. ,*:::H'
I
0
)l.' 2 H,O
ultrasound.
H,C ~ ~ CH, 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
I solution to 10.0 mL with the solvent mixture.
Reference solution (b) Dilute 1.0 mL of reference solution (a)
650 5097t-1l-5 to 10.0 mL with the solvent mixture.
Reference solution (c) Dissolve the contents of a vial of
Action and use amidotrizoic acidfor system SUilability CRS (impurities A, BJ C
Iodinated contrast medium.
and D) in 1.0 mL of the solvent mixture.
Preparation Column:
lVlegluJ:llne Amidotrizoate Injection - size: 1 = 0.25 m, 0 = 4.6 nun;
PIlE" _ _ ~ __ ~ ~ _ - stationary phase: end-capped lXtaduy/si1y1 silica gelfor
chromatagrapi!)l R (5 urn).
DEFINITION
Mobile phase Dissolve 3.4 g of retrabutylammonium hydrogen
3,5-Bis(acetylamino)-2,4,6-triiodobenzoic acid dihydrate. sulfate R in a mixture of 230 mL of acetonitrile Rand 770 mL
Content of water R.
98.5 per cent '0 101.0 per cent (dried substance). F!qw rate 1.0 mllmin.
CHARACTERS Deteaion Spectrophotometer at 236 run.
Appearance Injection 20~.
White or almost white, crystalline powder. Run time 4 times the retention time of amidotrizcic acid.
Solubility Identification of impun'lies Use the chromatogram supplied
Very slightly soluble in water and in ethanol (96 per cem). with omidotnzoic acidfor system suitability CRS and the
It dissolves in dilute solutions of alkali hydroxides.
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2022 Amidotrizoic Acid Dihydrate 1-139
chromatogram obtained with referencesolution (c) to identify through a sintered-glass filter (2.1.2) and wash the filter with
the peaks due to impurities A, B, C and D. several quantities of warer R. Collect the filtrate and
Relativeretention Withreference to amidotrizoic acid washings. Add 40 mL of dihue su/furic add R and titrate
(retention time = about 5 min): impurity B = about 0.8; immediately with 0.1 1\1 silvernitrate. Determine the
impurity C = about 0.9; impurityA = about 1.4; end-point potentiomenically (2.2.211).
=
impurity D about \.8. 1 mL of 0.1 M silvernitrate is equivalent to 20.47 mg of
System suitability: ClIH913N204'
- resolution: minimum 1.5 between the peaks due £0 STORAGE
impurities Band C in the chromatogram obtained with Protected from light.
- signal-to-noise ratio: minimum 25 for the principal peak in IMPURITIES
me chromatogram obtained with reference solution (b). Specified impun·ties A, B, D.
Limits: Otherdetectable impunties (thefollowing substances would, if
- impun'ty B: not more than the area of the principal peak in present at a sufficient level, be detected ~ oneor other of the tests
the chromatogram obtained with referencesolution (a) in the monograph. They are limited by thegeneral acceptance
(0.1 per cent); cmenon for otherlunspmfied impurities and/or by the general
- impurities A, D: for each impurity, not more than the area monograph Substances for pharmaceutical use (2014). II is
of the principal peak in the chromatogram obtained with therefore not necessary to identify these impwities for
reference solution (b) (0.01 per cent); demonstration of romp/janu. See also5.10. Control of impurities
- unspecified impun"ties: for each impuri£y, not more than in substances for pharmaceutical use) C, E.
0.5 times the area of me principal peak in the
,*~H,
(0.05 per cent);
I 0
~~cH,
- total: not more than 1.5 rimes the area of the principal
peakin the chromatogram obtainedwith reference H,N ""
solution (a) (0.15 per cent); I
the chromatogram obtainedwith reference solution (a) A. 3-(acetylamino)-5-amino-2,4,6-triiodobenzoic acid,
(0.03 per cent), except for the peaks due to impurities A
and D.
Halides expressed as chlorides (2.4.1)
Masimum 150 ppm.
Dissolve 0.55 g in a mixture of 4 mL of dilute sodium
hydroxide solution Rand 15 mL of water R. Add 6 mL of
dilule nitric add R and filter.
B. 3,5-bis(acetylamino)-2,4-diiodobenzoic acid,
Free aromatic amines
Maintain thesolutions and reagents in iced water, protected from
bright light To 0.50 g in a 50 mL volumetric flask add
15 mL of water R. Shake and add 1 mL of dl1ule sodium
hydroxide solution R. Cool in iced water, add 5 mL of a
freshly prepared 5 gIL solution of sodium nitrile R and 12 mL
of dilute hydro<:hloric acid R. Shake gently and allow to stand
for exactly 2 min after adding the hydrochloric acid. C. 3,5-bis(acetylamino)-2,6-diiodobenzoic acid,
Add 10 mL of a 20 gIL solution of ammonium sulfamale R.
Allow to standfor 5 min, shaking frequently, and add
0.15 mL of a 100 gIL solution of a-naphthol R in ethanol
(96 per unt) R. Shake and allow to stand for 5 min. o
,*CO,H,
I"" 0
Add 3.5 mL of buffer solution pH 10.9 R, mix and dilute to
50.0 mL with water R. The absorbance (2.2.25), measured
H3C~~ , ~~' ""
within 20 min at 485 nm using as the compensation liquid a

solutionprepared at the same time and in the same manner
D. 3-( acetylamino)-5-[(iodoacetyl)antino]-2,4,6-
but omitting the substance to be examined, is not greater
triiodobenzoic acid,
than 0.30.
4.5 per cent to 7.0 per cent, determined on 0.500 g by
drying in an oven at 105 DC.
Sulfatedash (2.4.11)
ASSAY
To 0.150 g in a 250 mL round-bottomed flask add 5 mL of E. 3-(acetylantino)-5-(diacetylamino)-2,4,6-triiodobenzoic
srong sodium hydroxide solnt.m R, 20 mL of waterR, 1 g of acid.
zinc powder R and a few glass beads. Boil under a reflux _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE",
condenser for 30 min. Allow to cool and rinsethe condenser
with 20 mL of water R, adding the rinsings to the flask. Filter
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1-140 Amikacin 2022
Amikacin .*.
••• •••
••*
TESTS
pH (2.2.3)
(Ph. Eur. monograph 1289) 9.5 to 11.5.
Dissolve 0.1 g in carbon dioxide-free waterR and dilute to
HO~
Nil, 0 + 97 to + 105 (anhydrous substance).
H,N~o 0 ~N~NH' Dissolve 0.50 g in waterR and dilute to 25.0 mL with the
same solvent.
o OHq HOH
OH HO-·
Related substances
HO ~ Liquid chromatography (2.2.29).
OH 0 NH2
Test solution Dissolve 25 mg of me substance to be
examined in mobile phaseA and dilute to 50.0 mL with
585.6 37517-28-5 mobile phase A.
Reference solu'ion (a) Dilute 1.0 mL of the test solution to
Action and use
100.0 mL with mobile phase A.
Aminoglycoside antibacterial.
Reference solu'ion (b) Dilute 1.0 mL ofreference solution (a)
PhE" _
to 10.0 mL with mobile phase A.
DEFINITION Reference solution (c) Dissolve 5 mg of amikacin for system
6-0-(3-Amino-3-deoxy-<t.-o-g1ucopyranosyl)-4-0-(6-amino-6- suitability CRS (containing impurities A, B, F and H) in
deoxy-c-n-glucopjranosyl) -I-N-[(2S)-4-amino-2- mobile phase A and dilute to 10 mL with mobile phase A.
hydroxybutanoyl]-2-deoxy-D-streptamine. Reference solution (d) Dissolve 5.0 mg of amikacin
Antimicrobial substance obtained from kanamycin A. impurity 1 CRS in mobile phase A and dilute to 20.0 mL with
mobile phase A. Dilute 1.0 mL of the solution to 100.0 mL
Semi-synthetic productderived from a fermentation product.
with mobile phase A.
Content
Column:
- size: 1= 0.25 m, 0 = 4.6 mm;
CHARACTERS - stationary phase: end-eapped aaadecylsilYl silica gelfor
Appearance chromatography R (5 um),
Whiteor almost while powder. - temperature: 40°C.
Soluhility Mobile phase:
Sparingly soluble in water, slightly soluble in methanol, - mobile phase A: a mixtuce prepared with carbon dioxide-free
practically insoluble in acetone and in ethanol (96 per cent). water RJ containing 1.8 gIL of sodium oaanesulftmate R,
20 gIL of anhydrous sodium sulfate Rl, 1.4 per cent VIV of
IDENTIFICATION
tetrahydrofuran R, and 5 per cent VIV of 0.2 M potassium
A. Infrared absorption spectrophotometry (2.2.24}.
dihydrogen phosphate R previously adjusted to pH 3.0 with
Comparison amikadn CRS. diluu phospho", acidR; degas;
B. Thin-layer chromatography (2.2.27). - mobile phaseB: a mixture prepared with carbon dioxide-free
Test solution Dissolve 25 mg of the substance to be water R, containing 1.8 gIL of sodium oaanesulfonace R,
examined in water R and dilute to 10 mLwith the same 28 gIL of anhydrous sodium sulfate Rl, 1.4 per cent VIV of
solvent. tetrahydrofuran R, and 5 per cent VIV of 0.2 M potassium
Reference solution (a) Dissolve 25 mg of amikacin CRS in dihydrogen phosphate R previously adjusted to pH 3.0 with
water R and dilute to 10 mL with the same solvent. dilute phosphoric acidR; degas;
Reference solution (b) Dissolve 5 mg of kanamycin Time Mobile phase A Mobile phase B
monosuljate CRS in 1 mL of the test solution and diluteto (mln) (per cent VIJI) (per cent VIV)
10 mL with water R. 0-3 100 0
Plate TLC silica gelplate R. 3 - 38.0 100 -+ 30 0-+70
J.'AobiJe phase methylene chloride R, ammonia R, methanol R 38.0 - 38.1 30 ..... 0 70 -+ 100
(25:30:40 VIV1V). 38.1-68 0 100
Application 5 ~L.
Development Over 3/4 of the plate. Flow rate 1.0 mlJmin.
Drying In air. Post-cdumn solution Mixture of 1 volwne of carbonate-free
Detection Spraywith ninhydrin SOluri011 Rl and heat at sodium hydroxide sobmon R and 24 volumesof previously
110 'C for 5 min.
degassed carbon dioxide-free water R, which is added in a
pulseless manner to the column effluent using a 375 ilL
System su;tabih·ty Reference solution (b): polymeric mixing coil.
- the chromatogram shows 2 clearly separated spots.
Flow rate ofpost-column solutWn 0.3 mUmin.
with the test solution is similar in position, colour and size to Detection Pulsed amperometric detector or equivalent with a
the principal spot in the chromatogram obtained with gold indicator electrode, a silver-silver chloride reference
reference solution (a). electrode, and a stainless steel auxiliary electrode which is the
cell body, held at respectively + 0.05 V detection, + 0.75 V
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2022 Amikacin 1-141
oxidation and - 0.15 V reduction potentials, with pulse in the mobile phase; peak splitting may be observed when
durations according to the instrument used. the retention time becomes too short;
Injection 20 ~L. - repeatability: maximum relative standard deviation of
1.5 per cent after 6 injections.
with amihadn for systemsuitabl1ity CRS and the Calculate the percentage content of C22H43NjOl3 taking into
chromatogram obtained with reference solution (e) to identify account the assigned content of amikacin CRS.
the peaks due to impurities A, B, F and H; use the IMPURITIES
chromatogram obtained with reference solution (d) to Specified impun·ues A, BJ F, H, 1.
identify the peak due to impurity I.
Otherdetectable impurities (the following substances 'WOuldJ if
Relative retention With reference to amikacin (retention present at a sufficient Ieve/J be deteaed by one or otherof the tests
=
time about 28 min): impurity I = about 0.13; in the monograph. They are limited by the general acceptance
= =
impurity F about 0.92; impurity B about 0.95; criterion for other/unspecified impulilies and/or by the general
= =
impurity A about 1.62; impurity H about 1.95. monograph Substances for pharmaceutical use (2034). 1/ is
System suitability Reference solution (c): therefore not necessary w identifythese impurities for
=
- peak-eo-vaHey ratio: minimum 5, where H p height above demonstration of compliance. See also5.10. Control of impuruies
the baseline of the peak due to impurity Band in substances for pharmaceutical use) CJ D, EJ G.
H v = height above the baseline of the lowest point of the
curve separating this peak from the peak due to amikacin; HO
if necessary, adjust the volume of tetrahydrofuran in the
mobile phase.
Cakulation ofpercentage contents:
- for impurity I, use the concentration of impurity I in
reference solution (d); OH o OH~
HO"
- for impurities other than I, use the concentration of HO ', 0
amikacin in reference solution (a).
OH 0 HNh
Limits:
- impurities A, BJ P, H, I: for each impurity, maximum OH N~
0.5 per cent;
- any other impun·ty: for each impurity, maximum A. 4-0-(3-amino-3-deoXY-<X-D-g1ucopyranosyl)-6-D-(6-amino-
0.5 per cent; . ti-decxy-c-n-glccopyranosylj-l-N; [(2S)-4-amino-2-
- total: maximum 1.5 per cent; hydroxybutanoyl]-2-deoXY-L-streptamine,
- reponing threshold: 0.1 per cent.
HO
Water (2.5.1Z)
Maximum 0.5 per cent, determined on 1.0 g. HO 0 HN
ASSAY
o OH~· H"elH
OH HO··
liquid chromatography (2.2.29). HO \ 0
Test sokuion Dissolve 50.0 mg of the substance to be OH 0 HN---{
the mobile phase.
H~
OH NH2
Reference solution Dissolve 50.0 mg of amikacin CRS in the

mobile phase and dilute to 10.0 mL with the mobile phase. B. 4-0-(3-amino-3-deoXY-<X-D-g1ucopyranosyl)-6-D-(6-amino-
6-deoxy-<X-D-g1ucopyranosyl)-1,3-N-bis[(2SJ-4-amino-2-
Column: hydroxybutanoylj-2-deoxy-L-streptamine,
=
- size: I 0.25 ffi, 0 = 4.6 mm;
- s<aMnary phase: end-capped oaadecylsilyl silica gelfor
ehromalOgraphy R (5 pm):
·H'N~O
Mobilephase A mixture prepared with carbon dioxide-free HO H
HN
~H
'~o 0
water RJ containing 1.8 gIL of sodium ocumesalfonaie R,
20 gIL of anhydrous sodium sulfate R1, 5.8 per cent VIV of
acetonitrile R 1, and 5 per cent VIV of O. 2 M potassium
HO
OH o HO·· OHH rNH,
.
dihydrogen phosphate R previously adjusted to pH 3.0 with
dilutephosphoric acid R; degas. OH 0 N~
FlOO! rate 1.0 mUmin.
C. 4-0-(6-amino-6-deoXY-<X-D-g1ucopyranosyl)-6-D-[3-[[(2SJ-
Detection Spectrophotometer at 200 nm. 4-amino-2-hydroxybutanoyl]amino]-3-deoXY-<X-D-
Injeelion 20 ~L. glucopyranosyl]-2-deoXY-D-streptamine,
Run time 1.3 times the retention time of amikacin.
Retention time Amikacin = about 30 min.
System suitability Reference solution:
amikacin; if necessary, adjust the amount of acetonitrile Rl
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1-142 Amikacin Sulfate 2022
Ho,C~NH,
HO~ H OH
NH,
H,N~o 0 ,Nil,
I. (2S)-4-amino-2-hydroxybutaooic acid.
H::.~.K
_ ~ IM"
OH 0
HO
OH 0
~NH2
Amikacin Sulfate ***
D.6-G-(3-amino-3-deoxy-«-o-glucopyraoosyl)-4-G-(6-amino- *** ***
6-deoXY-«-D-glucopyraoosyl)-2-deoxy-o-streptamine Amikacin Sulphate ***
(kanamycin), (ph. Bur. monograph 1290)
~~~~"'
HO
o 0
o OH-
0 OH~
HO-· OH
o OH~
HO--
HOH
HO \ HO ..
OH 0 N~ OH 0 NH2
E. 4-G-(3-amino-3-deoxy-«-D-glucopyranosyl)-6-G-[6-[[(2S)- 782 39831-55-5

4-amino-2-hydroxybutanoyl]amino]-6-deoxy-«-o-
glucopyranosyl]-2-deoXY-L-streptamine, Action and use
Aminoglycoside antibacterial.
~~~~ ""~"'
Preparation
Amikacin Injection
PIlE" _ _ ~~ ~ _
o 0 OH~HOH DEFINITION
OH HO·· 6-G-(3-Amino-3-deoxy-«-D-glucopyraoosyl)-4-G-(6-amino-6-
HO .. deoxy-«-D-glucopyranosyl)-I-N-[(2S)-4-amino-2-
OH 0 NH2 hydroxybutaooyl]-2-deoXY-D-Stleptamine sulfate.
Antimicrobial substance obtained from kanamycin A.
F. 6-G-(3-amino-3-deoXY-«-D-glucopyranosyl)-4-G-[6-[(2S)- Semi-synthetic product derived from a fermentation product.
4-amino-2-hydroxybutanoyl]amino-6-deoxy-«-D-
glucopyraoosyl]-I-N-[(2S)-4-amino-2-hydroxybutanoyl]-2- Content
deoxy-n-streptamine, 96.5 per cent to 102.0 per cent (dried substaoce).
CHARACTERS
H'N~HO~O' 1X~--
Appearance
H~
.NH, Solubility
HN' Freely soluble in water, practically insoluble in acetone and
o OH~ H' OH in ethanol (96 per cent).
OH HO··
IDENTIFICATION
HO ..
A. Infrared absorption spectrophotometry (2.2.Z4).
OH a NHl
Compo,;,on amikacin sulfat< CRS.
G.6-G-(3-amino-3-deoxy-«-D-glUcopyranosyl)-4-G-(6-amino- B. Thin-layer chromatography (2.2.27).
e-deoxy-c-n-glucopyranosyl)-I-N-[(2R)-4-amino-2- Test solution Dissolve 25 mg of the substance to be
hydroxybutaooyl]-2-deoXY-D-Stleptamine, examined in water R and diluteto 10 rnL with the same
solvent. "
H'N~HO~O' 1X~NH,
Re/erenu solution (a) Dissolve 25 mg of amikadn
sulfat< CRS in warer R and dilute to 10 mL with the same
H~
solvent.
HN' '-./ Reference solutinn (b) Dissolve 5 mg of kanamycin
o OH~' H '<JH monosulfat< CRS in I mL of the test solution and dilute to
OH HO··
10 mL with water R.
HO •
Plat< TLC silica gelplat< R.
NH2 0 NH:!
Mobile phase methylene chloride R) ammonia R, methanol R
(25:30:40 VIV/V).
H.6-G-(3-amino-3-deoxy-«-D-glucopyraoosyl)-1-N-[(2S)-4-
amino-2-hydroxybutaooylj-4-G-(2,6-diamino-2,6-dideoxy- Application 5~.
(x-D-glucopyranosyl)~2-deoxy-o-streptamine, Development Over 3/4 of the plate.
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2022 Amikacin Sulfate 1-143
Drying In air. Post-column solution Mixture of 1 volume of carbonate-free

Detection Spray with ninhydrin solution RJ and heat at sodium hydroxide solution Rand 24 volumes of previously
110 °C for 5 min. degassed carbon dioxide-free water R, which is added in a
System suitability Reference solution (b): pulseless manner to the column effluent using a 375 ~L
- the chromatogram shows 2 clearly separated spots. polymeric mixing coil.
Results The principal spot in the chromatogram obtained Flow rateof post-column solution 0.3 mUmin.
with the test solution is similar in position, colour and size to Detection Pulsed amperometric detector or equivalent with a
the principal spot in the chromatogram obtained with gold indicator electrode, a silver-silver chloride reference
reference solution (a). electrode, and a stainless steel auxiliary electrode which is the
C. It gives reaction (a) of sulfates (2.3.1). cell body, held at respectively + 0.05 V detection, + 0.75 V
oxidation and - 0.15 V reduction potentials, with pulse
TESTS durations according to the instnunent used.
pH (2.2.3) Injection 20 ~L
2.0 to 4.0.
Dissolve 0.1 g in carbon dioxide-free waterR and dilute to with amikacinfor system suitability CRS and the
10 mL with the same solvent. chromatogram obtained with reference solution (c) to identify
Specific optical rotation (2.2.7) the peaks due to impurities A, B, F and H, use the
+ 76 to + 84 (dried substance). chromatogram obtained with reference solution (d) to
Dissolve 0.50 g in water R and dilute to 25.0 mL with the identify the peak due to impurity I.
same solvent. Relative retention With reference to amikacin (retention
Related substances =
time about 28 min): impurity I = about 0.13,
Liquid chromatography (2.2.29). = =
impurity F about 0.92; impurity B about 0.95;
Test solution Dissolve 33 mg of the substance (0 be = =
impurity A about 1.62; impurity H about 1.95.
examined in mobile phase A and dilute to 50.0 mL with System suitability Reference solution (c):
mobile phase A. =
- peak-to-'lJal/ey ratio: minimum 5, where Hp height above
the baseline of the peak due to impurity B and
curve separating this peak from the peak due to amikacin;
Reference solution (b) Dilute 1.0 mL of reference solution (a) if necessary, adjust the volume of tetrahydrofuran in the
to 10.0 mL with mobile. phase A. mobile phase.
Reference solution (c) Dissolve 5 mg of amikacinfor system Calculation of pemmrage centents:
suitabl7.;y CRS (containing impurities A, B, F and H) in - for impurity I, use the concentration of impurity I in
mobile phase A and dilute to 10 mL with mobile phase A. reference solution (d),
Reference solution (d) Dissolve 6.6 mg of amikacin - for impurities other than I, use the concentration of
impurity I CRS in mobile phase A and dilute to 20.0 mL with amikacin sulfate in reference solution (a).
mobile phase A. Dilute 1.0 mL of the solution to 100.0 mL Limits:
with mobile phase A. - impurities A J B, F, H, I: for each impurity, maximum
Column: 0.5 per cent,
=
- size: / = 0.25 m, 0 4.6 rnm; - atry otherimpun"ty: for each impurity, maximum
- stationary phase: end-capped actadecylsily1 silica gelfor 0.5 per cent,
chroma/(Jgraphy R (5 ~m); - total: maximum 1.5 per cent;
- temperature: 40°C. - reporting threshold: 0.1 per cent.
Mobile phase: Sulfate
- mobile phase A: a mixture prepared with'carbon dioxide-free 23.3 per cent to 25.8 per cent (dried substance).
waterR, containing 1.8 gIL of sodium octanesulfonate R,
Dissolve 0.250 g in 100 mL of water R and adjust the
solution to pH 11 using concentrated ammonia R.
tetrahydrofuran R, and 5 per cent VIV of 0.2 M potassium
Add 10.0 mL of 0.1 M barium chloride and about 0.5 mg of
dihydrogm phosphate R previously adjusted to pH 3.0 with
phthalein purple R. Titrate with 0.1 M sodium edetate adding
dilute phosphoric add R, degas;
50 mL of ethanol (96 per cent) R when the colour of the
- mobil» phase B: a mixture prepared with carbon dioxide-free
solution begins to change and continue the titration until the
waterR, containing 1.8 gIL of sodium ocumesulfonau R, violet-blue colour disappears.
tttrahydrofuran R, and 5 per cent VIV of 0.2 lWpotassium I mL of 0.1 M barium chloride is equivalent to 9.606 mg of
dihydrogen phosphate R previously adjusted to pH 3.0 with sulfate (S04).
dilute phosphoric acidR; degas; Loss on drying (2.2.32)
Tim. Mobile phase A Mobile phase B an oven at 105°C at a pressure not exceeding 0"7 kPa
(min) (per cent VIl1 (per cent V/JI) for 3 h.
0-3 100 o Pyrogens (2.6.8)
3 - 38.0 100 ---> 30 0--->70 If intended for use in the manufacture of parenteral
38.0·38.1 30 ---> 0 70 ---> 100 preparations without a further appropriate procedure for the
38.1-68 o 100 removal of pyrogens, it complies with the test for pyrogens.
Inject per kilogram of the rabbit's mass 5 mL of a solution
Flow rate 1.0 mUmin. containing 25 mg of the substance to be examined in water
for injections R.
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1-144 Amikacin Sulfate 2022
H,N~~~OHR".~~
ASSAY
the mobile phase.
OH
HOH
o
HO--
Reference solution Dissolve 37.4 mg of amikacin CRS in the
mobile phase and dilute to 10.0 mL with the mobile phase. HO -, 0
Column:
OH 0 HN~
=
- size: I ::;;; 0.25 m, 0 4.6 nun; H~
OH NH2
- stationary phase: end-eapped IXtadecylsilyl silica gelfor
chromatography R (5 pm}; . B. 4-0-(3-amino-3-deoxy-«-o-glucopyranosyl)-6-0-(6-amino-
- temperature: 40°C. e-deoxy-c-n-glucopyranosyl)-1,3-N-bis [(2S)- 4-amino-2-
Mobile phase Mixture containing 1.8 gIL of sodium hydroxybutanoylj-2-deoXY-L-streptamine,
IXtanesulfonale R, 20 gIL of anhydrous sodium sulfare Rl, o H0
5.8 per cent VIV of acetonitrile Rl, and 5 per cent VIVof 1
0.2 M potassium dihydrogen phosphare R previously adjusted to H,N~O
pH 3.0 with dilute phosphoric acid R; degas. HO'-H
HN
~H
Flow rate 1.0 mUmin. '~o 0 ..NH,
OH
o OHp
HO--
Injedon 20 ~L.
Run time 1.3 times the retention time of amikacin.
HO OH
.
0 'NHa
=
Retention time Amikacin about 30 min.
C. 4-0-(6-amino-6-deoxy-«-o-glucopyranosyl)-6-D-[3-[[(2S)-
4-amino-2-hydroxybutanoyl]aminoj-3-deoxy-«-o-
glucopyranosylj-2-deoxy-D-streptamine,
amikacin; if necessary, adjust the amount of cuetonitrile Rl
in the mobile phase; peak splitting may be observed when
the retention time becomes too short;
- repeatability: maximum relative standard deviation of . HO~
1.5 per cent after 6 injections. NH,
Calculate the percentage content ofC2zl147Nj021S2 taking H,N~O 0 ,NH,

into account the assigned content of amikacitl GRS and a OH 0 H:,.K
correction factor of 1.335"
STORAGE HO
OH 0
K N~
In an airtight container. If the substance is sterile, the
container is also sterile and tamper-evident. D.6-D-(3-amino-3-deoxy-«-o-glucopyrnnosyl)-4-0-(6-amino-
IMPURITIES 6-deoXY-«-D-glucopyranosyl)-2-deoXY-D-streptamine
Specified impurilies A, B, F, H, 1. (kanamycin),
Oriler detectable impurities (rile following subS/ances UJOuld, if HO
present at a sufficient. level, be detected by one or otherof the tests
in the monograph. They arelimited by the general acceptance
,n"ten"on for othethmspecified impurities and/or by the general
monograph Substances for pharmaceutical use (2034). II is
therefore not neassary UJ identify these impun"lie.s for o
OH
0 OHp
HO·-
demonstration of compliance. See also 5.10. Contrd of impun'ties
in substances for phannaceuticaJ use) G, D, E, G.
.
HO OH
. 0 'NHa
HO E. 4-D-(3-amino-3-deoxy-«-o-glucopyranosyl)-6-D-[6-[[(2S)-
o 4-amino-2-hydroxybutanoyljaminoj-6-deoxy-«-o-
HN N~ glucopyranosyl]-2-deoxy-L-streptamine,
HO
OH o HO--OHR
HO -, 0
OH 0 HN----{
H~NH:! o
OH OOHpHOH HO--
A. 4-D-(3-amino-3-deoXY-«-D-glucopyranosyl)-6-D-(6-amino-
HO .'N~
OH 0
6-deoXY-«-D-glucopyranosyl)-I-N-[(2S) -4-amino-2-
hydroxybutanoylj-2-deoxy-L-streptamine, F. 6-0-(3-amino-3-deoxy-«-o-glucopyranosyl)-4-D-[6-[(2S)-
4-amino-2-hydroxybutanoyl]amino-6-deoxy-«-D-
glucopyranosyl]-I-N-[(2S)-4-amino-2-hYdroxybutanoyl]-2-
deoxy-n-srreptamlne,
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2022 Amiloride Hydrochloride Dihydrate 1-145
HO Solubillty
o 0
Slightlysoluble in water and in anhydrous ethanol, practically
NH, insoluble in heptane.
IDENTIFICATION
OH
o
HO--
OHp HOH First identification: A, c, D.
Second identification: B, C, D.
HO ..
OH 0 NH2 A. Infrared absorption spectrophotometry (2.2.24).
Comparison ami/oride hydrochloride CRS.
G.6-Q-(3-amino-3-deoxy-«-D-glucopyranosyl)-4-0-(6-amino- B. Thin-layer chromatography (2.2.27).
ri-deoxy-c-n-glucopyranosylj-J-N. [(2R)-4-amino-2- Test solution Dissolve 5 mg of the substance to be examined
hydroxybutanoyl]-2-deoxy-o-streptamine, in methanol Rand dilute to 10 mL with the same solvent.
HO Reference solution Dissolve 5 mg of omdonde
hydrochloride CRS in methanol R and dilute to 10 mL with the
same solvent.
Plate TLC silica gel Fm plate R.
HO 0 HN Mobile phase concentrated ammonia R, propanol R
OH
o HO-~
OHp' H'OH (30:10 VIII).
Appliauion 5 JIl..; the volume may be adapted according to
HO ..
the type of plate used.
NH2 0 NH2
H.6-Q-(3-amino-3-deoxy-«-o-glucopyranosyl)-I-N-[(2S)-4- Drying In ait.
amino-2-hydroxybutanoyl]-4-Q-(2.6-diamino-2,6-dideoxy- Detection Examine in ultraviolet light at 254 nm.
iX-D-glucopyranosyl)-2-deoxy-D-streptamine, Results The principal spot in the chromatogram obtained
Ho,C~NH, principal spot in me chromatogram obtained with the
H OH reference solution.
C. Dissolve 25 mg of the substance to be examined in
1. (2S)-4-amino-2-hydroxybutanoic acid. water R and dilute to 10 mL with the same solvent. 2 mL of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEor the solutiongives reaction (a) of chlorides (2.3.1); acidify
with 0.5 mL of di/ute acetic acidR, instead of diluu nitric
acid R.
D. Water (see Tests).
Amiloride Hydrochloride Dihydrate TESTS
Free acid
(ph. Bur. monograph 0651) Dissolve 1.0 g in a mixrure of 50 mL of methanol Rand
50 mL of wacn: R and titrate with 0.1 M sodium hydroxide,
determining the end-point potentiometrically (2.2.21}).
Not more than 0.3 mL of 0.1 M sodium hydroxide is required
to reach the end-point.
Related substances
CoH,C1,N,O,2H,O 302.1 1744()'83-4 Solutwn A Dissolve 2.16 g of 'odium dihydrogen phosphate
monohydrate R in 850 mL of waterfor chromatography R,
Action and use adjust to pH 3.0 with phosphoric acid R and dilute to
Sodium channel blocker; potassiwn-sparing diuretic. 1000 mL with waterfor chromatography R.
Preparations Test solution Dissolve 20 mg of the substance to be
Amiloride Tablets examined in I mL of methanol R and dilute to 10.0 mL with
Co-amilofruse Tablets solution A.
Co-amilozide Oral Solution Reference solution (a) Dissolve 2 mg of amiloride
Co-amilozide Tablets impurity A CRS in 0.5 mL of methanol R, add 0.5 mL of the
test solution and dilute to 10 mL with solution A.
PhEor _
Reference solutwn (b) Dissolve 4 mg of ami/oride for peak
DEFINITION identification CRS (containing impurity C) in 0.5 mL of
3,5-Diamino-6-cWolO-N-(diaminomethylidene)pyrazine-2- methanol R and dilute to 2 mL with solution A.
carboxamide hydrochloride dihydrate. Reference solution (c) Dilute 1.0 mL of the test solution to
Content 100.0 mL with solution A. Dilute 1.0 mL of this solution to
98.0 per cent to 101.0 per cent (anhydrous substance). 10.0 mL with solution A.
CHARACTERS Column:
- size: 1= 0.125 m, 0 = 4.0 mm;
Appearance
- stationary phase: base-deactivated octylsilyl silica gelfor
Pale yellow or greenish-yellow powder.
chromatography R (5 pm);
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1-146 Aminobenzoic Acid 2022
Mobile phase Dissolve 0.8 g of sodium hexanesulfonate
monohydrate R in a mixture of 80 m.L of acetonitrile Rl and
920 mL of solution A.
B. 3,S-diamino-6-chloropyrazine-2-carboxylic acid,
Daeaion Spectrophotometer at 210 nID.
Injection 20~.
Run time Twice the retention time of amiloride.
Identification of impurities Use me chromatogram obtained
with reference solution (a) to identify the peak due to C. 3-amino-6-chloro-N-(diaminomethylidene)-5-
impurity A; use the chromatogram supplied with amiloride for hydroxypyrazine-2-carboxamide.
j>Mk identification eRS and the chromatogram obtained wilh _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"'
reference solution (b) to identify the peak due to impurity C.
Relative retention With reference to amiloride (retention
time = about 10 min): impurity C = about 0.5;
Sysrem suitability Reference solution (a):
Aniinobenzoic Acid
- resolution: minimum 3.0 between the peaksdue to (4-Aminobenzoic Acid, Ph. Eur. monograph 1687)
impurity A and amiloride.
Calculation of percentage contents: [""'yco,H
- for each impurity) use the concentration of amiloride
hydrochloride dihydrate in reference solution (c). H,NJV
Limits:
- impuniy C: maximum 0.2 per cent; C,H,NO, 137.1 150-13-0
0.10 per cent; Action and use
- total: maximum 0.4 per cent; Skin protective.
- repuning .hreslwld: 0.05 per cent. PhE<; ~
Water (2.5.12)
11.0 per cent to 13.0 per cent, determined on 0.200 g.
DEFINITION
4-Aminobenzoic acid.
Maximum 0.1 per cent, determined on 1.0 g. Content
ASSAY
CHARACTERS
Dissolve 0.200 g in a mixture of 5.0 mL of O. 01 M
hydrodJlori< arid and 50 rnL of ",hanoJ (96 per cem) R. Carry Appearance
out a potentiometric titration (2.2.2lJ), using 0.1 M sodium White or slightly yellow, crystalline powder.
hydroxide. Read the volume added between the 2 points of Solubility
inflexion. Slightly soluble in water, freely soluble in ethanol
1 rnL of 0.1 M sodium hydroxide is equivalent to 26.61 mg (96 per cent). It dissolves in dilute solutions of alkali
of C6H,CI,N,O. hydroxides.
STORAGE IDENfIFICATlON
Protected from light. Fint identification: B.
Second identifica.ion: A, G.
IMPURITIES
Specified impurilies G. A. Melting point (2.2.14): 186°C to 189 °C.
Otherdetetlable impuniies (the following subslances would, .f B. Infrared absorption spectrophotometry (2.2.24).
present at a sufficient level, be detected by oneor other of me tes" Comparison 4-aminobenzoic add CRS.
in me monograph. They are limited by the general aeceplance C. Thin-layer chromatography (2.2.27).
criterion for othethmspecified impurities and/or by me general Test solution Dissolve 20 mg of the substance to be
monograph Subslances for pharmaceu.ical use (2034). It is examined in methanol R and dilute to 20 mL with the same
therefore not necessary to identify these imPurities for solvent.
demonstration of compliance. See also 5.10. Control of impurities
Reference solution (a) Dissolve 20 mg of 4-aminobenzoic
in substanas for pharmaceutical use) A, B.
acid GRS in methanol R and dilute to 20 rnL with the same
solvent.
Reference solution (b) Dissolve 10 mg of 4-nitrobenzoic arid R
in 10 mL of reference solution (a).
Plate Suitable silica gel with a fluorescent indicator having
an optimal intensity at 254 nm as the coatingsubstance.
A. methyl 3J5-d.iamino-6-chtoropyrazine-2-carboxylate, Mobile phase glacial acetU acid RJ hexane R, medrylem
chloride R (5:20:75 VIVIV).
Application 1 ~L.
Deudopmeni Over a path of 10 em.
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2022 Aminobenzoic Acid 1-147
Drying In air. Test solution Dissolve 1.000 g of the substance to be

Detection Examine in ultraviolet light at 254 run. examined in 10.0 mL of an 84 gILsolution of sodium
SystemsuitabJ1ity The chromatogram obtainedwith hydroxide R and extract with 2 quantities, each of lOrn!.., of
reference solution (b) shows 2 clearlyseparatedspots. methylene chloride R. Combine and wash with 5 mL of
water Rj filter through anhydrous sodium sulfate R. Wash the
filter with methylene chloride R. Evaporate in a water-bath at
with the test solution is similar in position and size £0 the
50-60"C to obtain a volume of about 1-5 mL. Add 1.0 mL
of the internal standard solution and diluteto 10.0 mL with
solution (a).
methylene chloride R.
TESTS Reference solution (a) Dissolve 20.0 mg of aniline R in
Appearance of solution methylene chloride R and dilute to 100.0 mL with the same
The solution is clear (2.2.1) and not more intensely coloured solvent.
than referencesolution n, (2.2.2, Mezhod II). Reference solution (b) Dissolve 20.0 mg of p-toluidine R in
Dissolve 1.0 g in ethanol (96 per ccn!! R and dilute to 20 mL methylene chloride R and dilute to 100.0 mL with the same
with the same solvent. solvent.
Related substances Reference solution (c) Dilute 0.50 mL of reference
Liquid chromatography (2.2.29). solution (a), 0.50 mL of reference solution (b) and 10.0 mL
Test solution Dissolve 25.0 mg of the substance to be of the internal standard solution to 100.0 mL with methylene
examined in the mobile phase and dilute to 100.0 mL with chloride R.
the mobile phase. . Golumn:
Reference solution Dissolve 25.0 mg of 4-nitrobenzak acid R - material: fused silica,
and 25.0 mg of benzocaine R in methanol R and dilute to - size: 1= 30 m, 0 ;;;; 0.32 mm,
100.0 mL with the same solvent. Dilute 1.0 mL to 50.0 mL - stationary phase: phenyl(5)m,thyl(95)poIysiloxane R (film
with the mobile phase. Dilute 1.0 rnL of this solution to thickness 0.5 pm).
10.0 mL with the mobile phase. Carrier gas helium for chromatography R.
Column: Flow rate 1.0 mUmin.
- size: 1= 0.12 m, 0 = 4.0 mm, Split ratio 1:10.
- stationary phase: oaylsilylsilica gelfor ehronuuography R
Temperature:
(5 pm).
"'Iobile phase Mix 20 volumes of a mixture of 70 volumesof Time Temperature
acetonitrile R and 80 volumes of methanol R) and 80 volumes (min) CCJ
of a solution containing 1.5 gIL of potassium dihydrogen Column 0-4 130
phosphate Rand 2.5 gIL of sodium octanesulfonate R adjusted 4 - 6.5 J30 --> 180
to pH 2.2 with phosplwric acid R. 6.5 - 11.5 lBO
Flow rate 1.0 mUmin. Injection port 2BO
Detection Spectrophotometer at 270 run. Detector 300
Injection 20 pL.
Run time II times the retention time of 4-aminobenzoic Detection Flame ionisation.
acid. Injection 2 ~L; inject the test solution and reference
Relative retention With reference to 4-aminobenzoic acid solution (c).
(retention time = about 3 min): impurity A = about 4; Retention lime Internal standard = about9.5 min.
impurity B = about 9. Limits:
Limits: - impurity G: calcuiate the ratio (R) of the area of the peak
- impuniy A: not more than the area of the corresponding due to impurity C to the area of the peakdue to the
peak in the chromatogram obtained with the reference internal standard from the chromatogram obtained with
solution (0.2 per cent» reference solution (c); calculate the ratio of the area of the
- impuniy B: not more than the area of the corresponding peak due to impurity C to the area of the peak due to the
peak in the chromatogram obtained with the reference internal standard from the chromatogram obtained with
solution (0.2 per cent) the test solution: this ratiois not greater than R (10 ppm),
- any other impurity: not more than 0.5 times the area of the - impurity D: calcuiate the ratio (R) of the area of the peak
peak due to impurity A in the chromatogram obtained due to impurity D to the area of the peak due to the
with the reference solution (0.1 per cent), internal standard from the chromatogram obtained with
- total: not more than 2.5 times the area of the peak due to reference solution (c)j calculate the ratio of the area of the
impurity A in the chromatogram obtainedwith the peak due to impurity D to the area of the peak due to the
reference solution (0.5 per cent), internal standard from the chromatogram obtained with
- disregard limit: 0.1 times the area of the peak due to the test solution: this ratio is not greater than R (10 ppm).
impurity A in the chromatogram obtained with the Iron (2.4.9)
reference solution (0.02 per cent). Maximum 40 ppm.
Impurity C and impurity D Dissolve 0.250 g in 3 mL of ethanol (96 per cen!! Rand
Gas chromatography (2.2.28). dilute to 10.0 mL with waterR.
Internal standard solution Dissolve 20.0 mg of lauric acid R in Water (2.5.12)
methylene chloride R and dilute to 100.0 mL with the same Maximum 0.2 per cent, determined on 1.00 g.
solvent.
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1-148 Aminocaproic Acid 2022
Sulfated ash (2.4.14) A. Examine by infrared absorption spectrophotometry

Maximum 0.1 per cent, determined on 1.0 g. (2.2.24), comparing with the spectrum obtained with
ASSAY ommocaproic acid CRS. Examine the substances prepared as
discs.
Dissolve 0.100 g with heating in 50 mL of carbon dioxide-free
water R. Titrate with 0.1 M sodium hydroxide determining the B. Examine the chromatograms obtained in the test for
end-point potentiometrically (2.2.20). ninhydrin-positive substances. The principal spot in the
chromatogram obtained with the test solution (b) is similar in
1 mL of 0.1 M sodium hydroxide is equivalent to 13.71 mg of
position,colour and size to the principal spot in the
C,H,NO,.
chromatogram obtained with reference solution (a).
STORAGE C. Dissolve 0.5 g in 4 mL of a mixture of equal volumes of
Protected from light. '0
diJu« hydrochloric acid Rand warer R. Evaporate dryness by
IMPURITIES heatingon a water-bath. Dry the residue in a desiccator.
Dissolve the residue in about 2 mL of boiling ethanol R.
Allow to cool and maintain at 4 °C to 8 °C for 3 h. Filter
under reduced pressure. The residue washed with about
10 mL of acetone R and dried a' 60°C for 30 min, melts
(2.2.14) at 131°C to 133°C.
A. 4-nirrobenzoic acid, D. Dissolve about 5 mg in 0.5 mL of distiJled water R.
Add 3 mL of dimethylfrmnamide Rand 2 mL of ascorbic acid
solution R. Heat on a water-bath. An orange colourdevelops.
TESTS
Solution S
Dissolve 10.0 g in carbon dioxide-free water Rand dilute '0
B. ethyl 4-aminobenzoate (benzocaine), 50.0 mL with the same solvent.
Solution S is colourless (2.2.2, Method If) and remains clear
(2.2.1) on standing for 24 h.
pH (2.2.3)
The pH of solution S is 7.5 10 8.0.
C. aniline,
Absorbance (2.2.25)
A. The absorbance of solution S at 287 run is not more
than 0.10 and al450 om is not more than 0.03.
B. Place 2.0 g in an even layer in a shallow dish 9 em in
diameter, coverand allow to stand at 98°C to 102°C for
D. 4-methylaniline (fr-,oluidine). 72 h. Dissolve in water R and dilute to 10.0 mL with the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PO Ell same solvent. The absorbance of the solution at 287 run is
not more than 0.15 and at 450 run is not more than 0.03.
Ninhydrin-positive substances
Aminocaproic Acid suitable silica gel as the coating substance.
(ph. Eur. monograph 0874) examined in water R and dilute to 10 mL with the same
solvent.
Test solUlion (b) Dilute 1 mL of test solution (a) to 50 mL
with wattr R.
131.2 6()'32-2
Reference solution (a) Dissolve 10 mg of aminocaproic
Action and use '0
acid CRS in water R and dilute 50 mL with the same
Antifibrinolytic. solvent.
POEII _
Reference SolUlion (b) Dilute 5 mL of test solution (b) '0
20 mL with waterR.
DEFINITION Reference solution (e) Dissolve 10 mg of ominocaproic
Aminocaproic acid contains not less than98.5 per cent and acidCRS and 10 mg of leucine CRS in water Rand dilute '0
not more than the equivalent of 101.0 per cent of 25 mL with the same solvent.
6-aminohexanoic acid, calculated with reference to the dried '0
Apply separately the plate 5 flL of each solution. Allow the
substance. plate to dry in air. Develop over a path'of 15 em using a
CHARACTERS mixture of 20 volumes of glacio! acetic acid R, 20 volumesof
A white or almost white, crystalline powder or colourless water Rand 60 volumes of butanol R. DIY the plate in a
crystals, freely soluble in water, slightly soluble in alcohol. current of warm air. Spray with ninhydrin solution R and heat
It melts at about 205 "C with decomposition.
at 100°C '0 105°C for 15 min. Any spot in the
chromatogram obtained with the test solution (a), apart from
IDENTIFICATION me principal spot, is not more intense than the spot in the
Fi", identification: A. chromatogram obtained withreference solution (b)
Second identification: B, C, D. (0.5 per cent). The test is not valid unless the chromatogram
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2022 Aminoglutethimide 1-149
obtained with referencesolution (c) shows two clearly lvlobiJe phase glacialacetic acid R, methanol R, ethyl acetate R
separated principal spots. (0.5: 15:85 VIV/lI).
Loss on drying (2.2.32) Application 5 ~L.
Not more than 0.5 per cent, determined on 1.000 g by Development Over 3/4 of the plate.
drying in an oven at 105°C. Drying In air.
Sulfated ash (2.4.14) Detection Examine in ultraviolet light at 254 om.
Not more than 0.1 per cent, determined on 1.0 g.
ASSAY - the chromatogram shows 2 clearly separed spots.
Dissolve 0.100 g in 20 mL of anhydrous acetic acidR. Using Results The principal spot in the chromatogram obtained
0.1 mL of crystal violel solution R as indicator) titrate with with me test solutionis similar in position and size to the
0.1 M perchloric acid until the colour changes from bluish- principal spot in the chromatogram obtained with reference
violet to bluish-green. solution (a).
1 mL of 0.1 M perchloric acid is equivalent to 13.12 mg of TESTS
C6H"N02 • Solution S
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE"
Dissolve 1.0 g in mahand R and dilute (0 20.0 mL with the
same solvent.
Aminoglutethimide than reference solution Y7 (2.2.2" Method If).
(Ph. Eur. monograph 1291) Optical rotation (2.2.7)
--0.10° to + 0.10°" determined on solution S.
Related substances
and enanUomer
So/vent mixture methanol R, acetate buffer solution pH 5.0 R
(50:50 VW).
examined in me solvent mixture and dilute to 50.0 mL with
232.3 125-84-8
me solvent mixture.
Acdon and use Reference solution (aJ Dissolve 5.0 mg of aminoglutethimide
Inhibitor of adrenal corticosteroid synthesis; used in chemical impurity A CRS in the solvent mixture and dilute to 25.0 mL
adrenalectomy. with the solvent mixture.
PIlE" _
to 10.0 mL with me solvent mixture.
DEFINITION Reference solution (c) Dilute 1.0 mL of the test solution to
(3RSJ-3-(4-Aminophenyl)-3-ethylpiperidine-2,6-dione. 100.0 mL with the solventmixture.
Content Reference solution (d) Dilute 1.0 mL of the test solution to
98.0 per cent to 101.5 per cent (dried substance). 10.0 mL with reference solution (a).
CHARACTERS Column:
Appearance - size: 1= 0.15 m, 0 = 3.9 mm;
White or slightly yellow, crystalline powder. - stationary phase: octadecylsilyl silica gd for chromatography R
(4 urn);
Solubility - temperature: 40°C.
Mobile phase Mix 27 volumes of methanol Rand 73 volumes
soluble in methanol.
of acetate bll!fer solution pH 5.0 R.
IDENTIFICATION Fkno rate ·1.3 mUmin.
Injection 10 J1L of me test solution and reference
A. Melting point (2.2.14): 150°C to 154 °C. solutions (b), (c) and «1).
B. Infrared absorption spectrophotometry (2.2.24). Run time 4 times the retention time of aminoglutethimide.
Comparison ominoglmethimide CRS. Identification of impuniies Use the chromatograrn obtained
C. Thin-layer chromatography (2.2.27). with reference solution (b) to identify the peak due to
Test solution Dissolve 25 mg of the substance to be impurity A.
examined in acetone R and dilute to 5 mL with the same Relative retention Withreference to arninoglutethimide
solvent. (retention time =about 9 min): impurity A = about 1.3.
Reference solution (a) Dissolve 25 mg of System suiU1.bility Reference solution (d):
aminoglutelhimide CRS in aurone R and dilute to 5 mL with - resohuion: minimum 2.0 between me peaks due to
the same solvent. aminoglutethimide and impurity A.
Reference solution (b) Dissolve 25 mg of Limits:
aminoglutethimide CRS and 25 mg of glutethimide CRS in - impurityA: not more man twice the area of the principal
aUlane R and dilute to 5 mL with the same solvent. peak in me chromatogram obtained with reference
Plate TLC silica gd F254 ptate R. solution (b) (2.0 per cent);
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1-150 Aminophylline 2022
- unspecified impurities: for each impurity, not more than in the monograph. They are limited by the general acceptance
0.1 times the area of the principal peak in the criterion for other/unspecified impurities and/or by thegeneral
chromatogram obtained with reference solution (e) mrmograph Substances for pharmaceutical use (2034). It is
(0.10 per cent); therefore not neussaryto identify these impumies for
- sum of impunties other than A: not more than the area of demonstration of compliance. See also 5.10. Control of impurities
the principal peak in the chromatogram obtained with in substances for pharmaceutical use) B, C.
reference solution (e) (1.0 per cent);
- total: maximum 2.0 per cent for the sum of the contents
of aU impurities;
- disregard limit: 0.05 times the area of the principal peak in andenantiomer
(0.05 per cent).
Impurity D
liquid chromatography (2.2.29). Carryom the tesr prouaed A. (3RS)-3-(3-aminophenyl)-3-ethylpiperidine-2,6-dione
from light. Use shaking, not sonication or heat, to dissolve the (3-aminoglutethimide),
reference substance and thesubstance to be examined.
examined in dimethyl sulfoxide R and dilute to 100.0 mL with
andenantiomer
the same solvent.
Reference solution Dissolve 3.0 mg of aminoglutethimide
impurity D CRS in dimethyl sulfoxide R and dilute to
100.0 mL with the same solvent. Dilute 1.0 mL of this
B. (3RS)-3-ethyl-3-(3-nitrophenyl)piperidine-2,6-dione,
solution to 100.0 mL with dimethyl sulfoxide R.
Column:
- size: I:::: 0.12 ffi, 0:::: 4 mm; 0y~yO~
- stationary phase: o<tadecy/si(y/ siliw gelfor chromatography R ~N02 and enanliomer
(5 urn). >-
Mobile phase Dissolve 0.285 g of sodium edetate R in H,C
water R, add 7.5 mL of dilute acetic acid Rand 50 mL of
0.1 M potassium hydroxide and dilute to 1000 mL with C. (3RS)-3-ethyl-3-(4-nitrophenyl)piperidine-2,6-dione,
water R; adjust to pH 5.0 withglacial acetic acid R; mix
350 mL of this solution with 650 mL of methanol R.
Flow rate 1.0 mIJmin.
Injection 10 ~L.
Sysrem suitability Test solution:
- number of theoretical plates: minimum 3300, calculated for
the principal peak; D. 3,3'-[diazenediylbis(4.I-phenylene))bis(3-ethylpiperidine-
- massdistribution ratio: 2.0 to 5.0 for the principal peak; 2,6-dione) (azoglutethimide).
- symmetry factor: maximum 1.2 for the principal peak. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
Limit:
- impurity D: not more than the area of the principal peak
in the chromatogram obtained with thereference solution
(300 ppm).
Aminophylline ***
Sulfates (2.4.13) *** ***
Maximum 500 ppm. (TheophyUine-Ethy/enediamine, Ph. Eur. mrmograph ***
Dilute 6 mL of solution S to 15 mL with distilled water R. 0300)
an oven at 105 °C"
Maximum 0.1 per cent, determined on 1.0 g-
ASSAY
Dissolve 0.180 g in 50 mL of anhydrous acetic acid Rand
titrate with 0.1 M perchlon"c acid, determining the end-point 420.4 317-34-0
potentiometrically (2.2.20).
Action and use
1 mL of O. I M perchknU acid is equivalent to 23.23 mg Non-selective phosphod.iesterase inhibitor; treatment of
of C 13H,oN,O,. reversible airways obstruction.
IMPURITIES Preparations
Specified impurities A, D. Aminophylline Injection
Other detectable impurities (thefol1m»ing substances would, if Aminophylline Tablets
present at a sufficient level, be detected by one or other of the tests Aminophylline Prolonged-release Tablets
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2022 Aminophylline 1-151
PhEIl _ solution and dilute to 100 mL with the mobile phase. Dilute
DEFlNmON 5 mL of this solution to 50 mL with the mobile phase.
Content Column:
- theophylline (C 7H,N,O,; M, 180.2): 84.0 per cent to - size: / = 0.25 m, 0 == 4 mm;
87.4 per cent. (anhydrous substance); - stationary phase: octodecylsilyl silica gelfor chromategraphy R
- elhylenediamine (C,H.N,; M, 60.1): 13.5 per cent to (7 urn).
15.0 per cent (anhydrous substance). Mobile phase Mix 7 volumes of acetonitrile for
chromatography Rand 93 volumes of a 1.36 gIL solution of
CHARACTERS
sodium acetate R containing 0.50 per cent. VIV of glacial acetic
Appearance
acidR.
White or slightly yellowish powder, sometimes granular,
hygroscopic. Flow rate 2.0 mUmin.
Solubility Daeaion Spectrophotometer at 272 nm.

Freelysoluble in water (the solution becomes cloudy through Inj«tion 20 ~L.
absorption of carbon dioxide), practically insoluble in Run lime 3.5 times the retention time of theophylline.
anhydrous ethanol. Relative retention With reference to theophylline (retention
IDENTIFICATION time = about 6 min): impurity G = about 0.6.
First identification: B~ C, E. System suitability Reference solution (b):
Second identification: A, C, D, E, F. - resolution: minimum 2.0 between the peaksdue to
impurity G and theophylline.
Dissolve 1.0 g in 10 mL of waterR and add 2 mL of dilute
hydro<hlol"ic acidR dropwise with shaking. Filter. Use the Limits:
precipitate for identification tests A, B, D and F and the - unspuified impun"u"eJ: for each impurity, not more than the
filtrate for identification test C. area of the principal peak in the chromatogram obtained
with referencesolution (a) (0.10 per cent),
A. Melting point (2.2./4): 270"C to 274 "C, determined
after washing the precipitate with water R and dryingat
105 "C.
(0.10 per cent);
B. Infrared absorption spectrophotometry (2.2.24). - disregard Iimit: 0.5 times the area of the principal peak in
Preparation Precipitate, washed with water R and dried at the chromatogram obtained with reference solution (a)
105 "C. (0.05 per cent).
Comparison theophyOine CRS. Water (2.5./2)
C. To the filtrate add 0.2 mL of benzoylchloride R, make Maximum 1.5 per cent, determined on 0.50 g.
alkaline with diJu'" sodium hydroxide solwion R and shake Sulfated ash (2.4./4)
vigorously. Filter the precipitate, wash with 10 mL of Maximum 0.1 per cent, determined on 1.0 g.
waterR, dissolve in 5 mL of hor ethanol (96 per cenQ Rand
add 5 mL of water R. A precipitate is formed, which, when ASSAY
washed and dried at 105 "C, melts (2.2./4) at 248 "C to Ethylenediamine
252 "C. Dissolve 0.250 g in 30 mL of waUll" R. Add 0.1 mL of
bromocresol green solution R. Titrate with 0.1 M hydro<hlOl"ic
D. Heat about 10 mg of the precipitate with 1.0 mL of a
acid until a green colour is obtained.
360 gIL solution of porassium hydroxide R in a water-bath at
90 "C for 3 min, then add 1.0 mL of diazotised suljoniJic acid I mL of O. / M hydrochloric acid is equivalent to 3.005 mg of
solution R. A red colourslowly develops. Carry out a blank C,H.N,.
test. Theophy1llne
E. Water (see Tests). Heat 0.200 g to constant mass in an oven at 135 "C.
F. The precipitate gives the reaction of xanthines (2.3.1). Dissolve the residue with heating in 100 mL of water R)
allow to cool) add 20 mL of 0.1 M silvernitrate and shake.
TESTS Add I mL of bromothymol bluesolution R/. Titrare with O. / M
Appearance of solution sodium hydroxide.
The solution is not more opalescent than reference I mL of 0./ M sadium hydroxide is equivalent to 18.02 mg of
suspension Il (2.2.1) and not more intensely coloured than C 7HsN402 "
reference solution GY. (2.2.2, Merhad If).
STORAGE
Dissolve 0.5 g with gentle warming in 10 mL of carbon
dioxide-free waUll" R. In an airtight container) protected from light.
Related substances IMPURITIES
Liquid chromatography (2.2.29). Otherdetectable impurities (thefollowing substances would, if
Test solution Dissolve 47 mg of the substance to be present at a sufficientlevel be detected by oneor other of the lMts
J
examined in the mobile phase and dilute to 20.0 mL with in the monograph. They aw limited by the general a«eptame
the mobile phase. criterion for otherhmspedfied impurities and/or by thegeneral
monograph Substances for phannaceutical us, (2034). It is
Reference solution (a) Dilute 1.0 mL of the test solution to therefore not neassary to identify these impun"ties for
100.0 mL with the mobile phase. Dilute 1.0 mL of this demonstration of compliance. See also 5"10. Conrrol of impun",ies
solution to 10.0 mL with the mobile phase. in substances for pharmaceutical use) A) B) CJ DJ EJ FJ G.
Reference solution (b) Dissolve 10 mg of theobromine R
(impurity G) in the mobile phase, add 5 mL of the lest
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1-152 Aminophylline Hydrate 2022
Aminophylline Hydrate
(TheophyUine-Ethyknediamine Hydrare, Ph. Eur.
monograph 0301)
A. 1,3,7-trimethyl-3,7-dihydro-IH-purine-2,6-dione
(caffeine),
.""'0
C,.H,,N,oO,,xH,O 420.4 72487-55-9

(anhydrous substance)
B. 3-methyl-3,7-dihydro-IH-purine-2,6-dione, Action and use

Non-selective phosphodiesterase inhibitor; treatment of
reversible airways obstruction.
Preparations
Aminophylline Injection
Aminophylline Tablets
Amin~~llineProron~d~ere~eTookts
C. N-(6-amino-l,3-dimemyl-2,4-dioxo-l,2,3,4- PhE" _
tetrahydropyrimidin-5-yl)fonnamide,
DEFINITION
Content
H,C'N~N - meophyUine (C,H"N,O,; M, 180.2): 84.0 per cent to
87.4 per cent (anhydrous substance);
H)l)
HN N - ethylenediamine (C,HaN,; M, 60.1): 13.5 per cent to
, H 15.0 per cent (anhydrous substance).
. CH,
D. N-methyl-5-(methylamino)-IH-imidazole-4-carboxamide, CHARACTERS
Appearance
Whiteor slightly yellowish powder, sometimes granular.
Solubl1lty
Freely soluble in water (the solution becomes cloudy through
absorption of carbon dioxide), practically insoluble in
anhydrous ethanol.
E. I,3-dimethyl-7,9-dihydro-IH-purine-2,6,8(3H)-trione, IDENfIFICATION
First ;dentifkation: B. C, E.
Second identification: A, C, D, E, F.
Dissolve 1.0 g in 10 mL of water R and add 2 mL of dl7ute
hydro<hlori< acid R dropwise with shaking. Filter. Use the
precipitate for identification testsA, B, D andF and the
filtrate for identification test C.
A. Meiring point (2.2.14): 270 -c to 274 -c, determined
F. 7-(2-hydroxyethyl)-1,3-dimethyl-3,7-dihydro-IH-purine- after washing the precipitate with water R and drying at
2,6-dione (erofyUine), 105°C.
Preparation Precipitate, washed with water R and dried at
105°C.
Comparison meophyUine CRS.
C. To the liltrate add 0.2 mL of benzoyl chloride R, make
alkaline with dilure sodium hydroxide sduuon R and shake
G. 3,7-<1imethyl-3,7-<1ihydro-IH-purine-2,6-<1ione vigorously. Filterthe precipitate, washwith 10 mL of
(theobromine). water R, dissolve in 5 mL of hot ethano/ (96 per cent) R and
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" add 5 mL of water R. A precipitate is formed which, when
washed and dried at 105 "C, melts (2.2.14) at 248 "C to
252°C.
D. Heat about 10 mg of the precipitate with 1.0 mL of a
360 gIL solution of potassium hydroxide R in a water-bath at
90 "C for 3 min, then add 1.0 mL of diazotised sulfanilic acid
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2022 Aminophylline Hydrate 1-153
solution R. A red colour slowly develops. Carry out a blank Theophylline

test. Heat 0.200 g to constant mass in an oven at 135 "C.
E. Water (see Tests). Dissolve the residue with heating in 100 mL of water RJ
F. The precipitate gives the reaction of xanthines (2.3.1). allow to ccol, add 20 mL of 0.1 M silver nitrate and shake.
Add I mL of bromolhymol blee solution Rt. Titrate with 0.1 M
TESTS sodium hydroxide.
Appearance of solution I mL of 0.1 M sodium hydroxide is equivalent to 18.02 mg of
The solution is not more opalescent than reference C,HgN.O,.
suspension IT (2.2.1) and not more intensely coloured than
reference solution GY. (2.2.2, Method II). STORAGE
Dissolve 0.5 g with gentle wanning in 10 mL of carbon In a well-filled, airtight container, protected from light.
dioxide-free warer R. IMPURITIES
Related substances Otherdetectable impumies (thefol/qwjng substances would, if
Liquid chromatography (2.2.29). present at a sufficient Ieuel, be deucud by one or other 0/ the tests
Test solution Dissolve 50 mg of the substance to be in the monograph. They are lJ"miud by thegeneral acceptance
examined in the mobile phase and dilute to 20.0 mL with critenon for other/unspecified impurities and/or by the general
the mobile phase. monograph Substances for pharmaceutical use (2034). 1, is
demonstration 0/compliance. See also5.10. Control oj impurities
in substances Jarpharmaceutical use) A J BJ CJ D J EJ F, G.
Reference solution (b) Dissolve 10 mg of theobromine R
(impurity G) in the mobile phase, add 5 mL of the test
solution and dilute to 100 mL with the mobile phase. Dilute
5 mL of this solution to 50 mL with the mobile phase.
Column:
- size: 1= 0.25 m, «2) = 4 mm;
- stationary phase: octade<y/silyl silica gelfor chromatography R
A. 1,3,7-bimethyl-3,7-dihydro-IH-purine-2,6-dione
(7 urn).
(caffeine),
MoMe phase Mix 7 volumes of acetonitTile for
chromatography Rand 93 volumes of a 1.36 gIL solution of
sodium autate R containing 0.50 per cent V/V of gladal acetic
acidR.
Injection 20 ~L.
Run time 3.5 times the retention time of theophylline. B. 3-methyl-3,7-dihydro-IH-purine-2,6-dione,
Re1au"ve retention With reference to theophylline (retention
= =
time about 6 min): impurity G about 0.6.
System suitabiHt;y Reference solution (b):
- resolution: minimum 2.0 between the peaksdue to
impurity G and theophylline.
Limits:
area of the principal peak in the chromatogram obtained C. N-(6-amino-I,3-dimemyl-2,4-dioxo-l J2,3,4-
with reference solution (a) (0.10 per cent); tetrahydropyrimidin-5-yl)formantide,
- total: not more than me area of the principal peakin the
o
(0.1 per cent); H,C, ~N
- disregard Iimit: 0.5 times the area of the principal peak in
~ HN
. . J-N>
I H
(0.05 per cent). CH,
Water (2.5.12)
3.0 per cent to 8.0 per cent, determined on 0.50 g. D. N-methyl-5-(methylamino)-IH-imidazole-4-ClIrboxamide,
Sulfated ash (2.4.1f)
ASSAY
Ethylenediamine
Dissolve 0.250 g in 30 mL of warer R. Add 0.1 mL of
bromocresol green solution R. Titrate with 0.1 M hydrochlori<
acid until a green colour is obtained.
E. 1,3-dimethyl-7,9-dihydro-IH-purine-2,6,8(3H)-trione,
I mL of 0.1 M hydrochloric acid is equivalent to 3.005 mg of
C,H.N,.
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1-154 Amiodarone Hydrochloride 2022
pH (2.2.3)
3.2 to 3.8.
Dissolve 1.0 g in carbon dioxide-free water R, heating at 80°C,
cool and dilute to 20 mL with the same solvent.
ImpurltyH
Thin-layer chromatography (2.2.27). Prepare the solurions
F. 7-(2-hydroxyethyl)-1 ,3-dimethyl-3,7-dihydro-I H-purine-
immediately before use and keep protected from bright light
2,6-dione (etofylline),
examined in methylene chloride R and dilute to 5.0 mL with
the samesolvent.
Reference solurion (a) Dissolve 10.0 mg of (2-chloroethyl)
drethy/amine hydrochloride R (impurity H) in methylene
chloride R and dilute to 50.0 mL with the same solvent.
Dilute 2.0 mL of the solution to 20.0 mL with methylene
G. 3,7-dimethyl-3,7-dihydro-IH-purine-2,6-dione chloride R.
(theobromine). Reference solurion (b) Mix 2.0 mL of the lest solution and
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII 2.0 mL of reference solution (a).
Plate TLC silica gelF,,. plate R.
Mobile phase anhydrous formic acidR, methanol R, methylene
chloride R (5: 10:85 VIVIV).
Amiodarone Hydrochloride ***
** ** Application 50 J.lL of me test solutionand reference
solution (a); 100 ~L of reference solution (b).
(Ph. Eur. mmograph 0803) *****
H3C o Drying In a current of cold air.
Detection Spray with potassium iodobismuthate solurion Rl and
then with dilute hydrogen peroxide solution R; examine
immediately in daylight.
- the spot due to impurity H is clearly visible.
682 19774-82-4 Limit.
- impuniy H: any spot due to impurity H is not more
Action and use intense than the spot in the chromatogram obtained with
Potassium channel blocker; class m antiarrhythmic. reference solution (a) (0.02 per cent).
Preparations Related substances
Amiodarone Infusion Liquid chromatography (2.2.29).
Amiodarone Oral Suspension Buffersolurian pH 4.9 To 800 mL of water R add 3.0 mL of
Amiodarone Tablets glacial acetic acidR, adjust to pH 4.9 with dilute ammonia Rl
PIIEII _ and dilute to 1000 mL with water R.
Test solurion Dissolve 0.125 g of the substance to be
DEFINITION examined in a mixture of equal volumes of acewnitrile Rand
(2-Butylbenzofuran-3-yl)[4- [2-(diethylamino)ethoxy)-3,5- water R and dilute to 25.0 mL with the same mixture of
diiodophenyljmethanone hydrochloride. solvents.
Content Referenu solution Dissolve 5 mg of amiodarone
98.5 per cent to 101.0 per cent (dried substance). impurity D CRS, 5 mg of amiodarone impurity E CRS and
CHARACTERS 5.0 mg of amiodarone hydrochloride CRS in methanol Rand
Appearance
dilute to 25.0 mL with the same solvent. Dilute 1.0 mL of
the solution to 20.0 mL with a mixture of equal volumes of
White or almost white,fine) crystalline powder.
acetoninfle R and water R.
Solublllty
Column:
Very slightly soluble in water, freely soluble in methylene
- size: / = 0.15 m, 0 = 4.6 mm;
chloride, soluble in methanol, sparingly soluble in ethanol
- stationary phase: end-<apped octad«Y1si1y1 silica gelfor
(96 per cent).
chromatography R (5 ~);
IDENTIFICATION - temperature: 30 °C.
A. Infrared absorption spectrophotometry (2.2.24). Mobile phase Buffer solution pH 4.9, methanol R,
Comparison amiodarone hydrochloride CRS. acetonitrile R (30:30:40 VIVIV).
B. It gives reaction (b) of chlorides (2.3.1). Flow rate I mUmin.
TESTS Detection Spectrophotometer at 240 om.
Appearance of solution Injection 10 flL.
The solution is clear (2.2.1) and not more intensely coloured Run time Twice the retention time of amiodarone.
than reference solution GY, or BY, (2.2.2, Method If). Relative retention With reference to amiodarone (retention
Dissolve 1.0 g in methanol R and dilute to 20 mL with the time =about 24 min): impurity A =about 0.26;
same solvent.
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2022 Amiodarone Hydrochloride 1-155
o andeoentcrner
impurity D = about 0.29; impurity E = about 0.37; H,C
impurity B = about 0.49; impurity C = about 0.55;
= =
impurity G about 0.62; impurity F about 0.69.
impurities D and E.
Limits: A. (2-butylbenzofuran-3-yl)[4-[2-(diethylamino)ethoxy]
- impuniies A, B, C, D, E, F, G: for each impurity, not phenyl]methanone,
more than the area of the peakdue to amiodarone in the
chromatogram obtained with the reference solution andenantiomer
H,C
(0.2 per cent);
- unspecified impunues: for eachimpurity, not more than
0.5 times the area of the peak due to amiodarone in the
chromatogram obtained with me reference solution
(0.10 per cent);
- total: not more than 2.5 times the area of the peak due to
amiodarone in the chromatogram obtained with the B. (2-butylbenzofuran-3-yl)[4-[2-(ethylamino)ethoxy]-3,5-
reference solution (0.5 per cent); diiodophenyl]methanone,
- disregard limit: 0.25 times the area of the peak due to and enensomer
amiodarone in the chromatogram obtained with the H,C o
reference solution (0.05 per cent).
Iodides
Maximum 150 ppm.
Prepare the tesland reference solutions simultaneously,
Solution A Add 1.50 g of the substance to be examined to
40 mL of water R at 80°C and shake until completely C. (2-butylbenzofuran-3-yl)[4-[2-(diethylamino) ethoxy]-3-
iodophenyJ]methanone,
dissolved. Cool and dilute to 50.0 mL with water R.
Test solution To 15.0 mL of solution A add 1.0 mL of H,C o
0.1 M hydrochloric acid and 1.0 mL of 0.05 M potassium
iodate. Dilute to 20.0 mL with woW R. Allow to stand
protected from light for 4 h.
OH
Reference solution To 15.0 mL of solution A add 1.0 mL of
0.1 M hydrochloric acid, 1.0 mL of an 88.2 mgIL solution of
potassium iodide Rand 1.0 mL of 0.05 M potassium iodate.
D. (2-butylbenzofuran-3-yl)(4-hydroxy-3,5-
Dilute to 20.0 mL with water R. Allow to stand protected
diiodophenyl)methanone,
from light for 4 h.
Measure the absorbances (2.2.25} of the solutions at 420 nm, H,C o
using a mixtureof 15.0 mL of solution A and 1.0 mL of
0.1 M hydrochloric acid diluted to 20.0 mL wirb waW R as
the compensation liquid. The absorbance of the test solution OH
is not greater than half the absorbance of the reference
solution.
Loss on drying (2.2.32) E. (2-butylbenzofuran-3-yJ)(4-hydroxyphenyl)methanone,
Maximum 0.5 pet cent, determined on 1.000 g by drying at
50°C at a pressure not exceeding 0.3 kPa for 4 h. H,C
OH
ASSAY
Dissolve 0.600 g in a mixture of 5.0 mL of
0.01 M hydrochloric add and 75 mL of ethanol (96 per cent) R.
Carry out a potentiometric titration (2.2.20), using F. (2-butylbenzofuran-3-yl)(4-hydroxy-3-iodophenyl)
0.1 M sodium hydroxide. Read the volume added between the methanone,
2 points of inflexion.
I mL of 0.1 M sodium hydroxide is equivalent to 68.18 rng of
C 2,H,oC1I2NO,. andenantlomer
STORAGE
Protected from light, at a temperature not exceeding 30°C.
IMPURITIES
Specified impurities A, B, C, D, E, F, G, H.
G. [4-[2-(diethylamino)ethoxy]-3,5-diiodophenyl] [2-[(IRS)-
l-methoxybutyl]benzofuran-3-yl]methanone,
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1-156 Amisulpride 2022
Pkue TLC silica gel G plate R.

Mobile phase 50 per cent VIV solutionof concentrated
ammonia R, anhydrous ethanol R, di'-isupropyl ether R
(10:25:65 VIVIV); use the upper layer obtained after shaking
H. 2-chloro-N,N-diethylelhanamine (2-chlorotriethylamine,
the mixture.
(2-cWoroethyl)diethylamine).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" Application I0 ~L.
Deodopmenc Over 213 of the plate.
Drying In air.
Detection Spraywith ninhydrin sotudon R and heat at
Amisulpride *** 100-105 °C for 15 min.
*** *** Retardation factors Impurity A = about 0.2j
(Ph. Eur. monograph 1490) *** amisulpride = about 0.5.
System suitability The chromatogram obtained with
reference solution (b) shows 2 clearly separated spots.
andenantiomer Limit:
- impurityA: any spot due to impurity A is not more
intense than the corresponding spot in the chromatogram
obtainedwith reference solution (a) (0.1 per cent).
369.5 71675-85-9 Related substances
Action and use
Solvent mixture acetomiri/e R, methanol R, mobile phase A
Dopamine receptor antagonist; neuroleptic.
(12:16:72 VIVIV).
Preparations Test solution Dissolve 0.100 g of the substance to be
Amisulpride Oral Solution examined in 16 rnL of methanol R, add 12 rnL of
Amisulpride Tablets acetonitrile R and dilute to 100.0 mL with mobile phase A.
PhE" _ Reference solution (a) Dilute 1.0 rnL of the test solution to
100.0 mL with the solvent mixture. Dilute 1.0 rnL of this
DEFINlTION solution to 10.0 mL with the solvent mixture.
4-Amino-N-[[(2RS)-I-ethylpyrrolidin-2-yl]methyl]-5-
Reference solution (b) Dissolvethe contentsof a vialof
(ethylsulfonyl)-2-methoxybenzamide.
amisu/pride for system suilability CRS (containing impurity B)
Content in 1 mL of the solvent mixture.
99.0 per cent to 101.0 per cent (dried substance). Column:
CHARACTERS - size: 1 =-0.25 m, 0 = 4.6 nun;
Appearance - s,",wnary phase: base-deactivated oayIsiIyl silica gelfor
White or almostwhite, crystalline powder. chromatography R (5 1"");
Solubility
Practically insoluble in water, freely soluble in methylene Mobile phase:
chloride) sparingly soluble in anhydrous ethanol. - mobile phase A: dissolve 0.7 g of sodium """'nesulfonate R in
930 rnL of waterfor chromalOgraphy R and add 45.0 rnL
mp
of a 5 per cent VIV solution of dilute sulfwU acidR; adjust
About 126 °C.
to pH 2.3 with dilute sulfuric acid R and dilute to
IDENTIFICATION 1000 rnL with waterfor chromatography R;
Infrared absorption spectrophotometry (2.2.24). - mobile phase B: methanol Rl ;
Comparison amisulpride CRS. - mobile phase C: aatonitrile for chromatography R;
TESTS
Tbne Mobile phase A Mobile phase B Moblle phase C
Appearance of solution (min) (per cent VIP) (per cent VII') (per cent VII')
The solution is clear (2.2.1) and not more intensely coloured 0-18 72 I. 12
than reference solution Y6 (2.2.2J Method If). 18 - 35 72 ---> 50 16 ---> 38 12
Dissolve 1.0 g in 3 mL of a mixture of 1 volumeof auae
acid Rand 4 volumes of water R, and dilute to 20 rnL with Flow ral< 1.5 mllmin.
water R.
Detection Spectrophotometer at 225 nm,
Impurity A
Injection 10 J1l..
Identification 0/impurities Use the chromatogram obtained
with reference solution (b) to identify the peakdue to
impurity B.
solvent.
Relative retention With reference [0 amisulpride (retention
Reference solurian (a) Dissolve 5 mg of sulpiride
impuniy A CRS (amisulpride impurity A) in methanol Rand
= =
dilute to 25 mL with the same solvent. Dilute 2 mL of the System suitability Reference solution (b):
solution to 20 mL with methanol R. - peak-w-vaJky ratio: minimum 2.0, where Hp = height
above the baseline of the peakdue to impurity Band
Reference solution (b) Dilute I rnL of the test solution to
H; = height above the baseline of the lowest point of the
10 mL with reference solution (a).
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2022 Amitriptyline Hydrochloride 1-157
curve separating this peak from the peak due to

amisulpride.
- for each impurity, use the concentration of amisulpride in
E. 4-amino-5-(ethylsulfonyl)-2-methoxybenzoic acid,
Limits:
0.10 per cent;
Maximum 0.5 per cent, determined on 1.000 g by drying in F. 4-amino-N-[[(2RSJ-I-ethyl-I-oxidopyrrolidin-2-yl]
an oven at 105°C for 3 h. methyl]-5-(ethylsulfonyl)-2-methoxybenzamide,
ASSAY
Dissolve 0.300 g with shaking in a mixture of 5 mL of acetic
anhydride Rand 50 mL of anhydrous acetic acidR. Titrate
with 0.1 M perchwnc acid, detennining the end-point
G. 4-amino-N- [(3RSJ -J-ethylpiperidin-3-yl]- 5-( ethylsulfonyl)-
potentiometrically (2.2.2(J).
z-merhosvbeneamlde,
I mL of 0.1 M perchloril; acid is equivalent to 36.95 mg of
CI7H21N30"S.
IMPURITIES
and eoanllomer
Speafied impurities A.
Otherdeteerable impurities (thefollowing substances would, if
present at a sufficremlevel, be detected by oneor other 0/the tests
in the monograph. They are limited by thegeneral acceptance H. 4-amino-N-[[(2RSJ-I-ethylpyrrolid in-2-yl]methyl]-5-
cruerion for omerlumpecified impurities and/or by the general (ethylsulfonyl)-2-methoxy-N-methylbenzamide.
monograph Subsranees for pharmaceutical use (2034). It is __ ~ Phf<l
therefore not n«essary to identify these impuniies for
in substances for pharmaceutical use) B, C, DJ E, F, G, H.
Amitriptyline Hydrochloride
(Ph. Bur. monograph 0464)
A. [(2RSJ-I-ethylpyrrotidin-2-yl]methanamine,
313.9 549-18-8
Action and use

B. 4-amino-N-[[(2RSJ-l-ethylpyrrolidin-2-yl]methyl]-5-
Monoa~e reuptake inhibitor; tricyclic antidepressant.
(ethylsulfonyl)-2-hydroxybenzamide,
Preparations
D Amitriptyline Tablets
I HN~
Amitriptyline Oral Solution
'QoP"" . N and enanllomer
H,N "'- DC", H ~ PhE" _
CH,
DEFINITION
3-( I0,II-DihYdro-5H-dibenzo[a,d] [7]annulen-5-ylidene)-N,
C. 4-amino-N-[[(2RSJ -J-ethylpyrrolidln-2-yl]methyl]-5-iodo-
N-dimethylpropan-I-amine hydrochloride.
2-methoxybenzamide,
Content
and enanUomer
CHARACTERS
Appearance
White or almostwhite powder or colourless crystals.
Solubility
D.4-amino-N-[[(2RSJ-I-ethylpyrrolidin-2-yl]methyl]-2- Freely soluble in water, in ethanol (96 per cent) and in
methoxy-5-(methylsulfonyl)benzamide, methylene chloride.
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1-158 Amitriptyline Hydrochloride 2022
IDENTIFICATION - disregard limit: 0.5 times the area of the peakdue to

A. Infrared absorption spectrophotometry (2.2.24). amitriptyline in the chromatogram obtained with reference
Comparison amimPty/ine hydroch/orid< CRS. solution (b) (0.05 per cent).
B. 20 mg gives reaction (a) of chlorides (2.3.1). Loss on drying (2.2.32)
TESTS an oven at 105°C for 2 h.
The solution is clear (2.2.1) and not more intenselycoloured
than reference solutionB1 (2.2.2, Method If). Maximum 0.1 per cent, determined on 1.0 g.
Dissolve 1.25 g in water R and dilute to 25 mL with the ASSAY
same solvent. Dissolve 0.250 g in 30 mL of ethanol (96 per cenv R. Titrate
with 0.1 M sodium hydroxide, determining the end-point
potentiomettically (2.2.20).
Dissolve 0.20 g in carlHm dioxide-jree wener R and dilute to
10 mL with the same solvent. Add 0.1 mL of methyl red I mL of 0.1 M sodium hydroxide is equivalent to 31.39 mg of
solution Rand 0.2 mL of 0.01 M sodium hydroxide. C,oH,.CIN.
The solution is yellow. Add 0.4 mL of 0.01 M hydrochlon'c STORAGE
add. The solution is red. Protected from light.
Related substances IMPIJRITIES
Specified impurities A, B.
Test sdution Dissolve 50.0 mg of the substance to be Otherdetectable impurities (thefollowing substances wouldJ if
examined in the mobilephase and dilute to 50.0 mL with
present at a sufficient levelJ be deuaed by oneor other of the tests
the mobile phase.
in the monograph. They a~ limited by thegeneral acceptance
Reference solution (a) Dissolve 5.0 mg of criterion for otherlunspedfied impurities and/or by thegeneral
dibenzosuberom CRS (impurity A) and 5.0 mg of monograph Substances for pharmaceuucal use (2034). It is
cydDbenzaprine hydrochloride CRS (impurity B) in 5.0 mL of therefore not necessary to identify these impurities for
the test solution and dilute to 100.0 mL with the mobile demonstration of compliance. See also5.10. Control of impurities
phase. in substances for pharmaceutical use) C, D, E, F, G.
Dilute 1.0 mL of reference solution (a)
Reference solution (b)
Golumn:
- size: 1= 0.15 m, 0 = 4.6 mm;
- stationary phase: end-capped po/aNnlbedded octaderylsi/yl
amorphous organosl1i(Q polymer R (5 I!m);
Mobile phase Mix 35 volwnes of aceumilri/e R and
65 volwnes of a 5.23 gIL solution of dipotassium hydrogen A. 10,ll-dihydro-5H-dibenzo[a,d] [7]annulen-5-one
phosphate R previously adjusted to pH 7.0 with phosphoric (dlbenzosuberone),
acid R.
Detection Spectrophotometer at 220 run. -, >""- »<: N ... CH3
Injection I0 ~L. I
CH,
Run time 3 times the retention time of amitriptyline.
Relativeretention With reference to amitriptyline (retention
time = about 14 min): impurity B = about 0.9;
=
impurity A about 2.2. B. 3-(5H-dibenzo[a,d] [7]annulen-5-ylidene)-N,N-
System suitability Reference solution (a): dimetllylpropan-l-amine (cyclobenzaprine),
impurity B and amitriptyline.
Limits:
- impurity B: not more than the area of the corresponding
solution (b) (0.1 per cent);
- impurity A: not more than 0.5 times the areaof the
reference solution (b) (0.05 per cent); C. 3-(10, II-dihydro-5H-dibenzo[a,d] [7]annulen-5-ylidene)-
- unspecified impuritks: for each impurity, not more than the N-methylpropan-I-amine (nortriptyline),
area of the peakdue to amitriptyline in the chromatogram
obtained with reference solution (b) (0.10 per cent);
- toud: not more than 3 times the area of the peak due to
amitriptyline in the chromatogram obtained with reference
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2022 Amlodipine Besilate 1-159
PhEm _ _ ~ _
"'\ -C"'
?6
---" 0" DEFINITION
3-Ethyl 5-methyl (4RS)-2-[(2-aminoethoxy)methyl]-4-(2-
chiorophenyl) -6-me thyl-I ,4-<1ihydropyridine-3,5-dicarboxylate
,:7 j benzenesulfonate.
~ .
Content
D. 5-[3-(dimethylamino)propyl]-10,1 I -dihydro-5H-dibenzo
[a,d][7]annulen-5-01, PRODUCTION
It is considered that alkyl benzenesulfonate esters are
genotoxicand are potential impurities in amlodlpine besilate.
The manufacturing process should be developed taking into
consideration the principles of quality risk management,
together with considerations of the quality of starting
materials, process capability and validation. The general
method 2.5.41. Methyl, ethyland isopropyl bensenesulfonou in
activesubstances is available to assist manufacturers.
E. N,N-dimethyl-3-(1,2,3,4,4a,10, I 1,1Ja-octahydro-5H- CHARACTERS
dibenzo [a,d][7Jannulen-5-ylidene)propan-I -amine, Appearance
White or almost while powder.
Solublllty
Slightly soluble in water, freely soluble in methanol, sparingly
its (E}-isomarand soluble in anhydrous ethanol, slightly soluble in 2-propanol.
theirenanliomers
IDENTIFICATION
Comparison amlodipine besikue CRS.
F. (5EZ, IORS)-5-[3-(dimethylamino)propylidene)-10,11-
TESTS
dihydro-5H-dibenzo[a,d] [7)annulen- I 0-01,
Optical rotation (2.2.7)
-0.10' to + 0.10'.
Dissolve 0.250 g in methanol R and dilute to 25.0 mL with
the same solvent.
Related substances
Liquid chromatography (2.2.29). Cany ou' the zes prouaed
from ligh<
Tes, solution (a) Dissolve 50.0 mg of the subsrance to be
G. 10, I I -dihydro-5H-dibenzo[a,d] [7]annulen-5-01 examined in the mobile phase and dilute [0 50.0 mL with
(dibenzosuberol). the mobile phase.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEm Tes, solution (b) Dilute 5.0 mL of test solution (a) to
100.0 mL with the mobile phase.
Refermce souaion (a) Dilute 1.0 mL of test solution (a) to
Amlodipine Besilate solution to 100.0 mL with the mobile phase.
Reference solmion (b) Dissolve 2.5 mg of amlodipine
(ph. Eur. monograph 1491) impurity B CRS and 2.5 mg of amlodipine impurity G CRS in
the mobile phase and dilute to 25.0 mL with the mobile
phase. Dilute 1.0 mL of the solution to 10.0 mL with the
mobilephase.
Reference tdution (c) Dissolve 2.5 mg of amlodipine for peak
identification CRS (containing impurities D, E and F) in
5 mL of the mobile phase.
Reference sohuion (d) Dissolve 5.0 mg of amlodipine
impurity A CRS in acetonilri/e R and dilute to 5.0 mL with
the same solvent. Dilute 1.0 mL of the solution to 100.0 mL
C"H"CIN,O,S 567.1 11147D-99-6 with the mobile phase. Dilute 1.0 mL of this solution to
Action and use
Reference solution (e) Dissolve 50.0 mg of amlodipine
Calcium channel blocker.
besikue CRS in the mobile pbase and dilute to 50.0 mL with
Preparations the mobile phase. Dilute 5.0 mL of the solution to 100.0 mL
AmIodipine Besilate Tablets with the mobile phase.
AmIodipine Oral Solution Column:
- size: I = 0.25 m, 0 = 4.0 nun;
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1-160 Amlodipine Besilate 2022
- 'latUma/)' phase: Of:tadeq/silyl silica gelfor chromatography R criterion for otherhmspecified impurities and/or by thegeneral
(5 um); monograph Substances for pharmaceuucol use (2034)" It is
- temperature: 30 "C. therefore nor necessary to idennfy these impuniies for
lvlobile phase 2.3 gIL solution of ammonium aceUlU R, demonstration of compliance" See also 5.10. Control of impurities
methanol R (30:70 VIV). in substances for phannauutical use) B, G, H.
Flew raU 1.5 mUmin.
Injection 20 ~L of test solution (a) and reference
solutions (a), (b), (c) and (d).
Run time Twice me retention time of amlodipine.
with amlodipine for peak identification CRS and the
the peaks due to impurities D, E and F; use the
identify the peak due to impurity A. A. 3-ethyl 5-methyl (4RS)-4-(2-cWorophenyl)-2-[[2-(l,3-
dioxo-l ,3-dihydro-2H-isoindol-2-yl) ethoxy]methyl)-6-
Relative retention With reference to amlodipine (retention
methyl-I, 4-dihydtopytidine-3,5-<1icarboxylate,
= =
time about 20 min): impurity G about 0.21;
=
impurity B about 0.25; impurity D about 0.5; =
impurity F :::: about 0.8; impurity E = about 1.3.
Systemsuitability Reference solution (b):
impurities G and B.
Limits:
- cotreaion factors: for me calculation of content, multiply
corresponding correction factor: impurity D = 1.7;
impurity F = 0.7;
- impun"ty D: not more than 3 times the area of the
B. 3-ethyI5-methyl (4RS)-4-(2-cWorophenyl)-6-methyl-2-
[[2-[[2-(methylcarbamoyl)benzoyl]amino]ethoxy]methyl]-
- impurily A: not more than 1.5 times the area of the
1,4-dihydropytidine-3,5-dicarboxylate,
corresponding peak in the chromatogram obtained. with
reference solution (d) (0.15 per cent);
- impurities E, F: for each impurity, not more than 1.5 times
the area of the principal peak in the chromatogram
obtainedwith reference solution (a) (0.15 per cent);
- unspecified impun"ties: for each impurity, not more than me
- disregard limir. 0.5 times the area of the principal peak in D. 3-ethyl 5-methyl 2-[(2-aminoethoxy)methyl]-4-(2-
the chromatogram obtainedwith reference solution (a) cWorophenyl)-6-methylpyridine-3,5-<1icatboxylate,
(0.05 per cent); disregard any peak due to benzene
sulfonate (relative retention = about 0.14).
Water (2.5.1Z)
ASSAY
related substances with the followingmodification.
E. diethyl (4R5)-2-[(2-aminoethoxy)methyl]-4-(2-
Inieaion Test solution (b) and reference solution (e). cWorophenyl)-6-methyl-I,4-dihydtopytidine-3,5-
Calculate the percentage content of C2JI31CIN20SS taking dicarboxylate,
into account the assigned content of amlodipine besikue CRS.
STORAGE
IMPURITIES
Specified impuniies A, D, E, F.
Other detectable impurities (the jol/Qwing subslances would, if
in the monograph. They are limited by the general acceplance
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2022 Ammonia 1-161
H
2 changes from red to yellow. Add 1 mL of sodium cobaltinitrue
H:;xN& C O............... NH
solution R. A yellow precipitate is formed.
H,CO I lOCH,
TESTS
: H
o : 0 Solution S
()
~ CI Evaporate 220 mL almost to dryness on a water-bam. Cool,
I andenantiomer add 1 mL of dilute acetic add R and dilute to 20 mL with
"" distilled waler R.
F. dimethyl (4RSJ-2-[(2-aminoethoxy)methyl]-4-(2-
cWorophenyl)-6-methyl-I,4-dihydropyridine-3,5-
Method II).
dlcarboxylare,
To 2 mL add 8 mL of water R.
H,C
Oxldisable substances
Cautiously add, whilstcooling, 8.8 mL to 100 mL of dilute
sulfuric acidR. Add 0.75 mL of 0.002 M porassium
permanganate. Allow to standfor 5 min. The solution
o remains faindy pink.
Pyridine and related substances
Maximum 2 ppm, calculated as pyridine.
.Measure the absorbance (2.2.25) at 252 urn using water R as
G. dimethyI4-(2-cWorophenyl)-2,6-dimethyl-l,4- the compensation liquid. The absorbance is not greater
dihydropyridine-3,5-dicarboxylate, than 0.06.
Carbonates
H H:,clli Maximum 60 ppm.
To 10 mL in a test-tube with a ground-glass neck add
H3CyNy"o~N~
H'CO~O,-/CH,
10 mL of calcium hydroxide solurion R. Stopperimmediately
0 and mix. Any opalescence in the solution is not more intense
o : 0 than thatin a standard prepared at the same time and in the
~CI same manner using 10 mL of a 0.1 gIL solution of anhydrous
V and eoanllomer sodium carbonate R.
Chlorides (2.4.4)
Maximum I ppm.
H.2-[[2-[[(4RSJ-4-(2-cWorophenyl)-3-(ethoxycarbonyl)-5-
Dilute 5 mL of solution S to IS mL with water R.
(methoxycarbonyl)-6-methyl-I,4-dihydropyridin-2-yl]
methoxy]ethyl)carbamoyl)benzoic acid. Sulfates (2.4.13)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf" Maximum 5 ppm.
Dilute 3 mL of solution S to IS mL with dis.Ued water R.
Iron (2.4.9)
Maximum 0.25 ppm.
Strong Ammonia Solution Dilute 4 mL of solution S to 10 mL with water R.
(Ammonia Solution, Concentrated, Ph. Bur. Maximum 20 mgIL.
monograph 0877)
Evaporate 50 mL to dryness on a water-bath and dry at
NH, 17.03 100-105 "C for I h. The residue weighs a maximum of
Preparadon 1 mg.
Dilute Anunonia Solution ASSAY
Phf" _ Weigh accurately a flask with a ground-glass neck containing
50.0 mL of 1 M hydrochlori< acid. Add 2 mL of the substance
DEFINITION
to be examined and re-weigh. Add 0.1 mL of methylred
Content
solution R as indicator. Titrate with 1 M sodium hydroxide
25.0 per cenr mlm to 30.0 per cent mlm.
until the colour changes from red to yellow.
CHARACTERS I mL of 1 M hydrochWric acid is equivalent to 17.03 mg of
Appearance NH,.
Clear,colourless liquid, very caustic.
STORAGE
Solubility
Protected from air, at a temperature not exceeding 20 "C.
Miscible with water and with ethanol (96 per cent).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phfur
IDENTIFICATION
A. Relative density (2.2.5): 0.892 to 0.910.
B. It is strongly alkaline (2.2.4).
C. To 0.5 mL add 5 mL of water R. Bubble air through the
solution and lead the gaseous mixture obtained over the
surface of a solution containing 1 mL of 0.1 M hydrochloric
acid and 0.05 mL of methyl redsolution R. The colour
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1-162 Ammonio Methacrylate Copolymer 2022
Ammonio Methacrylate Copolymer ***** Monomers

** ** Liquid chromatography (2.2.29).
(Type A) *** Solution A Dissolve 3.5 g of sodium perchlorate R in water R
(Ph. Eur. monograph 2081) and dilute to 100 mL with the same solvent.
solvent. To 10.0 mL of this solution add 5.0 mL of
solution A, dropwise, while continuously stirring. Remove the
precipitated polymer by centrifugation. Use the clear
supernatant solution.
Reference solution Dissolve 50.0 mg of ethyl acryIou Rand
10.0 mg of methylmethacrylate R in methanol R and dilute to
50.0 mL with the same solvent. Dilute 1.0 mL of the
solution to 100.0 mL with methanol R. Add 10 mL of this
solution to 5 mL of solution A.
Action and use Column:
Excipient. ~ size: 1= 0.12 m, 0 =4.6 mm;
Phf" _ - stationary phase: irregular end-copped o<tadecy/si/y/ silica gel
for chromawgraphy R (7 urn).
DEFINITION
Mobile phase Mix 20 volumes of methanol R2 and
Poly[ethyl propenoate-co-methyl z-rnethylprop-z-enoete-ec-N,
80 volumes of water for chromatography R previously ajusted
N,N-trimethyl-2-[(2-methylprop-2-enoyl)oxy]ethan-l-
to pH 2.0 with phosphoric acidR.
aminiwn chloride] having a mean relative molecular mass of
about 150 000. Flow rau 2.0 mIJmin.
The ratio of ethyl acrylate (ethyl propenoate) groups to Detection Spectrophotometer at 202 om.
methyl methacrylate (methyl 2-methylprop-2-enoate) groups Injecrion 50~.
to ammonia methacrylate (N,N,N-trimethyl-2-[(2- System suilability Reference solution:
methylprop-2-enoyl)oxy]ethan-I-aminium chloride) groups is - resolution: minimum 15 between the peaks due to
about 1:2:0.2. impurity A and impurity B.
Content of ammonia memauylale groups 8.9 per cent to Limits:
12.3 per cent (dried substance). - rmpun~ A: not more than the area of the corresponding
CHARACTERS peak in the chromatogram obtained with the reference
solution (100 ppm);
Appearance
Colourless to whiteor almost whitegranules or powder.
- impurily B: not more than 2.5 times the area of the
Solubility the reference solution (50 ppm).
Practically insoluble in water, freely soluble in anhydrous
Methanol (2.4.24, System A)
ethanol and in methylene chloride giving clear to cloudy
solutions. Due to the polymeric nature of the substance) a
stirring time of up to 5 h may be necessary. Loss on drying (2.2.32)
IDENTIFICATION vacuo at 80°C for 5 h.
Comparison ammonio methacrylate copolymer CRS. ASSAY
Dissolve 1.000 g in 75 mL of glacial acetic acid R at about
B. Viscosity (see Tests). 50°C wil:hin about 30 min. Allow to cool to room
C. It complies with the limits of the assay. temperature and add 25 mL of a 6 gIL solution of copper
TESTS acetate R in glacial acelu acidR. Titrate with O. J M perchlonc
Solution S add, determining the end-point potentiometrically (2.2.20).
Dissolve a quantity of the substance to he examined I mL of 0.1 M'perchloric acid is equivalent to 20.77 mg
corresponding to 12.5 g of the driedsubstance in a mixture of c.H IS 0 2 N C I (ammonio methacrylate groups).
of 35.0 g of awone Rand 52.5 g of 2-propanol R. IMPURITIES
Viscosity (2.2.11J) Specified impuri.ies A, B.
Maximum 15 mf'a-s, determined on solution S.
Apparatus Rotating viscometer.
Dimensions:
= =
- spindle: diameter 25.15 mID; height 90.74 mID; shaft
diameter = 4.0 nun; A. ethyl propenoate (ethyl acrylate),
- cylinder. diameter = 27.62 mID; height = 0.135 m.
Srirring speed 30 r/min.
Volume of solution 16 mL of solution S.
Temperature 20 'C.
Appearance of a film
Spread 2 mL of solution S evenly on a glass plate. Upon B. methyl 2-methylprop-2-enoate (methyl methacrylate).
drying a clear film is formed. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf"
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2022 Ammonio Methacrylate Copolymer 1-163
Ammonio Methacrylate Copolymer ***** Monomers

** ** Liquid chromatography (2.2.29).
(Type B) *** Solution A Dissolve 3.5 g of sodium perchlorate R in water R
(Ph. Eur. monograph 2082) and dilute to 100 mL with the same solvent.
Testsolution Dissolve 5.00 g of the substance to be
solvent. To 10.0 mL of this solutionadd 5.0 mL of
solution A, dropwise, while continuously stirring. Remove the
precipitated polymer by centrifugation. Use the clear
supernatant solution.
Reference solu.wn Dissolve 50.0 mg of elhyl aayJateRand
10.0 mg of methylmethacrylate R in methanol R and dilute to
solution to 100.0 mL with mahand R. Add 10 mL of this
solution to 5 mL of solution A.
Excipient. - size: 1= 0.12 m, 0 = 4.6 rnm;
PhE" _ - suuionory phase: irregular end-capped o<tade<y/silyl silica gel
for ehromatography R (7 urn).
DEFINITION Mobile phase Mix 20 volumes of methanol R2 and
Poly[ethyl prcpenoate-ce-methyl 2-methylprop-2-enoate-eo-N, 80 volumes of waterfor chromatography R previously ajusted
N,N-trimethyl-2-[(2-methylprop-2-enoyl)oxy)ethan-l-
aminium chloride] havinga mean relative molecular mass of
'0 pH 2.0 with phosphorie add R.
Flow rare 2.0 mIlmin.
about 150000.
The ratio.of ethyl acrylate (ethyl propenoate) groups to
methyl methacrylate (methyl 2-methylprop-2-enoate) groups Injection 50 ~L.
to ammonio methacrylate (N,N,N-trimethyl-2-[(2- System suitability Reference solution:
methylprop-2-enoyl)oxy)ethan-I-aminium chloride) groups is - resolution: minimum 1.5 between the peaks due to
about 1:2:0.1. impurity A and impurity B.
Content 0/ammonio methacrylate groups 4.5 per cent to Limits:
7.0 per cent (dried subsiance). - impurity A: not more than the area of the corresponding
peak in the chromatogram obtained with the reference
CHARACTERS
solution (100 ppm);
Appearance
- impurity B: not more than 2.5 times the area of the
Colourless to white or almost white granules or powder.
Solubility the reference solution (50 ppm).
Methanol (2.4.24, System A)
ethanol and in methylene chloride giving clearto cloudy
solutions. Due to the polymeric nature of the substance, a
stirring time of up to 5 h may be necessary. Loss on drying (2.2.32)
IDENTIFICATION vacuo at 80°C for 5 h.
A. Infrared absorption spectrophotometry (2.2.24}.
ASSAY
Comparison ammonia methacrylate copdymerCRS.
Dissolve 2.000 g in 75 mL of glaeial aced: add R at about
B. Viscosity (see Tests). 50°C withinabout 30 min. Allow to cool to room
C. It complies with the limits of the assay. temperature and add 25 mL of a 6 gIL solutionof copper
TESTS acetate R in glacial acetic acid R. Titrate with 0.1 M percbtoric
Solution S acid, determining the end-point potentiometrically (2.2.20).
Dissolve a quantity of the substance 10 be examined I mL of 0.1 M perehlorie add is equivalent to 20.77 mg
corresponding to 12.5 g of the dried substance in a mixture of C,Hj,02NCI (ammonio methacrylate groups).
of 35.0 g of aalOlle Rand 52.5 g of 2-propanol R. LID>URITIES
Viscosity (2.2.111) Spedfied impurities A, B.
Maximum 15 mPa·s, determined on solution S.
Apparatus Rotating viscometer.
Dimensions:
-spindle: diameter = 25.15 rnm; height = 90.74 mm; shaft
diameter = 4.0 mm; A. ethyl propenoate (ethyl acrylate),
= =
- cylinder: diameter 27.62 mm; height 0.135 m.
Stirring speed 30 r/min.
o
Volume ofsolution 16 mL of solution S.
H,C,
I'Jl~OCH3
Temperature 20 "C. CH,
Appearance of a film
Spread 2 mL of solution S evenly on a glass plate. Upon B. methyl 2-methylprop-2-enoate (methyl methacrylate).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
drying a clear film is formed.
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1-164 Ammonium Bicarbonate 2022
Ammonium Bicarbonate Ammonium Bromide

(Ammonium Hydrogen Carbonate, Ph. Bur. (Ph. Bur. monograph 1389)
monograph 1390) NH..Br 97.9 12124-97-9
NH..HCO, 79.1 1066-33-7 PhE" _
Action and use DEFINmON

Expectoranr. Content
Preparations
Aromatic Ammonia Solution CHARACTERS
Strong Ammonium Acetate Solution Appearance
Aromatic Ammonia Spirit White or almost white, crystalline powderor colourless
crystals, hygroscopic.
PhE" _
Solublllty
DEFINITION Freelysoluble in water, sparingly soluble in ethanol
Content (96 per cent).
98.0 per cent to 101.0 per cent. It becomes yellow when exposed to light or all'.
CHARACTERS IDENTIFICATION
Appearance A. It gives reaction (a) of bromides (2.3.1).
Fine, white or ahnost white, crystalline powderor white or B. Iu'ml, of solution S (see Tests) gives the reaction of
almost white crystals, slightly hygroscopic. ammonium salts (2.3.1).
Solublllty TESTS
Freely soluble in water, practically insoluble in ethanol Solutlon S
(96 per cent). Dissolve 10.0 g in carbon dioxide-free water R and dilute to
It volatilises rapidly at 60°C. The volatilisation takes place 100 mL with me same solvent.
slowly at ambient temperatures if the substance is slightly
moist. It is in a state of equilibrium with ammonium
Solution S is clear (2.2.1) and colourless (2.2.2, Merhod 11).
carbamate.
IDENI1FICATION To 10 mL of solution S add 0.05 mL of merhyl red solution R.
A. It gives the reaction of carbonates and bicarbonates Not more than 0.5 mL of 0.01 M hydrochloric acid or 0.01 M
(2.3.1). sodium hydroxide is required to change the colour of the
B. Dissolve 50 mg in 2 mL of waterR. The solution gives the indicator.
reaction of ammonium salts (2.3.1). Bromates
TESTS To 10 mL of solution S add 1 mL of su",,11 solution R,
Solutlon S 0.1 mL of a 100 gIL solution of porassium iodide R and
Dissolve 14.0 g in 100 mL of dis.1led waterR. Boil to remove 0.25 mL of 0.5 M sulfuric acid and aUow to stand protected
the ammonia, allow to cool and dilute to 100.0 mL with from light for 5 min. No blue or violet colourdevelops.
distiUed water R. Chlorides and sulfates
Chlorides (2.4.4) Liquid chromatography (2.2.29).
Maximum 70 ppm. Testsolution (aJ Dissolve 00400 g of the substance to be
Dilute 5 mL of solution S to 15 mL with waterR. examined in 50 mL of waterfor chromatography R and dilute
Sulfates (2.4.13)
Maximum 70 ppm, determined on solution S. Tesr solution (b) Dilute 25.0 mL of test solution (a) to
50.0 mL with waterfor chromarography R.
Iron (2.4. 'l)
Maximum 40 ppm. Reference solurion (a) To 25.0 mL of test solution (a) add
1.0 mL of sulfare standard solution (10 ppm SO'> R and
Dilute 1.8 mL of solution S to 10 mL with warer R.
12.0 mL of chloride standard solurion (50 ppm CI) R and dilute
ASSAY to 50.0 mL with waterfor chromawgraphy R.
Dissolve cautiously 0.500 g in 50 mL of carbon dioxide-free Reference solulion (b) Dilute 10 mL oftest solution (a) to
water R. Titrate with 1 M hydrochloric arid, determining the 100 mLwith waterfor chromatographY R. To 2 mL of this
end-point potentiometrically (2.2.20). Read the volume solution add 8 mL of chloride srandard solution (50 ppm CI) R
added at the 2nd point of inflection, or at the point of and dilute to 20 mL with waterfor chromatography R.
inflection if only 1 point is detected.
Blank solurion waterfor chromawgraphy R.
I mL of 1 M hydrochloric acid is equivalent [0 79.1 mg
Column:
of NH.,HCO,.
- size: 1= 0.25 m, 0 = 2 mm;
STORAGE - stationary phase: strongly basic anion-exchange resin for
In an airtight container. chromarography R2 (13 pm),
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r Mobile phase Dissolve 0.600 g of potassium hydroxide R in
waterfor chromawgraphy R and dilute to 1000 mL with the
same solvent.
Flow rate 0.4 mIJrnin.
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2022 Ammonium Chloride 1-165
Detection Conductivity detector equipped with a suitable ion

Ammonium Chloride ***
suppressor. *** ***
Injection 50 ~ of test solution (b), reference solutions (a) (Ph. Bur. monograph 0007) ***
and (b) and the blank solution.
NH.Cl 53.49 12125-02-9
Run time 2.5 times me retention time of bromide.
Retention time Chloride = about 5 min; bromide = about Action and use
8 min; sulfate = about 16 min. Used for the acidification of urine and to correct metabolic
System suitability Reference solution (b): alkalosis.
- resolution: minimum 8.0 between the peaks due to Preparation
chloride and bromide. Ammonium Chloride Mixture
Calculation oj percentage contents: Phf" _
- for chlorides, use the concentration of chloride in
reference solution (a); correct the areaof the peakdue to DEFINITION
chloride in me chromatogram obtained with reference Content
solution (a) by subtracting the area of the peak due to 99.0 per cent to 100.5 per cent (dried substance).
chloride in the chromatogram obtained with test
CHARACTERS
solution (b);
Appearance
- for sulfates, use the concentration of sulfate in reference
White or almost white, crystalline powderor colourless
solution (a); correct the areaof the peak due to sulfate in
crystals.
the chromatogram obtained with reference solution (a) by
subtracting the area of the peak due to sulfatein the Solubility
chromatogram obtainedwith test solution (b). Freelysoluble in water.
Limits: IDENTIFICATION
- chlorides: maximum 0.6 per cent; A. It gives reaction (a) of chlorides (2.3.1).
- sulfates: maximum 0.01 per cent. B. 10 mL of solution S (see Tests) gives the reaction of
Iodides ammonium salts (2.3.1).
To 5 mL of solution S add 0.15 mL ofjenic chloride TESTS
solution Rl and 2 mL of methylene chloride R. Shake and allow
Solution S
to separate. The lower layeris colourless (2.2.2) Method /).
Dissolve 10.0 g in carbon dioxide-free water R prepared from
Iron (2.4.9) distilled water R and dilute to 100 mL with the same solvent.
Maximum 20 ppm.
Dilute 5 mL of solution S to 10 mL with water R. Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
Magnesium and alkaline-earth metals (2.4.7) Acidity or alkalinity
Maximum 200 ppm, calculated as Ca. To 10 mL of solution S add 0.05 mL of methyl red solution R.
10.0 g complieswith the test for magnesium and alkaline- Not more than 0.5 mL of 0.01 M hydrochlorU: acid or 0.01 M
earth metals. The volume of 0.01 M sodium edetate used does sodium hydroxide is required to cbange the colour of the
not exceed 5.0 mL. indicator.
Loss on drying (2.2.32) Bromides and iodldes
Maximum 1.0 per cent, determined on 1.000 g by drying in To 10 mL of solution S add 0.1 mL of dame hydrochloric
an oven at 105 DC. acidRand 0.05 mL of chwramine solution R. After I min, add
Sulfated ash (2.4.14) 2 mL of chloroform R and shake vigorously. The chloroform
Maximum 0.1 per cent, determined on 1.0 g. layer remains colourless (2.2.2, Method /).
ASSAY Sulfates (2.4.13)
Maximum 150 ppm.
Dissolve 80.0 mg in water R, add 5 mL of dilute nioic acid R
and dilute to 50 mL with water R. Titratewith 0.1 M silver Dilute 10 mL of solution S to 15 mL with distilled water R.
nitrate, determining the end-point potentiometrically (2.2.20). Calcium (2.4.3)
1 mL of 0.1 M silvernitrate is equivalent to 9.794 mg of Maximum 200 ppm.
NH.,Br. Dilute 5 mL of solution S to 15 mL with distiUed waterR.
Calculate the percentage content ofN}-4Br using the Iron (2.4.9)
following expression: Maximum 20 ppm.
Dilute 5 mL of solution S to 10 mL with waltr R.
a-2.763b
a percentage content of ~Br and NH,.CIobtained in the assay Maximum 1.0 per cent, determined on 1.00 g by drying in
and calculated as NHtBrj an oven at 105 DC for 2 h.
b percentage content of CI obtained in the test for chlorides.
STORAGE
In an airtight container, protected from light. ASSAY
__ ~ PhE" Dissolve 1.000 gin 20 mL of waterR and add a mixture of
5 mL of formaldehyde solution R, previously neutralised to
phenolphthalein solution R, and 20 mL of water R. After
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1-166 Ammonium Glycyrrhizinate 2022
1-2 min, titrate slowly with 1 M sodium hydroxide, using a Test solution Dissolve 0.100 g of the substance to be
further 0.2 mL of me same indicator. examined in the mobile phase and dilute to 100.0 mL with
I mL of 1 M sodium hydroxide is equivalent to 53.49 mg the mobile phase.
of Nll,CI. Reference solution (a) Dilute 1.0 mL of the test solution to
____________ ~ PhE", 20.0 mL with the mobile phase.
Reference solution (b) Dissolve 50 mg of ammonium
glycyTThizate CRS in the mobile phase and dilute to 50.0 mL
with the mobile phase. Dilute 1.0 mL of the solution to
Ammonium Glycyrrhizinate 20.0 mL with the mobile phase.
Column:
(Ammonium Glycyrrhizate, Ph. Bur. monograph - size: I = 0.25 m, 0 = 4.0 mm,
1772) - slationary phase: oelade<ylsi/yl silica gelfor ehromarograp/ry R
(5-10 urn).
klobile phase glacial acetic acid RJ acetonitrile R, water R
(6:380:614 VIVIV).
Injection 10 fll..
Runrime 3 times the retention time of l8P-glycyrrhizic acid.
Relative retention With reference to 18P-glycyrrhizic acid
(retention time = about 8 min): impurity A = about 0.8j
=
l Sc-glycyrrhlaic acid abour 1.2.
- resolution: minimum 2.0 between me peaks due to 18P-
C-l2H65NOl6 840 53956-04-0 glycyrrhizic acid and I Be-glycyrrhizic acid.
PhE", ~ __ Limits:
- 1&t.-glyeyrrhizic acid: not more than twice the sum of the
DEFINITION areas of the peaks in the chromatogram obtained with
Mixture of ammonium 18~- and 181!-glycyrrhizate reference solution (a) (10.0 per cent),
(ammonium salt of (20P)-3P'[[2-0-(I!-D- - impun·cy A: not more than the sum of the areas of the
glucopyranosyluronic acid)-<t-o-glucopyranosylwonic acid] peaks in the chromatogram obtainedwith reference
oxy]~11-oxoolean-12-en-29-oic acid), the J8Jl-isomer being solution (a) (5.0 per cent),
the main component. - altll other impun·ty: for each impurity) not more than
Content 0.4 times the sum of the areas of the peaks In the
98.0 per cent to 102.0 per cent (anhydrous substance). chromatogram obtained with reference solution (a)
(2.0 per cent),
CHARACTERS
- sumof other impurities: not more than 1.4 times the sum of
Appearance
the areas of the peaks in the chromatogram obtained with
White or yellowish-white, hygroscopic powder.
reference solution (a) (7.0 per cent),
Solubllity - disregard limit: 0.04 times the sum of the areas of the
Slightly soluble in water. very slightly soluble in anhydrous peaks in the chromatogram obtained with reference
ethanol, practically insoluble in acetone. It dissolves in dilute solution (a) (0.2 per cent).
solutions of acidsand of alkali hydroxides.
Water (2.5.12)
IDENIIFICATION Maximum 6.0 per cent) determined on 0.250 g.
A. Infrared absorption spectrophotometry (2.2.24). Sulfated ash (2.4.14)
Comparison ammonium glycyrrhizate CRS. Maximum 0.2 per cent) determined on 1.0 g.
B. Dissolve 0.1 gin 20 mL of waterR, add 2 mL of dilute ASSAY
sodium hydroxide solution R and heat cautiously. On heating, Dissolve 0.600 g in 60 mL of anhydrous acetic acid R heating
the solution gives off vapours that may be identified by the at 80°C if necessary. Cool. Titrate with 0.1 M perchloric acid,
alkaline reaction of wet litmus paper (2.3.1). determining the end-point potentiometrically (2.2.20).
TESTS I mL of 0.1 M perchlmie acid is equivalent to 84.0 mg
Appearance of solution of C42H65N016'
The solution is clear (2.2.1) and not more intensely coloured
STORAGE
than reference solution BY7 (2.2.2, Method1).
Dissolve 1.0 g in ethanol (20 pereen' VII-? R and dilute to
+ 49.0 to + 54.0 (anhydrous substance).
Dissolve 0.5 g in ethanol (50 per een' VII-? R and dilute to
50.0 rnL with the same solvent.
Related substances
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2022 Amorolfine Hydrochloride 1-167
IMPURITIES Buffer solution Dissolve 3.5 g of dipotassium hydrogen

phosphate R in 1000 mL of waterfor chromawgraphy Rand
adjust to pH 7.0 with phosphoric acidR.
examined in mobile phase A and dilute to 20.0 mL with
~
co,~
mobile phase A.
OH Reference solution (a) Dissolve 4 mg of amorolfine for system
HO 0 suitability CRS (containing impurities D, E, I and in n
~
C02~ 0 H~3C' mobile phase A and dilute to 5 mL with mobile phase A.
OH HO
100.0 mL with mobile phase A. Dilute 1.0 mL of this
HO solution to 10.0 mL with mobile phaseA.
OH Reference solution (c) Dissolve 4 mg of amorofjine for peak
identification CRS (containing impurity M) in mobile phase A
A. (4P,ZOP)-3P-[[Z-O-(P-D-glucopyranosyluronic acldj-c-n-
and dilute to 5 mL with mobile phase A.
glucopyranosyluronic acid]oxy)-Z3-hydroxy-ll-oxoolean-
IZ-en-Z9-oic acid (Z4-hydroxyglycyrrhizinic acid). Column:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
- size: 1= 0.15 m, Q) = 4.6 mm;
- stationary phase: end-capped amidohexadecyfsilyl silica gelfor
chromawgraphy R (3 pm).
Mobile phase:
----:.. mobik phase A: acetonitrile RJ, buffer solution, methanol R2
Amorolfine Hydrochloride (5:35:60 VIVIV);
- mobile phase B: buffer solution, aatonim'le Rt, methanol R2
(10:30:60 VIVIV);
Time Mobile phose A Mobile phase B

Hcr (min) (per cent V/YJ (per cent VIJI)
0·2 90 to
2 - 25 90 ..... 0 10 ---> 100

354.0 78613-38-4
Action and use Injection 20 ~L.
Antifungal. Identification of impuniies Use the chromatogram supplied
PhE" _
with amorofjine for system suitabih'ty CRS and the
chromatogram obtainedwith reference solution (a) to
DEFINITION identify the peaks due to impurities D, E, I and J; use the
(ZRS,6SR)-Z,6-Dimethyl-4-[(ZRS)-2-methyl-3-[4-(Z- chromatogram supplied with amorolftne for peak
methylbutan-2-yl)phenyl)propyl]morpholine hydrochloride. identification CRS and the chromatogram obtained with
reference solution (c) to identify the peaksdue to impurity M
Content
(peaks I and 2).
Relativeretention With reference to amorolfine (retention
CHARACTERS time = about 15 min): impurity M (peak I) = about 0.56;
Appearance = =
impurity M (peak 2) about 0.60; impurity D about 0.85;
White or almost white) crystalline powder. = =
impurity J about 0.97; impurity I about 1.05; impurity E
Solubility = =
(peak I) about 1.14; impurity E (peak 2) about 1.17.
Slightly soluble in. water, soluble in methanol and in System suila~ility:
methylene chloride. - resolution: minimum 2.0 between the peaks due to
IDENTIFICATION impurity J and amoroJfine in the chromatogram obtained
A. Infrared absorption spectrophotometry (2.2.24). with reference solution (a);
- signal-to-noise ratio: minimum 20 for the principal peak in
Comparison amorolftne hydrochloride CRS.
me chromatogram obtainedwith reference solution (b).
B. To ZO mg add 4.0 mL of waterR, and acidify with dilute
nitric acid R. A precipitate is formed. Centrifuge, and to
- for each impurity, use the concentration of amorolfine
2 mL of the supernatant add 0.4 mL of silver nitrate
hydrochloride in reference solution (b).
sdation RI. A curdled) white precipitate is formed. Centrifuge
and wash the precipitate with 3 quantities, each of 1 mI..., of Limits:
water R. Suspend the precipitate in 2 mL of water R and add - impurity D: maximum 0.2 per cent;
1.5 mL of ammonia R. The precipitate dissolves easily with - impurity E: maximum 0.2 per cent for the sum of the
me possible exception of a few large particles which dissolve areas of the 2 peaks;
slowly. - impuniy I: maximum 0.15 per cent;
~ impun'ty 1W : maximum O. J5 per cent for each peak;
TESTS - unspecified impurities: for each impurity, maximum
Related substances 0.10 per .cent;
Liquid chromatography (2.2.29). - total: maximum 0.4 per cent;
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1-168 Amorolfine Hydrochloride 2022

Sulfated ash (2.4.14) itseplmer al C' andtheirenanliomers
ASSAY E. mixture of (2RS,6RSJ-2,6-dimethyl-4-[(2R)-2-methyl-3-
Dissolve 0.250 g in a mixture of 10.0 mL of 0.01 M [4-(2-methylbulan-2-yl)phenyl]propyl]morpholine and
hydrochloric acid and 40 mL of "hanoi (96 per<enQ R. Carry (2RS,6RS) -2,6-dimethyl-4-[(2SJ-2-methyl- 3- [4-(2-
om a potentiometric titration (2.2.20), using 0.1 M sodium methylbutan-2-yl)phenyl]propyl)morpholine,
inflexion.
1 mL of 0.1 M sodium hydroxide is equivalent 10 35.40 rng of
C2.H"CINO.
IMPURITffiS
Specified impun"ties D, E, I, M.
Otherdetectable impurities (the following substances would, if F. 1-[4-(2-methylbutan-2-yl)phenyl]propan-I-one,
present at a sufficient/evd, be detected by oneor other of the tests
in the monograph. They arelimited by the general aaeptance
criterion for other/unspecified impurities and/or by the general
monograph Substances for pharmaceurical we (2034). It is
therefore not ne«ssary to identify these impurities for
demonstration of compliance. See also 5.10. Control of impun"ties
in substances for pharmaceutical use) A" B, C, P, GJ H, J, s;
L, O.
G. (2RSJ-3-[(2RS,6SR)-2,6-dimethylmorpholin-4-yl]-1-[4-(2-
methylbulan-2-yl)phenyl]-2-methylpropan-l-one,
A. (2RS,43,6SR)-2,6-dimethyl-4-[(2RSJ-2-methyl-3-[4-(2-
methylbutan-2-yl)phenyl)propyl)morpholine 4-oxide,
H. 2-[(2RS)-3-[(2RS,6SR)-2,6-dimethylmorpholin-4-yl]-2-
methylpropyl]-5-(2-methylbutan-2-yl)phenol,
itsepimar at C* and Ihelrenanliomers
B. mixture of (2RSJ-I-[N-[(2R)-2-methyl-3-[4-(2-
methylbulan-2-yl)phenyl]propyl]formamido)plOpan-2-yl
acetate and (2RSJ-I-[N-[(2SJ-2-methyl-3-[4-(2- I. (2RS,6SR)-2,6-dimethyl-4-[(2RSJ-2-methyl-3-[4-[(23)-3-
methylbutan-2-yl)phenyl)plOpyl]formamido]propan-2-yl methylbutan-2-yl]phenyl]propyl)morpholine,
acetate,
H'CQ~
H CH3 andenantiomer
C. (2RS,6SR)-2,6-dimethyl-4-[(2RSJ-2-methyl-3- J. (2RS,6SR)-2,6-dimethyl-4-[(2RSJ-2-methyl-3-[3-(2-
phenylpropyl]morpholine, methylbutan-2-yl)phenyl]propyl)morpholine,
H'C~Q~CH'
H CH3 H3C CH3
and enantlomer
D. (2RS,6SR)-2,6-dimethyl-4-[(2RSJ-3-(4-lerr-burylphenyl)- K. (2RS,6SR)-2,6-dimethyl-4-[(2RS)-2-methyl-3-[4-(3-
2-methylpropyl)morpholine, methylpentan-3-yl)phenyl]propyl]morpholine,
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2022 Arnoxicillin Sodium 1-169
H,C CH,
H Solubility
,p CH,
Very soluble in water, sparingly soluble in anhydrous ethanol,
H'CQ very slightly soluble in acetone.
H CH,
H CH, H,C
'" IDENTIFICATION
H,C First identification: A, D.
and enanbomer CH, Second identification: B, C, D.
L. (2RS,6SR)-4-[(2RS)-3-[3,5-bis(2-methylbutan-2-yl)
phenyl]-2-methylpropyl]-2,6-dimethylmorpholine, Preparation Dissolve 0.250 g in 5 mL of water R, add
0.5 mL of dilute acetic acid R, swirl and allow to stand for
10 min in iced water. Filter the crystals and wash with
2-3 mL of a mixture of 1 volume of water Rand 9 volumes
of acetone R, then dry in an oven at 60 DC for 30 min.
Comparison amoxicillin rrihydrau CRS.
its ep;mer at C' and lhelr enanOOmers Test solution Dissolve 25 mg of the substance to be
examined in 10 mL of sodium hydrogen carbonate solution R.
M.mixture of (IRS,2RS)-3-[(2RS,6SR)-2,6- Reference solution (a) Dissolve 25 mg of amoxicillin
dimethylmorpholin-4-yl]-2-methyl-I-[4-(2-methylbutan-2- lrihydrate CRS in 10 mL of sodium hydrogetl carbonate
yl)phenyljpropan-I-ol and (IRS,2SR)-3-[(2RS,6SR)-2,6- solution R.
dimethylmorpholin-4-yl]-2-methyl-I-[4-(2-methylbutan-2-
Reference solution (b) Dissolve 25 mg of amoxicillin
yl)phenyl]propan-I-ol,
lrihydrote CRS and 25 mg of ampid/lin 'rihydrate CRS in
10 mL of sodium hydrogen carbonate solution R.
Plate TLC silanised silita gel plate R.
Mobl7e phase Mix 10 volumes of acetone Rand 90 volumes
of a 154 gIL solution of ammonium acetaU R previously
adjusted to pH 5.0 with glacial acetic add R.
Applicolifm I ~L.
O. (2RS,6SR)-2,6-dimethyl-4-[(2RS)-2-methyl-3-[4-(propan- Development Over a path of 15 em,
2-yl)phenyl]propyl]niorpholine. Drying In air.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ F'fJE"
Dueaion Expose to iodine vapour until the spots appear
and examine in daylight.
System sutiability Reference solution (b):
- the chromatogram shows 2 clearly separated SpOlS.
Amoxicillin Sodium Results The principal spot in the chromatogram obtained
with the test solution is similar in position, colour and size to
(Ph. Eur. mollograph 0577) the principal spot in the chromatogram obtained with
H reference solution (a).
o XC~Na
HO 0 · I
~
H, NH2
0
~··t-t-s
H H
.x
)---";1-' CH3
CH,
c. Place about 2 mg in a test-tube about 150 nun long and
about 15 nun in diameter. Moisten with 0.05 mL of water R
and add 2 mL of sulfur/< acid-formaldehyde reagent R. Mix the
contents of the tube by swirling; the solution is practicaUy
colourless. Place the test-tube in a water-bath for 1 min;
a dark yellow colour develops.
387.4 14642-77-8
D. It gives reaction (a) of sodium (2.3.1).
Action and use TESTS
Penicillin antibacterial. Appearance of solution
Preparations The solution is not more opalescent than reference
Amoxicillin Injection suspension II (2.2.1), it may show an initial, but transient,
Co-amoxiclav Injection pink colour, and after 5 min, its absorbance (2.2.25) at
430 run is not greater than 0.20.
PhEu _
Dissolve 1.0 g in water R and dilute to 10.0 mL with the
DEFINITION same solvent. Examine immediately after dissolution.
Sodium (2S,5R,6R)-6-[[(2R)-2-amino-2-(4- pH (2.2.3)
hydroxyphenyl) acetyljaminoj-3,3-dimethyl-7-oxo-4-thia-I- 8.0 to 10.0.
azabicyclo [3. 2.Ojheptane-2-carboxylate. Dissolve 2.0 gin carbon dioxide-free waterR and dilute to
Semi-synthetic product derived from a fermentation product. 20 mL with the same solvent.
Content Specific optlcal rotatlon (2.2.7)
89.0 per cent to 102.0 per cent (anhydrous substance). + 240 to + 290 (anhydrous substance).
CHARACTERS Dissolve 62.5 mg in a 4 gIL solution of potassium hydrogen
Appearance phthalate R and dilute to 25.0 mL with the same solution.
White or almost white, very hygroscopic, powder.
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1-170 Amoxicillin Sodium 2022
Related substances Rdative retention With reference to amoxicillin:

Liquid chromatography (2.2.29). = = =
impurity C about 3.4; impurity J (n I) about 4. I;
Test solution (a) Dissolve 30.0 mg of the substance to be impurity J (n = 2) = about 4.5.
examined in mobile phase A and dilute to 50.0 mL with System suitabIlity Reference solution (b):
mobilephase A. - resolution: minimum 2.0 between the peaks due to
Testsolution (b) Dissolve 30.0 mg of the substance to be amoxicillin and cefadroxil; if necessary, adjust the ratio A:
examined in mobile phase A and dilute to 20.0 mL with B of the mobile phase.
mobile phase A.1'repIJre immediately before use. Limits:
Reference solution (aJ Dissolve 30.0 mg of amoxiciUin =
- impurityJ (n I): not more than 3 times the area of the
trihydra/< CRS in mobile phase A and dilute to 50.0 mL with principal peak in the chromatogram obtained with
mobile phase A. reference solution (c) (3 per cent);
- any other impurity: for each impurity, not more than twice
Reference 'olution (b) Dissolve 4.0 mg of e.fadroxilCRS in
mobile phase A and dilute to 50 mL with mobile phase A.
obtained with reference solution (c) (2 per cent);
To 5.0 mL of this solution add 5.0 rnL of reference
solution (a) and dilute to 100 mL with mobile phase A.
in the chromatogram obtained with reference solution (c)
Reference solution (c) Dilute 2.0 mL of reference solution (a) (9 per cent);
to 20.0 mL with mobile phase A. Dilute 5.0 rnL of this - disregard limit: 0.1 times the area of the principal peak in
solution to 20.0 mL with mobile phase A. the chromatogram obtained with reference solution (c)
Reference 'olution (d) To 0.20 g of amoxiciUin trihydrate R (0.1 per cent).
add 1.0 mL of water R. Shake and add dropwise dilute sodium N,N-Dlmethylanillne (2.4.26, Merhad A or B)
hydroxide solution R to obtain a solution. The pH of the
Maximum 20 ppm.
solution is about 8.5. Store the solution at room temperature
for 4 h. Dilute 0.5 rnL of this solution to 50.0 rnL with 2-Ethylhexanoic acid (2.4.21J)
mobile phase A. Maximum 0.8 per cent mlm.
Column: Water (2.5.12)
- size: 1= 0.25 m, 0 = 4.6 mm; Maximum 3.0 per cent, determined on 0.400 g.
- Slationary phase: octade<y/silyl silica gelfor chromawgraphy R Bacterial endotoxin. (2.6. 14)
(5 urn). Less than 0.25 IU/mg)if intended foruse in the manufacture
Mobile phase: of parenteral preparations without a further appropriate
- mobile phase A: mix I volume of acetonitrile Rand procedure for the removal of bacterial endotoxins.
99 volumes of a 25 per cent VIV solution of 0.2 M ASSAY
potassium djhydrogen phospha/< R adjusted to pH 5.0 with Liquid chromatography (2.2.29) as described in the test for
dilute sodium hydroxide solution Rj related substances with the following modifications.
- mobile phase B: mix 20 volumes of aceeonirri!e Rand
Mobile phase Initial composition of the mixture of mobile
80 volumes ofa 25 per cent VIV solution of 0.2 M
phases A and B, adjusted where applicable.
potassium dihydrogen phosphate R adjusted to pH 5.0 with
diJuu sodium hydroxide solution Rj Injection Test solution (a) and reference solution (a).
Systemsuitability Reference solution (a):
Time MobUe phose A MobUe phase B ----: repeatability: maximum relative standard deviation of
(min) (per cent V/V) (per cent VIJI) 1.0 per cent after 6 injections.
0- tR 92 8 Calculate the percentage contentof amoxicillia sodium by
tR· ('R + 25) 92 ...... 0 8 ...... 100 multiplying the percentage content of amoxicillin by 1.060.
(lR + 25) - ('R + 40) 0 (00
(rR + 40) - (IR + 55) 92 8
STORAGE
In an airtight container. If the substance is sterile, store in a
tR = retention time ofamoxicill.in determined with reference solution (e)
sterile, airtight) tamper-evident container.
If the mobilephase has been adjusted to achieve the required IMPURITIES
resolution, the adjusted composition will apply at timezero
in the gradient and in the assay.
Injecuim 50 ~L of reference solutions (b) and (c) with
isocratic elution at the initial mobilephase composition and A. (2S,5R,6R)-6-amino-3,3-<1imethyl-7-oxo-4-thia-l-
50 ~L of test solution (b) and reference solution (d) a211bicyclo[3.2.0]heptane-2-carboxylic acid
according to the elution gradient described under Mobile (6-aminopenicillanic acid),
phase; inject mobilephase A as a blank according to the
elutiongradient described under Mobilephase.
Identification 0/impurities Use the chromatogram obtained
with reference solution (d) to identify the 3 principal peaks
eluted after the mainpeak corresponding to impurity C,
amoxicillin dimer (impurity Jj n = 1) and amoxicillin trimer
(impurity J; n = 2).
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2022 Amoxicillin Trihydrate 1-171
HO m. I
#
H NH 2
a
o
~-#S
H H
N
~H\
COli
CH3
CH,
B. (2S,5R,6R)-6-[[(2S)-2-amino-2-(4-hydroxyphenyl)acetylj H. (2R)-2-[(2,2-dimethylpropanoyl)aminoj-2-(4-
aminoj-3,3-dimethyl-7-oxo-4-thia-l-azabicyclo[3.2.0j hydroxyphenyl) acetic acid,
heptane-2-earboxylic acid (L-amoxicillin),
I. (2R)-2-amino-2-(4-hydroxyphenyl)acetic acid,
C. (4S) -2- [5-(4-hydroxyphenyl)-3,6-dioxopiperazin-2-ylj-5,5-

dimethyllhiazolidine-4-carboxylic acid (amoxicillin
diketopiperazjnes},
I-! C~H
·
H NH2
H.AX
HNX--CH3
HO 0I
~
0
N s CH 3
ICo,H
D. (4S)-2-[[[(2R)-2-amino-2-(4-hydroxyphenyl)acetylJ J. co-oligomers of amoxicillin and penicilloic acids of

aminojcarboxymethylj-5,5-dimethylthiazolidine-4- amoxicillin,
carboxylic acid (penicilloic acids of amoxicillin),
HO m
I
· "":::::
H NH
2
o
~
HNX--CH3
,.-['s
I.
H
~ co,H
X CH 3
,nd_plmor"C'
E. (2RS,4S)-2-[[[(2R)-2-amino-2-(4-hydroxyphenyI)
acetyljarninojmethyl]-5,5-dimethylthiazolidine-4- K. oligomers of penicilloic acids of amoxicillin.
carboxylic acid (penilloic acids of amoxicillin), _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
(X0~OH
N I '" Amoxicillin Trihydrate
"'" OH
F. 3-(4-hydroxyphenyl)pyrazin-2-01,
~
: NH,
HO H
-
HNH
00
H # ~.C02H
CH,
o1'
. N·· S CHJ
H H
HO .# 0 61336-70-7
G. (2S,5R,6R)-6-[[(2R)-2-[[(2R)-2-amino-2-(4- Action and use

hydroxyphenyI)acetyljaminoj2-(4-hydroxyphenyl)acetylj Penicillin antibacterial.
aminol-3,3 -dimethyl-7-oxo-d -thia-Lazabicycl0 [3.2.0j Preparations
heptane2-carboxylic acid (D-(4-hydroxyphenyl) Amoxicillin Capsules
glycylamoxiciliin), Amoxicillin Oral Suspension
Co-amoxiclav Oral Suspension
Co-amoxiclav Tablets
Co-amoxiclav Dispersible Tablets
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1-172 Amoxicillin Trihydrate 2022
PIlE" _ Related substances

liquid chromatography (2.2.29).
DEFINITION
(2S,5R,6R)-6-[[(2R)-2-Amino-2-(4-hydroxyphenyl) Buffersolution pH 5.0 To 250 mL of 0.2 M potassium
acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-I-azabicyclo[3.2.0] dihydrogen phospha.. R add dilu.. sodium hydroxide solution R
heptane-2-earboxylic acid trihydrate. to pH 5.0 and dilute (0 1000.0 mL with water R.
Semi-synthetic product derived from a fermentation product. Test solution (aJ Dissolve 30.0 mg of the substance to be
Content
mobile phase A.
Test sdution (b) Dissolve 30.0 mg of the substance to be
CHARACTERS examined in mobile phase A and dilute to 20.0 mL with
Appearance mobile phase A. Prepare immediarely before use.
White or almost white, crystalline powder. Reference solution (a) Dissolve 30.0 mg of amoxiclllin
Solubility ,';hydra.. CRS in mobile phase A and dilute to 50.0 mL with
Slightly soluble in water, very slightly soluble in ethanol mobile phase A.
(96 per cent), practically insoluble in fatty oils. It dissolves in Reference solution (b) Dissolve 4.0 mg of cefadroxil CRS in
dilute acids and dilute solutionsof alkali hydroxides. mobile phase A and dilute to 50 mL with mobile phaseA.
IDENTIFICATION To 5.0 mL of this solution add 5.0 mL of reference
First idennfication: A. solution (a) and dilute to 100 mL with mobile phase A.
Second identification: B, C. Reference solution (c) Dilute 2.0 mL of reference solution (a)
A. Infrared absorption spectrophotometry (2.2.24). to 20.0 mL with mobile phase A. Dilute 5.0 mL of this
solution to 20.0 mL with mobile phaseA.
Comparison amoxidl/in trihydrace CRS.
Column:
B. Thin-layer chromatography (2.2.27). - size: 1 = 0.25 m, 0 = 4.6 mm;
Test solution Dissolve 25 mg of the substance to be - sta'ionary phase: octadecylsilyl sili<. gelfor chromatography R
examined in 10 mL of sodium hydrogen carbonate solution R. (5 pm).
Reference solution (a) Dissolve 25 mg of amoxicil/in Mobile phase:
,';hydra.. CRS in 10 mL of sodium hydrogen carbona te - mobile phase A: acetonitrile R, buffer solutionpH 5.0
solution R. (1:99 VII');
Reference solution (b) Dissolve 25 mg of amoxicilJin - mobik phase B: acetonitn1e RJ buffer solutionpH 5.0
r';hydra.. CRS and 25 mg of ampicillin trihydrace CRS in (20:80 VII');
PIa.. TLC silanised silka gelplace R. Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent VM
MoMe phase Mix 10 volumes of autone Rand 90 volumes
O-'R 92 8
of a 154 gILsolutionof ammonium acetate R previously
IR - ('R + 25) 92 -> 0 8 -> 100
adjusted to pH 5.0 with glacial au,ic acid R.
('R + 25) . (rR + 40) 0 100
Application I pL. (tR + 40) . (rR + 55) 92 8
Development Over a path of 15 em. rR =retention time of arnoxicillin determined with reference solution (c)
Drying In air.
Detection Expose to iodine vapour until the spots appear If the mobile phase composition has been adjusted to achieve
and examine in daylight. the required resolution, the adjusted composition will apply
System suitability Reference solution (b): at time zero in the gradient and in the assay.
- the chromatogram shows 2 clearly separated spots. Flow race 1.0 mUmin.
Results The principal spot in the chromatogram obtained Detection Spectrophotometer at 254 nm.
with the test solutionis similar in position, colour and size to Injection 50 ~L of reference solutions (b) and (c) with
the principal spot in the chromatogram obtained with isocratic elution at the initial mobile phase composition and
reference solution (a). 50 J.lL of test solution (b) according to the elution gradient
C. Place about 2 mg in a test-tube about 150 mm long and described underMobile phase; injectmobile phaseA as a
about 15 mm in diameter. Moisten with 0.05 mL of water R blankaccording to the elution gradient described under
and add 2 mL of su([uri< acid-fonnaldehyde reagen, R. Mix the Mobile phase.
contentsof the tube by swirling; the solutionis practically System suitability Reference solution (b):
colourless. Place the test-tube in a water-bath for 1 min; - resolution: minimum 2.0 between the peaks due to
a dark yellow colourdevelops. amoxicillin and cefadroxil; if necessary, adjust the ratio A:
TESTS B of the mobile phase.
Solution S Limit:
With the aid of ultrasound or gentle heating, dissolve 0.100 g - any impurity: for each impurity, not more than the area of
in carbon dioxide-free waterR and dilute to 50.0 mL with the the principal peak in the chromatogram obtained with
same solvent. reference solution (c) (l per cent).
pH (2.2.3) N,N-DImethylanlllne (2.4.26, MethodA or B)
3.5 to 5.5 for solution S. Maximum 20 ppm.
Specific optical rotation (2.2. 7) Water (2.5.12)
+ 290 to + 315 (anhydrous substance), determined on 1l.5 per cent to 14.5 per cent, determined on 0.100 g.
solution S.
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2022 Amoxicillin Trihydrate 1-173
Sulfated ash (2.4.!f)

ASSAY
HO m'I""
~
H NH:!
0
~ CO;zH
HNX--CH3
~ ......... -p-s
H
t. '><CHJ and epimeralC'

A1.obiJe phase Initial composition of the mixtureof mobile E. (2RS,4S)-2-[[[(2R)-2-amino-2-(4-hydroxyphenyl)
phases A and B, adjusted where applicable. acetyl]amino]methyl]-5,5-dimethylthiazoUdine-4-
Injection Test solution (a) and reference solution (a). carboxylic acid (penilloic acidsof amoxicillln),
~~
~N
Calculate the percentage content of CU#19N305S taking
uno account the assigned content of amoxicillin HO
JJ 6H
trihydrate CRS.
STORAGE F. 3-(4-hydroxyphenyl)pyrazin-2-o1,
IMPURITIES
H
a XC~H
N )<CH3
H,N'#S CH,
H H
G. (2S,5R,6R)-6-[[(2R)-2-[[(2R)-2-amino-2-(4-
A. (2S,5R,6R)-6-amino-3,3-dimethyl-7-oxo-4-thia-l-
hydroxyphenyi)acetyl]amino]-2-(4-hydroxy-phenyl)acetyl)
azabicyclo[3.2.0]heptane-2-carboxyUc acid
aminol-3,3-dimethyl-7-oxo-4-thia-f-azabicyclo[3.2.0]
(6-aminopeniciUanic acid),
heprane-z-carboxylic acid (o-(4-hydroxyphenyl)
glycylamoxicillin),
CH 3 0
H,Cj---<
~c H NH
~Co,H
B. (2S,5R,6R)-6-[[(2S)-2-amino-2-(4-hydroxyphenyl)acetyl) HO~
amino]-3,3-dimethyl-7-oxo-4-thia-l-azabicyclo[3.2.0]
heptane-2-cacboxylic acid (r-emoxicillin), H. (2R)-2-[(2,2-dimethylpropanoyl)amino]-2-(4-
hydroxypheny1)acetic acid,
H NH2
~CO'H
HO~
I. (2R)-2-amino-2-(4-hydroxyphenyl)acetic acid,
C. (4S)-2-[5- (4-hydroxyphenyl)-3,6-dioxopiperazin-2-yl]-5,5-
dimethylthiazolidine-4-carboxylic acid (amoxicillin
diketopiperazines),
D. (4S)-2-[[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino]
carboxymethyl]-5,5-dimethylthiazolidine-4-<oarboxyUc acid
(penicilloic acids of amoxicilljn} J. co-oligomers of amoxicillin and of penicilloic acids of
arnoxicillin,
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1-174 Amphotericin 2022
CHARACTERS
Appearance
Yellow or orange, hygroscopic powder.
Solubility
Practically insoluble in water, soluble in dimethyl sulfoxide
and in propylene glycol, slightly soluble in
dimethylformamide, very slightly soluble in methanol,
K. oligomers of penicilloic acids of amoxicillin,
It is sensitive (0 light in dilute solutions.
H
IDENTIFICATION
o)--~ -,<CO~~, First identification: B., D.
o~"H-S'><CH, Second identification: A J C.
o I.:~ H H A. Ultraviolet and visible absorption spectrophotometry
H-, NH2 H )---~~CH3
0
HO~ ~
N
"ITs CH,
(2.2.25).
Test solution Dissolve 25 mg in 5 mL of dimelhyl sulfoxide R
and dilute to 50 mL with methanol R. Dilute 2 mL of the
solution to 200 mL with methanol R.
L. (2S,5R,6R)-6-[[(2S,5R,6R)-6-[[(2R)-2-amino-2-(4- Spectral range 300-450 urn.
hydroxyphenyl)acetyl] amino]-3,3-dimethyl-7-oxo-4-thia-l- AbsorptUm maxima At 362 nm, 381 nm and 405 run.
azabicyclo[3.2.0]heptane-2-carbonyl]amino]-3,3-dimethyl- Absorbame ratios:
7-oxo-4-thia-I-azabicyclo[3.2.0]heptane-2-carboxylic acid - Am/Am = 0.57 to 0.61;
(6-APA amoxiciUin amide). =
- A 38 I/A4o , 0.87 to 0.93.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" B. Infrared absorption spectrophotometry (2.2.24).
Comparison amphotericin B CRS.
If the spectra obtained show differences, dry the substance to
be examined and reference substance at 60 QC at a pressure
Amphotericin not exceeding 0.7 kPa for I h and record new spectra.
C. To I rnL of a 0.5 gIL solution in dimethyl sulfoxide R, add
(AmphorenCin B, Ph. Eur. monograph 1292)
5 mL of phospho", add R to form a lower layer, avoiding
mixing the 2 liquids. A blue ring is immediately produced at
the junction of the liquids. Mix, an intense blue colour is
produced. Add 15 mL of water R and mix; the solution
becomes paleyellow.
D. Examine the chromatograms obtained in the lest for
related substances.
with the testsolution at 383 run is similar in retention time
to the principal peak in the chromatogram obtained with
TESTS
Related substances
Liquid chromatography (2.2.29). Protect the soludonsfrom light
924 1397-89-3 and use within 24 h of preparation., except for reference
solution (c) which should be injuted immediately afterits
Action and use preparation.
Antifungal. So/vent mixture 10 gIL solution of ammonium acetate RJ
Preparation N-melhylpyrrolidone R, methanol R (1:1:2 VIV/V).
Amphotericin for Infusion Test solution Dissolve 20.0 mg of the substance to be
PIlE" _ examined in 15 mL of N-methylpyrrolidone R and within 2 h
dilute to 50.0 mL with the solvent mixture. Dilute 5.0 mL of
DEFINITION this solution to 25.0 mL with the solvent mixture.
Mixture of antifungal polyenes produced by the growth of Reference solution (a) Dissolve 20.0 mg of
certain strains of Streptomyces nodosus or obtained by any amphotericin B CRS in 15 mL of N-methylpyrrolidmle Rand
othermeans. It consistsmainly of amphotericin B which is within 2 h dilute to 50.0 mL with the solvent mixture. Dilute
(IR,3S,5R,6R,9R, IIR, 15S, 16R,17R, ISS, 19E,2IE,23E,- 5.0 mL of this solution to 25.0 mL with the solvent mixture.
25E,27E,29E,3IE,33R,35S,36R,37 S)-33-[(3-arnino-3,6-
Reference solution (b) Dilute 1.0 rnL of reference solution (a)
dideoxy-Il-D-mannopyranosyl)oxy]-I ,3,5,6,9,11,17,37-
to 100.0 mL with the solvent mixture.
oetahydroxy-15,16, 18-trimethyl-13-oxo-14,39-dioxa-bicyclo
[33.3.1]nonaniaconta-19,21,23,25,27,29,31-heptaene-36- Reference solution (c) Dissolve 20.0 mg of nystatin CRS in
carboxylic acid. 15 rnL of N-merhylpyrrolidone R and within 2 h dilute to
50.0 mL with the solvent mixrure. Dilute 5.0 mL of the
Content solution to 25.0 mL with reference solution (a). Dilute
Minimum 750 ill/mg (dried substance).
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2022 Amphotericin 1-175
2.0 mL of this solution to 100.0 mL with the solvent - impurityB at 383 nm: not more than 4 times the area of
mixture. the principal peak in the chromatogram obtained with
Reference solution (d) In order to prepare impurities Band reference solution (b) (4.0 per cent);
C, dissolve 10 mg of the substance to be examined in 5 mL - any otherimpUl;ty at 383 nm: for each impurity, not more
of N-methylpyrrolidone R and within 2 h add 35 mL of a chan 2 times the area of the principal peak in the
mixture of 1 volume of methanol Rand 4 volumes of chromatogram obtained with reference solution (b)
anhydrous ethanol R. Add 0.10 mL of dilute hydrochloric (2.0 per cent);
acid R, mix and incubate at 25 "C for 2.5 h. Add 10 mL of - totalat 303 and 383 nm: maximum 15.0 per cent;
10 gIL solution of ammonium acetate R and mix. - disregard limit at 303 nm: 0.05 times the area of the
Reference solution (e) Dissolve 4 mg of amphotericin B for
peak identification CRS (containing impurities A and B) in
5 mL of N-methylpynvlidone R and within 2 h dilute to - disregard limit at 383 nm: 0.1 times the area of the
50 mL with the solvent mixture.
reference solution (b) (0.1 per cent).
Blank solution The solvent mixture.
Lnss on drying (2.2.32)
Column: Maximum 5.0 per cent, determined on 1.000 g by drying in
=
- size: 1;;;:; 0.15 m, 0 4.6 mm;
an oven at 60°C at a pressure not exceeding 0.7 kPa.
- stationary phase: base-deactivated end-capped oaadecylsl1yl
silica gelfor chromatography R (3 urn); Sulfated ash (2.4.14)
- temperature: 20 "C. Maximum 3.0 per cent, determined on 1.0 gj if intended for
use in the manufacture of parenteral preparations: maximum
Mobl1e phase:
0-.5 per cent.
- mobile phaseA: mix 1 volwne of methanolR, 3 volumes of
acetonitrile Rand 6 volumes of a 4.2 gIL solution of citric Bacterial endotoxlns (2.6.14)
acid monohydrate R previously adjusted to pH 4.7 using Less than 1.0 IU/mg, if intended for use in the manufacture
concentrated ammonia R; of parenteral preparations without a further appropriate
- mobile-phase B: mix 12 volumes of methanolR, 20 volumes procedure for the removal of bacterial endotoxins.
of a 4.2 gIL soludon of citric acid monohydrate R previously ASSAY
adjusted to pH 3.9 using concentrated ammonia Rand Protea all solutions from light throughout the assay Dissolve
68 volumes of tuetonitriJe R; 25.0 rng in dimethylsulfoxide R and dilute, with shaking, to
25.0 mL with the same solvent. Under constant stirring of
this stock solution, dilute with dimethyl sulfoxide R to obtain
(min) '(per cent JIll? (per cent VIl?
solutions of appropriate concentrations (the following
0-' 100 0 concentrations have been found suitable: 44.4, 66.7 and
3·23 100 ..... 70 o . . . 30 100 IUlmL). Prepare final solutions by diluting I :20 with
23 - 33 70 ..... 0 30 ..... 100
0.2 M phosphate buffer solution pH 10.5 so that they aU
33 - 40 0 100
contain 5 per cent VIV of dintethyl sulfoxide. Prepare the
reference and the test solutions simultaneously. Carry out the
Flow rate 0.8 mUmin. microbiological assay of antibiotics (2.7.2).
Detection Spectrophotometer: Use amphotericin B for microbiological assay GRS as the
- at 303 run: detection of tetraenes; chemical reference substance.
- at 383 nm: detection of heptaenes. STORAGE
Injection 20 ilL of the test solution and reference Protected from light, at a temperature of 2 °C to 8 °C in an
solutions (b), (c), (d) and (e). airtight container. If the substance is sterile, store in a sterile,
Identification oj impun'ties Use the chromatograms supplied tamper-evident container.
with amphotericin B for peak identification GRS and the LABELLING
chromatograms obtained with reference solution (e) to
The label states, where applicable, that the substance is
identify the peaks due to impurities A and B.
suitable for use in the manufacture of parenteral
Relative retention With reference to amphotericin B preparations.
= =
(retention time about 16 min): impurity B about 0.75;
= =
impurity A about 0.8; nystatin about 0.85. IMPURITIES
Specified impuruies A, B.
System suitability at 383 nm Reference solution (d):
- resolution: minimum 1.5 between the 2 peaks presenting a Otherdetectable impun·tres (the following substances would, if
relative retention of about 0.7. present at a sufficient level, be detected by oneor otherof the tests
Limits: criterion for other/unspecified impurities and/or by the general
- impun'ty A at 303 nm: not more than 2.5 times the area of
monograph Substances for phannaceutical use (2034). It is
the principal peak in the chromatogram obtained with
therefore not netessary to identify these impun·tks for
reference solution (c) (5.0 per cent); if intended for use in
the manufacture of parenteral preparations: not more than
in substances for phannaceutical use) C.
me area of the principal peak in the chromatogram
obtained with reference solution (c) (2.0 per cent);
- Qtry" other impurity at 303 nm: for each impurity, not more
(1.0 per cent);
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1-176 Ampicillin 2022
P/lEIl _
DEFINITION
(2S,5R,6R)-6-[[(2R)-2-Amino-2-phenylacetyl]amino]-3,3-
dimethyl- 7-oxo-4-thia-I-azabicyclo[3.2.0]heptane-2-
carboxylic acid.
Semi-synthetic product derived from a fermentation product.
Content
CHARACTERS
Appearance
A. amphotericin A (28,29-dihydro-amphotericin B), Solubility
Sparingly soluble in water, practically insoluble in acetone, in
H pHCo,H ethanol (96 per cent) and in fatty oils. It dissolves in dilute
solutions of acids and of alkali hydroxides.
~
HO-, HHPH HO'. HHPH" --H
. "k 0 j-,
H 'C>H .. 0 H IDENTIFICATION
o 0 H3C First identification: A.. D.
H
Second identification: BJ C, D.
--CH,
CH, Preparation Discs of potassium bromide R.
Comparison anhydrous ampkiJlin CRS.
B. amphotericin XI (l3-D-methyl-amphotericin B),
Reference solution (a) Dissolve 25 mg of anhydrous
H ,.oH~H ampkiJlin CRS in 10 mL of sodium hydrogen carbonate
solution R.
.......
HO H H OH 'HO H H OH kj-H
" Reference solution (b) Dissolve 25 mg of amoxidllin
~
." 0 " trihydrate CRS and 25 mg of anhydrous ampicillin CRS in
H 'OH (0 H 10 mL of sodium hydrogen carbonate solution R.
o 0
H CH3 Plate TLC silanised SIlica gelplate R.
--CH 3 Mobilephase Mix 10 volumes of aatone Rand 90 volumes
of a 154 gIL solution of ammonium acetate R previously
adjusted to pH 5.0 with glacial acetic add R.
Application I flL.
Drying In air.
C. amphotericin X2 (l3-0-ethyl-amphotericin B). Detection Expose to iodine vapour until the spots appear
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P/lEIl and examine in daylight
Ampicillin *** with the test solution is similar in position, colourand size to
** ** the principal spot in the chromatogram obtained with
(Ph. Eur. monograph 0167) ***** reference solution (a).
C. Place about 2 mg in a test-tube about 150 mm long and
H
o
~.++s
~~
C~H
:: about 15 mm in diameter. Moisten with 0.05 mL of ~aler R
cn
H and add 2 mL of su/furi< acid-fonnaldehy<k reagen' R. Mix the
', Nil, contents of the rube by swirling; the solutionis practically
I H H colourless. Place the test-rube in a water-bath for I min;
"'- 0 a dark yellow colourdevelops.
D. Water (see Tests).
69-53-4
TESTS
Action and use Appearance of solution
Penicillin antibacterial. The solutions are not more opalescent than reference
suspension II (2.2. I).
Preparations
Ampicillin Capsules Dissolve 1.0 g in 10 mL of 1 M hydrochloric add. Separately
dissolve 1.0 g in 10 mL of dilute ammonia R2. Examine
Ampicillin Oral Suspension immediately after dissolution.
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2022 Ampicillin 1-177
pH (2.2.3) Limit:
3.5 to 5.5. - any impurity; for each impurity, not more than the area of
Dissolve 0.1 g in carbon dioxide-free wain R and dilute to the principal peak in the chromatogram obtained with
40 mL with the same solvent. reference solution (c) (1.0 per cent).
Specific optical rotation (2.2.7) N,N-Dimethylaniline (2.4.26, Methad B)
+ 280 to + 305 (anhydrous substance). Maximum 20 ppm.
Dissolve 62.5 mg in water R and dilute to 25.0 mL with the Water (2.5.12)
same solvent. Maximum 2.0 per cent, determined on 0.300 g.
Related substances Sulfated ash (2.4.14)
Liquid chromatography (2.2.29). Maximum 0.5 per cent, determlned on 1.0 g.
Test solution (a) Dissolve 27.0 mg of the substance to be ASSAY
examined in mobile phase A and dilute to 50.0 mL with Liquid chromatography (2.2.29) as described in the test for
mobile phase A. related substances with the following modifications.
Test solution (b) Prepare immediately before use. Dissolve l\1.obile phase Initial composition of the mixture of mobile
27.0 mg of the substance to be examined in mobilephase A phasesA and B, adjusted where applicable.
and dilute to 10.0 mL with mobile phase A. Injection Test solution (a) and reference solution (a).
Reference solution (a) Dissolve 27.0 mg of anhydrous System suitability Reference solution (a):
ampicillin CRS in mobile phase A and dilute to 50.0 mL with - repeatability: maximum relative standard deviation of
mobile phase A. 1.0 per cent after 6 injections.
Reference solution (b) Dissolve 2.0 mg of cefradine CRS in Calculate the percentage content of C1JlI9N JO",S from the
mobile phase A and dilute to 50 mL with mobile phase A. declared content of anhydrous ampicillin CRS.
To 5.0 mL of this solution add 5.0 mL of reference
solution (a). STORAGE
Reference solution (e) Dilute 1.0 mL of reference solution (a) In an airtight container, at a temperature not exceeding
30 'C.
Column: IMPURITffiS
- size: 1= 0.25 m, 0 = 4.6 mm;
H
0'1-~--<C~:,
- stationary phase: oclad«y/silyl silica gelfor chromatography R
(5 pm).
Mobile phase: H:2N--H-S'><CH3
- mobile phaseA: mix 0.5 mL of dilute aceti« acidR, 50 mL H H
of 0.2 M potassium dihydrogen phosphate Rand 50 mL of
acetonitrile R, then dilute to 1000 mL with water R; A. (2S,5R,6R)-6-amino-3,3-dimethyl-7-oxo-4-thia-l-
- mobile phase B: mix 0.5 mL of dilute acetic acidR, 50 mL azabicyclo[3.2.0jheptane-2-carboxylic acid
of 0.2 M potassium dihydrogen phosphate Rand 400 mL of (6-aminopeniciUank acid),
acetonitrile R, men dilute to 1000 mL with water R;
Tim. MobUe phase A Mobile phase B

(min) (per cent VIJI) (per cent VIP)
0- ts: 85 15
'R - (rR + 30) 85 ---0 0 15 ..... lOO
(rR + 30) - (rR + 45) 0 \00
(rR + 45) - ('R + 60) 85 15 B. (2S,5R,6R)-6-[[(2S)-2-amino-2-phenylacetyljaminoj-3,3-
dimethyl-7-oxo-4-thia-I-azabicyclo[3.2.Ojheptane-2-
tR = retention lime of ampiciUin determined with reference solution (e)
carboxylic acid (r-ampicillln),
If the mobile phase composition has been adjusted to achieve
the required resolution) the adjusted composition will apply
at time zero in the gradient and in the assay.
Injection 50 ~L of reference solutions (b) and (c) with
isocratic elution at the initial mobile phasecomposition and
50 ~L of test solution (b) according to the elution gradient
described under Mobile phase; inject mobilephase A as a C. (4S)-2-(3,6-dioxo-5-phenylpiperazin-2-yl)-5.5-
blankaccording to the elution gradient described under dimethylthiazolidine-4-carboxylic acid (diketopiperazines
Mobile phase. of ampicillin),
ampicillin and cefradin, if necessary, adjust the ratio A:B
of the mobile phase.
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1-178 Ampicillin Sodium 2022
D. (4S)-2-[[[(2R)-2-amino-2-phenylacetyl) K. (2R)-2-[(2,2-dimethylpropanoyl)amino)-2-phenylacetic
amino] carboxymethyl]-5, 5-dimethyl-1 ,3-thiazolidine-4- acid,
carboxylic add (peniciUoic acids of ampicillin),
.
H C0 2H
HNH2~O++H}-lD
en
?"
""
I
°
..k
H H
S
E. (2R)-2-[[[(2S,5R,6R)-6-[[(2R)-2-amino-2-
CH,
CH,
phenylacetyl) aminoj- 3,3-dimethyl-7 -oxo-d-thia-f-

L. (2R)-2-amino-2-phenylacetic acid (D-phenylglycine),
azabieyclo [3.2.0]hept-2-yl]carbonyl] aminol-2-phenylacetic

acid (ampicillinyl-o-phenylglycine),
F. (2RS,4S)-2-[[[(2R)-2-amino-2-phenylacetyl) M.co-oligomers of ampicillin and of penicilloic acids of

amino] methyl]-5,5-dimethyl-l,3-thiazolidine-4-earboxylic ampicillin.
acid (penilloic acids of ampicillin), _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PbE"
Ampicillin Sodium
G. (3R,6R)-3,6-diphenylpiperazine-2,5-dione, o .., COzNa
H. NHz ')-~~CH'
cYr
~· · H-- s CH,
I H H
"" °
371.4 69-52-3
H.3-phenylpyrazin-2-ol,
Acdon and use
Penicillin antibacterial.
Preparation
Ampicillin Injection
PhE" _
DEFINITION
I. (2S,5R,6R)-6-[[(2R)-2-[[(2R)-2-amino-2- Sodium (2S,5R,6R)-6-[[(2R)-2-amino-2-phenylacetyl)
phenylacetyl]amino]-2-phenylacetyl]amino)-3,3-dimethylamino]-3,3-dimethyl-7-oxo-t-thia-l-szabicyclo[3.2.0]
7-oxo-t-thia-Lazabicyclo [3.2.0)heptane-2-carboxylic acid heptane-2-carboxylate.
(n-phenylglycylamplcillin), Semi-synthetic product derived from a fermentation product.
Content
CHARACTERS
Appearance
White or almost white powder, hygroscopic.
J. (2S,5R,6R)-6-[(2,2-dimethylpropanoyl)antino)-3,3- Solubility
dimethy1-7-oxo-4-thia-l-azabieyclo[3. 2.0[heptane-2- Freelysoluble in water, sparingly soluble in acetone,
carboxylic acid, practically insoluble in fatty oils and in liquid paraffin.
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2022 Ampicillin Sodium 1-179
IDENTIFICATION Related substances

Firsr identification: A, D. Liquid chromatography (2.2.29).
Second identification: B, C, D. TestsolutWn (aJ Dissolve 31.0 mg of the substance to be
A. Infrared absorption spectrophotometry (2.2.24). examined in mobile phase A and dilute to 50.0 mL with
mobile phase A.
Preparation Dissolve 0.250 g in 5 mL of waw R, add
0.5 mL of dilute acelic acid R, swirl and allow to stand for Test solution (b) Dissolve 31.0 mg of the substance to be
10 min in iced water. Filler the crystals through a small examinedin mobile phase A and dilute to 10.0 mL with
sinrered-glass filter (40) (2.1.2), applying suction, wash with mobile phase A. Prepare immediately before use.
2-3 mL of a mixture of 1 volume of water Rand 9 volumes Reference solutian (a) Dissolve 27.0 mg of anhydrous
of acewne R J then dry in an oven at 60°C for 30 min. ampicillin CRS in mobile phase A and dilute to 50.0 mL with
Comparison ampicillin lrihydrate CRS. mobile phase A.
B. Thin-layer chromatography (2.2.27). Reference solution (b)Dissolve 2.0 mg of cefradine CRS in
TestsolutWn Dissolve 25 mg of the substance to be
To 5.0 mL of this solution add 5.0 mL of reference
solution (a).
Reference solution (a) Dissolve 25 mg of ampU;iIlin
trihydrate CRS in 10 mL of sodium hydrogen carbonate
solution R.
Reference solution (d) To 0.20 g of the substance to be
examined add 1.0 mL of water R. Heat the solution at 60 "C
trihydrate CRS and 25 mg of ampicillin trihydrate CRS in
for I h. Dilute 0.5 mL of this solution to 50.0 mL with
mobile phase A.
Plate TLC silanised silica gelplate R.
Column:
Mobile phase Mix 10 volumes of aarone Rand 90 volumes - size: 1= 0.25 m, (2) = 4.6 mm;
of a 154 gILsolution of ammonium acetate R previously - stationary phase: oetadecylsi/yl silica gel for chromatography R
adjusted to pH 5.0 with glacial acetic acid R. (5 pm).
Applicatian I ~L. Mobile phase:
Development Overa path of 15 em. ~ mobile phaseA: mix 0.5 mL of dilute acetic acidR, 50 mL
Drying In air. of 0.2 M potassium dihydrogen phosphate Rand 50 mL of
Detection Expose (0 iodine vapour until the spots appear acetonitrile R, men dilute to 1000mL with Willer R;
and examinein daylight. . - mobile phase B: mix 0.5 mL of dilute acetic acid R, 50 mL
of 0.2 M potassium dihydrogen phosphate Rand 400 mL of
acetonitrile R, then dilute 10 1000 mL with water Rj
Results The principal spot in the chromatogram obtained Time Mobile phase A Mobile phase B
with the test solution is similar in position, colour and size to (min) (per cent V/J? (per cent VIJI)
the principal spot in the chromatogram obtainedwith 0- 'R 85 15
reference solution (a). 'R- (tR + 30) 85 ..... 0 15 -> 100
C. Place about 2 mg in a test-tube about 150 mm long and (IR + 30) - ('R + 45) o 100
about 15 mm in diameter. Moisten with 0.05 mL of water R (tR + 45) - (IR + 60) 85 15
and add 2 mL of su!fu~ acid-formaldehyde reagent R. Mix the 'R = retentiontime of ampicillin derennined with reference solution (c)
contents of the tube by swirling; the solution is practicaUy
colourless. Place me test-tube in a water-bath for 1 min; If the mobile phase composition has been adjusted to achieve
a dark yellow colourdevelops. the required resolution, the adjusted composition will apply
D. It gives reaction (a) of sodium (2.3.1). at time zero in the gradient and in the assay.
TESTS Flow rate 1.0 mUmin.
Appearance of soludon Deuaion Spectrophotometer at 254 om.
Solutions A and B are not more opalescent than reference Injection 50 ~L of reference solutions (b) and (c) with
suspension II (2.2.1) and the absorbance (2.2.25) of isocratic elution at the initial mobilephase composition and
solutionB at 430 nm is not greater than 0.15. 50 ~L of test solution (b) and reference solution (d)
Place 1.0 g in a conical flaskand add slowly and with according to the elution gradient described underMobile
continuous swirling to mL of 1 M hydrochloric acid phase; injectmobile phase A as a blank according to the
(solution A). Separately dissolve 1.0 gin waw R and dilute elution gradient described under Mobile phase.
to 10.0 mL with the same solvent (solution B). Examine Identification of peaks Use the chromatogram obtained with
inunediately afterdissolution. reference solution (d) to identify the peaksdue to ampicillin
pH (2.2.3) and ampicillin dimer.
8.0 to 10.0. Relative reuntion With reference to ampicillin: ampicillin
Dissolve 2.0 g in carbon dioxide-free water R and dilute to dimer = about 2.8.
20 rnLwith the same solvent. Measure 10 min after System suitability Reference solution (b):
dissolution. - resolution: minimum 3.0 between the peaks due to
Specific optical rotation (2.2.7) ampicillin and cefradin; if necessary adjust the ratio A:B
+ 258 to + 287 (anhydrous substance). of the mobilephase.
Dissolve 62.5 mg in a 4 gIL solution of potassium hydrogen
phrhalate R and dilute to 25.0 mL with the same solvent.
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1-180 Ampicillin Sodium 2022
Limits: STORAGE
- ampicillin dime: not more than 4.5 times the area of the In an airtight container. If the substance is sterile, store in a
principal peak in the chromatogram obtained with sterile, airtight, tamper-evident container.
reference solution ee) (4.5 per cent);
IMPURITIES
- any other;mpun"ty: for each impurity, not more than twice
obtained with reference solution (e) (2 per cent).
N,N-Dlmethylanlline (2.4.26, Melhod lJ)
Maximum 20 ppm.
2-Ethylhexanoic acid (2.4.21f)
Maximum 0.8 per cent m/m. A. (2S,5R.6R)-6-amino- 3,3-dimethyl-7-oxo-4-thia-l-
azabicyclo[3.2.0]heptane-2-carboxylic acid
Methylene chloride
(6-aminopenicillanic add),
Gas chromatography (2.2.21f).
Internal standard solution Dissolve 1.0 mL of ethylene o ~ cOzH
chloride R in water R and dilute to 500.0 mL with the same
H ,NH2 \-~~CH3
ctr
solvent.
~ _ · t-t-s CH,
TestsolutWn (aJ Dissolve 1.0 g of the substance to be I H H
examined in waterR and dilute to 10.0 mL with the same "" 0
solvent.
Test solution (b) Dissolve 1.0 g of the substance to be B. (2S,5R,6R)-6-[[(2S)-2-amino-2-phenylacetyl]amino]-3,3-
examined in water R) add 1.0 mL of the internal standard dimethyt-?-oxo-d-thia-f-azablcyclola.2.0] heptaoe-2-
solution and dilute to 10.0 mL with waterR. carboxylic acid (r-ampicillln),
Reference solution Dissolve 1.0 mL of methylene chloride R in
To 1.0 mL of this solution add 1.0 mL of the internal
standard solution and dilute to 10.0 mL with waterR.
Column:
- material: glass;
- size: 1= 1.5 m, '" = 4 mm;
- stationary phase: diatomaCeous earth for gas
chromatography R impregnated with 10 per cent mlm of C. (4S)-2-(3,6-dioKo-5-phenylpiperazin-2-yl)-5,5-
maaogol 1000 R. dimethylthiazolidine-4-carboxylic acid (diketopiperazines
Comer gas nitrogen for chromatography R. of ampicillin),
Flow rale 40 mUmin.
Temperature:
- column: 60°C;
- injection port: 100 DC;
- deteaor: 150 DC.
Calculate the content of methylene chloride taking its density D. (4S)-2-[[[(2R)-2-amino-2-phenylacetyl]
at 20 °C to be 1.325 g1mL. amino]carboxymethyl]-5,5-dimethylthiazolidine-4-
Limit: carboxylic acid (penicilloic acids of ampicillin),
- methylene chloride: maximum 0.2 per cent mlm.
Water (2.5.12) OH~
H',
o
NH'~_}~-
~'~.CH, 0
Bacterial endotoxin. (2.6.14)
Less than 0.15 IU/mg, if intended for use in the manufacture
of parenteral preparations without a further appropriate
procedure for the removal of bacterial endotoxins.
M"'" I
"" 0
-Ht-t-
H
S CH,
E. (2R)-2-[[[(2S,5R,6R)-6-[[(2R)-2-amino-2-phenylacetyl]
ASSAY
Liquid chromatography (2.2.29) as described in the test for amino ]-3,3-dimethyl-7-oxo-4-thia-l-azabicyclo[3.2.0]hept-
related substances with the following modifications. 2-yl]carbonyl]amino]-2-phenylaceric acid (ampiciUinyl-D-
phenylglycine),
Mobile phase Initial composition of the mixture of mobile
phases A and B, adjusted where applicable.
Injection Test solution (a) and reference solution (a).
Calculate the percentage content of ampicillin sodium by
multiplying the percentage content of ampicillin by 1.063. F. (2RS,4S)-2-[[[(2R)-2-amino-2-phenylacetyl]
amino]methyl]-5,5-dimethylthiazolidine-4-earboxylic acid
(penilloic acids of ampicillin),
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2022 Ampicillin Trihydrate 1-181
HNIyC)
NH
crtr
G. (3R,6R)-3,6-diphenylpiperazine-2,5-dione,
N. oligomers of penicilloic acids of ampicillin.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE'"
H. 3-phenylpyrazin-2-<l1. Ampicillin Trihydrate

1. (2S,5R,6R)-6-[[(2R)-2-[[(2R)-2-amino-2-
phenylacetyl] amino] -2-phenylacetyl] amino] -3,3-dimethyl- 403.5 7177-48-2
7-oxo-4-thial-azabicycio[3.2.0]heptane-2-carboxylic acid
(p-phenylglycylampicillin), Action and use
Penicillin antibacterial.
Preparations
Ampic~lin Capsules
Ampicillin Oral Suspension
Co-f1uampicil Capsules
Co-ffuampicil Oral Suspension
J. (2S,5R,6R)-6-[(2,2-dimethylpropanoyl)amino]-3,3-
dimethyl-7-oxo-4-thia-l-azabicycio[3. 2.0] heptane-2- PIlE", _
carboxylic acid,
DEFINITION
(2S,5R,6R)-6-[[(2R)-2-Amino-2-phenylacetyl]amino]-3,3-
dimethyl-7-oxo-4-thia-l-azabicycio[3.2.0]heptane-2-
carboxylic acid trihydrate.
Content
K. (2R)-2-[(2,2-dimethylpropanoyl)amino]-2-phenylacetic CHARACTERS
acid, Appearance
H NH,
«~H
Solubility
Slightly soluble in water, practically insoluble in ethanol
(96 per cent)' and in fatty oils. It dissolves in dilutesolutions
of acids and of alkali hydroxides.
L. (2R)-2-amino-2-phenylacetic acid (D-phenylglycine), IDENTIFICATION
First idenlification: A., D.
Second idemlfication: B., C, D.
Comparison ampiciUin tn'hydrate CRS.
examined in 10 mL of sodium hydrogen carbonate solutUm R.
Reference solution (a) Dissolve 25 mg of ampi<illin
trihydrate CRS in 10 mL of sodium hydrogen carbonate
sduuon R.
J\l.co-oligomers of ampicillin and of penicilloic acids of trihydrate CRS and 25 mg of ampicillin trihydrate CRS in
ampicillin, 10 mL of sodium hydrogen carbonate solution R.
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1-182 Ampicillin Trihydrate 2022
Plate TLC silanised silica gel plate R. - mobile phase B: mix 0.5 mL of dilute aau"c acid R, 50 mL
Mobile phase Mix 10 volumes of acetone Rand 90 volumes of 0.2 M potassium dihydrogen phosphate Rand 400 mL of
of a 154 gIL solutionof ammonium acetate R previously acetonitrile R, then dilute to 1000 mL with water Rj
adjusted to pH 5.0 with glacial acetic acid R.
TUn, MobUe phase A MobUe phase B
Application I ~L. (min) (per cent VIJi) (per cent VIJI)
Development Overa path of 15 em.
o· tR 85 15
Drying In air. tR- (fR + 30) 85 ---> 0 15 -> 100
Detection Expose to iodine vapouruntil the spots appear (IR + 30) - ('R + 45) 0 [00
and examine in daylight. (tR + 45) - (tR + 60) 85 15
System suitabrlity Reference solution (b): tR = retention time of ampicillin determined with reference solution (c)
Results The principal spot in the chromatogram obtained If the mobile phase composition has been adjusted to achieve
with the test solution is similar in position) colour and size to the required resolution) the adjusted composition will apply
the principal spot in the chromatogram obtained with at time zero in the gradient and in the assay.
reference solution (a). Flow TaU 1.0 mf/mlo.
C. Place about 2 mg in a test-tube about 150 mm long and Detection Spectrophotometer at 254 nm.
about 15 mm in diameter. Moisten with 0.05 mL of water R Inje<tion 50 ~ of reference solutions (b) and (c) with
and add 2 mL of sulfuric acid-formaldehyde reagent R. Mix the isocratic elution at the initial mobile phase composition and
contents of the rube by swirling; the solution is practically 50 ~L of test solution (b) according to the elution gradient
colourless. Place the test-tube in a water-bath for 1 min; described underMobile phase; inject mobile phase A as a
a dark yellow colour develops. blank according to the elution gradient described under
D. Water (see Tests). Mobile phase.
TESTS System suitability Reference solution (b):
Appearance of solution - resolution: minimum 3.0 between the peaks due to
The solutions are not more opalescent than reference ampicillin and cefradin; if necessary, adjust me ratio A:B
suspension II (2.2.1). of the mobile phase.
Dissolve 1.0 g in 10 mL of 1 M hydrochloric acid. Separately Limit:
dissolve 1.0 g in 10 mL of drlute ammonia R2. Examine - any impun·ty: for each impurity) not more than the area of
immediately after dissolution. the principal peak in the chromatogram obtained with
pH (2.2.3)
3.5 to 5.5. N,N-D1methylaniline (2.4.26, Melhod B)
Maximum 20 ppm.
Dissolve 0.1 g In carbon dioxide-free water R and dilute to
40 mL with the same solvent. Water (2.5.12)
Specific optical rotatton (2.2.7)
+ 280 to + 305 (anhydrous substance). Sulfured ash (2.4.14)
Dissolve 62.5 mg in water R and dilute to 25.0 mL with the Maximum 0.5 per cent, determined on 1.0 g.
same solvent. ASSAY
Related substances Liquid chromatography (2.2.29) as described in the test for
Liquid chromatography (2.2.29). related substances with the following modifications.
Test solutio,., (a) Dissolve 31.0 mg of the substance to be Mobile phase Initial compositionof the mixture of mobile
examined in mobile phase A and dilute to 50.0 mL with phasesA and B, adjusted where applicable.
mobile phase A. Injection Test solution (a) and reference solution (a).
Testsolution (b) Dissolve 31.0 mg of the substance to be System suitability Reference solution (a):
examined in mobile phase A and dilute to 10.0 mL with - repeatabih'ty: maximum relative standard deviation of
mobile phase A. Prepare immediately before use. 1.0 per cent after 6 injections.
Reference solution (a) Dissolve 27.0 mg of anhydrous Calculate the percentage content of ampicillin from the
ampi<illin CRS in mobile phase A and dilute to 50.0 mL with declared content of anhydrous ampi<illin CRS.
mobile phase A. STORAGE
Reference solution (b) Dissolve 2 mg of cefradine CRS in In an airtight container.
To 5 mL of this solution) add 5 mL of reference solution (a). IMPURITIES
Column:
- size: 1 =. 0.25 m, 0 = 4.6 rom;
- stationary phase: ocradecylsilyl silica gelfor chromatography R
(5 pm).
A. (2S,5R,6R)-6-amino-3,3-dimethyl-7-oxo-4-thia-l-
Mobile phase: azabicyclo[3.2.0]heptane-2-carboxylic acid
- mobile phaseA: mix O.5.mL of dilute acetic acidR, 50 mL (e-amloopenlcillanic acid),
of 0.2 M potassium dihydrogetl phosphate Rand 50 mL of
acetonitrile R, then dilute to 1000 mL with waterR;
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2022 Ampicillin Trihydrate 1-183
B. (2S,5R,6R)-6-[[(2S)-2-amino-2-phenylacetyl]amino]-3,3-
dimethyl-?-oxo-4- thia-l-azabicyclo[3.2.0] heprane-z- I. (2S,5R,6R)-6-[[(2R)-2-[[(2R)-2-amino-2-
carboxylic acid (r-ampicifiln), phenylacetyl]amino]-2-phenylacetyl]amino]-3,3-dimethyl-
?-oxo-4-thia-l-azabicyclo[3.2.0]hep,ane-2-carboxylic acid
(n-phenylglycylampicillin),
C. (4S)-2-(3.6-dioxo-5-phenylpiperazin-2-yI)-5,5-dimethyl- 1. (2S,5R.6R)-6-[(2,2-dimethylpropanoyl)amino]-3,3-
l,3-rhiazolidine-4-cacboxylic acid (diketopiperazmes of dimethyl-?-oxo-4-thia-l-azabicydo[3.2.0]heptane-2-
ampicillin), carboxylic acid,
D. (4S)-2-[[[(2R)-2-amino-2-phenylacetyl]arnino]
K. (2R)-2-[(2,2-dimethylpropanoyl)amino]-2-phenylacetic
carboxymethyl)-5,5-dimethyl-l,3-thiazolidine-4-carboxylic
acid,
acid (penicilloic acids of ampicillin),
L. (2R)-2-amino-2-phenylacetic acid (D-phenylglycine),
E. (2R)-2-[[[(2S,5R,6R)-6-[[(2R)-2-amino-2-
phenylacetyl]amino]-3, 3-dimethyl-?-oxo-4-thia-l-
azabicyclo[3.2.0]hept-2-yl)carbonyl]arnino]-2-phenylacetic
acid (arnpiciUinyl-D-phenylglycine),
F. (2RS,4S)-2-[[[(2R)-2-amino-2-phenylacetyl] .M.co-oligomers of ampicillin and of penicilloic acids of

amino] methyl]-5,5 -dimethyl-l,3-thiazoIidine-4-carboxylic ampicillin,
acid (penilloic acids of ampicillin),
H NH,
~ ('J 0~f~NHCo,H
HN~
H
V goAs 'H
CIi,
(J(irNH CH,
N. (3S)-6-[[(2R)-2-amino-2-phenylacetyl]amino]-2,2-
dimethyl-7-oxo-2,3,4,7-retrahydro-I ,4-rhiazepine-3-
G. (3R,6R)-3,6-diphenyJpiperazine-2,5-dione, carboxylic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PflE<r
H.3-phenylpyrazin-2-ol,
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1-184 Amylmetacresol 2022
- size: I:::: 30 m, 12) = 0.25 mm;

Amylmetacresol - stationary phase: meurogol 20 000 R (film thickness
(Ph. Eur. monograph 2405) 0.5 urn).
H'CYy0H Linear veJodry 33 cm/s.
~CH,
Sp/itratio 1:30.
Temperature:
178.3 1300-94-3 Time Temperature

(min) CC>
Action and use Column 0·17.5 100 ...... 240
Antiseptic. 17.5·32.5 240
Phf" _ Injection port 2'0
Detector 2'0
DEFINITION
5-Methyl-2-pentylphenol. Detection Flame ionisation.
Content Injection 1.0 ....L of test solution (a) and reference
98.0 per cent to 102.0 per cent. solutions (a), (b) and (d).
CHARACTERS Identification of impurities Use the chromatogram supplied
Appearance with amylmeuuresol for peak idemijication CRS and the
Clear or almostclear liquid, or solid crystalline mass, chromatogram obtained with reference solution (b) to
colourless or slightly yellow when freshly prepared. identify the peaks due to impurities A, G and K.
The substance changescolour during storage by darkening Relativemention With reference to amylmetacresol
andlordiscolouration to dark yellow) brownish-yellow or (retention time:::: about 16 min): impurity G
pink. (diastereoisomer 1) = about 0.51; impurity G
Solubility (diastereoisorner 2) = about 053; impurity D = about 0.77;
Practically insoluble in water, very soluble in acetone and in impurity B = about 0.78; impurity K = about 0.95;
ethanol (96 per cent). impurity A = about 0.99.
It solidifies at about 22°C. System suitability Reference solution (a):
IDENTIFICATION impurities D and B.
Limits:
Preparation Film between 2 plates of potassium bromide R. - impurity A: maximum 0.6 per cent;
Comparison amy/metacresol CRS. - impurities G (sum of the 2 diastereoisomers), K: for each
TESTS impurity, maximum 0.15 per cent;
Related substances - unspe.cified impurities: for each impurity, maximum
Gas chromatography (2.2.28): use the normalisation 0.10 per cent;
procedure. - total: maximum 1.0 per cent;
- disregard limit: the area of the peak due to amylmetacrescl
Internal standard solution Dissolve 0.100 g of
butylhydroxytoluene R in 2-propanol R and dilute to 10.0 rnL
(0.05 per cent).
Sulfated ash (2.4. 14)
examined in 2-propanol R and dilute to 10.0 rnL with the
same solvent. ASSAY
Test solution (b) To 2.0 mL of test solution (a) add 2.0 rnL Gas chromatography (2.2.28) as described in the test for
of the internal standard solution and dilute to 10.0 mL with related substances with the following modification.
2-propanol R. Injection 1.0 IAL of test solution (b) and reference
Reference solution (a) Dissolve 10 mg of m-cresd R solution (c).
(impurity B) and 10 mg of p-eresoI R (impurity D) in Calculate the percentage content ofC1ZH1SO from the
2-propanol R and dilute to 100.0 mL with the same solvent. declared content of amylmeuu:resol CRS.
Reference solutIOn (b) Dissolve the contents of a vial of STORAGE
amybneuuresol for peak idendfication CRS (containing In an airtight, non-metallic container, protected from light.
impurities A, G and K) in 1.0 rnL of 2-propanol R.
Reference solution (r) Dissolve 0.1 000 g of
IMPURITffiS
amylmetacresol CRS in 2-propanol R and dilute to 10.0 mL Specified impurities A, G, K.
with the same solvent. To 2.0 mL of this solution add Otherdew;rable impurities (thefollowing substances would, if
2.0 mL of the internal standard solutionand dilute to present at a sufficient levd, be detected by oneor other of the tests
10.0 rnL with 2-propanol R. in the monograph. They are limited by the general aaeptante
Reference solution (d) Dilute 1.0 rnL of test solution (a) to criterion for other/unspecified impurities and/or by the general
100.0 rnL with 2-propanol R. Dilute 1.0 rnL of this solution
monograph Substances for phannaceutital use (2034). It is
to 20.0 rnL with 2-propanol R.
therefore not nuessary to identify these impun"ties for
demonstration ofromp/iane<. See also 5.10. Control of impurities
Column: it, substances for phannaceutical use) B, G, D, E, F, H, I, J.
- material: fused sillca;
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2022 Anastrozole 1-185
Anastrozole
A. 4-methyl-2-pentylphenol,
B. 3-methylphenol (m-cresol),
H 293.4 120511-73-/
H'C'OC5c
?" I H., CHa and enantiomer
~ CHa
Action and use
Aromotase inhibitor; treatment of breast carcinoma.
C.5-methyl-2-[(2RS)-2-methylbutyljphenol, Preparation
Anastrozole Tablets
!""'y0H PhE" ~ ~ _
H3C~ DEFINITION
2,2 '-[5-( I H-I ,2,4-T riazol-I-ylmethyl) benzene-I,3-diyl)bis(2-
D. 4-methylphenol (p-cresol), methylpropanenitrile).
Content
CHARACTERS
Appearance
E. 1-(2-hydroxy-4-methylphenyl)pentan-I-one, Solubility
Very slightly soluble in water, freely soluble in anhydrous
ethanol, practically insoluble in cyclohexane.
IDENTIFICATION
F. 1-(2-hydroxy-5-methylphenyl)pentan-I-one, Comparison anastrozok CRS.
substance separately in acetone R) evaporate [0 dryness and
recordnew spectra using the residues.
TESTS
G.5-methyl-2-pentylcyclohexanone, Related substances
Solvent mixture acetonitrile Rl, waterfor chromatography R
(50:50 VIP).
Test solutUm (a) Dissolve 25 mg of the substance to be
H. ethyl pentanoate, examined in. the solvent mixture and dilute to 100.0 mL with
H'CUO~
I I CH, Test solution (b) Dissolve 25.0 mg of the substance to be
"" 0
I. 3-methylphenyl pentanoate, Reference sdtuion (a) Dilute 1.0 mL of test solution (a) to
Referen" solution (1)) Dissolve 2.5 mg of anastrozo/e
impurity E CRS in 20.0 mL of the solvent mixture. Dilute
1.0 mL of the solution to 50.0 mL with test solution (a).
J. 4-methyJphenyl pentanoate, Reference solution (c) Dissolve 25.0 mg of anastrozole CRS in
K. unknown structure. the solvent mixture and dilute to 200.0 mL with the solvent
_~ PhE" mixture.
Column:
- size: 1= 0.15 m, 0 = 4.6 mm;
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1-186 Anastrozole 2022
- stationary phase: end-capped ethylene-bridged polar-embedded

octade<ylsilyl silica gelfor chromatography (hybrid materia/,! R
(3.5 pm).
Mobile phase:
- mobile phase A: phosphon'c acid R, waterfor
chromatography R (0.1:100 V/II);
- mobile phase B: phosphoric acid R, acewnitrile Rt A. 2-[3-[(lRS)-I-cyanoethyl]-5-(lH-I,2,4-triazol-l-ylmethyl)
(0.1:100 V/II); phenyl)-2-methylpropanenitrile,
Thn. MobUe phase A Mobile phase B

(min) (per cent VIJ.') (per cent VjJJ)
0-2 95 5
2 - 54 95 -> 35 5 ..... 65
Flow rate 1.0 mIlmin.

Detection Spectrophotometer at 215 nm. H,C andenantiomef
Injection 20 ilL of test solution (a) and reference CN
H,C
Identification of impurirres Use the chromatogram obtained B. (2RS)-2,3-bis[3-(I-cyano-I-methylethyl)-5-(IH-I,2,4-
with reference solution (b) to identify the peak due to triazol-I-ylmethyl) phenyll-2-methylpropanenitrile,
impurity E.
Relau"ve retention With reference to anastroaole (retention
time = about 29 min): impurity E = about 1.05.
- resolurion: minimum 3.5 between the peaks due to
anastrozole and impurity E.
- for each impurity, use the concentration of anastrozole in C. 2,2'-[5-(bromomethyl)ben2ene-1,3-diyl] bis(2-
reference solution (a). methylpropanenitrile),
Limits:
- unspeaJied {mpuniies: for each impurity, maximwn
0.10 per cent;
Water (2.5.32)
Maximum 0.3 per cent, determined on 50.0 mg.
D.2,2'-[5-(dibromomethyl)benzene-I,3-diyl]bis(2-
Maximum 0.1 per cent) determined on 1.0 g.
methylpropanenitrile),
ASSAY
Injection Test solution (b) and reference solution (c).
Calculate the percentage content of C l1H19N j taking into
account the assigned content of anastrozole CRS"
IMPURITIES
Otherdetectable impurities (the following substances would, if E. 2,2'-[5-(hydroxymethyl)benzene-l,3-diyllbis(2-
present at asufficknt leoel, be detected by oneor otherof the tests methylpropanenitrile),
in the monograph. They are limited by thegeneral accepumce
monograph Substances for phannacenticalnse (2034). It is
demonstration of compliance. See also 5.10. Control of t"mpuniies
in substances for pharmaceutical use) A, B) C, DJ EJ F, G, F. 4-methylbenzenesulfonic acid,
H) 1.
G. 2,2'-[5-(4H-I,2,4-triazol-4-ylmethyl)ben2ene-I,3-diyllbis
(2-methylpropanenitrile),
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2022 Animal Epithelia and Outgrowths for Allergen Products 1-187
Where major changes to the production of the animal

epithelia and outgrowths take place (e.g. when a new process
or supplier is introduced), such changes are qualified.
Microbial contamination of the animal epithelia and
outgrowths may be unavoidable and should be monitored on
a representative number of batches of source material
according to a justified sampling plan and each time a new
H. 2,2'-(5-methylbenzene-I,3-diyl)bis(2- supplier and/or a new process for the source material
methylpropanenitrile), production is introduced; if a determination of microbial
contamination is not applicable, this must be justified.
Microbial contamination values and potential increases in
microbial contamination are monitored during stability
studies, in order to assess this aspect along with the source
material characteristics upon storage.
Control methods and acceptance criteria relating to identity
and purity of the animal epithelia and outgrowths are
established. The acceptance criteria must ensure the
I. 2,2'-[5-(chloromethyl)benzene-I,3-diyl)bis(2- consistency of the animal epithelia and outgrowths source
methylpropanenitrile). material from a qualitative and quantitative point of view.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph,.. The animal epithelia and outgrowths source material is
stored under controUed conditions justified by stability data.
The collection and production, as well as the handling of the
source material, are such that consistent composition is
ensured from batch to batch.
Animal Epithelia and Outgrowths ANIMAL EPITHELIA AND OUTGROWTHS FOR
for Allergen Products ALLERGEN PRODUCTS REFERENCE BATCH
(Ph. Eur. monograph 2621) An appropriate reference batch is established for each animal
epithelia and outgrowths source material. The nature of the
Ph,.. _
reference batch depends on the testing approach to verify
DEFINITION batch-to-batch consistency and to establish acceptable
Animal epithelia and outgrowths for allergen products consist quality. The reference batch may be, for example, an internal
of hair, epithelium fragments, dander, feathers and other reference preparation (if available), a source material extract
structures that grow from the epidermis of mammals or or a sample of a production batch. Its characterisation must
birds. be described. The extent of characterisation of the reference
batch depends on the nature of the animal epithelia and
Animal epithelia and outgrowths may contain proteins
outgrowths source material, knowledge of the allergenic
deposited from the saliva and/or secretions from the
components and availability of suitable reagents.
sebaceous glands of the animal. They may be further
The reference batch is stored under controlled conditions
processed (e.g. cut or washed) using qualified methods or are
ensuring its stability.
unprocessed.
BATCH-TO-BATCH CONSISTENCY
PRODUCTION To establish batch-to-batch consistency, one or more of the
Animal epithelia and outgrowths for allergen products are
following tests are performed on each batch. The choice of
obtained from healthy animals selected to avoid possible tests must be justified.
transmissible agents of disease. The exact species and/or
variety of animal is Slated. Typical production steps, Total protein (2.5.33)
including animal management, source material collection and Protein profile
purification, are specified. The origin, quality, and Determined by using suitable electrophoresis methods
traceability of the source material must be demonstrated. (2.2.31,2.2.54).
It is expected that, where applicable, the animal care and Allergen profile
husbandry follows the principles described for the protection Relevant allergenic components are identified by means of
of vertebrate animals used for experimental and other suitable teclmiques using allergen-specific antibodies.
scientific purposes. A responsible veterinarian or another Major allergen content
competent person confirms the identity of the species and Determined by using suitable inununochemical methods
that the animals are healthy. It is verified that the skin is (2.7.1) such as enzyme-linked immunosorbent assay
visibly clean and intact before harvest and that the animals (EllSA).
have not been recently treated with preparations for
cutaneous application, such as antiparasitic drugs. Total allergenic activity
Determined by testing inhibition of the binding capacity of
The collection of animal epithelia and outgrowths must be
specific immunoglobulin E antibodies or by a suitable
performed without injuring the skin of the animal.
equivalent in vitro method.
Confirmation that measures are in place to prevent cross-
contamination by animal epithelia and outgrowths from other CHARACTERS
animals is provided, including during animal management, Animal epithelia and outgrowths for allergen products are
collection and processing. Methods involving the grinding of supplied as coloured powders or other materials such as
whole skin and/or pelts must not be used. feathers, dander or hairs.
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1-188 Antazoline Hydrochloride 2022
IDENTIFICATION antasoline hydrochloride CRS. Examine the substances as discs

The identity of animal epithelia and outgrowths is confirmed prepared using potassium chloride R.
by their relevant macroscopic and microscopic characteristics B. Examine the chromatograms obtained in the test for
in comparison to those of a reference batch or reference related substances in daylight after spraying, The principal
documents. Identity may also be confirmed using other spot in the chromatogram obtained with test solution (b) is
methods such as EUSA or by genetic identification, if similar in position, colour and size to the principal spot in
performed by generally accepted methods. the chromatogram obtained with reference solution (b).
TESTS C. To 5 mL of solution S (see Tests) add, drop by drop,
Foreign matter dilutesodium hydroxide solution R until an alkaline reaction is
Foreign matter is defined as vermin (e.g. mites and fleas), produced. Filter. The precipitate, washed with two
dirt) and foreign animal epithelia and outgrowths. Foreign quantities, each of 10 ml., of water R and dried in a
matter is determined by appropriate tests (e.g. microscopic desiccator under reduced pressure, melts (2.2. Jtf) at 119 °C
examination, EliSA), visual inspection and/or tactile to 123"C.
inspection. Foreign matter is below a predefined and justified D. It gives reaction (a) of chlorides (2.3.1).
limit.
TESTS
Water (2.5.12 or 2.5.32) or loss on drying (2.2.32) Solution S
The water content of dried material is determined; Dissolve 2.0 g in carbon dioxide-free water R prepared from
specification limits must be supported by batch analysis and distiUed waterR, heating at 60 °C if necessary. Allow to cool
stability data. and dilute to 100 mL with the same solvent.
STORAGE Appearance of solution
The source materials are stored under controlled conditions Solution S is clear (2.2.1) and not more intensely coloured
justified by stability dara. than reference solution Y7 (2.2.2, Method /1).
LABELLING Acidlty or alkalinity
The label states: To 10 mL of solution S add 0.2 mL of methyl red solution R.
- the species of the source animal; Not more than 0.1 mL of 0.01 M hydro<hloric acid or 0.01 M
- the nature of the animal epithelia and outgrowths. sodium hydroxide is required to change the colour of the
_~ PIIE" indicator.
Related substances
Examine by thin-layer chromatography (2.2.27), using silica
gel GF254 R as the coating substance. Heat the plate at
Antazoline Hydrochloride 110 °C for 15 "min before using.
(Ph. Eur. monograph 0972) examined in methanol R and dilute to 5 mL with the same
solvent.
Test sdution (b) Dilute 1 mLofresrsolution (a) to 5 mL
with methanol R.
'HCI Reference solution (a) Dilute 0.5 mL of rest solution (a) to
Reference solution (b) Dissolve 20 mg of antazoline
same solvent.
301.8 2508-72-7
Reference solution (c) Dissolve 20 mg of xy/ometazoline
Action and use hydrochloride CRS in I mL of test solution (a) and dilute to
Histamine Hi receptor antagonist; antihistamine. 5 mL with methanol R.
Apply to the plate 5 pL of each solution. Develop over a
PIlE" _
path of IS em using a mixture of 5 volumes of
DEFINITION diethylamine R, "10 volumes of methanol R and 85 volumes of
Antazoline hydrochloride contains not less than 99.0 per cent ethylacetate R. Dry the plate in a current of warm air for
and not more than the equivalent of 101.0 per cent of 15 min. Examine in ultraviolet light at 254 run. The test is
N-benzyl-N-[(4,5-dihydro-1H-imidazol-2-yl)methyllaniline not valid unless the chromatogram obtained with reference
hydrochloride, calculated with reference to the dried solution (c) shows two clearly separated principal spots.
substance. Spray with a mixture of equal volumes of a 200 gIL solution
of fenic chloride R and a 5 gIL solution of potassium
CHARACTERS
ferneyanide R. Examine immediately in daylighr. Any spor in
A white or almost white, crystalline powder, sparingly soluble the chromatogram obtained with test solution (a), apart from
in water, soluble in alcohol, slightly soluble in methylene the principal spot, is not more intense than the spot in the
chloride. chromatogram obtained with reference solution (a)
It melts at about 240 °C, with decomposition. (0.5 per cent),
IDENTIFICATION Loss on drying (2.2.32)
First idendfication: A, D. Not more than 0.5 per cent, detennined on 1.000 g by
Secondidentification: B, C, D. drying in an oven at 105 °C for 3 h.
A. Examine by infrared absorption spectrophotometry
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2022 Apomorphine Hydrochloride Hemihydrate 1-189
Sulfated ash (2.4.14) solution. Dilute J0.0 mL of the solution to 100.0 mL with a
Not more than 0.1 per cent, determined on me residue 10.3 gIL solution of hydrochloric acid R.
obtained in the test for loss on drying. Spectral range 230-350 om
ASSAY Absorption maximum At 273 run.
Dissolve 0.250 gin 100 mL of akohol R. Add 0.1 mL of Shoulder At 300-310 nm.
phenolph,hakin solution RJ. Titrate with 0.1 M alroholit Specific absorbana. at the absorption maximum 530 to 570.
potassium hydroxide.
I mL of 0.1 M akoholitpotassium hydroxide is equivalent to
Comporison apomorphine hydrochloride hemihydrate CRS.
30.18 mg of C 17H,oCIN,.
C. To 5 mL of solution S (see Tests) add a few millilitres of
IMPURITIES sodium hydrogen carbonate solution R until a permanent, white
precipitate is formed. The precipitate slowly becomes
greenish. Add 0.25 mL of 0.05 M iodine and shake.
The precipitate becomes greyish-green. Collect the
precipitate. The precipitate dissolves in methylene chloride R
giving 3 violet-blue solution and in ethanol (96 per cent) R
giving a blue solution.
D. To 2 mL of solution S (see Tests) add 0.1 mL of nitric
A. N-(2-aminoethyl)-2-(benzylphenylamino)acetamide. acidR. Mix and filter. The filtrate gives reaction (a) of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE" chlorides (2.3.1).
TESTS
Solution S
Dissolve 0.25 g without heating in carbon dioxide-free waterR
Apomorphine Hydrochloride *** and dilute [0 25 mL with the same solvent.
*** *** Appearance of soludon
Hemihydrate *** Solution S is clear (2.2.1) and not more intensely coloured
(Ph. Eur. monograph 0136) than reference solution BY, or GY j (2.2.2, Method If).
pH (2.2.3)
4.0 to 5.0 for solution S.
Ho@OH
· :
. Hel . '/ 2 H20
1 -52 to -48 (dried substance).
"'" H,CH,
N
Dissolve 0.25 g in a 2.06 gIL solution of hydrochloric acid R
and dilute to 25.0 mL with the same acid solution.
Related substances
312.8 41372-20-7 liquid chromatography (2.2.29).
Action and use Test solution Dissolve 50.0 mg of the substance [0 be
Dopamine receptor agonist; treatment of Parkinson's disease. examined in a 1 per cent VIV solution of glacial acetic acid R
and dilute to 20.0 mL with the same solution.
Preparation
Apomorphine Hydrochloride for Homoeopathic Preparations Reference solution (a) Dilute 1.0 mL of the test solution to
100.0 mL with a 1 per cent VIV solution of gla<ial acetic
PIlE" _ acid R. Dilute 1.0 mL of this solution to JO.O mL with a
DEFINITION 1 per cent VIV solution of glacial acetic acid R.
(6aR) -6-Methyl-5,6,6a, 7-tetrahydro-4H-dibenzo[de,gJ Reference solution (b) Dissolve 12.5 mg of apomorphine
quinoline-IO,II-diol hydrochloride hemihydrate. impurity B CRS in a 1 per cent VI V solution of glacial acetic
acid R and dilute to 10.0 mL with me same solution.
Content
98.5 per cent to 101.5 per cent (dried substance). Reference solution (c) Dilute 2.0 mL of reference solution (b)
to 10.0 mL with a 1 per cent VIV solution of glacial acetic
CHARACTERS acidR. Dilute 2.0 mL of this solution (0 100.0 mL with a
Appearance 1 per cent VIV solution of glacial acetic acid R.
White or slightly yellowish-brown or green-tinged greyish, Reference solution (d) Dissolve 25 mg of boldine R in a
crystalline powderor crystals; on exposure to air and light, 1 per cent VIV solution of gku:ial acetic acid R and dilute to
the green tinge becomes more pronounced. 10 mL with the same solution. To I mL of this solution add
Solubility 1 mL of me test solution and dilute to 10 mL with a
Sparingly soluble in water and in ethanol (96 per cent), I per cent VIV solution of gkuial acetic acidR.
practically insoluble in toluene. Column:
IDENTIFICATION - size: 1= 0.15 m, 0 = 4.6 mm;
First identification: B, D. - stationary phase: end-capped oetadecylsilyl silica gelfor
Second identification: A, C, D. chromatography R (5 urn);
(2.2.25). Mobile phase:
- mobile phaseA: 1.1 gIL solution of sodium
Test solution Dissolve 10.0 mg in a 10.3 gIL solution of octanesulfonate R} adjusted to pH 2.2 with a
hydrochloric acid R and dilute to 100.0 mL with the same acid 50 per cent mlm solution of phosphonc acidR;
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1-190 Aprepitant 2022
- mobile phase B: acetonitrile R;

(min) (per cent VIJI) (per cent VIJ')
0-2 85 15
2·32 85 ---> 68 15 ---> 32
32 - 37 68 32
A. (6aRJ-IO-methoxy-6-methyl-5,6,6a,7-tetrahydro-4H-
dibenzo[de,g]quinolin-ll-ol (apocodeine),
Flow rau 1.5 mUmin.
Detection Spectrophotometer at 280 om. ,CH 3
~
H\ N
lnjeaion 10 1'1-.
Identification of impun"ties Use the chromatogram obtained _ H
with reference solution (b) (Q identify the peak due to
impurity B. ..... ! H
Relative retention With reference to apomorphine (retention HO 0HOH
=
time about 18 min): impurity B =about 0.4;
boldine = about 0.9. B. 7,8-didehydro-4,5ot-epoxy-17-methylmorphlnan-3,6ot-diol
Systemsuitability Reference solution (d): (morphine),
- resolution: minimum 2.5 between the peaks due to boldine
and apomorphine.
Calculation of percentage contents.:
- for impurity B, use the concentration of impurity B in
reference solution (e);
- for impurities other than B, use the concentration of
apomorphine hydrochloride hemihydrate in reference
solution (a).
Limits:
- impurity B: maximum 0.15 per cent; C. (6aR)-9-[7,8-didehydro-4,5ot-epoxy-3-hydroxy-17-
- unspecified impurities: for each impurity, maximum methyhnorphinan-6ot-yl)-6-methyl-5,6,6a,7-tetrahydro-4H-
0.10 per cent; dibenzo[de,g]quinoline-I 0, ll-diol (morphine-apomorphine
- toUJ1: maximum 0.5 per cent; dimer).
- reporting threshold: 0.05 per cent. ___________________ Ph,,,
drying in an oven at 105°C for 2 h.
Sulfated ash (2.4.14) Aprepitant
ASSAY
Dissolve 0.250 g in a mixture of 5.0 mL of 0.01 M
hydroch/orn acid and 50 mL of ethanol (96 per cenQ R. Carry
out a potentiometric titration (2.2.20), using 0.1 M sodium
hydroxide. Read the volume added between the first 2 points
of inflexion.
C 17H,.CINOa.
STORAGE
IMPURITIES 534.4 170729-80-3
Specified impurities B. Action and use
Otherdetectable impuri'ies (the following substances would, if Neurokinin-l (NKt) receptor antagonist; prevention of
present at a sufficient level, be detected by oneor other of the tests nausea and vomiting associated with emetogenic
in the monograph. They are limited by the general acceptance chemotherapy.
cruenon for other/unspecified impurities and/or by thegeneral Preparation
monograph Substances for pharmaceutical use (2034).1, is
Aprepitant Capsules
therefore not neassary to identify these impuniies for
demonstration of compliance. See also 5.10. Control of impurities Ph'" _
in substances for pharmaceutical use) A, C.
DEFINITION
5-[[(2R,3S)-2-[( I RJ-1- [3, 5-Bis(trilluoromethyl)
phenyl]ethoxy]-3-(4-lluorophenyl)morpholin-4-yl]methyl]-
I,2-dihydro-3H-I ,2,4-triazol-3-one.
Content
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2022 Aprepitanr 1-191
CHARACTERS chromatogram obtained with reference solution (c) to identify

Appearance the peak due to impurity A.
White or almost white powder. Relative retention With reference to aprepitant (retention
Solubility time = about 15 min): impurity A = about 0.97.
Veryslightly soluble in water, sparingly soluble in anhydrous System suitability Reference solution (c):
ethanol, praeticaUy insoluble in heptane. - resolution: minimum 1.5 between the peaks due to
It shows polymorphism (5.9). impurity A and aprepitanr.
IDENTIFICATION Limits:
A. Specific optical rotation (see Tests). - impun'tyA: maximum 0.15 per cent;
- unsP«ified impurities: for each impurity, maximum
B. Infrared absorption spectrophotometry (2.2.24). 0,10 per cent;
Comparison aprepitant CRS. - total: maximum 0.2 per cent;
If the spectra obtained in the solid stateshow differences, - reporting threshold: 0.05 per cent (reference solution (b)).
dissolve the substance to be examined and the reference Water (2.5.32)
substance separately in anhydrous ethanol R, evaporate to Maximum 0.2 per cent, determined on 0.200 g by direct
dryness on a water-bath and record new spectra using the sampleintroduction.
residues.
TESTS Maximum 0,1 per cent, determined on 1.0 g in a platinum
Specific optical rotation (2.2.7) crucible.
+ 66.0 to + 70.0 (anhydrous substance), measured at 25 ·C.
ASSAY
Dissolve 0.250 g in methanolR and dilute to 25.0 mL with Liquid chromatography (2.2.29) as described in the test for
me same solvent. related substances with the following modification.
Related substances Injection Test solution and reference solution (a).
Liquid chromatography (2.2.29): use the nonnaJisation
Calculate the percentage content of C2~21F1N403 taking
procedure.
into account the assigned content of aprepium: CRS.
Solvent mixture acetonitrile Rl, waterfor chromatography R
(50:50 VII'). IMPURITIES
Spedfied impurities A.
Test solution Dissolve 40.0 mg of the substance to he
examined In the solvent.mixture and dilute to 50.0 mL with Otherdetectable impurities (the folWwing substances would, if
the solvent mixture. present at a rufficient /evel~ be detected by one or other of the tests
in the monograph. They are limited /ry the general acceptance
Reference solution (a) Dissolve 40.0 mg of aprepitant CRS in
cruerion for other/unspecified impun'lies and/or by the general
the solvent mixture and dilute to 50.0 mL with the solvent
monograph Substances for phannaautical use (2034). It is
mixture.
therefore not n«essary to identify these impuniies for
Reference solution (b) Dilute 1.0 mL of the test solution to demonstration of compliance. See also5.10. Control of ;mpw;ties
100.0 mL with the solvent mixture. Dilute 1.0 mL of this in substances for pharmaceutical use) B~ C.
solution to 20.0 mL with the solventmixture.
Reference solution (c) Dissolve 4 mg of aprepiUlnt for system
suitability CRS (containing impurity A) in 5.0 mL of the
solvent mixture.
Column:
- size: I ;;;; 0.25 ill, 0 ;;;; 4.6 mm;
- stationary phase: end-capped octadecylsilyl silica gelfor
chromatography compatible with lOOpercentaqueous mobile
phases R (5 urn);
- temperature: 35°C. A. 5-[[(2R,3S)-2-[ (I R)-I-[3,5-bis(trifluoromethyl)
Mobilephase: phenyl]ethoxyj-3-phenylmorpholin-4-yl)methylj-I,2-
- mobile phaseA: 0.1 per cent VIV solution of phosphoric dihydro- 3H-I ,2,4-ttiazol-3-one,
acid R;
- mobile phase B: acetonitrile R1;
Thne MobIle phase A MobIle phase B

(min) (per cent VIJI) (per cent VII?
0-2 65 35
2 - 22 65 --> 20 35 --> 80
22 - 32 20 80
B. 5- [[ (2R, 3S)-2-[(1R) -1-[3,5-bis(trifluoromethyl)

Flow rate 1.5 mUmin. phenyljethoxyj -3-(4'-fiuorobiphenyl-3-yl)morpholin-4-
Detection Spectrophotometer at 220 om. yl]methyl]-1,2-dihydro-3H-l,2,4-triazol-3-one,
Injection 20 JJL of the test solution and reference
solutions (b) and (c).
Identification of impuniies Use the chromatogram supplied
with aprepitant for system suitaln7iry CRS and the
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1-192 Aprotinin 2022
Plate TLC silica gel G plate R.

Mobile phase water R, glacial actticacid R (80:100 VIV)
containing 100 gIL of sodium acetate R.
Applicalion 10 ~L.
Drying In air.
Deuaion Spraywith a solution of 0.1 g of ninhydnn R in a
mixture of 6 mL of a 10 gIL solution of cupric chloride R,
21 mL of glacial acetic acid Rand 70 mL of anhydrous
emanol R. Dry the plate at 60°C.
C. 5-[ [(2R,3S)-2-[(I R)-I-[3,5-bis(triftuoromethyl)
Resulu The principal spot in the chromatogram obtained
phenyl)ethoxyl-3-(4'-ftuorobiphenyl-4-yl)morpholin-4- with the test solution is similar in position, colourand size to
yljmethylj-I ,2-dihydro-3H-I,2,4-rriazol-3-one.
the principal spot in the chromatogram obtained with the
_____________________ """ reference solution.
B. Determine the ability of the substance to be examined to
inhibit trypsin activity using the method described below.
Test solution Dilute 1 mL of solution S to 50 mL with bll/fer
solulion pH 7.2 R.
Aprotinin Trypsin sdution Dissolve 10 mg of lrypsin BRP in 0.002 M
(Ph. Eur. monograph 0580) hydrochloric acid and dilute to 100 mL with the same acid.
Casein solution Dissolve 0.2 g of casein R in buffer solution
H- Arg- Pro- Asp- Phe- Cys-Leu - Glu- Pro- Pro- Tyr -
pH 7.2 R and dilute to 100 mL with the same buffer
Thr-Gly-Pro-Cys-Lys-Ala-Arg-lIe-lle-Arg- " solution.
Tyr-Phe-Tyr-Asn Ala Lys Ala GI, Leu "
Cys- Precipitating solution glacial acetic acidR, water R, anhydrous
"AIa- ethanol R (1:49:50 VIVIV).
GO> TlV'-Phe Va' T" G~ G~ Cys-Arg
.. Mix 1 mL of the test solution with 1 mL of the trypsin
Lys- Arg- Asn- Asn-Pbe- Lys- ser-: Ala Glu
,.
Asp-
solution. Allow to stand for 10 min and add 1 mL of the
Cys-Mel- Arg- Thr-cye - Gly-Gfy-A1a-OH casein solution. Incubateat 35°C for 30 min. Cool in iced
" waterand add 0.5 mL of the precipitating solution. Shake
and allow to stand at room temperature for 15 min.
6511 9087-70-1 The solution is cloudy. Carry out a blank test under the
same conditions using bufftr solution pH 7.2 R instead of the
Action and use test solution. The solution is not cloudy.
Amilibrinolytic.
TESTS
""" - - - - - - - - - - - - - - - - - - - - - Solutiun S
DEFINITION Prepare a solution of the substance to be examined
Aprotinin is a polypeptide consistingof a chain of 58 amino containing 15 Ph. Eur. U.lmL, calculated from the activity
acids. It inhibits stoichiometrically the activity of several stated on the label.
proteolytic enzymes such as chymotrypsin, kallikrein, plasmin Appearance of solution
and trypsin. It contains not less than 3.0 Ph. Eur. U. Solution S is clear (2.2.1).
of aprotinin activity per milligram, calculated with reference Absorbance (2.2.25)
to the dried substance. Maximum 0.80 by measuring at the absorption maximum at
PRODUCTION 277 nm.
The animals from which aprotinin is derived must fulfil the Prepare a solution of the substance to be examined
requirements for the health of animals suitable for human containing 3.0 Ph. Eur. U.lmL.
consumption. Des-Ala-aprotinln and des-Ala-des-Gly-aprotlnln
The method of manufacture is validated to demonstrate that Capillary zone electrophoresis (2.2.47): use the normalisation
the product) if tested, would comply with the following test. procedure.
Histamine (2.6.111) Testsolution Prepare a solution of the substance to be
Maximum 0.2 ug of histamine base per 3 Ph. Eur. U. examined in water R containing not less than
CHARACTERS 1 Pb. Eur. U.lmL.
Appearance Reference solution Dilute aprou"nin solution BRP in water R to
Almost whitehygroscopic powder. obtain the same concentration as the test solution.
Solublllty Capalary:
Soluble in water and in isotonic solutions, practically - material: uncoated fused silica;
insoluble in organic solvents. - size: effective length = 45-60 em, 0 = 75 JAm.
Temperature 25°C.
IDENTIFICATION
A. Thin-layer chromatography (2.2.27). CZE bll/fer Dissolve 8.21 g of potassium dihydrogen
phosphate R in 400 mL of waterR, adjust to pH 3.0 with
Test solution SolutionS (see Tests).
phosphoric acid R, dilute to 500.0 mL with waterR and filter
Reference solulIlm Dilute aprotinin solution BRP in waterR to through a membrane filter (nominal pore size 0.45 um),
obtaina concentration of 15 Ph. Eur. UJmL.
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2022 Aprotinin 1-193
Detection Spectrophotometer at 214 run. Relative retention With reference to aprotinin (retention
Between-run rinsing Rinse the capillary for at least 1 min time =17.0 min to 20.0 min): impurity C =about 0.9.
with 0.1 M sodium hydroxide filtered through a membrane SYSMm suitilbility Reference solution:
filter (nominal pore size 0.45 pm) and for 2 min wil.h the - resolution: minimum 1.5 between the peaks due 10
CZE buffer. impurity C and aprotinin;
Injection Under pressure or vacuum (for example, 3 s at a - symmetry factor: maximum 1.3 for the peak due to
differential pressure of 3.5 kPa). aprotinin.
Migration Apply a field strength of 0.2 kVlcm, using the Limits:
CZE buffer as the electrolyte in both buffer reservoirs. - impun'ry C: maximum 1.0 per cent;
Run time 30 min. - any other impurity: maximum 0.5 per cent;
- sum of impun"ties other than C: maximum 1.0 per cent.
Identification of impuruies Use the eleetropherogram supplied
with aprotinin solution BRP and the electropherogram Aprotinin ollgomers
obtained with the reference solution to identify the peaks due Size-exclusion chromatography (2.2.30): use the
to impurities A and B. nonnalisation procedure.
Relative migration Wirh reference to aprotinin (migration Test solution Prepare a solution of the substance to be
=
time about 22 min): impurity A :::: about 0.98; ~xamined in waterR containing about 5 Ph. Eur, U JrnL.
impurity B = about 0.99. Reference solution Treat the substance to be examined to
System suitability Reference solution after at least obtain about 2 per cent aprotinin oligomers. For example,
6 injections: heat freeze-dried aprotinin at about 110°C for about 4 h.
- migration time: aprotinin = 19.0 min to 25.0 mini Then dissolve in waterR to obtain a concentration of about
- resolution: minimum 0.8 between the peaks due to 5 Ph. Eur. U.lmL.
impurities A and H; minimum 0.5 between the peaks due Column 3 columns coupled in series:
to impurity Band aprotinin; - size: I;;;; 0.30 m, 0 ;;;; 7.8 mm;
- peak distribution: the electropherogram obtained is - stationary phase: hydrophilic silica gelfor chramatography R
qualitatively and quantitatively similar to the of a grade suitable for fractionation of globular proteins in
electropherogram supplied with aprotini'n sdution BRP; the relative molecular mass range of 20 000 to
- heigh, ofthe principal peak: at least 1000 times the heigbt 10000000 (8 urn).
of the baseline noise. If necessary, adjust the sample load Mobile phase acetonitrile R, glacial acetic acid R, waterfor
to give peaks of sufficient height" chromatography R (2:2:6 VlVlfI); tiller and degas.
Limits: Flow rare 1.0 mUmin.
- impun·ty A: maximum 8.0 per cent; Detection Spectrophotometer at 277 nm,
- impurity B: maximum 7.5 per cent.
Injection 100 [1L.
Pyroglutamyl-aprotinin and related compounds Run time 40 min,
Liquid chromatography (2.2.29): use the normalisation
procedure. Relative retention With reference to aprotinin monomer
(retention time ;;;; 24.5 min to 25.5 min): aprotinin
Test solution Prepare a solution of the substance to be dimer e about 0.9.
examined in mobile phase A, containing about
5 Ph. Eur. U JmL. System suitabilizy Reference solution:
Reference solution Dissolve the contents of a vial of aprotinin
aprotinin dimer and monomer;
for system suilability CRS in 2.0 mL of mobile phase A. - symmetry factor: maximum 2.5 for the peak due to
Column: aprotinin monomer.
- size: I;;;; 0.075 m, 0 ;;;; 7.5 mm; Limit;
- stationary phase: strong cation-exchange silica gelfor - lotal: maximum 1.0 per cent.
- temperature: 40°C. Loss on drying (2.2.32)
Mob,?, phase:
vacuo.
- mobile phase A: dissolve 3.52 g of potassium dihydrogen
phosphate R and 7.26 g of disodium hydrogen phosphore Bacterial endotoxins (2.6.14)
dihydrate R in 1000 mL of water for chromaJography R; Less than 0.14 TO per European Pharmacopoeia Unit of
filter and degas; aprotinln, if intended for use in the manufacture of
- mobile phase B: dissolve 3.52 g of potassium dihydrogen parenteral preparations without a further appropriate
phosphate R, 7.26 g of disodium hydrogen phosphare procedure for the removal of bacterial endoroxins.
dihydrate R and 66.07 g of ammonium sulfiue R in ASSAY
1000 mL of waterfor chromatography R; fiJter and degas; The activity of aprotinin is determined by measuring its
inhibitory action on a solution of trypsin of known activity.
Tim. M.obile pha!e A M.obile phase B The inhibiting activity of the aprotinin is calculated from the
(min) (per cent VIP) (per cent VIJ')
difference between the initial activity and the residual- activity
o ~ 21 92 ---> 64 8 --> 36
of the trypsin.
21 - 30 64 ---> 0 36 ---> 100
The inhibiting activity of aprotinin is expressed in European
Pharmacopoeia Unirs. I Ph. Eur. U. inhibits 50 per cent of
Flow rate 1.0 mUmin. the enzymatic activity of 2 microkatals of trypsin.
Delation Spectrophotometer at 210 run.
Injection 40 ur,
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Use a reaction vessel with a capacity of about 30 ml., IMPURITIES

provided with:
- a device that will maintain a temperature of 25 ± 0.1 °c; H - Arg-Pro- Asp-Phe - Cys - Leu-Glu- Pro-Pro - Tyr-
o •
- a stirring device, such as a magnetic stirrer; ~ ~ ~~~-~-~-lle-~-~-
- a lid with 5 holes for accommodating the electrodes, me ~-P~-~-Asn~ ~ ~ ~ "
~-Cys
tip of a burette, a tube for me admission of nitrogen and
the introduction of the reagents.
An automatic or manual titration apparatus may be used.
Gin Thr Phe Val Tyr Gly Gly Cys Arg A1a-
~-~-~-Asn-~e-~-k-~ ~~
" .
In the latter case the burette is graduated in 0.05 mL and the Cys - Mel- Arg -Thr - eys - Gly- OH "
pH-meter is provided with a wide reading scale and glass-
silver-silver chloride or other suitable electrodes.
"
Test solution Prepare a solution of the substance to be A. aprotinin-(1-56)-peptide,
examined in 0.0015 M borate buffer ,oIution pH 8.0 R
H- Arg- Pro- Asp-Phe -eys- Leu-Glu-Pro- Pro- Tyr-
expected to contain 1.67 Ph. Eur. UJrnL (about 0.6 mg o •
(m mg) per millilitre). ~ ~ ~-Cys~-~-~-~-~-~-
Trypsin solution Prepare a solution of trypsin BRP containing Tyr-P~-Tyr-Asn Ala "
lys Ala Gfy Leu Cys-
about 0.8 microkatals per mdlilitre, using 0.001 M
hydrochlori< acid as the solvent. Use a freshly prepared
solution and keep in iced water.
Gh Thr Phe Val Tyr Gly Gty-Cys kg
~-~-~-Asn-~e-~-k-~
..
"A1a-
~-Asp
Trypsin and apronnin solu,ion To 4.0 rnL of the trypsin Cys-~I-~-~-Cys-~-~-OO "
solution add 1.0 mL of the test solution. Dilute inunediately "
to 40.0 rnLwith 0.0015 M borate buffer,oIu'ionpH 8.0 R.
Allow to stand at room temperature for 10 min and then B. aprotinin-(1-57)-peptide,
keep in iced water. Use within 6 h of preparation.
~-~-~-Asp-~-Cys-~-~-~-~-~-
DiJUle trypsin solution Dilute 0.5 mL of the trypsin solution o u
to 10.0 rnL with 0.0015M borate buffer solution pH 8.0 R. ~ ~ ~Cys~-~-~-I~-~-~-
Allow to stand at room temperature for 10 min and then ~-Phe-~-Asn~ ~ ~

n
Gty~u-Cys "
keep in iced water.
Gl:n Tbr -Phe Val Tyr Gly Gly Cys Arg A1a-
Maintain an atmosphere of nitrogen in the reaction flask and
stir continuously; introduce 9.0 rnL of 0.0015 M borate buffer
~-~-Asn-~-Phe-~-~-~ ~-~ "
,olution pH 8.0 Rand 1.0 rnL of a freshly prepared 6.9 yJL Cys-M~-~-fu-Cys-~-GIy-~-~ "
solution of ben;wylarginine ethylester hydrothloride R. Adjust to "
pH 8.0 with 0.1 M sodinm hydroxide. When the temperarure
has reached equilibrium at 25 ± 0.1 'C, add 1.0 rnL of the C. (5-oxoprolyl)aprotinin (pyrogiutamylaprotinin).
trypsin and aprotinin solution and stan a timer. Maintain at _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""E<I
pH 8.0 by the addition of 0.1 M sodium hydroxide and note
the volume added every 30 s. Continue the reaction for
6 min. Determine the number of millilitres of 0.1 M sodium
hydroxide used per second (nl roL). Carry out, under the Aprotinin Concentrated Solution
same conditions, a titration using 1.0 mL of the dilute
trypsin solution. Determine the number of millilitres of (Ph. Bur. monograph 0579)
0.1 M sodium hydroxide used per second (n2 rnL).
H-Arg-Pro-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr-
Calculate the aprotinin activity in European Pharmacopoeia
Units per milligram using the following expression: ~-Gty-~-Cys-~-~-~-I~-~-~-
•
~-P~-~-~~ ~ ~ ~ "
~-Cys
Gin Thr Pile Val Tyr Gly Gly-Cys-Arg
Lys-Arg-Asn-Asn-P~-Lys-ser-A1a
.
"A1a-
Glu Asp-
The estimated activity is not less than 90 per cent and not
more than 110 per cent of the activity stated on the label.
Cys -Mel-Arg- Thr- Cys -Gly -Gly - Afa- OH "
"
STORAGE
.In an airtight) tamper-evident container, protected from light. 6511
LABELLING Action and use

The label ,rates: Antifibrinolytic.
- the number of European Pharmacopoeia Units of
""E<I ~ _
aprotinin activity per milligram;
- where applicable, that the substance is suitable for use in DEFINITION
the manufacture of parenteral preparations. Aprotinin concentrated solution is a solution of aprotinin, a
polypeptide consisting of a chain of 58 amino acids, which
inhibits stoichiometrically the activity of several proteolytic
enzymes such as chymotrypsin, kallikrein, plasmin and
trypsin. It contains not less than 15.0 Ph. Eur. U.
of aprotinin activity per millilitre.
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2022 Aprotinin 1-195
PRODUCTION Des-Ala-aprotinin and des-Ala-des-Gly-aprotinin

The animals from which aprotinin is derived must-fulfil the Capillary zone electrophoresis (2.2.47): use the normalisation
requirements for the health of animals suitable forhuman procedure.
consumption. Test solution Dilute the preparation to be examined in
The method of manufacture is validated to demonstrate that water R to obtain a concentration of not less than
the product, if tested, would complywith the following test. I Ph. Eur. U.lmL
Histamine (2.6. Iff) Reference solution Dilute aprotinin solution BRP in water R to
Maximum 0.2 ~g of histamine base per 3 Ph. Eur. U. obtain the same concentration as the testsolution.
Capillary:
CHARACTERS - material: uncoated fused silica;
Appearance - size: effective length = 45-60 cm, Q) = 75 um,
Clear, colourless liquid. Temperature 25 ·C.
IDENTIFICATION CZE buffer Dissolve 8.21 g of potassium dihydrogen
A. Thin-layer chromatography (2.2.27). phosphate R in 400 mL of wa"r R, adjust to pH 3.0 with
Testsolution Solution S (see Tests). phosphon·c acid R, dilute to 500.0 mL with water R and filter
Reference solution Dilute aprotinin solution BRP in waler R to through a membrane filter (nominal pore size 0.45 IJm).
obtain a concentration of 15 Ph. Bur. UJmL. Detection Spectrophotometer at 214 nrn.
Piau TLC "Jim gel G pkue R. Between-ron "·tlsing Rinse the capillary for at least I min
with 0.1 M sodium hydroxide filtered through a membrane
Mabile phase water R, glacial acetic acid R (80:100 VIV)
filter (nominal pore size 0.45 pm) and for 2 min with the
containing 100 gIL of sodium acetate R.
CZE buffer.
Appliauion 10 ~. Injection Under pressure or vacuum (forexample, 3 s at a
Development Overa path of 12 em. differential pressure of 3.5 kPa).
Drying In air. Migration Apply a field strength of 0.2 kYlcm, using the
Deteaion Spray with a solution of 0.1 g of ninhydrin R in a CZE buffer as the electrolyte in both buffer reservoirs.
mixture of 6 mL of a 10 gIL solution of cupric chloride R, Run time 30 min.
21 mL of glacial acetic acid Rand 70 mL of anhydrous ldentificau·on of impurities Use the electropherograrn supplied
ethanol R. Dry the plate at 60 "C. with aprotinin solucion BRP and the electropherogram
ResuJu The principal spot in the chromatogram obtained obtained with the reference solution to identify the peaks due
with me testsolutionis similar in position, colourand size to to impurities A and B.
the principal spot in the chromatogram obtainedwith the Relativemigration With reference to aprotinin (migration
reference solution. =
time about 22 min): impurity A =ahout 0.98;
B. Determine the ability of the preparation to be examined to =
impurity B about 0.99.
inhibit trypsin activity using the method described below. System suitability Reference solution after at least 6
Testsolution Dilute 1 mL of solution S to 50 mL with bufftr injections:
solution pH 7.2 R. - migration time: aprotinin = 19.0 min to 25.0 min;
Trypsin solution Dissolve 10 mg of trypsin BRP in 0.002 M - resolution: minimwn 0.8 between the peaks due to
hydrochlori< acid and dilute to 100 mL with the same acid. impurities A and B; minimum 0.5 between the peaks due
to impurity Band aprotinin;
Casein so/ucion Dissolve 0.2 g of casein R in buffer solution
- peak distllbution: the electropherogram obtained is
pH 7.2 R and dilute to 100 mL with the same huffer
qualitatively and quantitatively similar to the
solution.
electropherogram supplied with aprotinin solution BRP;
Precipitating solution glacial acetic acid R, water R, anhydrous - heightof the principal peak: at least 1000 times the height
ethanol R (1:49:50 VIVIV). of the baseline noise. If necessary, adjust the sampleload
Mix I mL of the test solution with 1 mL of the trypsin to give peaksof a sufficient height
solution. Allow to stand for 10 min and add 1 mL of the Limits:
casein solution. Incubate at 35 DC for 30 min. Cool In iced - impun'ty A: maximum 8.0 per cent;
water and add 0.5 mL of me precipitating solution. Shake - impun'ty B: maximum 7.5 per cent.
and aUow to standat room temperature for 15 min.
Pyroglutarnyl-aprotinin and related compounds
The solution is cloudy. Carry out a blank test under the
same conditions using bufftrsolution pH 7.2 R instead of the
procedure.
test solution. The solution is not cloudy.
Test solution Dilute the preparation to be examined in
TESTS mobilephase A to a concentration of about
Solution S 5 Ph. Eur. U.lmL.
Prepare a solution containing 15 Ph. Eur. U.lmL, if
Reference solution Dissolve the contents of a vial of aprotinin
necessary by dilution, on the basisof the activity stated on
for system suitability CRS in 2.0 mL of mobile phase A.
the label.
Column:
Appearance of solution - size: l = 0.075 m, 0 = 7.5 rnm;
Solution S is clear (2.2.1). - stationary phase: strong cation-exchange silica gelfor
Absorhance (2.2.25) chromatography R (10 urn);
Maximum 0.80 by measuring at the absorption maximum at - temperature: 40 "C.
277 run. Mobile phase:
Prepare a solutioncontaining 3.0 Ph. Eur. U./rnL. - mobile phase A: dissolve 3.52 g of potassium dihydrogen
phospho" Rand 7.26 g of disadium hydrogen phospho"
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dihydrate R in 1000 mL of waterfor chromawgraphy R; Evaporate 25.0 mL to dryness in a water-bath, dry the
filter and degas; residue at 110°C for 15 h and weigh. From the mass of the
- mobile phase B: dissolve 3.52 g of potassium dihydrogen residue and the activity determined as described below,
phosphate R, 7.26 g of disodium hydrogen pho,phate calculate the number of European Pharmacopoeia Units per
dihydrate Rand 66.07 g of ammonium su!fate R in milligram of dry residue.
1000 mL of waterfor chromawgraphy R; filter and degas; Baclerial endotoxin. (2.6.14)
Less than 0.14 ill per European Pharmacopoeia Unit of
Tim. Mobile phase A Mobile phase B aprotinin, if intended for use in the manufacture of
(min) (per cent VIl? (per cent V/J?
parenteral preparations without a further appropriate
o ~ 21 92 -+ 64 8 36
64 _ 0
21 - 30 36 100
ASSAY
The activity of aprotinin is determined by measuring its
FkJw rate 1.0 rnUmin.
inhibitory action on a solution of trypsin of known activity.
Detection Spectrophotometer at 210 ron. The inhibiting activity of the aprotinin is calculated from the
Injection 40 ~L. difference between the initial activity and the residual activity
Relative retention With reference (Q aprotinin (retention of the trypsin.
time = =
17.0 min 10 20.0 min): impurity C about 0.9. The inhibiting activity of aprotinin is expressed in European
System suitability Reference solution: Pharmacopoeia Units. 1 Ph. Eur. U. inhibits 50 per cent of
- resolution: minimum 1.5 between me peaks due to the enzymatic activity of 2 rnicrokatals of trypsin,
impurity C and eprorinin; Use a reaction vessel with a capacity of about 30 mL,
- symmetry faaor: maximum 1.3 for the peak due to provided with:
aprotinin. - a device that will maintain a temperature of 25 ± 0.1 "C,
limits: - a stirring device, such as a magnetic stirrer;
- impun",y C: maximwn 1.0 per cent; - a lid with 5 holes for accommodating the electrodes, the
- any other impun"ty: maximum 0.5 per cent; tip of a burette, a tube for the admission of nitrogen and
- sum of impurities otherthan C: maximum 1.0 per cent. the introduction of the reagents.
Aprodnin ollgomers An automatic or manual titration apparatus may be used.
Size-exclusion chromatography (2.2.30): use the In the latter case the burette is graduated in 0.05 mL and the
normalisation procedure. pH-meter is provided with a wide reading scale and glass-
Test so/utien Dilute the preparation to be examined in silver-silver chloride or other suitable electrodes.
water R to obtain a concentration of about 5 Ph. Bur. UJmL. Test ,oIution With 0.0015 M borate buffer 'olutionpH 8.0 R
Reference solution Treat the substance to be examined to prepare an appropriate dilution (D) of the aprotinin
obtain about 2 per cent aprotinin oligomers. For example, concentrated solution expected, on me basis of the stated
heat freeze-dried aprotinin at about 110°C for about 4 h. porency, to contain 1.67 Ph. Eur. U.lmL.
Then dissolve in water R to obtain a concentration of about Trypsin solution Prepare a solution of trypsin BRP containing
5 Ph. Bur. U.lmL. about 0.8 rnicrokatals per millilitre, using 0.001 M
Column 3 columns coupled in series: hydrochloric acid as the solvent. Use a freshly prepared
- size: I ::;; 0.30 m, 0 ::;; 7.8 mID; solution and keep in iced water.
- stationary phase: hydrophilic silica gelfor chromawgraplty R Trypsin and aprotinin solution To 4.0 mL of the trypsin
of a grade suitable for fractionation of globular proteins in solution add 1.0 mL of the test solution. Dilute immediately
the relative molecular mass range of 20 000 to 10 40.0 mL with 0.0015 M borate /niffer 'olutionpH 8.0 R.
10000000 (8 um). Allow to stand at room temperature for 10 min and then
Mobile phase acetonitrile R, glacialacetic acid R. waterfor keep in ked water. Use within 6 h of preparation.
chromawgraplty R (2:2:6 VIVIV); filter and degas. Dilute trypsin solution Dilute 0.5 mL of the trypsin solution
Flow rate 1.0 rnUmin. to 10.0 mL with 0.0015 M borate buffer ,oIution pH 8.0 R.
Allow to stand at room temperature for 10 min and then
keep in ked water.
Injection 100 ~L. Maintain an atmosphere of nitrogen in the reaction flask and
Run time 40 min. stir continuously; introduce 9.0 mL of 0.0015 M borate buffer
Relative retention With reference to aprotinin monomer ,oIut;"n pH 8.0 Rand 1.0 mL of a freshly prepared 6.9 gIL
(retention time = 24.5 min to 25.5 min): aprotinin solution of benzoylarginine ethylester hydrochloride R. Adjust to
dimer e about 0.9. pH 8.0 with O. I M 'odium hydroxide. When the temperature
Systemsuitability Reference solution: has reached equilibrium at 25 ± 0.1 'C, add 1.0 mL of the
- resolution: minimwn 1.3 between the peaks due to trypsin and aprotinin solution and start a timer. Maintain at
aprotinin dimer and monomer; pH 8.0 by the addition of 0.1 M 'odium hydroxide and note
- symmetry factor: maximum 2.5 for the peak due to the volume added every 30 s. Continue the reaction for
aprotinin monomer. 6 min. Determine the number of millilitres of 0.1 M sodium
Limit: hydroxitk used pel second (n, mL). Carry oUI, under the
- totai: maximum 1.0 per cent. same conditions, a titration using 1.0 mL of the dilute
trypsin solution. Determine the number of millilitres of
Specific activity of the dry residue 0.1 M sodium hydroxide used per second {n, rnI4.
Minimum 3.0 Ph. Bur. U. of aprotinin activity per milligram
Calculate the aprotinin activity in European Pharmacopoeia
of dry residue.
Units per millilitre using the following expression:
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2022 Arachis Oil 1-197
4000(2n, - n,) x D
CHARACTERS
D dilution factor of the aprotinin concentrated solurion to be Appearance
examined in order [0 obtain a solution containing Clear, yellowish) viscous liquid.
1.67 Ph. Eur. UJrnL
Solubility
Very slightly soluble in ethanol (96 per cent), miscible with
The estimated activity is not less than 90 per cent and not
light petroleum.
more chan 110 per cent of the activity stated on the label.
Relative density
STORAGE About 0.915.
In an airtight, tamper-evident container, protected from light.
It solidifies at about 2 "C.
LABELLING
IDENTIFICATION
The label states:
- the number of European Phannacopoeia Units of
aprotinin activity per millilitre; Second identification: A.
- where applicable, that the substance is suitable for use in A. Identification of fatty oils by thin-layer chromatography
the manufacture of parenteral preparations. (2.3.2).
IMPURITIES Results The chromatogram obtained is similar to the
H-Arg-Pro-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr- B. Composition of fat{y acids (see Tests).
, rc
fu ~ ~cp~-~-~-~-~-~- TESTS
~-P~-~-Asn~ ~ ~ ~~-Cys " Acid value (2.5.1)
GIn Thr Phe-Val Tyr Gly GIy-Cys kg A1a- " Maximum 0.5, determined on 10.0 g.
Peroxide value (2.5.5, MethodA)
~-~-~-Asn-P~-~-k-~ ~-Asp" Maximum 5.0.
Cys-Met - Arg- Thr - Cys - GIy- OH " UnsaponlfiabIe matter (2.5.7)
" Maximum 1.0 per cent, determined on 5.0 g.
A. aprotinin-(1-56)-peptide, A1kallne Impurities (2.4.19)
It complies with the test.
H-Arg-Pro-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr-
,
Thr-~ ~-~s~-~-Arg-~-~-Arg-
'" Composition offatty acids (2.4.22, Mahod A)
Tyr-Phe-Tyr-Asn Ala Lys Ala Gly Leu "
Cys- Composition of thefatty-acidfraction of the oil:
Gin Thr - Phe Val Tyr Gly GIy- Cys - Arg A1a-" - saturated fO/1Jl acids of chain length less than C,,: maximum
0.4 per cent;
Lys-Arg-Asn-Asn-Phe-Lys-Ser-Ala Glu "
Asp-
- palmiticacid: 5.0 per cent to 14.0 per cent;
Cys - Met - Arg - Thr - Cys - Gly - Gly - OH " - stearic acid: 1.3 per cent to 6.5 per cent;
" - oleic acid: 35.0 per cent to 76.0 per cent;
- linoleic add: 8.0 per cent to 43.0 per cent;
B. aprotinin-(l-57)-peptide, - linolenic acid: maximum 0.6 per cent;
- arachidic add: 0.5 per cent to 3.0 per cent;
~-~-~-Asp-Phe-~-~-G~-~-~-~-
- eicosenoic acid: 0.5 per cent to 3.0 per cent;
, ~ ~ ~-Cys ~-~-Arg-~-~-Arg-
"
- behenic acid: 1.0 per cent to 5.0 per cent;
~-P~-~-~n~ ~ ~ ~~-Cys " - erucic acid: maximum 0.5 per cent;
- lignocetic add: 0.5 per cent to 3.0 per cent.
Gin Thr-Phe Val Tyr Gly Gly Cys Arg Ala- " Water (2.5.32)
~-Arg-~-~n-Phe-~-~-~ ~-Asp" Maximum 0.1 per cent, determined on 1.00 g.
n
C~-M~-~-~-Cys-~-GIy-~-OO
STORAGE
" In a well-filled container, protected from light.
C. (5-oxoprolyl)aprotinin (pyroglutamylaprotinin). -------- PIIE"
_____________________ "'E"
Hydrogenated Arachis Oil

Arachis Oil Hydrogenated Peanut Oil
Peanut Oil (ph. Bur. monograph 1171)
(Refined Arachis Oil, Ph. Bur. monograph 0263) "'E" ---
Preparation DEFINITION
Arachis Oil Enema Oil obtained by refining, bleaching, hydrogenating and
"'E" _ deodorising oil obtained from the shelled seeds of Arachis
hypogaea L. Each type of hydrogenated arachis oil is
DEFINITION characterised by irs nominal drop point.
The refined fatty oil obtained from the shelled seeds of
Arachis hypogaea L. A suitable antioxidant may be added.
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1-198 Arginine 2022
CHARACTERS STORAGE
Appearance , Protected from light.
White or faintly yellowish, soft mass which melts to a clear,
LABELLING
pale yellow liquid when heated.
The label states the nominal drop point
Solubility _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "'81
Practicallyinsoluble in water, freely soluble in methylene
chloride and in light petroleum (bp: 65-70 "C), very slightly
IDENTIFICATION Arginine
Secondidentification: A, B. (ph. Eur. monograph 0806)
A. Drop point (see Tests).
H H iNti:!
B. Identification offatty oils by thin-layer chromatography H,N
(2.3.2). Y N ~co,H
V
Results The chromatogram obtained is similar to the NH

C. Composition or fatty acids (see Tests). 174.2 74-79-3
TESTS Action and use
Drop point (2.2.17) Amino acid; nutrient.
32 "C to 43 "C, and within 3 "C of the nominal value.
Pl>EII ~_
Acid value (2.5.1)

Maximwn 0.5. DEFINITION
Dissolve 10.0 g in 50 mL of the prescribed solvent by (2S)-2-Amino-5-guanidinopentanoic acid.
heating on a water-bath. Product of fermentation or of protein hydrolysis.
Peroxide value (2.5.5, Method A) Content
Maximwn 5.0. 98.5 per cent to 101.0 per cent (dried substance).
Dissolve 5.0 g in 30 mL of the prescribed solvent by heating CHARACTERS
on a water-bath. Appearance
Unsaponifiable matter (2.5.7) White or almost white, crystalline powder or colourless
Maximum 1.0 per cent, determined on 5.0 g. crystals) hygroscopic.
Alkaline impurities (2.4.19) Solubility
It complies with the test. Freely soluble in water, very slightly soluble in ethanol
Composition of fatty acids (96 per cent).
Gas chromatography (2.4.22, Method A) with the following IDENflFICATION
modifications. Use the mixture of calibrating substances in First identification: A.J C.
Table 2.4.22.-3. Second identification: A.J B.J D, E.
Column: A. Specific optical rotation (see Tests).
B. Solution S (see Tests) is strongly alkaline (2.2.4).
- size: I :::; 25 m, 0 :::; 0.25 mm;
- statIOnary phase: cyanopropy/po/ysiloxane R (film thickness C. Infrared absorption spectrophotometry (2.2.24).
0.2 pm). Comparison arginine CRS.
Gooiergas helium for chromatography R. If the spectra obtained show differences, dry the substance to
Flow rate 0.7 mUmin. be examined and the reference substance in an oven at
105 °C and record new spectra.
Splirralro 1:100.
D. Thin-layer chromatography (2.2.27).
Temperature:
- column: 180 °C for 20 min; Testsolution Dissolve 10 mg of the substance to be
- injection port and detector: 250 °C. examined in a 10.3 gIL solution of hydrochlori< acid Rand
dilute to 50 mL with me same solution.
Reference solution Dissolve 10 mg of arginine CRS in a
Composition of thefauy-add fraaion of the substance:
10.3 gIL solution of hydrochlori< add R and dilute to 50 mL
- saturated fouy acids of chain length less than c,.,: maximum
with the same solution.
0.5 per cent;
- myristic acid: maximum 0.5 per cent; Plate TLC silica gelplateR.
- palmitic acid: 5.0 per cent to 16.0 per cent; lvlobr7e phase concentrated ammonia R, 2-propanol R
- stearic acid: 3.0 per cent to 19.0 per cent; (30:70 VIV).
- oleic acidand isomers: 54.0 per cent to 78.0 per cent; Applica'ron 5 ~L.
- linoleic acid and isomers: maximum 10.0 per cent; Development Over 2/3 of the plate.
- arachidic add: 1.0 per cent to 3.0 per cent;
- eicosenoic acids: maximum 2.1 per cent; Drying At 105 "C until the armnonia disappears completely.
- behenic acid: 1.0 per cent to 5.0 per cent; Detection Spray with ninhydrin solution R and heat at 105 °C
- erucic acidand isomers: maximum 0.5 per cent; for 15 min.
- lignoceric acid: 0.5 per cent to 3.0 per cent
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2022 Arginine 1-199
Results The principal spot in the chromatogram obtained Limits:

with the test solution is similar in position, colour and size [Q - any ninhydrin-positive substance: for each impurity,
the principal spot in the chromatogram obtained with the maximum 0.2 per cent;
reference solution. - total: maximum 0.5 per cent;
E. Dissolve about 25 mg in 2 mL of waterR. Add I mL of - reporting threshold: 0.05 per cent.
rJ.~naphthoi solution Rand 2 mL of a mixture of equal volumes The thresholds indicated under Related substances
of strong sodium hypochlm;ce solution R and waterR. A red (Table 2034.-1) in the general monograph Substances for
colourdevelops. phamlaceutical use (2034) do not apply.
TESTS Chlorides (2.4. if)
Solution S Maximum 200 ppm.
Dissolve 2.5 g in distilled water R and dilute to 50 mL with To 5 mL of solution S add 0.5 mL of dilute niuic acid Rand
the same solvent. dilute to 15 mL with water R.
Appearance of solution Sulfates (2.4.13)
Solution S is clear (2.2.1) and not more intensely coloured Maximum 300 ppm.
than reference solution BY6 (2.2.2, Method11). To IO mL of solution S, add 1.1 mL of dilute hydrochlori<
Specific optical rotation (2.2.7) acid R and dilute to 15 mL with distilled water R.
+ 25.5 to + 28.5 (dried substance). Ammonium
Dissolve 2.00 g in hydrochlori< acidRl and dilute to 25.0 mL Amino acid analysis (2.2.56) as described in the test for
with the same acid. ninhydrin-positive substances with the following
Ninhydrin-positive substances modifications.
Amino acid analysis (2.2.56). For analysis, use Method 1. Injection Test solution, reference solution (c) and blank
The concentrations of the test solution and the reference solution.
solutions may be adapted according to the sensitivityof the Limit:
equipment used. The concentrations of all solutions are - ammonium at 570 nm: not more than the area of the
adjusted so that the system suitability requirements described corresponding peak in me chromatogram obtained with
in general chapter1.1.46 are fulfilled, keeping the ratios of reference solution (c) (0.02 per cent), taking into account
concentrations between all solutions as described. the peak due to ammonium in the chromatogram
Solution A water R or a sample preparation buffer suitable obtained with the blanksolution.
for the apparatus used. . Iron (2.4.9)
Test solution Dissolve 30.0 mg of the substance to be Maximum 10 ppm.
examined in solution A and dilute to 50.0 mL with In a separating funnel, dissolve 1.0 g in 10 mL of dilute
solution A. hydrochlori< acid R. Shake with 3 quantities, each of 10 mL,
Reference solution (a) Dilute 1.0 mL of the test solution to of methyl isobutyl ketone Rt, shakingfor 3 min each time.
100.0 mL with solution A. Dilute 2.0 mL of this solution to To the combined organic layers add IO mL of water Rand
10.0 mL with solution A. shake for 3 min. Use me aqueous layer.
Reference ,oIUlion (b) Dissolve 30.0 mg of proline R in Loss on drying (2.2.32)
solution A and dilute to 100.0 mL with solution A. Dilute Maximum 0.5 per cent, determined on 1.000 g by drying in
1.0 mL of the solution to 250.0 mL with solution A. an oven at 105°C.
Reference solution (c) Dilute 6.0 mL of ammonium standard Sulfated ash (2.4.1if)
solution (100 ppm NHJ R to 50.0 mL with solution A. Dilute Maximum 0.1 per cent, determined on 1.0 g.
1.0 mL of this solution to 100.0 mL with solution A.
ASSAY
Reference solution (d) Dissolve 30 mg of isoleucine Rand Dissolve0.150 g in 50 mL of water R. Titrate with 0.1 M
30 mg of leucine R in solution A and dilute to 50.0 mL with hydrochltJrU acid, determining the end-point potentiometricaUy
solution A. Dilute 1.0 mL of the solution to 200.0 mL with (2.2.2ff).
solution A.
I mL of 0.1 M hydrochlori< acid is equivalent to 11.42 mg of
Blank solution Solution A. CoH14N,02·
Inject suitable, equal amounts of the test, blank and reference
STORAGE
solutions into the amino acid analyser. Run a program
suitable for the determination of physiological amino acids.
System suiuzbility Referencesolution (d): IMPURITIES
- resolution: minimum 1.5 between the peaks due to Otherdetectable impurities (thefollowing substances wauld, if
isoleucine and leucine. present at a sufficient/eve!, be detected by one or other of tire tests
CalculatWn of percentage contents: in the monograph. They arelimited by thegeneral aaeplarn:e
- for any ninhydrin-positive substancedetected at 570 nm, criterion for other/unspecified impurities. It is therefore not
use the concentration of arginine in reference solution (a); neussary to identify these impun'ties for demonstration of
- for any ninhydrin-positive substance detected at 440 om, compliance. See also 5.10. Control a/impurities in substances/or
use the concentration of proline in reference solution (b); pharmaceutical use) A, B, C.
if a peak is above the reporting threshold at both
wavelengths, use the result obtained at 570 om for
quantification.
A. (2S)-2,6-diaminohexanoic acid (lysine),
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1-200 Arginine Aspartate 2022
Dissolve 2.50 g in dilute hydrochloric acidR and dilute to

25.0 mL with the same acid.
B. (2S)-2-arnino-5-(carbamoylamino)pentanoic acid Test solution (a) Dissolve 0.20 g of the substance to be
(citrulline), examined in water R and dilute to 10 mL with me same
solvent.
H NH,
Test solution (b) Dilute I rnL of test solution (a) to 10 mL
H,N~ CO,H with water R.
Reference solution (a) Dissolve 25 mg of arginine Rand
C. (2S)-2,5-diaminopentanoic acid (ornithine). 25 mg of aspartic acid R in water R and dilute to 25 rnL with
_____ ~ ----' PhE" the same solvent.
Reference solution (b) Dilute 2 mL of reference solution (a)
to 50 rnL with water R.
Ptate TLC silIca gel G plate R.
Arginine Aspartate ***
*** *** Mobile phase ammonia R, propanol R (36:64 VIV).
Application 5 ~L.
(Ph. Bur. monograph 2096) ***
H H .•N~ Drying At 100-105 °C for 10 min.
H,NyN~Co,H Deuaion Spray with ninhydn"n solution R and heat at
NH 100-105 °C for 10 min.
307.3 7675-113-4 - the chromatogram shows 2 clearly separated principal
spots.
Action and use Limit Test solution (a):
Amino acid; nutrient. - any impun"ty: any spots, apart from the 2 principal spots,
PhE" _ are not more intense than each of the 2 principal sPO[S in
DEFINITION (0.2 per cent).
(2S)-2-Amino-5-guanidinopentanoic acid (2S)-2- Chlorides (2.4.4)
aminobutanedioate. Maximum 200 ppm.
Content Dilute 2.5 rnL of solution S to 15 rnL with water R.
Sulfutes (2.4.13)
CHARACTERS Maximum 300 ppm.
Appearance To 0.5 g add 2.5 mL of dilute hydrochloric acid R and dilute
White or almost white granules or powder. to 15 rnL with distilled water R. Examine after 30 min.
Solubility Ammonium (2.4.1)
Verysoluble in water, practically insoluble in alcohol and in Maximwn 100 ppm, determined on 100 mg.
methylene chloride.
IDENTIFlCATION Maximum 0.5 per cent, determined on 1.000 g by drying in
A. Specific optical rotation (see Tests). an oven at 60 °C for 24 h.
B. Infrared absorption spectrophotometry (2.2.24). Sulfated ash (2.4.14)
Comparison arginine aspartate CRS. Maximum 0.1 per cent, determined on 1.0 g.
C. Examine the chromatograms obtained in the test for ASSAY
ninhydrin-positive substances. Dissolve 80.0 mg in 2 rnL of anhydrous formic acid R.
Results The 2 principal spots in the chromatogram obtained Add 50 rnL of anhydrous acetic acid R. Titrate with 0.1 M
with test solution (b) are similar in position) colour and size perchlonc acid, determining the end-pointpotentiometrically
to the 2 principal spots in the chromatogram obtained with (2.2.21J).
reference solution (a). 1 rnL of 0.1 M perchlotic acid is equivalent to 10.24 mg
TESTS of C,oH21N,06'
Solution S __ ~ PhE"
Dissolve5.0 g in carbon dioxide-free water R and dilute (0
than reference solution Y, (2.2.2, Method lI).
pH (2.2.3)
+ 25 to + 27 (dried substance).
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2022 Arginine Hydrochloride 1-201
Arginine Hydrochloride *** TESTS

** ** Solution S
**** * Dissolve 2.5 g in distilled water R and dilute to 50 mL with
the same solvent.
H H ,NH2 Appearance of solution
H,N'V'N~
II C0
• Hel Solution S is clear (2.2.1) and not more intensely coloured
2H
NH than reference solution BY6 (2.2.2.1 JWethod 1/).
210.7 //19-34-2 + 21.0 to + 23.5 (dried substance).
Dissolve 2.00 g in hydrochloric acid Rl and dilute to 25.0 mL
Action and use with the same acid.
Amino acid; nutrient.
Preparations Amino acid analysis (2.2.56). For analysis) use Method 1.
Arginine Hydrochloride Infusion
The concentrations of the test solution and the reference
Arginine Hydrochloride Oral Solution solutions may be adapted according to the sensitivity of the
Arginine Hydrochloride Sterile Concentrate equipment used. The concentrations of all solutions are
!'hE<r _
adjusted so that the systemsuitability requirements described
in general chapter 2.2.46 are fulfilled) keeping the ratios of
DEFINlTION concentrations between all solutionsas described.
(2S)-2-Amino-5-guanidinopentanoic acid hydrochloride. Solution A waler R or a samplepreparation buffer suitable
Product of fermentation or of protein hydrolysis. for the apparatus used.
Content Test solution Dissolve 30.0 mg of the substance to be
98.5 per cent to 101.0 per cent (dried substance). examined in solutionA and dilute to 50.0 mL with
solution A.
CHARACTERS
Reference solution (a) Dilute 1.0 mL of the test solution CO
Appearance
100.0 mL with solutionA. Dilute 2.0 mL of this solution to
crystals.
Reference solution (b) Dissolve 30.0 mg of proline R in
Solubility
solution A and dilute to 100.0 mL with solution A. Dilute
Freely soluble in water, very slightly soluble in ethanol 1.0 mL of the solutionto 250.0 mL with solutionA.
(96 per cent).
Reference solution (c) Dilute 6.0 mL of ammonium standard
IDENTIFICATION solution (/00 ppm NH.J R to 50.0 mL with solution A. Dilute
First idenufication: A, B, E. 1.0 mL of this solution to 100.0 mL with solution A.
Second identification: A, C, D, E. Reference solution (d) Dissolve 30 mg of isoleucine Rand
A. Specific optical rotation (see Tests). 30 mg of leucine R in solution A and dilute to 50 mL with
B. Infrared absorption spectrophotometry (2.2.24). solution A. Dilute I mL of the solution to 200 mL with
solution A.
Comparison arginine hydrochloride CRS.
Blank solution SolutionA.
Injectsuitable) equalamounts of the test) blank and reference
examined in water R and dilute to 50 mL with the same
suitable for the determination of physiological amino acids.
solvent.
Reference solution Dissolve 10 mg of arginine
hydrochloride CRS in water R and dilute to 50 mL with the
isoleucine and leucine.
same solvent.
Plate TLC silica gel plate R.
- for any ninhydrin-positive substance detected at 570 run)
lYlobile phase concentrated ammonia R, 2-propanol R use the concentration of arginine hydrochloride in
(30:70 VIV). reference solution (a);
Application 5 ~L. - for any ninhydrin-positive substance detected at 440 nm,
Development Over 2/3 of the plate. use the concentration of proline in reference solution (b);
Drying At 105°C until me ammonia disappears completely. if a peakis above the reporting threshold at both
wavelengths, use the resultobtained at 570 run for
Detection Spray with ninhydrin solution R and heat at 105 "C
quantification.
for 15 min.
Limits:
- atry ninhydn'n-positive substance: for each impurity)
with the test solutionis similar in position, colour and size to
maximum 0.2 per cent;
the principal spot in me chromatogram obtainedwith the
reference solution.
D. Dissolve about 25 mg in 2 mL of waterR. Add I mL of
The thresholds indicated underRelated substances
a-naphthol solution Rand 2 mL of a mixture of equal volwnes
of strong sodium hypochlorite solution R and water R. A red
colour develops.
E. It gives reaction (a) of chlorides (2.3.1).
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1-202 Argon 2022
Sulfates (2.4.13)
Argon ***
Maximum 300 ppm. *** ***
Dilute 10 mL of solution S to 15 mL with distilled waterR. (Ph. Bur. monograph 2407) ***
Ammonium Ar 39.95 744()-37-1
Amino acid analysis (2.2.56) as described in the test for PhE" _
modifications. DEFINITION
Injection Test solution, reference solution (c) and blank Gas obtained by fractional distillation of ambient air.
solution. Content
Limit: Minimum 99.995 per cent VIV of Ar, calculated by
- ammonium at 570 nm: not more than the area of the deductionof the sum of impurities found when performing
corresponding peak in the chromatogram obtained with the test for impurities and me water content.
reference solution (c) (0.02 per cent), taking into account This monograph applies £0 argon for medicinal use.
the peak due to ammonium in me chromatogram
CHARACTERS
obtained with the blanksolution.
Appearance
Iron (2.4.9) Colourless gas.
Maximum 10 ppm.
Solubility
In a separating funnel, dissolve 1.0 g in 10 mL of dilute
At 20°C and at a pressure of 101 kPa, 1 volume dissolves in
hydrochlotic acid R. Shake with 3 quantities, each of 10 ml., about 29 volumes of water.
of methyl isobutyl ketone Rl, shaking for 3 min each time.
To the combined organic layers add 10 mL of waterR and IDENTIFICATION
shakefor 3 min. Use the aqueous layer. A. Verify that the gas is not oxygen using a paramagnetic
analyser (2.5.27).
Los. on drying (2.2.32)
Maximum 0.5 per cent, determined on 1.000 g by drying in B. Gas chromatography (2.2.28).
an oven at 105 °C. Gas to be examined The substance to be examined.
Sulfated ash (2.4.14) Reference gas Use the following mixture of gases in
Maximum 0.1 per cent, determined on 1.0 g. argon Rl: methane Rl (5 ppm VIV), nitrogen Rl (5 ppm VIV),
oxygen R (5 ppm VIV).
ASSAY
Dissolve 0.180 gin 3 mL of anhydrousfonnic acidR. Column:
Add 30 mL of anhydrous acetic acid R. Titrate with 0.1 M - material: stainless steel)
perchloric acid, determining the end-pointpotentiometrically - size: 1= 2 m, 0 = 3 mm;
(2.2.21J). - statUmary phase: molecular sievefor chromatography R
(particle size 150-180 urn, pore size 0.5 nm).
I mL of 0.1 M perchwri< acid is equivalent to 21.07 mg of
Carrier gas helium for chroma",graphy R.
C.H , SCIN,02'
Flow rate 10 mllmin.
STORAGE
Temperature:
- column: 50 °c;
IMPURITIES - detee"'r. ISO 'C.
Otherdete<table impurities (thefollowing substances would, if Detection Thermal conductivity.
present at a sufficient level, be detected by cme or other of the tests
Jnje<tion 25 IlL.
criterion for other/unspecified impurities. It is therefore not System suitability Reference gas:
necessary to identify these impun"tits for demonstration of - resolulWn: minimwn 3.0 between the peaks due to
compliance. See also 5.10. Control of impurities in substances for argon/oxygen and nitrogen and minimum 2.0 between
pharmaceutical use) A, B, C. the peaksdue to nitrogen and methane.
with the gas to be examinedis similar in retention time to
reference gas.
A. (2S)-2,6-diaminohexanoic acid (lysine), TESTS
ImpuritIes
H NI<, Gas chromatography (2.2.28).
I<,N'V~~
II co,H Gas to be examined The substance to be examined.
o Reference gas Use the following mixture of gases in
argon Rl: methane Rl (5 ppm VIV), nitrogen Rl (5 ppm VIV),
B. (2S)-2-amino-5-(carbamoylamino)pentanoic acid oxygen R (5 ppm VIV).
(citrulline), Column:
H NI<,
=
- size: 1= 4 rn, 0 4 rom;
H~~Cl>.!H - stationary phase: moleadar sieve for chromawgraphy R
(particle size 150-180 urn, pore size 0.5 nm).
C. (2S)-2,5-diaminopentanoic acid (ornithine). Cartier gas argon Rl.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
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2022 Aripiprazole 1-203
Flow rate 70 mUmin. dihydroquinolin-2(lH)-one and 7-(4-iodobutoxy)-3,4-

Temperature: dihydroquinolin-2(lH)-one are genotoxic and are potential
- column: 80 °c; impurities in aripiprazole. These impurities are controlled by
- detector: 40 ·C. a suitable validated method.
Detection Discharge ionisation. CHARACTERS
Injection I mL. Appearance
Sample rale 100 mUmin. White or almost white crystals or crystalline powder.
Relative retention With reference to impurity C (retention Solubility
=
time = about 4.7 min): impurity A about 0.4; Practically insoluble in water, soluble in methylene chloride,
impurity B = about 0.7. very slightly soluble in ethanol (96 per cent).
System suitability Reference gas: It shows polymorphism (5.9).
- resolution: minimum 3.0 between the peaks due to IDENTIFICATION
impurities A and B and minimum 2.0 between the peaks Infrared absorption spectrophotometry (2.2.24).
due to impurities Band C.
Comparison an'piprazole CRS.
Limits: If the spectra obtained in the solid state show differences,
- impurily A: not more than the area of the corresponding
peak in the chromatogram obtained with the reference gas
substance separately in methylene chloride R J evaporate to
(5.0 ppm VflI);
- lotal: maximum 0.0040 per cent of the sum of me areas of
all the peaks (40.0 ppm VII'). TESTS
Water (2.5.28) Appearance of solution
Maximum 10.0 ppm VIV, determined using an electrolytic H intended for use in the manufacture of parenteral
hygrometer. preparations, the solution is clear (2.2.1) and not more
intensely coloured than reference solution GYj (2.2.2,
STORAGE Method11).
In gaseous or liquid slate, in suitable containers, complying Dissolve 0.5 g in a mixture of 10 volumes of acetic acid Rand
with the legal regulations. 90 volumes of anhydrous ethanol R and dilute to 20 mL with
IMPURITIES the same mixture of solvents. Sonicate for about 15 min,
Specified impuri,i., A, D. shaking occasionally, until dissolution is complete.
Other detectable impun"tiis B.. C. Related substances
A. oxygen, Liquid chromatography (2.1.29). Prosea the saluoons from
lighe
B. nitrogen,
C. methane,
Solwnr mixture acetiC acidR, methanol R, auron;m'/e R,
waw R (1:10:30:60 VIVIVII').
D. water.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEtr
Test solution Dissolve 50.0 mg of the substance £0 be
the solvent mixture. Dilute 5.0 mL of the solution £0
Aripiprazole 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
Reference solution (b) Dissolve 5 mg of the substance to be
examined and 5 mg of aripipmzo/e impuri/)' F CRS in the
rN~°Y'y~YO solvent mixture and dilute to 100 mL with the solvent
('yN~ VV mixture. Dilute 1 mL of the solution to 50 mL with the
solvent mixture,
yCI Reference solutWn (c) Dissolve 50.0 mg of aripiprazole CRS
in the solvent mixture and dilute to 50.0 mL with the solvent
CI
mixture. Dilute 5.0 mL of the solution to 50.0 mL with the
129722-12-9 solvent mixture.
C"H"C1,N,O, 448.4
Column:
Action and use - size: 1= 0.10 m, 0 = 4.6 mm;
Dopamine D 2 receptor antagonist; neuroleptic. - suuionasy phase: end-capped ocladecy/si/y/ silica gelfor
PhEtr _
ehromaUigraphy R (3 um}.
Mobile phase:
DEFINlTION - mobile phase A: aceronirrile R, 0.05 per cent VIV solution of
7-[4-[4-(2,3-DicWorophenyl)piperazin-I-yl)butoxy)-3,4- trifluoroacetic acid R (10:90 VII');
dihydroquinolin-2(1H)-one. - mobile phase B: 0.05 per cent VIV solution of tnfiuoroacetic
Content acid R, acetonitrile R (10:90 VII');
PRODUCTION
It is considered that impurities 7-(4-bromobutoxy)-3,4-
dihydroquinolin-2(1H)-one, 7-(4-cWorobutoxy)-3,4-
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1-204 Aripiprazole 2022
Tirn.e MoblJe phase A Moblle phase B

(min) (per cent VIP) (per cent V/I1
0-2 80 20
2 - 10 80 ---. 65 20 -i' 35
10 - 20 65 ..... 10 35 ...... 90
20·25 10 90
B. 1-(2,3-dicWorophenyl)pipera2ine,
InjecuOn 20 J.lL of me test solution and reference
Relative retention With reference to aripiprazole (retention
=
time about 11 min): impurity F about 1.1. =
System sU;UJbility Reference solution (b):
C. 7-[4- [4-(2-cWorophenyl) piperazin-I-yl] butoxy)- 3,4-
dihydroquinolin-ZfIffl-one,
aripiprazole and impurity F.
- for each impurity, use the concentration of aripiprazole in
Limits:
- unspecified impun"ties: for each impurity, maximum
0.10 per cent;
- total: maximwn 0.2 per cent;
- reporting threshold: 0.05 per cent. D. 7-[4- [4-(3-cWorophenyl)piperazin-I-yl)butoxy)-3,4-
Loss on drying (2.2.32) dihydroquinolin-2(1H)-one,
Dissolve 1.0 mg of the substance to be examined in 20 mL
of a 5.17 gIL solution of hydrochloric add R.
ASSAY E. 7-[4-[4-(2,3-dicWorophenyl)piperazin-l-yl)
Liquid chromatography (2.2.29) as described in the test for butoXYJquinolin-2(1H)-one,
Injeuion Test solution and reference solution (c). o H
System suitability Reference solution (c): r~~°Y'yNyO

- symmetry factor: maximum 2.0. ('yN,J VV
Calculate the percentage content of C23H27Cl2N302 taking
into account the assigned. content of an'piprazok CRS.
STORAGE
Protected from light. If the substance is sterile, store in a
'Y CI C1
F. 4-(2,3-dicWorophenyl)-I-[4-[(2-oxo-I,2,3,4-
sterile, airtight, tamper-evident container.
tetrahydroquinolin-7-yl)oxy]butyl)pipera2ine l-oxide,
LABELLING
rrNl rNJ
~Nili ('yN,J 'J
suitable for use in the manufacrure of parenteral
preparations.
IMPURITffiS
Otherdetectable impurities (thefallowing substances wauld, if
in the monograph. They are limited try thegeneral acaptance
0'?,J'" CI~CI °
HN
..-# CI CH3 a ¢II ~
NH
criterion for other/unspecified impurities and/or try the general o 0
monograph Substances/or pharmaceutical use (2034). It is
therefore not n«essary to identify these impurities for G. 7,7'-[ethane-I,I-diylbis [(2,3-dicWoro-4, I-phenylene)
demonstration of compliance. See also5.10. Control 0/ impurities piperazine-a, l-diylbutane-4, I-diyloxyIIdi(3,4-
in substances for pharmaceutical use) A, B, C, D, E, F, G. dihydroquinolin-2(1H)-one).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE«
A. 7-hydroxy-3,4-dihydroquinolin-2( IH)-one,
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2022 Articaine Hydrochloride 1-205
TESTS
Articaine Hydrochloride Solution S
(ph. EUT. monograph 1688) Dissolve 0.50 g in water R and dilute to 10 mL with the
same solvent.
than reference solution BY. (2.2.2, Method 1).
pH (2.2.3)
4.2 to 5.2.
320.8 23964-57-lJ 20.0 mL with the same solvent.
Action and use Related substances
Local anaesthetic. Liquid chromatography (2.2.29).
PhEu ~-_-----------
DEFINITION the mobile phase.
MethyI4-methyl-3-[[(2RS)-2-(propylamino) Reference solution (a) Dilute 1.0 mL of the test solution 10
propanoyl)amino]thiophene-2-carboxylate hydrochloride. 100.0 mL with the mobile phase. Dilute 1.0 mL of this
Content solution to 10.0 mL with the mobile phase.
98.5 per cent to 101.0 per cent (dried substance). Reference solution (b) Dissolve 5.0 mg of articaine
impurity A CRS and 2.5 mg of atticoine impurityE CRS in
CHARACTERS
Appearance phase. Dilute 1.0 mL of the solution to 50.0 mL with the
White or almost white, crystalline powder. mobile phase.
Solubility Column:
Freely soluble in water and in ethanol (96 per cent). nun,
- size: I ;;;; 0.25 m, 0 ;;;; 4.6
IDENTIFICATION - stationary phase: spherical end-capped oaadecylsi/yl silica gel
First identification: B, D. for chromatography R (5 urn);
Second idennfication: A, ,C, D. - temperature: 45 "C.
A. Dissolve 50.0 mg in a I gIL solution of hydrochlori< acidR Mobile phase Mix 25 volumes of aatonitri/e R and
and dilute to 100.0 mL with the same acid. Dilute 5.0 mL of 75 volumes of a solutionprepared as follows: dissolve 2.02 g
the solution to 100.0 mL with a I gIL solution of hydrochlori< of ,odium heptanesulfona" Rand 4.08 g of potassium
acid R. Examined berween 200 om and 350 om (2.2.25), the dihydrogen phospha" R in waW Rand dilute to 1000 mL with
solution shows an absorption maximum at 272 run. the same solvent. Adjust to pH 2.0 with phosphori< acid R.
The specific absorbance at the maximum is 290 to 320. Flow Ta,. I mUmin.
B. Infrared absorption spectrophotometry (2.2.24). Dueaion Spectrophotometer at 276 run.
Preparation Place dropwise 20 JlL of the test solution on Injection I0 ~L.
300 mg discs. Run time 5 times the retention time of articaine.
Test solution Dissolve 0.1 g in 5 mL of water R, add 3 mL Relauve retention Withreference to articaine (retention
of a saturated solution of sodium hydrogen carbonate Rand = =
shake twice with 2 mL of methylene chloride R. Combine the impurity E = about 0.86.
methylene chloride layers, dilute to 5.0 mL with methylene System suitability Reference solution (b):
chloride R and dry over anhydrous sodium sulfa" R. - resolution: minimum 1.2 between the peaks due to
Comparison articaine hydrochlori<le CRS. impurities A and E.
C. Thin-layer chromatography (2.2.27). Limits:
Test solution Dissolve 20 mg of the substance to be - impun'ty A: not more than the area of the corresponding
examined in 5 mL of ethanol (96 per cent) R. peak in the chromatogram obtained with reference
Reference soluuon Dissolve 20 mg of oniccine
hydrochloride CRS in 5 mL of ethanol (96 per cen!> R. - unspecified impun·ties: for each impurity, not more than the
Plate TLC sili", gelFm pIa,. R. with reference solution (a) (0.10 per cent);
Mobile phase uUthylamine R, ethylaceta,. R, heptane R - sum 0/impurities other thanA: not more than 5 times the
(10:35:65 VIVIV). area of the principal peak in the chromatogram obtained
Application 5 ~L. with reference solution (a) {0.5 per cent);
Development Overa path of 15 ern. - disregard limit. 0.5 times the area of the principal peak in
Drying In air.
(0.05 per cent).
principal spot in me chromatogram obtained with the
reference solution.
D. It gives reaction (a) of chlorides (2.3.1).
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1-206 Articaine Hydrochloride 2022

ASSAY
hydro,hlori< acid and 50 mL of ethanol (96 per ,en!! R. Carry
out a potentiometric titration (2.2.20) using 0.1 M sodium B. methyI4-methyl-3-[[(2RS)-2-[(I-methylethyl)arnino]
hydroxide. Read the volume added between the 2 points of propanoyl]amino]thiophene-2-catboxylale
inflexion. (isopropylaClieaine),
C"Ha.ClNaO,S.
STORAGE
IMPURlTlliS
Spedfi<d impurities A.
Otherdetectable impurities (thefollowing substances would, if F. 4-methyl-N-propyl-3-[[(2RS)-2-(propylarnino)propanoyl)
present at a sufficient level, be detected by oneor other of the tests amino]thiophene-2-carboxamide (articaine acid
in the monograph. They are limited by thegeneral a«ej>lanu propionamide),
criterion for olherlunspedfied impurities andlor by thegeneral
therefore not necessary It) idemify these impun"ries for
demonstration of compliance. See also5.10. Control of impurities
in substances for phormaceuncal use) B, C, D, E, F, G, H,
I,J.
G. methyl 3-[[(2RS)-2-(butylamino)propanoyI)amino]-4-
methylthiophene-2-carboxylate (butylarticaine),
A. methyI4-methyl-3-[[2-(propylamino)aeetyl]amino]
thiophene-2-carboxylate (acetamldoarticaine),
H~C H H-, CH3 H. methyl 3-[[(2RS)-2-(dipropylarnino)propanoyl]amino]-4-

s~N0~~c~ andenantiomer methylthiophene-2-earboxylate (dipropylartieaine),
'-"0 CH,
B. 4-methyl-3-[[(2RS)-2-(propylamino)propanoyl]amino]
thiophene-2-carboxylic acid (articaine acid),
1. methyl 3-amino-4-methylthiophene-2-eatboxylate
(3-aminoarticaine),
C. I-methylethyl 4-methyl-3-[[(2RS)-2-(propylamino)
H,C
,0&0 RY...
. s """'"
~_
Bf H
·CH J andenaotcmer
propanoyl]amino]thiophene-2-earboxylate (articaine ~ 0
isopropyl ester), CH,
J. methyl 3-[[(2RS)-2-bromopropanoyI)arnino)-4-
methylthiophene-2-earboxylate (bromo compound).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE.s
D. methyI3-[[(2RS)-2-(ethylamino)propanoyl]amino]-4-
methylthiophene-2-carboxylate (erhylarticaine),
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2022 Ascorbic Acid 1-207
TESTS
Ascorbic Acid Solution S
(ph. Eur. monograph 0253) Dissolve 1.0 g in carbon dioxide-free water Rand dilute £0
H Appearance of solution
HO~o Solution S is dear (2.2.1) and not more intensely coloured

than reference solution BY7 (2.2.2, Method ll).
HO OH
+ 20.5 to + 21.5.
176.1 5()'8I-7 same solvent.
Action and use Impurity E
Vitamin C. Maximum 0.2 per cent.
Preparations Test so/mum Dissolve 0.25 ginS mL of water R. Neutralise
Ascorbic Acid Injection using dilure sodium hydroxide solution R, then add 1 mL of
dilute acetic acid Rand 0.5 mL of calcium chlorick sohmon R.
Ascorbic Acid Tablets
Reference solution Dissolve 70 mg of oxalic acid R (dihydrate
Ascorbic Acid Chewable Tablets
of impurity E) in water R and dilute to 500 m.L with the
Paediatric Vitamins A, C and D Oral Drops same solvent; to 5 mL of the solution add 1 mL of dilute
Potassium Ascorbate Eye Drops acetic acid Rand 0.5 mL of calcium chloride solution R.
Vitamins Band C Injection Allow the solutions to stand for 1 h. Any opalescence in the
When Vitamin C is prescribed or demanded, Ascorbic Acid test solution is not more intense than that in the reference
shall be dispensed or supplied. solution.
Ph,,, _ Related substances
Liquid chromatography (2.2.29). Prepare the solucions
DEFINITION immediately before use.
(5R)-5- [(1 S) -1,2-Dihydroxyethylj-3,4-dihydroxyfuran-2 (5H)- Phosphate buffer solution Dissolve 6.8 g of potossivm
one. dilrydrogen phosphare R in waterfor chromatography Rand
Content dilute to about 175 mL with the same solvent. Filter through
99.0 per cent to 100.5 per cent. a membrane filter (nominal pore size 0.45 JJm) and dilute to
1000 mL with waterfor chromatography R.
CHARACTERS
Appearance
White or almost white, crystalline powderor colourless examined in the mobile phase and dilute to 10.0 mL with
crystals, becomingdiscoloured on exposure to air and me mobile phase.
moisture, Reference solution (a) Dissolve 10.0 mg of ascorbic acid
impurity C CRS in the mobile phase and dilute to 5.0 mL
Solubility
with the mobile phase.
Freelysoluble in water, sparingly soluble in ethanol
(96 per cent). Reference solution (b) Dissolve 5.0 mg of ascorbic acid
impurityD CRS and 5.0 mg of ascorbic acidCRS in the
mp
mobile phase, add 2.5 mL of reference solution (3) and
About 190 °C, with decomposition.
IDENTIFICATION Reference solution (c) Dilute 1 mL of the test solution to
First identification: B, C. 200 mL with the mobile phase. Mix I mL of this solution
Second identification: A, C, D. and 1 mL of reference solution (a).
A. Ultraviolet and visible absorption spectrophotometry Column:
(2.2.25). - size: 1=.0.25 m, 0 = 4.6 mm;
TestsoluMn Dissolve 0.10 g in water R and dilute - stationary phase: aminopropylsilyl silica gelfor
immediately to 100.0 mL with the same solvent. Add 1.0 mL chromatography R (5 urn);
of the solution to 10 mL of a 10.3 gIL solution of hydrochloric - temperature: 45°C.
acidR and dilute to 100.0 mL with water R. Mobi7e phase Phosphate buffer solution, acetonitrile Rl
Absorption maximum At 243 nm, determined immediately (25:75 VIV).
after dissolution. Flow rare 1.0 mUmin.
Spedfic absorbance ar the absorption maximum 545 to 585. Detection Spectrophotometer at 210 run.
B. Infrared absorption spectrophotometry (2.2.2if). Injection 20 ~ of the test solutionand reference
Comparison ascorbic acid CRS. solutions (b) and (c).
C. pH (2.2.3): 2.1 to 2.6 for solution S (see Tests). Run time 2.5 times the retention time of ascorbic acid.
D. To 1 mL of solution S add 0.2 mL of diltue nitric acidR Identification of impurities Use the chromatogram obtained
and 0.2 mL of silvernitrate solution R2. A grey precipitate is with reference solution (b) to identify the peaks due to
formed. impurities C and D.
Relativeretention With reference to ascorbic acid (retention
time = about 11 min): impurity D = about0.4;
=
impurity C about 1.7.
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1-208 Ascorbic Acid 2022
System suitability: criterion for otherlunspedfied impurities and/or by the general

- resolution: minimum 3.0 between the peaks due to monograph Substances for pharmaceuucal use (2034). It is
ascorbic acid and impurity C in me chromatogram therefore not necessary to identify these impuriues for
obtained with reference solution (c); demonstration of compliance. See also 5.10. Control of impurities
- signal-co-noise ratio: minimum 20 for the peak due to in substances for phannaceutical use) A, F, G} H.
impurity C in the chromatogram obtained with reference
solution (b).
(yCHO
Limits:
- impuniies C, D: for each impurity, not more man
1.5 times the area of the corresponding peak in the A. furan-2-carbaldehyde,
(0.15 per cent);
- unspecified impwitks: for each impurity) not more than the H~H
area of the peak due to ascorbic acid in the chromatogram HO') H;"\OH 0
obtained with reference solution (b) (0.10 per cent); HO
- sum of impun"ties otherthan C and D: not more than twice
the area of the peak due to ascorbic acid in the
C. L-xy/o-hex-2-ulosonic acid (t-scrbosonic acid),
HOH
y;c
(0.2 per cent); o DCH,
ascorbic acid in the chromatogram obtained with H '
reference solution (b) (0.05 per cent). HO···· H/OH 0
Copper HO
Maximwn 5 ppm.
Atomic absorption spectrometry (2.2.23, Melhod/). D. methyl L-xylo-hex-2-ulosonate (methyl t-sorbosonate),
Test solulUm Dissolve 2.0 g in 0.1 M nitric acidand dilute to
25.0 mL with the same acid
Reference solutions Prepare the reference solutions (0.2 ppm,
0.4 ppm and 0.6 ppm) using cqpper standard solution (10 ppm E. oxalic acid,
Gu) R, diluting with 0.1 M nitric add.
Source Copper hollow-cathode lamp.
Wavelength 324.8 om.
Atomisation deoice Air-acetylene flame.
H°1:<O HO OH
Adjust the zero of the apparatus using 0.1 M ninic acid.
Iron
F. (5R) -5- [( 1R)-I ,2-dihydroxyethyl]-3,4-dihydroxyforan-2
Maximwn 2 ppm.
(5H)-one,
Atomic absorption spectrometry (2.2.23, Method /).
Test solution Dissolve 5.0 g in 0.1 M nitric acid and dilute to H
Reference solutions Prepare the reference solutions (0.2 ppm,
H~~O
0.4 ppm and 0.6 ppm) using iron standard solution (20 ppm
Fe) R, diluting with 0.1 i\1 nimc acid. HO OH
Source Iron hollow-cathode lamp.

G. (R)-[(2R)-3,4-dihydroxy-5-oxo-2,5-dihydrofuran-2-yl]
hydroxyacetic acid)
Adjust the zero of the apparatus using 0.1 M nitric acid. 0
H
1:<
Sulfated ash (2.4.14) .OH
H3CO .'0
Maximum 0.1 per cent, determined on 1.0 g. 0
ASSAY
Dissolve 0.150 g in a mixture of 10 rnL of dilute sulfuric HO OH
acid Rand 80 mL of carbon dioxide-free water R. Add I rnL of
starch solution R. Titrate with 0.05 M iodine until a persistent H. methyl (R)-[(2R)-3,4-dihydtoxy-5-oxo-2,5-dihydrofuran-
violet-blue colour is obtained. 2-yl]hydroxyacetate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pl>E<r
I rnL of O. 05 M iodine is equivalent to 8.81 mg of C.,HaO•.
STORAGE
In a non-metallic container, protected from light.
IMPURITIES
Specified impurities G, D, E.
Otherdetectable impurities (the fol/Qwing substances would, if
present at a sufficient level, be detected by oneor otherof the tests
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2022 Asparagine 1-209
Ascorbyl Palmitate *** Asparagine Monohydrate

** **
(Ph. Eur. monograph 0807) ***** (Ph. Eur. monograph 2086)
0
H'C~~ 14 0 0
0
H,NV::'" • H,o
HO OH C,H"N,O"H,O 150.1 5794-13-8
c"H38O, 414.5 137-66-6 Action and use

Amino acid.
Action and use
Pl>Ev _
Excipient.
PI>£v _ DEFINITION
(2S)-2,4-Diamino-4-oxobutanoic acid monohydrate.
DEFINITION
(2S)-2-[(2R)-3,4-Dihydroxy-5-oxo-2,5-dihydrofuran-2-yl]-2- Content
hydroxyethyl hexadeeanoate. 99.0 per cent to 101.0 per cent (dried substance).
Content CHARACTERS
98.0 per cent to 100.5 per cent (dried substance). Appearance
CHARACTERS
crystals.
Appearance
White or yellowish-white powder. Solubility
Slightly soluble in water, practically insoluble in ethanol
Solubility
(96 per cent) and in methylene chloride.
Practically insoluble in water, freely soluble in ethanol
(96 eec cent) and in methanol, practically insoluble in IDENTIFICATION
methylene chloride and in fatty oils. Fjrst identification: A J BJ D.
IDENTIFICATION Second identification: A, C, D.
A. Specific optical rotation (see Tests). A. Specific optical rotation (see Tests).
B. Infrared absorption spectrophotometry (2.2.24). B. Infrared absorption spectrophotometry (2.2.24).
Comparison ascorbylpalmitate CRS. Camparison asparagine monohydrate CRS.
C. Dissolve about 10 mg in 5 mL of methanol R. C. Thin-layer chromatography (2.2.27).
The solution decolourises dkhlorophenoh"ndvphenol standard Testsolution Dissolve 10 mg of the substance to be
solution R. examined in water R and dilute to 10 mL with the same
TESTS solvent.
Soludon S Reference solution Dissolve 10 mg of asparagine
Dissolve 2.50 g in methanol R using sonication and dilute to monohydrate CRS in water R and dilute to 10 mL with the
25.0 mL with the same solvent. same solvent.
Appearance of solution Plate TLC silirogel plate R.
Solution S is clear (2.2.1) and not more intensely coloured Mobile phase glacial acetic acid RJ water R, butanol R
than reference solution BY4 (2.2.2, iWethod l). (25:25:50 VIVIV).
Specific optical rotation (2.2.7) Application 5 IlL.
+ 21 to + 24 (dried substance), determined on solution S. Development Over 2/3 of the plate.
Related substances Drying At 110 °C for 15 min.
The thresholds indicated underRelated substances Detection Spray with ninhydrin solution R andheat at 105°C
(Table 2034.-1) in the general monograph Substances for for 10 min.
pharmaceutical use (2034) do not apply. Results The principal spot in the chromatogram obtained
Loss on drying (2.2.32) with the test solution is similar in position, colour and size to
Maximum 1.0 per cent, determined on 1.000 g by drying in the principal spot in the chromatogram obtained with the
VlUUO at 60°C for 5 h. reference solution.
Sulfated ash (2.4.14) D. Loss on drying (see Tests).
Maximum 0.1 per cent, determined on 1.0 g. TESTS
ASSAY Solution S
Dissolve 0.200 g in 50 mL of ethanol (96 per cent) R. Dissolve with heating 2.0 g in carbon dioxide-free water Rand
Add 30 mL of water R and titrate with 0.05 M iodine until a dilute to 100 mL with the same solvent.
yellow colour is obtained. Appearance of solution
I mL of 0.05 M iodine is equivalent to 20.73 mg of Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
C22H3S07· pH (2.2.3)
STORAGE 4.0 to 6.0 forsolution S.
In an airtight container, protected from light. Specific optical rotation (2.2.7)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>£II + 33.7 to + 36.0 (dried substance).
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1-210 Asparagine 2022
Dissolve 2.50 g in a 309.0 gIL solution of hydrochloric acid R Chlorides (2.4.4)

and dilute to 25.0 mL with me same acid. Maximum 200 ppm.
Related substances Dilute 12.5 mL of solution S to 15 mL with waler R.
liquid chromatography (2.2.29). Prepare the solutions Sulfates (2.4.13)
immediately before use. Maximum 200 ppm.
Test solution Dissolve 0.100 g of the substance to be To 0.75 g add 2.5 mL of dilute hydrochlori< acidR and dilute
examined in water R and dilute to 10.0 mL with the same to 15 mL with distilled water R. Examine after 30 min.
solvent.
Ammonium (2.4.1, Method B)
Reference solution (a) Dilute 1.0 mL of the test solution to Maximum 0.1 per cent, determined on 10 mg.
Prepare me standard using 0.1 mL of ammonium standard
Reference solution (b) Dilute 1.0 mL of reference solution (a) solution (100 ppm NH.) R.
to 10.0 mL with water R.
Iron (2.4.9)
Reference solution (c) Dissolve 5.0 mg of aspartic acid R Maximum 10 ppm.
(impurity A) in water R and dilute to 10.0 mL with the same
solvent. Dilute 1.0 mL of the solution to 10.0 mL with Dissolve 1.0 g in dilute hydrochloric acidR and dilute to
water R. 10 mL with the same acid. Shake 3 times with 10 rnL of
methylisobutyl ketone Rl for 3 min. Wash the combined
Reference solution (d) Dissolve 3.0 mg of asparagine organic phases with 10 mL of water R for 3 min.
impurity C CRS in 40 mL of the mobile phase using The aqueous phase complies with the limit test for iron.
sonication and dilute to 50.0 mL with the mobile phase.
Dilute 1.0 mL of the solution to 10.0 mL with water R. Loss on drying (2.2.32)
Reference solution (e) Mix 5 mL of reference solution (c)
with 25 mL of reference solution (a) and dilute to 10 mL
with water R. Sulfated ash (2.4.14)
Column:
- size: 1= 0.25 m, 0 ;;;; 4.6 mm; ASSAY
- stationary phase: end-capped ocradecylsiiyl silica gelfor Dissolve 0.110 gin 5 mL of anhydrous formic acid R.
chromatography R (5 urn); Add''lO' mr;-of anhydrous acetic acid R. Titrate with 0.1 M
- temperature: 25°C. perchbJric acid, determining the end-point potentiometrically
Mobile phase Dissolve 13.6 g of potassium dihydrogen (2.2.2fJ).
phosphate R and 2.16 g of sodium oaanesulfonau R in about I mL of 0.1 M pcrchloric acidis equivalent to 13.21 mg
900 mL of waterfor chromaragraphy R. Adjust to pH 2.2 with of C.HaN,O,.
phosphori< acidR and dilute to 1000 mL with waterfor
IMPURITIES
chromatography R. Add 5 mL of ace",nitrile Rl.
Specified impun·ties A, C.
Otherdetecrable impurilies (thefollowing subsrances would, if
Detection Spectrophotometer at 210 nm. present at a sufficient leveJ~ be detected by oneor other oj the tests
Injection 20 pL. in the monograph. They are limited by thegeneral acceptance
Run time Twice the retention time of asparagine. criterion for other/unspecified impurities and/or by the general
Identification of impurities Use the chromatogram obtained monograph Sub,rances for pharmaceutical use (2034). II is
with reference solution (c) to identify the peak due to therefore not necessary to identify these impurities Jor
impurity A; use the chromatogram obtained with reference demonstration of compliance. See also 5.10. Control of impurities
solution (d) to identify the peak due to impurity C. in mbstanus Jor pharmaceutical use) B~ D~ E~ F~ G~ H.
Relativeretention With reference to asparagine (retention
time ;;;: about 6.6 min): impurity C = about 0.6; .
System suitability Reference solution (e):
- resolution: minimum 5.0 between the peaks due to A. (2S)-2-aminobutanedioicacid (aspartic acid),
asparagine and impurity A.
H NH,
H02C~CO:zH
- for impurity A, use the concentration of impurity A in
- for impurity C, use the concentration of impurity C in B. (2S)-2-aminopentanedioic acid (glutamic acid),
reference solution (d);
- for impurities other than A and C, use the concentration o
Jl.~ .NH,
of asparagine monohydrate in reference solution (b). o HN" T I(
Limits:
H,N-
....l J..
~-U
.NH a
- impun·ty A: maximum 0.5 per cent,
~ ;mpun·cy C: maximum 0.1 per cent, o
- unspecified impurities: for each impurity, maximwn
0.05 per cent, C.2,2'-(2EJ5E)-3,6-dioxopiperazine-2,5-diyl]diacetamide,
- roUJ1: maximum 0.8 per cent;
D. (2E)-but-2-enedioic acid (fumaric acid),
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2022 Aspartame 1-211
Solubility
Sparingly soluble or slightly soluble in water and in ethanol
(96 per cent), practically insoluble in hexane and in
methylenechloride.
E. (2S)-2,5-diamino-5-oxopentanoic acid (glutamine), IDENTIFICATION
(2.2.25).
Test solurion Dissolve 0.1 g in ethanol (96 per cem) R and
F. (2S)-2-[[(2S)-2,4-diamino-4-oxobutanoyl]amino] dilute to 100 mL with the same solvent.
butanedioic acid (asparaginylaspartic acid), Spectral range 230-300 om.
Absorption maxima At 247 nm, 252 nm, 258 nm and
264 om.
Preparation Discs.
Comparison aspartame CRS.
G. (2S)-4-amino-2-[[(2S)-2-amino-3-carboxypropanoyl] examined in 2.5 mL of water R and dilute to 10 mL with
amino]-4-oxobutanoic acid (a-aspartylasparagine), acetic acid R.
Reference solution Dissolve 15 mg of aspartame CRS in
2.5 ml; of water R and dilute to 10 ml, with acetic acid R.
Pkue TLC silica gel G plare R.
Mobl1e phase water R, anhydrous formic acid R, methanol R,
methylene chloride R (2:4:30:64 VIVIVIJI).
Application 20 ~L.
H. (2S)-4-amino-2-[[(2S)-2,4-diamino-4-oxobutanoyl] Development. Overa path of 15 em.
amino]-4-oxobutanoic acid (asparaginylasparagine). Drying In air.
_ _ _~ PbE" Detection Spraywithninhydn·n solution R and heat at
100-105 'C for IS min.
Results The spot in the chromatogram obtainedwith the
test solution is similar in position, colour and size to the spot
Aspartame in the chromatogram obtainedwith the reference solution.
D. Dissolve about 20 mg in 5 ml, of melhanol R and add
(Ph. Bur. monograph 0973) 1 mL of alkaline hydroxylamine solution Rt. Heat on a water-
bath for 15 min. Allow to cool and adjust to about pH 2
~
with dilute hydrochloric acidR. Add 0.1 mL of/eme chloride
solution Rl, A brownish-red colouris produced.
o H TESTS
H~ .Jl N '. O....CH
Y. 'H 3
Solution S
H / 0
Ho,c 100 ml, with the same solvent.
294.3 22839-47-0 Solution S'is clear (2.2.1) and not more intensely coloured
than reference solution GY, (2.2.2, Merhod Il).
Action and use Conductivity (2.2.38)
Sweetening agent. Maximum 30 IlS·cm-1.
PbE" _ Dissolve 0.80 g in carbon dioxide-free waterR prepared from
distil/ed waterR and dilute to 100.0 ml, with the same
DEFINITION solvent. Measure the conductivity of the solution (C1) and
(3S)-3-Amino-4-[[(2S)-I-methoxy-l-oxo-3-phenylpropan-2- that of the water used for preparing the solution (C2 ) .
yl]amino]-4-oxobutanoic acid (methyl c-t-aspanyl-r- The readings must be stable within 1 per cent over a period
phenylalaninate). of30 s.
Content Calculate the conductivity of the solution of the substance to
98.0 per cent to 102.0 per cent (dried substance). be examinedusing the foHowing expression:
CHARACTERS
Appearance
C, - 0.992 C,
White or almost white, slightly hygroscopic, crystalline
powder.
+ 14.5 to + 16.5 (dried substance).
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1-212 Aspartic Acid 2022
Dissolve 2.00 g in a 690 gIL solution of anhydrous formic 1 mL of 0.1 At1 perchlotic acid is equivalent to 29.43 mg
acidR and dilute to 50.0 mL whh the same solution. of C14HlSNzOs.
Measure within 30 min of preparation. STORAGE
Related substances In an airtight container.
IMPURITIES
examined in a mixture of 1.5 volumes of glacial acetic add R
Specified impurities A, G.
and 98.5 volumes of water R and dilute to 100.0 mL with Otherdet«table impurities (thefollowing sub'tances would, if
the same mixture of solvents, presen; at a sufficient level be detected by oneor other of the tests
J
Referenee ,0/ution (a) Dissolve 4.5 mg of aspartame

in the monograph. They are limited by thegeneral a«eptance
criterion far otherlumpe<ified impurities and/ar lry the general
impurity A CRS in a mixture of 1.5 volumes of glacial acetic
monograph Substances far pharmaceutical use (2034). It is
acid Rand 98.5 volumes of water R and dilute to 50.0 mL
therefore not necessary to identify these impuniies for
with the same mixture of solvents.
Reference ,oIuu'on (b) Dissolve 30.0 mg of phenylalanine R in substances for phannauutiCflI use) B.
(impurity C) in a mixture of 15 volumes of glacial acetic
acid Rand 85 volumes of waterR and dilute [0100.0 mL
° NH
ut
with the same mixture of solvents. Dilute 1.0 mL of this
, ;
solution to 10.0 mL with water R.
Reference so/utien (c) Dilute 5.0 mL of the test solution to ~ HN~C~H
10.0 mL with water R. Dilute 3.0 mL of this solution to o
Ref.....a ,0/ution (d) Dissolve 30.0 mg of L-aspa~-L A. 2-[(2S,5S)-5-benzyl-3,6-rlioxopiperazin-2-yl]acetic acid,
phenylalanine R (impurity B) in a mixture of 15 volumes of
glacial acetic acidR and 85 volumes of water R and dilute to
100.0 mL with the same mixture of solvents. Dilute 1.0 mL
of the solution to 10.0 mL with water R. Mix 1.0 mL of this
solution with 1.0 mL of reference solution (b).
Column
- size: I ::;; 0.25 m, 0 ::;; 4.0 nun;
- stationary phase: oaode<y/sjlyl silica gelfar chramatagraphy R
(5-10 pm).
MobUe phase Mix 10 volumes of aa",nitrile Rand B. (3S)-3-amino-4-[[(IS)-I-carboxy-2-phenylethyl]amino]-4-
90 volumes of a 6.8 gIL solution of potassium dihydragen oxobutanoic acid (o-t-aspartyl-t-phenylalanine),
phosphate R previously adjusted to pH 3.7 with phosphoric
acidR.
Flow rate I mUmin.
Injection 20 IlL. C. (2S)-2-amino-3-phenylpropanoic acid (t-phenylalanlne).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Run time Twice the retention time of aspartame.
SYStem suitability Reference solution (d):
impurities Band C.
Limits: Aspartic Acid
- impurity A: not more than the area of the principal peak
in the chromatogram obtained with reference solution (a) (Ph. Bur. monograph 0797)
(1.5 per cent);
H NHz
- impurity C: not more than me area of the principal peak
Ho,c~ CD,H
(0.5 per cent);
- sumof impun·ties other thanA and C: not more than the 133.1 56-84-8
with reference solution (c) (1.5 per cent); Action and use
- disregard limit: disregard any peak due to the solvent. Amino acid.
Loss on drying (2.2.32) PhE" _
an oven at 105°C. DEFINITION
Sulfated ash (2.4.14) (2S)-2-Aminobutanerlioic acid (r-aspartic acid).
Maximum 0.2 per cent, determined on 1.0 g. Content
ASSAY 98.5 per cent to 101.5 per cent (dried substance).
Dissolve 0.250 g in 1.5 mL of anhydrous formic acid Rand CHARACTERS
60 mL of anhydrous acetic acid R. Titrate inunerliately with Appearance
0.1 M perchloric acid, determining the end-point White or almost white, crystalline powderor colourless
potentiometrically (2.2.20). crystals.
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2022 Aspartic Acid 1-213
Solubility Mobl1e phase 2-propanol R, 0.5 gIL solution of copper sulfate

Slightly soluble in water, practically insoluble in ethanol pemahydrate R (5:95 VIII).
(96 per cent). It dissolves in dilute mineral acids and in Flow rate 1.0 mIJmin.
dilute solutions of alkali hydroxides.
IDENTIFICATION Injection 20~.
Piru identification {cany out either tests A, C or tests C, D.}
Relative retention Wilh reference to aspartic acid (retention
Secondidentification: A, B, E. time = about 12 min); impurity I = about 0.85.
A. Specific optical rotation (2.2.7): + 24.0 to + 26.0 (dried Systemsuitability Reference solution (b):
substance). - resolution: minimum 2.0 between the peaks due to
Dissolve 2.00 g in hydro<hloric acidRt and dilute to 25.0 mL impurity I and aspartic acid.
with the same acid. Calculation of percentage amtent:
B. A suspension of 1 gin 10 mL of water R is strongly acid - for impurity I, use the concentration of impurity I in
(2.2.4). reference solution (c).
C. Infrared absorption spectrophotometry (2.2.24). Limit:
Comparison aspartic acid CRS. - impun·ty 1: maximum 0.3 per cent.
D. Enantiomeric purity (see Tests). Other dlcarboxylic acids
E. Thin-layer chromatography (2.2.27). Liquid chromatography (2.2.29).
Test soluuon Dissolve 10 mg of the substance to be Test solutitm Dissolve 0.500 g of the substance to be
examined in 2 mL of dilute ammonia Rl and dilute to 50 mL examined in 2.0 mL ofa 618 gIL solution of hydrochloric
with water R. acidR and dilute to 10.0 mL with water R.
Reference solution Dissolve 10 mg of aspartic acidCRS in Re/erenasolution (a) Dissolve 20.0 mg of malicacidR
2 mL of dilute ammoniaRl and dilute to 50 mL with (impurity A) in water R and dilute to 20.0 mL with the seme
water R. solvent.
Plate TLC silica gelpkue R. Reference solution (b) Dissolve 10.0 mg of maleic acidR
(impurity H) in water R and dilute to 10.0 mL with the same
lHobi/e phase glacial acetic acidR, water R, butanol R
solvent. Dilute 1.0 mL of the solution to 10.0 mL with
(20:20:60 VIVIV).
water R.
Application 5 ~L.
Reference solution (c) Dilute 1.0 mL of reference solution (b)
Development Over 2/3 of the plate. to 10.0 mL with reference solution (a).
Drying In air. Reference solution (d) Dilute 1.0 mL of reference solution (a)
Detection Spray with ninhydn"n solution R and heat at 105 DC to 10.0 mL with waterR.
for 15 min. Reference solution (e) Dissolve 10.0 mg of/umaric acidR
Results The principal spot in the chromatogram obtained (impurity B) in water R and dilute to 10.0 mL with the same
with the test solution is similar in position, colour and size to solvent. Dilute 1.0 mL of me solution to 100.0 mL with
the principal spot in the chromatogram obtained with the water R.
reference solution. Column:
TESTS - size: 1= 0.30 m, 0 = 7.8 mm;
Appearance of soludon - stationary phase: cation-exchange resin R (9 um),
The solution is clear (2.2.1) and not more intensely coloured - temperature: 30 DC.
than reference solution BY6 (2.2.2, lHemod If). Mobile phase 0.39 gIL solution of sulfuric acidR.
Dissolve 0.5 g in a 103 gIL solution of hydro<h/oric acid Rand Flow rat< 0.6 mllmin.
dilute to 10 mL with the same acid. Detection Spectrophotometer at 214 nm.
Enantlomerlc purity Injection 10 ~L.
Liquid chromatography (2.2.29). Run time 4 times the retention time of impurity H.
Test solution Dissolve 0.100 g of the substance [Q be Identification of impurities Use the chromatogram obtained
examined in waterR and dilute to 100.0 mL with the same with reference solution (c) to identify the peaks due to
solvent. impurities A and H; use the chromatogram obtained with
Reference solution (a) Dissolve 0.1 00 g of n-aspamc acid R reference solution (e) to identify the peak due to impurity B.
(impurity I) in water R and dilute to 100.0 mL with the same Relative retention With reference to impurity H (retention
solvent. = =
time about 7.5 min); impurity A about 1.2;
Reference solution (b) Dissolve 0.100 g of the substance to impurity B = about 2.0.
be examined in 90 mL of water R, add 0.3 mL of reference System suitability Reference solution (c):
solution (a) and dilute to 100.0 mL with water R. - resolution: minimum 1.5 between the peaks due to
Reference solution (c) Dilute 0.3 mL of reference solution (a) impurities Hand A.
to 100.0 mL with water R. Calculation of percentage contents:
Column: - for impurity A, use the concentration of impurity A in
=
- size: 1 = 0.15 m, 0 4.6 mm; reference solution (d);
- stationary phase: L-penidOamine coated silica gelfor chirat - for impurities Band H, use the concentration of
separations R (5 ~m); impurity H in reference solution (b)j
- temperature: 30 DC. - for impurities other than Band H, use the concentration
of impurity A in reference solution (d).
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1-214 Aspartic Acid 2022
Limits: - tetal: maximum 1.0 per cent;

- impun"ty A: maximum 0.2 per cent; - reporting threshold: 0.05 per cent,
- ;mpun"IY B: maximum 0.10 per cent; Chlorldea (2.4.4)
- impun"IY H: maximum 0.10 per cent; Maximum 200 ppm.
0.10 per cent; Dissolve 0.25 g in 3 mL of dilute nitric acid R and dilute to
15 mL with wa/erR. Add I mL of wa/er R instead of I rnL
of dilute nitric add R.
Sulfates (2.4. H)
Maximum 300 ppm.
Amino acid analysis (2.2.56). For analysis, use Method 1.
Dissolve 0.5 g in 4 rnL of hydrochloric acidR and dilute to
The concentrations of the test and reference solutions may
be adapted according to the sensitivity of the equipment 15 mL with distilled water R. Carry out the evaluation of the
test after 30 min.
used. The concentrations of all solutions are adjusted so that
the systemsuitability requirements descnbed in general Anunonlum
chapter 2.2.46 arefulfilled, keeping the ratios of Amino acid analysis (2.2.56) as described in the test for
concentrations between all solutions as described. ninhydrin-positive substances with the following modification.
Solution A dIlute hydrochloric acid RJ or a samplepreparation Injection Test solution, reference solution (c) and blank
buffer suitable for the apparatus used. solution.
Test solution Dissolve 30.0 mg.of the substance to be Limit:
examined in solution A and dilute to 50.0 mL with - ammonium at 570 nm: not more than the area of the
solution A. corresponding peak in the chromatogram obtained with
Reference solution (a) Dilute 1.0 mL of the test solution to reference solution (c) (0.02 per cent), taking into account
100.0 mL with solution A. Dilute 2.0 rnL of this solution to the peak due to ammonium in the chromatogram
10.0 rnL with solution A. obtainedwith the blanksolution.
Reference solution (b) Dissolve 30.0 mg of proline R in Iron (2.4. 9)
solution A and dilute to 100.0 mL with solution A. Dilute Maximum 10 ppm.
1.0 mL of this solution to 250.0 mL with solution A. In a separating funnel, dissolve 1.0 g in 10 rnL of dilute
Reference solution (c) Dilute 6.0 rnL of ammonium standard hydrochloric acid R. Shake with 3 quantities, each of 10 nIL,
,olution (100 ppm NH.,) R to 50.0 rnL with solution A. Dilute of methyl isobutyl ketone Rl, shaking for 3 min each time.
1.0 rnL of this solution to 100.0 rnL with solution A. To the combined organic layers add J0 mL of water Rand
shakefor 3 min. Use the aqueous layer.
Reference solution (d) Dissolve 30 mg of isoleucine Rand
30 mg of leucine R in solution A and dilute to 50.0 rnL with Loss on drying (2.2.32)
solution A. Dilute 1.0 rnL of the solution to 200.0 rnL with Maximum 0.5 per cent, determined on 1.000 g by drying in
solution A. an oven at 105°C.
Reference solution (e) Dissolve 30.0 mg of alanine R Sulfated ash (2.4.14)
(impurity D), 60.0 mg of asparagine R (impurity G) and Maximum 0.1 per cent, determined on 1.0 g.
30.0 mg of glulllmic acidR (impurity C) in solution A and ASSAY
dilute to 100.0 rnL with solution A. Dilute 1.0 rnL of the
Dissolve 0.100 g in 50 rnL of carbon dioxide-free wa/erR, with
solution to 250.0 mL with solution A. slight heating if necessary. Cool and titrate with 0.1 M sodium
Blank solution Solution A. hydroxide determining the end-pointpotentiometrically
Inject suitable, equal amounts of the test solution, blank (2.2.211).
solutionand reference solutions (a), (b), (d) and (e) into the I mL of 0.1 M sodium hydroxide is equivalent to 13.31 mg of
amino acid analyser. Run a program suitable for the C.H,NO•.
determination of physiological amino acids.
STORAGE
System suitabi/if)! Reference solution (d):
- resolution: minimum 15 between the peaks due to
isoleucine and leucine. IMPURITIES
Calculation of percentage contents: Specified ;mpun'iies A, B, C, D, H, G, 1.
- for impurities C, D and G, use the concentration of each Otherdetectable impuritks (thefollowing subslances WQuld, if
impurity in reference solution (e)j present at a sufficient level, be deteeted hy oneor other of the tests
- for any ninhydrin-positive substance detected at 570 run, in the monograph. They are limited hy thegeneral acceptance
use the concentration of aspartic acid in reference criterion for other/unspecified impurities andJor by the general
solution (a); monograph Sub'lIlnces for pharmaceutical use (2034). It is
- for any ninhydrin-positive substance detected at 440 run, therefore no! necessary to identify these impuritks for
use the concentration of proline in reference solution (b); demonstration of compliance. See also5.10. Control of impuniies
if a peak is above the reporting threshold at both in substanas for pharmaceutical use) E) F.
wavelengths, use the resultobtained at 570 nm for
HO H
quantification.
H~CY andeoanncmer
Limits: co,H
- impurities C, D, G: for each impurity, maximum
0.2 per cent; A. (2RS}-2-hydroxybutanedioic acid (malic acid),
- any ninhydnn-posiuVe substance: for each impurity,
maximwn 0.10 per cent;
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2022 Aspirin 1-215
Ph", _
DEFINITION
B. (2E)-but-2-enedioic acid (fumaric acid), 2-(Acetyloxy)benzoic acid.
Content
CHARACTERS
C. (2S)-2-aminopentanedioic acid (glutamic acid), Appearance
crystals.
Solubility
Slightly soluble in water, freely soluble in ethanol
D. (2S)-2-aminopropanoic acid (alanine), (96 per cent).
Ho,C~Co,H mp
About 143°C (instantaneous method).
E. butanedioic acid (succinic acid), IDENTIFICATION
Second idendficotion: B, C, D.
Comparison autylsalicyli< acid CRS.
F. (2S)-2,5-diamino-5-oxopentanoic acid (t-glutamine), B. To 0.2 g add 4 mL of dilute sodium hydroxide solution R
and boil for 3 min. Cool and add 5 mL of dilute sulfuric
o H N~ acid R. A crystalline precipitate is formed, Filter, wash the
H;zN~C02H precipitate and dry at 100-105 "C. The melting point
(2.2.14) is 156 "C to 161 "C.
G. (2S)-2,4-diamino-4-oxobutanoic acid (asparagine), C. In a [est tube mix 0.1 g with 0.5 g of calcium hydroxide R.
Heat the mixture and expose to the fumes produced a piece
("'CO,H of filter paper impregnated with 0.05 mL of nitrobenzaldehyde
solution R. A greenish-blue or greenish-yellow colour develops
Co,H
on the paper. Moisten the paper with dilute. hydrodloric
H. (2Z)-but-2-enedioic acid (maleic acid), acid R. The colourbecomes blue.
D. Dissolve with heating about 20 mg of the precipitate
obtained in identification test Bini0 mL of water Rand
cool. The solution gives reaction (a) of salicylates (2.3.1).
TESTS
I. (2R)-2-aminobutanedioic acid (n-aspartic acid). Appearance of solution
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf"
Method II).
Dissolve 1.0 g in 9 mL of ethanol (96 per cent) R.
Related substances
Aspirin Liquid chromatography (2.2.29). Prepare the solutions
(Awy/salicylic Acid, Ph. Eur, monograph 0309)
examined in acetonirn7e for chromatography R and dilute to
Reference solution (a) Dissolve 50.0 mg of salicylic add R
(impurity C) in the mobile phase and dilute to 50.0 mL with
the mobile phase. Dilute 1.0 mL of the solution to 100.0 mL
180.2 5{}-78-2
Reference solution (b) Dissolve 10 mg of salicyli< acidR
Action and use (impurity C) in the mobile phase and dilute to 10.0 mL with
Salicylate; non-selective cycle-oxygenase inhibitor; the mobile phase. To 1.0 mL of the solution add 0.2 mL of
antipyretic; analgesic; anti-inflammatory. the test solution and dilute to 100.0 mL with the mobile
phase.
Preparations
Aspirin Tablets Reference solution (c) Dissolve with the aid of ultrasound the
contents of a vialof acetylsalkylk acid forpeak
Aspirin Dispersible Tablets identification CRS (containing impurities A, B, D, E and F)
Aspirin Effervescent Soluble Tablets in 1.0 mL of acetomtrile R.
Aspirin Gastro-resistant Tablets Column:
Aspirin and Caffeine Tablets - size: 1= 0.25 m, 0 = 4.6 mm;
Co-codaprin Tablets - stationary phase: oClad«ylsi/y1 silica gelfor chromatography R
Co-codaprin Dispersible Tablets (5 pm).
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1-216 Aspirin Lysine 2022
Mobile phase phosphotic acidR, acetonitrile for

chromatography R, waterR (2:400:600 VIVW).
Flow rate 1 mUmin.
Detection Spectrophotometer at 237 nm. B. 4-hydroxybenzene-I,3·dicarboxylic acid
Injection 10 ~L. (4-hydroxyisophthalic acid),
Run time 7 times the retention time of acetylsalicylic acid.
Co,H
Identification of impuruies Use the chromatogram obtained
impurity C; use the chromatogram supplied with
CX "'- OH
aatyualicylic acidfor peak identification CRS and the

C. 2-hydroxybenzenecarboxylic acid (salicylic acid),
chromatogram obtainedwith reference solution (c) to identify
the peaks due to impurities A, B, D, E and F.
Relative retention With reference to acetylsalicylic acid
(retention time = about 5 min): impurity A = about 0.7;
impurity B ;;;; about 0.8; impurity C = about 1.3;
impurity D = about 2.3; impurity E = about 3.2;
impurity F = about 6.0.
System suitabtlity Reference solution (b):
- resolution: minimum 6.0 between the peaks due to D. 2-[[2-(acetyloxy)benzoyl)oxy)benzoic acid
acetylsalicylic acid and impurity C. (acetyl salicylsalicylic acid),
Limits:
(0.15 per cent);
- unspecified impurities: for each impurity) not more than
E. 2-[(2-hydroxybenzoyl)oxy]benzoic acid (salsalate,
(0.05 per cent);
- total: not more than 2.5 times the area of the principal salicylsalicylic acid),
- disregard limit. 0.3 times the area of the principal peak in
(0.03 per cent).
Maximum 0.5 per cent) determined on 1.000 g by drying in
vacuo.
Sulfated ash (2.4.14) F. 2-(acetyloxy)benzoic anbydride (acetylsalicylic anbydride).
Maximum 0.1 per cent, determined on 1.0 g. ___________________ ''bE"
ASSAY
In a flask with a ground-glass stopper, dissolve 1.000 g in
10 rnL of ethanol (96 pereen\! R. Add 50.0 rnL of 0.5 M
sodium hydroxide. Close the flask and allow to stand for 1 h. Aspirin Lysine ***
Using 0.2 mL of phenolphthalein solution R as indicator, titrate *** ***
with 0.5 M hydrochloric acid. Carry out a blank titration. (DL-Lysine Aatyualicy/ate, Ph. Bur. monograph 2812) ***
C.H.O,. H NH2
STORAGE H,N~co,H
In an airtight container. andeoanliomer
IMPURITIES
Specified impurities A, B, C, D, E, F.
326.3 62952-06-1
I""'yCo,H
Action and use
HO~ Salicylate; non-selective cyclo-oxygenase inhibitor;
antipyretic; analgesic; anti-inftanunatory.
A. 4-hydroxybenzoic acid, PIlE" _
DEFINITION
(2RS)-2,6-Diaminohexanoic acid 2-(acetyloxy)benzoate.
Content
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2022 Aspirin Lysine 1-217
CHARACTERS Mobile phase:

Appearance - mobile phaseA: 0.55 gIL solution of sodium
White or almost white, hygroscopic, crystalline powder. oaonesulfonate R adjusted to pH 2.7 with phosphoric
Solubility acid R;
Very soluble in water, slightly soluble in ethanol - mobile phaseB: acetonitrile Rt,
(96 per cent), practically insoluble in heptane.
IDENTIFICATION (min) (per cent VIJI) (per cent VIV)
Iofrared absorption spectrophotometry (2.2.24). 0-2 92 8
Comparison: DL-Iysine aCfJlylsalicylate CRS. 2 - 22 92 -+ 73 8 -+ 27
22 - 45 73 -+ 45 27 ..... 55
TESTS
Solution S
Dissolve 5.0 g in carbon dioxide-free water R and dilute '0 Flow rate 1.5 mUmin.
50 mL with the same solvent. Daecdon Spectrophotometer at 205 nm.
Appearance of solution Injection 10 ilL of the test solution and reference
Solution S is clear (2.2.1) and colourless (2.2.2, Me/hod II). solutions (b), (e), (I), (g) and (b).
pH (2.2.3) Identification of impun·ties Use the chromatogram obtained
4.5 to 6.0 for solution S. with reference solution (h) to identify the peak due £0
impurity Aj use the chromatogram obtained with reference
Related substances
solution (t) £0 identify Ute peaks due to impurities C and G;
use lite chromatogram obtainedwith reference solution (g) to
identify the peaksdue to impurities E and F.
Solvent mixture Mobilephase B, mobile phase A
Relativeretention With reference to acetylsalicylic acid
(10:90 VW).
(retention time = about 11 min): impurity G = about 0.2;
Solu.ion A Dilute 30 mL of a 103 gIL solution of impurity E = about 0.3; impurity F = about 0.4; DL-
hydrochlori< acidR to 1000 mL with the solvent mixture. lysine = about 0.9; impurity A = about 1.4;
Test solution Dissolve 0.100 g of the substance to be impurity C = about 1.7.
examined in 20 mL of solutionA and dilute to 25.0 mL with System suitability:
solution A. - resolution: minimum 3.5 between the peaks due to lysine
Reference solution (a) Dissolve 50 mg of aCfJlylsalicylic acid R and acetylsalicylic acid in the chromatogram obtained
in 2 mL of acetonitrile R and dilute to 50 mL with solution A. with reference solution (b);
Reference soluhon (b) Dissolve 0.100 g of lysine - signal-to-noise ratio: minimum 200 for the principal peak in
hydrochloride R in solution A, add 0.5 mL of reference the chromatogram obtainedwith reference solution (c).
solution (a) and dilute to 50 mL with solution A. Calculation of percentage contents:
Reference solution (c) Dilute 1.0 mL of the test solution '0 - correction factors: multiply the peak areas of the following
100.0 mL with solution A. Dilute 1.0 mL of this solution to impurities by the corresponding correction factor:
10.0 mL with solution A. = =
impurity C 4.3; impurity E 3.4; impurity F 4.3; =
Reference solution (d) Dissolve 2 mg of DL-lysine impurity G = 2.8;
acelylsalicylate impuniy C CRS in solution A and dilute to - for impurity A J use the concentration of salicylic acid in
50 mL with solution A. reference solution (h);
- for impurities other than A, use the concentration of DL-
Reference solut;"n (e) Dissolve 2 mg of DL-Iysine lysine acetylsalicylate in reference solution (c) taking into
acety/sa&y/au impun"ty G CRS in solutionA and dilute to account the area of the peak due to acetylsalicylic acid in
10 mL with solution A. reference solution (c).
Reference solution (f) To 3 mL of reference solution (d) add
Limits:
1 mL of reference solution (e) and dilute to 10 mL with
solutionA. - impurities E, F, G: for each impurity, maximum
Reference solut;"n (g) Dissolve 5 mg of N-(£)-acelyl-L-Iysine R 0.5 per .cent;
(I enantiomer of impurity E) and 5 mg of N-(.)-acelyl-L- - impun'ty C: maximum 0.3 per cent;
lysine R (1 enantiomer of impurity F) in solution A and - unspecified impun·lies: for each impurity, maximwn
dilute '0 10 mL with solution A. Dilute I mL of the solution 0.05 per cent;
to 20 mL with solution A. - ectal: maximum 2.0 per cent;
Reference solution (/I) Dissolve 20.0 mg of salicylic acid R - reporting threshold: 0.03 per cent; disregard the peak due to
(impurity A) in a mixture of equal volumes of acetonitrile R DL-Iysine.
and solution A and dilute to 5.0 mL with the same mixture Water (2.5.32)
of solvents. Dilute 1.0 mL of the solution to 100.0 mL with Maximum 0.3 per cent, determined on 0.300 g using the
solution A. evaporation technique at 100°C.
Co/umn: Sulfated ash (2.4.14)
- size: 1= 0.15 m, 12) = 4.6 nun; Maximum 0.1 per cent, determined on 1.0 g.
- stationary phase: end-capped amidohexadecylsilyl silica gelfor
chromatography R (5 um); ASSAY
- temperature: 30°C. Dissolve 0.140 gin 50.0 mL of anhydrous acetic acid R.
Titratewith o. J M perchknic add, determining the end-point
potentiometrically (2.2.21J).
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1-218 Aspirin Lysine 2022
1 mL of 0.1 M perchlotic acid is equivalent to 16.32 mg of o H CO~
CI5H22N206' ~~~NH2 aodenanuomer

IMPURITIES
Specified impurities A, C, E, F, G.
V OH
Other detectable impurities (the following substances would, if H. (2RS)-6-amino-2-(2-hydroxybenzamido)hexanoic acid,

present at a sufficient level, be deuced by oneor other of the tests
in themonograph. They arelimited by thegeneral acceptance OHC~H 0
~~~~J-..CH3
cnteiion for other/unspecified impurities and/or by thegeneral
manograph SubS/ances far pharmaceutical use (2034). II is end eoentcmer
therefore not necessary to idenuJy these jmpuniies for

demonstration of compliance. See also 5.10. Control of impun'ties
V OH
in substances for pharmaceutical use) B, D, H, l, JJ K, L, M.

I. (2RS)-6-ace13mido-2-(2-hydsoxybenzamido)hexanoic acid,
C0 2H
OC
"- OH ~~~~H
o H NHz
aodenanliomer
A. 2-hydroxybenzoic acid (salicylic acid),

U OH
J. (2RS)-2-amino-6-(2-hydsoxybenzamido)hexanoic acid,
o H~ H 0
.i.
eX:
-'1
"s epimet at C' and theirenanUomers
:::? I ~~~ ~ andeoanUomer
"- OH
B. (2RS)-6-amino-2-[(2R)-2,6-diaminohexanamido)hexanoic
acid and (2RS)-6-amino-2-[(2S)-2,6-diaminohexanamido) K. (2RS)-2-ace13mido-6-(2-hydroxybenzamido)hexanoic acid,
hexanoic acid,
H NH2 H
~ »<: X ~N~CO,H
H~ " " l(' ;\
o H N~
itseplmer at C' and theirenantlomers
C. (2RS)-2-amino-6-[(2R)-2,6-diaminohexanamido]hexanoic L 2-[[2-(acetyloxy)benzoyl)oxy)benzoic acid

acid and (2RS)-2-amino-6-[(2S)-2,6-diaminohexanamido] (acetylsalicylsalicylic acid),
hexanoic acid,
(XI Co,H
O~
andenanllomer
D. (3RS)-3-aminoazepan-2-one,
M.2-[(2-hydsoxybenzoyl)oxy)benzoic acid (salselate,
salicylsalicylic acid).
and ereouome- _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII
E. (2RS)-6-acelamido-2-aminohexanoic acid (N-(E)-acetyl-

DL-lysine),
F. (2RS)-2-acelamido-6-aminohexanoic acid (N-(a)-acetyl-

DL-Jysine),
o H~C H 0
)l - \J)l and enan_
~c ~~~ c~
G. (2RS)-2,6-diacetamidohexanoic acid,
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2022 Atazanavir Sulfate 1-219
Atazanavir Sulfate *** Reference solution (b) Dilute 1.0 mL of test solution (a) to
*** *** 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
(ph. Eur. monograph 2898) *** solution to 10.0 mL with me solvent mixture.
Reference solution (c) Dissolve 4 mg of alazanav;rfor system
suitability CRS (containing impurity F) in 8 rnL of the solvent
mixture, sonicate for 3 min and dilute to 10 mL with the
solvent mixture.
Reference solution (d) Dissolve 2.0 mg of atazanaoir
impun'ty K CRS in 9 mL of the solvent mixture, sonicate for
3 min and dilute to 10.0 mL with the solvent mixture. Dilute
5.0 mL of the solution to 100.0 mL with the solvent mixture.
Dilute 3.0 mL of this solution to 20.0 mL with the solvent
mixture.
Column:
=
- size: 1 0.15 01, 0 4.6 mm; =
803 229975-97-7 - staiionary phase: end-copped octadeeylsilyl silica gelfor
chromawgraphy R (3.0 ~m);
Action and use - temperature: 25°C.
Antiviral (HIV). Mobile phase:
PhE" _
- mobile phase A: mix 25 volumes of acetonitrile Rl and
75 volumes of a freshly prepared 2.73 gIL solution of
DEFINITION potassium dihydrogen phosphate R previously adjusted to
Methyl [(5S, lOS, I IS,14S)-1l-benzyl-5-Wl-butyl-lO-hydroxy- pH 3.5 with dilute phosphoric acid R;
I 5, 15-dimethyl-3,6, 13-trioxo-8-[[4- (pyridin-2-yl)phenyl] - mobile phase B: mix 25 volumes of a freshly prepared
methyJ]-2-oxa-4,7,8, 12-tetraazahexadecan-14-yl]carbamate 2.73 gIL solution of potassium dihydrogen phosphate R
sulfate. previously adjusted to pH 3.5 with dilute phosphoric acid R,
and 75 volumes of acetonitrile Rl;
Content
Time Mobile phase A MobUe phase B
CHARACTERS (mID) (per eenr VIJI) (per cent V/J?
Appearance 0-' 100 0
White or pale yellow, slightly hygroscopic, crystalline powder 5 - 45 100 ..... 0 0 ..... 100
that may contain agglomerates.
Solubility Flow rate 1.0 mUmin.
Slightly soluble in water, freely soluhle in ethanol Detection Spectrophotometer at 215 nm.
(96 per cent), practically insoluble in heptane. Injection 10 J.1L of test solution (a) and reference
IDENTIFICATION solutions (b) and (c).
A. Specific optical rotation (see Tests). Identification of impuriti<s Use the chromatogram supplied
B. Infrared absorption spectrophotometry (2.2.24). with ataeonodr for system suitability CRS and the
Comparison atazanavir sulfateCRS. chromatogram obtained with reference solution (c) to identify
the peak due to impurity F.
C. It gives reaction (a) of sulfates (2.3.1).
Re/alifJe retention With reference to atazanavir (retention
TESTS =
time about 30 min): impurity F about 0.99. =
-44 to -40 (anhydrous substance), measured at 25 "C.
Dissolve 0.100 g in 8 mL of methanol R, using sonication if impurity F and atazanavir.
necessary, and dilute to 10.0 mL with the same solvent. Calculation of percentage contents:
Related substances - for each impurity, use the concentration of atazanavir
Liquid chromatography (2.2.29). sulfate in reference solution (b).
Solvent mixture Mix equal volumes of acetonitrile Rl and a Limits:
freshly prepared 2.73 gIL solution of potassium dihydrogen - unspecified impurities: for each impurity, maximum
phosphate R in waterfor chromawgraphy R previously adjusted 0.10 per cent;
to pH 3.5 with dilute phosphoric acid R. - total: maximum 0.5 per cent;
Test solution (a) Dissolve 20.0 mg of the substance to be - reporting threshold: 0.05 per cent; disregard any peak with a
examined in 40 mL of the solvent mixture, sonicate for relative retention with reference to atazanavir of less than
3 min and dilute to 50.0 mL with the solvent mixture. 0.2.
Test solution (b) Dissolve 50.0 mg of the substance to be ImpurityK
examined in 40 mL of the solvent mixture, sonicate for Liquid chromatography (2.2.29) as described in the test for
3 min and dilute to 50.0 mL with the solvent mixture. related substances with the following modifications.
Reference solution (a) Dissolve 20.0 mg of ataztmauir MoMe phase:
sulfate CRS in 40 mL of the solvent mixture, sonicate for
3 min and dilute to 50.0 mL with the solvent mixture.
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1-220 Atazanavir Sulfate 2022
Tim. MobUe phase A MobUe phase B

(min) (per cent VIII) (per cent VIJI)
o-S 9S S
S- 8 95 ---> 0 5 -0100
8 - 14 0 100
B. 4-(pyridin-2-yl)benzaldehyde,
Injection 20 ilL of [est solution (b) and reference
solution (d).
ldentification of impunties Use the chromatogram obtained
with reference solution (d) to identify the peak due to
impurity K.
Relative retention With reference to atazanavir (retention
time =about 10 min): impurity K =about 0.4.
Calculation of percentage content:
- for impurity K, use the concentration of impurity K in
reference solution (d).
C. methyl [(5S,IOS,IIS,14S)-II-benzyl-5-tert-butyl-lO-
Limit: hydroxy-l 5, 15-dimethyl-3,6, 13-trioxo-2-oxa-4,1,8,12-
- impun"ty K: maximum 0.15 per cent. letraazahexadecan-14-yl]carbamate,
Water (2.5.32)
Maximum 2.5 per cent, determined on 0.100 g by direct
sampleintroduction.
ASSAY
Mobile phase Solvent mixture.
Injection Test solution (a) and reference solutions (a) and
(c).
D. (2S,3S)-3-amino-4-phenyl-l-[(8) -1- [[4-(pyridin-2-yl)
Run time 1.6 times the retention time of atazanavir, phenyl]methyl]-2-[[4-(pyridin-2-yl)phenyl]methylidene]
Relative retention With reference to atazanavir (retention hydrazin-I-yl]butan-2-01,
time =about 9.5 min): impurity F =about 0.94.
impurity F and atazanavir.
Calculate the percentage content of C38Hj~6011S using the
chromatogram obtainedwith reference solution (a) and
taking into account the assigned content of atazanamr
sulfate CRS.
STORAGE
IMPURITIES
Specified impurities K. E. methyl [(5S,IOR,IIS,14S)-II-benzyl-5-'ert-butyl-lO-
Other detectable impurities (the following substances would, if hydroxy-I 5, 15-dimethyl-3,6, 13-rrioxo-8-[[4-(pyri din- 2-yl)
present at a sufficient level, be detected by oneor other of the tests phenyl]methyl]-2-oxa-4,1,8,12-tetraazahexadecan-14-yl]
in the monograph. They arelimited by the general acaptan« carbamate, .
monograph Substances for phannaceutical use (2034). 1, is
therefore not necessary to identify these impun"ties for
demonstration ofcompliance, See also 5.10. controlof impuri'ies
in substances for pharmaceutical use) A, B, 0, D, E, F, G, H,
1, J.
A. 4-(pyridin-2-yl)benzoic acid, F. methyl [(5R, lOS, liS, 14S)-II-benzyl-5-tert-butyl-lO-

hydroxy-l 5, 15-dimethyl-3,6,13-trioxo-8-[[4-(pyridin-2-yl)
phenyl]methyl]-2-oxa-4,1,8,12-tetraazahexadecan-14-yl]
carbamate,
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2022 Atenolol 1-221
K. (2S)-2-(methoxyfonnamido)-3,3-dimethylbutanoic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'I>E<r
Atenolol ***
G. methyl [(5S, lOS, I IS,14R)-1l-benzyl-5-tert-butyl-lO- *** *
*
hydroxy-l 5, 15-dimethyl-3,6, 13-trioxo-8-[[4-(pyridin-2-yl) (ph. Eur. monograph 0703) ****
phenyl]methyl]-2-oxa-4,7,8,12-tetraazahexadecan-14-yl]
carbamate, H pH H
JJ)
0~ N
o I" Y CH, and enanllomer
.# CH3
H,N
266.3 29122-68-7
Action and use

Beta-adrenoceptor antagonist.
Preparadons
Atenalol Injection
Atenolcl Oral Solution
AtenalolTablets
H. methyl [(5S, lOR, IIR, 14S)-II-benzyl-5-tert-butyl-10-
Co-tenidone Tablets
hydroxy-I 5, 15-dimethyl-3,6,13-trioxo-8-[[4-(pyridin-2-yl)
phenyl) methyl]-2-oxa-4,7,8, 12-tetraazahexadecan-14-yl] I'I>E<r ~
carbamate,
DEFINITION
2-[4-[(2RS)-2-Hydroxy-3-[(propan-2-yl)amino]
~
N
I propoxy]phenyl] acetamide.
""""'" ° Content
uU ,
'"" H
ON "N'
1;
1.J...H_OCH'
' HH.NI(.'X_CH3
'" 0 H,C CH,
CHARACTERS
Appearance
While or almost white powder.
Solubility
Sparingly soluble in water, soluble in anhydrous ethanol,
slightly soluble in methylene chloride.
I. methyl [(2S)-1-[[(2S,3S)-3-hydroxy-I-phenyl-4-[(E)-I-
IDENTIFICATION
[[4-(pyridin-2-yl)phenyl] methyl]-2-[[4-(pyridin-2-yl)
phenyl] methylidene]hydrazin-I-yl] butan-2-yl) amino]-3,3-
dimethyl-Loxobutan-2-yl]carbamateJ Second identification: A, C.
A. Melting point (2.2.14): 152 -c to 155 'C.
,r N B. Infrared absorption spectrophotometry (2.2.24).
""I Comparison atenolo! CRS.
I""'" C. Thin-layer chromatography (2.2.27).
H3C CH3 0"" HO... H H

examined in 1.0 mL of methanol R.
X :Jl
H,CO~ 'N1;N
\ y o -..".--CH
/\ ' Reference solutUm Dissolve 10 mg of atenolol CRS in 1.0 mL
H o~c CH3 of methanol R.
Plat< TLC silanised silica gel Fm plate R.
I,. Mobile phase concentrated ammonia Rl, methanol R
(1:99 VIII).
J. lert-butyl 2-[(2S,3S)-3-(tert-butoxyfonnamido)-2-hydroxy- Application I0 ~L.
4-phenylbutyl]-2-[[4-(pyridin-2-yl)phenyl]methyl] Development Over 3/4 of the plate.
hydrazine-l-carboxylare,
Dry;ng In air.
Detection Examine in ultraviolet light at 254 nm.
with the test solutionis similar in position and size to the
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1-222 Atenolol 2022
principal spot in the chromatogram obtained with the - total: not more than 5 times the areaof the principal peak
reference solution. in the chromatogram obtainedwith reference solution (b)
TESTS (0.5 per cent);
Solution S
the chromatogram obtainedwith reference solution (b)
(0.05 per cent).
same solvent.
Chlorides (2.4.4)
Solution S is dear (2.2.1) and not more intensely coloured
than intensity 6 of the range of reference solutions of the Dissolve 50 mg in a mixture of 1 mL of dilute nittic acid R
most appropriate colour (2.2.2, Method II). and 15 mL of Water R. The solution, without further addition
of dilute nitric acid R, complies with the test.
-0.10° to + 0.10°, determined on solution S. Loss on drying (2.2.32)
Related substances
an oven at 105 "C.
Testsolution Dissolve 50 mg of me substance to be
examined in 20 mL of the mobilephase and dilute to
25.0 mL with the mobile phase. ASSAY
Reference solution (aJ Dissolve 2 mg of arenolol for sysum Dissolve 0.200 gin 80 mL of anhydrous acetic acid R. Titrate
sln"lability CRS (containing impurities B, F, G, I and D in with 0.1 M perchlon·c acid, determining the end-point
1 mL of the mobile phase. potentiometrically (2.2.20).
Reference sdution (b) Dilute 1.0 mL of the test solution to 1 mL of 0.1 M perchloric acid is equivalent to 26.63 mg of
100.0 mL with the mobile phase. Dilute 1.0 mL of this C,.,H"N2 0 , .
solution to 10.0 mL with the mobile phase. IMPURITIES
Column: Specified impurities B, F, G, 1, J.
-size: 1= 0.125 m, Q) = 4.0 mm; Other detectable impurities (the following substances would, if
- stationary phase: end-capped octad«ylsilyl silira gelfor present at a sufficient level, be detected by one or other of the tests
chromatography R (5 um). in the monogroph. They aro limited Iry the generol acceptance
Mobile phase Dissolve 1.0 g of sodium octanesulfonate Rand cntetion for otherlunspe<ified impuritks and/orIry thegeneral
0.4 g of urraburylammoniu,m hydrogen sulfa" R in 1000 mL of monograph Substances for phatmaceuti<al use (2034). It is
a mixtureof 20 volumes of tetrahydrofuran R, 180 volumes of therefore not necessary to identify these impurities for
methanol R2 and 800 volumes of a 3.4 gIL solution of demonstration of compliance. See abo 5.10. Control of impurities
potassium dihydrogen phospha" R; adjust the apparent pH to in substances for pharmaceutical use) A, D, E, H.
3.0 with phosphoric acidR.
Flow rate 0.6 mllrnin.
Injection 10 ~L.
Run time 5 times the retention time of atenolol.
A.2-(4-hydroxyphenyl)acetamide,
Identification of impun'ties Use the chromatogram supplied
with a"nololfor syuem suitability CRS and the chromatogram H OH
obtained with reference solution (a) to identify the peaks due '2 IfYO~OH
to impurities B, F, G, I and J.
~ andenanliomer
Relativeretention With reference to atenolol (retention H,N
time e about 8 min): impurity B ;::; about O.3j
impurity J = about 0.7j impurity I = about 0.8; B. 2-[4-[(2RS)-2,3-dihydroxypropoxy]phenyl]acetantide,
impurity F = about 2.0 (pair of peaks);
=
impurity G about 3.5. . H OH
System su£tability Reference solution (a): '2 0r0~CI
- resolution: minimum 1.4 between the peaks due to ~ andenantiomer
impurities J and I. H,N
Limits:
- impwity B: not more than twice the area of the principal D.2-[4-[(2RS)-3-cWoro-2-hydroxypropoxy]phenyl]acetamide,
solution (h) (0.2 per cent);
- impurities F, G, 1, J: for each impurity, not more than
(0.15 per cent);
- unspecified impurities: for each impurity, not more than the E. 2,2'-[(2-hydroxypropane-I,3-diyl) bis(oxy-4, I-phenylene)]
area of the principal peak in the chromatogram obtained dlacetamide,
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2022 Atomoxetine Hydrochloride 1-223
H3C
yCH3 Solubility
OH I OH Sparingly soluble in water, soluble in anhydrous ethanol,
,10 0~ N ~ODJL practically insoluble in heptane.

o I I 0
H:zN # .,# NH2
IDENTIFICATION
F. 2,2'-[[(propan-2-yl)azanediylj bis[(2-hydroxypropane- 3, 1- A. Infrared absorption spectrophotometry (2.2.24).
diyl)oxy-4,I-phenylene]jdiacetamide, Comparison oiomoxedne hydrochloride CRS.
»
H pH H
v dissolve the substance to he examined and the reference
~
0
~
N
y CH,
andenantlomer substance separately in anhydrous ethanol R, evaporate (Q
H0:2C # CH3 dryness and record new spectra using the residues.
B. Isomeric purity (see Tests).
G. [4-[(2RS}-2-hydroxy-3-[(propan-2-yl)amino]propOXY] C. It gives reaction (a) of chlorides (2.3.1).
phenyl]acetic acid,
TESTS
H OH
»
Isomeric purity
~
I ~
0
~
V
y CH,
andenantiomer Liquid chromatography (2.2.29): use the norrnalisation
procedure.
Ne.-? CH3
H. [4-[(2RS}-2-hydroxy-3-[(propan-2-yl)amino]propoxyj examined in 2.5 mL of anhydrous ethanol R, sonicate until
phenyl]acetonitrile, dissolution is complete and dilute to 10.0 mL with heptane R.
Reference solution (a) Dissolve 3.5 mg of atomoxeeme
impurity B CRS and I mg of awmaxetine impurity D CRS in
5 mL of anhydrous ethanol R, sonicate until dissolution is
complete and dilute to 20.0 mL with heptane R.
Reference solution (1)) Dissolve 35.0 mg of the substance to
be examined in 2.5 mL of anhydrous ethanol R. Add 1.0 mL
I. 2-[4-[(2RS}-3-(ethylamino)-2-hydroxypropoxyjphenylj of reference solution (a) and dilute to 10.0 mL with
acetamide, heptane R.
H OH Reference solution (c) Dilute 1.0 mL of reference solution (a)
,10
0 ~ NH2 to 100.0 mL with heptane R.
I
o and enanliomer Column:
H,N D' - size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: cellulose denvative of silica gelfor chiral
J. 2-[4-[(2RS}-3-amino-2-hydroxypropoxyjphenyl]acetamide. separation R (5 pm).
____ ~_~~~ ~ PIIE"' Mobile phase Mix 1.5 mL of diethylamine R, 2.0 mL of
trifluoroacetic acid R and 150.0 mL of 2-propanol R and dilute
to 1000 mL with heptane R.
Atomoxetine Hydrochloride Detection Spectrophotometer at 273 nm.
Injection 10 J1L of the test solution and reference
(Ph. Eur. monograph 2640) solutions (b) and (c).
Run lime 1.3 times the retention time of atomoxetine.
Identification of impuniiu Use the chromatogram obtained
, Hel with reference solution (b) to identify the peaks due to
impurities Band D.
Rekuiue retention With reference to atomoxetine (retention
time = about 12 min): impurity B = about0.5;
291.8 82241J.59-7 impurity D = about 0.6.
Action and use
Noradrenaline reuptake inhibitor, treatment of attention
impurities Band D.
deficit hyperactivity disorder (ADHD).
Limits:
PIlE", _
~
- impun'ty B: maximum 0.5 per cent;

DEFINITION - impun"ty D: maximum 0.15 per cent;
(3R)-N-Methyl-3-(2-methylphenoxy)-3-phenylpropan-l- - unspecified impurities: for each impurity, maximum
0.10 per cent;
amine hydrochloride.
- disregard Hmit: the area of the peak due to impurity B in
Content the chromatogram obtained with reference solution (c)
98.0 per cent to 102.0 per cent (dried substance). (0.05 per cent); disregard any peakwith a relative
CHARACTERS retention with reference to atomoxetine of about 0.7
Appearance (impurity A).
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1-224 Atomoxetine Hydrochloride 2022
Related substances ASSAY

Liquid chromatography (2.2.29). Liquid chromatography (2.2.29) as described in the test for
Solution A Dissolve 5.9 g of sodium ocomesulfonau related substances with the following modification.
monohydrate R in 1000 mL of a 2.9 gIL solution of phosphom Injection Test solution (b) and reference solution (d).
acidR previously adjusted to pH 2.5 with a 280 gIL solution Calculate the percentage content of C I7H22CINO taking into
of potassium hydroxide R. account the assigned content of atomoxetine
Test solution (aJ Dissolve 25 mg of the substance to be hydrochloride CRS.
IMPURITIES
the mobilephase.
Specified impurities A, B, D.
Test solution (b) Dissolve 25.0 mg of the substance to be
Other detecrable impurities (,he following substances would, .l
present at a sufficien, level, be detected by oneor other of the tests
the mobilephase.
in the monograph. They arelimited by thegeneral acceptance
Reference solu,wn (a) Dilute 1.0 mL of test solution (a) to ctitenon for other/unspecified impun"ties andior by lire general
100.0 mL with the mobile phase. Dilute 1.0 mL of this monograph Substances for pharmaceutical use (2034). 1, is
solution to 10.0 mL with the mobile phase. therefore not necessary to identify these impurities for
Refemce sonuion (b) Dissolve 7.5 mg of 3-(methylamino)-I- demonstration of compliance. See olso 5.10. Control of impurities
phenyipropan-l-ol R (impurity H) and 5 mg of mandelic acidR in substances for pharmaceutical use) G, E, F, G, H.
(impurity E) in test solution (b) and dilute to 50 mL with
test solution (b).
Reference solution (e) Dissolve 5 mg of atomoxetine lor
impurify A identification CRS in the mobile phase and dilute
to 20 mL with the mobile phase.
Reference solution (d) Dissolve 25.0 mg of aomoxedne
hydrochloride CRS in the mobile phase and dilute to
A. N-methyl-3-phenoxy-3-phenylpropan-l-amine,
Column:
- size: 1= 0.15 m, 0 = 4.6 mmj
- sra,ionary phase: end-eapped oay/silylsitka gelfor
chromatography R (3.5 urn);
Mobile phase propanol R, solution A (27:73 VIl').
Flow role 1.0 mUmin. B. (3S)-N-methyl-3-(2-methylphenoxy)-3-phenyJpropan-l-
Detection Spectrophotometer at 215 nm. amine,
Injectiim 10 Jll. of test solution (a) and reference
Run time 2.5 times the retention time of atomoxetine,
Identification of impun"ties Use the chromatogram obtained
with reference solution (b) to identify the peaks due to
impurities E and H; use the chromatogram supplied with C. (3R)-N-methyl-3-(4-methylphenoxy)-3-phenylpropan-l-
atomoxetine for impurity A identifirouon CRS and the amine,
chromatogram obtained with reference solution(c) to identify
the peak due to impurity A.
Relative retention With reference to atomoxetine (retention
time ~ about 10 min): impurity E ~ about 0.2;
impurity H ~ about 0.3; impurity A ~ about 0.7.
SYSW1l suitability Reference solution (b):
- resolution: minimum 5.0 between the peaks due to D. (3R)-N-methyl-3-(3-methylphenoxy)-3-phenylpropan-l-
impurities E and H. amine,
HO H
- for each impurity, use the concentration of atornoxetine
hydrochloride in reference solution (a).
Limits:
- impun'ty A: maximum 0.3 per cent;
o>
E. (2S)-2-hydsoxy-2-phenylacetic acid (t-mandelic acid),
0.10 per cent;
- UJtaI: maximum 0.5 per cent;
Maximum 0.5 per cent, determinedon 1.000 g by drying in
vacuo at 105°C for 2 h.
Maximum 0.1 per cent, determinedon 1.0 g- F. (3S)-3-(3-fluoro-2-methylphenoxy)-N-methyl-3-
phenylpropan-l-amine,
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2022 Atorvastatin Calcium Trihydrate 1-225
TESTS
Enantiomeric purity
Solvent mixture anhydrous ethanol R, methanol R
(50:50 VII').
G. 3,3'-[(2-methylbenzene-I,3-diyl)bis(oxy)]bis(N-methyl-3- examined in 4 mL of the solvent mixture and dilute to
phenylpropan-f-amine), 10.0 mL with hexone R.
Reference solution (a) Dissolve 2 mg of atorvastatin
impurityE CRS in methonol R and dilute to 20.0 mL with the
same solvent (solution A). Dissolve 10 mg of the substance
to be examined in 1.25 mL of methonol R, add 0.75 mL of
solution A and 2 mL of anhydrous ethanol R and dilute to
10.0 mL with hexone R.
H. 3-(methylamino)-I-phenylpropan-I-01. Reference solution (b) To 2.0 mL of the test solution add
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POE.. 40.0 mL of the solventmixture and dilute to 100.0 mL with
hexone R. To 3.0 mL of this solution add 5 mL of the
solvent mixture and dilute to 20.0 mL with hexane R.
Column:
Atorvastatin Calcium Trihydrate - size: 1;;;; 0.25 m, 0 = 4.6 mm;
- stationary phase: amylose den·vative of silica gelfor
(Ph. Bur. monograph 2191) chromatography R (10 urn).
Mobile phase trijluoroacetit acid R, anhydrous ethanol R,
CH3 c~-
hexone R (0.1:6:94 VIVII').
N
~OH
H
Flow ro,. 1.0 mllmin.
ca" , 3H zO
Injection 20 ~L.
Run time 1.2 times the retention time of atorvastatin.
Relative retention With reference to atcrvastatin (retention
F
= =
time about 44 min): impurity E about 0.8.
344423-98-9 - resolution: minimum 2.0 between the peaks due to
impurity E and atorvastatin.
Action and use
HMG Co-A reductase inhibitor; lipid-regulating drug. Limit:
- impurity E: not more than the area of the principal peak in
POE.. _ the chromatogram obtained with reference solution (b)
(0.3 per cent).
DEFINITION
Calcium (3R,5R)-7- [2-(4-f1uorophenyl)-5-(I-methyl ethyl)-3- Related substances
phenyl-4-(phenylcarbamoyl)-IH-pyrrol-I-yl)-3,5- Liquid chromatography (2.2.29).
dihydroxyheptanoate trihydrate. Test solution (a) Dissolve 40.0 mg of the substance to be
Content examined in dimethylformamide R and dilute to 100.0 mL
97.0 per cent to 102.0 per cent (anhydrous substance). with the same solvent.
Test solution (b) Dissolve 50 mg of the substance to be
CHARACTERS examined in dimethylfonnamide R and dilute to 50.0 mL with
Appearance the same solvent.
Reference solution (a) Dissolve 40.0 mg of atorvastatin
Solubility calcium t,,·hydrate CRS in dimethylformamiJe R and dilute to
Very slightly soluble in water, slightly soluble in ethanol 100.0 mL with the same solvent.
Reference solution (b) Dilute 1.0 mL of test solution (b) to
II shows polymorphism (5.9). 100.0 mL with dimethylformamide R. Dilute 1.0 mL of this
IDENTIFICATION solution to 10.0 mL with dimethylfomlOmide R.
A. Infrared absorption spectrophotometry (2.2.24). Reference solution (c) Dissolve 2 mg of atoroastatin for system
Comparison ateroQSeatin calcium trihydrate eRSt suitability CRS (containing impurities A, B, C and D) in
If the spectra obtained in me solid stateshow differences, dimethylfonnamide R and dilute to 5 mL with the same
dissolve the substance to he examined and the reference solvent.
substance separately in methanol R, evaporate [Q dryness and Column:
record new spectra using the residues. - size: I = 0.25 m, 0 ;;;; 4.6 mm;
B. Enantiomeric purity (see Tests). - stationary phase: oaylsilylsilica gelfor chromatography R
(5 um);
C. Water (see Tests). - temperature: 35 "C.
D. Ignite. The residue gives reaction (b) of calcium (2.3.1).
Filtration may he necessary in case the residue does not
completely dissolve.
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1-226 Atorvastatin Calcium Trihydrate 2022
Mobile phase: SQurce Sodium hollow-cathode lamp.

- mobile phase A: tetrahydrofuron R, aceionimle R, 3.9 gIL Wavelength 589.0 om.
solution of ammonium acetate R adjusted to pH 5.0 with
Awmisation device Air-acetylene flame.
glacial acetic acid R (12:21:61 VIVIV);
- mobae phaseB: tetrahydrofuran R, 3.9 gIL solution of Water (2.5.12)
ammonium acetate R adjusted to pH 5.0 with glcu:"a/ acetic 3.5 per cent to 5.5 per cent, determined on 0.130 g.
acid R, acetonitrile R (12:21:61 VIVIV); ASSAY
Tim. Moblle phase A MobUe phase B related substances with the following modification.
(min) (per cent VIP) (per cent VIII)
InjC€ti~ Test solution (a) and reference solution (a).
0-40 100 0
4.0·70 100 -> 20 0->80
Calculate the percentage content of C6c)168CaF2N"OlO from
70 - 85 20 -+ 0 80 -> 100
me declared content of atoruastatin calcium trihydrate CRS.
IMPURITlliS
Flow rate 1.5 mUmin. Specified impuruies A, B, C, D, E.
Detection Spectrophotometer at 244 nm. Other dete</able impun·'" (the following substances would, if
Injection 20 IJL of test solution (b) and reference present at a sufficient level, be detected by ~e or other of the tests
solutions (b) and (c). in the monograph. They are limited by thegeneral a«epUlnce
cnierion for otherlumpecified impun't'" andlor by thegenerol
Identification of impuriues Use the chromatogram obtained
monograph Subsrances for pharmaceutical use (2034). II is
with reference solution (c) to identify the peaks due to
therefore not necasary to identify these impurities for
impurities A, B, C and D.
demonstration of compliance. See also 5.10. Control of impuniies
Relative retention With reference to atorvastatin (retention in substances for pharmaceutical use) P, G, H.
time =about 33 min): impurity A =about 0.8;
impurity B ;;;; about 0.9; impurity C ;;;; about 1.2; li,C CIi, Co,H
If necessary, adjust the mobile phase by increasing or
decreasing the percentage of acetonitrile or the pH of the
O
I '"
.-P
~
0
.p'
N
~-OH
H
H-. pH (
ammoniwn acetate solution to achieve a retention time of

about 33 min for atorvastatin, For example, raising the pH
would decrease the retention time of atorvastatin.
Sysrem suitability Reference solution (c):
A. (3R,5R) -3,5-<1ihydroxy-1-[5-( I-methylethyl)-2,3-diphenyl-
- peak-w-tJa//ey ratio: minimum 1.5, where Hp = height
4-(phenylcarbamoyl)-IH-pyrrol-l-yl] heptanoic acid
(desfiuoroatorvastarin),
HlJ = heightabove the baselineof the lowest point of the
curve separating thispeak from the peak due to
atorvastatin,
('1
Limits: ~NH
- impuniies A, B: for each impurity, not more than 3 times
obtainedwith reference solution (b) (0.3 per cent);
andenantiomer
- impuniies C, D: for each impurity, not more than
1.5 times the area of the principal peak in the F
(0.15 per cent); B. (3RS,5SR)-1-[2-( 4-fluorophenyl)-5-(l-methylethyl)-3-
- unspecified impurities: for each impurity, not more than the phenyl-4-(phenylcarbamoyl)-1 H-pyrrol-I-yl] -3,5-
area of the principal peak in the chromatogram obtained dihydroxyheptanoic acid,
(0.05 per cent); disregard the peak due to
dimethylformamide.
Sodium F F
Maximum 0.4 per cent (anhydrous substance).
Atomic absorption spectrometry (2.2.23, Method I).
c. (3R,5R)-1-[2,3-bis(4-fluorophenyl)-5-( I-methylethyl) -4-
(phenylcarbamoyl)-I H-pyrrol-I-yl]-3,5-
Solventmixture hydrochloric add R, waterR, methanol R dihydroxyheptanoic acid (fluoroatorvastatin),
(2:25:15 VIVIV).
Testsolution Dissolve 5.0 mg in the solvent mixture and
dilute to 100.0 mL with the solventmixture.
Reference solutions Prepare the reference solutions using
sodium standardsolution (50 ppm Na) R, diluting with the
solvent mixture.
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2022 Atovaquone 1-227
H,C CH,
Atovaquone ***
Q N
H
0
0
*** ***
***
P
F
«X
'" -::p"
0 H~' H::?"
•• "
I I I
D.3-[(4-t1uorophenyl)carbonyl]-2-(2-methylpropanoyl)-N,3- "'::;,.. OH 'C1
diphenyloxirane-2-carboxamide, o
C"H 19CIO, 366.8 95233-18-4
Action and use

Antiprotozoal (malaria).
'''''" - - - - - - - - - - - - - - - - - - -
DEFINITION
F
2-[trans-4-(4-CWorophenyl)cyclohexyl]-3-
E. (3S,5S) -7- [2-(4-t1uorophenyl) -5-( l-methylethyl)-3-phenyl- hydroxynaphthalene-I,4-dione.
4-(phenylcarbamoyl)-IH-pyrrol-l-yl]-3,5- Content
dihydroxyheptanoic acid (ern-atorvastarln), 97.5 per cent to 102.0 per cent (anhydrous substance).
HOHHOH
CHARACTERS
Ho,C~NH Appearance
Yellow, crystalline powder.
oH,C C~'
H OH 0 Solubility
, • -'OH Practically insoluble in water, sparingly soluble in methylene
N H chloride, very slightly soluble in methanol.
IDENTIFICATION
F Comparison alOVaquone CRS.
If the spectra obtained show differences, dissolve 0.1 g of the
F. (3R,5R)-7-[[(3R,5R)-7 -[2-(4-t1uorophenyl)-5-(I-
substance to be examined and 0.1 g of the reference
methylethyl)-3-phenyl-4-(phenylcarbamoyl)-IH-pyrrol-l-
substance separately in 2.5 mL of a 50 gIL solution of
yl]-3,5-dihydroxyheptanoyl]arnino]-3,5-
potassium hydroxide R in methanolR. Filter the solutions and
dihydroxyheptanoic acid,
add each filtrate dropwise to a mixture of 0.8 mL of acetic
acidRand 1.5 mL of methanol R, stirring continuously.
Filter, wash the residues with methanol R and then with
water R, and dry under vacuum at 55°C. Record new
spectra using the residues.
TESTS
Related substances
Liquid chromatography (2.2.29). Cany out the USI protected
F .from lighl.
Solvenl mixture warer R, acetonirrile Rl (20:80 VIV).
G. (3R,5R)-7 -[2-(4-t1uorophenyl)-5-( l-methylethyl)-3-phenyl-
4-(phenylcarbarnoy1)-IH-pyrrol-I-yl] -5-hydroxy-3-
methoxyheptanoic acid (3-G-methylatQ[vastatin),
o Reference sdution (a) Dissolve 25.0 mg of alOVaquone CRS
oH,C CH~
0
in the solvent mixture and dilute to 100.0 mL with the
solvent mixture.
? "OH
N ~ H
Reference solution (b) Dissolve 2.5 mg of alOVaquom for
sysrem suilability CRS (containing imputities B and C) in the
solvent mixture and dilute [Q 10.0 rnL with the solvent
mixture.
F
H. (4R,6R) -6- [2- [2-(4- t1uorophenyl)-5-( I-methylethyl)-3-
phenyl-4-(phenylcarbamoyl)-IH-pyrrol-l-yl]ethyl]-4- Column:
hydroxyretrahydro-2H-pyran-2-one.
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped <xtadecylsilYl silica gelfor
_________________ ~ Ph'" chromatography R (5 pm).
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1-228 Atracurium Besilate 2022
AIobiJe phase phosphoric acid R, methanol R2, waterfor

chromawgraphy R, acetonitrile R1 (0.5:17.5:30:52.5 VIVIVIJI).
Deuaion Spectrophotometer at 220 run.
Injection 20 j.lLof the test solution and reference A. 2-[trans-4-(4-chlorophenyl)cyclohexyl]-I-oxo-1 H-indene-
solutions (b) and (c). 3-carboxylic acid,
Run time Twice the retention time of atovaquone,
~.~.~
Identification of impurities Use me chromatogram supplied
with alOfJaquone for system suilability GRS and the
identify the peaks due to impurities Band C. VY'OH ~CI
Relative retention Wlth reference to atovaquone (retention
o
time = about 15 min): impurity B = about 0.85; B. 2-[ cis-4-(4-chlorophenyl)cyclohexyl)- 3-
=
impurity C about 0.90. hydroxynaphthalene-l,4-dione,
impurity C and atovaquone; Cl
- peak-to-fJolky ratio: minimum 1.5, where Hp :;;: height
above the baseline of the peak due to impurity C and andenanliomer
=
H v height above the baseline of the lowest point of the o
impurity B. C. 2-[( IRS) -4-(4-chlorophenyl)cyclohex-3-en-l-yl)-3-
Calculation of percentage contents: hydroxynephrhatene-Ld-dione,
- for each impurity, use the concentration of atovaquone in o
W"%"
reference solution (c)"
Limits: ""I ( ""I
- impun"ty B: maximum 0.5 per cent; OCH3 CI
- impun'ty C: maximum 0.2 per cent; o
- unspecified impun"ties: for each impurity, maximum
0.10 per cent; . D.2-[nuns-4-(4-chlorophenyl)cyclohexyl)-3-
- total: maximum 0"6 per cent; methoxynaphthalene-l,4-dione.
- reporting threshold: 0.05 per cent. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf<6
Water (2.5.32)
Maximum 0.3 per cent, determined on 0.100 g using the
evaporation technique:
- temperature: 160 "C;
- heating time: 3 min;
Atracurium Besilate
- flow ral<: 50 mUmin. (ph. Eur. monograph 1970)
ASSAY
Calculate the percentage content OfC2JIl9C103 laking into
account the assigned content of awvaquone CRS.
IMPURITIES
Specified impurities B, G.
Otherdetulable impurities (the following subslances would, if
present at a sufficient level, be deteaed by oneor other of the tests
in the monograph. They are limited by the general a«tj>lance
cruetion for other/unspecified impurities and/or by the general
monograph SubSlances for pharmaceutical use (2034). It is 1243 64228-81-5
demonstration of compliance. See also 5.10. Control of impurities Action and use
in substances for phannaceutical use) A J D. Non-depolarizing neuromuscular blocker.
PhE" _
DEFINITION
.Mixture of the cis-cis, cis-trans and trans-trans isomers of
2,2 '- [pentane-I,5-diylbis[oxy(3-oxopropane-1,3-diyl)]]bis[1-
(3,4-dimelboxybenzyl)-6,7-dimethoxy-2-methyl-I,2,3,4-
tetrahydroisoquinolinium] dibenzenesulfonate.
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2022 Atracurium Besilate 1-229
Content potassium dihydrogen phosphate R previously adjusted to

96.0 per cent to 102.0 per cent (anhydrous substance). pH 3.1 with phosphoric acid R;
PRODUCTION - mobile phase B: mix 20 volumes of acetonitrile R,
It is considered that alkyl benzenesulfonate esters are 30 volwnes of methanol Rand 50 volumes of a 10.2 gIL
solution of potassium dihydrogen phosphate R previously
genotoxic and are potential impurities in atracurium hesitate.
adjusted to pH 3.1 with phosphoric acid R;
The manufacturing process should he developed taking into
together with considerations of the quality of starting (min) (per cent VIJI) (per cent VIJI)
materials) process capability and validation. The general
0-5 80 20
method 2.5.41. Methyl, ethyland isopropyl benzenesulfonate in
j.15 80 --> 40 20 --> 60
activesubstances is available to assistmanufacturers.
15·25 40 60
CHARACTERS 25 - 30 40 ..... 0 60 --> 100
Appearance 30 - 45 0 [00
White or yellowish-white, slightly hygroscopic powder.
Solubility Flow rate I mlzmlo.
Soluble in water) very soluble in acetonitrile, in ethanol Dueaion Spectrophotometer at 280 om.
(96 per cent) and in methylene chloride. Injection 20 J1L of test solution (a) and reference
IDENTIFICATION solutions (a), (h), (d) and (e).
A. lofrared absorption spectrophotometry (2.2.24). Identification of in/pun'ties Use the chromatogram obtained
Comparison atracurium besiJate CRS. with reference solution (d) and the chromatogram supplied
B. Examine the chromatograms obtained in the assay. with atra<urium for peak identification CRS to identify the
peaks due to impurities AI, A2, B, CI, C2, DI, D2) E, G
Results The 3 principal isomeric peaks in the chromatogram
and K; use the chromatogram obtained with reference
obtained with test solution (a) are similar in retention time to
solution (e) and the chromatogram supplied with atraounum
those in the chromatogram obtainedwith reference
for impurity F identification CRS to identify the peak due to
solution (a).
impurity F.
TESTS Relative retention With reference to the atracurium cis-cis
Soludon S isomer (retention time = about 30 min):
Dissolve 1.00 g in waterR and dilute to 100 mL with the impurity E = about 0.2; impurity F = about 0.25;
same solvent. ' =
impurity G about 0.3; impurity D I about 0.45; =
Appearance of solution impurity D2 = about 0,5; atracurium lmns-
Solution S is clear (2.2.1) and not more intenselycoloured trans isomer = about 0.8; atracurium cis-
than reference solution Y1 (2.2.2, Method 11). trans isomer = about 0.9; impurity AI = about 1.04;
Related substances =
impurity II about 1.07; impurity HI about 1.07 =
Liquid chromatography (2.2.29). (shoulder on the front of peak A2); impurity A2 (major
= =
isomer) about 1.08; impurity KI about 1.09 (shoulder
on the tail of peak A2); impurity 12 (major
examined in mobile phase A and dilute to 50.0 mL with isomer) = about 1.12; impurity H2 (majorisomer) ;;;; about
mobile phase A. 1.12; impurity K2 (majorisomer) = about 1,12;
Test soiution (b) Dissolve O.IOOg of the substance to be impurity B = about 1.15; impurity CI ;;;; about 1.2j
examined in mobile phase A and dilute to 10.0 mL with impurity C2 (majorisomer) = about 1.3.
mobile phase A. Systemsuitability:
Reference solution (a) Dissolve 50.0 mg of atraam'um - resolution: minimum J.5 between the peaks due to the
besilate CRS in mobile phase A and dilute to 50.0 mL with atracurium trans-trans isomer and the atracurium tis-trans
mobile phase A. isomer, and minimum 1.5 between the peaks due to the
Reference solution (b) Dilute 1.0 mL of test solution (a) to atracurium cis-trans isomer and the atracuriurn cis-cis
100.0 mL with mobile phase A. isomer in the chromatogram obtained with reference
Reference solution (c) Dissolve 20.0 mg of methyl solution (a);
beneenesulfonate R in acetonitrile R and dilute to 100.0 mL - peak-to-valley ratio: minimum 1.2, where Hp = height
with the same solvent. Dilute 50 J1L of the solution to above the baseline of the peak due to impurity Al and
100.0 mL with mobile phase A. H IJ = height above the baseline of the lowest point of the
Reference solution (d) Dissolve 2.0 mg of aO'a<urium for peak curve separating this peak from the peak due to the
identification CRS (containing impurities AI, A2, B, CI, C2, atracurium cis-cis isomer in the chromatogram obtained
D I, D2, E, G and K) in 2.0 mL of mobile phase A. with reference solution (d).
Reference solution (e) Dissolve 2.0 mg of atracurium for Limits:
impurity F identification CRS in 2.0 mL of mobile phase A. - correction factor. for the calculation of content, multiply the
peak area of impurity G by 0.5;
Column: - impmity E: not more than 1.5 times the sum of the areas
- size: 1= 0.25 m, 0 = 4,6 nun;
of the peaks due to the atracurium cis-cis, trans-trans and
- stationary phase: base-deactivated end-eapped octad«y/silyl cis-trans isomers in the chromatogram obtained with
silica gd for ehromarography R (5 pm). reference solution (b) (1.5 per cent);
Mobile phase: - impurities A, D: for each impurity, for the sum of the areas
- mobile phaseA: mix 5 volumes of methanol R, 20 volumes of the 2 isomer peaks, not more than 1.5 times the sum
of acetonitrile Rand 75 volumes of a 10.2 gIL solution of of the areas of the peaks due to the atracurium cis-cis,
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1-230 Atracurium Besilate 2022
trans-trans and cis-trans isomers in the chromatogram Sulfated ash (2.4. If)
obtained with reference solution (b) (1.5 per cent); M.aximum 0,1 per cent, determined on J.O g.
- impun"lY C: for the sum of the areas of the 2 isomer peaks,
ASSAY
not more than me sum of the areas of the peaks due to
the atracurjum cis-cis, trans-trans and cis-trans isomers in
(1.0 per cent); Injection Test solution (a) and reference solution (a).
- impuniies P, G: for each impurity, not more than the sum Calculate the percentage content of C6~82N2018S2 from
of the areas of the peaks due to the atracurium ds-cis, the sum of the areas of the peaks due to the 3 isomers.
lrans-trans and cis-trans isomers in the chromatogram STORAGE
obtained with reference solution (b) (1.0 per cent); In an airtight container, protected from light, at a
- impurities H;, I, K: for the sum of the areas of the isomer temperature of 2 DC to 8 DC.
peaks of these impurities, not more than the sum of the
areas of the peaks due to the atracurium cis-cis, trans-trans IMPURITIES
and cis-trans isomers in the chromatogram obtained with Specified ;mpun·tie.s A, G, D, E, F, G, H, I, J. K.
reference solution (b) (1.0 per cent); Other detectable impurities (the following substances would, if
- unspeaJied impuniies: for each impurity, not more than present at a sufficient level. be detected by one or otherof the tests
0.1 times the sum of the areas of the peaks due to the in the monograph. They are limited by the general acuplance
atracurium cis-eis, trans-trans and cis-trans isomers in the criterion for otherlunspuified impun'ties andlor 1!)' the general
chromatogram obtained with reference solution (b) monograph Substances for phannaceulical use (2034). It is
(0.10 per cent); therefore not necessary ro i'denufy these impuriu·es for
- rotal: not more than 3.5 times the sum of the areas of the demonstration of compliance. See also 5.10. Control of impurities
peaks due to the atracurium tis-cis, trans-trans and cis-Irons in substances for pharmaceutical use) B.
isomers in the chromatogram obtained with reference
- disregard limit:. 0.05 times the sum of the areas of the
peaks due to the atracurium tis-cis, trans-trans and cis-trans
isomers in the chromatogram obtained with reference
solution (b) (0.05 per cent).
impurity J
Uquid chromatography (2.,2.29) as described in the test for
Mobile phase:
Tim, MobUe phase A Moblle phase B

(min) (per cent VA? (per cent VIII)
0·5 80 20
5 - 15 80 -+ 75 20 -+ 25
15 - 25 75 25 A. 1-(3,4-dimethoxybenzyl)-2-[13-[I-(3,4-dimethoxybenzyl)-
25 - 30 75 -+ 55 25 -+ 45 6,7 -dimethoxy-3,4-dihydroisoquinolin-2( IH}-yl]-3,11-
30 - 38 55 -+ 0 45 -+ 100 dioxo-4, I o-dioxanidecyl]-6,7-dimethoxy-2-methyl-I,2,3,4-
38 - 45 0 100 tetrahydroisoquinolinium (Al : : ;: trans isomer, A2 ::::;: cis
isomer),
Injection 100 ~L of test solution (b) and reference
solution (c).
Retention time Impurity J : : ;: about 25 min; atracuriurn trans-
trans Isomer e about 38 min.
Limit:
- impurity J: not more than the area of the principal peak in
the chromatogram obtained with reference solution (c)
(10 ppm).
Isomer composition
related substances with the following modification. Use the
normalisation procedure.
Injection Test solution (a).
Limits:
B. pentane-I,S-diyl bis[3-[I-(3,4-dimethoxybenzyl)-6,7-
- atracunum cis-cis isomer: 55.0 per cent to 60.0 per cent,
dimethoxy-3,4-dihydroisoquinolin-2(1H}-yl]propanoateJ,
- atraaeium cis-trans isomer. 34.5 per cent to 38.5 per cent,
- atraam'um trans-trans isomer: 5.0 per cent to 6.5 per cent
Water (2.5.12)
Mexlmum 5.0 per cent, detennined on 1.000 g.
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2022 Atracurium Besilate 1-231
H'CO~
I 0
~
C. 1-(3,4-dimeth oxybenzyl)-2-(3, l l-dioxo-t, I O-dioxatridec- HCO 0
12-enyl)-6,7-dimethoxy-2-methyl-I,2,3,4- H;CO '" ••

rerrahydroisoquinolinium (Ct :::: trans isomer, C2 :::: cis
I .N-CH,
isomer), H,CO
H. 2,2'-[hexane-I,6-diylbis [oxy(3-oxopropane-I, 3-diyl)] ]bis

[1-(3,4-dimethoxybenxyl)-6,7-dimethoxy-2-methyl-
1,2,3,4-tetrahydroisoquinoliniwn] (HI:::: cis-trans isomer,
H2 :::: cis-cis isomer),
D. 1_(3,4_dimethoxybenxyl)_2_[3_[(5_hydroxypentyl)oxy)-3-
oxopropyl)-6,7-dimethoxy-2-methyl-1 ,2,3 ,4-
terrahydroisoquinclinium (D 1 :::: trans isomer, D2 = cis
isomer),
I. 2,2'-[(3-methylpentane-I,5-diyl)bis[oxy(3-oxopropane-
E. 2-(2-carboxyethyl)-I-(3,4-dimethoxybenxyl)-6,7- I ,3-diy1))) bis[1-(3 ,4-dimethoxybenxyl)-6,7 -dimethoxy-2-
dimethoxy-2-methyl-l,2,3,4-tettahydroisoquinolinium, =
methyl-I,2,3,4-tetrahydroisoquinolinium) (II cis-trans
isomer, 12 :::: cis-cis isomer),
J. methyl benzenesulfonate,
F. 1_(3,4-dimethoxybenxyl)_6,7-dimethoxy-2,2-dimethyl-
l,2,3,4-tetrahydroisoquinolinium,
G. 1-(3,4-dimethoxybenxyl)-6,7-dimethoxy-2-methyl-I,2,3,4-
tetrahydroisoquinoline,
K. 2,2'-[hexane-I,5 -diylbis[oxy(3-oxopropane-1,3-diyl)lJbis
[1_(3,4-dimethoxybenxyl)-6,7-dimethoxy-2-methyl-
1,2,3 ,4-tetrahydroisoquinolinium] .
_ _ _ _ _~_ _~ ~ PhEII
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1-232 Atropine 2022
Dissolve 1.25 g in ethanol (96 per eenv R and dilute to

Atropine 25.0 mL with the same solvent.
(ph. Eur. monograph 2056) Related substances
examined in mobile phase A and dilute to 100.0 rnL with
andenanliomer mobile phase A.
solution to 10.0 mL with mobile phase A.
289.4 51-55-8 Reference solution (b) Dissolve 5 mg of atropine
impurity B CRS in the test solution and dilute to 20 mL with
Action and use the test solution. Dilute 5 rnL of this solution to 25 rnL with
Anticholinergic. mobile phase A.
PhE<I ~ _ Reference solution (c) Dissolve the contents of a vialof
atropine for peak identification CRS (containing impurities A,
DEFINITION D, B, F, G and H) in I mL of mobile phase A.
( IR,3R,5S)-8-Methyl-8-azabicyclo[3.2 .Ijocr- 3-yl (2RS)-3-
hydroxy-2-phenylpropanoate.
Reference ,olUlion (d) Dissolve 5 mg of tropic acidR
(impurity C) in mobile phase A and dilute to 10 mL with
Content mobile phase A. Dilute I mL of the solution to 100 mL with
99.0 per cent to 101.0 per cent (dried substance). mobile phase A. Dilute I mL of this solution to 10 mL with
CHARACTERS mobile phase A.
Appearance Column:
White or almost white, crystalline powder or colourless - size: 1= 0.10
ID J 0 = 4.6 mm;
crystals. srationary phase: polar end-capped ocradecy/silyl siJi<a gelfor
-
Solubility chromaUJgraphy R (3 urn).
Vety slightly soluble in water. freely soluble in ethanol Mobile phase:
(96 per cent) and in methylene chloride, - mobile phase A: dissolve 3.5 g of sodium dode<y/ sulfare R in
606 mL of a 7.0 gIL solution of potassium dihydragen
IDENTIFICATION
phosphare R previously adjusted to pH 3.3 with a 5.8 gIL
First idtntijiuztion: A, B, E.
solution of phosphoric acid R, and mix with 320 mL of
Second identification: A, C, D, E. acetonitrile Rl;
A. Melting point (2.2.14): 115 °C to 119 °C. - mobile phase B: aceUJnirrile Rl;
Comparison atropine CRS. Tim, Moblle phase A MobIle phase B
(min) (per cent VIJI) (per cent YM
95 5
Test solution Dissolve 10 mg of the substance to be 95...., 70 5...., 30
examined in methanol R and dilute to 10 mL with me same
solvent.
Flow rare I mUmin.
Reference solution Dissolve 10 mg of atropine CRS in
lnjeaion 10 ~L.
Plare TLC silica gelplare R.
Mobile phase concentrated ammonia R, water R, acetone R with atropine for peak identification CRS and the
(3:7:90 VIVIV).
Application I0 ~L. the peaks due to impurities AJ D J EJ FJ G and H; use the
Development Over half of the plate. chromatogram, obtained with reference solution (b) to
Drying At 100-105 °C for 15 min. identify the peak due to impurity B; use the chromatogram
Detection After cooling) spray with dilute potassium obtained with reference solution (d) to identify the peak due
Wdobismuthare SolUlion R. to impurity C.
Results The principal spot in the chromatogram obtained Relative retention With reference to atropine (retention
with the test solution is similar in position, colour and size to time = about II min): impurity C = about 0.2;
the principal spot in the chromatogram obtained with the impurity E = about 0.67; impurity D = about 0.73;
reference solution. =
impurity F about 0.8; impurity B about 0.89; =
impurity H = about 0.93; impurity G = about 1.1;
D. Place about 3 mg in a porcelain crucible and add 0.2 mL
ofjuming nitric addR. Evaporate to dryness on a water-bath,
=
Dissolve the residue in 0.5 mL of a 30 gIL solution of System suitability Reference solution (b):
potassium hydroxide R in methanol R; a violet colour develops. - resolution: minimum 2.5 between the peaks due to
impurity B and atropine.
E. Optical rotation (see Tests).
Limits:
TESTS - correction faaon: for the calculation of content, multiply
Optical rotation (2.2.7) the peak areas of the following impurities by the
-0.700 to + 0.05 0 (measured in a 2 dID rube).
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2022 Atropine Sulfate 1-233
H
impurity C = 0.6; r"l ~ HJ--b
- impurities E, H: for each impurity, not more chan 3 times ~o,···\",H. . H andepimeralC'
obtained with reference solution (a) (0.3 per cent); \,~ H OH
- impurities A, B, C, D, F, G: for each impurity, not more
than twice the area of the principal peakin the E. (IS,3R,5S,6RSJ-6-hydroxy-8-methyl-8-azabicyclo[3.2.I]
chromatogram obtained with reference solution (a) oet-3-yl (2SJ-3-hydroxy-2-phenylpropanoate
(0.2 per cent); (7-hydroxyhyoscyamine),
- unspedfied impurities: for each impurity, not more than the
- totol: not more than 5 times the area of me principal peak
in the chromatogram obtained with referencesolution (a)
(0.5 per cent);
the chromatogram obtained with reference solution (a) F. (lR,2R,4S,5S, 7')-9-methyl-3-oxa-9-azatricyclo[3.3.I.O'·'j
(0.05 per cent). non-7-yl (2SJ-3-hydroxy-2-phenylpropanoate (hyoscine),
H
ASSAY
~>C~~~:~~;] aodenennomer
Dissolve 0.250 g in 40 mL of anhydrous aatic acid R, heating

if necessary, and allow to cool Titrate with 0.1 M perchloric
acid, determining the end-point potentiometrically (2.2.21J). G. (lR,3r,5SJ-8-methyl-8-azabicyclo[3.2.I]oct-3-yl (2RSJ-2-
1 mL of 0.1 M perchloric acid is equivalent to 28.94 mg hydroxy-3-phenylpropanoate (littorine),
of C11H23N03' H. unknown structure.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'"
STORAGE
IMPURITIES
Specified impun'lies A, B, C, D, E, P, G, H.
Atropine Sulfate ***
*** ***
""00H~"---------]
A
.6'
0
O'
• N-CH3
•........
Atropine Sulphate
***
~
cy~~N_~_C~~]~
CH2 H
I "" 0 --.......
A. (IR,3r, 5SJ-8-methyl-8-azabicyclo[3. 2.1]oct-3-yl
2-phenylpropenoate (apoatropine),
H . H,SO,. H,O
IH~"']
OH 2 and enanliomer
()
~ ( : -o-'''-_/~_ and enenucmer 695 5908-99-6
OH H Action and use

Anticholinergic.
B. (IR,3r,5SJ-8-azabicyclo[3.2.I]oct-3-yl (2RSJ-3-hydroxy-2- Preparations
phenylpropanoate (ncratropine), Atropine Eye Drops
AtropineEye Ointment
~
I ..,? ...~
co H
2 and enanliomer
Atropine Injection
Atropine OealSolution
OH Atropine Tablets
PhE", _
C. (2RSJ-3-hydroxy-2-phenylpropanoic acid (tropic acid),
DEFINITION
?H
u; ~
H
Bis[(1R,3r,5SJ-8-methyl-8-azabicyclo[3.2.1]oct-3-yl (2RSJ-3-
I 0 --- . H
..,?.. :~... N-CH; and epimerat C'
hydroxy-2-phenylpropanoate] sulfate monohydrate.
Content
H 99.0 per cent to 101.0 per cent (anhydrous substance).
"OHH
CHARACTERS
D. (IR,3S,5R,6RSJ-6-hydroxy-8-methyl-8-azabicyclo[3.2.I] Appearance
oet-3-yl (2SJ-3-hydroxy-2-phenylpropanoate White or almost white, crystalline powder or colourless
(6-hydroxyhyoscyamine), crystals.
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1-234 Atropine Sulfate 2022
Solubility Time Mobile phase A MobUe phase B

(min) (per cent VIII) (per cent VIII)
Very soluble in water, freely soluble in ethanol (96 per cent).
95 5
IDENTIFICATION
95 --> 70 5 --> 30
First idenhJicati<m: A, B, E.
Second identjficatian: C, D, E, F. Flow rate 1 mllmin.
A. Optical rotation (see Tests).
Deteaion Spectrophotometer at 210 nm.
Injection 10 flL.
Comparison ..ropine sulfate CRS. Identification of impurities Use the chromatogram supplied
C. Dissolve about 50 mg in 5 mL of waterR and add 5 mL with arropine for peak idemijication CRS and the
of picric acidsolution R. The precipitate, washed withwater R chromatogram obtainedwith reference solution (c) to identify
and dried at 100-105"C for 2 h, melts (2.2.14) at 174 "C to the peaks due to impurities A, D, E, F, G and H; use the
179 "C. chromatogram obtainedwith reference solution (b) to
D. To about I mg add 0.2 mL ofjuming nitric acid Rand identify the peak due to impurity B: use the chromatogram
evaporate to dryness in a water-bath. Dissolve the residue in obtainedwith reference solution (d) to identify the peak due
2 mL of acewne R and add 0.1 mL of a 30 gIL solution of to impurity C.
potassium hydroxide R in methanol R. A violet colour develops. Relative retention With reference to atropine (retention
E. It gives the reactions of sulfates (2.3. I). time = about 11 min): impurity C =about 0.2j
F. It gives the reaction of alkaloids (2.3.1). impurity E = about 0.67; impurity D = about 0.73;
impurity F = about 0.8; impurity B = about 0.89;
TESTS
pH (2.2.3)
impurity H = =
about 0.93; impurity G about 1.1;
4.5 to 6.2.
System suilability Reference solution (b):
Dissolve 0.6 g in earbon dWxide-free waterR and dilute to - resolution: minimum 2.5 between the peaks due to
30 mL with the same solvent. impurity B and atropine.
Optical rotation (2.2. 7) Limits:
-0.50" to + 0.05" (measured in a 2 dm tube). - correction factors: for the calculation of content, multiply
Dissolve 2.50 g in water R and dilute to 25.0 mL with the the peak areas of the following impurities by the
same solvent. corresponding correction factor: impurity A = 0.6;
Related substances impurity C = 0.6;
Liquid chromatography (2.2.29). - impun·ties E) H: for each impurity) not more than 3 times
obtainedwith reference solution (a) (0.3 per cent),
- impurities A, B) C) D) F) G: for each impurity) not more
mobile phase A.
than twice the areaof the principal peak in the
Reference solution (a) Dilute 1.0 mL of the test solution to chromatogram obtained with reference solution (a)
100.0 mL with mobile phase A. Dilute 1.0 mL of this (0.2 per cent);
solution to 10.0 mL with mobile phase A. - unspecified impuruies: for each impurity, not more than the
Reference solu'ion (b) Dissolve 5 mg of atropine area of the principal peak in the chromatogram obtained
impurilJ! B CRS in the test solution and dilute to 20 mL with with reference solution (a) (0.10 per cent);
the test solution. Dilute 5 mL of this solution to 25 mL with - total: not more than 5 times the area of the principal peak
mobile phase A. in the chromatogram obtained with reference solution (a)
Reference solution (c) Dissolve the contentsof a vial of (0.5 per cent);
atropine forpeak idellIificau'on CRS (containing impurities A, - disregard limit: 0.5 times the area of the principal peak In
D, E, F, G and H) in I mL of mobile phase A. the chromatogram obtained with reference solution (a)
Reference solution (d) Dissolve 5 mg of tropic acid R (0.05 per cent).
(impurity C) in mobile phase A and dilute to 10 mL with Water (2.5.12)
mobile phase A. Dilute I mL of the solution to 100 mL with 2.0 per cent to 4.0 per cent, determined on 0.500 g.
mobile phase A. Dilute I mL of this solution to 10 mL with Sulfated ash (2.4.14)
mobile phase A. Maximum 0.1 per cent, determined on 1.0 g.
Column:
ASSAY
- size: I =0.10 m, 0 = 4.6 mmj
Dissolve 0.500 g in 30 mL of anhydrous acetic acid R,
- slationary phase: polarend-capped ocwde<ylsi!YI silka gelfor
warming if necessary. Cool the solution. Titratewith 0.1 M
chromawgraphy R (3 urn).
perchloric acid, determining the end-pointpotentiometrically
Mobile phase: (2.2.21J).
- mobile phaseA: dissolve 3.5 g of sodium dode<yl sulfate R in
I mL of 0.1 M perchlotic acid is equivalent to 67.68 mg
606 mL of a 7.0 gIL solution of potassium dihydrogen
of C3..u,aNzOIOS.
phosphate R previously adjusted to pH 3.3 with a 5.8 gIL
solution of phosphork acidR, and mix with 320 mL of STORAGE
euewnitrile R1j Protected from light.
- mobile phase B: acetonitrile Rlj IMPURITIES
Specified impurities A) B) C) D) E)F) G) H.
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2022 Attapulgite 1-235
Attapulgite
Action and use
Excipient.
DEFINITION
A. (I R,3r,5S)-8-methyl-8-azabicyclo[3.2.1 Joct-3-y)
Attapulgite is a purified native hydrated magnesium
2-phenylpropenoate (apoatropine),
aluminium silicate essentially consisting of the clay mineral
H
palygorskite.
'-"'" (:
~-'-']
() 1'~'·\..-r.
OH H
and enanUomer
CHARACTERISTICS
A light, cream or buff, very fine powder, free or almostfree
from gritty particles.
IDENTIFICATION
A. Ignite0.5 g with 2 g of anhydrous sodium carbonate for
B. (IR,3r,5S)-8-azabicyclo[3.2.1]oct-3-yl (2RS)-3-hydroxy-2- 20 minutes, cool and extract with 25 mL of boiling water.
phenylpropanoate (noratroplne), Cool, filter, wash me residue wiili water and add the
washings to the filtrate. Reserve the residue for test B.
~
Cautiously acidify the combined filtrate and washings with
t COH hydrochloric acid, evaporate to dryness, moisten the residue
..# -OH'2 andenanliomer
with 0.2 mL of hydrochloric add, add 10 mL of water and stir.
OH
A white, gelatinous precipitate is produced.
B. Wash the residue reserved in test A with water and
C. (2RS)-3-hydroxy-2-phenylpropanoic acid (tropic acid), dissolve in 10 mL of 2M hydrochloric acid. To 2 mL of the
solution add a 10% wlv solution of ammonium thiocyanate.
An intensered colour is produced.
C. To 2 mL of the solution obtained in test B add 1 mL of
strong sodium hydroxide solution and filter. To the filtrate add
3 mL of ammonium chlonae solution. A gelatinous white
precipitate is produced.
D. To 2 mL of the solution obtained in test B add
D. (IR,3S,5R,6RS)-6-hydroxy-8-methyl-8-azabicyclo[3.2.1] ammonium chloride and an excess of 13.5M ammonia and
oct-3-yl (2S)-3-hydroxy-2-phenylpropanoate filter. To the filtrate add 0.15 mL of magneson reagent and an
(6-hydroxyhyoscyamine), excess of 5M sodium hydroxide. A blue precipitate is produced.
TESTS
~ ~HJ-__ ~H
Acidity or alka1lnlty
pH of a 5% w/v suspension in carbon dioxide-free water, after
~o····\ ... N-CH~ ...-H andepimeralC· shaking for 5 minutes, 7.0 to 9.5, Appendix V L.
. H
Adsorptive capacity
'elH H OH
Moisture adsorption, 5 to 14% when determined by the
following method. Dry in air and powdera sufficient quantity
E. (IS,3R,5S,6RS)-6-hydroxy-8-methyl-8-azabicyclo[3.2.1] of the substance being examined and pass through a sieve
oct-3-y! (2S)-3-hydroxy-2-phenylpropanoate with a nominal mesh aperture of 150 um. Spread 0.5 g as a
(7-hydroxyhyoscyamine), thin layer on a previously weighed piece of aluminium foil
(60 mm x 50 mm) of nominal gauge 17.5 urn and transfer
~ ~ J __ ~H rH
to a desiccator containing a dish of sodium chloride crystals
partially immersed in saturated brine at 25°. After 4 hours,
~:_'\_._ N-CH3, 0 removefrom the desiccator and weigh immediately. Dry in
H . an oven at 1100 for 4 hours, allow to cool in a desiccator and
\ H H
OH weigh. The moisture adsorption is the gain in weightof the
substance being examined expressed as a percentage of its
F. (IR,2R,4S,5S, 7,)-9-methyl-3-oxa-9-azatricyclo[3.3.I.O"'] oven-dried weight.
non-7-yl (2S)-3-hydroxy-2-phenylpropanoate (hyoscine), Arsenic
To 0.13 g add 5 mL of water, 2 mL of mlfuric acid and
10 mL of sulfur dioxide soluu"on and evaporate on a water bath
until the sulfur dioxide solutionis removed and the volume
reduced to about 2 mL. Transfer the solution to the
generator flask with the aid of 5 mL of water. The resulting
solution complieswith the limit tes: for arsenic, Appendix vn
(8 ppm).
G. (lR,3r,5S)-8-methyl-8-azabicyclo[3.2.1] oct-3-yl (2RS)-2-
hydroxy-3-phenylpropanoate (Iirtorine), Acid-solebte matter
H. unknown structure. Boil 2 g with 100 mL of 0.2M hydrochloric acid under a reflux
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>EII
condenserfor 5 minutes, cool and filter. Evaporate 50 mL of
the filtrate to dryness. The residue, after ignition at about
600° for 30 minutes, weighs not more than 0.25 g.
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1-236 Attapulgite 2022
Water-soluble matter the filtrate to dryness. The residue, after ignition at about
Bail 10 g with 100 mL of water under a reflux condenserfor 600° for 30 minutes, weighs not more than 0.25 g.
5 minutes, cool and filter. Evaporate 50 mL of the filtrate to Water-soluble matter
dryness. The residue, after ignition at 600 0 for 30 minutes, Boil 109 with 100 mL of water under a refluxcondenser for
weighs not more than 50 mg. 5 minutes, cool and filter. Evaporate 50 mL of the filtrate to
Loss on drying dryness. The residue, afterignition at 600° for 30 minutes,
When dried to constant weight at 105°, loses not more than weighs not more man 50 mg.
17.0% of its weight. Use I g. Adsorptive capacity
Loss on ignition In a stoppered bonie shake 1.0 g, in very fine powder, with
When ignited at 600°, loses 15.0 to 27.0% of Its weight. 50 mL of a 0.12% wlv solution of methylene blue for
Use I g. 5 minutes, allow to settle and centrifuge. The colour of the
clear supernatant solution is not more intense man that of a
0.0012% wlv solution of methylene blue.
Loss on drying
Activated Attapulgite When dried to constant weight at 105°, loses not more than
Action and use 4.0% of its weight. Use I g.
Antidiatthoeal. Loss on ignition
When ignited at 600°, loses not more than 9.0% of its
DEFINITION weight. Use I g.
Activated Attapulgite is a purified native hydrated magnesium
aluminium silicate essentially consisting of the claymineral
palygorskite that has been carefully heated to increase its
adsorptive capacity. Azathioprine
CHARACTERISTICS
A light, cream or buff, very fine powder, free or almost free (Ph. Eur. monograph 0369)
from gritty particles.
N-{No,
IDENTIFICATION
A. Ignite 0.5 g with 2 g of anhydrous sadium carbona" for <~
N s
20 minutes, cool and extract with 25 mL of boiling water.
H3t A-.-~
Cool, filter, wash the residue with waler and add the
washings to the filtrate. Reserve the residue for test B.
Cautiously acidifythe combined filtrate and washings with
t.J-- >
N N
hydrochb>ric add, evaporate to dryness, moisten the residue
with 0.2 mL of hydrochloric acid, add 10 mL of waW and stir. G,H,N,O,s 277.3 446-86-6
A white, gelatinous precipitate is produced.
Action and use
B. Wash the residue reserved in test A with water and
dissolve in 10 mL of 2M hydrochloric acid. To 2 mL of the
Immunosuppressant.
solution add a 10% wlv solution of ammonium thiocyanate. Preparations
An intense red colour is produced. Azathioprine Oral Suspension
C. To 2 mL of the solution obtained in test B add I mL of Azathioprine Tablets
strong sodium hydroxide solution and filter. To the filtrate add PIlE" _
3 mL of ammonium chloride solution. A gelatinous white
precipitate is produced. DEFINITION
D. To 2 mL of the solution obtained in test B add 6- [( l-Methyl-d-nitro-I H-imidazol-5-yl)sulfanyl)-7H-purine.
ammonium chloride and an excess of 135M ammonia and Content
filter. To the filtrate add 0.15 mL of magneson reagent and an 98.5 per cent to 101.0 per cent (dried substance).
excess of 5M sodium hydroxide. A blue precipitate is produced.
CHARACTERS
TESTS Appearance
Acidity or a1kallnity Pale-yellow powder.
pH of a 5% wlv suspension in carbon dioxide-free wartr, after Solubility
shaking for 5 minutes, 7.0 to 9.5, Appendix V L Practically insoluble in water and in ethanol (96 per cent).
Arsenic It is soluble in dilute solutions of alkali hydroxides and
To 0.13 g add 5 mL of wa,.r, 2 mL of sulfuric acid and sparingly soluble in dilute mineral acids.
10 mL of sulfur dioxide solution and evaporate on a water bath
IDENTIFICATION
until the sulfur dioxide solution is removed and the volume
reduced to about 2 nIL Transfer the solution to the
generator flask with the aid of 5 mL of water. The resulting Comparison azathioprine CRS.
solution complies with the limit test for arsenic, Appendix vn TESTS
(8 ppm). Related substances
Acid-soluble matter Liquid chromatography (2.2.29).
Boil 2 g with 100 mL of 0.2M hydrochwric acid under a reflux Solution A 2.76 gIL solution of sadium dihydrogen phospha"
condenser for 5 minutes, cool and filter. Evaporate 50 mL of monohydrate R adjusted to pH 2.5 with phosplwric acidR.
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2022 Azathioprine 1-237
Test solution Dissolve 10 mg of the substance to be Loss on drying (2.2.32)

examined in 35 mL of a 0.8 gIL solution of sodium Maximum 1.0 per cent, determined on 0.500 g by drying in
hydroxide R and dilute to 100.0 mL with solution A. an oven at 105°C.
Reference solution (a) Dissolve 5 mg of azathiopn"ne Sulfated ash (2.4.14)
impurityA CRS and 5 mg of mercaptopurine monohydrate R Maximum 0.1 per cent, determined on 1.0 g.
(impurity B) in 8.15 mL of a 0.8 gIL solution of sodium
ASSAY
hydroxide. R and dilute to 25.0 mL with solution A.
Dissolve 0.250 g in 25 mL of dimethylformamide R. Titrate
To 1.0 mL of this solution, add 35 mL of a 0.8 gIL solution
with 0.1 M tetrabutylammonium hydroxide, determining the
of sodium hydroxide R and dilute to 100.0 mL with
solution A.
Reference solution (b) Dissolve 2.5 mg of ozachioprine 1 mL of 0.1 M tetralnttylammonium hydroxide is equivalent to
21.13 mg of C.H,N,02S.
impun"ty G CRS and 2.5 mg of the substance to be examined
in 8.8 mL of a 0.8 gIL solution of sodium hydroxide Rand STORAGE
dilute to 25.0 mL with solution A. To 1.0 mL of this Protected from light.
solution, add 17.5 mL of a 0.8 gIL solution of sodium
IMPURITIES
hydroxide R and dilute to 50.0 mL with solution A.
Specified impurUies A, B.
Ocher detectable impurities (thefollowing substances would, if
100.0 mL with solution A. Dilute 1.0 mL of this solutionto
present at a SUfficienllevelJ be detected by oneor other of the tests
in the monograph. Theyare limited by the general acceptance
Column: criterion for OIherlumpec-ified impurities and/or by the general
- size: 1 = 0.15 m, (2) = 4.6 rnm; monograph Substances for pharmaceutical use (2034). II is
- stationary phase: phenylsilyl silica gelfor chromaUJgraphy R therefore not necessary to identify these impurities for
(5 urn); demonstration 0/compliance. See also5.10. Concrol of impurities
- temperature: 30 "C. in substances for pharmaceutical use) C) D, E, F G. J
Mobile phase:
- mobile phase A: methanol R, solution A (5:95 VIV); N NO,
- mobile phase B: solution A, methanol R (40:60 VIV); <J( ~ NH:z
Time Mobile phase A MobUe phase B H,C
(min) (per cent JlIY) (per cent VIP)
0-5 100 0 A. l-methyl-4-rutro-IH-imidazol-5-amine,
5·15 100 -i 0 0-->100
l5 - 20 0 100 SH
N~~
t.,Jl
N
>
N
Injection 20 ~L. B. 7H-purine-6-thiol (mercaptopurine),

Identification of impuniies Use the chromatogram obtained
with reference solution (a) to identify the peaks due to N NO,
impurities A and B. Use the chromatogram obtainedwith <-,(
reference solution (b) to identify the peak due to impurity G. ,
NACI
Relative retention With reference to azathioprine (retention H,C
tirne > about 15 min): impurity A :::: about 0.3;
impurity B = about0.4; impurity G :::: about 0.97. C. 5 -chloro-1-methyl-4-nitro-lH-imidazole,
System SUiUlhiJity:
impurities A and B in the chromatogram obtainedwith
reference solution (a); minimum 2.0 between the peaks
due to impurity G and azathioprine in the chromatogram
obtained with reference solution (b).
D. l-methyl-4-rutro-IH-imidazole-5-thiol,
Limits:
- impun'{ies A) B: for each impurity) not more than
(X
NO,
chromatogram obtainedwith reference solution (c)
(0.15 per cent); , N OH
H,C
- unspecified impurides: for each impurity, not more than the
with reference solution (c) (0.10 per cent); E. l-rnethyl-a-nitro-Uf-Imidazol-Sol,
- total: not more than 5 times the area of the principal peak o
:J: >
H
(0.5 per cent); HN N
- disregard limir. 0.5 times the area of the principal peakin t., N I N
(0.05 per cent). F. 1,1-dihydso-6H-purin-6-one (hypoxanthine),
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1-238 Azelastine Hydrochloride 2022

solutionto 10.0 mL with the solventmixture.
Reference solution (b) Dissolve I mg of benzohydrazide R
(impurity A), 1 mg of azelastine impurity B CRS, I mg of
2-[2-(4-ch/orophenyl)awy/]benzoic acid R (impurity C), I mg
of azelastine impurily D CRS and 1 mg of azelastine
G. 6-[(I-methyl-4-nitro-IH-imidazol-5-yl)sulfanyl]-7H-purin- impurily E CRS in the test solution and dilute to 20.0 mL
2-amine (thiamiprine). with the test solution.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'1>&
Column:
- size: I;;;: 0.25 m, 0 ;;;: 4.6 mm;
- stationary phase: cyan0Sl7y1 silica gelfor chromatography R
(5 urn):
Azelastine Hydrochloride - temperature: 30 "C.
Mobile phase Dissolve 2.16 g of sodium octanesu/fonate Rand
(Ph. Eur. monograph 1633) 0.68 g of potassium dihydrogen phosphate R in 740 mL of water
for chromatography R; adjust to pH 3.0-3.1 with dilute
o;D,c~
""N
phosphoric acid R, add 260 mL of autom·trile R1 and mix.
. Hel
Injection I0 ~L.
andenanUomer . Run time 1.5 times the retention time of azelastine.
CI Identification of impuriuts Use the chromatogram obtained
C",H"CI,N,O 418.4 79307-93-0 impurities A, B, C, D and E. The elution order may vary,
especially for impurity E, but the peakarea of each
Action and use impurity in reference solution (b) is different, so a clear
HistamineHI receptor antagonist; antihistamine. identification of the impurities is possible.
I'I>E~ _ Relative retention With reference to azelastine (retention
DEFINlTION
4-[(4-Chlorophenyl)methyl)-2-[(4RS)-I-methylhexahydro-
= =
impurity B about 0.3; impurity C about 0.4;
impurity D = about 0.6; impurity E = about 0.8.
IH-.,epin-4-yljphthal.,in-1 (21f)-one hydrochloride.
System suitabaily:
Content - resolution: minimum 2.0 between the peaks due to
99.0 per cent to 101.0 per cent (dried substance). impurity E and azelastine and minimum 1.5 between the
CHARACTERS peaks due to impurities C and D in the chromatogram
Appearance obtained with reference solution (b);
White or almost white, crystalline powder. - signal-eo-noise ratio: minimum 90 for the principal peak in
the chromatogram obtained with reference solution (a).
Solublllty
Sparingly soluble in water, soluble in anhydrous ethanol and Calculation ofpercentage ccnUnts:
in methylene chloride. - correction faaon: multiply the peak areas of the following
IDENTIFICATION impurity A = 2.6; impurity B =4.5; impurity C = 2.0;
A. Infrared absorption spectrophotometry (2.2.24). impurity E = 2.8;
Comparison aselastine hydrodlloride CRS. - for each impurity, use the concentration of azelastine
B. Solution S (see Tests) gives reaction (a) of chlorides hydrochloride in reference solution (a).
(2.3.1). Limits:
TESTS
- imptmues AJ B, C, E: for each impurity, maximum
0.15 per cent;
Solution S
- unspecified impun'ties: for each impurity, maximwn
0.10 per cent;
- total: maximum 0.2 per cen1;
Acidity or alkalinity - reporting threshold: 0.05 per cent.
solution RI. Not more than 0.1 mL of 0.01 M hydrochloric
acid or 0.01 M sadium hydroxide is required to change the
an oven at 105°C.
colourof the indicator.
liquid chromatography (2.2.29). In order to awid overheating in the reaction medium, mix
thoroughly throughout and stop the titration immediately afterthe
SoIwnt mixture acetonitrile Rl, waterfor chromatography R
end-point has been reached.
(45:55 VIV).
Dissolve 0.300 g in 5 mL of anhydrous formic acidR.
Add 30 mL of acetic anhydride R. Titrate quickly with 0.1 M
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2022 Azithromycin 1-239
perchlotic acid.. determining the end-point potentiometrically ***

Azithromycin
(2.2.20). *** ***
1.0 mL of 0.1 M perchloric acid is equivalent to 41.84 mg of (Ph. Eur. monograph 1649) ***
C22H25ChN30.
IMPURITIES
Specified impurities A, B, C, E.
Otherdetectable impurities (the following substances WQUld, if
present at a sufficient level.. be detected by om or other of the tests
criterion for othetiunspedfied impurities and/or by me general
monograph Sub,wn<es for pharmaceutical use (2034). It is
therefore not necessary l() identify these impuniies for
demonstration of eompdance. See also5.10. Control of impun"ties
in substances for pharmaceutical use) D.
Cj8H72N20.z,XH20 749 (anhydrous substance)

with x e lor2
A. benzohydrazide (benzoyldiazane), Azilhromycin monohydrate 121470-24-4

Azithromycin dihydrate 117772-70-0
and enanliomer Action and use
Macrolide antibacterial.
Preparations
B. N' -[(4RS)-I-methylhexahydro-IH-azepin-4- Azithromycin Capsules
yl]benzohydrazide,
'2Y
Azithromycin Eye Drops
Azithromycin for Infusion
CO
Azithromycin Oral Suspension
"" 0' H
Azithromycin Tablets
PhE,; _
~I
DEFINITION
a "" (2R,3S,4R,5R,8R, lOR,IIR, 12S, 13S, 14R)-13-[(2,6-Dideoxy-
3-G-methyl-3-0-methyl-«-L-rib<>-hexopyranosyl)oxy]-2-ethyl-
C. 2-[2-(4-chlorophenyl)acetyl]benzoic acid, 3,4, 10-trihydroxy-3,5,6,8,I 0, 12, 14-heptamethyl-II-[[3,4,6-
trideoxy-3- (dimethylamino) -f!-n-xy/<>-hexopyranosyl] oxyj-t-
0
oxa-S-azacyclopentadecan-If-one. The degree of hydration is
~ NH lor 2.
I
",N Semi-synthetic productderived from a fermentation product.
"" Content
~
a
"" CHARACTERS
Appearance
D.4-[(4-chlorophenyl)methyl]phthalazin-I(2H)-one, White or almost white powder.
Solubillty
o Practically insoluble in water, freely soluble in anhydrous
ethanol and in methylene chloride.
IDENTIFICATION
and (l}-isomer Infrared absorption spectrophotometry (2.2.24).
Comparison azithromycin CRS.
prepare further spectra using 90 gIL solutions in methylene
chloride R.
E. (3E)-3-[(4-chlorophenyl)methylidene]-2-benzofuran-1
TESTS
(3H)-one.
Solution S
_ _ _ _ _ _ _ _ _ _ _ _ _ _~ PhE,;
Dissolve 0.500 g in ~nhydrou, ethanol R and dilute to
Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
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1-240 Azithromycin 2022
pH (2.2.3) = =
impurity 0 about 1.23; impurity G about 1.26;
9.0 [0 11.0. =
Dissolve 0.100 g in 25.0 mL of methanol R and dilute to System suitability Reference solution (b):
50.0 mL with carbon dioxide-free waterR. - peak-to-valley ratio: minimum 1.4, where Hp = height
Specific optical rotation (2.2.7) above the baselineof the peak due to impurity Jand
-49 to -45 (anhydrous substance), determined on solution S. HI} = height above the baseline of the lowest point of the
Related substances
impurity F.
Limits:
Solvent mixture Prepare a 1.73 gIL solution of ammonium - correaion jCUf(JTS: for the calculation of content, multiply
dihydrogen phosphate R adjusted to pH 10.0 with ammonia R. the peak areas of the following impurities by the
Transfer 350 mL of this solution to a suitable container. corresponding correction factor: impurity F = 0.3;
Add 300 mL of acetonitrile Rand 350 mL of methanol R.
impurity G = 0.2; impurity H =0.1; impurity L = 2.3;
Mix well. = =
impurity M 0.6; impurity N 0.7;
Test solution Dissolve 0.200 g of the substance to be - impun"ty B: not more than Mice the area of the principal
examined in the solvent mixture and dilute to 25.0 mL with peak in the chromatogram obtained with reference
the solventmixture. solution (a) (2.0 per cent);
Reference solution (a) Dilute 1.0 mL of the test solution to - impurities A, C, E, F, H, I, L, M, N, 0, P: for each
100.0 mL with the solvent mixture. impurity, not more than 0.5 times the area of the
Reference solution (b) Dissolve the contents of a vial of principal peak in the chromatogram obtainedwith
azimromy<in for system suitability CRS (containing impurities reference solution (a) (0.5 per cent);
F, Hand D in 1.0 mL of the solvent mixture and sonicate - sum oj ;mpun'ties D, J and Q: not more than 0.5 times the
for 5 min. area of me principal peakin me chromatogram obtained
Reference solution (c) Dissolve 8.0 mg of azimromy<in for with reference solution (a) (0.5 per cent);
puzk identification CRS (containing impurities A, B, C, E, F, - impun"ty G: not more than 0.2 times the area of the
G, I, J, L, M, N, 0 and P) in 1.0 mL of the solvent mixture.
Column: - any other impuniy: for each impurity, not more than
- size: / = 0.25 m, 0 = 4.6 nun; 0.2 times the area of the principal peak in the
- stationary phase: end-capped octadecylsilyl amorphous chromatogram obtained with reference solution (a)
organosilica polymer for chromatography R (5 1'IIl); (0.2 per cent);
- temperature: 60°C. " - total: not more than 3 times the area of the principal peak
Mobile phase: in the chromatogram obtained with reference solution (a)
- mobile phase A: 1.80 gIL solution of anhydrous disodium (3.0 per cent);
hydrogen phosphate R adjusted to pH 8.9 with dilute - disregard limit: 0.1 times the area of the principal peak in
phosphoric acidR or with dilute sodium hydroxide solution R; the chromatogram obtained with reference solution (a)
- mobile phase B: memanolR2, acetonitrile Rt (25:75 VII'); (0.1 per cent); disregard the peaks eluting before
impurity L and alier impurity B.
Time MobUe phase A Mobile phase B
(min) (per cent VIJ.? (per cent V/J?
Water (2.5.12)
0-25 50 -+ 45 50 -+ 55
25 - 30 45 -+ 40 55 -+ 60 Sulfated ash (2.4.14)
30 - 80 40 -+ 25 60 -+ 75 Maximum 0.2 per cent, determined on 1.0 g.
80 - 81 25 -+ 50 75 -+ 50 ASSAY
81·93 50 50 Liquid chromatography (2.2.29).
Solution A J\lix 60 volumesof acetonitrile R and 40 volumes
Flaw rate 1.0 mUmin. of a 6.7 gIL solution of dipotassium hydrogen phosphate R
Detection Spectrophotometer at 210 run" adjusted to pH 8.0 with phosphoric acidR.
Injection 50 11L. Test solutUm Dissolve 53.0 mg of the substance to be
ldenujication 0/impun"ties Use the chromatogram supplied examined in 2 mL of acetonitrile R and dilute to 100.0 mL
with azimromycin for peak identification CRS and the with solution A.
chromatogram obtained with reference solution (c) to identify Reference solution (a) Dissolve 53.0 mg of azithromy<in CRS
the peaks due to impurities A, B, C, E, F, G, I, J, 1., M, N, in 2 mL of acetonitrile R and dilute to 100.0 mL with
o and P; use the chromatogram supplied with azithromycin solution A.
for system suitability CRS and the chromatogram obtained Reference solution (b) Dissolve 5 mg of the substance to be
with reference solution (b) to identify the peakdue to examined and 5 mg of azithromycin impurity A CRS in
impurity H. 0.5 mL of acetonitrile R and dilute to 10 mL with solution A.
Relative retention With reference to azithromycin (retention Column:
time = 45-50 min): impurity L = about 0.29; - size: 1 = 0"25 ID, 0 = 4.6 nun;
= =
impurity M about 0.37; impurity E about 0.43; - stationary phase: octadecylsilyl vinyl polymer for
= =
impurity F about 0.51; impurity D about 0.54; chromatography R (5 ~m);
impurity J = about 0.54; impurity Q = about 0.54; - temperature: 40°C.
impurity I = about 0.61; impurity C = about 0.73;
Mobile phase Mix 60 volumes of acetonitrile Rl and
= =
impurity N about 0.76; impurity H about 0.79;
40 volumes of a 6.7 gIL solution of dipotassium hydrogen
= =
impurity A about 0.83; impurity P about 0.92;
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2022 Azithromycin 1-241
phosphate R adjusted to pH 11.0 with a 560 gIL solution of

potassium hydroxide R.
Injection I0 ~L.
Run time 1.5 times Ute retention time of azithromycin.
Retention time Azithromycin = about 10 min.
System suitabiiliy Reference solution (b):
impurity A and azithromycin.
Calculate the percentage content of C3sHnN2012 taking into D. 14-demethyl-14-(hydroxymethyl)azithromycin
account the assigned content of Qzilhromycin CRS. (aziiliromycin F),
STORAGE
CH,
l..:1-
IMPURITIES
Specified impurities A, B, C, D, E, F, GJ H, I, J, L, M, N,
0, P, Q.
in the monograph. Theyare limited by the general a«eptance
ca
7
~H: HO~:~ /:H,oHCH,
OH
c~
OCH3
H
H
\
CH3
0
therefore not newsary to identify these ;mpuril;es for CH,
demonstration of compliance. See also5.10. Control of impun"ties
in substances for phannaceuticaJ use) K. E. 3'-(N,N-didemethyl)azithromycin (aminoazithromycin),
A. 6-demethylazithromycin,
F. 3'-N-demethyl-3'-N-fonnylazithromycin,
B. 3-deoxyazithromycin (azithromycin B),
G. 3'-N-demethyl-3'-N-[(4-methylphenyl)
sulfonyl]azithromycin,
C. 3"-Q-demethylazithromycin (azithromycin C),
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2022
1-242 Azithromycin
H,C
H···
H,C
CH3 HO'"
OHC~H °1---H-t;'
'L--..( H~01
OH? HCH3 HI '.
CH,
H. 3'_N_[[4_(acetylamino)phenyljsulfonyl]-3' -N- M.3'_(N,N_didemethyl)-3'-N-fonnylazithromycin,

demethylazithromycin, CH,
H,C
H···· CH, H~:~~ H:13~H,
;--O~ Ho~i H"
. ,~'O~CH'
H,C
CH) HO
)-oo---+--
H3C(~H 'j Ho~i }--{
o
CH)
CH3
0
'L--..( CH3 H
QH OCH3 H
OH OCH3
CH,
CH,
N. 3'-de(dimethylamino)-3'-oxoazithromycin,
I. 3/_N_demethylazithromycin,
CH,
~3
I
H,C N
H'"
• H
H,C OH
CH3 HO···· I CH3
h o o'-----J-.. H
H,cqH~01H'
OH?
CH,
J. 13-o-decladinosylazithromycin,
O. 2_desethyl_2_propylazithromycin,
H,C P. unknown structure,
H"
H,C
HO" H,C
H·
H,C
o 0:;- 0 0
CH3 HO'"
D -'<4 H~\r
8
o v I NH OH
H,C)lN ",'
H OH? Ii
CH,
K. CI \ l "-epoxyazithromycin (azithromycin E),
Q.3'_N_[[4_(acetylamino)phenYl)sulfonylj-3'-(N,N-
didemethyl)azithromycin.
_ _ _~ P>E"
o
H'~
L. azithromycin 3'-N-oxide,
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2022 Bacampicillin Hydrochloride 1-243
contents of the tube by swirling; the solutionis practically

Bacampicillin Hydrochloride colourless. Place the test-tube on a water-bam for 1 min;
(Ph. Eur. monograph 0808) a darkyellow colour develops.
D. Dissolve about 25 mg in 2 mL of walerR. Add 2 mL of
dilute sodium hydroxide solution R and shake. Walt a few
minutes and add 3 mL of dilute nit"'c acid Rand 0.5 mL of
silver nitrate solution R1. A white precipitate is formed,
Add 0.5 mL of concentrated ammonia R. The precipitate
dissolves.
TESTS
502.0 37661-08-8 Appearance of solution
Dissolve 0.200 g in 20 mL of water Rj the solution is not
Action and use more opalescent than reference suspension IT (2.2.1).
Penicillin antibacterial. Dissolve 0.500 g in 10 mL of water R; the absorbance
(2.2.25) of the solution at 430 nm is not greater than 0.10.
PhE.. ~~ _
pH (2.2.3)
DEFINITION 3.0 to 4.5.
(IRSJ-I-[(Ethoxycarbonyl}oxy)ethyl (2S,5R,6R}-6-[[(2R}-2- Dissolve 1.0 g in carbon dioxide-free water R and dilute to
amino-2-phenylacetyl] amino)-3,3-dimethyl-7 -oxo-a-thia-I- 50 mL with the same solvent.
azabicyclo[3.2.0)heptane-2-carboxylale hydrochloride.
Semi-synthetic product derived from a fermentation product. + 175 to + 195 (anhydrous substance).
Content Dissolve 0.250 gin waW R and dilute to 25.0 mL with the
95.0 per cent to 102.0 per cent (anhydrous substance). same solvent.
CHARACTERS Related substances
Appearance liquid chromatography (2.2.29). Prepare the test solution and
White or almost white powder or granules, hygroscopic. reference solutions (a), (b) and (d) immediately before use.
Solubility Phosphate buffer A Dissolve 1.4 g of sodium dihydrogen
Soluble in water, freely soluble in ethanol (96 per cent), phosphate monohydrate R in water R and dilute to about
soluble in methylene chloride. 800 mL with the same solvent. Adjust to pH 3.0 with dilute
phosphori<: acidR and dilute to 1000.0 mL with w""" R.
IDENTIFICATION
Firstidentification: A, D. Phosphate bufferB Dissolve 2.75 g of sodium dihydrogen
phospha", mononydrau Rand 2.3 g of disodium hydrogen
phospha", dihydra'" R in waterR and dilute to ahout 1800 mL
A. Infrared absorption spectrophotometry (2.2.24). with the same solvent. Adjust to pH 6.8, if necessary, using
Comparison bacampiciOin hydrochloride CRS. dilu'"phosphoric acidR or dilu",sodium hydroxide solution R
B. Thin-layer chromatography (2.2.27). and dilute to 2000.0 mL with wa"'r R.
Testsolution Dissolve 10 mg of the substance to be Test solution Dissolve 30.0 mg of the substance to be
examined in 2 mL of methanol R. examined in phosphate buffer A and dilute to 100.0 mL with
Reference sdution (a) Dissolve 10 mg of bacampiciOin phosphate buffer A.
hydrochloride CRS in 2 mL of methanol R. Reference solution (a) Dissolve 30.0 mg of bacampicillin
Reference solution (b) Dissolve 10 mg of bacompicillin hydrochloride CRS in phosphate buffer A and dilute to
hydrochloride CRS, 10 mg of la/ampicillin hydrochloride CRS 100.0 mL with phosphate buffer A.
and 10 mg of pivampiciOin CRS in 2 mL of methanol R. Reference solution (b) Dilute 1.0 mL of reference solution (a)
Pia", TLC sikmised silica gelpia'" R. to 100.0 mL with phosphate buffer A.
Mobile phase Mix 10 volumes of a 272 gIL solution of Reference solution (c) Dissolve 30 mg of the substance to be
sodium acetate R adjusted to pH 5.0 with glacial acetic acid R, examined in phosphate buffer B and dilute to 100 mL with
40 volumes of water Rand 50 volwnes of ethanol phosphate buffer B. Heat at 80°C for about 30 min.
(96 per cem) R. Reference solution (dJ Dissolve 20 mg of ampicillin
Appliwtion I ~L. trihydrate CRS (impurity I) in phosphate buffer A and dilute
to 250 mL with phosphate buffer A. Dilute 5 mL of this
solution to 100 mL with phosphate buffer A..
Drying In a current of warm air.
Column:
Detection Spray with ninhydn·n solution Rl and heat at 60°C - size: 1= 0.05 m, Q) = 3.9 mmj
for 10 min. - stationary phase: oetadecylsilyl silka gelfor chromaUJgraphy R
System suitability Reference solution (b): (5 urn).
- the chromatogram shows 3 clearly separated spots. ll1.obile phase 1\1ix 30 volumes of acetonitrile Rl and
Results The principal spot in the chromatogram obtained 70 volwnes of a 0.06 per cent mlm solutionof
with the test solution is similar in position, colour and size to tetrahexylammonium hydrogen sulfate R in phosphate buffer B.
the principal spot in the chromatogram obtainedwith Flow rate 1.0 mllmin.
reference solution (a). Detection Spectrophotometer at 220 om.
C. Place about 2 mg in a test-tube about 150 mm long and lnj«tion 20 J.lL of the test solutionand reference
15 mm in diameter. Moisten with 0.05 mL of water Rand solutions (b), (c) and (d).
add 2 mL of sulfuric acid-formaldehyde reagent R. Mix the
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1-244 BacampicilIin Hydrochloride 2022
Run time 3.5 times the retention time of bacampicillin.

Syuem miUlbility:
- the peakdue to impurity I is separated from the peaks
due to the solvent in the chromatogram obtained with
reference solution (d);
B. (2R)-2-amino-2-phenylacetic acid (o-phenylglycine),
- relative retention with reference to bacampicillin:
degradation producteluting justafter
=
bacampicillin 1.12 to 1.38 in the chromatogram
obtained with reference solution (e); if necessary, adjust
the concentration of tetrahexylammonium hydrogen
sulfate in the mobile phase.
Limits:
·
en ,
-.. I
H NH2
0
HN
~ ~s.3
'"!
I. )<CH
COzH
X-- CH3
- Q,-ry impun"ty: for each impurity, not more than 1.5 times
C. (2RS,4S)-2-[([(2R)-2-amino-2-phenylacetyl]
the area of the principal peak in the chromatogram amino]methyl]-S,S-<iimethylthiarolidine-4-carboxylic acid
obtained with reference solution (b) (1.5 per cent); (penilloic acids of ampicillin),
in the chromatogram obtained withreference solution (b)
(3 per cent);
the chromatogram obtained withreference solution (b)
(0.1 per cent).
Butyl acetate and ethyl acetate (2.4.24, System A)
Maximum 2.0 per cent of butyl acetate, maximum D. (4S)-2-[([(2R)-2-amino-2-phenylacetyl]
4.0 per cent of ethyl acetate and maximwn 5.0 per cent for aminolcarboxymethyl]-S,5-dimethylthiarolidine-4-
the sum of the contents. carboxylic acid (penicilloic acids of ampicillin),
Sample solution Dissolve 50.0 mg of the substance to be
solvent.
Use the method of standard additions.
SUllie head-space conditions .thatmay be used:
- equilibration temperature: 60°Cj
- equilibration time: 20 min.
N,N-Dimethylaniline (2.4.26, Metlwd A)
Maximwn 20 ppm.
E. (4S)-2-(3,6-dioxo-S-phenylpiperazin-2-yl)-5,S-
Water (2.5.12) dimethylthiawlidine-4-carboxylic acid (diketopiperazines
Maximum 0.8 per cent, determined on 0.300 g. of ampicillin),
HS CH3
ASSAY
HeX
.3 ~ X~~H and enanUomer
H Nt<,
F. (2RS)-2-amino-3-methyl-3-sulfanylbutanoic acid (OL-
Injection Test solution and reference solution (a). penicillamine),
- repeatabiHty: maximum relative standard deviation of
Calculate me percentage content of C2IH2sCIN.301S from
the declared content of ba<ampiciUin hydrochloride CRS.
STORAGE G. methyl (2R)-2-amino-2-phenylacetate (methyl
In an airtight container. p-phenylglycinate),
IMPURITIES
H
O)---~J(~:3
~N····fi-S'><CH3
H H
A. (2S,SR,6R) -ti-amino-3,3-<iimethyl-7-oxo-d-thia-I»
H. (IRS)-I-[(ethoxycarbonyl)oxy]ethyl (2S,SR,6R)-6-[[(2R)-
2-(acetylamino)-2-phenylacetyl] amino]-3,3-dimethyl-7-
(6-aminopenicillanic acid), oxo-4-thia-l-azabicyclo[3.2.0]heptane-2-carboxylate (N-
acetylbacampicillin),
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2022 Bacitracin 1-245
Solubility
Freely soluble in water and in ethanol (96 per cent),
IDENTIFICATION
First identification: B, C.
Second idendfication: Ai C.
I. (2S,5R,6R)-6-[[(2R)-2-amino-2-phenylacetyl]amino]-3,3- A. Thin-layer chromatography (2.2.27).
dimethyl-7-oxo-4-thia-I-azabicyclo[3.2.0]heptane-2- Testsolution Dissolve 10 mg of the substance to be
carboxylic acid (ampicillin). examined in a 3.4 gIL solution of hydro<hlorit: acid Rand
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE«
dilute to 1.0 mL with the same solution.
Reference solution Dissolve J0 mg of bacitracin zinc CRS in a
3.4 gIL solution of hydro<hloric acidR and dilute to 1.0 mL
Bacitracin Plate TLC silica gelplat< R.
Mobile phase glacial acetic add R, water R, buumol R
(14:29:57 VIV/V).
Applit.,ion 10 ~L.
H~X
Hl
s ~N
R
Development Over half of the plate.
Drying At 100-105 "C.
Detection Spraywith ninhydn'n solution R1 and heat at
~·.ll-leu- D-Glu- Y-t-Lys'" o-Om -- X-- o-Ph9J 110 °C for 5 min.
o t L-Asn-- o-Asp--l-His Results The spots in Ute chromatogram obtained with the
test solution are similar in position, size and colour to the
Name Mol. Formula X Y R
spots in the chromatogram obtained with the reference
BacitracinA C66HIOCINI70ISS l-ila c-ue
CH, solution.
Bacilracin BI C&sHl01NI701~ L-Ile L-Ile
H B. Composition (see Tests).
Bacitracin 82 C&sHl01N110t6S l-Val l-lle CH,
C. Ignite 0.2 g. There is no significant yellow-coloured
Bacitracin B3 C~IOlNI7016S L-lle l·Val CH,
residue at high temperature. Allow to cool. Dissolve the
residue in 0.1 mL of daut< hydrochloric acid R. Add 5 mL of
1405-87-4 water R and 0.2 mL of strong rodium hydroxide sO/alUm R.
No white precipitate is formed.
Action and use
TESTS
Polypeptide antibacterial.
Solution S
PIIE« _ Dissolve 0.25 g in carbon dioxide-free water R and dilute to
DEFINITION
Mixtureof antimicrobial polypeptides produced by certain Appearance of solution
strains of Bacillus lichenifonnis or Badllus subti/is, the main Solution S is clear (2.2.1).
components being: pH (2.2.3)
- 4,10-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2- 6.0 to 7.0 for solution S.
methylbutyl]-4,5-dihydro-I,3-thiazol-4-yl] carbonyl]-L- Composition
leucyl-o-«-glutamyl-L-isoleucyl-L-Iysyl-o-omithyl-L- Liquid chromatography (2.2.29): use the normalisation
isoleucyl-o-pheuylalanyl-L-histidyl-D-«-aspartyl-L- procedure. Prepare thesolutions immediately before use.
asparagine] (bacitracin A);
Solution A 40 WL solutionof sodium edeuzte R adjusted to
- 4,10-anhydro[N-[[(4R)-2-[(IS)-I-amino-2-methylpropyl]-
pH 7.0 with dilute sodium hydroxide solution R.
4,5 -dihydro-I,3-thiazol-4-yl] carbonyl]-L-Ieucyl-o-«-
glutamyl-L-isoleucyl-L-Iysyl-I>-omithyl-L-isoleucyl-I>- Solution B In a volumetric flask) dissolve 54.4 g of potassium
phenylalanyl-L-histidyl-D-«-aspartyl-L-asparagine] dihydrogen phosphate R in wate,for throma",graphy Rand
(bacitracin HI); dilute to 2000 mL with the same solvent. Adjust to pH 6.0
- 4,10-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2- with a 34.8 gIL solution of dipotassium hydrogen phosphate R
methylbutyl]-4,5-dihydro-1,3-thiazol-4-yl] carbonyl]- L- and filter through a membrane filter (nominal pore size
leucyl-o-«-glutamyl-L-isoleucyl-L-Iysyl-o-omithyl-L-valyl- 0.45 prn),
I>-phenylalanyl-L-histidyl-o-«-aspartyl-L-asparagine] Test solution Dissolve 0.100 g ofthe substance to be
(bacitracin B2); examined in the mobile phase and dilute to 50.0 mL with
- 4,10-anhydro[N-[[(4R)-2-[(lS,2S)-I-amino-2- the mobile phase.
methylbutyl]-4,5-dihydro-1,3-thiazol-4-yl] carbonyl]-L- Reference solution (a) Dissolve 20.0 mg of bacitracin for
leucyl-o-«-glutamyl-L-valyl-L-Iysyl-o-omithyl-L-isoleucyl- system suitability CRS in solution A and dilute to 10.0 mL
I>-phenylalanyl-L-histidyl-o-«-aspartyl-L-asparagine] with the same solution.
(bacitracin B3). Reference solution (b) Dilute 5.0 mL of reference solution (a)
Content to 100.0 mL with solution A. Dilute 1.0 mL of this solution
Minimum 60 ill/mg (dried substance). to 10.0 mL with solution A.
CHARACTERS Reference solution (c) In order to prepare impurities E, F, G
Appearance and H in mu, heat about 4 mL of reference solution (a) in a
White or almost white, hygroscopic powder. water-bath for 30 min. Cool to room temperature.
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1-246 Bacitracin 2022
0.20 10
0.15
~0.10 4
5
0.05
0.00
0 5 10 15 20 25 30 35 40 45 so 55 60 min
0.010 8
0.008
0.006
7
~0.004
0.002 12 16
0.000 13 14 15
-0.002
0 5 10 15 20 25 30 35 40 45 so 55 60 min
I. impurity A 5. bacitracin 82 9. impurityL 13. impurity F

2. impurity B 6. bacitracin 83 10. bacitracin A 14. impurity G
3. impurityC 7. impurityM u. impurity 0 15. impurity H
4. bacitracin BI 8. impuriry N 12. impurities P and Q 16. impurity E
Figure 0465.-1. - Chromatogram for the testfor cemposiu'on of bacitracin: lestsolution
Column: System suitability:

- size: 1:= 0.15 m, 0 = 4.6 mm; - peak-UHJalley ratio: minimum 1.2, where Hp = height
- seaeWnary phase: end-eapped, charged sur/au, ethylene-bridged above the baseline of the peak due to bacitracin H2 and
octadecylsi/yl silica gelfor chromawgraphy (hybrid materiaV R H v = heightabove the baseline of the lowestpoint of the
(3.5 urn), curve separating this peakfrom the peakdue to bacitracin
- temperature: 28 ± 2 "C. Bl in the chromatogram obtained with reference
Mobile phase acetonitrile R, solution B, waterfor solution (a);
chromatagmphy R, methanol RJ (43:100:300:557 VIVIVII'). - peak-/(J-fJalley meW: minimum J.J, where Hp = height
Flaw mte 1.0 mllmin. above the baseline of the peak due to impurity M and
H; = heightabove the baseline of the lowest point of the
Detection Spectrophotometer at 254 run. curve separating this peak from the peak due to bacitracin
Injection 100 ~L of the test solution and reference B3 in the chromatogram obtained with reference
solutions (a) and (b). solution (a);
Run time 3 times the retention time of bacitracin A - signal-to-noise ratio: minimwn 50 for the peak due to
Identification of peaks Use the chromatogram obtained with bacitracin A in the chromatogram obtained with reference
reference solution (a) to identifythe peaks due to impurity M solution (b).
and bacitracins A, B1, B2 and B3 (see Figure 0465.-1). Limits:
Relative retention With reference to bacitracin A (retention - bacitracin A: minimum 45.0 per cent;
= =
time about 20 min): impurity A about 0.44; - sumof bacitracins A.,Bl, B2 and B3: minimum
= =
impurity B about 0.52; impurity C about 0.55; 77.0 per cent;
bacitracin HI = about 0.65; bacitracin B2 = about 0.67; - reporting threshold: 0.25 per cent.
= =
bacitracin B3 about 0.81; impurity Iv! about 0.87; Related substances
impurity N = about 0.90; impurity L = about 0.93; Liquid chromatography (2.2.29) as descnbed in the test for
impurity 0 =about 1.2; impurities P and Q =about 1.3; composition with the following modifications. Use the
= =
impurity F about 1.6; impurity G about 1.8; normalisation procedure.
= =
impurity H about 2.1; impurity E about 2.8. Injution Test solution and reference solutions (a), (b) and
If necessary, adjust the composition of the mobile phase by (c).
changing the amount of organic modifier whilstkeeping the Identification of jmpun"ties Use the chromatogram obtained
ratio constant between methanol and acetonitrile. with reference solution (a) to identify the peaks due to
impurities A, B, C, L, J\[, N, 0, P and Q (see
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2022 Bacitracin 1-247
1 2
3
-0.01O-h~--r;rr-~.-r~~,..,--r;rr-,......~",..,~--r;rr-rr-~.-r~,..,~~rr-~.-r~,..,~--r;rr-~
o 10 15 20 25 30 35 40 45 50 55 min
1. impurity F 2. impurity G 3. impurity H 4. impurity E
Figure 0465.-2. - Chromatogram for the test for relaid substances of bacitracin: reference solution (c)
Figure 0465.-1); use the chromatogram obtained with

reference solution (c) to identify the peaks due to
impurities E, F, G and H (see Figure 0465.-2).
:rvt'
,l CH 3
S ~N
Limits:
- sum of impurities L and N: maximum 8.0 per cent; y.."~t-Leu-o-Glu -- t-lIe-t·lys -o-Ofn--t-Val-o-PheJ
- impm;ty E: maximum 4.0 per cent; H 0 It-Asn-o-AsP-t-HiS
- impun"ty A: maximum 3.5 per cent;
- impurities B~ M: for each impurity, maximum 3.0 per cent; A. 4,1 O-anbydro[N-[[(4R)-2-[ (I SJ-I-amino-2-methylpropyl]-
- impun"ty C: maximwn 2.5 per cent; 4,5-dihydro-l,3-thiazol-4-yl]carbonyl]-L-Ieucyl-b-«-
- sum 0/impun"ties 0, P and Q: maximum 2.5 per cent; glutamyl-L-isoleucyl-L-lysyl-D-omithyl-L-valyl-D-
- sum of impun"ties F and G: maximum 2.0 per cent; phenyJ,lanyl-L-histidyl-D-o:-asparryl-L-asparagine]
- impurity H: maximum 1.0 per cent; (bacitracin DI, bacitracin C2),
- airy OIlier impun"ty: for each impurity, maximum
2.0 per cent;
H2~vt"
- reporting threshold: 0.25 pet cent. Hl CH3
Loss on drying (2.2.32) s ~N
Maximum 5.0 per cent, determined on 1.000 g by drying in ~"'~ r-t.eu-e- o-eiu- t-Va/-t-Lys-- o-Om-e-c-ne -- o-PheJ
vacuo at 60 °C at a pressure not exceeding 0.1 kPa for 3 h. o tt.Asn-o-Asp-t-Hls
Maximum 1.0 per cent, determined on 1.0 g. B. 4,10-anhydro[N-[[(4R)-2-[(ISJ-I-amino-2-methylpropyl]-
ASSAY 4,5 -dihydro-I,3-thiazol-4-yl] carbonylj-t-leucyl-n-a-
Carry out the microbiological assay of antibiotics (2.7.2). glutamyl-L-valyl-L-lysyl-D-omithyl-L-isoleucyl-D-
Use bacitracin zinc CRS as the reference substance. phenylalanyl-L-histidyl-D-o:-asparryl-L-asparagine]
(bacitracin D2, bacitracin C3),
STORAGE
In an airtight container at 2 °C to 8°C" If the substance is
sterile, the container is also sterile and tamper-evident.
IMPURITIES
Specified impurities AJ BJ CJ EJ FJ OJ HJ LJ MJ N, 0, P, Q.
presenr at a sufficient level, bedetected by oneor otherof the tests
criterion for other/unspecified impun"lies. It is therefore not C. 4, Io-anbydro[N-[[(4R)-2-[(I S,2SJ-I-amino-2-
necessary to identify these impun"ties for demonstratiun of methylbutyl] -4,5-<1ihydro-l ,3-thiazol-4-yl]carbonyl]-L-
compliance. See also 5.10. Control of impurities in substances for Jeucyl-D-o:-glutamyl-L-valyl-L-lysyl-D-omithyl-L-valyl-D-
pharma<eUtical use) D, I, J, K. phenylaJanyl-L-histidyl-D-«-asparryl-L-asparagine]
(bacitracin D3, bacitracin CIa),
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1-248 Bacitracin 2022
. D. 4,lO-anhydro[N-[[(4R)-2-[(IS)-I-amino-2-methylpropyl]-
4,5-dihydro-1,3-thiazol-4-yl]carbonyl]-L-leucyl-n-«- I. 4, I O-anhydro [N- [[ 2-(2-methyl-I-oxopropyl)-1,3-thiazol-4-
glutamyl-L-valyl-L-lysyl-o-omithyl-L-valyl-o-phenylalanyl- yl)carbonyl]-L-Ieucyl-o-«-glutamyl-L-isoleucyl-L-lysyl-o-
L-histidyl-o-«-aspartyl-L-asparagine) (bacitracin E), omithyl-L-valyl-o-phenylalanyl-L-histidyl-o-«-aspartyl-L-
asparagine) (bacitracin 11),
°r
CH,
CH
'
S N
>-"lOU- o-Glu- ,-Vol·,·lys -ec-om -'·110- o-PhO]

o tl-Asn-o-Asp-l-HiS
E. 4, IO-anhydro [N-[[2-[(2S)-2-methyl-I-oxobutyl]-1,3-
thiazol-4-yl]carbonyl]-L-Ieucyl-o-«-glutamyl-L-isoleucyl-L- J. 4,1 O-anhydro [N- [[2-(2-methyl-l-<>xopropyl)-l, 3-thiszol-4-
lysyl-o-omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-o-«- yl)carbonyl]-L-leucyl-o-«-glutamyl-L-valyl-L-lysyl-o-
aspartyl-t-asparagine] (bacitracin F), omithyl-L-isoleucyl-o-phenylaIanyl-L-histidyl-o-«-aspartyl-
L-asparagine] (bacitracin 12),
F. 4, IO-anhydro[N-[[2-(2-methyl-l-<>xopropyl)-1,3-thiazol-4-
K. 4, I O-anhydro[N-[[2-[(2S)-2-methyl-l-oxobutyl)-1 ,3-
yl]carbonyl)-L-Ieucyl-D-«-glutamyl-L-isoleucyl-L-lysyl-o-
thiazol-4-yl]carbonyl)-L-leucyl-o-«-glutamyl-L-valyl-L·lysyl-
omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-o-«-aspartyl-
o-omithyl-L-valyl-o-phenylalanyl-L-histidyl-D-«-aspartyl-L-
L-asparagine] (bacitracin HI),
asparagine] (bacitracin 13),
H CH
H~
Hi - CH 3
S "N
y... L-L..eU-o-GllI-Lolle-l-Lys-o-orn--L-llo,-o-PhO]
H [ ll.Asn-o-Asp_loHls
L. 4,IO-anhydro[N-[[(4R)-2-[(IR,2S)-I-amino-2-
G. 4,1O-anhydro [N-[[2-[(2S)-2-methyl-I-oxobutyl]-1,3- methylbutyl]-4,5-<1ihydro-1 ,3-thiazol-4-yl]carbonyl)-L-
thiazol-4-yl)carbonyl)-L-leucyl-o-«-glutamyl-L-isoleucyl-L- leucyl-O-O:-glutarnyl-L-isoleucyl-L-lysyl-o-omithyl-L-
Iysyl-o-omithyl-L-valyl-o-phenylalanyl-L-histidyl-D-«- isoleucyl-o-phenylalanyl-L-histidyl-o-«-aspartyl-L-
aspartyl-t-asparagine] (bacitracin H2), asparagine) (bacitracin X),
)~CH,
OJ,,.
S "N
>-'-l.U-o-GIU.,.V".,.lYS-o-O<n-,.II.-o-Phe
o t-Asn,,-o-AsP-l-HiS J
H. 4, IO-anhydro [N- [[2- [(2S) -g-methyl-l-oxoburylj-I,3- M.4,IO-anhydro[N-[[2-[(IS,2S)-I-amino-2-methylbutyl]-1,3-
thiazol-4-yl]carbonyl)-L-leucyl-o-«-glutamyl-L-valyl-L-lysyl- thiazol-4-yl]carbonyl)-L-leucyl-o-«-glutamyl-L-isoleuCyI-L-
o-omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-o-«- Iysyl-o-omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-D-«-
aspartyl-t-asparagine] (bacitracin H3), aspartyl-t-asparagine] (bacitracin Y),
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2022 Bacitracin Zinc 1-249
Bacitracin Zinc
a-
H~~X R
Hl
N.4,IO-anhydro[N-[[(4S)-2-[(IS,2S)-I-amino-2- s 'oN
methylbutyl]-4,5-<1ihydro-l,3-thiazol-4-yl]carbonyl]-L-
leucyl-o-<l-g1utamyl-L-isoleucyl-L-lysyl-o-ornithyl-L-
Y.1 r-teu -l>Glu- Y - c·Lys- 0-0,.- X- O-PheJ
isoleucyl-o-phenylalanyl-L-histidyl-o-<l-aspartyl-L- H 0 . tt.Asn ... o-Asp .... t.HiS

asparagine] (bacitracin Z),
N.me Mot. Formula X Y R
H,N H. CIi, Bacitracin A CeaHl(13NI10'8S L·11e t-lle CH,
H~CH' Bacllracin B1
Bacitracin 82
~HIOINI101l;S
CssHIOINI10,6S
L·lle L·lle H
L-Val L·lle CIi,
S N Bacitracin B-3 ~I01NI10u,S L-Ue lNal CH,
y. L-leU-o-Gfu .....M~---L-Lys--o-om-l.lle-o-PheJ
H r tt-Asn -o-Asp -L-His 1405-89-6
Action and use

O. Mil = 5-methylene-L-isoleucine; 4,IO-anhydro[N-[[(4R)-2-
Polypeptide antibacterial.
[(I S, 2S)-I-amino-2-methylbutyl]-4,5-dihydro-1,3-thiazol-
4-yl]carbonyl]-L-leucyl-D-<l-g1utamyl-5-methylene-L- Preparation
isoleucyl-L-lysyl-D-omithyl-L-isoleucyl-D-phenylalanyl-L- Polymyxin and Bacitracin Ointment
histidyl-n-c-aspartyl-t-asparagine] (bacitracin JI), Phf<l ~ _
DEFlNfTION
Zinc complex of bacitracin, which consists of a mixture of
antimicrobial polypeptides produced by certain strains of
Bacillus lkhenifonnis or Bacillus subtilis, the main components
being:
- 4,lo-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2-
methylb utyI]-4,5-<1ihydro-1,3-thiazol-4-yl]carbonyl]-L-
leucyl-D-<l-g1utamyl-L-isoleucyl-L-lysyl-D-ornithyl-L-
P. Mil = 5-methylene-L-isoleucine; 4,IO-anhydro[N-[(4R)-2- isoleucyl-D-phenylalanyl-L-histidyl-D-<l-aspartyl-L-
[(IS, 2S)-I-amino-2-methylbutyl]-4,5-dihydro-I,3-thiazol- asparagine] (bacitracin A);
4-yl]carbonylj-L-leucyl-o-<l-g1utamyl-L-isoleucyl-L-lysyl-D- - 4,lo-anhydro[N-[[(4R)-2-[(IS)-I-amino-2-methylpropyl]-
omithyl-5-methylene-L-isoleucyl-D-phenylalanyl-L-histidyl- 4,5-dihydro-1,3-thiazol-4-yl]carbonyl] -L-Ieucyl-n-«-
n-c-aspartyl-t-asperagine] (bacitracin J2), glutamyl-L-isoleucyl-L-lysyl-D-ornithyl-L-isoleucyl-o-
phenylalanyl-L-histidyl-D-<l-aspartyl-L-asparagine]
~~
(bacitracin Bl);
- 4,IO-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2-
~ 'CH 2
methylbutyl]-4,5-dihydro-1 ,3-thiazol-4-yl]carbonyl]-L-
S 'oN
leucyl-D-<l-g1utamyl-L-isoleucyl-L-lysyl-D-ornithyl-L-valyl-
y..
H r t-Leu-e- o-Glu--t·lle'" t-Lys- o-Orn -- L-lIe--O-PheJ
tL-Asn-o-Asp-t-HiS
o-phenylalanyl-L-histidyl-D-<l-aspartyl-L-asparaginej
(bacitracin B2);
- 4,IO-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2-
methylbutyl] -4, 5-dihydro-1,3-thiazol-4-yl] carbonylj-r-
Q.4,IO-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2-methylpem- leucyl-D-<l-g1utamyl-L-valyl-L-lysyl-D-ornithyl-L-isoleucyl-
4-en-l-yl]-4,5-dihydro-1,3-thiazol-4-yl]carbonyl] -t-leucyl- D-phenylalanyl-L-histidyl-D-<l-aspartyl-L-asparagine]
D-«-g1utamyl-L-isoleucyl-L-lysyl-D-ornithyl-L-isoleucyl-o- (bacitracin B3).
phenylalanyl-L-histidyl-D-<l-aspartyl-L-asparagine]
(bacitracin J3). Content
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf<l
Minimum 60 IU/mg (dried subsrance).
CHARACTERS
Appearance
White or light yellowish-grey, hygroscopic powder.
Solubility
Slightly soluble in water and in ethanol (96 per cent).
IDENTIFICATION
A. Thin-layer chromatography (2.2.27).
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1-250 Bacitracin Zinc 2022
0.20 10
0.15
~0.10 4
5
0.05
0.00
0 5 10 15 20 25 30 35 40 45 50 55 60 min
0.010 8
0.008
0.006
7
~0.004
0.002 12 18
0.000 1314 15
-0.002
0 5 10 15 20 25 30 35 40 45 50 55 60 min
1. impurityA 5. bacitracin 82 9. impurity L 13. impurity F

2. impurity B 6. beciuacin 83 10. bacitracin A 14. impurity G
3. impurity C 7. impurity M 11. hnpuriry 0 15. impurity H
4. hackracin 81 8. impurity N 12. impurities P and Q 16. impurity E
Figure 0466.-1. - ChromalCgram for <he USt for composition of btUitracin zinc: testsolution
Test solution Dissolve 10 mg of the substance to be Composition

examined in 0.5 mL of dilute hydrochloric add R and dilute '0 Liquid chromatography (2.2.29): use the normalisation
1.0 mL with water R. procedure. Prepa~ the solutWns immeJiauly before use.
Reference sdution Dissolve 10 mg of bacitracin zinc CRS in Solution A 40 gIL solution of sodium edetate R adjusted to
0.5 mL of dilute hydrochloric add R and dilute to 1.0 mL with pH 7.0 with dilutesodium hydroxid, solution R.
water R. Solution B In a volumetric flask, dissolve 54.4 g of potassium
Plate TLC silica gelplate R. dihydrogen phosphate R in waterfor chromawgraphy R and
Mobil, phase glacial acetic acid R, water R, butano/ R dilute to 2000 mL with the same solvent. Adjust to pH 6.0
(14:29:57 VIVIJI). with a 34.8 gIL solution of dipotassium hydrogen phosphate R
Application 10 ~L and filter through a membrane filter (nominal pore size
0.45 pm).
Development Over half of the plate.
Test solunOn Dissolve0.100 g of the substance [0 be
Drying At 100-105 "C. examined in solution A and dilute to 50.0 rnL with the same
Detection Spray with ninhydrin solution RI and heat a' solution.
110 "C for 5 min. Rejeren" solution (a) Dissolve 20.0 mg of badtracin for
Results The spots in the chromatogram obtained with the system suitability CRS in solution A and dilute to 10.0 mL
test solution are similar in position, size andcolour to the with the same solution.
spots in the chromatogram obtained with the reference Reference soluwn (b) Dilute 5.0 mL of reference solution (a)
solution. to 100.0 mL with solution A. Dilute 1.0 mL of this solution
B. Composition (see Tests). to 10.0 mL with solution A.
C. Ignite about 0.15 g, allow (Q cool anddissolve the residue Reference solution (c) In order to prepare impurities EJ F, G
in 1 mL of dilute hydrochloric acid R. Add 4 mL of waterR. and H in situ, heat about 4 mL of reference solution (a) in a
The solutiongives the reaction of zinc (2.3.1). water-bath for 30 min. Cool to room temperature.
TESTS Column:
pH (2.2.3) - size: 1= 0.15 rn, 0 = 4.6 mm;
6.0 to 7.5. - srationary phase: end-<apped, chorged sutface ,thylene-bridged
Shake 1.0 g for about 1 min with 10 mL of carbon dioxide-free oetadecy/silyl silica gelfor chromawgraphy (hybrid materiaD R
water R and filter. (3.5 pm);
- temperature: 28 ± 2 'C.
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0.080
0.070
0.06
0.04
~
0.030
0.020 4
0.010
1 2
3
0.0
I. impurity F 2. impurity G 3. impurity H 4. impurity E
Figure 0466.-2. - Chromatogram for the test/or related substances of bacitracin zinc: reference solution (cJ
Mobile phase acetonitrile R, solution B, waterjQ1' Limits:

chromarography R, methanol Rl (43:100:300:557 VIVIVIJI). - bacitracin A: minimum 45.0 per cent;
Flow rate 1.0 mllmin. - sum of baciuacins A, HI, B2 and B3: minimum
Detection Spectrophotometer at 254 nm. 77.0 per cent;
Injection 100 p.L of the test solution and reference
solutions (a) and (b). Related substances
Run lime 3 times the retention time of bacitracin A. Liquid chromatography (2.2.29) as described in the test for
Identification of peaks Use the chromatogram obtained with composition with the following modifications. Use the
reference solution (a) to identify the peaks due to impurity M normalisation procedure.
and bacitracins A, Bl, B2 and B3 (see Figure 0466.-1). Injection Test solution and reference solutions (a),
Relativeretention With reference to bacitracin A (retention (b) and (c).
rime = about 20 min): impurity A = about 0.44; Identification of impurities Use the chromatogram obtained
= =
impurity B about 0.52; impurity C about 0.55; with reference solution (3) to identify the peaks due to
=
bacitracin HI about 0.65; bacitracin H2 =' about 0.67; impurities A, B, C, 1., M, N, 0, P and Q (see Figure
bacitracin B3 = about 0.81; impurity M = about 0.87; 0466.-1); use the chromatogram obtained with reference
= =
impurity N about 0.90; impurity L about 0.93; solution (c) to identify the peaks due to impurities E, F, G,
impurity 0 =
about 1.2; impurities P and Q = about 1.3; and H (see Figure 0466.-2).
=
impurity F = about 1.6; impurity G about 1.8j Limits:
impurity H = about 2.1; impurity E = about 2.8. - sum of impurities Land N: maximum 8.0 per cent;
If necessary, adjust the composition of the mobile phase by - impun°ty E: maximum 4.0 per cent;
changing the amount of organic modifier whilst keeping the - impurity A: maximum 3.5 per cent;
ratio constant between methanol and acetonitrile. - impurities B, M: for each impurity, maximum 3.0 per cent;
System suitability:
- impurity C: maximum 2.5 per cent;
- peak-t~ney ratio: minimum 1.2, where Hp = height - sum of impurities 0, P and Q: maximum 2.5 per cent;
- sum of impun"ties F and G: maximum 2.0 per cent;
above the baseline of the peak due to bacitracin B2 and
H; =height above the baseline of the lowest point of the - impurity H: maximum 1.0 per cent;
curve separating this peak from the peak due to bacitracin - atry otherimpun°ty: for each impurity, maximum
B 1 in the chromatogram obtained with reference 2.0 per cent;
solution (a); - wta/: maximum 23.0 per cent;
- peak-to-vaUey ratio: minimum 1.1, where Hp height= - reporting, threshold: 0.25 per cent.
above the baseline of the peak due to impurity M and Zinc
H; = height above the baseline of the lowest point of the 3.5 per cent to 5.5 per cent (dried substance).
curve separating this peak from the peak due to bacitracin Dissolve 00200 g in a mixture of 2.5 mL of dilute acetic acidR
B3 in the chromatogram obtained with reference and 2.5 mL of water. Add 50 mL of water R, 50 mg of
solution (a); xylenol orange triturate R and sufficient
- signal-to-noise rauo: minimum 50 for the peak due to hexamethylenetetramine R to produce a red colour. Add 2 g of
bacitracin A in the chromatogram obtained with reference
solution (b).
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1-252 Bacitracin Zinc 2022
hexamethylenetetramine R in excess. Titrate with 0.01 1\'1

H2~vX'
sodium edetate until a yellow colour is obtained.
1 mL of 0.01 M sodium edetate is equivalent to 0.654 mg of
Hi CH 3
Zn. S "N
l-l··rl.LeU-o-GIu-L-val-l.Lys-o-orn-l-VaI-o-PheJ
Maximum 5.0 per cent, determined on 1.000 g by drying in H 0 t-Asn-OoAsp-l-His
'Vacuo at 60°C at a pressure not exceeding 0.1 kPa for 3 h.
ASSAY D. 4,1 o-anhydlO[N-[[(4R)-2-[ (1S}-I-amino-2-methylplOpyl]-
Suspend 50.0 mg in 5 mL of water R, add 0.5 mL of di/uI< 4,5-dihydro-I,3-thiazol-4-yl]catbonyl]-r.-leucyl-D-«-
hydrochloric acidR and dilute to 100.0 mL with waterR. glutamyl-L-valyl-L-lysyl-D-omithyl-L-valyl-D-phenylalanyl-
Allow the solution to stand for 30 min. Carry out the L-histidyl-D-«-aspattyl-L-asparagine] (bacitracin E),
microbiological assay of antibiotics (2.7.2). Use bacitracin
H. CH3
zinc CRS as the chemical reference substance.
STORAGE
In an airtight container. If the substance is sterile, the
0.:rz;CH'
S "N
~l'leu_o-GlU-l_".-l'lys-o-O,"-l"Ie-o-Ph.J
container is also sterile and tamper-evident.
IMPURITIES
Specified impun"ties A, B, C, E, F, GJ H, L, M, N, 0, P, Q. tl-Asn--o-Asp -l-His
Otherdelectable impurities (the following substances would, if
present at a sufficient level, be detected by one or other of the tests E. 4,10-anhydro[N-[[2-[(2S)-2-methyl-l-oxobutyl]-1,3-
in the monograph, They arelimited by the general acceptanu thiazol-4-yl]carbonyl]-L-leucyl-D-«-glutamyl-L-isoleucyl-r.-
rnten'on for other/unspecified impurities. It is therefore not Iysyl-D-omithyl-L-isoleucyl-D-pbenylalanyl-L-histidyl-D-«-
neussary 10 identify these impun'tUs for demonstration of aspartyl-t-esparagine], (bacitracin F),
compliance. See also 5.10. Control of impurities in substances for
phannaceutical use) D, 1, J, K.
H~ CH 3
S
H~CH' N
.
Y.ll-Leu- o-Glu --l-lle- L-Lys-o-Om-L-Val- o-PheJ
H 0 Ll_Asn_o-Asp_l_HiS
A. 4,1 O-anhydro[N-[[(4R)-2-[(IS}-I-amino-2-methylpropyl]- F. 4,1 O-anhydro [N- [[ 2- (2-methyl-l-<>xopropyl)-1,3-thiazol-4-

4,5-<lihydro-1 ,3-thiazol-4-yl]carbonylj-t.-leucyl-n-c- yl]carbonyl]-L-Ieucyl-D-«-glutamyl-L-isoleucyl-L-lysyl-D-
glutamyl-L-isoleucyl-L-Iysyl-D-omithyl-L-valyl-D- omithyl-L-isoleucyl-D-phenylalanyl-L-histidyl-D-«-aspattyl-
phenylalanyl-L-histidyl-D-«-aspartyl-L-asparagine] r-asparagine] (bacitracin Hl),
(bacitracin DI, bacitracin C2),
°XCC~
CH,
~2~y(
----l.. CH
3
S N
~l-l.U_o-GIU-l_lle-l-lYS-o-Om-l-val-o-ph<lJ
S N
~ . rt-Leu-s- o-Glu -l-Val -l-Lys'" c-om-e-r-ue -- o-PheJ
o Ll_Asn_o-Asp_L_HIs t L-Asn -o-Asp --L-His
B. 4,10-anhydlO[N-[[(4R)-2-[(IS}-I-amino-2-methylplOpyl]-
G. 4,1 0-anhydfo[N-[[2-[(2S}-2-methyl-I-oxobutyl]-1,3-
4,5 -dihydro-I ,3- thiazol-4-yl]carbonyl]-L-Ieucyl-D-«- thiazol-4-yl]carbonyl]-L-leucyl-D-«-glutamyl-L-isoleucyl-L-
glutamyl-L-valyl-L-Iysyl-D-omithyl-L-isoleucyl-D- lysyl-D-omithyl-L-valyl-D-phenylalanyl-L-histidyl-D-«-
phenylalanyl-L-histidyl-D-«-aspartyl-L-asparagine]
aspartyl-L-asparagine] (bacitracin H2),
(bacitracin D2, bacitracin C3),
H. CH3
0.:rz;c~
S N
~l-l.U_o-GIU-l-VaJ -l-Lys - o-Orn-l-Ile -o-PheJ
ll-Asn -o-AsP--l-His
C. 4,1 O-anhydlO[N-[[ (4R)-2-[ (IS,2S}-I-amino-2-

methylbutyl] -4,5-dihydlO-1 ,3-thiazol-4-yl] carbonylj-r- H.4,10-anhydro[N-[[2-[(2S}-2-methyl-I-oxobutyl]-1,3-
leucyl-D-«-glutamyl-L-valyl-L-Iysyl-D-omithyl-L-valyl-D- thiazol-4-yl]carbonyl]-L-Ieucyl-D-«-glutamyl-L-valyl-L-Iysyl-
phenylalanyl-L-histidyl-D-«-aspartyl-L-asparag;ne] D-omithyl-L-isoleucyl-D-phenylalanyl-L-histidyl-D-«-
(bacitracin D3, bacitracin Cla), aspartyl-L-asparagine] (bacitracin H3),
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CH,
arCH,
s "'N
~'-L'U-O-GJU-'."'-'_Lys-o-arn-'-V"-O-Ph.J
o t Asn --o-Asp ......
L- l-Hls
N. 4, I O-anhydro[N-[[(4S)-2-[(lS,2S)-I-amino-2-
1. 4, I Q--anhydro[N-[[2-(2-methyl-I-oxopropyl)-1,3-thiazol-4- methylbutyl]-4,5-<1ihydro-l,3-thiazol-4-yl]carbonyl)-L-
yl]carbonyl)-L-Ieucyl-o-<l-gluramyl-L-isoleucyl-L-Iysyl-o- leucyl-o-<l-gluramyl-L-isoleucyl-L-lysyl-o-omithyl-L-
omithyl-L-valyl-o-phenylalanyl-L-histidyl-o-<l-aspartyl-L- isoleucyl-o-phenylalanyl-L-histidyl-o-<l-aspartyl-L-
asparagine) (bacitracin II), asparagine] (bacitracin Z),
CH,
arCH,
s "'N
~'-L'U-O-GJU-'-V"-'_Lys-o-a,"-'-".-O-Ph.J
a t ·
l-Asrl-o-Asp-L.HIS
o. Mil = 5-methylene-L-isoleucine: 4,IO-anhydro[N-[[(4R)-2-
1. 4, I O-anhydro [N- [[2-(2-methyl-I-oxopropyl)-1,3-thiazol-4- [(I S,2S)-1-amino-2-methylbutyl]-4,5-dihydro-l ,3-thiazol-
yl]carbonyl)-L-Ieucyl-o-<l-gluramyl-L-valyl-L-Iysyl-o- 4-yl)carbonyl)-L-leucyl-o-<l-glutarnyl-5-methylene-L-
omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-o-«-aspartyl- isoleucyl-L-Iysyl-o-omithyl-L-isoleucyl-o-phenylalanyl-L-
L-asparagine] (bacitracin 12), histidyl-n-c-asperryl-r-asparagine] (bacitracin JI),
aXCC~ ~XCC~
s "'N S "'N
~'_Leu-o-GIU-'_V'I-'_Lys_o-arn-'-V,,-o-PheJ L ~
T·l
L-leU-o-GIU-L-ue-L-lys-o-am--Mil-o-Ph9 J
H 0 ll.Asn---o-Asp-l-HIS
o tL-Asn'-'O-AsP-L-His
K. 4, I O-anhydro [N- [[2-[(2S)-2-methyl-I-oxobutyl)-1,3-

=
P. Mil 5-methylene-L-isoleucine: 4,IO-anhydro[N-[[(4R)-2-
[(I S,2S)-1-amino-2-methylbutyl)-4,5 -dihydrc-I,3-thiazol-
thiazol-4-yl)carbonyl]-L-Ieucyl-o-<l-g1uramyl-L-valyl-L-lysyl- 4-yl)carbonyIJ-L-leucyl-o-<l-glutarnyl-L-isoleucyl-L-lysyl-o-
o-omithyl-L-valyl-o-phenylalanyl-L-histidyl-o-<l-aspartyl-L- omithyl-5-methylene-L-isoleucyl-o-phenylalanyl-L-histidyl-
asparagine] (bacitracin 13), n-c-asparryl-t-asparagine] (bacitracin J2),
H2X
Hi H~ H, CH3
s "'N
c~
H~C~
y" loLeU-o-GIu-L-lIe-L-Lys-o-om-L-lle-o-Phe J S "'N
y .~ t-Leu-e-o-Glu- L-Ile'" L-lys ..... o-Om-e- L·lle -- D-PheJ
H ~ LL-Asn-o-Asp-L-His
H 0 lLoAsn-o-Asp-L-His
L. 4, I Q--anhydro [N-[[(4R)-2-[ (IR,2S)-I-amino-2-

Q. 4, I O-anhydro[N-[[(4R)-2-[(IS,2S)-I-amino-2-methylpent-
methylbutyl]-4,5-dihydro-1 ,3-thiazol-4-yl]carbonyl]-L-
4-en-I-yl)-4,5-dihydro-l, 3-thiazol-4-yIJ carbonyl)-L-leucyl-
leucyl-o-<l-g1utamyl-L-isoleucyl-L-lysyl-o-omithyl-L-
D-<l-glutamyl-L-isoleucyl-L-Iysyl-D-omithyl-L-isoleucyl-o-
isoleucyl-o-phenylalanyl-L-histidyl-o-«-aspartyl-L-
phenylalanyl-L-histidyl-o-<l-aspartyl-L-asparagine)
asparagine] (bacitracin X),
(bacitracin J3).
___________________ "'E"
M.4,IO-anhydro[N-[[2-[(IS,2S)-I-amino-2-methylbutyl]-1,3-
thiazol-4-yl)carbonyl]-L-leucyl-o-<l-g1uramyl-L-isoleucyl-L-
Iysyl-o-omithyl-L-isoleucyl-o-phenylalanyl-L-histidyl-o-«-
aspartyl-L-asparagine] (bacitracin Y),
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1-254 Baclofen 2022
Reference solution Dissolve 10 mg of badofen CRS in the

Baclofen mobile phase and dilute £0 10 mL with the mobile phase.
(Ph. Ellr. monograph 0653) Pia,. TLC silica gel G plau R.
Mobile phase anhydrous lonnie acid R, water R, methanol R,
Nil, chloroform R, erhyl aceta,. R (5:5:20:30:40 VIVIVIVIV).
( H
App/kation 5 ~L.
0~H andenantiomer
Development Overa path of 12 em.
CI N Drying Allow the solvents to evaporate.
Daeaion Spray with ninhydrin solution Rl until the plate is
C IOH12CINO, 213.7 1134-47-0 slightly wet. Place in an oven maintained at 100 °C for
10 min. Examine in daylight.
Acdon and use Resvlu The principal spot in the chromatogram obtained
Skeletal muscle relaxant. with the test solution is similar in position, colour and size to
Preparations the principal spot in the chromatogram obtained with the
Baclofen Oral Solution reference solution.
Baelofen Tablets TESTS
PhE" _ Appearance of soludon
The solutionis not more opalescent than reference
DEFINITION suspension II (2.2.1) and not more intensely coloured than
(3RS)-4-Amino-3-(4-chlorophenyl)butanoic acid. reference solution BY, (2.2.2, MetIwd II).
Content Dissolve 0.50 g in 1 M sodium hydroxide and dilute to 25 mL
98.0 per cent to 101.0 per cent (anhydrous substance). with the same solvent.
Appearance liquid chromatography (2.2.29).
White or almost white powder. Testsolution Dissolve 25.0 mg of the substance to be
Solubility examined in the mobile phase and dilute to 10.0 mL with
Slightly soluble in water, very slightly soluble in ethanol the mobile phase.
(96 per cent), practically insoluble in acetone. It dissolves in Reference soilltion (a) Dissolve 25.0 mg of baelofen
dilute mineral acids and in"dilute solutions of alkali impwity A CRS in the mobile phase and dilute to 10.0 mL
hydroxides. with the mobile phase.
It shows polymorphism (5.9). Reference solution (b) Dilute 1.0 mL of reference solution (a)
IDENTIFICATION
Firstidentification: B. Reference soilltion (c) Dilute 2.0 mL of the test solution 10
Reference solllrion (d) Dilute 2.0 mL of the test solution and
2.0 mL of reference solution (a) to 100.0 mL with the
(2.2.25).
mobile phase.
Test solution Dissolve 70 mg in water R and dilute to
Column:
J00.0 mL with the same solvent.
- size: 1= 0.25 m, 0 = 4.0 mm;
Spe<tral range 220-320 nrn, - stationary phase: octadecy/si/yl silka gelfor chromatography R
Absorption maxima At 259 nm, 266 nm and 275 nm. (10 urn).
Resolution (2.2.25): minimum 1.5 for the absorbance ratio. Mobile phase Dissolve 1.822 g of sodium hexanesulfomue R in
Spedfic absorbance at the absorption maxima: I L of a mixture of 560 volumes of water R, 440 volumes of
- at 259 run: 9.8 to 10.8; methanol Rand 5 volumes of glacial acetic add R.
- at 266 run: 11.5 to 12.7; Flow ra,. 2.0 mUmin.
- at 275 nm: 8.4 to 9.3. Deuction Spectrophotometer at 266 om.
B. Infrared absorption spectrophotometry (2.2.24). Injection 20 llL of the test solution and reference
Preparation Discs prepared using 3 mg of substance and solutions (b), (c) and (d).
300 mg of potassium bromide R. Run lime 5 times the retention time of baclofen.
Comparison baelofen CRS. System suitability Reference solution (d):
If the spectra obtainedin the solid state show differences, - resolution: minimum 2.0 between the peaksdue to
dissolve 0.1 g of each of the substances separately in 1 rnL of baelofen and impurity A.
dilll" sodium hydroxide solution R and add 10 mL of ethanol Limits:
(96 per cen!! Rand 1 mL of dilll,. acetic acidR. Allow to - impurity A: not more than the area of the principal peak
stand for 1 h. Filter, wash the precipitate with ethanol in me chromatogram obtained with reference solution (b)
(96 per cent) R and dry in vacuo. Prepare new discs and (1.0 per cent);
record the spectra. - total: not more than the area of the principal peak in the
C. Thin-layer chromatography (2.2.27). chromatogram obtainedwith reference solution (c)
Test solution Dissolve 10 mg of the substance to be (2.0 per cent).
examined in the mobile phase and dilute to 10 rnL with the Water (2.5.12)
mobile phase. Maximum 1.0 per cent, determined on 1.000 g.
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2022 Bambuterol Hydrochloride 1-255
Sulfated ash (2.4./4) Solubility

Maximum 0.1 per cent, determined on 1.0 g. Freely soluble in water) soluble in ethanol (96 per cent»
ASSAY
practically insoluble in heptane.
Dissolve 0.1500 g in 50 mL of anhydrous acetic acid R. I, shows polymorphism (5.9).
Titrate with 0.1 M percblonc (Kid, determining the end-point IDENTIFICATION
potentiometrically (2.2.20). A. Infrared absorption spectrophotometry (2.2.24).
I mL of 0./ M perchloric acid is equivalent to 21.37 mg Comparison bombutero; hydrochlonile CRS.
of C,oH 12CIN02 • If me spectra obtained show differences) dissolve the
IMPURITIES substance to be examined and thereference substance
Specified impun',ies A. separately in a mixture of 1 volume of water Rand 6 volumes
Other deteaabie impurities (the fo//awing subslances would, if of outone R) cool in ice to precipitate and dry both
present at a sufficient level, be detected by oneor other of the tests precipitates in vacuo at 50°C to constant weight. Record new
in the monograph. They are limited by the general acceptance spectra using the residues.
criterion for other/unspecified impurities and/or by the general B. Ir gives reaction (a) of chlorides (2.3./).
monograph Subs'ances for pharmaceutical use (2034). It is TESTS
therefore not neussary 10 Mcntify these impun"ties for Solution S
demonstration of compliance. See also 5. JO. Control of impurities Dissolve 4.0 g in carbon dioxide-free water R and dilute to
in substances for pharmaceutical use) B. 20.0 mL with the same solvent.
~
.?
0 To 10 mL of solution S add 0.2 mL of methyl red solution R
~ and enantiomer and 0.2 mL of 0.0/ M hydroch/ori< acid. The solution is red.
Add 0.4 mL of 0.0/ M sodium hydroxide. The solution is
CI~ yellow.
A. (4RS)-4-(4-chlorophenyl)pyrrolidin-2-one, -n.IO' '0
+ 0.10'.
Dilute I mL of solution S to 10 mL with carbon dioxide-free
o
',!:.NH,
H
water R.
Related substances
co H and enanllomer
if
o?' '
I 0.050 M phosphate buffersolution Dissolve 6.90 g of sodium
CI '" dihydrogen phosphate monohydrate R in waterfor
'0
chromawgraphy R, adjust pH 3.0 with a 50 gIL solution of
B. (3RSJ-5-amino-3-( 4-chlorophenyl)-5-oxopentanoic acid. dilute phosphonc acid Rand dilute to 1000 mL with waterfor
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r chromawgraphy R.
Test solution Dissolve5.0 mg of the substance to be
the mobilephase.
Bambuterol Hydrochloride Reference solution (a) Dissolve the contents of a vial of
bambuterol impurity mixtureCRS (impurities C and D) in
(ph. Bur. monograph /293) I mL of the mobile phase.
Reference sdution (b)Dilute 1.0 mL of the test solution '0
solution '0
10.0 mL with the 'mobile phase.
HCI Co/umn:
- size: I:::: 0.15 m, 0 = 4.6 mm;
- stationary phase: base-deactivated end-capped otladecy/si/yl
silica gelfor chromawgraphy R (5 urn).
jWobile phase Dissolve 1.3 g of sodium octanesulfonate R in
403.9 8/732-46-9 430 mL of a mixture of 25 volumes of aeewrtitrj/e R/ and
75 volumes of methanol R2J then mix the solution with
Action and use 570 mL of the 0.050 M phosphate buffer solution.
Betaj-adrenoceptor agonist; bronchodilator. Flew rate 1.5 mUmin.
PhE" _ Detection Spectrophotometer at 214 am.
/njC<lion 20 ~L; inject the mobile phase as a blank.
DEFINITION
5-[(I RSJ -2-( terl-Butylamino)-I-hydroxyethy1]-1,3-phenylene Run time 1.5 times the retention time of bambuterol.
bisfdimethylcarbamate) hydrochloride. Identi/icau"on of impurities Use the chromatogram supplied
with baminuerol impurity mixture CRS and the chromatogram
Content
98.5 per cent to 101.5 per cent (anhydrous substance). obtained with reference solution (a) to identify the peaks due
to impurities C and D.
CHARACTERS
Appearance
White or almost white) crystalline powder.
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1-256 Barbital 2022
Relativeretention Wilh reference to bambuterol (retention

%H' 0ifHPH
impurity D = about 050.
H,C' Y ""
I CH,
o CH ~ andenantiomer
SystemsuitabIlity Reference solution (a): , a
- resolution: minimum 2.0 between the peaks due to ,N-y-0
H3C II
impurities C and D. o
Calculation of percemage contents:
- for each impurity, use the concentration of bambuterol D. 5-[(lRS)-I-hydroxyethyl]-1,3-phenylene bis
hydrochloride in reference solution (b). (dimethylcarbamate),
Limits:
CH, 0
- impurity C: maximwn 0.2 per cent;
- unspecified impuntie5: for each impurity, maximum H , c ) y o v iGH,
0.10 per cent;
o CH,"-
- wtal: maximwn 0.4 per cent; ,
- reporting threshold: 0.05 per cent. H,C,NyO
Water (2.5.12) o
Maximwn 0.5 per cent, determined on 0.500 g.
Sulfated ash (2.4.14) E. 5-acetyl-I,3-phenylene bis(dimethylcarbamate),
ASSAY
Dissolve 0.320 g in 50 mL of ethanol (96 perun!> R and add
5 mL of 0.01 M hydrochlon', acid. Carry out a potentiometric
titration (2.2.20), using 0.1 M sodium hydroxide. Read
the volume added between the 2 points of inflexion.
ClsH30ClN30j.
F. 5-[ (rer-butylamino)acetyl]-l ,3-phenylene bis
IMPURITIES (dimethylcarbamate).
Specified impurities C. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw
Otherdececrable impurities (thefollowing substances would, if
present at a sufficient leveJ~ be detected by oneor other of the tests
m the monograph. They are limiud by the general acceptance
en«rioofor other/unspecified impurities and/or by the general
monograph Substances for pharmaceutical use (2034). II ~
Barbital
therefore not necmary to identify these impuniies for
demonstration of compliance. See also 5.10. CoolT01 of impurilles
in substances for pharmaceutical use) A, B, D, H, F.
H OH
H O T " ~,/CH,
/ \ andenantlomer
~ H3C CH,
OH 184.2 57-44-3
A. 5-[(1RS)-2-(tert-butyl.mino)-I-hydsoxyethyl]benzene-I,3- Action and use

diol (terbutallne), Barbiturate.
H PhEw _ _~ ~ - _ - _ - - - - - - - - - - -
'r , i f HOH'OH
,N-y-0 "" DEFINITION
H,C II I Barbital contains not less than 99.0 per cent and not more
andenantiomer
o CH
, a """" than the equivalent of 101.0 per cent of
,N-y-0 5,5-diethylpyrimidine-2,4,6(IH,3H,5H)-trlone, calculated
H3C II
with reference to the driedsubstance.
o
CHARACTERS
B. 5-[(IRS)-1,2-dihydroxyethyl]-1,3-phenylene bis A white or almost white, crystalline powder or colourless
(dimethylcarbamate), crystals, slightly soluble in water, soluble in boiling water and
in alcohol. It forms water-soluble compounds with alkali
CH:J H OH hydroxides and carbonates and with ammonia.
H,c'~yO~~XCH'
o Y H,C CH,
andenantiomer IDENTIFICATION
Fi~ti~t~cariOn:A, B.
OH
A. Determine the melting point (2.2.14) of the substance to
C. 3-[(IRS)-2-(tert-butylamino)-I-hydsoxyethyl]-5- be examined. Mix equal parts of the substance to be
hydroxyphenyl dlmerhylcarbamate, examined and barbital CRS and determine the melting point
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2022 Barium Sulfate 1-257
of the mixture. The difference between the melting points ASSAY

(which are about 190 °C) is not greater than 2 "C. Dissolve 85.0 mg in 5 mL of pyridine R. Add 0.5 mL of
R Examine by infrared absorption spectrophotometry thymolphlhalein solution R and 10 mL of silver nitrate solution
(2.2.24» comparing with the spectrum obtained with in pyridine R. Titrate with 0.1 M ethandicsodium hydroxide
barbital CRS. until a pure blue colour is obtained. Carry out a blank
C. Examine by thin-layer chromatography (2.2.27), using titration.
silica gd GF254 R as the coating substance. 1 mL of 0.1 M emanolic sodium hydroxide is equivalent to
Test solution Dissolve 75 mg of the substance to he 9.21 mg of C.H 12NzO,.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
examined in alrohol R and dilute to 25 mL with the same
solvent.
Reference solution Dissolve 75 mg of barbital CRS in
alcohol R and dilute to 25 mL with the same solvent.
Apply separately to the plate 10 ~L of each solution. Develop Barium Sulfate
over a path of 18 em using me lower layer of a mixture of
5 volumes of concentrated ammonia R, 15 volumes of alcohol R
and 80 volumes of chloroform R. Examine immediately in BaSO, 233.4 7727-43-7
ultraviolet light at 254 run. The principal spot in the
chromatogram obtained with the test solution is similar in Action and use
position and size to the principal spot in the chromatogram Radio-opaque substance used ln. the investigation of the
obtained with the reference solution. gastro-intestinal tract.
D. It gives the reaction of non-nitrogen substituted Preparation
barbiturates (2.3.1). Barium Sulfate for Suspension
TESTS PhE" _
CHARACTERS
Dissolve 1.0 g in a mixture of 4 mL of dilute sodium hydroxide
Appearance
solution Rand 6 mL of water R. The solution is clear (2.2.1)
Fine, white or almost white powder, free from gritty particles.
and not more intensely coloured than reference solution YI)
(2.2.2, MethodII). Solubility
Practically insoluble in water and in organic solvents. It is
Acidity
very slightly soluble in acids and in solutions of alkali
Boil 1.0 g with 50 mL of water R for 2 min, allow to cool
hydroxides.
and filter. To 10 mL of the filtrate add 0.15 mL of methylred
solution R. The solution is orange-yellow. Not more than IDENTIFICATION
0.1 mL of 0.1 M sodium hydroxide is required to produce a A. Boil a suspension of 0.2 g with 5 mL of a 500 gIL
pure yellow colour. solution of sodium carbonate R for 5 min, add 10 mL of
Related substances water RJ filter and acidify a pan of the filtrate with dilute
Examine by thin-layer chromatography (2.2.27), using silica hydrochloric add R. The solution gives the reactions of sulfates
gel GFZ54 R as the coating substance. (2.3.1).
Test solution Dissolve 1.0 g of the substance to be examined B. Wash the residue collected in the preceding test with
in akohol R and dilute to 100 mL with the same solvent. 3 successive small quantities of water R. To the residue add
5 mL of dilute hydrochloric acid R, filter and add to the filtrate
Reference solution Dilute 0.5 mL of the test solution to
0.3 mL of d,lute sulfun·c acid R. A white precipitate is formed
100 mL with alcohol R.
that is insoluble in dilure sodium hydroxide solutitm R.
Apply separately to the plate 20 ~L of each solution. Develop
over a path of 15 cm using the lower layer of a mixture of TESTS
5 volumes of concentrated ammonia R, 15 volumes of akohol R Solution S
and 80 volumes of chloroform R. Examine immediately in To 20.0 g add 40 mL of distilled waler Rand 60 mL of dl/ute
ultraviolet light at 254 nm. Spray with diphenykarbazone acetic acid R. Boil for 5 min, filter and dilute the cooled
mercuric reagent R. Allow the plate to dry in air and spray filtrate to 100 mL with distilled waterR.
with freshly prepared alcoholic potassium hydroxide solutUm R Acidity or a1kallnity
diluted 1 in 5 with a/dehyde-jree alrohol R. Heat at 100 'C to Heat 5.0 g with 20 mL of carbon dioxide-free waterR on a
105 °C for 5 min and examine immediately. When examined water-bam for 5 min and filter. To 10 mL of the filtrate add
in ultraviolet light and after spraying, any spot in the 0.05 mL of bromomymd bluesolution Rl. Not more than
chromatogram obtained with the test solution, apart from the 0.5 mL of 0.01 M hydrochloric acid or 0.01 M sodium
principal spot, is not more intense man the spot in the hydroxide is required to change the colour of the indicator.
chromatogram obtained with the reference solution Acid-soluble substances
(0.5 per cent). Maximum 0.3 per cent.
Loss on drying (2.2.32) Evaporate 25 mL of solution S to dryness on a water-bath
Not more than 0.5 per cent, determined on 1.00 g by drying and dry to constant mass at lOa-105°C. The residue weighs
in an oven at 105 °C. a maximum of 15 mg.
Sulfated ash (2.4.1 if) Oxidisable sulfur compounds
Not more than 0.1 per cent, determined on 1.0 g. Shake 1.0 g with 5 mL of mater R for 30 s and filter. To the
filtrate add 0.1 mL of starch solution R, dissolve 0.1 g of
potassium iodide R in the mixture, add 1.0 mL of a freshly
prepared 3.6 mgIL solution of potassium iodate Rand 1 mL
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1-258 Barium Sulfate for Suspension 2022
of 1 M hydrochloric acid and shake well. The colour of the potassium carbonate sesquihydrate and mix. Heat to 1000° and
solution is more intense than that of a standard prepared at maintain at this temperature for 15 minutes. Allow to cool
the same time and in the same manner, but omitting the and suspend the residue in 150 mL of water. Wash the dish
potassiwn iodate. with 2 mL of 6M acetic acid and add the washings to the
Soluble barium salts suspension. Cool in ice and decant the supernatant liquid,
Maximum 10 ppm. transferring as little of the solid matter as possible to the
filter. Wash the residue with successive quantities of a
To 2.5 mL of a 0.2 mg/L solution of ban'um nitrate R in a
2% w/v solution of sodium carbonate until the washings are
mixture of 30 volumes of ethanol (96 per emu Rand
free from sulfate and discasd the washings. Add 5 mL of 2M
70 volumes of water R, add 10 mL of dilute sulfuric acid R.
hydrochloric. add to the filter, wash through into the vessel
Shake and allow to stand for 5 min. To 1 mL of Ihis solution
containing the bulk of the solid matter with water, add 5 mL
add 10 mL of solution S. Prepare a standard in the same
of hydrochloric acid and dilute to 100 mL with water.
manner using 10 mL of barium standard solution (2 ppm
Add 10 mL of a 40% w/v solution of ammonium acetate,
BaJ R instead of solution S.
25 mL ofa 10% w/v solution of potassium d~hromate and
After 10 min, any opalescence in the test solution is not 10 g of urea. Cover and digest in a hot-air oven at 80 0 to 85°
more intense than chat in the standard. for 16 hours. Filter whilst still hot through a sintered-glass
Loss on ignition filter (ISO 4793, porosity grade 4, is suitable), washing the
Maximwn 2.0 per cent, determined on 1.0 g at precipitate initially with a 05% wlv solution of potassium
600 ± 50 "C. dichromate and finally with 2 mL of water. Dry to constant
_____________________ "',<1 weight at 105°. Each g of the residue is equivalent to
0.9213 g of barium sulfate, BaSO,.
Barium Sulfate for Suspension

Barium Sulphate for Suspension
Beclometasone Dipropionate
Action and use Anhydrous Beclometasone Dipropionate
Radio-opaque preparation used in the investigation of the (Ph. Bur. monograph 0654)
gastro-intestinal tract.
Preparation
Barium Sulfate Oral Suspension
DEFINITION
Barium Sulfate for Suspension is a dry mixture of Barium
Sulfate with a suitable dispersing agent and may contain
suitable flavours and suitable antimicrobial preservatives.
Content ofbariwn sulfate, BaSO"
90.0 to 110.0% of the stated amount.
Cu.H"CIO, 521.0 5534-09-8
CHARACTERISTICS
A fine, white or creamy white powder. Action and use
Glucocorticoid.
IDENTIFICATION
A. Ignite 1 g to constant weight. To 0.2 g of the residue add Preparations
5 mL of a 50% wlv solution of sodium carbonate and boil for Beclometasone Cream
5 minutes. Add 10 mL of water and filter. Reserve the Beclometasone Aqueous Nasal Spray
residue for test B. Acidify a portion of the filtrate with Beclometasone Inhalation Powder
2M hydrochloric acid. The solution yields the reactions Beclomerasone Inhalation Powder, pre-metered
characteristic of sulfates, Appendix VI.
Beclometasone Ointment
E. Wash the residue reserved in test A with water, add 5 mL
Beclometasone Pressurised Inhalation
of 2M hydrochloric acid, mix well and filter. Add 0.3 mL of 1M
sulfu,* acid to the filtrate. A white precipitate is produced
which is insoluble in 2M hydrochlori< acid.
"''''------------------
DEFINITION
TESTS 9-Chloro-11 P-hydroxy-16Il-methyl-3,20-dioxopregna-I,4-
Acidity or alkalinity diene-17,ZI-diyl dipropanoate.
pH of an aqueous suspension containing the equivalent of
Content
60% w/w of Barium Sulfate or, for lower strengths, the
aqueous suspension at the strength of intended use, 3.5 to
8.5, Appendix V L. CHARACTERS
Loss on drying Appearance
When dried at 105 0 for 4 hours, loses not more than 1.0% of White or almost white, crystalline powder.
its weight. Use I g. Solubility
ASSAY
sparingly soluble in ethanol (96 per cent).
To a quantity containing 0.6 g of Barium Sulfate in a
platinum dish add 5 g of sodium carbonate and 5 g of IDENTIFICATION
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2022 Beclometasone Dipropionate 1-259
Comparison anhydrous beclometasone dipropionate CRS. dipropionate for system suitability CRS and the chromatogram
B. Treat 25 mg by the oxygen-flask method (2.5.1a). Use a obtained with reference solution (b) to identify the peakdue
mixture of I mL of I M sodium hydroxide and 20 mL of to impurity D.
water R to absorb the combustionproducts. The solution Relative retention Withreference to beclometasone
gives reaction (a) of chlorides (2.1. I). dipropionate (retention time == about 25 min):
C. Loss on drying (see Tests). impurity A == about 0.3; impurity B == about 0.6;
impurity D == about 1.1; impurity M == about 1.2;
TESTS impurity L == about 1.3;impurity C == about 1.8;
Specific optical rotation (2.2.7) =
impurity N about 2.0; impurity F about 2.2.=
+ 108 to + 115 (dried substance). System suitability Reference solution (b):
Dissolve 0.100 g in ethauol (96 perceut) R and dilute to - peak-to-1Jal/ey ratio: minimwn 1.5, where HI' == height
10.0 mL with the same solvent. above the baseline of the peak due to impurity D and
Related substances H; == height above the baselineof the lowest point of the
Liquid chromatography (2.2.29). curve separating this peak from the peak due to
Solventmixture Mobile phase A, mobile phase B bedometasone dipropionate.
(45:55 VIl'). Limits:
Test solution (a) Dissolve 50.0 mg of the substance to he - correction factors: for the calculation of content, multiply
examined in 28 mL of mobile phase B and dilute to 50.0 mL the peak areas of the following impurities by the
with mobile phase A. corresponding correction factor: impurity F == 1.3;
Tes, solutiou (1)) Dilute 1.0 mL of test solution (a) to impurity M == 2.0;
- impurity L: not more than 6 times the area of the principal
Reference solution (a) Dilute 5.0 mL of test solution (b) to solution (a) (0.6 per cent);
100.0 mL with the solvent mixture. - impurities B, F, M: for each impurity, not more than
Reference solution (b) Dissolve 5 mg of bedometasone 5 times the area of the principal peak in the
dipropiouatefar system suitabili(Y CRS (containing impurity D) chromatogram obtained with reference solution (a)
in 3 mL of mobile phase B and dilute (0 5 mL with mobile (0.5 per cent);
phase A. - impurities A, D, N: for each impurity, not more than twice
Reference solutiou (c) Dissolve 5 mg of beclometasone the area of the principal peak in the chromatogram
dipropionate for peak identification CRS (containing impurities obtained with reference solution (a) (0.2 per cent);
A, B, C, Land M) in 3 mL of mobile phase B and dilute to - impun"ty C: not more than 1.5 times the area of the
5 mL with mobile phase A. Use 1 mL of this solution to principal peak in me chromatogram obtained with
dissolve the contents of a vial of bedometasone dipropiqnau reference solution (a) (0.15 per cent);
impurities F aud N CRS. - unspecified impurities: for each impurity, not more than the
Reference solution (d) Dissolve 50.0 mg of anhydrous area of the principal peak in the chromatogram obtained
bedometasone dipropionate CRS in 28 mL of mobile phase B with reference solution (a) (0.10 per cent);
and dilute to 50.0 mL with mobile phase A. Dilute 1.0 mL - total: not more than 15 times the area of the principal
of this solution to 50.0 mL with the solvent mixture" peak in me chromatogram obtained with reference
Column:
- size: 1== 0.25 rn, 0 == 4.6 mrn;
- stationary phase: spherical difunctionel bonded end-capped
(0.05 per cent).
oCladecyisilyl silica gelfor chromategraphy R (5 urn);
- temperature: 50°C. Loss on drying (2.2.32)
Mobile phase: Maximwn 0.5 per cent, determined on 1.000 g by drying in
- mobile phaseA: 2.72 gIL solution of potassium dihydroge1l an oven at 105 DC for 3 h.
phosphate R adjusted to pH 2.35 with phosphoric acidR; ASSAY
- mobile phase B: tetrahydrojUran R, acetonitrile R, methanol R liquid chromatography (2.2.29) as described in the test for
(5:23:25 VIVIV); related substances with the following modification"
Injection Test solution (b) and reference solution (d).
(min) (per cent VIV) (per cent V/I1 Calculate the percentage content of C2sH37CI07 from the
60
declared content of anhydrous beclometasone dipropionate CRS.
0-4 40
4 - 12 40 --> 45 60 --> 55 IMPURITIES
12 - 59 45 55 Specrfied impurities A, B, C, D, F, L, lW, N.
Otherdetectable impun',ies (thefollowiug substances would, if
Flow rate 1.4 mUmin. present at a sufficient level, be detected by one or other of the tests
Detection Spectrophotometer at 254 run. in the monograph. Theyare limited by the general acceptance
Injection 20 ul of test solution (a) and reference
oiteiion for other/unspecified impurities ondlor by the general
monograph Substauces for pharmaceutical use (2034). I, is
therefore not necessary tQ identify these impuniies for
Identification of impurities Use the chromatogram supplied demonstration of compliance. See also 5.10. Control of impurities
with beclometasone dipropionale for peak identification CRS and in substances for pharmaceutical use) E, H, I, J, 0, Q, R, S,
the chromatogram obtained with reference solution (c) to u,v.
identify the peaks due to impurities A, B, C, F, L, M and N;
use the chromatogram supplied with bedometasone
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2022
1-260 Beclometasone Dipropionate
o '.
H B'
A. 9-chloro-11 p, 17-dihydroxy-Ieji-methyl-3,20-dioxopregna-
F. 6~_bromo_9_chloro_llll-hydroxy_16Il-methyl-3,20-
1,4-dien-21-yl propanoate (becJometasone 21-propionate),
dioxopregna-l,4-diene-17,21-diyldipropanoate,
o
H. c-chloro-t t p,21-dihydroxy-16p-methyl-3,2o-dioxopregna-
B. 21-(acetyloxy)-9-chloro-11 P_hydroxy_16p_methyl_3,20_ 1,4-dien-17-yl propancate (beclometasone 17-propionate),
dioxopregna-l,4-dien-17-yl propanoate (beclometasone
21-acetate 17-propionate),
o
o I. 16p_methyl_3,20_dioxopregna_I,4,9(l1)_triene_17,21-diyl
dipropanoate,
C. 9-chlo[<>-11 p-hydroxy-16Il-methyl-3,2o-diox<>-17-
(propanoyloxy)-pregna-I,4-dien-21-yl butanoate o
(beclometasone 21-butyrate 17-propionate),
°O~CH3
o
o~CH3
°O~CH3
»<: -"....
\ CH3
''''/'"--,,,,0
~CH3 H
-, CH3 o
H
J. 9, II p_epoxy_16p_methyl_3,20_diox<>-9Il-pregna-I,4-diene-
o 17,21-diyl diprcpanoate,
D, 9_bromo_llll-hydroxy_16p_methyl_3,20_dioxopregna_I,4_
diene-I? ,21-diyl dipropanoate,
o
°O~CH3
,--,IA-·-·O
~CH3
, CH3
'H
L. 9-<:hloro_llll-hydroxy-16Il-methyl-3,20-dioxopregn-4-ene-
o 17,21-diyl dipropanoate,
H \~I
E. 6~,9_dichlo[o_llll-hydroxy_16p_methyl_3,20_dioxopregna
1,4-diene-17,21-diyl dipropanoate,
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2022 Beclometasone Dipropionate Monohydrate 1-261
o o
o °o~CH3 °O~CH3
.... O~CH3 '~i.~.L"{"_'O~C~
'. CH3
H
o o
M. 9-chioro-ll p-hydroxy-16Il-methyl-3,2o-dioxopregna-4,6- S. 9-chioro-16p-methyl-3,20-dioxopregna-1 ,4-diene-

diene-17J21-diyl dipropanoate, I Ip, 17,21-triyl tripropanoate (beclometasone
tripropionate),
OH
o
o~CH3
....
'. CH3
B' H
o o
N. 2-bromo-9-chioro-11 p-hydroxy-16Il-methyl-3,2o- U. 9, II p-epoxy-21-hydroxy-16Il-methyl-3,20-dioxo-9P-

dioxopregna-l A-diene-I?,21-diyl dipropanoate, pregna-I ,4-dien-I? -yl propanoate,
o
o °O~CH3 o
"'-/~,I _-<.... O~CH3

CH,
H
O. 9, II p-dichioro-16p-methyl-3,20-dioxopregna-I,4-diene- V. 9,IIfl-epoxy-17-hydroxy-16p-methyl-3,2o-dioxo-9P-
17,21-diyl dipropanoate, pregna-l,4-dien-2J-yl propanoate,
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
Beclometasone Dipropionate
Monohydrate
Q. 16p-methyl-3,20-dioxopregna-I,4-diene-17,21-diyl
dipropancare,
H,O
C"H"CIO"H,O 539.1
Action and use

R. 9,11 p-epoxy-17,21-dihydroxy-16Il-methyl-9p-pregna-1,4- Glucocorticoid.
diene-3,20-dione, Preparations
Beclometasone Aqueous Nasal Spray
Beclometasone Inhalation Powder
Beclometasone Inhalation Powder, pre-metered
PhE<I ~ _
DEFINITION
9-Chioro-1I Il-hydroxy-16p-methyl-3,20-dioxopregna-I,4-
dlene-I?,21-diyl dipropanoare monohydrate.
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1-262 Beclometasone Dipropionate Monohydrate 2022
Content Flew rate 1.4 mUrnm..

97.0 per cent to 102.0 per cent (dried substance). Detection Spectrophotometer at 254 nm.
CHARACTERS Injection 20 J.l1 of test solution (a) and reference
Appearance solutions (a), (b) and (c).
White or almost white powder. Identification of impurities Use the chromatogram supplied
Solubility with beclometasone dipropionate for peak identification CRS and
Practically insoluble in water, freely soluble in acetone, the chromatogram obtained with reference solution (c) to
sparingly soluble in ethanol (96 per cent). identify the peaks due to impurities B, C, F and 1..; use the
IDENTIFICATION chromatogram supplied with bedometasone dipropionate for
A. Infrared absorption specrrophotometry (2.2.24). system suitalnliry CRS and the chromatogram obtained with
reference solution (b) to identify the peak due to impurity D.
Comparison bedometasone dipropitJnate monohydrate CRS.
Relativeretention With reference to beclometasone
B. Treat 25 mg by the oxygen-flask method (2.5.11}). Use a
dipropionate (retention time = about 25 min):
mixture of I mL of 1 M sodium hydroxide and 20 mL of impurity B = about 0.6j impurity D = about 1.1j
water R to absorb the combustion products. The solution impurity L = about 1.3; impurity C = about 1.8;
gives reaction (a) of chlorides (2.3.1). impurity F = about 2.2.
C. Loss on drying (see Tests). System suitability Reference solution (b):
TESTS - peak-IO..IlJal/ey ratio: minimum 1.5, where HI' = height
Specific optical rotation (2.2.7) above the baseline of the peak due to impurity D and
+ 108 to + 115 (dried substance). HI) = height above the baselineof the lowest point of the
Dissolve 0.100 g in echanol (96 per cent) R and dilute to curve separating this peak from the peak due to
10.0 mLwith the same solvent. beclometasone dipropionate.
Related substances Limits:
Liquid chromatography (2.2.29). - comaion factor: for the calculation of content, multiply the
So/vem mixture Mobile phase AJ mobilephase B peak area of impurity F by 1.3;
(45:55 VIII). - impun'ty B: not more than 5 times the area of the
examined in 28 mL of mobile phase B and dilute to 50.0 mL
- impun·ties C, F, L: for each impurity, not more than
with mobilephase A.
Test solution (b) Dilute 1.0 mL of test solution (a) to chromatogram obtained with reference solution (a)
50.0 mL with the solvent mixture. (0.15 per cent);
Reference solution (a) Dilute 5.0 mL of test solution (b) to - unspedfied impurities: for each impurity, not more than the
100.0 mL wil:h the solvent mixture. area of the principal peak in the chromatogram obtained
Reference solution (b) Dissolve 5 mg of bedomerasone with reference solution (a) (0.10 per cent);
dipropionate for system suitability CRS (containing impurity D) - totol: not more than 10 times the area of me principal
in 3 mL of mobile phase B and dilute to 5 mL with mobile peak in the chromatogram obtained with reference
phase A. solution (a) (1.0 per cent);
Referen", solution (c) Dissolve 5 mg of bec1omerasone - disregard limit: 0.5 times the area of the principal peak in
dipropionate for peak idenrijication CRS (containing the chromatogram obtained with reference solution (a)
impurities B, C and L) in 3 mL of mobile phase B and dilute (0.05 per cent).
to 5 mL with mobile phase A. Use 1 mL of this solution to Loss on drying (2.2.32)
dissolve me contents of a vial of bedometasone dipropionate 2.8 per cent to 3.8 per cent, determined on 1.000 g by
impuniies F and N CRS. drying in an oven at 105 "C for 3 h.
Reference solution (d) Dissolve 50.0 mg of anhydrous ASSAY
bedometasone dipropionate CRS in 28 mL of mobile phase B Liquid chromatography (2.2.29) as described in the test for
and dilute to 50.0 mL with mobile phase A. Dilute 1.0 mL related substances with the following modification.
of this solution to 50.0 mL with the solvent mixture.
Injection Test 'solution (b) and reference solution (d).
Column:
Calculate the percentage content of C2sHn CI0 7 from the
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: spherical difunctional bonded end-eopped declared content of anhydrous bedometasone dipropionate CRS.
octade<ylsilyl silica gelfor chromatography R (5 um); IMPURITIES
- temperature: 50 "C. Specijied impurities B, C, F, L.
Mobile phase: Ocher detectable impurities (che following substances uould, if
- mobile phase A: 2.72 gIL solution of parassium dihydrogen present at a sufficient level, be detected by oneor other of the tests
plwsphate R adjusted to pH 2.35 with phosphoric acid R; in the monograph. They' are limited by thegeneral aaepUltlU
- mobile phase B: tetrahydro/uran R, acetonitrile R, methanol R criterion for otherlunspecified impurities and/or by che general
(5:23:25 VIVIJI); monograph Substances for pharmaceutical use (2034). It is
Time Mobile phase A Mobile phase B demonstration of complianu. See also 5.10. Control of impurities
(min) (per cent VIJi) (per cent VIV)
in substances for pharmaceutical use) A, D, E, H, I, J, M, N,
0-4 40 60 OJ Q, R, S, U, v.
4 - 12 40 ---> 45 60 --+ 55
12 - 59 45 55
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2022 Beclometasone Dipropionate Monohydrate 1-263
o
°O-Z-CH3
__ '_O~CH3
\ CH 3
0-" '-"'-. L,.,...oJ H
o o \
H B'
A. 9-chloro-ll P,17-dihydroxy-16p-methyl-3,20-dioxopregna-
1,4-dien-21-yl propanoate (beclometasone 21-propionate), F. 6x-bromo-9-chloro-llP-hydroxy-16p-methyl-3,20-
dioxopregna-lA-diene-17,21-diyl dipropanoare,
OH
o
"'_O~CH3
, CH3
~, ~"'" I ,-;--.-/ ""H
B. 21-(acetyloxy)-9-chloro-ll P-hydroxy-16p-methyl-3,20- H. 9-chloro-ll p,21-dihydroxy-16Jl-methyl-3,20-dioxopregna-

dioxopregna-Ld-dien-I'r-yl propancate (beclometasone lA-dien-17-yl propanoate (beclometasone 17-propionate),
zl-acetete 17-propionate),
o
o I. 16p-methyl-3,20-dioxopregna-l,4,9(11)-triene-17,21-<liyl
dlpropanoate,
C. s-chloro-It P-hydroxy-16p-methyl-3,2o--dioxo-I7-
(propanoyloxy)-pregna-l,4-dien-21-yl butanoare
(beclometasone 21 -butyrate 17-propionate),
J. 9,11 Jl-epoxy-16p-methyl-3,20-dioxo-9p-pregna-l,4-diene-
o 17,21-diyl dlprcpanoate,
D. 9-bromo-ll P-hydroxy-16p-methyl-3,20-dioxopregna-l,4-
diene-I?,21-diyl dipropanoate,
o
o O~CH,
o
.... O~CH3
CH, o
H
L. 9-chloro-ll P-hydroxy-16Jl-methyl-3,20-dioxopregn-4-ene-
o 17,21-diyl diprcpanoate,
E. 6x,9-dicbloro-ll p-hydroxy-16p-methyl-3,20-dioxopregna-
l,4-diene-17,21-diyl dipropanoate,
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1-264 Beeswax 2022
o
o °O~CH3
CH3 ~CH3
--0
" CH 3
H
o
S. 9-chloro-I6~-methyl-3,20-dioxopregna-I,4-diene
M. 9-cWoro-11 ~-hydroxy-16~-methyl-3,2o-dioxopregna-4,6
11~,17 ,21-triyl tripropanoate (beclometasone
diene-l ? ,21-diyl dipropanoate,
rripropionate),
o OH
o
°o~CH3 _"O~CH3
···0
~CH3 ", CHJ
, CH3
H
B, e-; /"'-,.' /------' 'H
o
o u. 9,llll-epoxy-21-hydroxy-16~-methyl-3,20-dioxo-9~
pregna-l,4-dien-17 -yl propanoate,
N. 2_hromo_9-cWoro_llll-hydroxy_16~_methyl_3,20_
dioxopregna-l,4-diene-17,21-diyl dipmpanoate,
o
°O~CH3
____ o~CH,
\ CH3
«<: /"'-, /------' H
V. 9,11~_epoxy_17_hydroxy_16~_methyl_3,20_dioxo_9~_
o pregna-I,4-dien-21-yl propanoate.
___________________ Ph,,,,
O. 9,llll-dicWoro-16~-methyl-3,20-dioxopregna-I,4-diene
17,21-diyl dipropanoate,
White Beeswax
Action and use
Excipient.
Ph,,,, _
o
DEFINlTION
Q. 16~-methyl-3,20-dioxopregna-l A-diene-17,21-diyl Wax obtained by bleaching yellow beeswax.
dlpropanoate, CHARACfERS
OH Appearance
White or yellowish-white pieces or plates, translucent when
thin, with a fine-grained, matt and non-crystalline fracture;
when warmed in the hand they become soft and malleable.
It has an odour similar to thatof yellow beeswax, though
fainter and never rancid. It is tasteless and does not stick to
o the teeth.
Soluhility
R. 9,llll-epoxy-17,21-dihydroxy-16~-methyl-9~-pregna-I,4 Practically insoluble in water, partially soluble in hot ethanol
diene-3,2o-dione, (90 per cent VIJI) and completely soluble in fatty and
essential oils.
Relative density
About 0.960.
TESTS
Drop point (2.2.17)
61°C to 66 °C.
Melt the beeswax by heating on a water-bath, pour onto a
glass plate and allow to cool to a semi-solid mass. Fill the
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2022 Beeswax 1-265
metal cup by inserting the wider end into the beeswax and
Yellow Beeswax ***
repeating the procedure until beeswax extrudes from the ** **
narrow opening. Remove the excess with a spatula and insert
*****
the thermometer immediately. Remove the beeswax
displaced. Allow EO stand at room temperature for at least Action and use
12 h before determining the drop point. Excipient.
Acid value PhE" _
17.0 lO 24.0.
DEFINITION
To 2.00 g (m g), in a 250 mL conical flask fitted with a
Wax obtained by melting the walls of the honeycomb made
reflux condenser, add 40 mL of xylene R and a few glass
by the honey-bee, Apis melli/era L.J with hot water and
beads. Heat until the substance is dissolved. Add 20 mL of
removing foreign matter.
etha"ot (96 per "",) Rand 0.5 mL of pheuolphthalei"
solution Rl and titrate the hot solution with 0.5 M alcoholic CHARACTERS
potassium hydroxide until a red colourpersists for at least Appearance
10 s ('" mL). Carry out a blank test ('" mL). Yellow or light brownpieces or plates with a fine-grained,
matt and non-crystalline fracture; when warmed in the hand
Acid I _28_.0_5--'(--'' ,--'-----'",''-) they become soft and malleable.
CL va ue-=
m It has a faint odour, characteristic of honey. It is tasteless and
does not stick to the teeth.
Ester value (2.5.2)
70 to 80. Solubility
Saponification value Practically insoluble in water, partially soluble in hot ethanol
(90 per cent VIV) and completely soluble in fatty and
87 to 104.
essential oils.
To 2.00 g (m g), in a 250 mL conical flask fitted with a
reflux condenser, add 30 mL of a mixture of equal volwnes Relative density
of ethanol (96 per "",) Rand xyIeue R and a few glass beads. About 0.960.
Heat until the substance is dissolved. Add 25.0 mL of 0.5 M TESTS
alcoholic potassium hydroxide and heat under a reflux Drop point (2.2.17)
condenser for 3 h. Titrate the hot solution immediately with 61 "C to 66 "C.
0.5 M hydrochlori< acid, using I mL of pheuolphthalei" Melt the beeswax by heating on a water-bath, pour onto a
solution RJ as indicator (til mL). Reheat the solutionto glass plate and allow to cool to a semi-solid mass. Fill the
boiling several times during the courseof the titration. Carry metalcup by inserting the widerend into the beeswax and
out a blank test (nz mL). repeating the procedure until beeswax extrudes from the
narrow opening. Remove the excess with a spatula and insert
28.05(", - ".)
Saponification value the thermometer immediately. Remove the beeswax
m displaced. Allow to stand at room temperature for at least
Ceresin, paraffi..D.s and certaln other waxes 12 h before determining the drop point.
To 3.0 g, in a 100 mL round-bottomed flask, add 30 mL of Acid value
a 40 f!IL solution of potassium hydroxide R in aldehyde-free 17.0 to 22.0.
akoholR and boil gentlyunder a reflux condenser for 2 h. To 2.00 g (m g), in a 250 mL conical flask fitted with a
Remove the condenser and immediately insert a reflux condenser, add 40 mL of xylene R and a few glass
thermometer. Place the flask in a water-bath at 80 °C and beads. Heat until the substance is dissolved. Add 20 mL of
allow to cool, swirling the solution continuously. etha"ot (96 per ""0 Rand 0.5 mL of pheuolphthalei"
No precipitate is formed until 65 °C, although the solution solution RJ and titrate the hot solution wilh 0.5 M alcoholic
may be slightly opalescent. Beginning at 65 °C, the solution potassium hydroxide until a red colourpersists for at lease
may become cloudyand precipitates may be formed. 10 s ('" mL). Carry out a blank test (n mL).
At 59 °C, the solution is cloudy.
Glycerol and other polyols Acid I
Cl vaue=
28.05(", - "')
Maximum 0.5 per cent mtm, calculated as glycerol. m
To 0.20 g add 10 mL of akohot;,; potassium hydroxide
Ester value (2.5.2)
solution R and heat on a water-bath under a reflux condenser
70 to 80.
for 30 min. Add 50 mL of di/ute sulfioic acidR, cool and
filter. Rinse the flask and the filter with di/ute su/furi< acid R. Saponification value
Combine the filtrate and washings and dilute to 100.0 mL 87 to 102.
with di/ure su/furic acidR. Place 1.0 mL of the solution in a To 2.00 g (m g), in a 250 mL conical flask fitted with a
test-tube, add 0.5 mL of a 10.7 f!IL solution of sodium reflux condenser, add 30 mL of a mixture of equal volumes
periodate R, mix and allow to standfor 5 min. Add 1.0 mL of of etha"ot (96 per ceuV Rand xyIeue R and a few glass beads.
decdorised fuchsin solution R and mix. Any precipitate Heat until the substance is dissolved. Add 25.0 mL of 0.5 M
disappears. Place the tube in a beaker containing water at alcoholic potassium hydroxide and heat under a reflux
40 "C. During cooling observe for 10-15 min. Aoy violet- condenser for 3 h. Titrate the hot solution immediately with
blue colour in the solutionis not more intense than that in a 0.5 M hydrochloric acid, using I mL of phetwlph'haieI"
standard prepared at the same time and in the same manner solution Rl as indicator (nl mL). Reheat the solution to
using 1.0 mL of a 10 mf!IL solution of glycerol R in di/ute boiling several times during the course of me titration. Carry
sulfuric acidR. out a blank test ('" mL).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
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1-266 Benazepril Hydrochloride 2022
==='----=-
icauon value = 28.05(n, - nIl
Sapomificati
m
IDENflFICATION
Carry out either tests A, BJ D or tests B, CJ D.
Ceresin, paraffins and certain other waxes A. Specific optical rotation (2.2.7): -141 to -136 (dried
To 3.0 g, in a 100 mL round-bottomed flask, add 30 mL of substance).
a 40 gIL solution of potassium hydroxide R in aldehyde-free Dissolve 1.000 g in anhydrous ethanol R and dilute to
akolw/R and boil gently undera reflux condenser for 2 h. 50.0 mL with the same solvent"
Remove the condenser and immediately inserta B. Infrared absorption spectrophotometry (2.1.24).
thermometer. Place the flask in a water-bath at 80°C and
Comporison benazepril hydrochloride CRS.
allow to cool, swirling the solutioncontinuously.
No precipitate is formed until 65 °C, although the solution If me spectra obtained in the solid state show differences,
may be slightly opalescent. Beginning at 65 QC, the solution dissolve the substance to be examined and the reference
may become cloudyand precipitates may be formed. substance separately in methanol R, evaporate to dryness and
At 59°C, the solution is cloudy. record new spectra using me residues.
C. Enantiomeric purity (see Tests).
Glycerol and other polyols
Maximum 0.5 per cent mtm, calculated as glycerol. D. It gives reaction (a) of chlorides (2.3.1).
To 0.20 g add 10 mL of alcoholic potassium hydroxide TESTS
so/mum R and heat on a water-bath under a reflux condenser Related substances
for 30 min. Add 50 mL of dilu.. sulfuric acid R, cool and Liquid chromatography (2.2.29).
filter. Rinse the flask and the filter with d.7u te sulfuric acidR. Testsolution (a) Dissolve 50.0 mg of the substance to be
Combine the filtrate and washings and dilute to 100.0 mL examinedin the mobile phase and dilute to 50.0 mL with
with dilu.. su/juri< acidR. Place 1.0 mL of the solution in a the mobile phase.
test-tube, add 0.5 mL of a 10.7 gIL solution of sodium
Tesr solurion (b) Dilute 10.0 mL of test solution (a) to
periodate RJ mix and allow to stand for 5 min. Add 1.0 mL of 100.0 mL with the mobile phase.
deco/orised fuchsin solution R and mix. Any precipitate
disappears. Place the tube in a beaker containing water at Reference solution (a) Dissolve 50.0 mg of benazepril
40°C. During cooling observe for 10-15 min. Any violet- hydrochloride CRS in the mobile phase and dilute to 50.0 mL
blue colour in the solution is not more intense than thatin a with the mobile phase. Dilute 10.0 mL of this solution to
standard prepared at the same time and in the same manner J 00.0 mL with the mobile phase.
using 1.0 mL of a 10 mgIL solution of glycerol R in diluze Referena solution (b) Dissolvethe contents of a vial of
su/juric acid R. benazepril for syste m suitability CRS (containing impurities B,
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEII C, D, E, F and G) in 1.0 mL of test solution (a).
Reference solurion (c) Dilute 1.0 mL of reference solution (a)
Column:
Benazepril Hydrochloride - size: I ;;;; 0.30 m, 0 ;;;; 3.9 mm;
- stationary phase: end-capped octode<YfsiIYI silica gelfor
(Ph. Eur. monograph 2388) chromatography R (10 urn},
Mobile phase Add 0.2 mL of glacial aceric acidR to
1000 mL of a mixture of 360 volumes of water Rand
640 volumes of methanol R2; add 0.81 g of
• He! tetrabutylammonium bromide R and stir to dissolve"
Flow ra" 1.0 mUmin.
lnjecn"on 25 J.IL of test solution (a) and reference
461.0 86541-74-4 solutions (b) and (c).
Action and use Run time 3 times the retention time of benazepril,
Angiotensin converting enzyme inhibitor. Relative retention With reference to benazepril (retention
time e about 6 min): impurity E = about 0.3;
impurity F =about 0.4; impurity C =about 0.5;
PhEII _
DEFINITION impurity B = about 1.8; impurity D = about 2.0;

[(3S) -3- [[(I S)-I-(Ethoxycarbonyl)-j-phenylpropyl] amino]-2- =
impurity G about 2.5.
oxo-2J3J4,5-tetrahydro-IH-l-benzazepin-I-y I] aceticacid Identification of ;mpun"ties Use the chromatogram supplied
hydrochloride. with benazepril for SYSI4m suitabih'ty CRS and the
Content chromatogram obtainedwith reference solution (b) to
97.5 per cent to 102.0 per cent (dried substance). identify the peaks due to impurities B, C, D, E, F and G.
System SUiUlbility Reference solution (b):
CHARACTERS - resolution: minimum 2.5 between the peaks due to
Appearance benazepril and impurity B and minimum 1.5 between the
White or almost white, crystalline powder, hygroscopic. peaks due to impurities E and F.
Solubility Limits:
Slightly soluble in water, freely soluble in anhydrous ethanol, - correction !lUlOTS: for the calculation of content, multiply
veryslightly soluble in ethyl acetate, practically insoluble in the peak areas of the following impurities by the
cyclohexane.
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2022 Benazepril Hydrochloride 1-267
corresponding correction factor: impurity E = 0.5; Loss on drying (2.2.32)

=
impurity F 0.7; Maximum 1.5 per cent) determined on 1.000 g by drying in
- impun'ty B: not more than 2.5 times the area of the vacuo at 105 "C for 3 h.
principal peak in the chromatogram obtained with Sulfated ash (2.4.14)
reference solution (c) (0.5 per cent); Maximum 0.1 per cent) determined on 1.0 g.
- impurity C: not more than 1.5 times the area of the
principal peak in the chromatogram obtained with ASSAY
reference solution (c) (0.3 per cent); Liquid chromatography (2.2.29) as described in the test for
- impurities D, E, F, G: for each impurity, not more than related substances with the following modification.
the area of the principal peak in the chromatogram Injection Test solution (b) and reference solution (a).
obtained with reference solution (c) (0.2 per cent); Calculate the percentage content OfC24H29ClN20j from the
- unspedfied impmities: for each impurity, not more than declared content of benazepril hydrochloride CRS.
STORAGE
(0.10 per cent); Protected from light, in an airtight container.
- total: not more than 10 times the area of the principal IMPURlTIES
peak in the chromatogram obtained with reference Specified impuruies A, B, G, D, E, F, G.
- disregard Urnit: 0.25 times the area of the principal peak in
(0.05 per cent).
Enantiomeric purity
Buffer solutien pH 6.0 Dissolve 3.58 g of diwdium hydrogen
phosphate dodecahydrate R and 9.66 g of potassium dihydrogen A. [(3R)-3-[[(IR)-I-(ethoxycarbonyl)-3-phenylpropyl]amino)-
phosphate R in waterR and dilute ro 1000.0 mL with the 2-oxo-2)3)4,5-tetrahydro- t H-l-benzazepin-l-yl] acetic
same solvent. acid,
Ten solution Dissolve 50.0 mg of the substance to be
the mobile phase.
Reference solution (a) Dissolve 5.0 mg of benazeptil
impurity A CRS in the mobile phase and dilute ro 50.0 mL
Reference sol. .ion (b)Dilute 1.0 mL of reference solution (a)
to 100.0 mL with the mobile phase. B. [(3RS)-3-[[(ISR)-I-( ethoxycarbonyl)-3-
phenylpropyl] amino]-2-oxo-2,3,4,5-tettahydro-l H-I-
benzazepin- J-yl]acetic acid)
ro 10.0 mL with the mobile phase. Dilute 1.0 mL of this
solution to 10.0 mL with the test solution.
Column:
- size: 1= 0.10 m, 0 = 4.0 mm;
- stationary phase: spherical silica gelAGP for chiral
chromatography R (5 urn);
Mobile phase methanol R2, buffer solution pH 6.0
(20:80 VII'). c. (2S)-2-[[(3S)-1-(carboxymethyl)-2-oxo-2,3,4, 5-tetrahydro-
IH-I-benzazepin-3-yl]amino)-4-phenylbutanoic acid,
Deteaion Spectrophotometerat 240 nm.
Injection 50 J.1L of the test solution and reference
Run time 3.5 times the retention time of benazepril.
Relative retention With reference to benazepriJ (retention
time = about 6 min): impurity A = about 1.9.
System suitability Reference solution (c): D. [(3S)-3-[[(IS)-3-cyclohexyl-I-(ethoxycarbonyl)
- peak-ro-vaUey ratio: minimum 2.5, where Hp = height propyl]amino)-2-oxo-2,3,4,5-tettahydro-IH-I-benzazepin-
above the baseline of the peak due to impurity A and l-yl]acetic acid,
HI} = height above the baseline of the lowest point of the
curve separating this peak from the peak due to H0
benazepril. H~tsC02H
Limit:
- impurity A: not more than the area of the corresponding
E. [(3S)-3-amino-2-oxo-2,3,4,5-tetrahydro-lH-l-benzazepin-
l-yl]acetic acid)
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1-268 Bendroflumethiazide 2022
Reference solution (a) Dissolve 2 mg of bendrojlumethiazide

impurity A CRS and 2.5 mg of altizide CRS in the solvent
mixture and dilute to 10 mL with the solvent mixture.
&lix 1 rnL of this solution with 1 mL of the test solution and
dilute to 100 mL with the solvent mixture.
F. I,I-<limethylethyl [(3S)-3-amino-2-oxo-2,3, 4,5-tetrahydro- 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
I H-I-benzazepin-l-yl] acetate, solution to 10.0 mL with the solventmixture.
Column:
- size: 1== 0.15 m, 0 :::: 3.0 mm;
- stationary phase: end-capped oaadecylsiJyl silica gelfor
chromatography R (5 um);
Mobile phase Mix IS volumes of telrahydrofuran R,
25 volumes of methanol Rand 60 volumes of a 2.0 !¥L
G. ethyl (2S)-2-[[(3S)-I-(2-ethoxy-2-<>xoethyl)-2-oxo-2,3,4,5- solutionof cimc acid monohydrate R.
tetrahydro-I H-l-benzazepin-3-yl]amino]-4-
phenylbutanoare.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'IIE"
Injection 20 ~L.
Run time Twice the retention time of bendroflumethiazide.
Relative retention With reference to bendroflumethiazide
Bendroflumethiazide (retention time :::: about 8 min): impurity A::::: about 0.2;
altizide =about 0.5.
(Ph. Eur. monograph 0370) System suitability Reference solution (a):
- resolution: minimum 10 between the peaksdue to ahizide
o\\ 0 0 0
S
II \\.(1
S . and bendroflumethiazide.
H,N' YY 'NH))'" andenanUomer Limits:
FC~N-1"
• H H
# - impurity A: not more than the area of the principal peak
in the chromatogram obtainedwith reference solution (b)
(0.1 per cent);
421.4 73-48-3 - unspecified impurities: for each impurity) not more than the
Action and use with reference solution (b) (0.10 per cent);
Thiazide diuretic. - total: not more than twice the area of the principal peak in
Preparations the chromatogram obtainedwith reference
Bendrollumethiazide Tablets solution (b) (0.2 per cent);
Bendroflumethiazide OralSuspension - disregard limit: 0.5 times the area of the principal peakin
I'IIE" _
(0.05 per cent).
DEFINITION Loss on drying (2.2.32)
(3RS)-3-Benzyl-6-(trif1uoromethyl)-3,4-dihydro-2H-I,2,4- Maximum 0.5 per cent, determined on 1.000 g by drying in
benzothiadiazine-7-sulfonamide 1,1-dioxide. an oven at 105°C.
Content Sulfated ash (2.4.14)
98.0 per cent to 102.0 per cent (dried substance). Maximum 0.1 per cent, determined on 1.0 g.
CHARACTERS ASSAY
Appearance Dissolve 0.150 g in 50 mL of dimethyl sulfoxide R. Titrate to
White or ahnost white, crystalline powder. the 2nd point of inflexion with 0.1 M tetrabutylammonium
Solublllry hydroxide in 2-propanol, determining the end-point
Practically insoluble in water, freely soluble in acetone, potentiometrically (2.2.20). Carry out a blank titration.
soluble in ethanol (96 per cent). I mL of 0.1 M tetrabutylammonium hydroxide in 2-propanol is
equivalent to 21.07 mg OfClSHI4F3N304S2.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24). IMPURITffiS
Comporison bendrcfiumednazide CRS. Specified impurities A.
TESTS
Related substances
Solvent mixture Mix 40 volumes of methanol Rand
60 volumes of a 2.0 gIL solution of dm', acid monohydrate R. A. 4-amino-6-(trifluoromethyl) benzene-I)3-disulfonamide.
Test solution Dissolve 10.0 mg of the substance to be _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
examined in the solventmixture and dilute to 50.0 mL with
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2022 Benperidol 1-269
*** c. Dissolve about 10 mg in 5 mL of anhydrous ethanol R.

Benperidol
*** *** Add 0.5 mL of dinitrobenzene solution Rand 0.5 mL of 2 M
(Ph. Eur. monograph 1172) *** akoholic potassium hydroxide R. A violet colour is produced
which becomes brownish-red after 20 min.
D. Mix about 5 mg with 45 mg of heavy magnesium oxide R
O)-NH
and ignite in a crucible until an almost while residueis
o (yN~ obtained (usually less than 5 min). Allow to cool, add 1 mL
~N-./ U of water R, 0.05 mL of phenolphthalein solution Rl and about

I mL of dilutehydrochloric acid R to render the solution
F V colourless. Filter. To a freshly prepared mixture of 0.1 mL of
a/izan'n S solution Rand 0.1 mL of zirconyl nitrate solution R)
add 1.0 mL of the filtrate. Mix, allow to standfor 5 min and
381.4 2062-84-2 compare the colour of the solution with thatof a blank
prepared in the same manner. The test solution is yellow and
Action and use
the blank is red.
TESTS
PhE" _
Related substances
DEFINITION Liquid chromatography (2.2.29). Prepare the solutions
1-[ 1-[4-(4-Fluorophenyl)-4-oxobutyl) piperidin-4-yl)-1,3- immet/iarely before use.
dihydro-2H-benzimidazol-2-one. Test solution Dissolve 0.10 g of the substance to be
Content examined in dimethy!fonnamide R and dilute to 10.0 mL with
99.0 per cent to 101.0 per cent (dried substance). the same solvent.
Reference solution (a) Dissolve 2.5 mg of benperidDl CRS and
CHARACTERS
2.5 mg of droperidol CRS in dimethy!formamide R and dilute to
Appearance 100.0 mL with the same solvent.
Solubility 100.0 mL with dimethy!formamide R. Dilute 5.0 mL of this
Practically insoluble in water, freely soluble in solution to 20.0 mL with dimethy!formamide R.
dimethylformamide, soluble in methylene chloride, slightly
Column:
- size: 1= 0.101,0 = 4.6 mm;
It shows polymorphism (5.9). - stationary phase: base-deaaitxued octade<y/siIYI silica gelfur
IDENTIFICATION chromatography R (3 urn).
First identification: A. Mobile phase:
Second identification: B, C, D. - mobile phaseA: 10 gIL solution of letTaburylammonium
A. Infrared absorption spectrophotometry (2.2.24). hydrogen sulfate R;
- mobile phase B: acetonitrile Rj
Comporison benpeidd CRS.
If the spectra obtained in the solid state show differences, Time Mobile phase A MobUe phase B
dissolve the substance to he examined and the reference (min) (per cent V/Jo? (per cent V/l?
substance separately in the minimum volume of mezhyl 0-15 100 -+ 60 o --J 40
isobutyl ketone R, evaporate to dryness and record new spectra 15 - 20 60 40
using the residues. 20 - 25 100 0
Tesl so/urian Dissolve 30 mg of the substance lO be Flow rate 1.5 mUmin.
examined in the mobile phase and dilute to 10 mL with the Detection Spectrophotometer at 275 run.
mobile phase. Injection 10 ~.
Reference sdution (a) Dissolve 30 mg of benjMridDl CRS in Relat1ve mention With reference to benperidol (retention
the mobile phase and dilute to 10 mL with the mobilephase.
time = about 6.5 min): impurity A = about 0.2;
Reference solution (b) Dissolve 30 mg of benpetidol CRS and impurity B = about 0.9; droperidol = about 1.1;
30 mg of dropeniWl CRS in the mobile phase and dilute to impurity D = about 1.2; impurity E = about 1.3;
10 mL with the mobile phase. =
Plate TLC silica gelFm plate R. System suitability Reference solution (a):
Mobile phase acetone R, methanolR (10:90 VIV). - resolution: minimum 2.0 between the peaks due to
Application 10 ~L. benperidol and droperidol.
Development Over 3/4 of the plate. Limits:
Drying In air.
- impurities A, B, C, D, E: for each impurity, not more than
the area of the principal peakin the chromatogram
Detection Examine in ultraviolet light at 254 nm. obtainedwith reference solution (b) (0.25 per cent);
System suitability Reference solution (b): - unspecified impuniies: for each impurity, not more than
- the chromatogram shows 2 clearly separated spots. 0.4 times the area of the principal peak in the
Results The principal spot in the chromatogram obtained chromatogram obtained with reference solution (b)
with the test solution is similar in position and size to the (0.10 per cent);
solution (a).
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1-270 Benserazide Hydrochloride 2022
(0.5 per cent);
(0.05 per cent).
E. trans-I- [1-[4-(4-lIuorophenyl)-4-oxobutyl]piperidin-4-yl
I-oxide]-1, 3-dihydro-2H-benzimidazol-z-one.
an oven at 105°C.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""E<I
Maximum 0.1 percent, determined on 1.0 g in a platinum
crucible.
ASSAY Benserazide Hydrochloride
Dissolve 0.300 g in 50 mL of a mixture of 1 volume of
anhydrous ace"cacid Rand 7 volumes of methyl ethylkaone R. (ph. Hur. monograph 1173)
Titrate with 0.1 M perch/oric acid, using 0.2 mL of
naphtho/benzein solution R as indicator. H H .N~
q::
I mL of 0.1 M perchloric acid is equivalent to 38.14 mg N ' N ~OH
H andenantlomer • Hel
of C22H24FN302' HO~ OH 0
STORAGE OH
IMPURITIES 293.7 14919-77-8
Sp«ified im[mrilies A, B, C, D, H.
Action and use
Dopa decarboxylase inhibitor.
Preparations
Co-benetdopa Capsules
Co-beneldopa Dispersible Tablets
Co-beneldopa Prolonged-release Capsules
A. l-cpiperidin-4-yl)-1,3-dihydro-2H-benzimidazol-2-one, PIIE<I _
DEFINITION
(2RS)-2-Amino-3-hydroxy-N'-[(2,3,4-trihydroxyphenyl)
methyl]propanehydrazide hydrochloride.
Content
CHARACTERS
B. 1-[1- [4-(2-lIuorophenyl)-4-oxobutyl)piperidin-4-yl]-1,3- Appea:rance
dihydro-2H-benzimidazol-2-one, White or yellowish-white or orange-white, crystalline powder.
Solubility
Freelysoluble in water, very slightly soluble in anhydrous
ethanol, practically insoluble in acetone.
IDENTIFICATION
Comparison benserazide hydrochloride CRS.
If the spectra obtained show differences, dissolve the
substance to be examined and the reference substance
separately in hot methanol R, evaporate to dryness and record
new spectra using the residues.
C. 1-[I-[4-oxo-4-[4-[4-(2-oxo-2,3-dihydro-IH-benzimidazol-
I-yl) piperidin-I-yl] phenyl] butyl] piperidin-4-yl)-1,3- B. Dissolve 16 mg in 2 mL of methanol R. The solution gives
dihydro-2H-benzimldazoJ-2-one, reaction (a) of chlorides (2.3.1).
TESTS
Solution S
Dissolve 1.0 g in carbon dioxide-fru walel'R and dilute to
than reference solution BY. (2.2.2, Method II).
D. cis-I- [1-[4-(4-lIuorophenyl)-4-oxobutyl] piperidin-4-yl pH (2.2.3)
l-oxide]-1,3-dihydro-2H-benzimidazol-2-one, 4.0 to 5.0 for solution S.
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2022 Benserazide Hydrochloride 1-271
Related substances - ;mpun',y C: not more than the area of the corresponding
Liquid chromatography (2.2.29). AU solutions must be injected peak or pair of peaks in the chromatogram obtained with
immediately or stored at 4 °G. reference solution (b) (0.5 per cent);
Test solution Dissolve 0.100 g of the substance to be - unspecified impurities: for each impurity) not more than the
examined in methanol R and dilute to 50.0 mL with the same area of the principal peak in the chromatogram obtained
solvent. with reference solution (a) (0.10 per cent);
- sum of impurities other than A: not more than 10 times the
100.0 mL with methanol R. Dilute 1.0 mL of this solution to
- disregard limit, 0.5 times the area of the principal peakin
Reference solution (b) Dissolve 5.0 mg of benserazide the chromatogram obtainedwith reference solution (a)
impurity A CRS and 5.0 mg of benserazide impurity C CRS in (0.05 per cent).
methanol R and dilute to 50.0 mL with the same solvent.
Dilute 1.0 mL of the solution to 10.0 mL with methanol R. Water (2.5.12)
Maximum 1.0 per cent, determined on 0.500 g. Use as the
Reference solution (c) Dissolve 5 mg of benserazide for peak
solvent a solution containing 30 mL of formamide R, 30 mL
identification A CRS (containing impurity B) in 5 mL of
of methanol Rand 7.0 g of salicylic acidR (the salicylic acidR
reference solution (b). must be added to the solvent mixture in the titration vessel).
Column:
- size: 1= 0.25 m, 0 = 4 mm;
- stationary phase: octylsilyl silica gelfor chromatography R
(5 pm): ASSAY
- temperature: 30 "C. In order to avoid overheating during the titration, mix thoroughly
Mobile pkase: throughout and swp the titration immediately afterthe end-point
- mobile phase A: dissolve 2.2 g of sodium heptonesulfonate has been reached.
monohydrate Rand 6.8 g of potassium dihydrogen Dissolve 0.250 g in 5 mL of anhydrous formic acidR.
phosphate R in 900 mL of waterfor chromatography R, add Add 70 mL of anhydrous acetic acid R. Titrate immediately
50 mL of methanol R2 and adjust to pH 3.5 with with 0.1 M perch/oric acid, determining the end-point
phosphoric acidR; potentiometrically (2.2.21J).
- mobile phase B: dissolve 2.2 g of sodium heptonesulfonare 1 mL of 0.1 M perchloric acid is equivalent to 29.37 mg of
monohydrare Rand 6.8 g of potassium dihydrogen C,oH16C1N3 0,.
phosphore R in 500 ",L of waterfor chromatography R,
adjust to pH 3.5 with phosphoric acid R and add 500 mL STORAGE
of methanol R2j Protected from light.
IMPURITIES
Tim. Moblle phase A Moblle phBlle B
0- 15 100 -i 0 0--->100
15 - 25 o '00
andonanliomer
Fb>w rare 1.3 mUmin.

A. (2RS)-2-amino- 3-hydroxypropanehydsazide,
Injection 5 ~L.
Identification of imjnnltres Use the chromatogram obtained OH
impurities A and C; use the chromatogram supplied with
benserazide for peak identification A CRS and the
the peak due to impurity B; doublingof the peak due to
::'L:J~OH
0 N~ H
end eoeoucmer
HO~H
impurity C, related to separation of the (EZ)-isomers, may be
observed.
Relativeretention With reference to henserazide (retention 0
OH
impurity C = about 1.2; impurity B = about 1.5.
B. (2RS)-2-amino-3-hydroxy-N',N'-bis[(2,3,4-
trihydroxyphenyl)methyl]propanehydrazide,
benserazide and impurity C; use the 1Sl peakof
impurity C if 2 peaks occur.
Limits:
- correction factor: for the calculation of content, multiply the its(Z}-Isomer and
IheirenanUomers
peak area of impurity B by 0.7;
- impun'ty A: not more man the area of the corresponding
C. (2RS)-2-amino- 3-hydroxy-N' -[(EZ)-(2,3,4-
- impuniy B: not more than 5 times the area of the
trihydroxyphenyl)methylidene]propanehydrazide.
reference solution (a) (0.5 per cent); ______________ ~ ""E"
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1-272 Bentonite 2022
mandatory part is included in the FunctilJnality-related

Bentonite charaaeiuia section. Control of the characteristics can contribute
to the qualityof a medicinal product by improving the consistency
of the manufactun'ng process and the perJomlance of the medicinal
PhE<r _
product during use. Where control methods are dud, they are
DEFINITION recognised as being suitable for tht purpose, but othermethods can
Natural clay containing a high proportion of montmorillonite, also be used. Wherever results for a particular characteristic are
a native hydrated aluminium silicate in which some reported, the centrol method must be indicated.
aluminium and silicon atoms may be replaced by other atoms Thefollowing characteristks may be relevant lor bentonite usedas
such as magnesium and iron. wcesity-increasing agent or suspending agent.
CHARACTERS Sedlmentadon volume
Appearance To 6.0 g add 200 mL of waterR and mix for 20 min using a
Very fine, homogeneous, greyish-white powder with a more high-speed mixer capable of operatiog at 10000 rlmin.
or less yellowish or pinkish tint. Transfer 100 mL of this suspension to a graduated cylinder.
Allow to stand for 24 h. The volume of the clear supernatant
Solubility
is not greater than 2 mL.
Practically insoluble in water and in aqueous solutions.
Swelling power with water
It swells with a little water forming a malleable mass.
See Identification B.
IDENTIFICATION _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
A. To 0.5 g in a metal crucible add 1 g of potassium nitrate R
and 3 g of sodium carbonate R and heat until the mixture
melts. Allow to cool. To this residue add 20 mL of boiling
water R, mix and filter. Wash the insoluble residue with
50 mL of waterR. To this residue add I mL of hydrochlori<
Benzaldehyde
acidR and 5 mL of waterR. Filter. To the filtrate add I mL
of strong sodium hydroxide solution R and filter. To this filtrate CHO
add 3 mL of ammoniumchloride solution R. A gelatinous white
B. Add 2.0 g in 20 portions to 100 mL of a 10 gIL solution
of sodium lauri/sulfate R in a 100 mL graduated cylinder
6
about 30 mm in diameter. .Allow 2 min between additions 106.1 100-52-7
for each portion to settle. Allow to stand for 2 h.
The apparent volume of the sediment is not less than 22 mL. Action and use
C. 0.25 g gives the reaction of silicates (2.3.1). Flavour.
TESTS DEFINITION
A1ka1lnlty Benzaldehyde contains not less than 98.0% w/w and not
To 2 g add 100 mL of carbou dioxide-free waterR and shake more than 100.5% w/w ofC7H6 0 .
for 5 min. To 5 mL of this suspension add 0.1 mL of
rhymolphrhalein solution R. The liquid becomes bluish. CHARACTERISTICS
Add 0.1 mL of 0.1 M hydnochl<Jric acid. The liquid is A clear, colourless liquid.
decolourised within 5 min. Slightly soluble in water, miscible with ethanol (96%) and
Coarse particles with ether.
Maximum 0.5 per cent. TESTS
To 20 g add 1000 mL of water R and mix for 15 min using a Refractive index
high-speed mixer capable of operating at not less than 1.544 to 1.546, Appendix V E.
5000 c/min. Transfer the suspension to a wet sieve (75), Weight per mL
rared after drying at 100-105 "C. Wash with 3 quantities, 1.043 to 1.049 g, Appendix V G.
each of 500 ml., of water R, ensuring that any agglomerates
Free acid
have been dispersed. Dry the sieve at 100-105 °C and weigh.
Not more than 1.0% w/v, calculated as benzoic acid,
The particles on the sieve weigh a maximum of 0.1 g.
C,H602, when determined by the following method.
Loss on drying (2.2.32) To 10 mL add 20 mL of ethanol (96%) previously
Maximum 15 per cent, derenuined on 1.000 g by drying in neutralised to phenolphthalein solution R1 and titrate with 0.1M
an oven at 105 "C. sodium hydroxide VS using phenolph.halein solutiou Rl as
Microbial contamination indicator. Each mL of O.lM sodium hydroxide VS is equivalent
TAMC: acceptance criterion 10 3 CFUlg (2.6.12). to 12.21 mg of C,H,02'
FUNCTIONALITY-RELATED CHARACTERISTICS Chlorinated compounds
This section provides information on charaaeristics that are NOl more than 0.05% w/v, calculated as CI, when
recognised as being relevant control parameters for oneor more determined by the following method. To 5 mL add 50 mL of
functions of.m substance uihen used as an excipien: (see chapter isoamyl alcohol and 3 g of sodium and boil under a reflux
5.15). Some of the characteristics described in the FunctionahbJ- condenser for 1 hour. Cool, add 50 mL of water and 15 mL
related characteristics section may alsobepresent in the mandauny of nim'c acid, cool, add 5 mL of 0.1M silver nitrate VS, shake
pan of the monograph since they also represent mandatory quah"cy and titrate the excess silver nitrate with O.1M ammonium
cntetia: In such cases, a cross-reference to the tests described in the thiocyanate VS using ammonium iron(llI) sulfate solution R2 as
indicator. Repeat the procedure without the substance being
www.webofpharma.com
2022 Benzalkonium Chloride 1-273
examined. The difference between the titrations represents principal peaks in the chromatogram obtained with the
the amount of silvernitrate required. Each mL of O.lMsilver reference solution.
nitrate VS is equivalent to 3.545 mg of CI. C. To 2 mL of solution S (see Tests) add 0.1 mL of glaciol
ASSAY acetic mid Rand, dropwise, I mL of sodium tetraphenylbm'are
Carry out the method for determination of aldehydes, sohuion R. A white precipitate is formed. Filter. Dissolve the
Appendix X K, using 0.5 g. Each mL of 0.5M potassium precipitate in a mixtureof 1 mL of acetone R and 5 mL of
hydroxide in ethanol (60%) VS is equivalent to 53.06 mg of ethanol (96 per cent) R, heating to not more than 70 'C.
C,H.O. Add water R dropwise to the wann solution until a slight
opalescence forms. Heat gently until the solution is clear and
STORAGE aUow to cool. White crystals separate. Filter, wash with
Benzaldehyde should he kept in a well-filled container, 3 quantities, each of lO mL, of waterR and dry in vacuo
protected from light and stored at a temperature not (2.2.32) at a temperature not exceeding 50 °C. The crystals
exceeding 15°. melt (2.2.14) at 127 -c to 133 'C.
D. To 5 mL of dilutesodium hydroxide solution R add 0.1 mL
of bromophenol blue solution RI and 5 mL of melhylene
chloride R and shake. The methylene chloride layeris
Benzalkonium Chloride ***
** ** colourless. Add 0.1 mL of solution S and shake.
The methylene chloride layerbecomes blue.
(Ph. Eur. monograph 0372) *****
R. To 2 mL of solution S add 1 mL of dilute nimc acid R.
A white precipitate is formed which dissolves on the addition
CI"
of 5 mL of ethanol (96 per cen!! R. The solution gives
reaction (a) of chlorides (2.3./).
TESTS
8001-54-5 Solution S
Dissolve l.O g in carbon dioxide-free water R and dilute to
Action and use 100 mL with the same solvent.
Antiseptic. Appearance of solution
PIlE" _
than reference solution Y6 (2.2.2, MaJwd II).
DEFINITION Acidity or alkalinity
Mixture of alkylbenzyldin7ethylammonium chlorides, the To 50 mL of solution S add 0.1 mL of bromocresol purple
alkyl groups mainlyhavingchainlengths of e 12, C14 and solution R. Not more than 0.1 rnL of O.J.M hydrochlmi< acid
C 16 · or 0.1 M sodium hydroxide is required to change the colour of
Content the indicator.
95.0 per cent to 104.0 per cent of Average relative molecular mass and ratio of alkyl
alkylbenzyldimethylammonium chlorides (anhydrous components
substance) calculated using the average relative molecular Liquid chromatography (2.2.29).
mass (see Tests),
CHARACTERS examined in water R and dilute to 100.0 mL with the same
Appearance solvent.
White or yellowish-white powderor gelatinous, yellowish- Reference solution Dissolve the contents of a vial of
white fragments, hygroscopic. On heating it forms a clear benzalkoniumchloride for system suitability CRS in 5 mL of
molten mass. water R.
Solubility Column:
Very soluble in waterand In ethanol (96 per cent). - size: I ;;;; 0.25 rn, 0 ;;;; 4.6 mm;
An aqueous solution froths copiouslywhen shaken. - stationary phase: end-capped cyanosJ1yl silica gelfor
IDENTIFICATION chromalcgraphy R (5 urn).
First identification: B, E. Mobile phase Mix 45 volumes of acetonitrile Rand
55 volumes of a 13.6 gIL solution of sodium aarate R
Second identification: A, C, D, E.
previously adjusted to pH 5.0 with glacial ocetic acid R.
(2.2.25). FIuw rate 2.0 mIJmin.
Test solution Dissolve 80 mg in water R and dilute to Detection Spectrophotometer at 254 run.
100.0 mL with the same solvent. Injection 10 ~.
Specual range 220-350 om. Identification of homologues Use the chromatogram supplied
Absorption maxima At 257 om, 263 om and 269 om.
with benzalkonium chloride for system suitability CRS and the
chromatogram obtained with the reference solution to
Shoulder At about 250 om. identify the peaks due to C I 2.1 C I 4 and C 16 .
B. Examine the chromatograms obtained in the test for Relative retention With reference to C I2 homologue
average relative molecular mass and ratioof alkyl (retention time e about 6 min): C 14 homologue > about 1.3;
components. C 16 homologue e about 1.7.
Results The principal peaks in the chromatogram obtained
with the test solution are similar in retention time to the
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1-274 Benzalkonium Chloride 2022
System suitability Reference solution: Flow rate 1.0 mUmin.

- resolution: minimum 1.5 between the peaks due to the C l 2 Detection Spectrophotometer at 210 run for impurities A
and C l 40 homologues. and C) and at 257 nm for impurity B.
Calculate the average relative molecular mass of the sample Injection 20 IJL.
by summing the products for each homologue, using the Relative retention With reference to impurity A (retention
following expression:
=
time about 10 min): impurity B = about 1.3,
impurity C =about 2.4.
w(~) System suitability At 210 nm:
A area of the peak due to the given homologue in the the chromatogram obtained with reference solution (c)j
chromatogram obtained with the test solution;
- symmetryfaaor: minimum 0.6 for the peak due to
B sum of the areas of the peab due [0 all homologues in the
chromatogramobtained with the lest sotuOODj impurity A in the chromatogram obtained with reference
W relative molecular mass for the given homologue: 340, 368 and solution (a).
396 for the C I2J C 14 and C l 6 homotogues, respectively. Limits:
- correction factor: for the calculation of content, multiply the
Calculate the percentage of each homologue, using the peak area of impurity C by 1.3;
following expression: ~ impuniy A: not more than the area of the corresponding
100(~) solution (a) (0.5 per cent);
- impun·ty B: not more than the area of the corresponding
C product of the relative molecular mass of the given homologue peak in the chromatogram obtained with reference
and the area of the cerrespending peak in the chromatogram solution (b) (0.15 per cent);
obtained with the test solution; - impun·/)' G: not more than 0.1 times the area of the
D sum of the C values for all homologues quantified.
Limits: reference solution (a) (0.05 per cent).
- C12 homologue: minimum 40 per cent; Amines and amine salts
- C H homologue: minimum 20 per cent, Dissolve 5.0 g with heating in 20 mL of a mixture of
- sum of ell and eN homologues: minimum 70 per cent. 3 volumes of 1 M hydrochlori< acid and 97 volumes of
Impurities A, Band C methanolR and add 100 mL of 2-propanol R. Pass a stream
of nitrogen R slowly through the solution. Titrate with up to
Liquid chromatography (2.2.'29). Prepare the solutions
immediouly before use. 12.0 mL of 0.1 M tesrobutylammonium hydroxide and record
the potentiometric titration curve (2.2.20). If the curve shows
Test solution Dissolve 0.500 g of the substance to be 2 points of inflexion) the volume of titrant added between the
examined in methanolR and dilute to 10.0 mL with the same
2 points is not greater than 5.0 mL. If the curve shows no
solvent.
point of inflexion, the substance to be examined does not
Reference solulion (a) Dissolve 25.0 mg of benzyl akohol CRS comply with the test. If the curve shows 1 point of inflexion,
(impurity A) in methanolR and dilute to 100.0 mL with the repeat the test but add 3.0 mL of a 25.0 gIL solution of
same solvent dimethyldecy/amine R in 2-propanol R before the titration.
Reference solution (b) Dissolve 75.0 mg of benzaldehyde CRS If the titration curve after addition of 12.0 mL of the titrant
(impurity B) in methanol R and dilute to 100.0 mL with the shows only 1 point of inflexion) the substance to be
same solvent. Dilute 1.0 mL of this solution to 10.0 mL with examined does not comply with the test.
methanol R. Water (2.5.12)
Reference solution (c) Dilute 1 mL of reference solution (a) M.aximum 10 per cent) determined on 0.300 g.
to 10 mL with methanolR. Sulfuted ash (2.4.14)
Coluitm: Maximum 0.1 per cent) determined on 1.0 g.
- size: 1= 0.15 m, 0 4.6 mm; = ASSAY
- stationary phase: end-eapped o<tade<y/si/y/ silica gelfor
chromawgraphy R (5 pm); Dissolve 2.00 g in water R and dilute to 100.0 mL with the
- temperature: 30 "C. same solvent. Transfer 25.0 mL of the solution to a
separating funnel, add 25 mL of methylene chloride R, 10 mL
Mobile phase:
of 0.1 M sodium hydroxide and 10.0 mLofa freshly prepared
- mobile phaseA: dissolve 1.09 g of sodium hexanesulfonau R
50 gIL solution of potassium Wide R. Shake well) aUow to
and 6.9 g of sodium dihydrogen phosphate monohydrate R in
separate and discard the methylene chloride layer. Shake the
waterfor chromawgraphy R; adjust to pH 3.5 with
aqueous layer with 3 quantities, each of 10 mL, of methylene
phosphan'c acidR and dilute to 1000 mL with the same
chloride R and discard the methylene chloride layers. To the
solvent,
aqueous layer add 40 mL of hydrochloric add R) allow to cool
- mobile phaseB: methanol R2,
and titrate with 0.05 M potassium iodate until the deep-brown
colour is almost discharged. Add 5 mL of methylene chloride R
Thn. Mobile phase A Mobne phase B
(min) (per cent VIJ.') (per cent VM
and continue the titration, shaking vigorously, until the
methylene chloride layer no longer changes colour. Carry out
0-10 SO 20
a blank titration on a mixture of 10.0 mL of the freshly
10·14 80 ---> 50 20 ---> 50
prepared 50 gIL solution of patassium iodide R, 20 mL of
14·35 50 50
water R and 40 mL of hydnxhloric acid R.
35 ~ 36 50 ..... 20 50 ..... 80
36 - 55 20 80
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2022 Benzalkonium Chloride 1-275
1 mL of 0.05 M potassium iodate is equivalent to 1~ mg of Results The principal peaks in the chromatogram obtained
benzalkonium chloride where x is the average relative
principal peaks in the chromatogram obtainedwith the
molecular mass of the sample.
reference solution.
STORAGE C. To 0.05 mL add 2 mL of waterR, 0.1 mL of glacial acetic
In an airtight container. acid Rand) dropwise, I mL of sodium tetraphenylborate
IMPURITIES solution R. A white precipitate is formed. Filter. Dissolve the
Specified impurities A, B, C. precipitate in a mixture of 1 mL of acetone R and 5 mL of
ethanol (96 per "'til) R, heating to not more than 70 "C.
Add water R dropwise to the warm solutionuntil a slight
opalescence forms. Heat gently until the solution is clear and
aUow to cool. White crystals separate. Filter) wash with
3 quantities, each of 10 ml., of water R and dry in vacuo
A. benzyl alcohol, (2.2.32) at a temperature not exceeding 50°C. The crystals
melt (2.2.14) at 127 "C to 133 "C.
('1 D. To 5 mL of dilute sodium hydroxide solution R add 0.1 mL
~CHO of bromophenol bluesolution Rl and 5 mL of methylme
chloride R and shake. The methylene chloride layeris
B. benzaldehyde, colourless. Add 0.05 mL of the solutionto be examined and
shake. The methylene chlorid~ layer becomes blue.
E. To 0.05 mL add I mL of dilute nitric acidR. A white
precipitate is formed whichdissolves on the addition of 5 mL
of ethanol (96 per cenV R. The solution gives reaction (a) of
C. (cWoromethyl)benzene. chlorides (2.3.1).
__________________ ~~ PIlE" TESTS
Solution S
Dilute 2.0 g to 100 mL with carbon dioxide-free waterR.
Benzalkonium Chloride Solution ***
*** ***
than reference solution Y6 (2.2.2, Method If).
(ph. Bur. monograph 0371) *** Acidity or alkal1nlty
Action and use To 50 mL of solution S add 0.1 mL of bromocresol purple
Antiseptic. solution R. Not more than 0.1 mL of 0.1 M hydrochlonc acid
PIlE" _
or 0.1 M sodium hydroxide is required to change the colour of
the indicator.
DEFINITION Average relative molecular mass and ratio of alkyl
Aqueous solution of a mixture of components
alkylbenzyldimethylammonium chlorides, the alkyl groups liquid chromatography (2.2.29).
mainly having chain lengths of C l 2J C14 and CJ6 • Test solution Determine the density (2.2.5) of the solution to
Content be examined. Dilute a quantity of the solution [0 be
475 gIL to 525 gIL of alkylbenzyldimethylammonium examined equivalent to about 0.400 g of benzalkonium
chlorides, calculated using the average relative molecular chloride to 100.0 mL with waterR.
mass (see Tests). The solution may contain ethanol Reference solution Dissolve the contents of a vial of
(96 per cent). benzalhonium chloride for system suitability CRS in 5 mL of
CHARACTERS water R.
Appearance Column:
Clear, colourless or slightly yellowish liquid. - size: / = 0.25 m) 0 = 4.6 mm;
Solubility - srationary phase: end-copped cyanosilyl silica gelfor
Miscible with water and with ethanol (96 per cent). chromaUJgraphy R (5 urn).
It froths copiously when shaken. Mobile phase Mix 45 volumes of aceUJnitrile Rand
55 volwnes of a 13.6 gIL solution of sodium acetate R
IDENTIFICATION previously adjusted to pH 5.0 with glacial acetic acid R.
First identification: B, E. Flow rate 2.0 mllmin.
Second identification: A.. C, D, E. Delation Spectrophotometer at 254 om.
Injection I0 ~L.
(2.2.25).
Identification of homdogues Use the chromatogram supplied
Test solution Dilute 0.3 mL to 100.0 mL with water R.
with benzalkcnium chloride for system suitability CRS and the
Spectral range 220-350 nm. chromatogram obtained with the reference solution to
Absorption maxima At 257 DID) 263 run and 269 run. identify the peaks due to homologues Cia, C I4 and C J6 .
Shoulder At about 250 nm. Relative retention With reference to CJ2 homologue
B. Examine the chromatograms obtainedin the test for (retention time = about 6 min): C14 homologue = about 1.3;
average relative molecular mass and ratio of alkyl C16 homologue = about 1.7.
components.
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1-276 Benzalkonium Chloride 2022
System suitability Reference solution: FIooJ rate 1.0 mUmin.

- resolution: minimum 1.5 between the peaks due to the C 12 Detection Spectrophotometer at 210 nm for impurities A
and C l 4 homologues. and C, and at 257 nm for impurity B.
Calculate the average relative molecular mass of the sample Injection 20 ~L
by summing the products for each homologue, using the
Relative retention With reference to impurity A (retention
impurity C = about 2.4.
w(~) System suitabiliry At 210 run:
A area of the peak due 10 the given homologue in the
chromatognun obtained with the test solution; the chromatogram obtained with reference solution (c);
B sum of the areas of the peaksdue (0 all homologues in the - symmetry factor: minimum 0.6 for the peak due to
chromatogram obtained with me test soIutionj impurity A in the chromatogram obtained with reference
W relative molecular mass for the given hom,ologue: 340, 368 and solution (a).
396 for the C 1b C 14 and C l 6 homologues, respectively.
Limits:
Calculate the percentage of each homologue) using the - correction factor: for the calculation of content, multiply the
following expression: peak area of impurity C by 1.3;
- impurity A: not more than the area of the corresponding
C product of the relative molecular mass of the given homologue
- impun·ty B: not more than the area of the corresponding
and the area of the com:sponding peak in me chromatogram peak in the chromatogram obtained with reference
obtained with the ten solution; solution (h) (0.15 per cent);
D sum of the C values for aU homologues quantified. - impurity C: not more than 0.1 times the area of the
Limits: reference solution (a> (0.05 per cent).
- C I Z homologue: minimum 40 per cent;
- C14 homologue: minimum 20 per cent; Amines and amine salts
- sum of C 12 and C 14 homo/ogues: minimum 70 per cent Mix 10.0 g, while heating, with 20 mL of a mixture of
3 volumes of 1 M hydro<hloric acid and 97 volumes of
Impurities A, Band C methanol R and add 100 mL of 2-propanol R. Pass a stream
liquid chromatography (2.2.29). Prepare the solutions of nitrogen R slowly through the solution. Titrate with up to
immediatelY before use. . 12.0 mL of 0.1 M 'etrabury/ammonium hydroxUk and record
TeJt solution Determine the density (2.2.5) of the solution to the potentiometric titration curve (2.2.20). If the curve shows
be examined. Dilute a quantity of the solution to be 2 points of inflexion, the volume of titrant added between the
examined equivalent to 2.5 g of benzalkonium chloride to 2 points is not greater than 5.0 mL. If the curve shows no
50.0 mL with methanol R. point of inflexion, the solution to be examined does not
Reference soluMn (a) Dissolve 25.0 mg of benzylalcohol CRS comply with the test. If the curve shows I point of inflexion,
(impurity A) in methanol R and dilute to 100.0 mL with the repeat the test but add 3.0 mL of a 25.0 gIL solution of
same solvent. dimethylde<y/amine R in 2-propa",,1 R before the titration.
Reference solution (b) Dissolve 75.0 mg of benzaldehyde CRS If the titration curve after the addition of 12.0 roL of the
(impurity B) in methanol R and dilute to 100.0 mL with the titrant shows only 1 point of inflexion, the solution to be
same solvent. Dilute 1.0 mL of this solution to 10.0 mL with examined does not comply with the test.
methanol R. Sulfated ash (2.4.14)
Reference solution (e) Dilute I mL of reference solution (a) Maximwn 0.1 per cent, determined on 1.0 g.
to 10 mL with methanol R. ASSAY
Column: Determine the density (2.2.5) of the solution to be examined.
- size: 1= 0.15 m, 0 4.6 mm; = Dilute 4.00 g to 100.0 mL with water R. Transfer 25.0 mL
- staticnary phase: end-capped octmk<y/si(yl silica gelfor of the solution to a separating funnel, add 25 mL of
chromawgraphy R (5 urn), methylene chloride R, 10 mL of 0.1 M sodium hydroxide and
- temperature: 30 "C. 10.0 mLofa freshly prepared 50 gIL solution of patossium
Mobile phase: iodide R. Shake well, allow to separate and discard the
- mobile phase A: dissolve 1.09 g of sodium hexanesu/fonate R methylene chloride layer. Shake the aqueous layer with
and 6.9 g of sodium dihydrogen phosphate monohydrate R in 3 quantities, each of 10 mL, of methy/em chloride R and
waterfor chromatography R; adjust to pH 3.5 with discard the methylene chloride layers. To the aqueous layer
phosphoric acidR and dilute to 1000 mL with the same add 40 mL of hydrochloric acidR, aUow to cool and titrate
solvent; with 0.05 M potassium iodate until the deep-brown colour is
- mobile phaseB: methanol R2; almost discharged. Add 5 mL of methyl"" chloride Rand
continue the titration, shaking vigorously, until the methylene
Time MobUe phase A MobUe phase B chloride layer no longer changes colour. Carry out a blank
(mlo> (per cent V/V) (per cent VIP)
titration on a mixture of 10.0 mL of the freshly prepared
0·10 SO 20 50 gIL solution of potassium iodide R, 20 mL of water Rand
10 - 14 80 -> 50 20 -> 50 40 mL of hydrochloric acidR.
14 - 35 50 50
50 ..... 80
I mL of 0.05 M potassium iodate is equivalent to I~ mg of
35 - 36 50 -. 20
36 455
20 SO benzalkonium chloride where x is the average relative
molecular mass of the sample.
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2022 Benzathine Benzylpenicillin 1-277
LABELLING Solubility
The label states the content of ethanol (96 per cent), if any. Very slightly soluble in water, freely soluble in
dimethylfonnamide and in formamide, slightly soluble in
IMPURITIES
ethanol (96 per cent).
Specified impurities A.. B~ C.
IDENTIFICATION
A. benzyl alcohol, Comparison benzathine benzylpenicillin CRS.
r-
~CHO
examined in 5 mL of methanol R.
Reference solution Dissolve 25 mg of benzathine
benzylpenicillin CRS in 5 mL of methanol R.
B. benzaldehyde,
Plate TLC silanised silica gelplate R.
tl'[obile phase i\tlix 30 volumes of acewne Rand 70 volumes
of a 154 gIL solution of ammonium Gatate R previously
adjusted to pH 7.0 with ammonia R.
Application I ~L.
C. (chloromethyl)benzene.
Deoelopmem Over 2/3 of the plate.
_ _ _ _ _ _~ PhE"
Drying In air.
Detection Expose to iodine vapour until the spots appear
Benzathine Benzylpenicillin System suitability Reference solution:
Tetrahydrate Results The 2 principal spots in the chromatogram obtained
Benzathine Benzylpenicillin with the test solution are similar in position, colour and size
(Benzy/peniciUin (Benzarhine) Tetrahydnue, Ph. Bur. to the 2 principal spots in the chromatogram obtained with
manograph 0373) the reference solution.
C. Place about 2 mg in a test-tube about 150 mm long and
15 mm in diameter. Moisten with 0.05 mL of water Rand
add 2 mL of sulfuric acid-formaldehyde reagenl R. Mix the
contents of the tube by swirling; the solution is practically
4H,o
colourless. Place the test-tube on a water-bath for 1 min;
a reddish-brown colour develops.
D. To 0.1 g add 2 mL of 1 M sodimn hydroxide and shake for
2 min. Shake the mixture with 2 quantities, each of 3 mL, of
981 41372-02-5 ether R. Evaporate the combined ether layers to dryness and
dissolve the residue in I mL of ethanol (50 per ernl VII1 R.
Action and use Add 5 mL of picric acidsolution R, heat at 90 "C for 5 min
Penicillin antibacterial. and allow to cool slowly. Separate the crystals and
PhEII _ recrystallise from ethanol (25 per cent VllQ R containing
10 gIL of picric acid R. The crystals melt (2.2.14) at about
DEFINITION 214 "C.
Nt,N' -Dibenzylethane-I,2-diamine bis[(2S,5R,6R)-3,3- TESTS
dimethyl- 7-oxo- 6-(2-phenylacetamido)-4-thia-I-a2abicyclo
Acidity or 3Ikal1nlty
[3. 2.0jheptane-2-carboxylatej tetrahydrate.
To 0.50 g add 100 mL of carbon dioxide-free water Rand
Salt obtained from Benzylpenicillin sodimn (0114) or shake for 5 min. Filter through a sintered-glass filter (2.1.2).
Benzylpenicillin potassium (0113) produced by the growth of To 20 mL of the filtrate add 0.1 mL of bromothymol blue
certain strains of Penicillium nOlalutti or related micro- solution RI. The solution is green or yellow. Not more than
organisms. 0.2 mL of 0.02 M sodium hydroxide is required to change the
Content colour of the indicator to blue.
- benzathine benzylpenidJlin: 94.5 per cent to 102.0 per cent Related substances
(anhydrous substance) without correction for dispersing or Liquid chromatography (2.2.29). Prepare the solutions
suspending agents; immediately before use and by diluting to volume immediately
- benzathine: 24.0 per cent to 27.0 per cent (anhydrous afterdissolution.
substance).
Solution A Prepare a solution containing 1.3 gIL of disodium
Dispersing or suspending agents (e.g. lecithin and hydrogen phosphate dodecahydrate Rand 6.8 gIL of potassimn
polysorbate 80) may be added. dihydrogen phosphate R.
CHARACTERS Test solution (a) Dissolve 40.0 mg of the substance to be
Appearance examined in 50 mL of methanol R and dilute to 100.0 mL
White or almost white, slightly hygroscopic powder. with solution A.
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1-278 Benzathine Benzylpenicillin 2022
Test solution (b) Dissolve 70.0 mg of the substance to be Calculation of percentag« etmtents:
examined in 25 mL of methanol R and dilute to 50.0 mL - correction factors: multiply the peak areas of the following
with solution A. impurities by the corresponding correction factor:
Reference solution (a) Dissolve 40.0 mg of benzathine impurity E = 1.9; impurity F = 1.5;
benzylpenic~/in CRS in 50 mL of methanol R and dilute to - for each impurity, use me concentration of
100.0 mL with solution A. '0
benzylpenicilUn reference solution (c).
Reference solution (b) Dissolve 3 mg of benzathine Limits:
benzylpenicillin for peak idenrijication CRS (containing - impurily C: maximum 2.0 per cent;
impwities A, BJ C, D, E, F, G, H, I, J and K) in 1 mL of - impun·ty K: maximum 1.0 per cent;
methanol R and dilute to 2 mL with solution A. - ~"mpuniy J: maximum 0.5 per cent;
Reference solurion (c) Dilute 1.0 mL of reference solution (a) - impurilits E (sum of isomers), F (sum of epimerr): for each
£0 20.0 mL with a mixture of equal volumes of methanol R
impurity, maximum 0.3 per cent;
and solution A. - impun'ties A, B, D, G, H, I: for each impurity, maximum
0.2 per cent;
Reference solution (d) Dilute 3.0 mL of reference solution (c) - any other impun·,y: for each impurity, maximum
to 100.0 mL with a mixture of equal volumes of methanol R 0.2 per cent;
and solution A. - total: maximum 3.5 per cent;
Column: - reporting threshold: 0.05 per cent; disregard the peak due (0
- size: I = 0.15 m, 0 = 4.6 nun; benzathine.
- stationary phase: end-capped acradecylsilyl silica gelfor
Water (2.5.12)
chromalography R (3 urn);
Bacterial endotoxin. (2.6.14, Method E)
Mobile phase:
Less than 0.13 lU/mL, if intended for use in the
- mobile phase A: mix 10 volumes of a 34 gIL solution of
potassium dihydrogen phosphate R previously adjusted to manufacture of parenteral preparations without a further
pH 3.3 with phosphoric acid R, 30 volumes of methanol RI
appropriate procedure for the removal of bacterial
and 60 volumes of waterfor chromatography Rj
endotoxins.
- mobile phase B: mix 5 volumes of a 34 gIL solution of Suspend 20 mg in 20 mL of a solutionof 0.1 ..IH sodium
potassium dihydrogen phosphate R previously adjusted to hydroxide diluted 1 to 100, shake thoroughly and centrifuge.
pH 3.3 with phosphoric acidR, 25 volumes of warer far Examine me supernatant.
chromatography Rand 7Q volumes of methanol RI; ASSAY
Tim, MobUe phase A Moblle phase B related substances with the following modifications.
(min) (per cent PIJ? (per cent Vm
Mobile phase Mobile phase B. mobile phase A (15:85 VIV).
0-2 85 15
2· ]6 85 ..... 0 15 ..... 100 Injection Test solution (a) and reference solution(a).
16·26 0 100 System suitability Reference solution (a):
Flow rate 1.5 mllmin. benzylpenicillin.
Detection Spectrophotometer at 220 run. Calculate the percentage contents ofbenzathine (C I6H2 oN'2)
and benzarhine benzylpenicillin (C.8H,oN.08S,) taking into
Injection 20 J1L of test solution (b) and reference account the assigned content of benzathine
benzylpenicillin CRS.
with benzathine benzylpenicillin far peak identijication CRS and STORAGE
the chromatogram obtained with reference solution (b) {Q In an airtight container, If the substance is sterile, the
identify the peaks due to impurities A, B, C, D, E, F, GJ H, container is also sterile and tamper-evident.
1, J and K. IMPURITIES
Relative retention With reference to benzylpenicillin Specified imPUri.,ies A, B, C, D, E, F, G, H, I, J, K.
(retention time :::;; about 7 min): impurity A := about 0.18,;
benzathine = about 0.30; impurity D ;:;; about 0.36,;
impurity G = about 0.38; impurity J = about 0.44;
= =
impurity E about 0.51 and 0.60; impurity B about 0.69;
= =
impurity F about 0.84 and 0.88; impurity H about 1.22;
impurity I = about 1.42; impurity C = about 1.75; A. N I-benzylethane-l,2-diamine,
=
impurity K about 2.90.
System suitability:
- resolution: minimum 1.0 between the peaks due to the
eplmers of impurity F and minimum 1.5 between the
peaks due to impurities D and G in the chromatogram B. phenylacetic acid,
obtained with referencesolution (b);
the chromatogram obtained with reference solution (d).
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2022 Benzatropine Mesilate 1-279
o)-~~~:,
H3C~~--H--s CH3
II H H
o
I. (2S,5R,6R)-6-hexanamido-3,3-dimethyl-7-oxo-4-thia-l-
(dihydropenicillin F),
J. unknown structure,
C. (23,4S)-2-[(13)-2-[benzyl[2-(benzylamino)ethyl]amino]-
2-oxo-l-(2-phenylacetamido)ethyl]-5,5-dimethyl-1,3-
thiazolidine-4-carboxylic acid (benzylpenicilloic acids
benzathide),
D. (3S,7R,7aR)-5-benzyl-2,2-dimethyl-2,3,7,7a-
tetrahydroimidaw[5, I-b] [1,3]thiazole-3,7-dicarboxylic acid
(penillic acid of benzylpenicillin), . K. (2R,2'R,4S,4' S)-2,2'-[(4R,IIRJ-6,9-dibenzyl-2,5,10,13-
tetraoxo-l,14-diphenyl-3,6,9,12-tetraazatetradecane-4J 11-
diyl]bis(5,5-dimethyl-I,3-thiazolidine-4-carboxylic acid).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEor
E. (23,4S)-2-[(3)-carboxy(2-phenylacetamido)methyl]-5,5- Benzatropine Mesilate

dimethyl-I,3-thiawlidine-4-carboxylic acid (penicilloic
acids of benzylpenicillin), Me
I
;t\
VyH ,CH,SO,H
OCHPh 2
403.5 132-17-2
F. (2RS,4S)-5,5-dimethyl-2-[(2-phenylacetamido)methyl]-
1,3-thiazolidine-4-carboxylic acid (penilloic acids of Acdon and use
benzylpenicillin), Anticholinergic.
H
Preparations
o)-~Xc~:, Benzatropine Injection
vr
~- - H--S)<CH3
Benzatropine Tablets
I H H DEFINITION
HO ~ 0 Benzatropine Mesilare is (IR,3R,5S)-3-benzhydryloxytropane
methanesulfonate. It contains not less than 98.0% and not
G. (2S,5R,6R)-6-[2-(4-hydroxyphenyl)acetamido]-3,3- more than 100.5% of C21H2SNO,CH403S, calculated with
dimethyl-7-oxo-4-thia-l-azabicyclo[3.2.0]heptane-2- reference to the dried substance.
carboxylic acid,
PRODUCTION
H Risk assessmentshould be used to evaluate the potential for
H'C~ O)-~XC~: genotoxic methanesulfonate esters to be. formed in the
presence of low molecular weight alcohols. If a riskof
·~····H--S)<CH: methanesulfonate ester formation is identified through risk
H H assessment, these impurities should not exceed the threshold
o
of toxicological concern.
H. (2S,5R,6R)-6-[(3Z)-hex-3-enamido]-3,3-dimethyl-7-oxo- CHARACTERISTICS
4-thia-l-azabicyclo[3.2.0]heptane-2-carboxylic acid A white, crystalline powder. It melts at about 144°.
(isopenicillin F), Very soluble in water; freely soluble in ethanol (96%);
practically insoluble in ether.
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1-280 Benzbromarone 2022
IDENTIFICATION Time Mobile phase A Mobile phase B Commenl

A. Dry the substance at 105 0 for 3 hours. The infrared (Minutes) (·4 vlv) (% v/V)
absorption spectrum, Appendix II A, is concordant with the 0-20 70---)30 30---)70 lioeargradienl
reference spectrum of benzatropine mesilate (RS 026). 20-30 3O~O 70---)100 ~near gradient
B. The light absorption, Appendix II B, in me range 230 to
3D-55 0 100 lsocrabc
350 nm ofa 0.1% wlv solution in 2M hydrodJloric acid
55-05 70 30 lsocr"atic
exhibits two maxima, at 253 and 258 om. The absorbance at
253 run is about 0.96 and at 258 run is about 1.1.
C. Dissolve 10 mg in 2 mL of water, pour into 5 mL of hot Inject 20 ~L of solution (4). The test is not valid unless the
picric acid solution Rl and allow to cool. The melting pointof resolution factor between the two principal peaksis at least I.
the precipitate, after drying at 105°, is about 185°, If necessary adjust the concentration of acetonitrile or adjust
Appendix V A. the time program of the linear gradient elution.
TESTS Inject separately 20 Il1. of mobile phase A as a blank and
Tropine 20 ~L each of solutions (I), (2) and (3). In the
Carry out the method for thin-layer chromarography, chromatogram obtainedwith solution (1) the area of any
Appendix III A, using silica gel G as the coating substance peak corresponding to desmethyl benzatropine is not greater
and a mixture of 75 volumes of e/hanol (96%) and than the area of the principal peak in the chromatogram
15 volumes of 13.5M ammonia as the mobile phase. Apply obtained with solution (3) (0.5%), the area of any other
separately to the plate 10 J.1L of each of two solutions in secondary peak is not greater that the area of the principal
acetone containing (1) 4.0% wlv of the substance being peak in the chromatogram obtained with solution (2) (0.2%)
examined and (2) 0.020% wlv of tropine. Afterremoval of the and the stun of the areas of any such peaks is not greater
plate, allow it to dry in air and spray with sodium than 2.5 times the area Ofthe principal peak in the
iodoWmumate solution and then with a 0.4% wlv solutionof chromatogram obtained with solution (2) (0.5%). In solution
sulfuric acid. Any spot corresponding 10 tropine in the (I) disregard any peaks corresponding 10 the peaks in the
chromatogram obtained with solution (1) is not more intense chromatogram obtainedwith the blanksolution.
than the spot in the chromatogram obtained with Loss on drying
solution (2). When dried to constant weightat 105°, loses not more than
Related substances 5.0% of its weight. Use I g.
Carry out the method for liquidchromawgraphy, Sulfated ash
Appendix ill D J using the following solutions. For solution Not more than 0.1%, Appendix IXA.
(I) mix with the aid of ultrasound 50 mg of the substance
ASSAY
being examined with 15 mL of mobile phaseA, dilute to
Dissolve 0.6 g in 25 mL of water, add 5 mL of d.lmesadium
50 mL with the same solvent and filter. For solution(2)
carbonate solution and extract with four 10 mL quantities of
dilute 1 volume of solution (1) to 100 volumeswith mobile
chlorcform. Wash the combined extracts with 10 mL of water,
phase A and further dilute 1 volumeof the resulting solution
extract the washings with 5 mL of chlorofonn and add the
to 5 volumeswith the same solvent. For solution (3) mix
chloroform to the combined extracts. Filter and wash the
with the aid of ultrasound 50 mg of tksmethyl bensatropine
filter with 5 mL of chlorofonn. To the combined filtrate and
hydrochkJride BPCRS with 15 mL of mobile phase A, dilute
washings add 25 mL of 1,4-dioxan and titrate with O.lM
to 100 mL and dilute 1 volume of the resulting solution to
perchloric acid VS using 0.15 mL of a 0.1% wlv solution of
100 volwnes with the same solvent. Solution (4) contains
methyl red in methanol as indicator. Each mL of O.IM
0.01% wlv each of benzatropine mesikue BPCRS and desme/hy/
perchloric acid VS is equivalent to 40.35 mg of C 2IH2,NO,
benzatrapine hydrochlaride BPCRS in mobile phase A.
CH.O,S.
The chromatographic procedure may be carried out using
(a) a stainless steel column (25 em x 4.6 mm) packed with
phenylsi/y/ silica gelfor chromatagraphy (5 urn) (Zotb ax
SB-Phenyl 5~ is suitable). Carry out a linear gradient elution
with a flow rate of 1 mL per minute using the following
Benzbromarone
conditions. Use a detection wavelength of 220 run. (ph. Bur. monograph 1393)
Mobile phase A A mixture of 5 volumes of a 1M potassium
phosphate buffer prepared as described for mobile phase B, H,C
20 volumesof cuetonitrile and 75 volumes of water. Br
Mobile phaseB A mixture of 35 volumesof water,
60 volumes of lKetqnilrile and 5 volumes of a 1M potassium OH
phosphate buffer prepared in the following manner: dissolve
136.1 g of potassium dihydrogen ortiwphosphare in 900 mL of
water, add 5 mL of anhaphosphoric acid (85%) and dilute to
1000 mL. 424.1 3562-84-3
Action and use

Uricosuric; treatment of hyperuricaemia.
PhE" _
DEFINITION
(3,5-Dibromo-4-hydroxyphenyl)(2-ethylbenzofuran-3-
yl)methanone.
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2022 Benzbromarone 1-281
Content - impurity B: not more than 10 times the area of the

98.0 per cent to 101.0 per cent (dried substance). principal peak in the chromatogram obtained with
reference solution (a) (1.0 percent);
CHARACTERS
Appearance
with reference solution (a) (0.10 percent);
Solubillty - sum of impurities other thanA and B: not more than twice
Practically insoluble in water, freely soluble in acetone and in the area of the principal peak ln the chromatogram
methylene chloride, sparingly soluble in ethanol obtained with reference solution (a) (0.2 per cent);
(96 per cent). - disregard limir. 0.2 times the area of the principal peak in
mp the chromatogram obtained with reference solution (a)
About 152 ·C. (0.02 per cent).
IDENTIFICATION Halides expressed as cblorides (2.4.4)
A. Infrared absorption spectrophotometry (2.2.24). Maximum 400 ppm.
Comparison benebromarone CRS. Shake 1.25 g with a mixture of 5 mL of dilute nitric acid R
and 15 mL of water R. Filter. Rinse the filter with water R
B. By means of a copper wire, previously ignited, introducea
and dilute the filtrate to 25 mL with the same solvent. Dilute
smaU amount of the substance to be examined into the 000-
2.5 mL of this solution to 15 mL with water R.
luminous part of a flame. The colourof the flame becomes
green. Iron (2.4.9)
Maximum 125 ppm.
TESTS
Moisten the residue obtained in the test for sulfated ash with
2 mL of hydrochlori< acidR and evaporate to dryness on a
water-bath. Add 0.05 mL of hydrochlori< acid Rand 10 mL of
than referencesolution Y5 (2.2.2, Metlwd II).
waur R, heat [Q boiling and maintain boilingfor 1 min.
Dissolve 1.25 g in dimethylformamide R and dilute to 25 mL Allowto cool. Rinse the crucible with water R, collect the
with the same solvent. rinsings and dilute to 25 mL with water R. Dilute 2 mL of
Acldity or alka1lnlty this solution to 10 mL with water R.
Shake 0.5 g with 10 mL of carbon dioxide-free waterR for Loss on drying (2.2.32)
1 min and filter. To 2.0 mL of the filtrate add 0.1 mL of Maximum 0.5 per cent, determined on 1.000 g by drying in
methyl red soluaon Rand 0.1 mL of 0.01 M hydrochlori< acid. vacuo at 50 ·C for 4 h.
The solution is red. Add·0.3 mL of 0.01 M sodium hydroxide.
The solution is yellow. Sulfated ash (2.4.14)
Related substances
Liquid chromatography (2.2.29). ASSAY
Test solution Dissolve 0.125 g of the substance to be Dissolve 0.300 g in 60 mL of methanol R. Stir until
examined in 30 mL of methanol R and dilute to 50.0 mL completely dissolved and add 10 mL of water R. Titrate with
with the mobile phase. 0.1 M sodium hydroxide, determining the end-point
100.0 mL with the mobile phase. Dilute 1.0 mL of this 1 mL of 0.1 M sodium hydroxide is equivalent to 42.41 mg
solution to 10.0 mL with the mobile phase. of CI7H12Br203.
Reference solution (b) Dissolve 10 mg of benzarone CRS STORAGE
(impurity C) in the mobile phase and dilute to 20 mL with Protected from light.
the mobile phase. To 5 mL of this solution add I mL of the IMPURITffiS
test solutionand dilute (0 100 mL with the mobile phase. Specified impurities A, B.
Column: Other detectable impun'ties (thefollowing substances would, if
- size: 1= 0.25 m, {2} = 4.6 rom; present at a sufficient level, be detected by oneor other of the tests
- stationary phase: ocrodecylsilyl ,ilicagelfor chromatography R in the monograph. They are limited by thegeneral acaptance
(5 um). criterion for otherhmspecified impurities and/orby the general
Mobile phase glacial acetic acid R, acetonitrile R, waterR, monograph Substances for pharmaceutical use (2034). It is
methanol R (5:25:300:990 VIVIVIV). there/ore not neassary to idemify these impun·ties for
Flow rate 1.5 mUmin. demonstration of compliance. See also 5.10. Control of impurities
Detection Spectrophotometer at 231 om. in substances for pharmaceutical use) C.
Injecu'on 20 1l1..
Run time 2.5 times the retention time of benzbromarone.
Relative retention With reference to benzbromarone:
impurity A = about 0.6; impurity B = about 2. OH
System suitability Reference solution (b): B,
impurity C (I" peak) and benzbromarone (2'd peak).
A. (3-bromo-4-hydroxyphenyl)(2-ethylbenzofuran-3-yl)
Limits: methanone,
- impurity A: not more than 4 times the area of the
reference solution (a) (0.4 percent);
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1-282 Benzethonium Chloride 2022
'" Drying In a current of warm air.

Detection Examine in ultraviolet lightat 254 nm.
OH
B,
with the test solution is similar in position andsize to the
B' principal spot in the chromatogram obtained with the
reference solution.
B. (6-bromo-2-ethylbenzofuran-3-yl)(3,5-dibromo-4-
C. To 5 mL of diiutesodium hydroxide sol"tion R add 0.1 mL
hydroxyphenyljmethanone,
of bromophenol blue sduuon Rl and 5 mL of methylene
H'C~O
chloride R and shake. The lowerlayer is colourless.
Add 0.1 mL of solution S (see Tests) and shake. A blue
o I colour develops in the lowerlayer.
::0- '::::::,.. OH D. To 2 mL of solution S add 1 mL of di/"w nitric acid R.
A white precipitate is formed which dissolves upon addition
'- of 5 mL of ethanol (96 percenO R. The solution gives
C. (2-ethylbenzofuran-3-yl)(4-hydroxyphenyl)methanone reaction (a) of chlorides (2.3.1).
(benzarone). TESTS
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhElI
Solution S
Benzethonium Chloride Appearance of solution
Solution S is clear(2.2.1) and not more intensely coloured
(ph. Bur. monograph 0974) than reference solution Y. (2.2.2, Method II).
To 25 mL of solution S add 0.1 mL of phenolphtholein
sohuion R. The solution is colourless. Add 0.3 mL of 0.01 M
sodium hydroxide. The solution is pink. Add 0.1 mL of methyl
red sol"tion Rand 0.5 mL of 0.01 M hydrochkni< acid.
The solution is orange-red.
Volatile bases and salts of volatile bases (2.4.1,
448.1 121-54-0 Method B)
Maximum 50 ppm, determined on 0.20 g.
Action and use Prepare the standard using 0.1 mL of ammonium standard
Antiseptic. sollll;"n (100 ppm lifH.,) R. Replace heavy magnesium oxide
PhElI _ by 2.0 mL of strong sodium hydroxide solution R.
DEFINITION
lif-Benzyl-lif~-dimethyl-2-[2-[4-(I,I,3,3
tetramethylbutyl)phenoxy]ethoxy]ethanaminium chloride.
Content
ASSAY
CHARACTERS
Dissolve 2.000 g in wawr R and dilute to 100.0 mL with the
Appearance
same solvent. Transfer 25.0 mL of the solution to a
White or yellowish-white powder.
separating funnel, add 10 mL of a 4 gIL solution of sodium
Solubility hydroxide R, 10.0 mL of a freshly prepared 50 gIL solution of
Very soluble in water and in ethanol (96 per cent), freely potassium iodide Rand 25 mL of methylene chloride R. Shake
soluble in methylene chloride. vigorously, allow to separate and discard the lower layer.
An aqueous solution froths copiously whenshaken. Shake me upperlayer with 3 quantities, each of 10 ml., of
IDENTIFICATION methylene chloride R and discard the lower layers. To the
A. Melting point (2.2.14): 158°C to 164 °C, after drying at upper layer add 40 mL of hydrochloric acid R, allow to cool
105°C for 4 h. and titrate with 0.05 M potassium iodate until the deep brown
colour is almost discharged. Add 4 mL of methylene chloride R
B. TItin-Iayer chromatography (2.2.27).
and continue the titration, shaking vigorously, until the lower
layer is no longer brown. Carry out a blank titration using a
mixture of 10.0 mL of a freshly prepared 50 gIL solution of
solvent.
potassium iodide R, 20 mL of water Rand 40 mL of
Reference soluuon Dissolve 25 mg of benzethonium hydrochloric acid R.
chloride CRS in water R and dilute to 5 mL with the same
I mL of 0.05 M potassium iodate is equivalent to 44.81 mg of
solvent.
C"H,2 CIN02'
Plaw TLC silica gel F254 plow R.
STORAGE
Mobile phase glacial acetic acid R, water RJ methanol R
(5:5:100 VIVIV).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEll
Application 20 ~L.
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2022 Benzocaine 1-283
Column:
Benzocaine - size: I;;;; 0.10 m, (2) = 4"6 mm;
(Ph. Bur. monograph 0011) - stationary phase: end-capped oaadecylsi(yl silica gelfor
chromatography compatible with 100 per centaqueous mobile
phases R (3 pm);
MoMe phase:
- mobile phase A: dilute I mL ofperchloric acid R to 100 mL
with waterfor chromatography Rj dilute 1 mL of this
solution to 100 mL with water for chromatography R; mix
165.2 94-09-7 9 volumes of this solution and I volume of acetonitrile Rlj
- mobile phaseB: acetonitrile Rl;
Action and use
Local anaesthetic.
Time MobUe phase A Moblle phase B
Ph'" _ (min) (percent VII? (per cent V/l?
0-2 100 o
DEFINITION 2 - 15 100 --> 38.5 o --> 61.5
Emy} 4-aminobenzoate.
Content Flow rate 1.5 mUmin.
CHARACTERS Injection I0 ~L.
Appearance Identification of impurities Use the chromatogram obtained
White or almost white, crystalline powder or colourless with reference solution (b) to identify the peak due to
crystals. impurity E.
Solubl1lly RekJtwe retention With reference to benzocaine (retention
Very slightly soluble in water, freely soluble in ethanol time ;;;; about 10 min): impurity E = about 0.9"
(96 per cent).
It shows polymorphism (5.9). - resolution: minimum 5.0 between the peaks due to
IDENTIFICATION impurity E and benzocaine.
First identification: A. Calculation of percentage contents:
Second identification: B. - for each impurity, use the concentration of benzocaine in
A. Infrared absorption spectrophotometry (2.2.24). reference solution (a).
Comparison benzocaine CRS. Limits:
0"10 per cent;
substance to he examined and the reference substance
separately in anhydrous ethanol R, evaporate to dryness and - reporting threshold: 0.05 per cent.
B. Melting point (2.2.14).
Maximum 0.5 per cent, detennined on 1.000 g by drying in
Determination A Determine the melting point of the vacuo.
ResultA 89 'C to 92 'C. Maximum 0.1 per cent, determined on 1.0 g.
Determination B Mix equal parts of the substance to be
examined and benzocaine CRS and determine me melting
ASSAY
point of the mixture. Carry out the determination of primary aromatic amino-
nitrogen (2.5"8), using 0.400 g dissolved in a mixture of
Result B The absolute difference between the melting point
25 mL of hydrochloric acid Rand 50 mL of waterR.
of the mixture and the value obtained in determination A is
not greater than 2 "C. 1 mL of 0.1 M sodium nitrue is equivalent to 16.52 mg
of C.H uN02 •
TESTS
Related substances
STORAGE
Liquid chromatography (2.2.29). Protected from light.
Solvent mixture acetonitrile R1, waterfor chromawgraphy R IMPURITIES
(50:50 VIV). Other detectable impurities (thefollowing substances would, if
Test solution Dissolve 25.0 mg of the substance to be present at a sufficient level, be detected by oneor other of the tests
examined in 5 mL of acetonitrile R and dilute to 50.0 mL in the monograph. They are limited by the general acceptance
with the solvent mixture. criterion for otherhmspecified impurities andlor by thegeneral
Reference solution (a) Dilute 1.0 mL of the test solution. to
therefore not neussary to identify these impurities for
demonstration of compliance. See also 5.10. Control of impumies
in substances for phannaceutical use) A, B, C, D, E, F, G, H.
Reference solution (b) Dissolve 5 mg of the subst~ce to ?e
examined and 5 mg of 4-nilrObenzoic acidR (impunty E) m
10 mL of the solvent mixture. Dilute 1 mL of the solution to
50 mL with the solvent mixture.
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1-284 Benzoic Acid 2022
~OH Benzoic Acid

fl,N~
A. (4-aminophenyl)methanol,
(C
I
-..
OH
Nfl,
122.1 65-85-0
B. (2-aminophenyl)methanol,
o Action and use
~o~~,
Antimicrobial preservative.
Preparations
Compound Benzoic Acid Ointment
Nfl, Benzoic Acid Solution
PhE" _
C. ethyl 3-aminobenzoate,
DEFINITION
Benzenecarboxylic acid.
Content
99.0 per cent to 100.5 per cent.
CHARACTERS
D. ethyl 2-aminobenzoate, Appearance
White or almostwhite, crystalline powderor colourless
("yCo,H crystals.
o,N~ Solublllty
Slightly soluble in water, soluble in boilingwater, freely
E. 4-nitrobenzoic acid, soluble in ethanol (96 per cent) and in fatty oils.
IDENTIFICATION
A. Melting point (2.2.14): 121°C to 124 °C.
R Solution S (see Tests) gives reaction (a) of benzoates
(2.3.1).
TESTS
F. (3-aminophenyl)methanol, Solution S
Dissolve 5.0 g in ethonol (96 percen!> R and dilute to
Solution S is clear (2.2.1) and colourless (2.2.2, MethodII).
G. 4-aminobenzoic acid, Carbonisable substances
Dissolve 0.5 g with shaking in 5 mL of sulfuric ocidR. After
5 min, the solution is not more intensely coloured than
reference solution Y5 (2.2.2, Method l).
Oxidisable substances
Dissolve 0.2 g in 10 mL of boiling woter R. Cool, shake and
H. methyl a-aminobenzoate. filter. To the filtrate add I mL of diluu m([uric acidRand
_____ ~ I'hE" 0.2 mL of 0.02 M potassium pennanganace. After 5 min, the
solution is still colouredpink.
Halogenated compounds and halides
Maximum 300 ppm.
AUglassware used must be chloride-flu and may beprepared by
soaking overnight in a 500gIL solution of nitrU acidR, rinsed
with water R and stored full of wale/' R. It is recommended that
glassware be reserved for this res"
Solution (a) Dissolve 6.7 g in a mixture of 40 mL of 1 M
sodium hydroxide and 50 mL of ethanol (96 per<en!> Rand
dilute to 100.0 mL with woter R. To 10.0 mL of this solution
add 7.5 mL of diluu sodium hydroxide solution R and 0.125 g
of nkkel-aluminium alloy R and heat on a water-bath for
10 min. Allow to cool to room temperature) filter into a
25 mL volumetric flask and washwith 3 quantities, each of
2 mL, of ethanol (96 per <en!> R. Dilute the filtrate and
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2022 Hydrous Benzoyl Peroxide 1-285
washings to 25.0 mL with waterR. This solution is used to Solublllty

prepare solution A. Practically insoluble in water, soluble in acetone, soluble in
Solution (b) In the same manner, prepare a similar solution methylene chloride with the separation of water, slightly
without the substance to be examined. This solution is used soluble in ethanol (96 per cent).
to prepare solution B. It loses water rapidly on exposure to air with a risk of
In four 25 mL volumetric flasks, place separately 10 mL of explosion.
solution (a), 10 mL of solution (b), 10 mL of chloride Mix theentire sample thoroughly before carrying out thefollowing
standard solution (8 ppm CD R (used to prepare solution C) tests.
and 10 mL of waterR. To each flask add 5 mL offeme
IDENTIFICATION
ammonium sulfate solution R5, mix and add dropwise and with
First identification: B
swirling 2 mL of nitric add Rand 5 mL of mercuric
thiocyanate solution R. Shake. Dilute the contents of each flask Second identification: A, C, D.
to 25.0 mL with water R and allow the solutions to stand in A. Ultraviolet and visible absorption spectrophotometry
a water-bath at 20°C for 15 min. Measure at 460 run the (2.2.25).
absorbance (2.2.25) of solution A using solution B as the Solution A Dissolve 80.0 mg in ethand (96 per een!! Rand
compensation liquid, and the absorbance of solution C using dilute to 100.0 mL with the same solvent. Dilute 10.0 mL of
the solution obtained with 10 mL of water R as the the solution to 100.0 mL with ethanol (96 per cen!! R.
compensation liquid. The absorbance of solutionA is not Solution B Dilute 10.0 mL of solution A to 100.0 mL with
greater than that of solutionC. ethanol (96 per cen!! R.
Sulfated ash (2.4.14) Spectral ranges 250-300 urn for solution A; 220-250 urn for
Maximwn 0.1 per cent, determined on 1.0 g. solutionB.
ASSAY Absorption maxima At 274 nm for solution A; at 235 nm for
Dissolve 0.200 g in 20 mL of ethanol (96 per een!! Rand solution B.
titrate with 0.1 M sodium hydroxide, using 0.1 mL of phenol Shoulder At about 282 run for solution A.
red solution R as indicator, until the colour changes from Absorbance ratio A 23 s1A274 = 1.17 to 1.21.
yellow to violet-red.
Comparison Ph. Bur. reference spectrum of hydrous benzO)~
C1H60Z '
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
peroxide.
C. Dissolve about 25 mg in 2 mL of acetone R. Add 1 mL of
a 10 gIL solution of diethylphenylenediamine sulfate R and mix.
A red colour develops which quickly darkens and becomes
dark violet within 5 min.
Hydrous Benzoyl Peroxide D. To I g add 5 mL of ethanol (96 per cen!! R, 5 mL of
(ph. Bur. monograph 0704) dilute sodium hydroxide solution R aod 10 mL of waterR. Boil
the mixture underrefluxfor 20 min. Cool. The solution
gives reaction (c) of benzoates (2.3.1).
TESTS
Acldlty
Dissolve a quantity of the substance to be examined
containing the equivalent of 1.0 g of dibenzoyl peroxide in
25 mL of acetone R, add 75 mL of water R and filter. Wash
C,oH'oO. 242.2 94-36-0
the residue with two quantities, each of 10 ml., of water R.
(anhydrous substance)Anhydrousbenzoylperoxide
Combine the filtrate and the washings and add 0.25 mL of
Action and use phenolphthalein solution Rl, Not more than 1.25 mL of 0.1 M
Used topically in the treatment of acne. sodium hydroxide is required to change the colourof the
indicator. Carry out a blanktest.
Preparations
Benzoyl Peroxide and Clindamycin Gel Related substances
Benzoyl Peroxide Cream
Benzoyl Peroxide Gel
Testsolution Dissolve a quantity of the substance to be
Benzoyl Peroxide Lotion examined containing the equivalent of 0.10 g of dibenzoyl
PotassiumHydroxyquinoline Sulfate and Benzoyl Peroxide peroxide in acetonitn"le R and dilute to 50 rnL with the same
Cream solvent.
PhE" _ Reference solution (a) Dilute 1.0 mL of the test solutionto
100.0 mL with acetonitrile R. Dilute 1.0 mL of this solution
DEFINITION to 10.0 mL with acelOm·trile R.
Content Reference solution (b) Dissolve 30.0 mg of benzoic acidR in
- dibenzoyl peroxide: 70.0 per cent to 77.0 per cent; the mobile phase and dilute to 100.0 mL with the mobile
- water. minimum 20.0 per cent. phase. Dilute 1.0 mL of the solution to 10.0 mL with the
CHARACTERS mobile phase.
Appearance Reference solution (c) Dissolve 50.0 mg of ethylbenzoate R in
White or almostwhite, amorphous or granular powder. the mobile phase and dilute to 100.0 mL with the mobile
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1-286 Benzydamine Hydrochloride 2022
phase. Dilute 1.0 mL of the solution to 100.0 mL with the 1 mL of 0.1 M sodium thiosulfate is equivalent to 12.11 mg of
mobile phase. C I 4HlO04 ·
Reference solutwn (d) Dissolve 50.0 mg of beneoldebyde R in Water (2.5.12)
the mobile phase and dilute to 100.0 mL with the mobile Carry out the semi-micro determination of water, using
phase. Dilute 1.0 mL of the solution to 100.0 mL with the 5.0 rnL of solution (a). Use as the solvent a mixture of
mobile phase. 20.0 mL of anhydrous mechanol Rand 3.0 mL of a 100 gIL
Reference sduuon (e) Dissolve 30.0 mg of benzoic acidRand solution of potassium iodide R in dimechylfannamide R. After
30.0 mg of benzaldehyde R in the mobile phase and dilute to adding solution (a), stir for 5 min before starting the
100.0 mL with the mobile phase. Dilute 1.0 mL of the titration. Carry out a blank determination.
solution to 10.0 mL with the mobile phase. Calculate the percentage content of water using the following
Column: expression:
- size: 1== 0.25 ID, 0 = 4.6 mm;
- statUmary phase: octadecy/silyl silica gelfor chromarcgraphy R (n,-n,)xwx2 + (p x 0.0744)
(10 urn). m
Mobile phase glacial acetic acid R, acetonitrile R, waterR n, number of m~lilitres of iodosulfurous reagent R used in the
sample derermlnation,
(1:500:500 VIVIV). 'Ii me
number of millilitres of iodosulfurous reagent R used in
Flow rate 1 mUmin. blank determination,
w water equivalent of iodosulfurous reagent R in milligrams of
Deuaion Spectrophotometer at 235 nm. water per millilitte of reagent,
InjectUm 20 ~L loop injector. In mass of the substance to be examined used for the preparation
of sojution (a) in grams.
Run time 2 times the retention time of dibenzoyl peroxide. p percentage content of dibenzoyl peroxide.
Relative retention With reference to dihenzoyl peroxide
(retention time == about 28.4 min): impurity B == about 0.15; STORAGE
impurity A = about 0.2; impurity C = about 0.4.
In a container that has been treated to reduce static discharge
System suitability Reference solution (e): and that has a device for release of excess pressure, at a
- resolution: minimum 6 between the peaks due to benzoic temperature of 2 °C to 8 °C J protected from light.
acid and benzaldehyde.
IMPURITIES
Limits:
- impuniy A: not more than the area of the principal peak ('YCHO
(0.25 per cent);
V
- impurity B: not more than the area of the principal peak in A. benzaldehyde,
(1.5 per cent); ('Yco,H
- impurity C: not more man the area of the principal peak
V
(0.25 per cent); B. benzoic acid,
- unspecified impuruies: for each impurity, not more than the
(0.02 per cent).
C. ethyl benzoate.
Chlorides (2.4.4) ___ ~ PIIEIr
Dissolve a quantity of the substance to be examined
containing the equivalent of 0.5 g of dibenzoyl peroxide in
15 mL of acetone R. Add, while stirring, 50 mL of 0.05 M
nitric acid. Allow to stand for 10 min and filter. Wash the
Benzydamine Hydrochloride
residue with 2 quantities, each of 10 ml., of 0.05 M nitric (ph. Bur. monograph 2759)
acid. Combine the filtrate and the washings and dilute to
100 mL With 0.05 M citric acid. Dilute 2.5 mL of the
solution to 15.0 mL with water R.
ASSAY • Hel
Solutian (a) Dissolve 2.500 g immediately before use in
75 mL of dimethylformamide R and dilute to 100.0 mL with
the same solvent. 345.9 /32-69-4
Dibenzoyl peroxide
To 5.0 mL of solution (a) add 20 mL of acetene Rand 3 mL Action and use
of a 500 gIL solution of potassium iodide R and mix. Allow to CycJo-oxygenase inhibitor; analgesic; anti-inflammatory.
stand for 1 min. Titrate with 0.1 M sodium thiosulfate using Preparations
I rnL of starch solution RJ added towards the end of the Benzydamine Cream
titration, as indicator. Carry out a blank titration. Benzydamine Mouthwash
Benzydamine Oromucosal Spray
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2022 Benzydamine Hydrochloride 1-287
PhEII ~ _ Relativeretention With reference to benzydamine (retention

DEFINITION time =: about 12 min): impurity A = about 1.36j
impurity D = about 1.43; impurity B = about 2.0.
3-[(I-Benzyl-IH-indazol-3-yl)oxy]-N,N-dimethylpropan-l-
amine hydrochloride. System suitabl1ilY:
Content
impurities A and D in the chromatogram obtained with
reference solution (b);
PRODUCTION - signal-to-noise ratio: minimum 38 for me principal peak in
Impurity G the chromatogram obtained with reference solution (a).
Maximum 5 ppm, determined by a suitable, validated CaIculau'on of percentage contents:
method. - correction factot»: multiply the peak areas of the following
CHARACTERS impurities by the corresponding correction factor:
Appearance =
impurity A 1.9; impurity D 1.4; =
White or almost white, hygroscopic, crystalline powder. - for each impurity, use the concentration of benzydamine
Solubility
Very soluble in water, freely soluble in ethanol (96 per cent) Limits:
and in methylene chloride, slightly soluble in acetone,
- impun'ty B: maximum 0.5 per cent;
practically insoluble in heptane. - impun'ty A: maximum 0.2 per cent;
- impurity D: maximum 0.15 per cent,
IDENTIFICATION - unsped/ied impurities: for each impurity, maximum
A. Infrared absorption spectrophotometry (2.2.24). 0.10 per cent;
Comparison benzydamine hydrochloride CRS. - total: maximum 1.0 per cent;
B. It gives reaction (a) of chlorides (2.3./). - reporting rhreshold: 0.05 per cent.
TESTS Loss on dryIug (2.2.32)
Related substances
Solvem mixture methanolR, water R (50:50 VIV). Sulfated ash (2.4.14)
examined in the solvent mixture and dilute to 10.0 mL with ASSAY
the solvent mixture. Dissolve 0.300 g in 50 mL of ethanol (96 per <en!! R. Titrate
Reference solution (a) Dilute 1.0 mL of the test solution to with 0.1 M sodium hydroxide, determining the end-point
100.0 mL with the solvent mixture. Dilute 1.0 mL of this potentiometrically (2.2.20).
solution to 10.0 mL with the solvent mixture. I mL of 0./ M sodium hydroxide is equivalent '0 34.59 mg of
Reference sohuion (b) Dissolve 5 mg of benzydamine for C,gH,.CIN,O.
system suitabiNty CRS (containing impurities A, B and D) in STORAGE
5 mL of the solvent mixture. In an airtight container.
Column: IMPURITIES
=
- size: 1= 0.25 m, 0 4.6 mmj
Specified impurities A, B, D, G.
- stationary phase: amidoa/kylsilyl silica gelfor
chromatography R (5 urn). Otherdeuaable impurities (thefollowing substances would, if
Mobile phase:
- mobile phase A: dissolve 1.36 g of potassium dihydrogen
phosphate R in 900 mL of waterfor chromatography R,
monograph Substances for pharmaceuricol use (2034). It is
adjust to pH 3.0 with pltosphoric acidR, add 1.16 g of
sodium oaonesulfonau R and dilute to 1000 mL with water
demonstration of complianu. See also 5.10. Control of impurities
for chromatography R;
in substances for pharmaceutical use) C, E, F.
- mobile phaseB: methanol R;
Tim. Mobile phase A Mobile phase B

(min) (per cent VIII) {per cent Vm
0-2 50 50
2·22 50 ..... 35 50 ..... 65
22 - 29 35 65
A. 3-(dirnethylamino)propyl 2-(benzylamino)benzoate,
Equilibration At least 10 min with the mobile phase at the
initial composition.
Injection 20 ~L.
with benzydamine for system suitability CRS and the
chromatogram obtained with reference solution (b) to B. 3-[(1,5-dibenzyl-lH-indawl-3-yl)oxy]-N,N-
identify the peaks due to impurities A, Band D. dimethylpropan-l-amine,
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1-288 Benzyl Alcohol 2022
.CHARACTERS
Appearance
Clear, colourless, oily liquid.
Solubility
Soluble in water, miscible with ethanol (96 per cent) and
C. l-benzyl-I,2-dihydro-3H-indazol-3-one, with fatty and essential oils.
Relative density
1.043 to 1.049.
IDENTIFICATION
Comparison bellzylalcohol CRS.•
TESTS
tAppearance of solution
D. N'-[3-[(I-benzyl-lH-indazol-3-yl}oxy]propyl)-N',N',N'- Shake 2.0 mL with 60 mL of water R. It dissolves
trimethylpropane-L'I-diamine, completely. The solution is clear (2.2.1) and colourless
(2.2.2, Me/hod /1).•
Acidity
To 10 mL add 10 mL of ,thanol (96 per cent) R and I mL of
phellOlphthal,in solution R. Not more than I mL of 0.1 M
sodium hydroxide is required to change the colour of the
indicator to pink.
Refractive Index (2.2.6)
1.538 to 1.54 I.
Peroxide value (2.5.5, MethodA)
Maximum 5.
E. l-benzyl-2-[3-(dimethylamino}propyl]-I ,2-dihydro-3H-
indezol-S-one, Related substances
Testsolution The substance to be examined.
Standardsolution (a) Dissolve 0.100 g of ,thy/benzell'R in
the test solution and dilute to 10.0 mL with the same
solution. Dilute 2.0 mL of this solution to 20.0 mL with the
test solution.
F. 3-(dimethylamino)propyl 2-aminobenzoate,
Standardsolution (b) Dissolve 2.000 g of di<y<Wh,xy1 R in
the test solution and dilute to 10.0 mL with the same
solution. Dilute 2.0 mL of this solution to 20.0 mL with the
test" solution.
Reference solution (a) Dissolve 0.750 g of beneoldebyde Rand
G.3-cbloro-N,N-dimethylpropan-I-amine. 0.500 g of cydohexylmethanol R in the test solution and dilute
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE" to 25.0 mL with the test solution. Add 1.0 mL of this
solution to a mixture of 2.0 mL of standard solution (a) and
3.0 mL of standard solution (b) and dilute to 20.0 mL with
the test solution.
Benzyl Alcohol 1 Reference solutio" (b) Dissolve 0.250 g of benzaldehyde Rand
0.500 g of cyclohexylmethanol R in the test solution and dilute
(ph. Bur. monograph 0256) to 25.0 mL with the test solution. Add 1.0 mL of this
solutionto a mixture of 2.0 mL of standard solution (a) and
~OH 2.0 mL of standard solution (b) and dilute to 20.0 mL with
V the test solution.
Column:
- moural: fused silica;
G,H,O 108.1 1(j().51-6
- size: I = 30 m, 0 =0.32 nun,
- stationary phase: maaogol 20 000 R (film thickness
Action and use
0.5 pm).
Local anaesthetic; disinfectant.
Canier gas helium for chromatography R.
PIlE" _
Linear velocity 25 cmls at 50 "C.
DEFINITION
Phenylmethanol.
Content
I This morwgraph has undergone pharmacopoeial hannrmiJatj"",

See chapter 5.8. PharmcuopoeiaJ hanncmisation.
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2022 Benzyl Alcohol 1-289
Temperature: -
resohuion: minimum 3.0 between the peaks due to
impurities A and B.
Time Temperature If any peaks in the chromatogram obtained with the test
(min) CC) solution have the same retention times as the peaks due to
Column 0-34 50 ---> 220 ethyl benzene or dicyclohexyl, subtract the areas of any such
34 - 69 220 peaks from the peak areas at these retention times in the
Injection port 200 chromatograms obtained with reference solutions (a) or (b)
Detector 310 (corrected peak areas of ethyl benzene and dicyclohexyl).
Any such peaks in the chromatogram obtained with the test
Detection Flame ionisation. solution are to be included in the assessments for the sum of
Benzyl alcohol not intended for parenteral administration other peaks.
1111'et(ioo Without air-plug, 0.1 JlL of the test solution and Limits:
reference solution (a). - impun·ty A: not more than the difference between the
area of the peak due 10 impurity A in the
Relative retention With reference to benzyl alcohol (retention
=
time ;;;; about 26 min): ethylbenzene about 0.28;
and the area of the peak due to impurity A in the
= =
dicyclohexyl about 0.59; impurity A about 0.68;
chromatogram obtained with the test solution
impurity B = about 0.7 I.
(0.05 per cent);
System suitability Reference solution (a): - impun"ty B: not more than the difference between the
- resolution: minimwn 3.0 between the peaks due to area of the peak due to impurity B in the
impurities A and B. chromatogram obtained with reference solution (b)
If any peaks in the chromatogram obtained with the test and the area of the peak due to impurity B in the
solution have the same retention time as the peaks due to chromatogram obtained with the test solution
ethyl benzene or dicyclohexyl, subtract the areas of any such (0.10 per cent);
peaks from the peak areas at these retention times in the - sum of other peaks with a relative mention less than that
chromatograms obtained with reference solutions (a) or (b) of benzyl alcohol: not more than twice the area of the
(corrected peak areas of ethyl benzene and dicyclohexyl). peak due to ethylbenzene in the chromatogram
Any such peaks in the chromatogram obtained with the test obtained with reference solution (b) corrected if
solution are to be included in the assessments for the sum of necessary as described above (0.02 per cent);
other peaks. - sum of peaks with a relative retention greater than that of
Limits: benzyl alwhol: not more than the area of the peak due
- impun"ey A: not more than the difference between the to dicyclohexyl in the chromatogram obtained with
area of the peak due 10 impurity A in the reference solution (b) corrected if necessary as
chromatogram obtained with reference solution (a) described above (0.2 per cent);
and the area of the peak due to impurity A in the - disregard limir. 0.01 times the area of the peak due to
chromatogram obtained with the test solution ethylbenzene in the chromatogram obtained with
(0.15 per cent); reference solution (b) corrected if necessary as
- impun"ty B: not more than the difference between the described above (0.0001 per cent),
area of the peak due to impurity B in the Residue on evaporation
chromatogram obtained with reference solution (a) Maximum 0.05 per cent.
and the area of the peak due to impurity B in the
After ensuring that the substance to be examined complies
chromatogram obtained with the test solution with the test for peroxide value, evaporate 10.0 g to dryness
(0.10 per cent); in a tared quartz or porcelain crucible or platinum dish on a
- sumof other peaks with a relative retention less than that hot plate at a temperature not exceeding 200°C. Ensure that
of benzyl alcohol: not more than 4 times the area of the the substance to be examined does not boil during
peak due to ethylbenzene in the chromatogram
evaporation. Dry the residue on the hot plate for 1 hand
obtained with reference solution (a) corrected if
allow to cool in a desiccator. The residue weighs a maximum
necessary as described above (0.04 per cent);
of5 mg.
- sum of peaks with a relative retention greater than that of
benzylalcohol: not more than the area of the peak due ASSAY
to dicyclohexyl in the chromatogram obtained with To 0.900 g (m g) add 15.0 mL of a freshly prepared mixture
reference solution (a) corrected if necessary as of 1 volwne of aatit anhydride Rand 7 volumes of anhydrous
described above (0.3 per cent); pyridine R and heat under a reflux condenser on awater-bath
- disregard limit: 0"01 times the area of the peak due to for 30 min. Cool and add 25 mL of water R. Using 0.25 mL
ethylbenzene in the chromatogram obtained with of phenolphthalein solution R as indicator, titrate with I M
reference solution (a) corrected if necessary as sodium hydroxide (n, mL). Carry oul a blank titration
described above (0.0001 per cent). (n2 mL).
Benzyl alcohol intended for parenteral administration Calculate the percentage content of C 7HsO using the
Injection Without air-plug, 0.1 ~L of the test solution and following expression:
reference solution (b)"
1O.81(n, - nil
Relative retention With reference to benzyl alcohol (retention
time = about 26 min): ethylbenzene = about 0.28;
m
dicyclohexyl = about 0.59; impurity A = about 0.68; +STORAGE
impurity B = about 0.7!. In an airtight container, under nitrogen, protected from light
System suitability Reference solution (b): and at a temperature between 2 °C and 8°C"
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1-290 Benzyl Benzoate 2022
LABELLING precipitate is formed that, when washed with water Rand

The label states, where applicable, that the substance is dried in vacuo melts (2.2.1if) at 121 "C to 124 "C.
suitable for use in the manufacture of parenteral C. To the distillate obtained in identification test B add 2.5 g
preparations.• of potassium permangasuue Rand 5 mL of dilute sodium
IMPURITIES hydrox£de solution R. Boil under a reflux condenser for
Specified impuniies A, B. 15 min, cool and filter. Acidify the filtrate with dilute
hydrochloric addR. A white precipitate is formed that, when
~CHO washedwith water R and dried in 'Vacuo, melts (2.2.14) at
121 "C to 124 "C.
V TESTS
Acidity
A. benzaldehyde,
Dissolve 2.0 g in ethanol (96 percent) R and dilute to 10 mL
with the same solvent. Titrate with 0.1 M sodium hydroxide
using phenolphthalel~n sdtuion R as indicator. Not more than
0.2 mL is required to change the colourof the indicator to
pink.
B. cyclohexylmethanol. Relative density (2.2.5)
____________________ 1""" 1.118 to 1.122.
Refractive Index (2.2.6)
1.568 to 1.570.
*****
Freezing point (2.2.18)
Benzyl Benzoate Minimum 17.0 "C.
** **
(ph. Eur. monograph 0705) *** Sulfated ash (2.4.14)
ASSAY
To 2.000 g add 50.0 mL of 0.5 M akaholic potassium
hydroxide and boil gently under a reflux condenser for I h.
Titrate the hot solution with 0.5 M hydrochloric acid using
1 mL of phenolphthalein solution R as indicator. Carry out a
212.2 12()'51-4 blankdetermination.
Action and use I mL of 0.5 M akolwHc potassium hydroxide is equivalent to
Used topically in the treatment of scabies. 106.1 mg of C,.Jf1202.
Preparation STORAGE
Benzyl Benzoate Application In an airtight, well-filled container, protected from light.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
PIlE" _
DEFINITION
Phenylmethyl benzoate.
Content Benzyl Hydroxybenzoate
99.0 per cent to 100.5 per cent. Benzylparaben
CHARACTERS
Appearance o
Colourless or almost colourless crystals or colourless or
almostcolourless, oily liquid.
OHA)
~o~
V
Solubility
Practically insoluble in water, miscible with ethanol
(96 per cent), with methylene chloride and with fatty and
228.3 94-18-8
essential oils.
bp Action and use
About 320 "C. Antimicrobial preservative.
IDENTIFICATION DEFlNfTION
First identification: A. Benzyl Hydroxybenzoace is benzyl 4-hydroxybenzoate.
Second identification: B, C. It contains not less than 99.0% and not more than 101.0%
A. Infrared absorption spectrophotometry (2.2.24). of C14H1203 .
Comparison Ph. Eur. reference spectrum of benzylbenzoate. CHARACTERISTICS
B. To 2 g add 25 mL of akoholic potassium hydroxide A white to creamy white, crystalline powder.
solution R and boil under a refluxcondenser for 2 h. Remove Practically insoluble in water; freely soluble in ethanol (96%)
the ethanolon a water-bath, add 50 mL of water Rand and in ether. It dissolves in solutions of alkali hydroxides.
distill. Collect about 25 mL of distillate and use it for
identification test C. Acidify the liquid remaining in the
distillation flask with dilute hydrochloric acidR. A white
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2022 Benzylpenicillin Potassium 1-291
IDENTIFICATION
A. The infrared absorption spectrum, Appendix II A, is
Benzylpenicillin Potassium
concordant with the reference spectrum of benzyl (ph. Eur. monograph 0113)
hydroxybenzoare (RS 028).
B. The light absorption, Appendix II B, in the range 230 to
350 run of a 0.001 % wlv solution in ethanol (96%) exhibits a
maximum only at 260 om. The absorbance at the maximum
at 260 run is about 0.16.
C. Dissolve 0.1 gin 2 mL of ethanol (96%), boil and add
0.5 mL of nitnc acid solution of mercury. A precipitate is
produced slowly and the supernatant liquid becomes red. 372.5 113-98-4
D. Melongpoinr, about 112°, Appendix V A.
Action and use
TESTS Penicillin antibacterial.
Acidity
Preparadon
Dissolve 0.2 g in 10 mL of ethanol (50%) previously
Benzylpenicillin for Injection
neutralised to methyl red solution and titrate whh 0.1M sodium
hydroxide VS using methyl redsolution as indicator. Not more PhE" _
than 0.1 mL of a.1M sodium hydroxide VS is required to
change the colour of the solution. DEFINlTION
Potassium (2S,5R,6R)-3,3-dimethyl-1-oxo-6-
Related substances
[(phenylacetyl)amino)-4-thia -l-ezabicyclc [3.2.0) heptane-2-
Carry out the method for thin-layer chromawgraphy,
carboxylate.
Appendix ill A, using a plate precoated with silica gel F25 4.1
the surface of which has been modified with chemicaUy- Substance produced by the growth of certain strains of
bonded octadecylsilyl groups (Whatrnan KC 18F plates are Penicillium notatum or related organisms.
suitable) and a mixture of 70 volumes of methanol, Content
30 volumes of water and 1 volume of glacial acetic acid as the 95.0 per cent to 102.0 per cent (dried substance).
mobile phase. Apply separately to the plate 2 pI. of each of CHARACTERS
two solutions of the substance being examined in acetone Appearance
containing (I) 1.0% wlv and (2) 0.010% wlv. After removal
White or almost white,slightly hygroscopic, crystalline
of the plate, allow it to dry in air and examine under powder.
ul"",violet light (254 nm). Any secondary spot in the
chromatogram obtained with solution (1) is not more intense Solubility
than the spot in the chromatogram obtained with Very soluble in water, slightly solublein ethanol
solution (2). (96 per cent), practically insoluble in methylene chloride.
Sulfated ash IDENTIFICATION
Nor more than 0.1 %, Appendix IX A. First identification: A, D.
ASSAY Second idemificauon: B, C, D.
Gently boil 0.12 g under a reflux condenser with 20 mL of A. Infrared absorption spectrophotometry (2.2.24).
2M sodium hydroxide for 30 minutes. Cool and extract with Comparison benzylpenicillin potassium CRS.
wee 20-mL quantities of 1,2-dichloroelhane. Wash the B. Thin-layer chromatography (2.2.27).
combined extracts with 20 mL of O.IM sodium hydroxide and
add the washings to the main aqueous phase, discarding the
examined in 5 mL of waterR.
organic layer. To the aqueous solution add 25 mL of
O.0333M potassium bromate VS,5 mL of a 12.5% w/v solution Reference solution (a) Dissolve 25 mg of benzylpenicillin
of potassium bromide and 10 mL of hydrochloric acid and potassium CRS in 5 mL of water R.
immediately stopper the flask. Shake for 15 minutes and Reference soltuion (1)) Dissolve 25 mg of benzylpenicillin
allow to stand for 15 minutes. Add 25 mL of dilute potassium potassium CRS and 25 mg of phenoxymethylpenicillin
iodide solution and shakevigorously. Titrate the liberated potassium CRS in 5 mL of water R.
iodine with O.IM sodium thiosulfau VS using starch mucilage, PI"", TLC silanised silica gelpIau R.
added towards the end of the titration, as indicator. Repeat Mobile phase Mix 30 volumes of acetone R and 70 volumes
the operation without the substance being examined. of a 154 gIL solutionof ammonium acetate R previously
The difference between the titrations represents the amount adjusted to pH 5.0 with glacial acetic acidR.
of potassium bromate required. The volume of O.0333M
Application I ~L.
potassium bromate VS used is equivalent to half of the volume
of O.lM sodium thiosulfate VS required for the titration. Development Over 2/3 of the plate.
Each mL of O.0333M potassium bromate VS is equivalent to Drying In air.
7.608 mg OfC14H1203' Detection Expose to iodine vapour until the spots appear
System suiuzbility Reference solution (b):
with the test solutionis similar in position, colour and size to
the principal spot in the chromatogram obtainedwith
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1-292 Benzylpenicillin Potassium 2022
C. Place about 2 mg in a test-tube about 150 nun long and Identification of impun'lies Use the chromatogram supplied
15 mm in diameter. Moisten with 0.05 mL of water Rand with benzylpenicillin for'Yrrem suitability CRS and the
add 2 mL of sulfuric acid-fonnaldehyde reagent R. Mix the chromatogram obtained with reference solution (b) to
contents of the tube by swirling; the solution is practically identify the peaks due to impurities A, B, C, D, E, F, G
colourless. Place the test-tube on a water-hath for 1 min; and H.
a reddish-brown colour develops. Relative retention With reference to benzylpenicillin
D. It gives reaction (a) of potassium (2.3.1). (retention time = about 7 min): impurity A = about 0.22;
imputity 0 = about 0.33; impurity C = about 0.35;
TESTS
impurity E = about 0.48 and 0.55; impurity B = about 0.62;
pH (2.2.3)
5.5 to 7.5.
=
impurity F about 0.81 and 0.83; imputity G about 1.47;=
impurity H = about 1.90.
Systemsuitability:
- resolution: minimum 1.2 between me peaks due to me
Appearance of soludon epimers of impurity F and minimum 1.5 between me
The solution is clear (2.2.1). peaks due to impurities D and C in the chromatogram
Dissolve 3.0 g in waterR and dilute to 10 mL with the same obtained with reference solution (b);
solvent. - signal-to-noise ratio: minimum 20 for the principal peak in
Related substances the chromatogram obtained with reference solution (d).
Liquid chromatography (2.2.29), Prepare the solutions Calculation of percentage CQ11tents:
immediately before use. - correaion factors: multiply the peak areas of the following
Test soluTion (a) Dissolve 50.0 mg of the substance to be impurities by the corresponding correction factor:
examined in water R and dilute to 50.0 mL with the same imputity A = 1.4; impurity 0 = 0.6; impurity E = 2.0;
solvent. impurity F 1.7;=
- for each impurity, use the concentration of
benzylpeniciUin potassium in reference solution (c).
solvent. Limits:
- impun"ty F: maximum 2.0 per cent for the sum of the
Reference solution (a) Dissolve 50.0 mg of benzylpenici1lin
2 epimers;
sodium CRS in water R and dilute to 50.0 mL with the same
- ;mpun·ty E: maximum 1.0 per cent for the sum of the
solvent.
isomers;
Reference solution (b) Dissolve 5 mg of benzyipenid1/in for - impun'ty B: maximum 0.5 per cent;
system suirability CRS (containing impurities A, B, C, D, E, - impun'ties A.. C, D, G, H: for each impurity, maximum
F, G and H) in 0.35 mL ofmerhorwlR1 and add 0.65 mL of 0.2 per cent;
water R. - any otherimpuniy: for each impurity, maximum
Reference solution (c) Dilute 1.0 mL of test solution (b) 10 0.2 per cent;
100.0 mL with fOoter R. - wtal: maximum 3.0 per cent;
Reference solution (d) Dilute 1.0 mL of reference solution (c) - reporting threshold: 0.05 per cent.
to 20.0 mL with fOoter R. Loss on drying (2.2.32)
Column: Maximum 1.0 per cent, determined on 1.000 g by drying in
=
- size: / 0.15 m, «2) ;;;; 4.6 mm; an oven at 105°C.
- stationary phase: end-capped octadecylrily/ silica gelfor ASSAY
chromalOgraphy R (3 um),
Mobile phase: Mobile phase Mobile phase A, mobile phase B (70:30 VII').
- mobile phase A: mix 10 volumes of a 68 gIL solution of
potassium d,'hydrogen phosphate R adjusted to pH 3.4 with a Flow rate 1.2 mllmin.
500 gIL solution of phosphoric acidR, 30 volumes of Injection 10 J..lI... of test solution (a) and reference
methanol Rl and 60 volumes of waterfor solution (a).
chromatography R; Calculate the percentage content of C1JI17KN204S taking
- mobile phase B: mix 10 volumes of a 68 gIL solution of into account the assigned content of benzylpenkillin
potassium dihydrogen phosphate R adjusted to pH 3.4 with a sodium CRS and a conversion factor of 1.045.
500 gIL solution of plwsphoric acid R, 35 volumes of fOoter
STORAGE
for chromalOgraphy R and 55 volumes of methanol R1;
In an airtight container. If the substance is sterile.. me
Time Mobile phose A Mobile phase B
(min) (per cent VII') (per eem V/I1 L\1PURITlES
0-7 70 30 Specified impurities A, B, G, D, E.. P, G, H.
7 - 17 70 -t 0 30 ...... 100
17·22 0 100
Flow rate 1.5 mlJmin.

Injection 20!!L of test solution (b) and reference
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2022 Benzylpenicillin Sodium 1-293
A. (2S,5R,6R)-6-amino- 3,3-dimethyl-7-oxo-4-thia-l- H. (2S,5R,6R)-6-(hexanoylamino)-3,3-dimethyl-7-oxo-4-thia-

azabicyclo[3.2.0]heptane-2-carboxylic acid l-azabicyclo[3.2.0]heptane-2-carboxylic acid
(6-aminopeniciUanic acid), (dihydropeniciUin F).
__________ ~ /'liE"
VCO'H
B. phenylacetic acid, Benzylpenicillin Sodium

0't-- ~:/~:,
Dl
~- - H-- S)<CH' o CO-tNa
H-)::"
cn ~· t+s
I H H
N CH3
HO" 0 CH,
~
I 0
H H
C. (ZS,5R,6R)-6-[[(4-hydroxyphenyl)acetyl)amino]-3,3-
dimethyl-7-oxo-4-th ia-l-azabicyclo[3. 2.0]heptane-2-
carboxylic acid, 69-57-8
Action and use
~ XCO,H
Penicillinantibacterial.
Preparadons
>-+8
N'
Ho,C " H
, H
N .><CH3
CH3
BenxylpeniciUin Eye Drops
Benzylpenicillin for Injection
Benzylpenicillin Infusion
D. (3S,7R,7aR)-5-benxyl-2,2-dimethyl-2,3,7,7a- /'liE" ~ _
tetrahydroimidazo[5, I-b] thiazole-3,7-dicarboxylic acid
(penillic acid of henxylpenicillin), DEFINITION
Sodiwn (ZS,5R,6R)-3,3-dimethyl-7-oxo-6-
[(phenylace tyl)amino)-4-thia-I-azabicyclo[3.2.0]heptane-2-
H~.
COzH
~Nt"
H.1. S
CH,
CH3
carboxylate.
Substanceproduced by the growth of certain strains of
o " Co,H
Penicillium notatum or related organisms.
Content
E. (4S)-2-[carboxy[(phenylacetyl)amino)methyl)-5,5-
dimethylthiazolidine-4-carboxyJic acid (penicilloic acids of CHARACTERS
benxylpenicillin), Appearance
White or almost white, hygroscopic, crystalline powder.
H-, Co,H Solubility
HNX--CH3 Verysoluble in water, slightly soluble in ethanol
~ I· '><CH andepimeratC' (96 per cent), practically insoluble in methylene chloride.
. . . . . ·r-s
o "
~
H
3
IDENTIFICATION
First idendfication: A, D.
Second identification: B J C, D.
F. (2RS,4S)-5,5-<1imethyl-2-[[(phenylacetyl)amino]
methyl]thiazolidine-4-carboxylic acid (penilloic acids of A. Infrared absorption spectrophotometry (2.2.24).
benxylpenicillin), Comparison benzylpenicillin sodium CRS.
examined in 5 mL of water R.
Reference solution (a) Dissolve 25 mg of benzylpenicillin
sodium CRS in 5 mL of water R.
Reference solution (b) Dissolve 25 mg of benzylpenicillin
G. (2S,5R,6R)-6-[(3Z)-hex-3-enoylamino]-3,3-dimethyl-7- sodium CRS and 25 mg of phenoxymethylpeniciUin
oxo-4-thia-I-azabicyclo[3.2.0]heptane-2-carboxylic acid) potassium CRS in 5 mL of water R.
Plate TLC sdanised silica gelplateR.
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1-294 Benzylpenicillin Sodium 2022
klobl1e phase Mix 30 volumes of acetone Rand 70 volumes - mobile phase B: mix 10 volumes of a 68 gILsolution of
of a 154 gIL solution of ammonium acetate R previously potassium di!lydrogen phosphate R adjusted to pH 3.4 with a
adjusted to pH 5.0 with glacial acme acid R. 500 gILsolution of phosphoric acid R, 35 volumes of water
Applicacian 1 ~L. for chromatography Rand 55 volumes of methanol Rl,
Deueiopmew Over 2/3 of the plate.
Tim, Mobile phase A MobIle phase B
Drying In air. (min) (per cent VIJ') (per cent VIP)
Detection Expose to iodine vapour until the spots appear 0-7 70 30
and examine in daylight. 7 - 17 70 ..... 0 30 ..... 100
System suirability Reference solution (b): 17 - 22 0 '00
Results The principal spot in the chromatogram obtained Flow rate 1.5 mUmin.
with the test solution is similar in position, colour and size to Deuaion Spectrophotometer at 225 nm.
the principal spot in the chromatogram obtained with Injection 20 JlL of test solution (b) and reference
reference solution (a). solutions (b), (c) and (d).
C. Place about 2 mg in a test-tube about 150 nun long and Identification of impurities Use the chromatogram supplied
15 nun in diameter. Moisten with 0.05 rnL of water Rand with benzylpenicillin for system suitabililY CRS and the
add 2 mL of sulfuric acid-formaldehytk reagent R. Mix the
contentsof the rube by swirling; the solution is practically identify the peaksdue (Q impurities A J B) C) D) E) F) G
colourless. Place the test-tube on a water-bath for 1 min; and H.
a reddish-brown colour develops.
Relativereumion With reference {Q benzylpenicillin
D. It gives reaction (a) of sodium (2.3.1). (retention time == about 7 min): impurity A ;;;; about 0.22;
TESTS = =
impurity D about 0.33; impurity C about 0.35;
pH (2.2.3) = =
impurity E about 0.48 and 0.55; impurity B about 0.62;
5.5 to 7.5. impurity F = about 0.81 and 0.83; impurity G = about 1.47;
impurity H about 1.90.
20 mL with the same solvent. System suitability:
Appearance of solution - resolution: minimum 1.2 between the peaks due to the
The solution is clear (2.2.1). epirners of impurity F and minimum 1.5 between the
peaks due to impurities D and C in the chromatogram
Dissolve 3.0 g in water R and. dilute to 10 mL with the same obtained with reference solution (b);
solvent. - signal-to-noise ratio: minimum 20 for the principal peak in
Related substances the chromatogram obtainedwith reference solution (d).
Liquid chromatography (2.2.29). Prepare the soluciaru Cakukuion of percentage contenlS:
immediately before use. - cotTedion faaon: multiply the peak areas of the following
Test solUlian (a) Dissolve 50.0 mg of the substance to be impurities by the corresponding correction factor:
examined in water R and dilute to 50.0 mL with the same impurity A = 1.3; impurity D = 0.6; impurity E = 2.0;
solvent. impurity F = 1.7;
Testsolution (b) Dissolve 80.0 mg of the substance to be - for each impurity) use the concentration of
examined in water R and dilute to 20.0 mL with the same benzylpenicillin sodium in reference solution (c).
solvent. Limits:
Reference solution (a) Dissolve 50.0 mg of benzylpenicillin - impurity E: maximum 2.0 per cent for the sum of the
sodium CRS in water R and dilute to 50.0 mL with the same isomers;
solvent. - impun'ty F: maximum 1.0 per cent for the sum of the
Reference solution (b) Dissolve 5 mg of benzylpenicillin for 2 epimers;
system suitability CRS (containing impurities A) B) C, D) E) - impun'ty B: maximum 0.5 per cent;
F, G and H) in 0.35 mL of methanol Rl and add 0.65 mL of - impurities A) C, D) G, H: for each impurity) maximum
water R. 0.2 per cent;
- any other ;mpun'ty: for each impurity) maximum
Reference Solulian (c) Dilute 1.0 mL of test solution (b) to
0.2 per cent;
100.0 mL with waler R.
Reference solutian (d) . Dilute 1.0 mL of reference solution (c) - reporting threshold: 0.05 per cent.
to 20.0 mL with lOoter R.
2-Ethylhexanolc acid (2A.2/!)
Column: Maximum 0.5 per cent mlm.
- size: 1 == 0.15 m, 0 == 4.6 mm;
- suuionary phase: end-capped octadecylsi/yl silica gelfor Los. on drying (2.2.32)
ehromotography R (3 urn); Maximum 1.0 per cent, determined on 1.000 g by drying in
- temperature: 50°C. an oven at 105°C.
Mobile phase: ASSAY
- mobile phase A: mix 10 volumes of a 68 giL solution of Liquid chromatography (2.2.29) as described in the test for
potassium d.hydrogen phosphate R adjusted to pH 3.4 with a related substances with the following modifications.
500 giL solution of plwsphoric acidR, 30 volumes of Mobile phase Mobile phase A, mobile phase B (70:30 VW).
methanol Rl and 60 volumes of waterfor Flow rate 1.2 mUmin.
chromalOgraphy R;
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2022 Betacarotene 1-295
Injection 10 ilL of test solution (a) and reference

solution (a).
Calculate the percentage content of CI~I1N2Na04S taking
into account the assigned content of benzylpenicillin
sodium CRS.
STORAGE G. (2S,5R,6R)-6-[(3Z)-hex-3-enoylamino]-3,3-dimethyl-7-
In an airtight container. If the substance is sterile, the oxo-4-thia-I-azabicyclo[3.2.0]heptane-2-carboxylic acid
containeris also sterileand tamper-evident. (iso-penicillin F),
IMPURITIES o ~~H
H'C~~'#s
Spedfied impuruies A, B, C, D, E, F, G, H. N .><CH3
CH,
II H H
o
H. (2S,5R,6R)-6-(hexanoylamino)-3,3-dimethyl-7-oxo-4-thia-
l-azabicyclo[3.2.0]heptane-2-<:arboxylic acid
(dihydsopenicillin F).
A. (2S, 5R,6R)-6-amino-3,3-dimethyl-7 -oxo-d-thie-l-. _ _ _~ _ ~ ~ Pl>E"
(fi-aminopenicillanic acid),
UCo,H Betacarotene ***

*** ***
B. phenylacetic acid,
o H~.
C0:2H
~ ~'#S
N CH3
CH,
I. H H
HO ~ 0 . 536.9 7235-40-7
C. (2S,5R,6R)-6-[[(4-hydroxyphenyl)acetyl]amino]-3,3- Action and use

dimethyl-7-oxo-4-thia-I-azabicyclo[3.2.0] heptane-2- Precursor of vitamin A.
carboxylic acid, PhE" _
DEFINITION
Main component 1,1 '-[(IE,3E,5E,7E,9E,IIE,13E,15E,17E)-
3,7,12, 16-tetramethyloctadeca-l ,3,5,7,9,11,13,15,17-
nonaene-I,I8-diyl]bis(2,6,6-trimeiliylcyclohexene) (~,~
carotene, betacarotene, al/-trans-betacarotene, all-E-
beracarotene).
Content
D, (3S, 7 R, 7aR)-5-benxyl-2,2-dimethyl-2,3,7,7a- 95.0 per cent to 102.0 per cent (dried substance).
tetrahydroimidazo[5,I-b]thiazole-3,7-dicarboxylic acid
(peniDic acid of benxylpenicillin), CHARACTERS
Appearance
H. Co,H Brown-red or brownish-red, crystalline powder.
HN~CH3 Solubility
~~rs CH, Practically insoluble in water, soluble in tetrahydrofuran,
V ;; Co,H
practically insoluble in anhydrous ethanol.
It is sensitive to air, heatand light, especially in solution.
E. (4S)-2-[carboxy[(phenylacetyl)amino]methyl)-5,5- IDENTIFICATION
dimethylthiazolidine-4-carboxylic acid (penicilloic acids of Carry ou' all operations as rapidlY as possible. Prepare the
benzylpenicillin), solutions immediately be/ore use.
H-, C0:2H (2.2.25).
HW~Y-
..x ~ I·
GH
3
andepimeralC'
Test solution Dissolve 50.0 mg in terrahydrofuran Rand
(jr'l( -""'fS 3 CH dilute to 100.0 mL with the same solvent. Dilute 10.0 mL of
V ;; the solution to 50.0 mL with tetmhydrofuran R. Dilute
3.0 mL of this solution to 100.0 mL with cydohexane R.
F. (2RS,4S)-5,5-<limethyl-2-[[(phenylacetyl)amino]
Absorption maximum 455 om.
methyl]thiazolidine-4-carboxylic acid (peniUoic acids of Shonlder About 427 nm.
benzylpenicillin), Absorbance ratio A4sslA483 = 1.14 to 1.18.
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1-296 Betacarotene 2022
B. Examine the chromatograms obtained in the assay. Limits:

Results The principal peak in the chromatogram obtained - impun·ty E: maximum 4.0 percent;
with the test solution is similar in retention time and size to - impurity D: maximum 2.0 per cent;
the principal peak in the chromatogram obtained with - impun·ty G: maximum 0.6 per cent;
reference solution (a). - impurity G: maximum 0.4 per cent;
- impurity H: maximum 0.4 per cent;
Cony au' all operations as rapidlY as poss.ble. Prepare the 0.15 per cent;
solutions immediately before use. - sum of impurities other than D and E: maximum
Related substances 5.0 per cent;
Liquid chromatography (2.2.29): use the normalisation - reporting threshold: 0.05 per cent (0.5 times the area of the
procedure. principal peak in the chromatogram obtained with
Stabilised te.rahydrofuran tetrahydrofuran R containing reference solution (c».
0.25 gIL of buty/hydroxytoluene R. The thresholds indicated underRelated substances
Test solutkm Dissolve 50.0rog of the substance to be (Table 2034.-1) in the general monograph Substances for
examined in stabilised tetrahydrofuran and dilute to phannaceutical use (2034) do not apply.
200.0 mL with the same solvent. Dilute 5.0 mL of the Loss on drying (2.2.32)
solution to 100.0 mL with anhydrous ethanol R. Maximum 0.2 per cent, determined on 1.000 g by drying in
Reference solution (a) Dissolve 50.0 mg of hewarolene CRS vacuo at 40 ·C for 4 h.
in stabilised tetrahydrofuran and dilute to 200.0 mL with the Sulfated ash (2.4.14)
same solvent. Dilute 5.0 mL of the solution to 100.0 mL Maximwn 0.1 per cent, determined on 2.0 g.
with anhydrous ethanolR.
ASSAY
Reference solution (b) Dissolve 5 mg of beuuaYOtene for system
Liquid chromatography (2.2.29) as desctibed in the test for
sm·t4bility CRS (containing impurities C, D, E, G and H) in
related substances with the following modification:
stabilised tetrahydrofuran and dilute to 20 mL with the same
solvent. Dilute 1 mL of the solution to 20 rnL with anhydrous Injection Test solution and reference solution (a).
ethanol R. Calculate the percentage content of CtOH56 taking into
Reference solution (c) Dilute 1.0 mL of the test solution to account the assigned content of beuuarotene CRS.
100.0 mL with stabilised tetrahydrofuran. Dilute 1.0 mL of STORAGE
this solution to 10.0 mL with anhydrous ethanol R. In an airtight container, protected from light.
Column: IMPURITIES
- size: 1= 0.25 m, 0 = 4.6 mm; Specified impurities C, D) E, G, H.
- suuionaty phase: amidoalkylsilYl silica gelfor
chromatography R (5 um); Other detectable impuri.., (the following substances would, if
- temperature: 30 ·C. present at a sufficient level, be deteaedby one or other of the tests
Mobile phase Dissolve 50 mg of burylhydroxytoluene R in
criterion for other/unspecljied impuniies. It is therefore no'
20 mL of 2-propanol R, then add 0.2 mL of N,N-
necessary to identify these impun·ties for demonstration of
diisopropylethylamine R. 25 mL of a 2.0 gIL solution of
compliance. See also5.10. Conlrol of ;mpun'ties in substances for
ammonium acetate R, 455 mL of atetonitrik Rand 450 mL of
phannaceutical use) A, B, F.
methanol R; warm to room temperature and dilute to
1000 mL with methanolR. CH, CH,
FkJw rate 0.6 mIlmin.
Detection Spectrophotometer at 448 ron.
CH,
Injection 20 ilL of the test solution and reference CH,
Run time 2.7 times the retention time of betacarotene. A. (2E,4£,6E,8E,IOE,12E)-2,7,II-trimethyl-13-(2.6,6-
Identification of impu~;ties Use the chromatogram supplied trime thylcyclohex-I-en-I-yl)trideca-2,4,6,8, I0, 12-hexaenal
with betaearolene for system suitabilil)l CRS and the (12' -apo-Bcaroten-Iz' -all,
identify the peaks due to impurities C, D, E, G and H.
Relative retention With reference to betacarotene (retention
time :::: about 28 min): impurity G = about 0.6;
impurity C = about 0.95; impurity D = about 1.05;
impurity E = about 1.15; impurity H = about 2.4.
- resolution: minimum 1.5 between the peaks due to B. 1-[(1£,3E,5E,7E,9E,IIE,13E,15E,17E,19E)-
impurity C and betacarotene; 3,7,12, 16,20,24-hexamethylpentacosa-
- peak-to-Valley ratio: minimum 1.5, where H p = height 1,3,5,7,9,11,13,15,17,19,23-undecaen-l-yl]-2,6,6-
above the baseline of the peakdue to impurity D and trimethylcyclohexene (P,'¥-carotene, y-caro[ene),
H!1 = height above the baseline of the lowestpoint of the
curve separating this peak from the peakdue to
betacarotene.
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2022 Betadex 1-297
Betadex
Betacyclodextrin
C. 1,3,3-trimethyl-2-[(lE,3E,5E, 7E,9E,liE, l3E, 15E,17Ej-
3,7, 12, I 6-telramethyl-1 8-[ (IR)-2,6,6-trimethylcyclohex-2-
en-l-yl]octadeca-l J3,S,7 ,9,11,13,15,17-nonaen-l-yl]
cyclohexene «6'R)-P,£-carotene, a-carotene),
D. 1,1 '-[(lE,3E,5E,7E,9E,l1E,l3E,15Z,17Ej-3,7,12,16-
tetramethyloctadeca-l,3,5,7,9,11,13,15, 17-nonaene-I,18- 7585-39-9
1135
diyl]bis(2,6,6-trimethylcyclohexene) (9-ci<-p,ll-carotene,
9-cU-betacarotene), Action and use
Carner moleculefor drugdelivery systems.
CH, CH,
"'[>1 _
DEFINITION
Cycloheptakis-(l ....4)-(o<-D-g1 ucopyranosyl)
(cyclomaltoheptaose or p-cyclodextrin).
Content
98.0 per cent to 102.0 per cent (dried substance),
CHARACTERS
Appearance
White or almostwhite, amorphous or crystalline, hygroscopic
powder.
E. 1,1 '-[(lE,3E,5E,7E,9E,IIZ,13E,15E,17Ej-3,7,12,16-
tetramethyloetadeca-I,3,5,7,9,11,13,15, 17-nonaene-l,18- Solubility
diyl]bis(2,6,6-trimethylcyclohexene) (13-eis-p,p-carotene, Sparingly soluble in water and in propylene glycol, practically
13-cis-betacarotene), insoJuble in anhydrous ethanol and in methylene chloride.
IDENTIFICATION
H~
CH,
B. Examine the chromatograms obtained in the assay.
with test solution (b) is similar in retention time and size to
the principaJ peak in the chromatogram obtained with
C. Dissolve 0.2 g in 2 mL of iodine solution R4 by warming
on a water-bath, and allow to stand at room temperature.
H,C A yellowish-brown precipitate is formed.
H,C
TESTS
Solution S
Dissolve 1.000 g in carbon dioxide-free waterR with heating,
F. 1,I'-[(IE,3E,5E, 7E,9Z,llE,13E,15E,17Ej-3,7,12,16- allow to cool and dilute to 100.0 mL with the same solvent.
tetramethyloetadeca-I,3,5,7,9,11,13,15,17 -nonaene-l,18-
diyl]bis(2,6,6-trimethylcyclohexene) (15-cis-p,p-carotene,
Solution S is clear (2.2.1).
15-cis-betacarotene),
pH (2.2.3)
G. unknown structure (oxidation products),
5.0 to 8.0.
H. unknown structure.
To 10 mL of solution S add 0.1 mL of a saturated solution
_ _ _ _ _--'- Ph[>I
of potassium chloride R.
Speclflc optical rotation (2.2.7)
+ 160 to + 164 (dried substance), determined on solutionS.
Reducing sugars
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1-298 Betadex 2022
Testsolution To 1 mL of solution S add 1 rnL of cupn"- - sum of impurities other thanA and B: maximum
tartaric solution R4. Heat on a water-bath for 10 min, cool to 0.5 per cent;
room temperature. Add 10 mL of ammonium molybdate - reporting threshold: 0.15 per cent.
reagent Rl and allow to stand for 15 min. Residual solvents
Reference solution Prepare a reference solution at the same Head-space gas chromatography (2.2.28): use the standard
time and in the same manner as the test solution, using additions method.
I mL of a 0.02 gIL solution of gluco" R. Internal standard ethylene chloride R.
Measure the absorbance (2.2.25) of the test solution and the Stock solUlion A To 20 ~L of ethylene chloride R add 0.5 mL
reference solution at the absorption maximum at 740 nm of dimethyl sulfoxide R and dilute to 25.0 mL with waterR.
using water R as the compensation liquid. The absorbance of
Stock solution B To 25 I1L of trichloroethylene R add 25 ~L
the test solution is not greater than that of the reference
of toluene Rand 0.5 mL of dimethyl sulfoxide R, then dilute 10
solution. 50.0 mL with waterR.
Light-absorbing impurities Test solutions (a), (b), (c) and (d) In each of 4 identical
Examine solution S between 230 run and 750 om. Between vials, introduce 0.5 g of the substance to be examined, 0.10 g
230 run and 350 run, the absorbance (2.2.25) is not greater of calcium chloride R, 30 J.lL of a.-amylase solution Rand 1 mL
than 0.10. Between 350 run and 750 nm, the absorbance of reference solutions (a), (b), (c) and (d), respectively, then
(2.2.25) is not greater than 0.05. add 9.0 mL of waterR. Prepare test solutions (b), (c) and (d)
Related substances in triplicate.
Liquid chromatography (2.2.29). Reference solution (a) Dilute 250 ~L of stock solution A to
Test solutUm (a) Dissolve 0.250 g of me substance to be 10.0 mL with water R.
examined in water R with heating, cool and dilute to Reference solution (b) To 100 ~L of Slack solution B add
25.0 mL with the same solvent. 250 ~L of stock solution A and dilute to 10.0 mL with
Test solution (b) Dilute 5.0 mL of test solution (a) to waterR.
50.0 mL with waterR. Referenu solution (c) To 200 I1L of stock solution B add
Reference solution (a) Dissolve 25.0 mg of alfadex CRS 250 ~L of stock solution A and dilute to 10.0 mL with
(impurity A). 25.0 mg of gommacydodexuin CRS water R.
(impurity B) and 50.0 mg of betadex CRS in waterR, then Reference solution (d) To 300 ~L of stock solution B add
dilute to 50.0 mL with the same solvent. 250 ~L of stock solution A and dilute to 10.0 mL with
Reference solution (b) Dilute 5.0 mL of reference solution (a) water R.
to 100.0 mL with waterR. . Blank so/utum In a vial identical to those used for the test
Reference solution (c) Dissolve 25.0 mg of betadex CRS in solutions, introduce 0.10 g of calcium chloride R, 30 I1L of
water R and dilute to 25.0 mL with the same solvent. «-amylase solution R, 0.5 mL of dimethyl sulfoxide Rand
Column: 10.0 mL of waterR.
- size: I =0.25 m, 0 = 4.6 mm; Column:
- stationary phase: end-capped octadecylsilyl silica gelfor - material: fused silica;
chromatography R (10 urn). - size: 1= 25 m, 0 = 0.32 mm:
Mobile phase methanol R, waterfor chromatography R - stationary phase: maaogol 20 000 R (film thickness I urn).
(10:90 VIV). Carrier gas helium for chromatography R.
Flow rate 1.5 mUmin. Flowrate I. 7 mllmin.
Detection Differential refractometer. Static head-space conditions that may be used: if theequipment
Equilibration With the mobile phase for about 3 h. has different setting parameters, adjust the equipment seuings so as
Injection 50 tIL of test solution (a) and reference to comply with the system suitability ctitetion:
solutions (a) and (b). - equilibration temperature: 45°C;
- equilibration time: 2 h;
Run time 1.5 times the retention time of betadex.
- syringe temperature: 50°C;
Identification of impuriues Use the chromatogram obtained - injeuion speed: 500 ~Us.
with reference solution (a) to identify the peaksdue to
Temperature:
impurities A and B.
- column: 50°C;
Relative retention With reference to betadex (retention - injutUm port: 140°C;
= =
time about 10 min): impurity B about 0.3; - detector. 280 'C.
Injection 200 ~L.
impurities B and A; if necessary, adjust the concentration Relativeretention With reference to the internal standard
of methanol in the mobile phase. (retention time = about 13 min): trichloroethylene = about
0.6; toluene = about 0.8.
- for impurities A and B, use the concentration of the System suitability Test solutions (b), (c) and (d):
corresponding impurity in reference solution (b); - repeatability: maximum relative standard deviations of the
- for impurities other than A and B, use the concentration ratios of the areas of the peaks due to trichloroethylene
of betadex in reference solution (b). and toluene to that of the peak due to ethylene chloride
of 10.0 per cent, for each set of triplicate test solutions
Limits:
and each residual solvent.
- impwlUes A, B: for each impurity, maximum
0.25 per cent;
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2022 Betahistine Dihydrochloride 1-299
Calculate the content of trichloroethylene and of toluene

Betahistine Dihydrochloride ***
taking their relative densities to be 1.46 and 0.87, ** **
respectively. (Ph. Eur. monograph 1665) *****
Limits:
- trichkJroethyiene: maximum 10 ppm;
- toluene: maximum 10 ppm.
V
r"'N~~, CH3.2HCI

an oven at 120 "C for 2 h. 209.1 5579-84-0
Maximum 0.1 per cent, determined on 1.0 g. Action and use
Histamine HI receptor antagonist; antihistamine.
ASSAY
Preparation
Liquid chromatography (2.2.29) as described in the lest for
Betahistine Dihydrochloride Tablets
Injuu"on Test solution (b) and reference solutions (a) PhEII _
and (c).
DEFINITION
System suitability Reference solution (a): N-Methyl-2-(pyridin-2-yl)ethanamine dihydsochloride.
- rtpMrability: maximum relative standard deviation of
2.0 per cent for the area of the peak due to betadex, Content
determined on 5 injections.
Calculate the percentage content of [C 6H100sh using the CHARACTERS
chromatogram obtained with reference solution (c) and Appearance
taking into account the assigned content of betadex CRS. White or slightly yellow powder, very hygroscopic.
STORAGE Solubility
In an airtight container. Very soluble in water, soluble in ethanol (96 per cent),
practically insoluble in 2-propanol.
IMPURITIES
Specijied impurities A, B. IDENTIFICATION
Second identijieatWn: A, C, D.
OOOOr-<0q:Q':"
'pHOH B. Infrared absorption spectrophotometry (2.2.24).
Comparison betahistine dihydrochloride CRS.
\--no
0ri0H HO 0
OH . C. Thin-layer chromatography (2.2.27).
~'o HO OH 0 .
0 "':Q~OO HO OH
examined in 2 mL of ethanol (96 per em!! R.
Reference solution Dissolve 10 mg of betahistine
dihydrochkJride CRS in 2 mL of ethanol (96 per em!! R.
Pia,. TLC ,,7ica gel GFm pia,. R.
Mobile phase concentrated ammonia R, ethyl acetate R,
methanol R (0.75:15:30 VIVII').
A. cyclohexakis-(I ~4)-("-D-glucopyranosyl) (alfadex or
cyclomalrohexaose or c-cyclodextrin), Applicatwn 2 IlL.
Drying At 110 "C for 10 min.
reference solution.
TESTS
Solution S
Dissolve 5.0 g in carbon dioxide-free water R, and dilute to
than reference solution Ba (2.2.2, MethodII).
B. cyclooctakis-(l ~4)-("-D--glucopyranosyl) pH (2.2.3)
(cyclomaltooctaose or y-cyclodextrin). 2.0 to 3.0 for solution S.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'I>EII Related substances
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1-300 Betahistine Mesilate 2022
Test solution Dissolve 25 mg of the substance to be IMPURITIES

examined in the mobile phase and dilute to 25.0 mL with Specified impurities A, B, C.
the mobilephase.
Reference solution (aJ Dissolve 10 mg of betahistine
dihydrochloride CRS and 10 mg of 2-vinylpyridine R in the
(JCH 2
mobile phase and dilute to 50.0 mL with the mobile phase.

Dilute 2.0 mL of the solution to 50.0 mL with the mobile A. 2-ethenylpyridine (2-vinylpyridine),
phase.
Reference solutifm (b) Dilute 1.0 mL of the test solution to
Reference solution (c) Dilute 2.0 mL of refecence solution (b)
to 10.0 mL with the mobile phase. B. 2-(pyridin-2-yl)ethanol,
Column:
- size: 1::::; 0.15 m, 0 ;:: 3.0 mrn,
- stationary phase: base-deactivated end-capped octadecy/silyl
silica gelfor chromatography R (5 urn).
Mobile phase Dissolve 2.0 g of sodium dodecyl sulfate R in a
mixture of 15 mL of a 10 per cent VIV solution of su/fum C. N-methyl-2-(pyridin-2-yl)-N-[2-(pyridin-2-yl)ethyl)
acid R, 35 mL of a 17 gIL solution of tetrabutylammonium ethanamine.
hydrogen sulfate Rand 650 mL of waterR; adjust to pH 3.3 _~ P1>E"
using dilute sodium hydroxide solution R and mix with 300 mL
of cuelOnitrik R.
Flow rate I mUmin.
***
**• *••
Detection Spectrophotometer at 260 nm. Betahistine Mesilate
Injection 20~.
* ••
Run time 4 times the retention time of betahistine.
Relative retention With reference to betahistine (retention
N ~
= =
impurity A about 0.3; impurity C about 3.
V'CH,.
- resolution: minimum 3.5 between the peaks due to 328.4 54856-21-4
2-vinylpyridine and betahistine.
Limits: Action and use
- cotreaion factor. for the calculation of content. multiply the Histamine HI receptor antagonist; antihistamine.
peak area of impurity B by 0.4; P1>E" _
- impurilies A, B, C: for each impurity, not more than the
area of the principal peak in the chromatogram obtained DEFINTI10N
with reference solution (c) (0.2 per cent); N-Methyl-2-(pyridin-2-yl)ethanamine bis(methanesulfonate).
- unspecified impurities: for each impurity, not more than Content
0.5 times of the acea of the psincipal peak in the 98.0 per cent to 101.0 per cent (anhydrous substance).
(0.10 per cent); PRODUCTION
- total: not more than0.5 times the area of the principal It is considered that alkyl methanesulfonate esters are
peak in the chromatogram obtained with reference genotoxicand are potential impurities in betahistine mesilate.
solution (b) (0.5 per cent); The manufacruring process should be developed taking into
- disregard limir: 0.25 times the area of the principal peak in consideration the principles of quality risk management,
the chromatogram obtained with reference solution (c) together with considerations of the quality of starting
(0.05 per cent). materials, process capability and validation. The general
methods 2.5.17. Methyl, ethyl and isopropyl methan<7U!ionaii in .
methanesulfonic acid, 2.5.18. Methyl, ethyland isopropyl
methanesulfonate in activesubstances and 2.$.39.
an oven at 105°C.
l\tIethanesulfonyl chloride in methanesulfomc iicid areavailable to
Sulfated ash (2.4.14) assist manufacturers.
CHARACTERS
ASSAY Appearance
Dissolve 80.0 mg in 50 mL of ethanol (96 per cen!! R. Titrate White or almostwhile, crystalline powder, very hygroscopic.
with 0.1 M sodium hydroxide, determining the end-point
Solubility
potentiometrically (2.2.20). Read the volume added to reach
Verysoluble in water, freely solublein ethanol (96 per cent),
the second point of inflexion.
very slightly soluble in 2-propanol.
CsH14C12N2' IDENTIFICATION
STORAGE
Second idenufication: A, C, D.
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2022 Betahistine Mesilate 1-301
B. Infrared absorption spectrophotometry (2.2.24). Injection 20 ~L.

Comparison betahistine mesikue CRS. Run time 3 times the retention time of betahistine mesilate.
C. Thin-layer chromatography (2.2.27). Retention time Betahistine mesilate ;;;; about 8 min.
Test solution Dissolve 10 mg of the substance to be System suitability Reference solution (a):
examined in ethanol (96 per <en!> R and dilute to 2 mL with - resolution: minlmum 3.5 between the peaks due to
the same solvent. impurity A and betahistine mesilate.
Reference solution Dissolve 10 mg of betahistine me.siJate CRS Limits:
in ethanol (96 percen!> R and dilute to 2 mL with the same - impurity A: not more than the area of the principal peak
solvent. in the chromatogram obtainedwith reference solution (c)
Plate TLC silica gelFZ54 plate R. (0.2 per cent);
Mobile phase concentrated ammonia R, ethylacetate R,
- unspecified impun"tie.s: for each impurity, not more than
methanol R (0.75:15:30 VIVIV).
chromatogram obtainedwith reference solution (b)
Applicalion 2 ~L. (0.10 pet cent);
Development Over 3/4 of the plate. - total: not more than 0.5 times the area of the principal
Drying At 110 °C for 10 min. peakin the chromatogram obtained with reference
Detection Examine in ultraviolet light at 254 om. solution (b) (0.5 pet cent);
(0.05 per cent).
principal spot in the chromatogram obtained wirh the
reference solution. 2-Propanol (2.4.24)
D. To 0.1 g add 5 mL of dilute ftydrochloric acidR and shake Maximum 0.5 per cent.
for about 5 min. Add 1 mL of barium chlaride sdution R 1. Chlorides (2.4.4)
The solution remains clear. To a funher 0.1 g add 0.5 g of Maximum 35 ppm.
anhydrous sodium carbonate R, mix and igniteuntil a white To 14 mLofsolution S add 1 mLofwaterR.
residue is obtained. Allow to cool and dissolve the residue in
Sulfates (2.4. 13)
7 mL of water R. The solution gives reaction (a) of sulfates
Maximum 250 ppm.
(2.3.1).
Dilute 6 mLofsolutionS to 15 mL with distiUedwaterR.
TESTS
Water (2.5.12)
Solution S Maximum 2.0 per cent, determined on 0.50 g.
Dissolve 5.0 g in carbon dioxide-free water R prepared from
distilled water R, and dilute to 50 mL with the same solvent. ASSAY
Appearance of solution Dissolve 0.140 gin 50 mL of a mixtwe of I volume of
Solution S is clear (2.2.1) and colourless (2.2.2, Methad If). anhydrous acetic acid Rand 7 volumes of acetic anhydride R.
Titratewith 0.1 M perchlork acid, determining the end-point
pH (2.2.3) potentiometrically (2.2.2U).
Related substances of C,oH,oN,O.Sz.
STORAGE
Test soluticn Dissolve 50 mg of the substance to he
the mobile phase. IMPURITIES
Reference solution (a) Dissolve 10 mg of betahistine Specified impurities A.
mesiiaeCRS and 10 mg of 2.."inylpyridine R (impurity A) in
phase. Dilute 2.0 mL of this solution to 50.0 mL with the ~CH'
mobile phase.
A. 2-ethenylpyridine (2-vinylpyridine).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "'E<r
Column:
- size: 1= 0.25 m, 0 = 4.6 rom;
- stationary phase: ocuule<y/si/yl silica gelfor chromatography R
(5 um).
Mobile phase Dissolve 2.0 g of sadium dadecyl sulfate R in a
mixture of 15 volumes ofa 10 per cent VIVsolution of
sulfuric acidR, 35 volumes of a 17 gIL solution of
tetrabutylammonium ftydrogen sulfate Rand 650 volumes of
water R; adjust to pH 3.3 using dilute sadium hydroxide
solution R and mix with 300 volumes of aceumitrile R.
Flow rale I mUmin.
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1-302 Betamethasone 2022

Betamethasone with the test solutionis similar in position, colourand size to
(ph. Eur. monograph 0312) the principal spot in the chromatogram obtained with the
reference solution.
C. Add about 2 mg to 2 mL of sulfuric acid R and shake to
dissolve. Within 5 min) a deep reddish-brown colour
develops. Add this solution to 10 mL of water R and mix.
The colour is discharged and a clear solution remains.
TESTS
o Specific optical rotation (2.2.7)
+ liB to + 126 (dried substance).
392.5 378-44-9 Dissolve 0.125 gin memanol R and dilute to 25.0 mL with
the same solvent.
Action and use
Glucocorticoid. Related substances
Preparation
Test sdudon Dissolve 25.0 mg of the substance to be
Betamethasone Tablets
examined in a mixture of equalvolumes of aceumitrile Rand
PIlE" _ methanol R and dilute to 10.0 mL with the same mixture of
solvents.
DEFINITION
9-Fluoro-ll p,17,21-trihydroxy-16p-methylpregna-I,4-diene- Reference so/urion (a) Dissolve 2 mg of betamethasone CRS
and 2 mg of methylprednisolone CRS in mobile phase A and
3,2o-dione.
dilute to 100 rnL with mobile phase A.
Content
Reference so/urion (b) Dilute 1.0 rnL of the test solution to
100.0 rnL with mobile phase A.
CHARACTERS Column:
Appearance - size: I = 0.25 m, 0 = 4.6 mm,
White or almostwhite, crystalline powder. - stationary phase: end-capped oetade<y/silyl silica gelfor
Solubility chnnnatQgraphy R (5 urn);
PracticaUy insoluble in water} sparingly soluble in anhydrous - temperature: 45 "C.
ethanol, very slightly soluble in methylene chloride. Mobile phase:
It shows polymorphism (5.9). - mobile phaseA: mix 250 rnL of acetQnitn7e Rand 700 mL
of waterfor chromawgraphy R and allow to equilibrate;
IDENfIFICATION
dilute to 1000 rnL with waterfor chnnnatQgraphy Rand
First itkn.Jjcalwn:A, B. mix;
Second itkmijicarion: B, C. - mobik phase B: aceumitrile R;
A. Infrared absorption spectrophotometry (2.2.2<f).
Comparison betamethasone CRS. Time MobUe phase A MobUe phase B
(min) (per cent JlIJI) (per cent JlIV)
0-15 100 0
15 - 40 100 -+ 0 0-+ 100
substance separately in the minimum volumeof methylene
40 - 41 0-+ 100 100 ..... 0
chlotid» R, evaporate to dryness on a water-bath and record
41- 46 100 0
Flow rale 2.5 mUmin.
examined in the mobile phase and dilute to 10.0 mL with Detection Spectrophotometer at 254 run.
the mobile phase. EquilibratiJJn With mobile phase B for at least 30 min and
Reference solution Dissolve 10 mg of betamethasone CRS in then with mobile phase A for 5 min; for subsequent
the mobile phase and dilute to 10.0 mL with the mobile chromatograms, use the conditions described from 40 min to
phase. 46 min.
Plate TLC silica gel F m plate R. Injection 20 J.I1.; inject a mixture of equal volumes of
acetQnitrik R and methanol R as a blank.
Mobile phase methanol R, methylene chloride R (10:90 VIV).
Retention time Methylprednisolone = about 11.5 min;
Applicarion 5 ~L. betamethascne = about 12.5 min.
Drying In air. - resolution: minimum 1.5 between the peaks due to
Deteaion Spray with a solution prepared as follows: dissolve methylprednisolone and betamethasone; if necessary)
0.25 g of 2,4-dihydroxybenzaldehyde R in glacial acetic acidR, adjust the concentration of acetonitrile in mobilephase A.
dilute to 50 mL with the same solvent and add a mixture of Limits:
12.5 mL of sulfUric acid Rand 37.5 mL of glacial acetic - impun"ties A, B~ C, D~ E, F, G, H, I~ J: for each impurity)
add R; heat at 90 DC for 35 min or until the spots appear, not more than the area of the principal peakin the
allow to cool and examine in daylight and in ultraviolet light chromatogram obtained with reference solution (b)
at 365 nm. (1.0 per cent), and not more than I such peakhas an
area greater than 0.5 times the area of the principal peak
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2022 Betamethasone 1-303

(0.5 per cent);
- total: not more than twice me area of the principal peak in
(2.0 per cent);
- disregard limit: 0.05 times the area of the principal peak in o
(0.05 per cent). E. 9,11 p-epoxy-17,21-dihydroxy-16p-methyl-9p-pregoa-1 ,4-
Loss on drying (2.2.32) diene-3,ZQ-dione,
an oven at 105°C.
ASSAY
Dissolve 0.100 g in ethanol (96 percent) R and dilute to
solution to 100.0 mL with ethanol (96 per cent) R. Measure o
the absorbance (2.2.25) at the absorption maximwn at
238.5 om. F. 17,21-dihydroxy-16I3-methylpregoa-1 ,4, II-triene-3,20-
Calculate the content of C22H29FOs taking the specific dione,
absorbance [0 be 395.
o
STORAGE
IMPURITIES
Sp«ified impun"lies A, 8.1 C, D, E, F, 0, H, I, J.
o
G. II~, 17,21-trihydroxy-16p-methylpregoa-I,4-diene-3,20-
dlone,
A. 9-fluoro-11 p,17,21-trihydroxy-I6«-methylpregoa-I,4-
diene-3,2Q-dione (dexamethasone),
o
H. l4-fluoro-ll p, 17,21-trihydroxy-16p-methyl-8~,9P, 1413-
pregna-I A-diene-3,20-dione,
B. zf-chloro-c-Buoro-j I p, 17-dihydroxy-Ifijl-methylpregna-
l,4-diene-3,20-dione,
o
I. 8-fluoro-11 p,17,21-trihydroxy-16p-methyl-8~,9p-pregoa
1,4-dieJ;le-3,20-dione,
o
o
C. 17,21-dihydroxy-16p-methylpregoa-I,4,9(I I)-triene-3,20-
dione,
o
J. 17,21-dihydroxy-16I3-methylpregoa-I,4-diene-3,20-dione.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEU'
o
D. 9-fluoro-11 p,17-dihydroxy-16p-methyl-3,20-dioxopregoa-
1,4-dien-21-yl ethoxycarboxylate,
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1-304 Betamethasone Acetate 2022
C. Add about 2 mg to 2 mL of sulfwic acid R and shake to

Betamethasone Acetate dissolve. Within 5 min, a deep reddish-brown colour
(Ph. Eur. monograph 0975) develops. Add this solution to 10 mL of water R and mix.
The colouris discharged and a clear solutionremains.
TESTS
Specific optical rotation (2.2. 7)
+ 120 to + 128 (anhydrous substance).
Dissolve 0.250 g in diaxon R and dilute to 25.0 mL with the
same solvent.
o
Related substances
434.5 987-24-6
Action and use examined in 4 mL of acetonitrile R and dilute to 10.0 mL
Glucocorticoid. with the same solvent.
P>E" _ Reference solution (a) Dissolve 2 mg of betamethasone
acetate CRS and 2 mg of dexamethasone acetate CRS
DEFINITION (impurity B) in the mobile phase and dilute to 100 mL with
9-Fluoro-11 p,l7-dihydroxy-16Il-methyl-3,2o-dioxopregna- me mobile phase.
l,4-dien-21-yl acetate. Reference solution (b) Dilute 1.0 mL of the test solution to
Content 100.0 mL with the mobile phase.
97.0 per cent to 103.0 per cent (anhydrous substance). Column:
CHARACTERS - size: I;;;: 0.25 m, 0 ;;;: 4.6 mm;
Appearance - stationary phase: end-capped oaad«ylsilylsilica gelfor
White or almost white, crystalline powder. chromatography R (5 pm).
Solubillty
Mobile phase Mix 380 mL of auwnitrile Rand 550 mL of
waterfor chromatography R and allow to equilibrate; dilute to
1000 mL with waterfor chroma",graphy R and mix.
soluble in ethanol (96 per cent) and in methylene chloride.
Flow rate 1 mUmin.
IDENfIFICATION
Injection 20 l'1..
Run time 2.5 times the retention time of betamethasone
Second idemification: B, C.
acetate.
Retention time Betamethasone acetate e about 19 min;
Comparison betamethasone acetate CRS. impurity B = about 22 min.
If the spectra obtained in the solid state show differences, System suitalrility Reference solution (a):
dissolve me substance to be examined and the reference - resolution: minimum 3.3 between the peaks due to
substance separately in the minimum volume of methanol R, betamethasone acetateand impurity B,; if necessary, adjust
evaporate to dryness on a water-bath and record new spectra slightly the concentration of acetonitrile in the mobile
using the residues. phase.
B. 'Thin-layer chromatography (2.2.27). Limits:
TestsolutUm Dissolve 10 mg of the substance to he - impun·ties A, BJ CJ D: for each impurity, not more than
examined in the mobile phase and dilute to 10.0 mL with 0.5 times the area of the principal peak in the
the mobilephase. chromatogram obtained with reference solution (b)
Reference solution Dissolve 10 mg of betomethasone (0.5 per cent);
a"'uue CRS in the mobile phase and dilute to 10.0 mL with - total: not more than 1.25 times the area of the principal
the mobile phase. peak in the chromatogram obtainedwith reference
Plate TLC silica gelF254 plate R. solution (b) (1.25 per cent);
Mobile phase methanol R, methylene chloride R (10:90 VIV).
Application 5 l'1.. (0.05 per cent).
Development Over 3/4 of the plate. Water (2.5.12)
Drying In air. Maximum 4.0 per cent, determined on 0.100 g.
Deteaion Spray with a solution prepared as follows: dissolve ASSAY
0.25 g of 2,4-dihydroxybenza/dehyde R in glacial acetic acidR,
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to
dilute to 50 mL with the same solvent and add a mixture of 100.0 mL with the same solvent. Dilute 2.0 mL of the
12.5 mL of sulfuric acid Rand 37.5 mL of glacial acetic
solution to 100.0 mL with ethanol (96 per cent) R. Measure
acid R; heat at 90 °C for 35 min or until the spots appear,
the absorbance (2.2.25) at the absorption maximum at
allow to cool and examinein daylight and in ultraviolet light
240 nm.
at 365 nm.
Calculate the content of C24H31F06 taking the specific
Results The principal spot in the chromatogram obtained absorbance to be 350.
the principal spot in the chromatogram obtained with the STORAGE
reference solution. Protected from light.
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2022 Betamethasone Dipropionate 1-305
IMPURITIES PhE" ~
Specified impurities A, B, C, D. DEFINITION

o 9-Fluoro-11 ~-hydroxy-16Il-methyl-3,20-dioxopregna-I,4
diene-17,21-diyl dipropanoate.
Content
CHARACTERS
Appearance
Whiteor almost white, crystalline powder.
A. 9-f1uoro-II~, 17,21-trihydroxy-16~-methylpregna-I,4
diene-3,2Q-dione (betarnethasone), Solublllty
Practically insoluble in water, freely soluble in acetone and in
methylene chloride, sparingly soluble in ethanol
(96 per cent).
IDENTIFICATION
o Second identification: B, C.
B. 9-f1uoro-II~, 17-dihydroxy-I6ct-methyl-3,2o-dioxopregna- Comparison besamethasone dipropionate CRS.
l,4-dien-21-yl acetate (dexamethasone acetate),
the mobilephase.
Reference solution Dissolve 10 mg of betamethasone
dipropionale CRS in the mobile phase and dilute to 10.0 mL
o with the mobile phase.
Plate TLC silica gelFZ54 plate R.
C. 9-f1uoro-17-hydroxy-Itill-merhyl-3,20-dioxopregna-I,4-
diene-l ljkz l-diyl diacetate (betamethasone 11,21- Mobile phase methanol R, methylene chloride R (10:90 VIV).
diacetate), Application 5!1L
Deuelopmem Over 3/4 of the plate.
o 0
Drying In air.
CH3 0-'<.
/'-.'---\-·OH CH3 Detection Spray with a solution prepared as follows: dissolve
', CH3 0.25 g of 2,4-dihydroxybmza/dehyde R in glacial acetic acidR,
""" H diluteto 50 mL with thesame solvent and add a mixture of
o 12.5 mL of sulfuric acidR and 37.5 mL of glacial acetic
acid Rj heat the plateat 90 °C for 35 min or until the spots
D. 9,llll-epoxy-17-hydroxy-16~-methyl-3,20-dioxo-9~ appear, allow to cool and examine in daylight and in
pregna-l,4-d.ien-21-yl acetate. ultraviolet light at 365 run.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" Results The principal spot in the chromatogram obtained
reference solution.
Betamethasone Dipropionate C. Add about 2 mg to 2 mL of ru!furic acid R and shake to
dissolve. Within 5 min, a deep reddish-brown colour
(Ph. Eur. monograph 0809) develops. Add this solution to 10 mL of water R and mix.
The colour is discharged and a clear solution remains.
TESTS
Dissolve 0.250 g in anhydrous ethanol R and dilute to
Related substances
o liquid chromatography (2.2.29).
5593-20-4 Test solution (a) Dissolve 60.0 mg of the substance to be
C,.H"FO, 504.6
Action and use the mobile phase.
Glucocorticoid. Test solution (b) Dilute 1.0 mL oftest solution (a) to
Preparations 10.0 mL with the mobile phase.
Calcipotriol and Betamethasone Cutaneous Foam Reference solution (a) Dissolve 5 mg of betamethasone
Calcipotriol and Betamethasone Gel dipropionate for system suitability A CRS (containing
Calcipotriol and Betamethasone Ointment
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1-306 Betamethasone Dipropionate 2022
impurities B, C, D, E, G and I) in the mobile phase and - impurities D, E, G: for each impurity, not more than twice
dilute to 2 mL with the mobile phase. the area of the principal peak in the chromatogram
Reference solution (b) Dilute 1.0 mL of test solution (a) to obtained with reference solution (b) (0.2 per cent);
100.0 mL with the mobile phase. Dilute 1.0 mL of this - impurity I: not more than 1.5 times the area of the
solution to 10.0 mL with the mobilephase. principal peak in the chromatogram obtained with
Reference solution (c) Dissolve 60.0 mg of beiomabosone
diprr>Pionate CRS in the mobile phase and dilute to 25.0 mL - unspecified impurities: for each impurity, not more than the
with the mobile phase. Dilute 1.0 mL of the solution to area of the principal peak in the chromatogram obtained
Reference solution (d) Dissolve 5 mg of betamethasone peak in the chromatogram obtained with reference
dipropionate for peak idenlification CRS (containing solution (b) (1.0 per cent);
impurity H) in the mobile phase and dilute to 2 mL with the - disregard limit 0.5 times the area of me principal peak in
mobile phase.
me chromatogram obtained with reference solution (b)
Column: (0.05 per cent).
- size: 1= 0.10 m, 0 = 2.0 mm;
- slationary phase: end-capped octadecylsilyl silica gelfor
Maximwn 1"0 per cent, determined on 0.500 g by drying in
chromatography R (2.5 11m);
an oven at 105°C.
- temperature: 20 ± 2°C.
Mobile phase Mix 35 mL of water for chromatography Rand ASSAY
56 mL of aceumitn"le R and allow to equilibrate; dilute to Liquid chromatography (2.2.29) as described in the test for
100 mL with waterfor chromatography R and mix. related substances with me following modification.
Flow rate 0.2 mUmin. Injection Test solution (b) and reference solution (c).
Detection Spectrophotometer at 254 nm. Calculate the percentage content of C2sH37F01 taking into
account the assigned content of beunnethasone
Injection 5 JlL of test solution (a) and reference
dipropionate CRS.
solutions (a), (b) and (d).
Run time 3 times the retention time of betamethasone STORAGE
dipropionate. Protected from light.
Identification of impurities Use the chromatogram supplied IMPURITIES
with betamethasone diprapionate for system ",ilabih''Y A CRS Specified impurities B, C, D, E, G, H, I.
and the chromatogram obtained with reference solution (a) Other deC<Clabie impun;ies (rhe following subslances would, if
to identify the peaks due to impurities B, C, D, E, G and I; present at a sufficient level, be detected by oneor other of the leSts
use the chromatogram supplied with betamethasone in the monograph. They are limited by thegeneral acapumu
dipropionate for peak identification CRS and the chromatogram criterion for o/herlunspe<ified impurities ondlor by thegeneral
obtained with reference solution (d) to identify the peakdue monograph SubslancesforpharmaceuticaJ use (2034).1, is
to impurity H. therefore not neussary to identify these impun'ties for
Relative retention With reference to betamethasone demonstration of camp/innce. See also 5.10. Control of impurities
dipropionate (retention time = about 10 min): in subscances for pharmaceutical use) A, F.
impurity D = about 0.7; impurity I = about 1.16;
impurity E = about 1.22; impurity H = about 1.7j
=
impurity G about 2. I.
- peak-/Q-VaJley ratio: minimum 2.0, where H. = height
above the baselineof the peak due to impurity I and
HI} = height above the baselineof the lowest point of the o
betamethasone dipropionate; minimum 4.0, where A. 9-l1uoro-Il~, 17,21-trihydroxy-16Il-methylpregna-I,4-
Hp = height above the baselineof the peak due to diene-3,20-dione (betamerhasone),
impurity I and H; = height above the baselineof the
lowest point of the curve separating this peak from the
peak due to impurity E.
Limits:
- correction factors: for the calculation of content, multiply
corresponding correction factor: impurity G = l.3j o
impurity H = 1.4;
- impun'ty C: not more than 5 times the area of the B. 9-l1uoro-l1 ~,21-dihydroxy-16~-methyl-3,20-dioxopregna
principal peak in the chromatogram obtained with 1,4-dien-17~yl propanoate (betamethasone 17-propionate),
- impurities B, H: for each impurity, not more than 3 times
obtained with reference solution' (b) (0.3 per cent);
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2022 Betamethasone Sodium Phosphate 1-307
o
o o O~CH3
~,.".C,H_3-< ... O~CH3
. CH 3
H
o o
H 'Br
C. 9-ftuoro-II~, 17-<lihydroxy-16~-methyl-3,20-dioxopregna
1,4-d.ien-21-yl propanoate (betamethasone 21-propionate), H. 6~-bromo-9-ftuoro-II ~-hydroxy-16~-methyl-3,20-
dioxopregna-l,4-diene-11,21-diyl dipropanoate (00-
bromobetarnethasone dipropionate),
o
o °o~CH3
..._O~CH3
\ CH3
/ ' d ,,...,....',"-""'/ H
o
o
D. 9-ftuoro-11 Jl-hydroxy-16Jl-methyl-3,20-dioxopregna-I,4-
dlene-I?,21-diyl zl-acetate 17-propanoate 1. c-fiuoro-t I ~-hydroxy-16~-methyl- 3,20-dioxopregn-4-ene-
(betamethasone zl-acetate 17-propionate), 17,21-diyl dipropanoate (l,2-dihydrobetamethasone
dipropionate).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Betamethasone Sodium
Phosphate
o (Ph. Bur. monograph 0810)
E. 9-eWoro-llJl-hydroxy-16~-methyl-3,20-dioxopregna-I,4
diene-17,21-diyl dipropanoate (beclometasone
dipropionate),
516.4 151-73-5
Action and use

Glucocorticoid.
Preparations
F. 9,11 ~-epoxy-16~-methyl-3,2o-dioxo-9~-pregna-I,4-diene Betamethasone Eye Drops
17,21-diyl dipropanoate (9~,11~-epoxybetamethasone Betamethasone Injection
dipropionate), Betamethasone Sodium Phosphate Tablets
PIlE" _
DEFINlTION
Disodium 9-ftuoro-II~,11-dihydroxy-16~-methyl-3,20
dioxopregna-I,4-dien-21-yl phosphate.
Content
96,0 per cent to 103.0 per cent (anhydrous substance).
o CHARACTERS
Appearance
G. 9-ftuoro-16Jl-methyl-3,20-dioxopregna-1,4-<1iene- White or almost white, veryhygroscopic powder.
11p,17,2I-triyl tripropanoate (betamethasone
SolubUlty
tripropionate),
Freely soluble in water, slightly soluble in ethanol
(96 per cent), praeticaUy insoluble in methylene chloride.
IDENTIFICATION
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1-308 Betamethasone Sodium Phosphate 2022
A. Infrared absorption spectrophotometry (2.2.24). Reference solution (b) Dilute 1.0 mL of the test solution to
Comparison betamethasone sodium phosphate CRS. 50.0 mL with the mobile phase.
If the spectra obtained in the solid state show differences, Column:
dissolve the substance to be examined and the reference - size: 1= 0.25 m, 0 = 4.6 mm;
substance separately in me minimum volume of ethanol - stationary phase: end-capped octad«y/siryl silica gelfor
(96 per cenV R, evaporate to dryness on a water-hath and chromatography R (5 pm).
record new spectra using the residues. MoMe phase In a 250 mL conical flask, weigh 1.360 g of
B. Thin-layer chromatography (2.2.27). potassium dihydrogen phosphate Rand 0.600 g of hexy/amine R,
mix and allow lO stand for 10 min and then dissolve in
185 mL of water for chromatography R; add 65.mL of
examined in methanol R and dilute (0 10.0 mL with the same
acetonitrile R, mix and filter (0.45 urn).
solvent.
Flow rate I mUmin.
Reference solution Dissolve 10 mg of beuuneduuone sodium
phosphate CRS in methanol R and dilute to 10.0 mL with the Detection Spectrophotometer at 254 nD1.
same solvent. Equilibration With the mobile phase for about 45 min.
Plate TLC silica gel F m plate R. Injection 20 ur,
Mob,le phase glacial acetic acid R, water R, butanol R Run time Twice the retention time of betamethasone
(20:20:60 VIV/V). sodium phosphate.
Applicalion 5 ~L. Retention time Betamethasone sodiumphosphate = about
Development Over 3/4 of the plate. 14 min; dexamethasone sodium phosphate = about
15.5 min.
Dlying In air.
Detection Spray with a solution prepared as foUows: dissolve System suitability Reference solution (a):
0.25 g of 2,4-dihydroxybenzaldehyde R in glacial a",ic acidR,
betamethasone sodiumphosphate and dexamethasone
dilute to 50 mL with the same solvent and add a mixture of
sodium phosphate; if necessary, increase the concentration
12.5 mL of suJjuri< acidRand 37.5 mL of glacial acenc
of acetonitrile R or increase the concentration of waterfor
acid R; heat the plate at 90 °C for 35 min or until the spots
appear and allow to cool; examine in daylight and in chromatography R in the mobile phase.
ultraviolet light at 365 nm. Limits:
Results The principal spot in the chromatogram obtained - any impun'ty: for each impurity, not more than the area of
reference solution (b) (2 per cent), and not more than
1 such peakhas an area greater than 0.5 times the area of
reference solution.
C. Add about 2 mg to 2 mL of su/juri< acidR and shake to reference solution (b) (1 per cent);
dissolve. \Vithin 5 min, an intense reddish-brown colour - total: not more than 1.5 times the area of the principal
develops. Add the solution to 10 mL of waterR and mix. peak in the chromatogram obtained with reference
The colourdisappears and a clearsolution remains. solution (b) (3 per cent);
TESTS - disregard limir. 0.025 times the area of the principal peak
Solution S in the chromatogram obtained with reference solution (b)
Dissolve 1.0 g in carbon dioxide-free waterR and dilute to (0.05 per cent).
20 mL with the same solvent. Inorganic phosphate
Appearance of solution Maximum 1 per cent.
Solution S is clear (2.2.1) and not more intensely coloured Dissolve 50 mg in water R and dilute to 100 mL with the
than referencesolution B 7 (2.2.2, Method 1l). same solvent. To 10 mL of the solutionadd 5 mL of
pH (2.2.3) molybdovanadic reagent R, mix and allow to stand for 5 min.
7.5 to 9.0. Any yellow colour in the solutionis not more intense than
that in a standard prepared at the same time and in the same
Dilute I mL of solution S to 5 mL with carbon dioxide-free
manner using 10 mL of phosphate standard solunon (5 ppm
waterR.
PO,) R.
Water (2.5.12)
Dissolve0.250 g in waterR and dilute to 25.0 mL with the
same solvent. ASSAY
Dissolve 0.100 g in water R aod dilute to 100.0 mL with the
Related substances
same solvent. Dilute 5.0 mL of the solution to 250.0 mL
with water R. Measure the absorbance (2.2.25) at the
Test solution Dissolve 62.5 mg of the substance to be absorption maximum at 241 om.
Calculate the content of C22H28FNa20sP taking the specific
the mobile phase.
absorbance to be 297.
Reference solution (a) Dissolve 25 mg of betamethasone
sodium phosphate CRS and 25 mg of dexomethasone sodium STORAGE
phosphate CRS in the mobile phase and dilute to 25 mL with In an airtight container, protected from light.
the mobile phase. Dilute I mL of this solution to 25 mL _ _ _ _ _ _- - - - - - - - - - - - - - PIlE"
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2022 Betamethasone Valerate 1-309
Betamethasone Valerate *** Solvent mixture glacial acetic acid R, mobile phase
*** *** (1:1000 VW).
(Ph. Eur. monograph 0811) *** Test solution Dissolve 50 mg of the substance to be
Reference solution (a) Dilute 1.0 mL of the 'est solution [0
solutionto 10.0 mL with the solventmixture.
Reference solution (b) Dissolve 12.5 mg of betamethasone
o valerate for system suitability CRS (containing impurities D
and G) in 5.0 mL of the solvent mixture. Use 1.0 mL of this
solution to dissolve the contents of a vial of betamethasone
476.6 2152-44-5
valerate impun"ty mixture CRS (containing impurities C, H
Action and use and\).
Glucocorticoid. Reference solution (c) Dissolve 6 mg of betamethasone CRS
(impurity A) and 3 mg of betamethasone 21-iJalerate CRS
Preparations
(impurity E) in 30.0 mL of the solvent mixture. Dilute
Betarnethasone and Clioquinol Cream
1.0 mL of this solution to 10.0 mL with the solvent mixture.
Betamethasone and Clioquinol Ointment
Column:
Betamethasone Valerate and Coal Tar Paste - size: I =0.25 m, 0 =4.6 mm;
Betamethasone Valerate Cream ~ stationary phase: end-eapped octade<y/silyl silica gelfor
Betamethasone Valerate Lotion chromalOgraphy R (5 1JOl);
Betamethasone Valerate Ointment - temperature: 20 "C.
Betamethasone Valerate Scalp Application Mobile phase aceumitriJe R, water R (50:50 VIII).
F/gw rate I mIlrnin.
PhE<I _
DEFINITION Inj«tion 20 ~L.
9-Fluoro-11 p,21-dihydroxy-16J}-methyl-3,20-dioxopregna- Run time 2.5 times the retention time of betamethasone
1,4-dien-17-yl pentanoate. valerate.
Content Idetluficat«m of impurities Use the chromatogram supplied
97.0 per cent '0 103.0 per cent (dried substance). with betamethasone valerate for system suitability CRS and the
CHARACTERS chromatogram obtained with reference solution (b) to
Appearance identify the peaksdue to impurities C, D, GJ H and I; use
White or almost white, crystalline powder. the chromatogram obtained with reference solution (c) to
identify the peaks due to impurities A and E.
Solubility
Practically insoluble in water, freely soluble in acetone and in Relative retention With reference to betamethasone valerate
methylene chloride, soluble in ethanol (96 per cent). (retention time =about 20 min): impurity A = about 0.3;
impurity I = about 0.6; impurity C = about 0.8;
mp
impurity H = about 1.3; impurity D = about 1.4;
About 192 °C, with decomposition.
impurity E = about 1.6; impurity G = about 2.0.
IDENTIFICATION System suitability Reference solution (b):
A.lnfrared absorption spectrophotometry (2.2.24). - resolution: minimum 1.7 between the peaksdue to
Comparison betamethasone 17-iJa/erate CRS. impurities Hand D.
If me spectra obtained in the solid state show differences, Limits:
dissolve me substance to be examined and the reference - impurity A: not more than 7 times the area of the
substance separately in the minimum volume of methylene principal peak in the chromatogram obtained with
chloride R, evaporate to dryness on a water-bath and record reference solution (a) (0.7 per cent);
new spectra using the residues. - impurities E G: for each impurity, not more than 3 times
J
B. Examine the chromatograms obtained in the test for the area of the principal peak in the chromatogram
relatedsubstances. obtained with reference solution (a) (0.3 per cent);
- impurities C, H, I: for each impurity, not more than
chromatogram obtainedwith reference solution (a)
(0.15 per cent);
reference solution (b).
TESTS area of the principal peak in the chromatogram obtained
Specific optical rotation (2.2.7) with reference solution (a) (0.10 per cent);
+ 77 to + 83 (dtied substance). - total: not more than 15 times the area of the principal
Dissolve 0.250 g in anhydrous ethanol R and dilute to peak in the chromatogram obtained with reference
25.0 mL with the same solvent. solution (a) (1.5 per cent);
Related substances - disregard limit: 0.5 times the area of the principal peak in
Liquid chromatography (2.2.29). Cany ou' the test protected
(0.05 per cent).
from ligh< Prepa.. the solutions immediately before use.
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1-310 Betamethasone Valerate 2022

o
an oven at 105°C.
O~CH3
ASSAY
Dissolve 50.0 109 in ethanol (96 per «nO R and dilute to
solution to 50.0 mL with ethanol (96 per «nO R. Measure the o
absorbance (2.2.25) at the absorption maximum at 240 nm.
Calculate the contentof C27H37F06 taking the specific E. c-fluorc-l Ijl, 17-dihydroxy-16~-methyl-3,2o-dioxopregna
absorbance to be 325. 1,4-dien-21-yl pentanoate (beramethasone 21-valerate),
STORAGE
IMPURITIES
Specified impurities A, C, E, OJ H, 1.
Otherdetectable impuniies (thefollawing substances wenld, if
present at a sufficient level, be deuaed by oneor other of the tests
o
in themonograph. They are limited by thegeneral acceptance
cntetion for other/unspecified impurities and/or by thegeneral F. 21-hydroxy-16~-methyl-3,2o-dioxopregna-I,4,9(l1 j-trien-
1~-yl pentanoate (betamethasone valerate a-9(1l»,
therefore not necessary to Ukmify these impurilies for
demonstration of compliance. See also 5. I O. Control of impun'ties
in substances for pharmaceutical use) B, D, F.
o
G. 6,,-bromo-9-fluoro-II~,21-dihydroxy-16~methyl-3,2o
dioxopregna-l,4-dien-17-yl pentanoate (6ct-bromo-
A. 9-fluoro-ll~, l7,21-trihydroxy-16~-methylpregna-I,4
betamethasone valerate),
diene-3.,20-dione (betamethasone),
OH
o
.",.~'L"".... O~CH3
\ cH,
'-'/1"--"'--/ H
o o
B. 9-fluoro-ll~, 17-dihydroxy-16~-methylpregna-1 ,4-diene- H. 9-<:Woro-11 ~,21-dihydroxy-16~methyl-3,20-dioxopregna
3,20-dione (21-deoxy-betamethasone), l,4-dien-17-yl pentanoate (beclomethasone 17-valerate),
OH OH
o o 0
~CH, CI-I:I ~CH3
'0 ····0
\ H
0.., /"'- II/''---/ CH,
o o
C. 9-fluoro-ll ~,21-dihydroxy-16,,-methyl-3,20-dioxopregna I. 9-fluoro-11 ~,21-dibydroxy-3,20-dioxopregna-I,4-dien-17
1,4-dien-17-yl pentanoate (dexamethasone 17-valerate), yl pentanoate (9-fluoro-prednisolone 17-valerate).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII
OH
o
.... O~CH3
\ CH3
H
D. 9-bromo-11 ~,21-dihydroxy-16~methyl-3,2o-dioxopregna
1,4-dien-17-yl pentanoate (9-bromo-betamethasone
valerate),
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2022 Betaxolol Hydrochloride 1-311
Results B The principal spot in the chromatogram obtained

Betaxolol Hydrochloride with me test solution is similar in position, colour and size to
(Ph. Bur. monograph 1072) me principal spot in the chromatogram obtained with
HCI
TESTS
MethodIf).
343.9 63659-19-8 Dissolve 0.5 g in waterR and dilute to 25 mL with the same
solvent.
Action and use Acidity or alkalinity
Beta-adrenoceptor antagonist. Dissolve 0.20 g in carbon dioxide-free water R and dilute to
Preparations 20 mL with the same solvent. Add 0.2 mL of methyl red
Betaxolol Eye Drops, Solution solution Rand 0.2 mL of 0.01 M hydro<hlcri< acid.
Betaxolol Eye Drops, Suspension The solution is red. Add 0.4 mL of 0.01 M sodium hydroxide.
The .solution is yellow.
?hE,; _
Related substances
DEFINITION Liquid chromatography (2.2.29). Prepare reference solutions (c)
(2RS)-I-[4-[2-( Cyclopropylmethoxy)ethYl/phenoxy]- 3-[( 1- and (d) immediately before use.
methylethyl)amino]propan-2-ol hydrochloride. Test solution Dissolve 10 mg of the substance to be
Content examined in the mobilephase and dilute to 5.0 mL with the
98.5 per cent to 101.5 per cent (dried substance). mobilephase.
Reference solution (a) Dissolve 8 mg of the substance to be
CHARACTERS
examined and 4 mg of betaxolol impurity A CRS in 20.0 mL
Appearance
of the mobile phase.
Solublllty 100.0 mL with the mobile phase.
Very soluble in water, freely soluble in ethanol (96 per cent),
Reference solution (c) Dissolve 2 mg of betaxo101
soluble in methylene chloride.
impurity C CRS in 50 mL of the mobile phase. Dilute 5 mL
IDENTIFICATION of the solutionto 20 mL with the mobilephase.
First identification: B, D. Reference solution (d) Dissolve 10 mg of becaxololfor peak
Second identification: A, C D.
J identification CRS (containing impurities B, D and E) in
A. Melting point (2.2.14): 113 ·C to 117 ·C. 5 mL of reference solution (c).
B. Infrared absorption spectrophotometry (2.2.24). Column:
Comparison becaxolol hydro<hloride CRS. - size: 1= 0.25 m, 0 = 4 mm;
- stationary phase: octy/silyl silica gelfor chromawgraphy R
C. Thin-layer chromatography (2.2.27). (5 urn),
Test solution Dissolve 10 mg of the substance to be Mob,7e phase Mix 175 mL of autonitrile Rand 175 mL of
examined in 1 mL of methanol R. methanol R and dilute to I L with a 3.4 gIL solution of
Reference solution (a) Dissolve 20 mg of betaXoloi potassium dihydrogen phosphate R, previously adjusted to
hydrochlcride CRS in 2 mL of methanol R. pH 3.0 with phosphori< acidR.
Reference solution (b) Dissolve 10 eng of oxprenolc1 Flow rate 1~5 rnUmin.
hydrochloride CRS in I mL of reference solution (a). Detection Spectrophotometer at 273 run.
Plcte TLC octade<ylsilyl silica gelFm plate R. Injection 20 ul., of the test solutionand reference
Mobile phase perchlom acidR, methanol R, water R solutions (a), (b) and (d).
(0.5:50:50 VIV/V). Run time 4.5 the retention time of betaxolol,
Application 2 ~L. Idemification of impuruies Use the chromatogram obtained
Development Overa path of 10 em. withreference solution (a) to identify the peak due to
Drying In air. impurity A; use the chromatogram supplied with betaxolol for
System suitability Reference solution (b): peak identification CRS and the chromatogram obtained with
- the chromatogram shows 2 clearly separated spots. reference solution (d) to identify the peaks due to
impurities B, C, D and E.
Detection A Examine in ultraviolet light at 254 run.
Results A The principal spot in the chromatogram obtained Relative retention Withreference to betaxolol (retention
time : : :; about 8 min): impurity B = about 0.3;
impurity A = about 0.8; impurity D = about 1.5;
solution (a).
= =
impurity E about 2.2; impurity C about 4.1.
Detection B Spray with a 50 gIL solution of vanillin R in a
mixture of 5 volumes of sulfuric acidR, 10 volumes of glacial
impurity A and betaxolo!.
acetic acid Rand 85 volwnes of methanol R, heat at
100-105 °C until the colour of the spots reaches maximum
intensity (l0-15 min), and examine in daylight.
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1-312 Bezafibrate 2022
Limits:
- impuriues A, B, C, D, E: for each impurity, not more than
(0.3 per cent);
- unspecified impun"oo: for each impurity J not more than E. (2RS)-I-[ 4-(2-butoxyethyl)phenoxy]-3- [(1-
methylethyljaminojpropan-z-ol.
chromatogram obtained with reference solution (b) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII
(0.10 per cent);
- touzI: not more than the area of the principal peak in the
(1.0 per cent};
- disregard limit: 0.05 times the area of the principal peak in Bezafibrate
(0.05 per cent). (ph. Bur. monograph 1394)
Maximum 0.5 per cent, detennined on 1.000 g by drying in
an oven at 105°C.
ASSAY
361.8 41859-67-0
hydrochloric acid and 50 mL of elhanol (96 per cen!! R. Carry Action and use
our a potentiometric titration (2.2.20). using 0.1 M sodium Fibrate; lipid-regulating drug.
Preparations
inflexion.
Bezafibrate Tablets
Bezafibrate Prolonged-release Tablets
of C,sH,oCIN03 .
PIIEII _
STORAGE
Protected from light. DEFINITION
IMPURITIES 2-[4-[2-[(4-CWorobenzoyl)amino]ethyl]phenoxy]-2-
Specified impun"ties AJ BJ CJ D, E. methylpropanolc acid.
Content
H OH 98.0 per cent to 102.0 per cent (dried substance).
~
O~ ~
1? I Y CH, andenantiomer CHARACTERS
~c CH] Appearance
A. (2RS)-I-(4-ethylphenoxy)-3-[(I-methylethyl) Solubility
a~o]pDOpan-2-0~ Practically insoluble in water, freely soluble in
dlmethylformamide, sparingly soluble in acetone and in
H pH H ethanol (96 per cent). It dissolves in dilute solutions of alkali
~
O~ N
?'" I Y CH, anden:nliomer hydroxides.
HO C~ It shows polymorphism (5.9).
IDENTIFICATION
B. (2RS)-I-[4-(2-hydroxyethyl)phenoxy]-3-[(1- First identification: AJ B.
methylethyl)amino]propan-2-01, Second identification: A, C.
A. Melting point (2.2.14): 181 'C to 185 'C.
Comparison bezofibraie CRS.
If the spectra obtainedshow differences, dissolve the
C. (2RS)-2-[[4-[2-(cyclopropylmethoxy)ethyl] separately in methanol R and evaporate to dryness. Dry the
phenoxy]methyl]oxirane, residues in vacuo at 80°C for 1 h and record new spectra
using the residues"
examined in methanol R and dilute to 5_ mL with the same
solvent.
Reference solutWn Dissolve 10 mg of bezafibrate CRS in
D.4-[2-(cyclopropylmethoxy)ethyl]phenol,
methanol R and dilute to 5 mL,~~ lI,le, s~e, solvent,
Pia", TLC silica gel Fm pia", R._ ..
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2022 Bezafibrate 1-313
Mobile phase glacial acetic acidR, methylethyl ketone R, - total: not more than 1.5 times the area of the principal
xylene R (2.7:30:60 VIVIV). peak in the chromatogram obtained with reference
Application 5 ~L. solution (a) (0.75 per cent);
Development Over half of the plate. - disregard limit: 0.1 times the area of the principal peakin
Drying At 120 "C for at least 15 min. (0.05 per cent).
Chlorides (2.4.4)
Resulu The principal spot in the chromatogram obtained Maximum 300 ppm.
with the test solutionis similar in position and size to the
Dilute 10 mL of solution S to 50 mL with water R. Filter the
resultant suspension through a wet filter previously washed
reference solution.
with waterR until free from chlorides. Prepare the standard
TESTS using 9 mL of chloride standard solution (5 ppm Cl) R and
Solution S 6 mL of water R.
Dissolve 1.0 g in dimethylfonnamide R and dilute to 20 mL Loss on drying (2.2.32)
with the same solvent. Maximum 0.5 per cent, determined on 1.000 g by drying in
Appearance of solution an oven at 105°C.
Solution S is clear (2.2.1) and not more intensely coloured Sulfated ash (2.4.14)
than reference solution BY, (2.2.2, Method II). Maximum 0.1 per cent, determined on J.O g.
Test solution Dissolve 50.0 mg of the substance to be water Rand 75 volumes of ethanol (96 per <en!> R. Using
examined in the mobile phase and dilute to 100.0 mL with o. J mL of phenolphthalein solution R as indicator, titrate with
the mobile phase. 0.1 M sodium hydroxide until a pink colour is obtained. Carry
Reference solution (a) Dilute 10.0 mL of the test solution to out a blanktitration.
100.0 mL with the mobile phase. Dilure 5.0 mL of this I mL of 0.1 M sodium hydr«<ide is equivalent to 36.18 mg
solution to 100.0 mL with the mobile phase. of C 19H,oCINO-r-
IMPURITIES
Specified impuriues A, B, G, D, E.
Reference solution (c) To 1 mL of the test solution, add
I mL of 0.1 M hydnxhloric acid and evaporate to dryness on
a hot plate. Dissolve the residue in 20 mL of the mobile
phase.
Column:
=
- size: 1 0.125 m, 0 =
4 mmj
- stationary phase: octadecylsilYl Sl7ica gelfor chromatography R
A.4-chloro-N-[2-(4-hydroxyphenyl)ethyl]benzamide
(5~).
(chlorobenzoyltyramine),
Mobile phase Mix 40 volumes of a 2.72 gIL solution of
potassium dihydrogen phosphate R adjusted to pH 2.3 with C02H
phosphoric acidR, and 60 volumes of methanol R. r"Y
Flow rate I mUmin. Cl AJ
bifection 20 ~L. B. 4-chlorobenzoic acid,
Run time The time necessary to detect the ester, which,
depending on the route of synthesis, may be impurity CJ D
orE.
Re/aliveretention Wirh reference to bezafibrate (retention
= =
time about 6.0 min): impurity A about 0.5;
System suitobility: C. methyl 2-[4-[2-[(4-chlorobenzoyl)amino]ethyl]phenoxyJ-2-
- resolution: minimum 5.0 between the 2 principal peaksin methylpropanoate,
the chromatogram obtainedwith reference solution (c);
the chromatogram obtained with reference solution (b).
Limits:
- impurities A, B, C, D E: for each impurity, not more than
J

- unspecified impun·lies: for each impurity, not more than D. ethyl 2-[4-[2-[(4-chlorobenzoyl)amino]ethyl)phenoxy]-2-
0.2 times the area of the principal peak in the methylpropanoate,
(0.10 per cent);
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1-314 Bicalutamide 2022
Reference solution (b) Dissolve 5 mg of bicolutamide for system

suitabzlity CRS (containing impurities B and C) in the solvent
mixture and dilute to 5.0 mL with the solvent mixture.
Reference solution (c) Dissolve 25.0 mg of Mcalutamide CRS
mixture. Dilute 5.0 mL of the solution [Q 25.0 mL with the
E. butyl 2-[4-[2-[(4-chlorobenzoyl)aminojethyl]phenoxy]-2- solvent mixture.
methyJpropanoate. Column:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" - size: 1 = 0.25 m, 0 = 4.0 nun;
- stationary phase: spherical end-capped octadecylsilyl silica gel
for chromatography R (5 um);
Bicalutamide *** Mobile phase:
** ** - mobile phaseA: phosphon'c acid R, aatonitn1e Rt, waterR
(Ph. Bur. monograph 2196) ***** (1.9:100:1900 VIVIV);
- mobile phase B: phosphoric acid R, waterR, acetonitrile Rl
O
o,,: H'Y'H'_ ~ '(X CF'
(1.9:100:1900 ViVlV);
-?" I . . . . . - If ~ I and enanUomer
F~ 0 ~ eN (min) (per cent VIJI) (per cent VIJo')
0-3 92 S
430.4 90357-06-5 3 - 23 92 --> 61 8 --> 33
23 - 43 67 --> 50 33 --> 50
Action and use 43 - 50 50 50
Antiandrogen; treatment of prostate cancer.
Preparation Flow rale 1.0 mUmin.
Bicalutamide Tablets Detection Spectrophotometer at 210 run.
PIlE" _ Injection 10 J.IL of test solution (a) and reference
solutions (a> and (b).
DEFINITION
(2RS)-N-[4-Cyano-3-(trilluoromethyl)phenyl]-3-[(4- with bicalutamide for SYSlem suitability CRS and the
lIuorophenyl)sulfonyl]-2-hydroxy-2-methylpropanamide. chromatogram obtainedwith reference solution (b) to
Content identify the peaks due to impurities Band C.
97.5 per cent to 102.0 per cent (dried substance). Relative retention With reference to bicalutamide (retention
CHARACTERS =
time about 38 min): impurity B =about 0.98;
Appearance =
White or almostwhite powder. System suitability Reference solution (b):
Solubility - peak-to-1Jalley ratio: minimwn 2.5, where HI' = height
Practically insoluble in water, freely soluble in acetone, above the baseline of the peakdue to impurity B and
slightly soluble in anhydrous ethanol and in methylene H v = height above the baseline of the lowest point of the
chloride. curve separating this peakfrom the peak due to
bicalutamide.
I[ shows polymorphism (5.9).
Limits:
IDENTIFICATION - impun'/y C: not more than 1.5 times the area of the
Infrared absorption spectrophotometry (2.2.24). principal peak in the chromatogram obtained with
Comparison bi<alutamide CRS. reference solution (a) (0.15 per cent);
If the spectra obtained in the solid state show differences, - unspecified impunties: for each impurity, not more than the
dissolve the substance to be examined and the reference area of the principal peakin the chromatogram obtained
substance separately in acetone R, evaporate to dryness and with reference solution (a) (0.10 per cent);
record new spectra using the residues. - total: not more than 5 times the area of the principal peak
Related substances - disregard limit. 0.5 times the area of the principal peak in
Liquid chromatography (2.2.29). the chromatogram obtained with reference solution (a)
Solvent mixture phosphoric acid R, acetonitrile Rl, waterR (0.05 per cent).
(0.05:50:50 VIVIV).
Test solution (a) Dissolve 25.0 mg of the substance to be Maximum 0.5 per cent, determined on 1.000 g by drying in
examined in the solvent mixture and dilute to 25.0 mL with an oven at 105 "C for 4 h.
Sulfa<ed ash (2.4.14)
Ten solution (b) Dilute 5.0 mL of test solution (a) to Maximum 0.1 per cent, determined on 1.0 g in a platinum
25.0 mL with the solvent mixture. crucible.
Reference solution (a) Dilute 1.0 mL of test solution (a) [0
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2022 Bifonazole 1-315
CN
ASSAY
O "rj
,
o a
s~
\' q
'.
CHJ
a
~ D-;::Y
"" I
CF,
enc enanucmer
Injection Test solution (b) and reference solution (c). F OH

Calculate me percentage content of C18Hl4.F~204S taking
into account the assigned content of bicahuamide CRS. H. (2RS)-N-[4-qano-3-(trilluoromethyl)phenyl)-2-[(4-
f1uorophenyl)sulfonyl)-3-hydroxy-2-methylpropanamide.
IMPURlTIES
Specified impuniies C.
Otherdeuctabte impurities (thefollowing substances would, if
HXH~~'(XCF'
I s---- I( ,?" I
present at a sufficient level, be detected by om or other of the tests
O
F~
,;:P' and enanllomer
0::::::,... eN
criterion for other/unspecified impurities and/or l>y thegeneral J (2RS)-N- [4-cyano-3-( triIIuoromethyl)phenyl]-3- [(4-
monograph Substances for phQ/maceutical use (2034). II is f1uorophenyl)sulfanyl]-2-hydroxy-2-methylpropanamide,
therefore not necessary UJ idenu'jy these impurities for
in substances for pharmaceutical use) A, B, D, E, F, H, J, K,
L,M.
OOHOCH,~
?'" ,.~. ~ K. (2R,2' S)-3,3' -sulfonylbis[N-[4-cyano-3-(trilluoromethyl)
f"y S
0 Y'( CF3 and eneouomer phenyl)-2-hydroxy-2-methylpropanamide).
V ~CN
A. (2RS)-N-[4-cyano-3-(trilluoromethyl)phenyl]-2-hydroxy-
2-methyl-3-(phenylsulfonyl)propanamide.
andenantiomer
cx r
O"rXH,.~'(XCF'
I . . . . .- I andenanliomer L. (2RS,2'RS)-3,3' -sulfonylbis[N-[4-cyano-3-
~ F eN (trilluoromethyl)phenyl]-2-hydroxy-2-
methylpropanamide],
B. (2RS)-N-[4-cyano-3-(trilluoromethyl)phenyl)-3-[(2- o OHO CH3
f1uorophenyl)sulfonyl)-2-hydroxy-2-methylpropanamide,
0'S'0 CO, H and enanllomer
OOHCH,~
f"y"s"~ Y'(CF3 andenanUomer

F
M.(2RS)-3-[(4-f1uorophenyl)sulfonyl)-2-hydroxy-2-
F~ 0 ~CN methylpropanoic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PlrErr
C. (2RS)-N- [4-cyano-3-(triIIuoromethyl)phenyl]-3-[(4-
f1uorophenyl)sulfonyl)-2-methylpropanamide,
H,NY'(CF, Bifonazole ****

~CN *** *
(Ph. Bur. monograph 1395) *** *
D.4-amino-2-(trilluoromethyl)benzoniniJe,
1 HXH'.~'(XCF'
r
I' . . ,.,. .
and enanliomer
F D~
~!
eN
andenanllomer
310.4 606ZIJ.96-8
E. (2RS) -N- [4-cyano-3-(triIIuoromethy1)phenyl) -3-[ (RS) -(4-
f1uorophenyl)sulfinyl)-2-hydroxy-2-methylpropanamide. Action and use
Antifungal.
~ HOpH3 H
.s , .xI ~N'(XCF' PlrErr _
F D~
I ....., I
eN
and enanUomer
DEFINITION
1-[(RS)-(Biphenyl-4-yl)phenyhnethyl]-IH-imidazole.
F. (2SKJ-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-[(RS)-(4- Content
f1uorophenyl)sullinyl]-2-hydroxy-2-methylpropanamide, 98.0 per cent to 100.5 per cent (dried substance).
CHARACTERS
Appearance
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1-316 Bifonazole 2022
Solubility - impurities B, D: for each impurity, not more than 5 times

Practically insoluble in water, sparingly soluble in anhydrous the area of the principal peak in me chromatogram
ethanol. obtainedwith reference solution (a) (0.5 per cent);
It shows polymorphism (5.9). - impurities A, C: for each impurity, not more than twice the
IDENTIFICATION with reference solution (a) (0.2 per cent);
Infrared absorption specrrophctomerry (2.2.24). - impw;ty E: not more than 1.5 times the area of the
Comparison bif01lazoie CRS. principal peak in the chromatogram obtained with
If the spectra obtained in the solid state show differences, reference solution (a) (0.15 per cent);
dissolve the substance to be examined and the reference - unspecified impurities: for each impurity, not more than the
substance separately in the minimum volume of 2-propanol R, areaof the principal peak in the chromatogram obtained
evaporate to dryness and record new spectra using the with reference solution (a)' (0.10 per cent);
residues. - total: not more than 10 times the area of the principal
peak in the chromatogram obtainedwith reference
TESTS
Related substances - disregard limit: 0.5 times the area of the principal peak in
Buffer solution pH 3.2 Mix 2.0 mL of phosphoric acidR with (0.05 per cent).
980 mL of water R, adjust to pH 3.2 (2.2.3) with
'riethylamine R and dilute to 1000.0 mL with waterR.
Maximwn 0.5 per cent, determined on 1.000 g by drying in
Test solution Dissolve 50.0 mg of the substance to be an oven at 105 °C.
examined in 25 mL of ace"'nitrile R and dilute to 50.0 mL
with buffer solution pH 3.2.
100.0 mL with buffer solution pH 3.2. Dilute 1.0 mL of this ASSAY
solution to 10.0 mL with buffer solution pH 3.2. Dissolve 0.250 g in 80 mL of anhydrous acetic acidR. Titrate
Reference solution (b) Dissolve 2 mg of bif01lazole for system with 0.1 M perchloric acid, determining the end-point
suitability CRS (containing impurities A, B, CJ D and E) in potentiometrically (2.2.20).
2 mL of acetenitrile R and dilute to 10.0 mL with buffer I mL of 0.1 M perchloric acid is equivalent to 31.04 mg
solution pH 3.2. of C,2H,sN2'
Column: IMPURITIES
- size: 1:::; 0.125 m, 0 ::: 4.0 mm; Specifid impurities A, B, C, D, E.
- stationary phase: octade<ylsilyl silka gd for chroma",graphy R
~ " " H ('I

(5 pm);
Mobile phase:
- mobile phase A: a","'nitrile Rl, buffer solution pH 3.2
~ - OH
andenantiomef
(20:80 VIII);
- mobile phase B: buffer solution pH 3.2, aceumitrile Rl A. (RS)-(biphenyl-4-yl)phenylmethanol,
(20:80 VIII);

(min) (per cent V/J? (per cent VIP) andenantiomer
0-8 60 40
N '"
8 - 12 60 ---> 10 40 ..... 90 L NH
12·30 10 90
B. 4-[(RS)-(biphenyl-4-yl)phenyimethyl]-IH-imidazole,
Flow rate 1 mIlmin.
Injection 50~.
with bif01lazoie for system suitability CRS and the
C. lH-imidazole,
identify the peaks due to impurities A, B, C, D and E.
Relative retention With reference to bifonazole (retention
impurity B = about 0.7; impurity A = about 3.2;
System suitobility Reference solution (b):
impurity Band bifonazole.
Limits:
peak area of impurity C by 2; D.I,3-bis[(biphenyl-4-yl)phenylmethyl]-IH-imidazolium ion,
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2022 Biotin 1-317
Detection Allow to cool and spray with

4-dimethylaminocinnama/dehyde solution R; examine
immediately in daylight.
reference solution.
TESTS
Soludon S
E. 1,4-bis[(biphenyl-4-yl)phenyhnethyl)-IH-imidazole. Dissolve 0.250 g in a 4 gIL solution of sodium hydroxide R
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<r and dilute to 25.0 mL with the same alkaline solution.
Specific opdcal rotadon (2.2.7)
Biotin *** + 89 to + 93 (driedsubstance), determined on solutionS.
*** *** Related substances
(ph. Eur. monograph 1073) *** Liquid chromatography (2.2.29). Prepare the solutwns
immediately before use and keep protected from bright light.
Solvent mixture water R, acetonitrile R (50:50 VIp).
examined in the solventmixture usingsonication and dilute
to 50.0 mL with the solventmixture.
244.3 58-85-5 100.0 mL with the solvent mixture. Dilute 1.0 mL of tltis
Action and use
Vitamin. Reference solution (b) Dissolve 5 mg of biotin for system
suitability CRS (containing impurities A, C and E) in the
PhEI.I _ solventmixture and dilute to 5 mL with the solventmixture.
DEFINlTION Column:
5-[(3aS,4S,6aR)-2-0xohexahydro--IH-thieno(3,4-djimidazol- - size: / = 0.25 m, 0 = 4.6 mm;
4-yl]pentanoic acid. - stationary phase: end-capped octade<y/silyf silica gelfor
Content - temperature: 30 ·C.
Mobile phase:
CHARACTERS - mobile phase A: meihanesvlfonic acidR, acetonitrile Rl, water
Appearance for chromatography R (1:25:1000 VIV/V);
White or almost white, crystalline powderor colourless - mobile phase B: methanesulfonic acid R, water for
crystals. chromatography R, acetonitrile R1 (1:25:1000 VIV/V);
Solublllty
Very slightly soluble in water and in ethanol (96 per cent), Tim. Mobile phase A MobUe phase B
practically insoluble in acetone. It dissolves in dilute solutions
(min) (percent Vm (per cent pm
of alkali hydroxides. 0-5 95 5
5 - 20 95 --> 0 5 ---> 100
IDENTIFICATION 20 - 28 0 100
First idenu"jication: A.
Secondidentification: B. Flow rate 1.0 mUmin.
A. Infrared absorption spectrophotometry (2.2.24). Detection Spectrophotometer at 200 DID from 0 to 5 min
Comparison biotin CRS. and at 210 nm from 5 to 28 min.
B. TItin-layer chromatography (2.2.27). Inieaion 10 lI1..
Prepare the sofutwns immediately bifore use and keep protected Identification of impurities Use the chromatogram supplied
from bright fighe with biotin for system suitability CRS and the chromatogram
Test so/mUm Dissolve 5 mg of the substance to be examined obtainedwith reference solution (b) to identify the peaks due
in glacial acetic acid R and dilute to 10 mL with me same to impurities A, C and E.
solvent. Relative retention With reference to biotin (retention
Reference solution Dissolve 5 mg of biotin CRS in glacial time = about 12 min): bromide = about 0.2;
aceticacid R and dilute to 10 mL with the same solvent. =
impurity C about 0.25; impurity A about I.1; =
Plate TLC silica gelplate R (5 pm). =
impurity E about 1.3.
Mobr1e phase methanol R, g!<Uial acetic acid R, toluene R System suitability Reference solution (b):
(5:25:75 VIV/V). - resolution: minimum 1.5 between the peaksdue to biotin
and impurity A.
Applicatwn 10 lI1..
Drying In a current of warm air.
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1-318 Biotin 2022

- correction factor: multiply the peak area of impurity E by IN
Hf;HH.~CO,H
. 1\ andepimeratC'
o~ S H CH3
0.2; N
- for each impurity, use the concentration of biotin in H H
Limits: D. (2RS}-2-methyl-5-[(3aS,4S,6.R)-2-oxohexahydro-IH-
- ;mpuniies A, E: for each impurity, maximum 0.5 per cent; thieno[3,4-d]imidazol-4-yl]pentanoic acid)
- impun"ty C: maximum 0.2 per cent;
0.10 per cent;
- reporting threshold: 0.05 per cent, disregard the peakdue to
the bromide ion.
an oven at 105°C.
ASSAY
Suspend 0.200 g in 5 mL of dimethylfonnamide R. Heat until E. 5-[(3aS,4S,6aR)-I-benzyl-2-oxohexahydro-IH-thieno[3,4-
the substance has dissolved completely. Add 50 mL of djimidazol-4-yl]pentanoic acid and 5-[(3aS,4S,6aR)-3-
ethanol R and titrate with 0.1 M teuabutylammomum benzyl-2-oxohexahydro-l H-thieno[3,4-dj imidazol-4-yl]
hydroxide, determining the end-point potentiometricaUy pentanoic acid,
(2.2.211).
I mL of 0.1 M retraburylammonium hydroxide is equivalent to
24.43 mg of C IO H 16N2 0 , S.
STORAGE
IMPURITIES
Specified impu";oes A, C, E.
F. diethyl 4-[(3aS,4S,6aR)-1,3-dibenzyl-2-oxohexahydro-IH-
Other derecMble impurities (the following ",b'Mnces would, if
thieno[3,4-d]imidazol-4-yl] butane-I,I-dicarboxylate,
present at a sufficient level, be detected by oneor other of the tats
in the monograph. They are limited by the general Q«eprana
criterion for other/unspecified impumies and/or by thegeneral
monograph Substances for phannaceUlital use (2034). It is
in substances for pharmaceutical use) B) D) F) G) H.
o~
~f;H
H
." ,H
's~>=o
H
~ G. (3aR,8aS,8bS}-1,3-dibenzyl-2-oxodecahydrothieno
[I',2': 1,2]thieno[3,4-d]imidazol-5-ium,
N C~H S~N
H H H H
A. 5-[(3aS,4S,6aR)-2-oxohexahydro-l H-thieno[3,4-dj
imidazol-4-yl]-2-[[(3aS,4S,6aR)-2-oxohexahydro-lH-
thieno[3,4-djimidazol-4-yl]propyl]pemsooic acid,
o~
..
H
~f;H ~co,H
S CO,H
H.2-ethyl-5-[(3aS,4S,6.R)-2-oxohexahydro-IH-thieno[3,4-d]
imidazol-4-yl]pentanoic acid"
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'"
N
H H
B. 4-[(3aS,4S,6aR)-2-oxohexahydro-IH-thieno[3,4-dj
imidazol-4-yl]butane-I,1-dicarboxylic acid,
C. 5-[(2S,3S,4R)-3,4-diaminothiolan-2-yl]pentanoic acid,
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2022 Biperiden Hydrochloride 1-319

Biperiden Hydrochloride reference solution (a).
(Ph. Bur. monograph 1074) System suitability Referencesolution (b):
C. To about 20 mg add 5 mL of phosphoric acid R. A green
o and eneouomer • Hel

colour develops.
TESTS
Solution S
H Dissolve 0.1 0 g in carbtm dioxide-free water R, heating gently if
necessary, and dilute to 50 mL with the same solvent.
347.9 1235-82-1
Action and use Solution S is not more opalescent than reference
Anticholinergic. suspension IT (2.2.1) and is colourless (2.2.2, Method Il).
pH (2.2.3)
POE" _
DEFINITION Related substances
( IRS) -1- [( IRS,2SR,4RS)-Bicyclo[2.2.1 ]hept-5-en-2-yl)-I- Gas chromatography (2.2.28).
phenyl-3-(piperidin-I-yl)propan-l-ol hydrochloride. Test solution Dissolve 0"10 g of the substance to be
Content examined in methanol R and dilute to 10 mL with the same
99.0 per cent to 101.0 per cent (dried substance). solvent.
CHARACTERS Reference solution (a) Dilute 0.5 mL of the test solution to
Appearance 100 mL with methanol R. Dilute 10 mL of this solution to
White or almost white, crystalline powder. 50 mL with methanol R.
Solubility
Slightly soluble in water and in ethanol (96 per cent), very examined and 5 mg of biperiden impurity A CRS in
slightly soluble in methylene chloride.
methanol R and dilute to 5 mL with the same solvent. Dilute
1 mL of the solution to 10 mL with methanol R.
mp
Column:
About 280 QC, with decomposition,
IDENTIFICATION - size: 1= 50 m, (2) = 0.25 mm;
First identification: A, D. - stationary phase: pherr;I(5)methy/(95)polysiloxane R (film
Second identification: B, C, D. thickness 0.25 urn).
A. Infrared absorption spectrophotometry (2.2.24). Cardergas nitrogen for chromatagraphy R.
Comparison biperiden hydrochloride CRS. Flow ral< 0.4 mlJmin.
B. Thin-layer chromatography (2.2.27). Sp/itratio 1:250.
Test solution Dissolve 25 mg of the substance to be Temperature:
solvent. Time Temperature
(min) CC)
Reference rolution (a) Dissolve 25 mg of biperiden
Column 0-5 200
5 - 40 200 ..... 270
same solvent.
Injection port 250
Reference solution (b) Dissolve 5 mg of biperiden Detector 300
impurity A CRS in reference solution (a) and dilute to 2 mL
Pial< TLC silica gel Fm pial< R.
Injection 2 ~L.
Mobile phase diethylamine R, methanol R, toluene R
(1:1:20 VIVIV). Run time Twice the retention time of biperiden.
Application 5 ~L Relative retention With reference to biperiden: impurities
A, Band C =between 0.95 and 1.05.
System suitability:
Drying In air. - resolution: minimum 2.5 between the peak due to
Detection A Examine in ultraviolet light at 254 nm. biperiden (I" peak) and the peak due to impurity A
Results A The principal spot in the chromatogram obtained (2nd peak) in the chromatogram obtained with reference
with me test solution is similar in position and size to the solution (b);
principal spot in the chromatogram obtained with reference - signal-to-noise ratio: minimum 6 for the principal peak in
solution (a). the chromatogram obtained with reference solution (a).
Detection B Spray with dilul< potassium iodobisnluthal< Limirs:
solution R and then with sodium niu;re solution R and examine - impurities A, B" C: for each impurity, maximum
in daylight. 0.50 per cent of me area of the principal peak;
Results B The principal spot in the chromatogram obtained - any other impun"o/: for each impurity, maximum
with the test solution is similar in position, colour and size to 0.10 per cent of the area of the principal peak;
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1-320 Bisacodyl 2022
H~O
.: 0
- total 0/impurities A, Band C: maximum 1.0 per cent of
the area of the principal peak;
- total 0/ impun",ies other than A, Band C: maximum andenantiomer
0.50 per cent of the area of the principal peak; ~--

- disregard limit: 0.05 per cent of the area of the principal H
peak.
D. 1-[( lRS,2SR,4RS)-bicyclo[2.2.I]hept-5-en-2-yl]-3-
Impurity F (2.4.24)
(piperidin-I-yl)propan-I-one,
Maximum 2 ppm.
\-~
Sulfated ash (2.4.14) ,,~ 0 and enaeucrrer
Maximum 0.1 per cent, determined on 1.0 g. H
ASSAY E. 1-[(I RS,2RS,4RS) -bicyclo[2.2.I)hept-5-en-2-yl]-3-

Dissolve 0.200 g in 60 mL of ethanol (96 per «nO R. In a (piperidin-I-yl)propan-I-one,
closed vessel, titrate with 0.1 i....f alcoholic potassium hydroxide,
F. benzene.
determining the end-point potentiometrically (2.2.2Q).
___ ~ PIlE"
I mL of 0.1 M akaholic potassium hydroxide is equivalent to
34.79 mg of C2IH3<lCINO.
STORAGE
In an airtight container, protected from light. Bisacodyl
IMPURITIES
Specified impurities A, B, C, F. (ph. Bur. monogroph 0595)
in themonograph. They are limiUl! by the general acceptance
cmenon for other/unspecified impun"ties and/or by thegeneral
monogroph Substances for phonnoceuticol use (2034). It is
therefore not necessary w identify these impurities for
in substanas for pharmaceutical use) D, E. 603-50-9
361.4
~
H
Action and use
H-, ~ .. 0 andenanliomer
Stimulant laxative.
Preparations
BisacodyJ Suppositories
BisacodylGastro-resistant Tablets
H
PIlE" _
A. (IRS)-I-[( ISR,ZSR,4SR)-bicyclo[2.2.I]hept-5-en-2-yl)-I- DEFINITION

phenyl-3-(piperidin-I-yl)propan-I-ol (endo form), 4,4'-(Pyridin-2-ylmethylene)diphenyl diacetate,
Content
~
"I 0H 98.0 per cent to 101.0 per cent (dried substance).
H
-, H :
0 andenanliomer
CHARACTERS
Appearance
Solubility
H
Practically insoluble in water, soluble in acetone, sparingly
B. (IRS)-I-[( ISR,2RS,4SR)-bicyclo[2.2.I]hept-5-en-2-yl]-I- soluble in ethanol (96 per cent). It dissolves in dilute mineral
phenyl-3-(piperidin-I-yl)propan-I-ol, acids.
IDENTIFICATION
First identification: C.
o andenanliome'
Second identification: A, B, D.
B. Ultravioletand visible absorption spectrophotometry
(2.2.25).
H Test solution Dissolve 10.0 mg in a 6 gIL solution of
potassium hydroxide R in methonol R and dilute to 100.0 mL
C. (IRS)-I-[(IRS,2RS,4RS)-bicyclo[2.2.I]hept-5-en-2-yl]-I- with the same solution. Dilute 10.0 mL of this solution to
phenyl-3-(piperidin-I-yl)propan-I-ol, 100.0 mL with a 6 gIL solution of potassium hydroxide R in
methanol R.
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2022 Bisacodyl 1-321
Spearal range 220-350 run. Injection 20 ~L.

Absorptkm maximum At 248 run. Run time 3.5 times the retention time of bisacodyl.
Shoulder At 290 run. Identification 0/ impurities Use the chromatogram supplied
Specific absorbance at the absorption maximum 632 to 672. with bisacodyl for system suitabl1ity CRS and the chromatogram
C. Infrared absorption spectrophotometry (2.2.24). obtainedwith reference solution (b) to identify the peaks due
to impurities A, B, C, D and E.
Comparison bisacodyl CRS.
Relative retention With reference to bisacodyl (retention
If the spectra obtained in the solid state show differences, time e about 13 min): impurity A ;:; about 0.2;
dissolve the substance to be examined and the reference impurity B ;:; about 0.4; impurity C ;:; about 0.45;
substance separately in chlorofonn R, evaporate to dryness and impurity D = about 0.8; impurity E =about 0.9;
record new spectra using the residues. =
impurity F about 2.6.
D. Thin-layer chromatography (2.2.27). Systemsuicaln1ity Reference solution (b):
Test solution Dissolve 20 mg of the substance to be - peak-w-valley ratio: minimum 1.5, where Hp ;:; height
examined in acetone R and dilute to 10 mL with the same above the baselineof the peak due to impurity E and
solvent. H,,;:; height above the baseline of the lowest point of the
Reference solution Dissolve 20 mg of bisacodyl CRS in curve separating this peak from the peak due to bisacodyl.
acetone R and dilute to 10 mL with the same solvent. Limits:
Plate TLC silica gel GF", plate R. - correction factor. for the calculation of content, multiply the
Mobile phase methylethylluume R, xylene R (50:50 VIV). peak area of impurity A by 0.7;
- impun"lies A, B: for each impurity, not more than the area
Applicarion 10 ~L.
of the principal peakin the chromatogram obtained with
Development Ovec a path of 10 em. reference solution (a) (0.1 per cent);
Drying In air, if necessary heating at 100-105 "C. - impurities C, E: for each impurity, not more than 5 times
Detection Spray with a mixture of equal volumes of 0.05 M the areaof the principal peak in the chromatogram
iodine and dilute su!/uric acidR. obtained with reference solution (a) (0.5 percent);
Results The principal spot in the chromatogram obtained - impun·ty D: not more than twice the area of the principal
with the test solution is similar in position and size to the peak in the chromatogram obtained with reference
principal spot in the chromatogram obtainedwith the solution (a) (0.2 per cent);
reference solution. - impurity F: not more than 3 times the area of the principal
TESTS solution (a) (0.3 per cent);
Acidity or alkalinity - unspecified impun·tres: for each impurity, not more than the
To 1.0 g add 20 mL of carborl dioxide-free waterR, shake, area of the principal peak in the chromatogram obtained
heat to boiling, cool and filter. Add 0.2 mL of 0.01 M sodium with reference solution (a) (0.10 per cent);
hydroxide and 0.1 mL of methyl red solution R. The solution is - total: not more than 10 times the area of the principal
yellow. Not more than 0.4 mL of 0.01 M hydrochloric acid is peak in the chromatogram obtained with reference
required to change the colour of the indicator to red. solution (a) (1.0 per cent);
Related substances - disregard limit. 0.5 times the area of the principal peak in
Liquid chromatography (2.2.29). Prepare the solutions the chromatogram obtained with reference solution (a)
immediauly be/ore use. (0.05 per cent).
Solvent mixture giaciol acetic acidR, acetonihue R, water R Loss on drying (2.2.32)
(4:30:66 VIVIV). Maximum 0.5 per cent, determined on 0.500 g by drying in
Test solution Dissolve 50 mg of the substance to be an oven at 105°C.
examined in 25 mL of acetomtrile R and dilute to 50.0 mL Sulfated ash (2.4.14)
with the solventmixture. Maximum 0.1 per cent, determined on 1.0 g.
Reference solution (a) Dilute 1.0 mL of the test solution to ASSAY
100.0 mL with the solvent mixture. Dilute 1.0 mL of this Dissolve 0.300 g in 60 mL of anhydrous acetic acid R. Titrate
solution to 10.0 mL with the solvent mixture. with 0.1 M perehloric acid determining the end-point
Reference solution (b) Dissolve 2.0 mg of bisacody! for system potentiometrically (2.2.2l1).
suitability CRS (containing impurities A, B, C, D and E) in 1 mL of 0.1 M perch/one acid is equivalent to 36.14 mg
1.0 mL of acetonitrile R and dilute to 2.0 mL with the solvent of C22HI')N04 .
mixture.
Reference sdtuion (c) Dissolve 5.0 mg of bisacody! for peak STORAGE
identification CRS (containing impurity F) in 2.5 mL of Protected from light.
acetonitrile R and dilute to 5.0 mL with the solvent mixture. IMPURITIES
Column: Specified jmpun·lres A, B, C, D, E, F.
- size: 1;:; 0.25 m, 0 ;:; 4.6 nun;
HO ,?" OH
- sUltionary phase: end-eapped octadecylsilyl silica gelfor
Mobile phase "Iix 45 volumes of acetonitrile Rand
55 volumes of a 1.58 gIL solution of ammonium formate R
previously adjusted to pH 5.0 with anhydrous formic acid R.
FWw rate 1.5 mllmin.
Detection Spectrophotometer at 265 run. A. 4,4'-(pyridin-2-ylmethylene)diphenol,
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1-322 Bismuth Subcarbonate 2022
OH To 6.6 mL of solution S add 4 mL of nitric acid R and dilute

to 50 mL with water R.
and enanliomer Nitrates
To 0.25 g in a 125 mL conical flask, add 20 mL of water R,
0.05 mL of indigo carmine solution Rl and then, as a single
B. 2-[(RS)-(4-hydroxyphenyl)(pyridin-2-yl)methyl)phenol, addition but with caution, 30 mL of sulfuric acid R. Titrate
immediately with indigo cannine solution RJ until a stable blue
HO colour is obtained. Not more than n mL of the titrant is
required, n being the volume corresponding to 1 mg ofN03 .
Alkall and aIkallne-earth metals
To 1.0 g add 10 mL of waterRand 10 mL of acetic acid R.
Boil for 2 min, cool and filter. Wash me residue with 20 mL
C.4-[(RS)-(4-hydroxyphenyl)(pyridin-2-yl)methyI]phenyl
of water R. To the combined filtrate and washings add 2 mL
of dilute hydrochlori< acid Rand 20 mL of water R. Boil and
acetate,
pass hydrogen sulfide R through the boiling solution until no
D. unknown structure, further precipitate is formed. Filter, wash the residue with
water R, evaporate the combined filtrate and washings to
dryness on a water-bath and add 0.5 mL of sulfuric acid R.
Ignitegently and allowto cool. The residue weighs a
maximum of 10 mg.
Arsenic (2.4.2, Method A)
Maximum 5 ppm.
To 0.5 g in a distillation flask add 5 mL of water Rand
7 mL of su!filric acid R, allow to cool and add 5 g of reducing
mixture R and 10 mL of hydrrxhlori< acid R. Heat the
E. 2-[(RS)-[4-(acetyloxy)phenyI](pyridin-2-yl)methyl]phenyl
contents of the flask to boiling gradually over 15-30 min and
acetate,
continue heating at such a rate thatthe distillation proceeds
F. unknown structure. steadily until the volume in the flask is reduced by half or
__ ~ POE" until 5 min after the air-condenser has become full of steam.
It is important thatdistillation be discontinued before fumes
of sulfur trioxide appear. Collect the distillate in a tube
containing 15 mL of water R cooled in ice-water. Wash down
Bismuth Subcarbonate the condenser with water R and dilute the distillate to 25 mL
with the same solvent. Prepare the standard usinga mixture
Bismuth Carbonate of 2.5 mL of arsenic standard solution (1 ppm As) Rand
(ph. Bur. monograph 0012) 22.5 mL of water R.
POE" ~ _ Copper
Maximum 50 ppm.
DEFINITION
To 5 mL of solution S, add 2 mL of ammonia R and dilute
Content
to 50 mL with water R. Filter. To 10 mL of the filtrate add
80.0 per cent to 82.5 per cent of Bi (A, 209.0) (dried
1 mL of a 1 gIL solution of sodium diethytdithiocarbamate R.
substance).
The solution is not more intensely coloured than a standard
CHARACTERS prepared at the same time in the samemanner using a
Appearance mixture of 0.25 mL of copper standard solutio" (10 ppm Cu) R
White or almost white powder. and 9.75 mL of waterR instead ofiO mL of the filtrate.
Solublllty Lead
Practically insoluble in water and in ethanol (96 per cent), Maximum 20 ppm.
It dissolves with effervescence in mineral acids. Atomic absorption spectrometry (2.2.23, Method If).
IDENTIFICATION Tesl solution Dissolve 12.5 g in 75 mL of a mixture of
A. It gives the reaction of carbonates (2.3.1). equa! volumes of lead-free nitric acid R and water R. Boil for
B. It gives the reactions of bismuth (2.3.1). 1 min, cool and dilute to 100.0 mL with water R.
TESTS
Reference solutians Prepare the reference solutions using
appropriate quantities of lead standard solution and a
Solution S
37 per cent VIV solution of lead-free nitric acidR.
Shake 5.0 g with 10 mL of waterR and add 20 mL of nitric
acidR. Heat to dissolve, cool and dilute to 100 mL with Source Lead hollow-cathode lamp.
waterR. Wavelength 283.3 urn (depending on the apparatus, the line
at 217.0 urn may be used).
Solution S is not more opalescent than reference Atomization device Air-acetylene flame.
suspension II (2.2.1) and is colourless (2.2.2, M"hod If). Silver
Chlorides (2.4.4) Maximum 25 ppm.
Maximum 500 ppm.
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2022 Bismuth Subgallate 1-323
To 2.0 g add I mL of water Rand 4 mL of ninic acid R. than 0.15 mL of 0.1 M sodium hydroxide is required to
Heat gently until dissolved and dilute to 11 mL with water R. change the colour of the indicator to yellow.
Cool and add 2 mL of 1 M hydrochlori< acid. AUow to stand Chlorides (1.4.4)
protected from light for 5 min. Any opalescence in the Maximum 200 ppm.
solution is not more intense chan that in a standard prepared
To 0.5 g add 10 mL of dilute nitric acidR. Heat on a water-
at the same time in the same mannerusing a mixture of
bath for 5 min and filter. Dilute 5 mL of the filtrate to
10 mL of silver standard solution (5 ppm Ag) R, I mL of nitric
15 mL with water R.
acid R and 2 mL of 1 M hydrochlone acid.
Nitrates
an oven at 105°C. To 1.0 g add 25 mL of water R then 25 mL of a mixture of
2 volumes of sulfuric add Rand 9 volumes of water R. Heat
ASSAY at about 50 °C for 1 min with stirring and filter. To 10 mL
Dissolve 0.500 g in 3 mL of nim"c acid R and dilute to of the filtrate, carefully add 30 mL of sulJUric acid R.
250 mL with water R. Carry out the complexometric titration The solution is not more intensely brownish-yellow than a
of bismuth (1.5.11). reference solution prepared at the same time as follows: to
1 mL of 0.1 M sodium edetate is equivalent (Q 20.90 mg ofBi. 0.4 g of gallic acid R, add 20 mL of nitrate standard solution
STORAGE (100 ppm NOll Rand 30 mL of a mixture of 2 volumes of
ndfioic acid Rand 9 volumesof water R, then filter, to 10 mL
of the filtrate, carefully add 30 mL of sulJUric acid R.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ !'hE"
Copper
Maximum 50 ppm.
Bismuth Subgallate Test solution Solution S.
(ph. Eur. monograph 1493) copper standard solution (10 ppm Cu) R and diluting with a
Ho-a,
0=qeo,H
I
6.5 per cent VIVsolution of lead-free nitric acid R.
'0 --P Wavelength 324.7 nm.
OH A tomisauon device Air-acetylene flame.
Lead
394.1 99-26-3 Maximum 20 ppm.
!'hE" _ Atomic absorption spectrometry (2.2.13, Method11).
DEFINTIlON Test solution Solution S.
Complex of bismuth and gallic acid. Reference solutions Prepare the reference solutions using lead
Content standard solution (10 ppm Ph) R and diluting with a
48.0 per cent to 51.0 per cent of Bi (A, 209.0) (dried 6.5 per cent VIV solution of lead-free ni.ric acid R.
substance). Source Lead hollow-cathode lamp.
CHARACTERS Wavelength 283.3 nm (depending on the apparatus, the line
Appearance at 217.0 urn may be used).
Yellowpowder. Atomisauon device Air-acetylene flame.
Solubility Silver
Practically insoluble in water and in ethanol (96 per cent). Maximum 25 ppm.
It dissolves in mineral acids with decomposition and in Atomic absorption spectrometry (1.1.13, Method1).
solutions of alkali hydroxides, producing a reddish-brown Test solution Solution S.
liquid. Reference solutions Prepare the reference solutions using silver
IDENTIFICATION standard solution (5 ppm Ag) R and diluting with a
A. Mix 0.1 g with 5 mL of water Rand 0.1 mL of phosphoric 6.5 per cent VIVsolution of lead-free nitric acid R.
acid R. Heat to belling and maintain boilingfor 2 min. Cool Source Silverhollow-cathode lamp.
and filter. To the filtrate, add 1.5 mL offerne chloride Wavelength 328.1 urn.
solution Rl, a blackish-blue colourdevelops.
Atomisanon device Air-acetylene flame.
B. It gives reaction (b) of bismuth (1.3.1).
Substances not precipitated by ammonia
TESTS Maximwn 1.0 per cent.
Solution S In a porcelain or quartz dish, ignite 2.0 g, increasing the
In a porcelain or quam dish, ignite 1.0 g, increasing the temperature verygradually to 600 ± 50 °C; allow to cool.
temperature verygradually. Heat in a mufflefurnace at Moisten the residue with 2 rnL of nimc acidR, evaporate to
600 ± 50 °C for 2 h. Cool and dissolve the residue with dryness on a water-bath and carefully heat and ignite once
warming in 4 mL of a mixture of equalvolumes of lead-free more at 600 ± 50 °C. Aftercooling) dissolve the residue in
nitric acid R and waterR and dilute to 20 mL with water R. 5 mL of nimc acid R and dilute to 20 mL with water R.
Acidity To 10 rnL of this solution, add concentrated ammonia R until
Shake 1.0 g with 20 mL of waterR for I min and filter. alkaline and filter. Wash the residue with waltT Rand
To the filtrate add 0.1 mL of methyl red solution R. Not mote
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1-324 Bismuth Subnitrate 2022
evaporate the combined filtrate and washings to dryness on a Acldlty

water-bath. Add 0.3 mL of dilute sulfuric acidR and ignite. Suspend 1.0 g in 15 mL of water R and shakeseveral times.
The residue weighs a maximum of 10 mg. Allow to stand for 5 min and filter. To 10 mL of the filtrate,
Loss on drying (2.2.32) add 0.5 mL of phenolphthalein solution RI. Not more than
Maximum 7.0 per cent, determined on 1.000 g by drying in 0.5 mL of 0.1 M sodium hydroxide is required to change the
an oven at 105°C for 3 h. colour of the indicator to pink.
Chlorides (2.4.4)
ASSAY
Maximum 200 ppm.
To 0.300 g add 10 mL ofa mixture of equal volumes of
nitric acid R and water R, heat to boilingand maintain boiling To 5.0 mL of solution SI, add 3 mL of nitric acid Rand
for 2 min. Add 0.1 g of potassium chlorate R, heat to boiling dilute to 15 mL with water R.
and maintain boiling for I min. Add 10 mL of water Rand Copper
heat until the solution becomes colourless. To the hot Maximum 50 ppm.
solution) add 200 mL of waler Rand 50 mg of ;'CY1eno1 orange Atomic absorption spectrometry (2.2.23, Method1).
triturate R. Titrate with 0.1 1\-1 sodium edetau until a yellow Testsolution Solution S2.
colour is obtained.
1 mL of 0.1 1\01 sodium edetate is equivalent to 20.90 mg of Bi. copper standard solutum (10 ppm Cu) R and diluting with a
STORAGE 37 per cent VIV solution of lead-free nitric acid R.
Protected from light. Source Copper hollow-cathode lamp.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1IE<r
Wavelength 324.7 run.
Atomisation deoice Air-acetylene flame.
Lead
Maximum 20 ppm.
Heavy Bismuth Subnitrate Atomic absorption spectrometry (2.2.23, Merhod 11).
(Ph. Bur. monograph 1494) Test solution Solution S2.
4[BiNO,(OHh],BiO(OH) 1462 1304-85-4 Reference solutions Prepare the reference solutions using lead
P1IE<r _ standard solution (10 ppm Pb) R and diluting with a
37 per cent VIV solution of lead-free nitric acid R.
DEFINITION Soura Lead hoUow-cathode lamp.
Content Wavelength 283.3 run (depending on the apparatus, the line
71.0 per cent to 74.0 per cent ofBi (A, 209.0) (dried at 217.0 run may be used).
substance).
Atomisalion device Air-acetylene flame.
CHARACTERS
SUver
Appearance Maximum 25 ppm.
Atomic absorption spectrometry (2.2.23, Method 1).
Solubility
Testsolution Solution S2.
Practically insoluble in water and in ethanol (96 per cent).
It dissolves in mineral acids with decomposition. Reference solutions Prepare the reference solutions using silver
standard solution (5 ppm Ag) R and diluting with a
IDENTIFICATION 37 per cent VIV solution of lead-free nitric acid R.
A. Dilute I mL of solution SI (see Tests) to 5 mL with Source Silverhollow-cathode lamp.
water R and add 0.3 mL of potassium iodide solution R.
A blackprecipitate is formed which dissolves into an orange Wavelength 328.1 run.
solution with the addition of 2 mL of potassium iodide Aromisation device Air-acetylene flame.
solution R. Substances not precipitated by ammonia
B. It gives reaction (b) of bismuth (2.3.1). Maximum 1.0 per cent
C. It gives the reaction of nitrates (2.3.1). To 20 mL of solution SI, add concentrated ammonia R until
D. pH (2.2.3): maximum 2.0 for solution S2 (see Tests). an alkaline reaction is produced and filter. Wash the residue
with waterR, and evaporate the combined filtrate and
TESTS washings to dryness on a water-bath. To the residue, add
Solution SI 0.3 mL of dilute suljitric acid R and ignite. The residue weighs
Shake 5.0 g by gently heating in 10 mL of water R and add a maximum of 10 mg.
20 mL of nitric acid R. Heat until dissolution) cool and dilute
Maximum 3.0 pet cent, determined on 1.000 g by drying in
Solution S2 an oven at 105 "C.
Place 1.00 g in a 20 mL volumetric flask and add 2.0 mL of
lead-free nitric add R. Allow acid attack to take place without ASSAY
heatingand if necessary warm slightly at the end to Dissolve with heating 0.250 g in 10 mL of a mixture of
completely dissolve the test sample. Add 10 mL of water R, 2 volumes of perchlorn: acid Rand 5 volumes of water R.
sbake and add, in smaU fractions, 4.5 mL of lead-free To the hot solution, add 200 mL of water R and 50 mg of
ammonia R; shake and allow to cool. Dilute to 20.0 mL with xylenol orange triturate R. Titrate with 0.1 M sodium edetate
waterR) shakeagain and allow the solids to settle. The clear until a yellow colour is obtained.
supernatant solution is solution S2. I mL of 0.1 i\1 sodium edetau is equivalent to 20.90 mg of Bi.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1IE<r
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2022 Bismuth Subsalicylate 1-325
Atomisanon device Air-acetylene flame.

Bismuth Subsalicylate
Lead
(Ph. Eur. monograph 1495) Maximum 20 ppm.
C,H,BiO, 362.1 14882-18-9 Atomic absorption spectrometry (2.2.23, Method If).
POE" ~ _ Test solution SolutionS.
Reference solutions Prepare the reference solutions using lead
DEFINITION
standard ,oIution (10 ppm Pb) R and diluting with a
Complex of bismuthand salicylic acid. 6.5 per cent VIV solution of lead-free nitric acid R.
Content Source Lead hollow-cathode lamp.
56.0 per cent to 59.4 per cent ofBi (11,209.0) (dried
Wavelength 283.3 nm (depending on the apparatus, the line
substance).
at 211.0 nm may be used).
CHARACTERS Atomisation device Air-acetylene flame.
Appearance
Silver
Maximum 25 ppm.
Solubility
Atomic absorption spectrometry (2.2.21, Method /).
Practically insoluble in water and in alcohol. It dissolves in
mineral acids with decomposition. Test solution SolutionS.
Reference solutions Prepare the reference solutions using silver
IDENTIFICATION
rtandard solution (5 ppm Ag) R and diluting with a
A. To 0.5 g add 10 mL of hydrochloric acidRl. Heat on a 6.5 per cent VIV solutionof lead-free nitric acid R.
water-bath for 5 min. Cool and filter. Retain the filtrate for
Source Silver hollow-cathode lamp.
identification test B. Wash the residue with dilute hydrochloric
add R and then with waterR. Dissolvethe residue- in Wavelength 328.1 nm.
0.5-1 mL of dilutesodium hydroxide solution R. Add 15 mL of Atomisation device Air-acetylene flame.
waterR. Neutralise with dilute hydrochloric acidR. Soluble bismuth
The solution gives reaction (a) of salicylates (2.3.1). Maximum 40 ppm.
B. The filtrate obtained in identification test A gives Atomic absorption spectrometry (2.2.23, Method 1).
reaction (b) of bismuth (2.3.1).
Test solution Suspend 5.0 g in 100 mL of water R. Stir
TESTS constantly for 2 h at 20-23 QC. Filter through filterpaper
Solution S (slow filtration) then througb a cellulosemicropore
In a porcelain or quartz dish, ignite 1.0 gJ increasing the membrane filter (0.1 um), To 10.0 mL of clear filtrate, add
temperature very gradually. Heat in a mufflefurnace at 0.1 mL of nitric acidR.
600 ± 25°C for 2 h. Cool and dissolve the residue with Reference solutions Prepare the reference solutions using
wanning in 4 mL of a mixture of equal volumes of lead-free bismuth standard solution (100 ppm Bi) R and diluting with a
nitric acid R and water R and dilute to 20 mL with water R. mixture of equal volumes of dilute nitric add R and water R.
Acidity Source Bismuth hoUow-cathode lamp.
Shake 2.0 g with 30 mL of etherR for 1 min and filter. Wavelength 223.06 nm.
To the filtrate add 30 mL of afaJhol Rand 0.1 mL of thymol
Atomiunion demce Air-acetylene flame.
bluesolution R. Not more than 0.35 mL of 0.1 M sodium
hydroxide is required to change the colourof the indicator to Loss on drying (2.2.32)
blue. Maximum 1.0 per cent, determined on 1.000 g by drying in
an oven at 105 QC.
Chlorides (2.4.4)
Maximum 200 ppm. ASSAY
Dissolve 0.250 g in a mixtureof 2 mL of nioic acid R, 5 mL Dissolve with heating 0.300 g in 10 mL of a mixture of
of waterRand 8 mL of methanol R. 2 volumes of perdJloric acid Rand 5 volumes of water R.
To the hot solution, add 200 mL of water Rand 50 mg of
Nitrates
xylenol orange triturate R. Titratewith 0.1 M sodium edaau
until a yellow colouris obtained.
To 0.1 g add 10 mL of waterR and, with caution, 20 mL of
I mL of 0.1 M sodium edetate is equivalent to 20.90 mg ofBi.
sulfun·, acid R and stir. The solution is not more intensely
yellowcoloured than a reference solutionprepared at the STORAGE
same time using 0.1 g ofsalicyl~ acid R, 6 mL of water R, Protected from fight,
4 mL of nitrate standard sotudon (100 ppm NO~ Rand _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POE..
20 mL of sulfuric acid R.
Copper
Maximum 50 ppm.
Test solution Solution S.
Reference solutions Prepare me reference solutions using
copper standard solution (10 ppm Cu) R and diluting with a
6.5 per cent VIV solution of lead-free nitric acid R.
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1-326 Bisoprolol Fumarate 2022
Bisoprolol Fumarate *** Mobile phase:

*** *** - mobile phase A: 10 gIL solution of phosphoric acid R;
(ph. Eur. monograph 1710) *** - mobile phase B: 10 gIL solution of phosphonc acid R in
acetonitrile R J;
HO'~ Thn,
(min)
MobUe phase A
(per cent VIII)
MobUe phase B
(per cent VIP)
0-4 95 5
co,H
4-8 95 --> 80 5 --> 20
8·15 80 20
15·34 80 --> 20 20 --> 80
and enanliomer
3<1 - 36 20 80
767 104144-21-2
Flow rale 1.0 mIJmin.
Action and use Delation Spectrophotometer at 225 nm.
Beta-adrencceptor antagonist. Injection 10 111-
Preparation Identification of impurities Use the chromatogram supplied
Bisoprolol Tablets with bisoproIDI for peak identification CRS and the
PhEIl _
identify the peaks due to fumaric acid and impurities A and
DEFINITION E; use the chromatogram supplied with bisoproIDI for sysurn
(2RS)-I-[4-[[2-(I-Methylethoxy)ethoxy]methyl]phenoxy]-3- suitability CRS and the chromatogram obtained with
[(I-methylethyl)amino]propan-2-o1 fumarate. reference solution (c) to identify the peak due to impurity G.
Content Relative retention With reference to bisoproJol (retention
99.0 per cent to 101.0 per cent (anhydrous substance). time = about 18 min): impurity A = about 0.5;
CHARACTERS
=
impurity G about 1.1; impurity E about 1.2. =
System su1·tobilliy Reference solution (c):
Appearance
Whiteor almost white, slightly hygroscopic powder.
- peak-UMJalley ratio: minimum 2.5, where Hp height =
above the baseline of the peak due to impurity G and
Solubility HfJ = height above the baseline of the lowest point of the
Very soluble in water, freely soluble in methanol. curve separating this peak from the peak due to
It shows polymorphism (5.9). bisoprolol.
Infrared absorption spectrophotometry (2.2.24). - impunty G: not more than 2.5 times the area of the
Comparison bisoprolol fumarate CRS.
If the spectra obtained in the solid stateshow differences, - impurity A: not more than 1.5 times the area of the
dissolve the substance to be examined and the reference principal peak in the chromatogram. obtained with
substance separately in methanol R, evaporate and dry the reference solution (a) (0.3 per cent);
residues at 60 °C at a pressure not exceeding 0.7 kPa and - impurity E: not more than the area of the principal peak in
record new spectra using the residues. the chromatogram obtained with reference solution (a)
Related substances - unspecified impurities: for each impurity, not more than
liquid chromatography (2.2.29). 0.5 times the area of the principal peak in the
Solventmixture acetonitrile Rt, waterfor chromatography R chromatogram obtained with reference solution (a)
(20:80 VIII). (0.10 per cent);
- total: not more than 2.5 times the area of the principal
peakin the chromatogram obtained with reference
- disregard hin,i: 0.25 times the area of the principal peak in
Reference solution (a) Dilute 1.0 mL of the test solution to the chromatogram obtained with reference solution (a)
100.0 mLwith the solvent mixture. Dilute 2.0 mL of this (0.05 per cent); disregard the peak due 10 fumaric acid.
Water (2.5.12)
Reference solution (b) Dissolve the contents of a vial of Maximum 0.5 per cent, determined on 1.000 g.
bisoproWlfor peak identification CRS (containing impurities A
and E) in 1.0 mL of the solventmixture. Sulfaled ash (2.4.14)
bisoprolol for system suitability CRS (containing impurity G) in ASSAY
1.0 mL of the solvent mixture. Dissolve 0.300 g in 50 mL of anhydrous acetic acidR. Titrate
Column: with 0.1 M perchioric acid, determining the end-point
- size: 1= 0.25 m, 0 = 4.6 mm; potentiometrically (2.2.20).
- stationary phase: octadecy/silyl silica gelfor chromawgraphy R 1 mL of 0.1 M perchlDric acid is equivalent to 38.35 mg
(5 urn); of C,oH..N,O'2'
- temperature: 20 ± 2 'C.
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2022 Bisoprolol Fumarate 1-327
STORAGE Oy;,°H
IMPURITIES
Specified impurities A, E, G.
P "'-
o-...../"'
I
o
.CCH,
CH NACH
'H
A CH3
a
and enanllomer
present at a sufficient level, be detected by oneor other of lhe tests F. (2RS)-2-[4-(2-isopropoxy-ethoxymethyl)phenoxy]-3-
it! the monograph. They arelimited fry the get/eral acceptance isopropylaminopropan-2-o1,
cntetion for otherhmspecified impurities and/orby the general
therefore not necessary co identify these impun"ties for
demonstration of compliance. StIJ also 5.10. Control of ;mpun"ties
in substances for pharmaceutical use) B, C, D, F, s; L, N,
Q, R, S, T, U.
G. (2RS)-I-[4-[[(2-isopropoxyethoxy)methoxy]methyl]
phenoxy]-3-isopropylaminopropan-2-ol,
A. (2RS)-I-(4-hydroxymethyl-phenoxy)-3-
isopropylaminopropan-2-ol,
H OH
('y0~~yCH3 K. 2-isopropoxyethyl 4-[[(2RS)-2-hydroxy-3-
('/ CH3 (isopropylamino)propyl]oxy]benzoate,
O~o~CH3 and enanliomer
B. (2RS)-I-isopropylamino-3-[4-(2-propoxy-
ethoxymethyl)phenoxyjpropan-2-01,
H OH L. 4-[[(2RS)-2-hydroxy-3-(isopropylamino)propylj
"" O~~yCH' oxyjbenzaldehyde,
~ CH3 H OH
('y0~~yCH'
'"
"" H OH
rJ CH,
O~~yCH, O~O"""""""'CH3 andeoaotiomer
CH,
N. (2RS)-I-[4-[(2-ethoxyethoxy)methyl]phenoxy]-3-
isopropylaminopropan-2-ol,
c. 1-[4-[4-(2-hydroxy-3-isopropylamino-propoxy)benzyl]
phenoxy]-3-isopropylaminopropan-2-o1, H pH H
('y0~NyCH,
rJ CH,
o~oc~ andeoaoliomer
Q. (2RS)-I-(isopropylamino)-3-[4-(2-methoxyethoxy)
methyljphenoxypropan-z-ol,
D.I-[4-[4-(2-hydroxy-3-isopropylaminopropoxy)
benzyloxylmethyl]phenoxyj-3-isopropylaminopropan-2-01,
R. (2RS)-I-(isopropylamino)-3-(4-methylphenoxy)propan-2-
01,
E. (EZl-[3-[4-(2-isopropoxy-ethoxymethyl)phenoxy]
allyl]isopropylamine, s. d-hydroxybenzaldehyde,
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1-328 Bleomycin Sulfate 2022
o
Solublllty
0--< CH3 Very soluble in water, slightly soluble in anhydrous ethanol,
D
O~N --< practically insoluble in acetone.
I CH,
OHC ~
IDENTIFICATION
A. Examine me chromatograms obtained in the test for
T.4-[(3-isopropyl-2-oxo-I,3-oxazoUdin-5-yl) composition.
methoxy]benzaldehyde, Results The 2 principal peaks in the chromatogram obtained
with the test solution are similar in retention time and size to
the 2 principal peaks in the chromatogram obtained with
B. It gives the reactions of sulfates (2.3.1).
TESTS
The solution is clear (2.2.1) and its absorbance (2.2.25) at
U. 5-[[4-(bydroxymethyl)phenoxy)methyl]-3-isopropyl-l,3-
oxazolidin-z-one.
______________ ~ PhE'I Dissolve 0.200 g in waterR and dilute to 10.0 mL with the
same solvent.
pH (2.2.3)
4.5 to 6.0.
Bleomycin Sulfate Dissolve 50 mg in ,arbondiaxide-free water R and dilute to
Bleomycin Sulphate Composition
(Ph. Bur. monograph 0976) Liquid chromatography (2.2.29); use the normalisation
procedure.
solvent.
Reference solution (a) Dissolvethe contents of a vial of
b1emnycin tulfate CRS in water R and dilute to 10.0 mL with
the same solvent.
Reference solutian (b) Dilute 1.5 mL of reference solution (a)
to 100.0 mL with waterR.
Column:
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped ocuuk<y/siIyl.mea gd for
,hromawgraphy R (7 1lIIl).
Mobile phase:
- mobile phase A: methanol R;
- mobile phase B: dissolve 0.960 g of sodium
pentonesulfonau R in 900 mL of acetic acid (4.8 gIL
C,H,O,), add 1.86 g of sodium edetare R, dilute to
1000 mL with the same solvent and adjust to pH 4.3 with
9041-93-4 ammonia Rj
Action and use
Cytotoxic antibacterial. (min) (per cent V/J? (per cent VII')
Preparadon 0·60 10 -J 40 90 -0 60
Bleomycin Injection 60· end 40 60
PhE'I _
DEFINITION
Sulfate of a mixture of glycopeptides produced by
Streptomyces vertidOus or by any other means; the 2 principal Injulion 20 J1L.
components of the mixture are N-[3-(dimethylsulfaniumyl) Run lime Until impurity D is eluted (about 80 min).
propyl]bleomycinamide (bleomycin A,) and N-[4- Relative retention With reference to bleomycin A2 :
(carbamimidoylamino)butyl)bleomycinamide (bleomycin B,). =
impurity D 1.5 to 2.5.
Potency System suitalnl.iy:
Minimum 1500 IU/mg (dried substance). - resolution: minimum 5.0 between the peaks due to
bleomycin A, (I" principal peak) and bleomycin B,
CHARACTERS
(2nd principal peak) in the chromatogram obtained with
Appearance
reference solution (a);
White or yellowish-white, very hygroscopic powder.
- signal-to-nlise ratio: minimum 20 for the principal peak in
the chromatogram obtained with reference solution (b);
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2022 Bleomycin Sulfate 1-329

2 per cent for the principal peak after 6 injections of
Limits:
- bleomycin A z: 55 per cent to 70 per cent;
- bleomycin B z: 25 per cent to 32 per cent;
- sum of bleomycin A z and B z: minimum 85 per cent;
- impun"ly D: maximum 5.5 per cent;
- sum of impun"ties otherthan D: maximum 9.5 per cent;
- disregard limil: 0.1 per cent of the total.
Maximum 3.0 per cent, determined on 50 mg by drying at
60°C at a pressure not exceeding 0.67 kPa for 3 h.
Less than 5 ill/mg, if intended for use in the manufacture of
parenteral preparations without a funher appropriate
ASSAY
Carry out the microbiological assay of antibiotics (2.7.2), B. N-[3-[(4-aminobutyl)amino]propyl]bleomycinamide
using the diffusion method. Use blMmydn sulfate CRS as the (bleomycin A,),
chemical reference substance.
STORAGE
In an airtight container, at a temperature of 2 °C to 8 "C.
If the substance is sterile) the container is also sterile and
tamper-evident.
IMPURITIES
Specified impuniies D.
Otherd'l<f:labie impurities (th'lol/qwingsubsrances would, if
present at a su.fficient leveJp be detected by oneor otherof the tests
in the monograph. They are limited by thegeneral aaeptanu
criterion lor otherhmspecified impurities and/or ry the general
monograph Substances lor phamtaceutical us, (2034). It is
therefore not neussary to identify these impun"ties for
in substances for pharmaceutical use) A.. B.. C.
c. N-[4-[[N-[4-(carbamimidoylamino)butyl]catbamimidoyI)
amino]butyl]bleomycinamide (bleomycin B.),
A. bleomycinic acid,
D. N-[3-(methylsulfanyl)propyl]bleomycinamide (S-
demethylbleomycin A,).
__________ ~ PhE",
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1-330 Boldine 2022
*** pH 8.50 ± 0.05 with diethylamine R and dilute to

Boldine
** ** 1000 mL with waterfor chromatography R;
**** * - mobile phase B: acetonitrile Rj
OH Tim, MobUe phase A Mobile phase B

(min) (per cent VIJI) (per cent VIJ.')
~
'CO "'" 0-2 85 15
2 - 38 85 -> 64 15 -> 36
H,CO "'" I "", 38 - 50 64 ---+ 20 36 -> 80
HO ; N 50·53 20 80
H I
CH,
321.4 476-7IJ.O Detection Spectrophotometer at 302 nm.
Injection 10 ~L.
Action and use Identification of impurities Use the chromatogram supplied
Antioxidant. with boldine for system suitability CRS and the chromatogram
PIlE" _ obtained with reference solution (c) to identify the peaks due
[Q impurities A, C and E.
DEFINITION
Relative retention With reference (0 boldine (retention
(6aS)-I, I0-Dimethoxy-6-methyl-5,6,6a,1-tetrahydro-4H- =
time about 23 min): impurity A about 0.25; =
dibenzo[de,g]quinoline-2,9-diol, of vegetable origin. =
impurity C about 0.6; impurity E about 1.6. =
Content System suitability Reference solution (b):
98.5 per cent to 100.5 per cent (anhydrous substance). - resolution: minimum 10.0 between the peaks due to
CHARACTERS impurity C and boldine.
Appearance Calculation of percentage contents:
White or almost whiteor yellowish) crystalline powder. - for each impurity, use the concentration of boldine in
Solubility reference solution (a).
Very slightly soluble in water, soluble in ethanol Limits:
(96 per cent) and in methylene chloride. It dissolves in dilute - impuniy C: maximum 1.5 per cent;
acid solutions. - impun'ty A: maximum 0.3 per cent;
- impun'cy E: maximum 0.15 per cent;
IDENfIFICATlON
0.10 per cent;
Comparison boldine CRS. - lOcal: maximum 1.5 per cent;
TESTS - reporting threshold: 0.05 per cent.
Specific optical rotation (2.2.7) 2-Propanol (2.4.24)
+ 121.0 to + 121.0 (anhydrous substance). Maximum 1.0 per cent.
Dissolve 0.500 g in ethanol (96 per cen!l R and dilute to Water (2.5.32)
50.0 mL with the same solvent. Maximum 0.5 per cent, determined on 0.100 g using the
Related substances evaporation techniqueat 120 "C.
Liquid chromatography (2.2.29). Sulfated ash (2.4.14)
Ten solution Dissolve 12.0 mg of the substance to be Maximum 0.1 per cent, determined on 1.0 g.
examined in 8 mL of methanol R using sonication and dilute ASSAY
Dissolve 0.200 g in 50 mL of glacial acetic add R. Titrate
Reference sotauon (a) Dilute 1.0 mL of the test solution to with 0.1 M petchlotic acid, determining the end-point
10.0 mL with methanol R. Dilute 1.0 mL of this solution to potentiometrically (2.2.20).
I mL of 0.1 M perchloric acid is equivalent to 32.14 mg of
Reference solution (b) Dissolve 12 mg of boldine for system CI9H2IN04'
suitability CRS (containing impurities C and E) in 8 mL of
methanol R using sonication and dilute to 10 mL with IMPURITIES
methanol R. Specified impurities A, C, E.
Reference solution (c) In order to prepare impurity A in situ, Otherdete<table impurities (thefollowing substances would, if
add 0.5 mL of strong hydrogen peroxide soluwn R to 5 mL of present at a sufficient levd, be detected by oneor other of the tests
reference solution (b) and stirfor 1 h. in the monograph. They arelimited by the general acceptance
cntenonfor other/unspecified impurities and/or by the general
Column:
- size: I = 0.25 m, 0 = 4.6 mrn;
therefore not necessary to idemify these impurities for
- statIOnary phase: end-copped extra-dense banded octy/sllyl silica
demonstration of compliance. See aha 5.10. Control of inrpuniies
gelfor chromatography R (5 pm);
in substances for pharmaceutical we) BJ D.
Mobile phase:
- mobl1e phase A: dissolve 1.54 g of ammonium acetate R in
950 mL of water for chromawgraphy R, adjust to
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2022 Borage Oil 1-331
OH Solubility
Practically insoluble in water and in el:hanol (96 per cent),
miscible with light petroleum.
Relative density
HO About 0.921.
Refractive Index
About 1.476.
A. (6S,6aS)-2, 9-dihydroxy-I,1 0-dimethoxy-6-methyl- IDENTIFICATION
5,6,6a,7-tettahydro-4H-dibenzo[de,g]quinoline N-oxide, Firs' identification: B.
Second idemiji<:ation: A.
~
'CO ;H A. Identification of fatty oils by thin-layer chromatography
(2.3.2).
H,CO,? """ I Results The chromatogram obtained is similar to the
HO' i N
H r" 0 B. Composition of fatty acids (see Tests).
C",
TESTS
Acid value (2.5.1)
B. (6R,6aS)-2,9-dihydroxy-I,10-dimethoxy-6-methyl- Maximum 0.5, or maximum 0.3 if intended for use in the
5,6,6a,7-tettahydro-4H-dibenzo[de,glquinoline N-oxide,
manufacture of parenteral preparations.
OH Peroxide value (2.5.5, Method A)
Maximum 10.0, or maximum 5.0 if intended for use in the
manufacture of parenteral preparations.
",CO Unsaponlliable matter (2.5.7)
HO
Alkaline impurities (2.4.19)
C. (6aS)-I, I0-dimethoxy-5,6,6a,7-tetrahydro-4H-dibenzo[de, Composition offatty acIds (2.4.22, Method A)
g]quinoline-2,9-diol, . Use the mixture of calibrating substances in Table 2.4.22.-3.
Composition of'he faUy-acidfraction of the oil:
- saturatedfaltY acidsofchain length less than C/6: maximum
0.3 per cent;
- palmitic acid: 9.0 per cent to 12.0 per cent;
- palm;UJleic acid: maximum 0.6 per cent;
HO - stearic acid: 2.0 per cent to 6.0 per cent;
- oleic acid: 12.0 per cent to 22.0 per cent;
- linoleic acid: 30.0 per cent to 41.0 per cent;
- gamma-Uno/mit; acid: 17.0 per cent to 27.0 per cent;
D. (6aS)-1,2,10-lrimethoxy-6-methyl-5,6,6a,7-tettahydro-4H- - alpha-linolenit; acid: maximum 0.5 per cent;
dibenzo[de,glquinolin-9-o1, - arachidic acid: maximum 0.5 per cent;
E. unknown structure. - eicosmoic acid: 2.8 per cent to 4.4 per cent;
_______________ ~ PIIEId - erode acid: maximum 3.0 per cent;
- nenxmic acid: maximum 4.5 per cent.
Water (2.5.32)
Refined Borage Oil STORAGE.
Under an inert gas, in a well-filled, airtight container,
Refined Startlower Oil
protected from light.
(Refined Borage (Starflower) Oil, Ph. Bur. monograph 2105)
PIIEId _ LABELLING
The label stases:
DEFINlTION - where applicable, that me oil is suitable for use in the
Fatty oil obtained from seeds of Borago officinalis L. manufacture of parenteral preparations;
by extraction and/or expression. It is then refined. A suitable - the name of the inert gas.
antioxidant may be added. ___________________ ~PhEId
PRODUCTION
The oil is prepared using materials and methods designed to
ensure that the content of brassicasterol (2.4.23) in the sterol
fraction of the oil is not greater than 0.3 per cent.
CHARACTERS
Appearance
Clear, light yellow or yellow liquid.
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1-332 Borax 2022
Borax *** Boric Acid

*** ***
Sodiwn Borate *** (Ph. Bur. monograph 0001)
Sodium Tetraborate H,BO, 61.8 10043-35-3
(Ph. Eur. monograph 0013) PhE<r _
Na,B,O"IOH,O 381.4 1303-96-4 DEFINITION

PhElI _
Content
DEFINITION 99.0 per cent to 100.5 per cent.
Disodium tetraborate decahydrare. CHARACTERS
Content Appearance
99.0 per cent to 103.0 per cent ofNa2B.107JlOH20. White or almost white, crystalline powder, colourless, shiny
plates greasy to the touch, or white or almostwhite crystals.
CHARACTERS
Appearance Solublllty
White or almost white) crystalline powder) colourless crystals Soluble in water and in ethanol (96 per cent), freely soluble
or crystalline masses, efflorescent. in boiling water and in glycerol (85 per cent).
SolubWty IDENfIFlCATlON
Soluble in water, very soluble in boiling water, freely soluble A. Dissolve 0.1 g by gently heating in 5 mL of methanol R,
in glycerol. add 0.1 mL of su/fun' acid R and ignite the solution.
The flame has a green border.
IDENTIFICATION
B. Solution S (see Tests) is acid (2.2.4).
A. To I mL of solution S (see Tests) add 0.1 mL of su/fum
acidR and 5 mL of me'hanol R and ignite.The flame has a TESTS
green border. Solution S
B. To 5 mL of solution S add 0.1 mL of phenolphthalein Dissolve 3.3 g in 80 mL of boiling distiUed water R, cool and
solution R. The solutionis red. On the addition of 5 mL of dilute to 100 mL with carbon dioxide-free water R prepared
glycerol (85 percent) R the colour disappears. from distilled waterR.
C. Solution S gives the reactions of sodium (2.3.1). Appearance of solution
TESTS
Solution S pH (2.2.3)
Dissolve 4.0 g in carbon diodde-free water R prepared from 3.8 to 4.8 for solution S.
distilled water R and dilute to 100 mL with the same solvent. SolubWty In ethanol (96 per cent)
Appearance of solution The solution is not more opalescent than reference
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). suspension II (2.2.1) and is colourless (2.2.2, Method II).
pH (2.2.3) Dissolve 1.0 g in 10 mL of boiling ethanol (96 percent) R.
9.0 to 9.6 for solution S. Organic matter
Sulfates (2.4.13) It does not darken on progressive heatingto dull redness.
Maximum 50 ppm, determined on solution S. Sulfates (2.4.13)
Use in this test 1.0 mL of acetic acid R. Prepare the standard Maximum 450 ppm.
using a mixture of 3 mL of sulfa,. standard solution Dilute 10 mL of solution S to 15 mL with dis.lled water R.
(10 ppm SO,) Rand 12 mL of distilled waterR. ASSAY
Ammonium (2.4.1) Dissolve 1.000 g with heating in 100 mL of water R
Maximum 10 ppm. containing 15 g of mannitol R. Titrate with 1 M sodium
Dilute 6 mL of solution S to 14 mL with water R. Prepare hydroxUk, using 0.5 mL of phenolphthalein solution R as
the standard using a mixture of 2.5 mL of ammonium indicator, until a pink colour is obtained.
standard solution (I ppm NH,) Rand 7.5 mL of water R. I mL of 1 M sodium hydroxide is equivalent to 61.8 mg
Calclum (2.4.3) ofH,BO,.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
Prepare the standard using a mixture of 6 mL of calcium
standard solution (10 ppm Co) R and 9 mL of distilled water R.
ASSAY Botulinum Toxin Type A for
Dissolve 20 g of mannitol R in 100 mL of water R, heating if
necessary, cool and add 0.5 mL of phenolphthalein solution R .Injection
and neutralise with 0.1 M sodium hydroxide until a pink (ph. Eur. monograph 2113)
colour is obtained. Add 3.000 g of the substance to be PhE<r _
examined, heat until dissolution is complete, cool, and titrate
with 1 lH sodium hydroxide until the pink colour reappears. DEFINITION
I mL of 1 M sodium hydroxide is equivalent to 0.1907 g Botulinum toxin type A for injection is a dried preparation
of Na2B407,lOH20. containing purified botulinumneurotoxin typeA, whichmay
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r be present in the form of a complex with haemagglurinins
and non-toxic proteins. Botulinum neurotoxin type A or its
haemagglutinin complex is prepared by a suitable purification
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2022 Botulinum Toxin Type A 1-333
process of the liquid supernatant from a broth-eulture of a Production of active toxin

suitable strain of Clostridium botulinum type A. A bacterial strain producing a high yield of active toxin, as
The purified complexes consist of several proteins and can be determined by an acute toxicity assay, is suitable. Seed lots
of various sizes. The largest complex (relative molecular mass demonstrate a capability of producing at least a minimum
of about 900 000) consists of a 150 000 relative molecular toxicity level appropriate for the manufacturing process and
mass neurotoxin, a 130 000 relative molecular mass non- scale.
toxic protein and various haemagglutinins ranging between MANUFACTURER'S REFERENCE PREPARATIONS
relative molecular mass 14000 and 43 000. The purified During development, reference preparations are established
toxin moiety is composed of only the same 150 000 relative for subsequent verification of batch consistency during
molecular mass neurotoxin as is found in the 900 000 . production and for control of the bulk purified toxin and
relative molecular mass neurotoxin complex, which is initially finished product. They are derived from representative
produced as a single chain and further cleaved (nicked) by batches of botulinum toxin type A that are characterised as
endogenous proteases into a fully active, disulfide-linked, described under Bulk Purified Toxin.
54 000 relative molecular mass light chain and a 97 000
The reference preparations are suitably characterised for their
relative molecular mass heavy chain. intended purpose and are stored in suitably sized aliquots
The preparation is reconstituted before use, as stated on the under conditions ensuring their suitability.
label.
BULK PURIFIED TOXIN
PRODUCTION C. botulinum type A strain is grown anaerobically, in suitable
GENERAL PROVISIONS media, from which cultures are selected for step-up
Production of the toxin is based on seed cultures, managed incubations under a suitably controlled anaerobic atmosphere
in a defined seed-lot system in which the ability to produce through the seed culture and bulk fermentation stages to
toxin is conserved. The production method must be shown allow maximwn production of toxin. The toxin is purified by
£0 yield consistently product of activity and profile suitable methods to remove nucleic acids and components
comparable £0 that of lots shown in clinical studies to be of likely to cause adverse reactions.
adequate safety and efficacy. Only a purified toxin mat complies with the following
The production method and stability of the finished product requirements may be used in the preparation of the final
and relevant intermediates are evaluated using the tests bulk. For each test and for each product, limits of acceptance
below. Such tests include the specific toxin activity per are established and each new purified toxin must comply
milligram of protein of purified toxin in an appropriate with these limits.
functional model of toxin activity and may be supported by
Residual reagents
tests confirming the presence of botulinum toxin type A, and,
Removal of residual reagents used in purification steps is
if appropriate, associated non-toxic proteins. confirmed by suitable limit tests or by validation of the
BACTERIAL SEED LOTS process.
A higWy toxigenic strain of C. botulinum of known toxin Nucleic acids
type A and confirmed absence of genes encoding other Removal of nucleic acids is confirmed by suitable limit tests
botulinum toxins (particularly botulinum toxin types B or by validation of the process.
and F), with known origin and history, is grown using
Immunological identity
suitable media. The bacterial strain, used for the master seed
The presence of specific type A toxin is confirmed by a
lot, shall be identified by historical records that include
suitable immunochemical method (2.7.1).
information on its origin and the tests used to characterise
the strain. These will include morphological, cultural, Specific activity
biochemical, genetic and serological properties of the strain. The specific activity is confirmed in a mouse model of
The master seed lot and the working seed lot, where toxicity or by in vivo/ex t1iw methods validated with respect
applicable, must be demonstrated to have identical profiles. to the LDso assay and expressed in mouse LDso units per
Only a seed 10{ that complies with the following requirements milligram of protein. Specific activity must not be less than
may be used. 1 x 108 mouse LDso units per milligram of protein for the
150 000 relative molecular mass neurotoxin and must not be
Identification
less than 1 x 101 mouse LDso units per milligram of protein
Each seed lot is identified as containing pure cultures of
for the 900 000 relative molecular mass neurotoxin complex.
c. botulinum type A bacteria with no extraneous bacterial or
fungal contamination. Protein
The total protein concentration is determined by a suitable
Microbial purity
method. An acceptable value is established for the product:
Each seed lot complies with the requirements for absence of
and each batch must be shown to comply with the limits.
contaminating micro-organisms, The purity of bacterial
cultures is verified by methods of suitable sensitivity. These Protein profile
may include inoculation into suitable media and examination Identity and protein composition are determined by
of colony morphology. polyacrylamide gel electrophoresis (2.2.31) under reducing or
non-reducing conditions or by other suitable physicochemical
Phenotypic parameters
methods such as size-exclusion chromatography (2.2.30),
Each seed lot must have a known fatty acid profile, sugar
comparing with suitable reference standards.
fermentation profile (glucose, lactose, mannose, etc.) and
proteolytic activity and must demonstrate relevant lipase, Total viable count
lecithinase and gelatinase activity. It complies with the limits approved for the particular
Genetic purity product.
Each seed lot must have information on the toxin gene FINAL BULK
sequence and comply with requirements for the absence of The final bulk is prepared by adding approved excipients to
other genes encoding other toxin serotypes. the bulk purified toxin. The solution is filtered through a
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1-334 Botulinum Toxin Type B 2022
bacteria-retentive filter. If human albumin is added, it The test may be repeated but when more than I test is
complies with the monograph Human albumin performed, the results of aU valid tests must be combined in
solution (0255). the estimate of potency.
FINAL LOT LABELLING
The final bulk is distributed aseptically into sterile, tamper- The label states:
evident containers. Uniformity of fill is verified during filling - the number of units of toxin per vial with a statement
and the test for uniformity of content (2.9.6) is not required. that units are product specific and not applicable to other
The containers are closed so as to prevent contamination. preparations containing botulinum toxin type A;
Only a final lot that is within the limits approved for the - the name and the volume of the diluent to be added for
particular product and is satisfactory with respect to each of reconstitution of the dried product
the requirements given below under Identification, Tests and _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
Assay may be released for use.
pH (2.2.3)
The pH of the reconstituted product is within ± 0.5
pH units of the limit approved for the particular product. Botulinum Toxin Type B for
Water
Not more than the limit approved for the particular product.
Injection
IDENTIFICATION PhElr _
The presence of botulinum toxin type A is confirmed by a
suitable immunochemical method (2.7.1). DEFINITION
TESTS Botulinum toxin type B for injection is a liquid preparation
Sterility (2.6. I) containing purified botulinum neurotoxin type B, which may
It complies with the test for sterility. be present in the form of a complex with haemagglutinins
and non-toxic proteins. Botulinum neurotoxin type B or its
Bacterial endotoxin. (2.6. 14)
haemagglutinin complex is prepared by a suitable purification
Less than 10 ill per vial.
process of the liquid supernatant from a broth-culture of a
ASSAY suitable strain of Cbmridium botulinum type B. Suitable
In accordance with the provisions of the European stabilisers may be added.
Convention for the Protection of Vertebrate Animals Used The toxin is present in its native form as a complex of
for Experimental and Other Scientific Purposes, tests must neurotoxin and non-toxin proteins and haemagglutinins with
be carried out in such a way as to use the minimum number a total relative molecular mass of approximately 700 000.
of animals and to cause the least pain, suffering, distress or The neurotoxin is synthesised by the bacterium as a single-
lasting harm, The LD50 assay is associated with severe chain polypeptide of approximately 150 000 relative
suffering of animals and manufacturers are strongly molecular mass that is activated during the fermentation
encouraged to develop and validate assays that will reduce process via a proteolytic cleavage (nicking) by endogenous
the number of animals used, or refine or replace the test proteases. The nicked protein is a fully active double-chain
procedure with the goal of promoting animal welfare. polypeptide consisting of a heavy chain (100 000 relative
The potency of the reconstituted product is determined by molecular mass) and a light chain (50 000 relative molecular
an LD50 assay in mice or by a method validated with respect mass), conoected by a disulfide bond.
to the LD50 assay. The potency is expressed in terms of the
PRODUCTION
LD50 for mice or relative to the reference preparation.
GENERAL PROVISIONS
For determination of the LDso, graded doses of the product Production of the toxin is based on seed cultures, managed
are injected Intraperitcneally into groups of mice and the in a defined seed-lot system in which the ability to produce
LOso is calculated by the usual statistical methods (5.3) from toxin is conserved. The production method must be shown
the mouse lethality in each group. A suitable reference to yield consistently product of activity and profile
preparation is assayed in parallel; the potency of the toxin is comparable to that of lots shown in clinical studies to be of
expressed relative to the reference or the value found for the adequate safety and efficacy.
reference is within suitable limits defined in terms of the
The production method and stability of the finished product
assigned potency.
and relevant intermediates are evaluated using the tests
After validation with respect to the LD 50 assay (reference below. Such tests include the specific toxin activity per
method), the product may also be assayed by other methods milligram of protein of purified toxin in an appropriate
that are preferable in terms of animal welfare, for example functional model of toxin activity and may be supported by
mouse bioassays using paralysis as the end-point, ex m'w tests confirming the presence of botulinum toxin type B, and)
assays using mouse phrenic nerve diaphragm, endopeptidase if appropriate, associated non-toxic proteins.
assays in vitro and cell-based assays.
BACTERIAL SEED LOTS
For alternative replacement methods the potency is A highly toxigenic strain of C. botulinum of known toxin
calculated with respect to a suitable reference preparation type B and confirmed absence of genes encoding other
calibrated in mouse LD50 units. botulinum toxins (particularly botulinum toxin types A
The estimated potency is not less than 80 per cent and not and F), with known origin and history) is grown using
more than 125 per cent of the stated potency. suitable media. The bacterial strain, used for the master seed
The confidence limits (P = 0.95) are not less than lot, shaU be identified by historical records that include
80 per cent and riot more than 125 per cent of the estimated information on its origin and the tests used to characterise
potency. the strain. These will include morphological, cultural,
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2022 Botulinum Toxin Type B 1-335
biochemical, genetic and serological properties of the strain. Specific activity

The master seed lot and the working seed lot, where The specific activity is confirmed in a mouse model of
applicable, must be demonstrated to have identical profiles. toxicity or by in oiootex vivo methods validated with respect
Only a seed lot that complies with the following requirements to the LDso assay and expressed in mouse LD 50 units per
may be used. milligram of protein. Specific activity must not be less than
Identification I x J 08 mouse LD so units per milligram of protein.
Each seed lot is identified as containing pure culrures of Protein
C. botulinum type B bacteria with no extraneous bacterial or The total protein concentration is determined by a suitable
fungal contamination. method. An acceptable value is established for the product
Microbial purity and each batch must be shown to comply with the limits.
Each seed Jot complies with the requirements for absence of Protein profile
contaminating micro-organisms. The purity of bacterial Identity and protein composition are determined by
cultures is verified by methods of suitable sensitivity. These polyacrylamide gel electrophoresis (2.2.31) under reducing or
may include inoculation into suitable media and examination non-reducing conditions or by other suitable physicochemical
of colony morphology. methods such as size-exclusion chromatography (2.2.30),
Phenotypic parameters comparing with suitable reference standards.
Each seed lot must have a known fatty acid profile, sugar Total viable count
fermentation profile (glucose, lactose, mannose, etc.) and It complies with the limits approved for the particular
proteolytic activity and must demonstrate relevant lipase, product
lecithinase and gelatinase activity.
FINAL BULK
Genetic purity The final bulk is prepared by adding approved excipients to
Each seed lot must have information on the toxin gene
the bulk purified toxin, The solution is filtered through a
genomic location and on the toxin gene sequence) and
bacteria-retentive filter. H human albumin is added, it
comply with requirements for the absence of other genes
complies with the monograph Human albumin
encoding other toxin serotypes.
solution (0255).
Production of active toxin
FINAL LOT
A bacterial strain producing a high yield of active toxin, as
The final bulk is distributed aseptically into sterile) tamper-
determined by an acute toxicity assay, is suitable. Seed lots
evident containers. Uniformity of fill is verified during filling
demonstrate a capability of producing at least a minimum
and the test for uniformity of content (2.9.6) is not required.
toxicity level appropriate for the manufacturing process and
The containers are closed so as to prevent contamination.
scale. .
Only a final lot that is within the limits approved for the
MANUFACTIJRER'S REFERENCE PREPARATIONS particular product and is satisfactory with respect to each of
During development, reference preparations are established
the requirements given below under Identification, Tests and
for subsequent verification of batch consistency during Assay may be released for use.
production and for control of the bulk purified toxin and
finished product. They are derived from representative pH (2.2.3)
batches of botulinum toxin type B that are characterised as The pH of the product is within ± 0.5 pH units of the limit
described under Bulk Purified Toxin. approved for the particular product
The reference preparations are suitably characterised for their IDENTIFICATION
intended purpose and are stored in suitably sized aliquots The presence of botulinum toxin type B is confirmed by a
under conditions ensuring their suitability. suitable immunochemical method (2.7.1).
BULK PURIFIED TOXIN TESTS
C. botulinum type B strain is grown anaerobically, in suitable Sterility (2.6.1)
media, from which cultures are selected for step-up It complies with the test for sterility.
incubations under a suitably controlled anaerobic atmosphere
through the seed culture and bulk fermentation stages to
Less than 10 ill per vial.
allow maximum production of toxin. The toxin is purified by
suitable methods to remove nucleic acids and components ASSAY
likely to cause adverse reactions. In accordance with the provisions of the European
Only a purified toxin that complies with the following Convention for the Protection of Vertebrate Animals Used
requirements may be used in the preparation of the final for Experimental and Other Scientific Purposes, tests must
bulk. For each test and for each product, limits of acceptance be'carried our in such a way as to use the minimum number
are established and each new purified toxin must comply of animals and to cause the least pam, suffering, distress or
with these limits. lasting harm, The LDso assay is associated with severe
suffering of animals and manufacturers are strongly
Residual reagents encouraged to develop and validate assays that will reduce
Removal of residual reagents used in purification steps is
the number of animals used, or refine or replace the test
confirmed by suitable limit tests or by validation of the procedure with the goal of promoting animal welfare.
process.
The potency of the product is determined by an LDso assay
Nucleic acids in mice or by a method validated with respect to the LD50
Removal of nucleic acids is confirmed by suitable limit tests assay. The potency is expressed in terms of the LDso for
or by validation of the process. mice or relative to the reference preparation.
Immunological Identity For determination of the LDso, graded doses of the product
The presence of specific type B toxin is confirmed by a are injected intraperitoneally into groups of mice and the
suitable immunochemical method (2.7.1).
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1-336 Bovine Serum 2022
LD 50 is calculated by the usual statistical methods (5.3) from Traceability of serum is maintained from the final container
the mouse lethality in each group. A suitable reference to the abattoir of origin (for blood collected from slaughtered
preparation is assayed In parallel; the potency of lite toxin is animals) or to the herd of origin (for blood collected from
expressed relative to the reference or the value found for the donor animals).
reference is within suitable limits defined in terms of the Further guarantee of the safety and quality of serum may be
assigned potency. ensured by the use of a controlled donor herd, Where serum
After validation with respect to the l.D50 assay (reference is obtained from such a herd, the animals are subjected to
method), the product may also he assayed by other methods regular veterinary examination to ascertain their health status.
that are preferable in terms of animal welfare, for example Animals introduced into the herd are traceable as regards
mouse bioassays using paralysis as the end-point, ex vivo source, breeding and rearing history. The introduction of
assays using mouse phrenic nerve diaphragm, endopeptidase animals into the herd follows specified procedures, including
assays in vitro and cell-based assays. defined quarantine measures. During the quarantine period
For alternative replacement methods the potency is the animals are observed and tested to establish that they are
calculated with respect to a suitable reference preparation free from all agents and antibodies from which the donor
calibrated in mouse LD50 units. herd is claimed to be free. It may be necessary to test the
animals in quarantine for freedom from additional agents,
The estimated potency is not less than 80 per cent and not
depending on factors such as infonnation available on their
more than 125 per cent of the stated potency.
breeding and rearing history. It is recommended that animals
The confidence limits (P = 0.95) are not less than
in the herd should not be vaccinated against bovine viral
80 per cent and not more than 125 per cent of the estimated
diarrhoea virus. Tests are carried out for any agent and/or
potency.
antibody from which the herd is claimed to be free.
The test may be repeated but when more than I test is Serum is obtained by separation of the serum from blood
performed, the results of aU valid tests must be combined in cells and clot under conditions designed to minimise
the estimate of potency. microbial contamination. Serum from a number of animals is
LABELLING pooled and a batch number is allocated to the pool.
The label states the number of units of toxin per vial with a Appropriate steps are taken to ensure homogeneity of the
statement that units are product specific and not applicable harvested material, intermediate pools and the final batch.
to other preparations containing botulinum toxin type B. Suitable measures (for example filtration) are taken to ensure
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE" sterility or a low bioburden. Before further processing, the
serum is tested for sterility or bioburden. General and
specific tests for viral contaminants are carried out as
described below.
A step or steps for virus inactivation/removal are applied to
Bovine Serum serum intended for production of immunological veterinary
(Ph. Eur. monograph 2262) medicinal products. Unless otherwise justified and authorised
for a particular medicinal product, a step or steps for virus
PIlE" _
inactivation/removal are applied to serum intended for
DEFINITION production of human and non-immunological veterinary
Liquid fraction of blood obtained from the ox (Bos taurus L.) medicinal products.
and from which cells, fibrin and clotting factors have been INACTIVATION
removed. The inactivation procedure applied is validated with respect
Different types of bovine serum are used: to a suitable representative range of viruses covering different
- adult bovine seJUm obtained at slaughter from cattle that types (enveloped, non-enveloped, DNA, RNA viruses).
are declared fit for human consumption; The optimal choice of relevant and model viruses depends
- calf serum obtained at slaughter from animals, fit for strongly on the specific inactivation/removal procedure;
human consumption, before the age of 12 months; representative viruses with different degrees of resistance to
- new-born calf sernm obtained at slaughter from animals the type of treatment must be included. Bovine viral
before the age of 20 days; diarrhoea virus must be included in the viruses used for
- foetal bovine serum obtained from normal foetuses from validation. Serum free from antibodies against bovine viral
dams fit for human consumption; diarrhoea virus is used in part or all of the validation studies.
- donor bovine serum obtained by repeated bleeding of donor For bovine serum intended for use in immunological
animals from controlled donor herds. veterinary medicinal products, for inactivation by gamma
This monograph provides a general qualityspecification for bovine irradiation a minimum dose of 30 kGy is applied, unless
serum. VanOus measures are applied during theproduction of otherwise justified and authorised.
booine serum aimedat obtaining a product that is acceptable as Critical parameters for the method of virus
regards viral safety. No single measure, nor the combination 0/ inactivation/removal are established and the parameters used
measures outlined be/ow can guarantee compleu viral safety but in the validation study are strictly adhered to during
they rather reduce the risk involvedin the use of serum in the subsequent application of the procedures to each batch of
manufat;ture of medicinal products. It is therefore necessary for the serum.
monufaaurerof a medicinal produa to take account of this when For inactivation by ganuna irradiation, critical parameters
choosing the semmfor a particular use by making a risk include:
assessment. - the temperature;
- packaging configuration;
PRODUCTION - distribution of dosimeters to assess the effective dose
All stages of serum production are submitted to a suitable received by the product whatever its position;
quality management system. - the minimum and maximum dose received.
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2022 Brimonidine Tartrate 1-337
QUAliTY CONTROL TESTS APPliED TO EACH IDENTIFICATION

BATCH A. The electrophoretic pattern corresponds to that for serum
A suitable sample size for each batch is established. Specific and is consistent with the type (foetal or other) of bovine
tests for viral contaminants are validated with respect to serum.
sensitivity and specificity. The cell cultures used for general E. Bovine origin is confirmed by a suitable immunochemical
tests for viral contaminants are shown to be sensitive to a method (2.7.1).
suitable range of potential contaminants. Control cells used
in the tests are cultivated, where relevant, with a bovine TESTS
serum controlled and inactivated as described in this Osmolality (2.2.35)
monograph. Serum free from antibodies to bovine viral 280 mosmollkg to 365 mosmollkg for foetal bovine serum
diarrhoea virus is required for validation of the effect of and 240 mosmol/kg to 340 mosmol/kg for other types.
antibodies on me detection limits for bovine viral diarrhoea Total protein (2.5.33)
virus. 30 mgrml, to 4S mwmL for foetal bovine serum and
Tests carried out on the hatch prior to treatment minimum 35 mg/mL for oilier types.
The following tests are carried out on the serum (before any Haemoglobin
virus inactivation/removal steps, where applicable). Maximum 4 mg/ml., determined by a validated method,
Tests for viral contaminants General tests supplemented by such as spectrophotometry.
specific tests are carried out. Bacterial endotoxins (2.6.14)
Genera/tests Validated tests are carried out by inoculation of Less than 10 IU/mL for donor bovine serum, less than
the serum on at least 2 distinct ceU lines, one of which is of 2S IU/mL for foetal bovine serum, less than 100 IU/mL for
bovine origin. The cell lines used are suitable for detecting oilier types.
haemadsorbing viruses such as bovine parainfluenza virus 3 Sterility (2.6.1)
and cytopathic agents such as bovine herpesvirus 1. It complies with the test. Use 10 mL for each medium.
Spedjic IeSIS for viral contaminants (if no' detected Iry generol Mycoplasmas (2.6.7)
tests), where relevant in view of the country of on"gin of the It complies with the test.
serum Bluetongue virus, bovine adenovirus, bovine
parvovirus, bovine respiratory syncytial virus, bovine viral STORAGE
diarrhoea virus, rabies virus and reovirus. Depending on the Frozen at -10°C or below.
country of origin, specific tests for other viruses may be LABELLING
needed. The animal health status of countries is defined by The label states:
the 'Office International des Epizooties' (DIE). - the type of serum;
For serum to be subjected to a virus inactivation/removal - where applicable, that the serum has been inactivated and
procedure, if evidence of viral contamination is found in any the inactivation method;
of the tests described above, the serum is acceptable only if - where the serum has been inactivated by gamma
the virus is identified and shown to be present in an amount irradiation, me target minimum dose of the irradiation
that has been shown in a validation study to be effectively procedure.
inactivated. ____________________ "'E'"
For serum that is not to be subjected to a virus
inactivation/removal procedure, if evidence of viral
contamination is found in any of the tests described above,
the serum is not acceptable. Brimonidine Tartrate ***
A test for bovine viral diarrhoea virus antibodies is carried ** **
out; an acceptance criterion for the titre is established taking (Ph. Eur. monograph 2760) *****
account of the risk assessment.
Composition The content of a suitable selection of the
following components is determined and shown [0 be within
the expected range for the type of serum: cholesterol, (J.-, JJ-
and y-globulin, albumin, creatinine, bilirubin, glucose, serum
aspartate transaminase (SAST, formerly SGOT - serum
gluramic-oxaloacetic transaminase), serum alanine
transaminase (SALT, fonnerly SGPT - glutamic-pyruvic 442.2 70359-46-5
transaminase), phosphorus, potassium, calcium, sodium and
Action and use
pH.
Alphaj-adrenoceptor agonist; treatment of hypertension.
Tests carried out on the batch post-treatment
PhE", _
If bovine viral diarrhoea virus was detected before virus
inactivation/removal, the following test for bovine viral DEFINITION
diarrhoea virus is carried out after virus inactivationJremovaJ. S-Bromo-N-(imidazolidin-2-ylidene)quinoxalin-6-amine
Testfor bovine viral diarrhoea virus A validated test for (2R,3R)-2,3-dihydroxybutanedioate.
bovine viral diarrhoea virus is carried out, for example by
Content
inoculation into susceptible cell cultures] followed by not
fewer than 3 subcultures and detection by immunostaining.
No evidence of the presence of bovine viral diarrhoea virus is CHARACTERS
found. Appearance
White or slightly yellowish or slightly brownish powder.
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1-338 Brimonidine Tartrate 2022
Solubility perchlotic add; determining the end-point potentiometrically

Soluble in water, practically insoluble in anhydrous ethanol (2.2.20).
and in toluene. I mL of 0.1 M perchloric acid is equivalent to 44.22 mg of
IDENTIFICATION C"H".BrN,06'
A. Specific optical rotation (see Tests). IMPURITffiS
B. Infrared absorption spectrophotometry (2.2.24). Otherdetectable impurities (thefollowing sub,tances would, if
Comparison bnmonidine tartrate CRS. present at a sufficient level.. be detected by one or other of the tests
in tm monograph. They are limiled by thegeneral a«ejJtance
TESTS criterion for other/unspecified impurities and/or I(y thegeneral
Specific optical rotation (2.2.7) monograph Sub"ances for pharmaceutical use (2034). It is
+ 9.0 to + 10.5 (dried substance). therefore not necessary to identify thes« impurities for
Dissolve 0.50 g in water R and dilute (Q 50.0 mL with the demon"..tion of compliance. See also 5.10. Control of impurities
same solvent. in substances for pharmaceutical use) A.. B.. CJ D.. EJ F.. G.
Related substances
Test sohuion Dissolve 65.0 mg of the substance (0 be
solvent.
100.0 mL with waterR. Dilute 1.0 mL of this solution to A N-(imidazolidin-2-ylidene)quinoxalin-6-amine.
Reference solution (b) Dissolve the contents of a vial of
brimonidine for system ,uitability CRS (containing impurity B)
in 1.0 mL of water R.
Column:
- size: 1= 0.25 rn, 0 = 4.6 mrn,
- stationary phase: end-capped octadecylsilyl sih'ca gelfor B. 5-bromoquinoxalin-6-amine,
chromarography R (5 urn); .
Mobile phase Dissolve 2.6 Ii of sodium heptanemlfonate R in
310 mL of methanol R, add 2.5 mL of niethylamine Rand
7.5 mL of glacial acetic acid R, and dilute to 1000 mL with
water R. Use a freshly prepared mixture.
F/qro rate 1.0 mIlmin. C. quincxalin-fi-amine,
Injection 20~.
Run lime 3 times the retention time of brimonidine.
Ilkntification of impurities Use the chromatogram supplied
with bnmonidine for system suitability CRS and the
D.l-(5-bromoquinoxalin-6-yl)thiourea,
identify the peak due to impurity B.
Relative retention Withreference to brimonidine (retention
= =
impurity E and brimonidine.
Calculation of percentage contents: B. 2-(5-bromoquinoxalin-6-yl)guanidine,
- for each impurity, use the concentration of brimonidine
tartrate in reference solution (a).
Limits: i~
- unspecified impuruies: for each impurity, maximum
0.10 per cent,
F. 5-bromo-N-(IH-imidazol-2-yl)quinoxalin-6-amine,
ASSAY
Dissolve 0.350 g in 70 mL of anhydrous acetic acid R using G.I-(2-aminoethyl)-3-(5-bromoquinoxalin-6-yl)urea.
sonication until complete dissolution. Titrate with 0.1 M _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
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2022 Bromazepam 1-339
Run time 4 times the retention time of bromazepam.

Bromazepam
Identification oj impurities Use the chromatogram supplied
(Ph. Eur. monograph 0879) with bromazepam for system suirabi/iOi CRS and the
chromatogram obtainedwith reference solution (b) to
identify the peaks due to impurities A, B, C, D and E.
Relative retention With reference to bromazepam (retention
time e about 5 min): impurity D : : : about l.4j
impurity A :::::: about 1.5; impurity C : : : about 1.6j
impurity E = about 2.1; impurity B = about 2.2.
System suilability Reference solution (b):
316.2 1812-3()'2 bromazepam and impurity D and minimum 1.2 between
the peaks due to impurities A and C.
Action and use
Benzodiazepine. Limits:
- correction [acton: for the calculation of content, multiply
Ph'" _ the peak areas of the following impurities by the
DEFINITION corresponding correction factor: impurity A : : : l.3j
7-Bromo-5 -(pyridin-2-yl)-1,3-dihydro-2H-I,4-benzod iazepin- impurity B : : : 1.8j impurity E : : : 2.1;
z-one. - impun'lies A, B, E: for each impurity, not more than the
Content with reference solution (a) (0.1 per cent),
CHARACTERS area of the principal peak in the chromatogram obtained
Appearance with reference solution (a) (0.10 per cent);
White or yellowish, crystalline powder. - total: not more than twice the area of the principal peak in
Solubility the chromatogram obtained with reference solution (a)
Practically insoluble in water, slightly soluble or sparingly (0.2 per cent);
soluble in ethanol (96 per cent) and in methylene chloride. - disregard limi,: 0.5 times the area of the principal peak in
IDENnFICATION (0.05 per cent).
Infrared absorption speotrophotcmetry (2.2.24).
Comparison bromozepom CRS. Maximum 0.2 per cent, determined on 1.000 g by drying at
TESTS 80°C at a pressure not exceeding 2.7 kPa for 4 h.
Related substances Sulfaled ash (2.4.14)
Liquid chromatography (2.2.29). Prepare the solutions Maximum 0.1 per cent, determined on 1.0 g.
immediately before we.
ASSAY
Test solution Dissolve 10.0 mg of me substance to be Dissolve 0.250 g in 20 mL of anhydrous acetic acid R.
examined in 9 mL of a mixture of 1 volume of acetonitrile R Add 50 mL of acetic anhydride R. Titrate with 0.1 M
and 8 volumes of methanol R. Dilute to 20.0 mL with an perchloric acid, determining the end-point potentiometrically
11.33 gIL solution of potassium dihydrogen phosphate R (2.2.20).
previously adjusted to pH 7.0 with a 100 gIL solution of
1 mL of O. 1 M perchloric acid is equivalent to 31.62 109
potassium hydroxide R.
of C 14HlOBrN30.
Reference sdution (a) Dilute 1.0 mL of the test solution to
100.0 mL with the mobile phase. Dilute 1.0 mL of this STORAGE
solution to 10.0 mL with the mobile phase. Protected from light.
Reference solution (b) Dissolve 5 mg of bromazepam for system IMPURITIES
suitability CRS (containing impurities A, B, C, D and E) in Specified impurities A, B, E.
5 mL of a mixture of 1 volume of acetonitrile Rand Other detectable impurities (thefollowing substances would, if
8 volumes of methanol R. Dilute to 10.0 mL with an present at a sufficient level, be detected by one or other of the tests
11.33 gIL solution of potassium dihydrogen phosphate R in the monograph. They are limited by thegeneral acceptance
previously adjusted to pH 7.0 with a 100 gIL solution of criterion for otherlunspecified impuriues and/or by thegeneral
potassium hydroxide R. monograph Substances for pharmaceutical use (2034). It is
Column: therefore not n«essary to identify these impun'lies for
- size: 1:::::: 0.15 m, 0 : : : 4.6 mrrn demonstration of compliance. See also 5.10. Control of impurities
- sraaonary phase: end-capped ocrade<ylsi/y/ silica gelfor in substances for pharmaceutical use) C, D.
chromaUJgraphy R (3.5 um);'
- temperature: 50 'C.
~
N H,
Mobile phase Mix 5 volumes of acetonitrile R, 45 volumesof

methanol Rand 50 volumes of an 11.33 gIL solution of B, "" I 0
potassium dihydrogen phosphate R previously adjusted to

N'"
pH 7.0 with a 100 gIL solution of potassium hydroxide R.
Flow rate 1.0 mlJmin. ""I
A. (2-amino-5-bromophenyl)(pyridin-2-yl)methanone,
Injection 20 ~L.
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1-340 Bromhexine Hydrochloride 2022
~
OI:.:' CI Content
CHARACTERS
B'
'" I 0
Appearance
N'"
Solubility
'" Very slightly soluble in water, slightly soluble in ethanol
B. N-[4-bromo-2-(pyridin-2-ylcarbonyl)phenyl]-2-
chloroaceramide,
IDENTIFICATION
~ 0 First idenu]kation: A, C.
Sr'
..- I
--N
-> Second idenrificau'on: B, C.
A. InIrared absorption spectrophotometry (2.2.24}.
Comparison bromhexine hydro<hloride CRS.
If the spectra obtained in the solid stateshow differences,
dissolve the substance (0 be examined and the reference
substance separately in methanol R, evaporate (0 dryness and
C. 7-bromo-5-(6-methylpyridinC2-yl)-1,3-dihydro-2H-1 ,4- record new spectra using the residues.
benzodiazepin-2-one, B. Thin-layer chromatography (2.2.27).
solvent.
Reference solution Dissolve 10 mg of bromhexine
hydrochloride CRS in methanol R and dilute to 5.0 mL with
the same solvent.
Plate TLC sih'ca gel F 254 plate R.
MobIle phase conantrated ammonia R, 2-propanol R,
D.3-amino-6-bromo-4-(pyridin-2-yl)quinolin-2(1H)-one, amone R (1:29:70 VIVIV).
Apph'calion 2 tJL.
°Y'ar Devdopmem Over 3/4 of the plate.
~
NH
Drying In air.
Br' 0 Detection Examine in ultraviolet light at 254 nm.
N'" with the test solution is similar in position and size to the
'" reference solution.
E. 2-bromo-N-[4-bromo-2-(pyridin-2-ylcarbonyl)phenyl] C. Dissolve about 20 mg in I mL of methanol R and add
acetamide. 1 mL of water R. The solution gives reaction (a) of chlorides
____________________ 1""" (2.3./).
TESTS
Bromhexine Hydrochloride *** than reference solution Y. (2.2.2, Method 11).
**• *** Dissolve 0.6 g in methanol R and dilute to 20 mL with the
(Ph. Bur: monograph 0706) *** same solvent. .
Related substances
Blf/fer solution Dissolve 1.:\6 g of ammonium formau R in
• He!
950 mL of water for chromatography R and adjust to pH 4.4
with anhydrous formic acid R. Dilute to 1000.0 mLwith water
for chromatography R.
Solvent mixrure aaumilrilt forchromarography R, water for
412.6 611-75-6 chromatography R (50:50 VIV).
Test solurion (a) Dissolve 50.0 mg of the substance to be
Action and use examined in the solvent mixture and dilute to 10.0 mL with
Mucolytic. the solvent mixture.
PM, _ Test solurion (b) Dilute 1.0 mL of test solution (a) to
DEFINITION
2,4-Dibromo-6-[[cyclohexyl(methyl)amino]methyl]aniline
hydrochloride.
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2022 Bromhexine Hydrochloride 1-341
Reference solution (aJ Dissolve 10 mg of bromhexine for system in the monograph. They arelimited by the general acceptance
su;tabl1ity CRS (containing impurities C and D) in the solvent criterion for otherlunsp«ified impurities and/or by the general
mixture and dilute to 2.0 mL with the solvent mixture. monograph Substances for phannaceutical use (2034). It is
Reference solution (b) Dilute 1.0 mL of test solution (a) to therefore not n«essary 10 identify these impumiesfor
100.0 mL with the solvent mixture. Dilute 1.0 mL of this demonstration of compliance. See also 5.10. Control of impun'ties
solution to 10.0 mL with the solvent mixture. in substances for pharmaceutical use) A, B, D, E.
Reference solution (c) Dissolve 50.0 mg of bromhexine
hydrochloride CRS in the solvent mixtuse and dilute to
10.0 mL with the solvent mixture. Dilute 1.0 mL of the
Column:
- size: /:::: 0.15 m, 0 :::: 2.1 mm;
A. (2-amino-3,5-dibromophenyl)methanol,
- srationary phase: end-capped solidme rxtadecy/si/y/ silica gel
for chromatography R (2.6 pm); CHO
- temperature: 30°C. ~NH,
i\1obile phase acetonitrile for chromatography R, buffer solution
(40:60 VIV). BrMsr
B. 2-amino-3J5-d.ibromobenzaldehyde,
Inj«tion 3.0 t-tL of test solution (a) and reference
Run time Twice the retention time of bromhexine.
with bromhexine for system suirabi/ilY CRS and the
chromatogram obtained with reference solution (a) to
identify the peaks due to impurities C and D. C. 2-[[cyclohexyl(methyl)amino)methyl]aniline,
Relative retention With reference to bromhexine (retention
=
impurity D about 0.3 -.
- resolution: minimum 2.0 between me peaks due to
impurities C and D.
Calculation of percentage amtents:
- correction faaor: multiply the peak area of impurity C by D.4-bromo-2-[[cyclohexyl(methyl)amino)methyl]aniline,
1.6;
- for each impurity, use the concentration of bromhexine
hydrochloride in reference solution (b). H'C.O
Limits:
- impuniy C: maximum 0.15 per cent; iN~ NH
and enanUomer
- unspecified impun'ties: for each impurity, maximum
-
0.10 per cent;
total: maximum 0.2 per cent;
OrDI '" Or
- reporting threshold: 0.05 per cent. E. (3RS)-6,8-dibromo-3-cyclohexyl-3-methyl-I,2,3,4-

Los. on drying (2.2.32) tetrahydroquinazolin-3-ium.
Maximum 1.0 per cent, determined on 1.000 g by drying in _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE..
an oven at 105 "C.
ASSAY
Injection Test solution (b) and reference solution (c).
Calculate the percentage content of Cl,.l{2IBr2C1N2 taking
into account the assigned content of bromhexine
hydrochloride CRS.
STORAGE
IMPURITIES
Specified impurities C.
Otherdetectable impurities (thefo/lJJwing subsumces wou/d,ij
present at a sufficient level, be detected by one or otherof me tests
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1-342 Bromocriptine Mesilate 2022
Specific absorbance at the absorption maximum 120 to 135

Bromocriptine Mesilate (dried substance).
(Ph. Eur. monograph 0596) B. Infrared absorption spectrophotometry (2.2.24).
Comparison bromoaiptine mesilate CRS.
C. Thin-layer chromatography (2.2.27). Prepare the solutions
immediately beJore use.
Solventmixture ethanol (96 per cenO R, methanol R, methylene
chloride R (30:30:40 VIVIV).
examined in the solvent mixture and dilute to 10 mL with
the solventmixture.
Reference solution Dissolve 10 mg of bromocriptine
C"H.,BrN,O,S 751 22260-51-1 mesilate CRS in the solvent mixture and dilute to 10 mL with
the solventmixture.
Action and use
Dopamine receptor agonist. Plate TLC silica gel G plate R.
Preparations lWobile phase concentrated ammonia R, water R, 2-propanol R,
Bromocriptine Capsules methylene chloride R, ether R (0.1:1.5:3:88:100 VIVIVIVIV).
Bromocriptine Tablets Applicaoon 10 pL.
Development Immediately in an unsaturated tank) over a
PhE" _
path of 15 em.
DEFINITION Drying In a current of cold air for 2 min.
(6aR,9R)-5-Bromo-N-[(2R,5S,10aS.l0bS)-IOb-hydroxy-2-(I- Deteaion Spray with ammonium molybdate solution R3 and
methylethyl)-5-(2-methylpropyl)-3,6-dioxooctahydro-8H- dry at 100·C until the spots appear (about 10 min).
oxazolo [3,2-a] pyrrolo [2, l-e1pyrazin- 2-yl]-7-methyl- Results The principal spot in the chromatogram obtained
4,6,6a,7,8,9-hexahydroindolo[4,3-jg]quinoline-9-carboxamide with the test solution is similar in position, colour and size to
monomethanesulfonate. the principal spot in the chromatogram obtained with the
Content reference solution.
98.0 per cent to 101.0 per cent (dried substance). D. To 0.1 g add 5 mL of dilute hydrochlcn"c acid R and shake
PRODUCTION for about 5 min. Filter and add I mL of banum chloride
It is considered that alkyl methanesulfonate esters are solution Rl. The filtrate remains clear. To a further 0.1 g add
genotoxic and are potential impurities in bromocriptine 0.5 g of anhydrous sodium carbonate RJ mix and ignite until a
mesilate. The manufacturing process should be developed white residue is obtained. Allow to cool and dissolve the
taking into consideration the principles of quality risk residue in 7 mL of waterR (solution A). Solution A gives
management, togetherwith considerations of the quality of reaction (a) of sulfates (2.3.1).
Slatting materials, process capability and validation. E. Solution A obtained in identification test D gives
The general methods 2.5.37. Merhyl, erhyl and isopropyl reaction (a) of bromides (2.3.1).
methanesulfonate in methanesulfonic acid, 2.5.38. Methyl, ethyl TESTS
and isopropyl methanesulfonate in acove rub,wnces and 2.5.39.
Methanesulfonyl chloride in meihanendfomc add areavailable to The solutionis clear (2.2.1) and not more intensely coloured
assist manufacturers. than reference solution Bj , BYj or Y", (2.2.2, Method 11).
CHARACTERS Dissolve 0.25 g in methanol R and dilute to 25 mL with the
Appearance same solvent.
White or slightly coloured, fine crystalline powder.
pH (2.2.3)
Solubility 3.1 to 3.8.
Practically insoluble in water, freely soluble in methanol, Dissolve 0.2 g in a mixture of 2 volumes of methanol R and
soluble in ethanol (96 per cent), sparingly soluble in 8 volumes of carbon dioxide-free water Rand dilute to 20 mL
methylene chloride. with the same mixture of solvents.
It is verysensitive to light.
The identification, tests and assay are 10 be carried out as rapidly + 95 to + 105 (dried substance).
as posoble, protected from lighc
Dissolve 0.100 g in a mixture of equal volumes of methanol R
IDENfIFICATION and methylene chloride R and dilute to 10.0 mL with the same
First identification: B. mixture of solvents.
Second identification: A, C, D, E. Related substances
A. Ultraviolet and visible absorption spectrophotometry Liquid chromatography (2.2.29).
(2.2.25). Solvent miXture buffer solution pH 2.0 R, methanol R
Test solution Dissolve 10.0 mg in 10 mL of methanolRand (50:50 VIV).
dilute to 200.0 mL with 0.01 M hydrochloric acid. Test solution Dissolve 0.500 g of the substance to be
Spectral range 250-380 run. examined in 5.0 mL of methanol R and dilute to 10.0 mL
Absorption maximum At 305 run. with buffer solution pH 2.0 R.
Absorption minimum At 270 run. Reference solution (a) Dilute 1.0 mL of the test solution to
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2022 Bromocriptine Mesilate 1-343
Reference solution (b) Dilute 1.0 mL of reference solution (a) IMPURITIES

to 10.0 rnL with the solvent mixture. Specified impumies A, B, C, D, E, F, G"
Reference solution (c) Dissolve me contents of a vial of
bromocriptine mesilate for system suitability CRS (containing
impurities A and B) in 1.0 mL of the solvent mixture. B, H CH,
I ~CH'
HN~",.(~;-N
: N 0 CH3
Column: -= .H
. 0
- size: 1;;;; 0.12 m, 0 ;;;; 4 mm;
- stationary phase: ocuuJet:yIsi/yl sili<a gelfor chromalCgraphy R
(5 urn).
I .#
H . O-\)N
0 .:
H,C\
Mobile phase: CH,
- mobile phase A: 0.791 gIL solution of ammonium
carbonate R; A. (6aR,9R)-5-bromo-7-methyl-N-[(2R,5S)-2-(I-
- mobile phase B: acetonitrile R; methylethyl) -5-(2-methylplopyl)-3,6-dioxo-2 ,3,5,6,9,10-
hexahydro-8H-oxazolo [3,2-a]pyrrolo [2,l-e] pyrazin-2-yl]-
Thn. Mobile phase A MobUe phase B 4,6,6a,7,8, 9-hexahydsoindolo[4, 3-jgJ quinoline-9-
(min) (per cent VIJI) (per cent JIll? carboxamide (z-bromcanhydro-c-ergocrypdne),
0-30 90 ..... 40 10 -+ 60
30 - 45 .0 60
Flow rou 2 mUmin.

Inje<tion 20 ~L.
with broTtJocripline mesilate for system suitability CRS and the
chromatogram obtained with reference solution (c) to identify B. (6aR,9R)-N-[(2R,5S,lOaS,10bS)-IOb-hydroxy-2-(I-
the peaks due to impurities A and B. methylethyl)-5-(2-methylpropyl)-3,6-dioxooclahydro-8H-
Re/alive retention With reference to bromocriptine: oxazolo [3,2-a] pyrrolo[2, I-c]pyrazin-2-yl]-7-methyl-
=
impurity C about 1.2. 4,6,6a,7,8, 9-hexahydsoindolo[4,3-fg] quinoline-9-
Systemsuitability Reference solution (c): carboxamide (e-ergocryptine),
impurities A and B.
Limits:
- impun'ty A: not more than 0.2 times the area of the
- impun"ty C: not more than 4 times the areaof the
reference solution (b) (0.4 per cent};
- impun"ties B, D, E, F, G: for each impurity, not more than C. (6aR,9S)-5-bromo-N-[(2R,5S, 10aS,IObS)-1 Ob-hydroxy-2-
twice the area of the principal peak in the chromatogram (l-methylethyl)-5-(2-methylpropyl)-3,6-dioxooctahydro-
obtained with reference solution (b) (0.2 per cent) and 8H-oxazolo[3,2-a]pyrrolo [2,l-e]pyrazin-2-yl)-7-me thyl-
not more than 1 such peak has an area greater than the 4,6,6a,7,8, 9-hexahydsoindolo[4, 3-fg] quinoline-9-
area of the principal peak in the chromatogram obtained casboxamide «8S)-2-bromo-«-ergoCl)lptine),
(0.05 per cern), apart from the peak due 10 impurity A.
Loss on drying (2.2.32) D. (6aR,9R)-5-bromo-7-methyl-4,6,6a,7,8,9-hexahydsoindolo
Maximwn 3.0 per cent, determined on 0.500 g by drying in [4,3-fg]quinoline-9-calboxylic acid,
ASSAY
Dissolve 0.500 g in 80 mL of a mixtute of 10 volwnes of
anhydrous acetic acid Rand 70 volumes of acetic anhydride R.
Titratewith 0.1 M perchloric acid, determining the end-point
1 mL of 0.1 M perchloric acid is equivalent 10 75.1 mg
of C"H"BrN5 0 SS. E. (6aR,9R)-5-bromo-7-methyl-4,6,6a,7,8,9-hexahydsoindolo
[4,3-fg]quinoline-9-calboxamide,
STORAGE
In an airtight container, protected from light, at a
temperature not exceeding -15°C.
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1-344 Bromperidol 2022
Br H?H, r:-- CH,

HN>=ctNl.(~H.0 N~~H'
0~~ o---k:~,
solvenr.
0
Reference solution (aJ Dissolve 10 mg of bromperidol CRS in
H3C OHU methanol R and dilute to 10 mL with the same solvent.
CH,
Reference solution (b) Dissolve 10 mg of bromperidol CRS
F. (6aR,9R)-5-bromo-N-[(2S,5S, IOaS,1ObS)-1Ob-hydroxy-2- and 10 mg of haloperirM CRS in melhanol R and dilute to
(l-methylethyl)-5-(2-methylpropyl)-3,6-dioxooctahydro- 10 mL with the same solvent.
8H-oxazolo [3,2-a[pyrrolc[2, I-e]pyrazin-2-yl]-7-methyl- Ptote TLC octadecylsilyl silica gel plate R.
4,6,6a,7,8,9-hexahydroindolo[4,3-fg]quinoline-9- Mobile phase lelYahydrofuran R, methanol R, 58 gIL solution
carboxamide «2'S)-2-bromo-«-ergocryptine), of sodium chloride R (10:45:45 VIVIV).
Application 1 ~L.
Development In an unsaturated tank, over 3/4 of the plate.
Drying In air.
System suitabifity Reference solution (b):
- the chromatogram shows 2 Spots which may, however)
not be completely separated.
G. (6aR,9R)-5-bromo-N-[(2R,5S,IOaS,IObS)-IOb-methoxy- Results The principal spot in the chromatogram obtained
2-(I-methylethyl)-5-(2-methylpropyl)-3,6-dioxooctahydro- with the test solution is similar in position and size to the
8H-oxazolo [3,2-a]pyrrolo [2, l-c]pyrazin-2-yl]-7-methyl- principal spot in the chromatogram obtained with reference
4,6,6a,7,8, 9-hexahydroindolo[4,3-fg] quinoline-9- solution (a).
carboxamide (2-bromo-lO 'b-D-methyl-a.-ergocryptine). D. Dissolve about 10 mg in 5 rnL of anhydrous ethanol R.
____________________ 1'11,,, Add 0.5 mL of dinitrobenzerul solution R and 0.5 rnL of 2 M
akoholic potassium hydroxide R. A violet colour is produced
that becomes brownish-red after 20 min.
E. To 0.1 g in a porcelain crucible add 0.5 g of anhydrous
Bromperidol sodium tarbonate R. Heat over an open flame for 10 min.
Allow to cool. Take up me residue with 5 mL of dilute nitric
(Ph. Eur. murwgraph 1178) acid R and filter. To 1 rnL of the filtrate add I rnL of
water R. The solution gives reaction (a) of bromides (2.3.1).
FY'lQOBr
~N I
TESTS
The solutionis clear (2.2.1) and not more intensely coloured
o
than reference solution Y7 (2.2.2, Melhod1I).
OH
Dissolve 0.2 g in 20 mL of a 1 per cent VIV solutionof lactic
<4, H,,BrFNO, 420.3 10457-90-6 acid R.
Related substances
Action and use Liquid chromatography (2.2.29).
Dopamine receptor antagonist; neuroleptic. Test solution Dissolve 0.100 g of the substance to be
I'll'" _ examined in methanol R and dilute to 10.0 rnL with the same
solvent.
DEFINITION
Reference solution (a) Dissolve 2.5 mg of bromperidcl CRS
4- [4-(4-Bromophenyl)-4-hydroxypiperidin-l-yl]-I-(4-
and 5.0 mg of haloperidol CRS in methanol R and dilute to
f1uorophenyl)butan-l-one.
50.0 rnL with the same solvent.
Content Reference solution (b) Dilute 5.0 rnL of the test solution to
99.0 per cent to 101.0 per cent (dried substance). 100.0 rnL with methanol R. Dilute 1.0 rnL of this solution to
CHARACTERS 10.0 rnL with methanol R.
Appearance Column:
White or almost white powder. - size: 1= 0.1 m, 0 ;;;; 4.0 mm;
Solubility - stationary phase: base-deactivated octadecy1si1y1 silica gelfor
Practically insoluble in water, sparingly soluble in methanol chrumawgraphy R (3 urn).
and in methylene chloride, slightly soluble in ethanol Mobife phase:
(96 per cent). - mobile phase A: 17 gIL solution of tetrabutylammonium
hydrogen sulfat< R;
IDENTIFICATION
- mobile phase B: auwnitrile R;
First identification: B, E.
A. Melting point (2.2.14): 156 -c to 159 'C.
Comparison bromperidol CRS.
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2022 Bromperidol 1-345
Time
(min)
0- 15
15·20
Mobile phase A
(per cent VIJ1
90
50
--> 50
MobUe phase B
(per cent VIV)
10 --> 50
50
r'lQ0Br
~N
F 0
I
0'
20 - 25 , 90 10 OH
B. 4- [4-(4-bromophenyl)-4-hydroxypiperidin-I-yl]-I-(2-
fluorophenyl)butan-I-one,
Injection 10 ~L.
Relativeretention With reference to bromperidol (retention
F~
10'
N
,I
impurity B = about 0.8; haloperidol = about 0.9; o
impurity C = about 1.4; impurity D = about 1.5; OH
impurity E = about 1.8; impurity F = about 1.85.
System suitability Referencesolution (a): C. 4-[4-(biphenyl-4-yl)-4-hydroxypiperidin-l-yl]-I-(4-
- resolution: minimum 3.0 between the peaks due to ftuorophenyl)butan-l-one j
haloperidol and bromperidol.

Limits:
- ;mpun"lies A., B.) CJ D, Ej F: for each impurity, not more
than the area of the principal peak in the chromatogram
obtained with referencesolution (b) (0.5 per cent);
- unspecified impurities: for each impurity, not more than
chromatogram obtained with reference solution (b) D. 4-[4-(4-bromophenyl)-4-hydroxypiperidin-l-yl]-I-(3-.,thyl-
-
(0.10 per cent);
total: not more than twice the area of the principal peak in
the chromatogram obtained with referencesolution (b) Br
4-ftuorophenyl) butan-f -one,
-
(1 per cent);
disregard limit: 0.1 times the area of the principal peak in
the chromatogram obtained with referencesolution (b)
(0.05 per cent).
6
an oven at 105°C.
HobY'lQ0B' ~N I
Sulfated ash (2.4. [4) o 0'
Maximum 0.1 per cent, determined on 1.0 g in a platinum OH
crucible.
E. 4-[4-(4-bromophenyl)-4-hydroxypiperidin-l-yl)-I-[4-[4-(4-
ASSAY
bromophenyl)-4-hydroxypiperidin-l-yl]phenyl)buran-l-
Dissolve 0.300 g in 50 mL of a mixture of I volume of one,
anhydrous acetic acid Rand 7 volumes of methylethylketone' R.
Titrate with O. l M perchloric acid, using 0.2 mL of
naphtholbenzei'n solution R as indicator.
"" Br
of C21H23BrFN02'
STORAGE OH
IMPURITIES F. 4-[4-(4 '~bromobiphenyl-4-yl)-4-hydroxypiperidin-l-yl)-I
Specified impuriues A, B, C, D, E, F. (4-lluorophenyl)butan-l-one.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'IIE,;
A. 1-(4-lluorophenyl)-4-(4-hydroxy-4-phenylpiperidin-I-yl)
buran-J-one,
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1-346 Bromperidol Decanoate 2022
Bromperidol Decanoate **** Mobile phase:

** * - mobile phaseA: 27 gIL solution of tetrabulylammonium
(Ph. Eur. monograph /397)
***.* hydrogen sulfate R;

(min) (per cent VIP) (per cent VIP)
0·30 80 ---040 20 ---060
30 - 35 4n eo
35 - 40 40 -+ 80 60 ---> 20

574.6 75067-66-2 Daeaion Spectrophotometer at 230 om.
Inj«tion I 0 ~L.
Action and use
Dopamine receptor antagonist, neuroleptic. Relativeretention With reference to bromperidol decenoare
(retention time = about 24 min): impurity G = ahout 0.10;
PhE" ~ _
=
impurity L ahout 0.15; impurity H ahout 0.8; =
DEFINITION impurity A =about 0.89; impurity I =about 0.91;
4-(4-Bromophenyl)-I-[4-(4-ftuorophenyl)-4- =
impurity B about 0.96; haloperidol
oxobutyl]piperidin-4-yl decanoate. decanoate = about 0.98; impurity F == about 1.10;
Content
=
impurity C about 1.15; impurity K about 1.2; =
impurity E = about 1.23; impurity D = about 1.25.
SYSl<m suitability Reference solution (a):
CHARACTERS - resolution: minimum 1.5 between the peaks due to
Appearance haloperidol decanoate and bromperidol decanoare.
White or almost white powder. Limits:
SOlubility - impurities A} B, C} D, E, P, G} H, I} J, K: for each
Practically insoluble in water, very soluble in methylene impurity, not more than the area of the principal peak in
chloride, soluble in ethanol (96 per cent). the chromatogram obtained with reference solution (b)
mp (0.5 per cent),
About 60 'C. - unspecified impurities: for each impurity, not more than
IDENTIFICATION chromatogram obtained with reference solution (b)
A. Infrared absorption spectrophotometry (2.2.24). (0.10 per cent);
Comparison bromperidol d«anOOl< CRS. - total: not more than 3 times the area of the principal peak
B. To 0.1 g in a porcelain crucible add 0.5 g of anhydrous in the chromatogram obtained withreference solution (b)
sodium carbonate R. Heat over an open flame for 10 min. (1.5 per cent);
Allow to cool. Take up the residue with 5 mL of dilute nitri, - disregard limir. 0.1 times the area of the principal peak in
acid R and filter. To I mL of the filtrate add I mL of the chromatogram obtained with reference solution (b)
water R. The solution gives reaction (a) of bromides (2.3.1). (0.05 per cent),
TESTS Loss on drying (2.2.32)

Appearance of solution Maximum 0.5 per cent, determined on 1.000 g by drying in
vacuo at 30 "C,
than reference solution B, (2.2.2, Method II). Sulfated ash (2.4.14)
Dissolve 2.0 g in methylene chloride R and dilute 10 20 mL Maximum 0.1 per cent, determined on 1.0 g in a platinum
with the same solvent. crucible.
Liquid chromatography (2.2.29). Prepare the solutions Dissolve 0.450 g in 50 mL of a mixture of I volume of
immediately before useand proteafrom lighe anhydrous acetic acid Rand 7 volumes of methylethyl ketone R.
Test solution Dissolve 0.100 g of me substance to be Titrate with 0.1 M perchloric acid, using 0.2 mL of
examined in methanol R and dilute to 10.0 mL with the same naphtholbenzein solution R as indicator.
solvent. I mL of 0.1 M perchloric acid is equivalent to 57.46 mg
Reference solution (a) Dissolve 2.5 mg of bromperidol of C3JH4LBrFN03'
deconoate CRS and 2.5 mg of haloperidol decanoale CRS in STORAGE
methanol R and dilute to 50.0 mL with the samesolvent. Protected from light, at a temperature below 25 "C.
Reference solution (b) Dilute 5.0 mL of the test solution to IMPURITIES
100.0 mL with methanol R. Dilute 1.0 mL of this solution 10 Specified impun'ties A, B} C, D, E, P, G, H, I, J, K.
Otherdetectable impurities (the foHowing substances would, if
Column: present at a sufficient level) be deuaed by oneor other of the rests
- size: 1= 0.1 ffi,0 = 4.0 mm; in the monograph. They are limited by the general acceptance
- stationary phase: base-deactiuated octade<y/si/yl silica gelfor criterion for otherlunspe<ified impurities andlor by thegeneral
chromawgraphy R (3 pm), monograph Substances for pharmaceutical use (2034). It is
therefore not necessary to identify these impunties for
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2022 Bromperidol Decanoate 1-347

in substances for pharmaceutical use) L.
F. 4-(biphenyl-4-yl)-I-[4-(4-fluorophenyl)-4-oxobutyl]
piperidin-4-yl decanoate,
A. 1-[4-( 4-fluorophenyl)-4-oxobu tylj-4-phenylpiperidin-4-yl

decanoate,
F'('laoB'
~N I
('laoB'
a
OH
~N I G. 4-[4-(4-bromophenyl)-4-hydroxypiperidin-I-ylj-I-(4-
Fa"" fluorophenyl)butan-l-one (bromperidol),
HJC~O
a
B. 4-( 4-bromophenyI}-I-[4-(2-fluorophenyl)-4-oxobutylj-
F'('lQ0B'
~N
a
I
piperidin-4-yl decanoate,
H3C~O
~""
a
I
N~ N
. B'
H,C H. 4-( 4-bromophenyl)-I-[4-(4-fluorophenyl)-4-oxobutylj
a ~ piperidin-sl-yl octenoate,
H3C~O
a
C. 4-( 4-bromophenyI}-I-[4-(3-ethyl-4-fluorophenyI}-4-
F~aoB'
oxobutyl]-piperictin-4-yl decanoate,
HJC~O
a
I. 4-(4-bromophenyl) -1- [4-(4-fluorophenyl)-4-oxobutyl]
piperidin-4-yl nonanoate,
J. 4-( 4-bromophenyI}-1-[4-( 4-fluorophenyl)-4-oxobutyl]

piperidin-4-yl undecanoare,
F'('laoB'
D. 4-( 4-bromophenyI}-I-[4-[4-[4-(4-bromophenyl)-4-
hydroxypiperidin-I-yl] phenylj-4-oxobutyl]piperidin-4-yl
decanoate,
B' ~N I
a
H3C~O·
o
K. 4-(4-bromophenyl)-I-[4-(4-fluorophenyl)-4-oxobutyl]
piperidin-4-yl dodecanoate,
E. 4-(4'-bromobiphenyl-4-yl)-I-[4-(4-fluorophenyl)-4-
oxobutyl]piperidin-4-yl decanoare,
L. 1-(4-fluorophenyl)ethanone.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'hE"
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1-348 Brompheniramine Maleate 2022
E. To 0.15 g in a porcelain crucible add 0.5 g of anhydrous

Brompheniramine Maleate sodium carbonate R. Heat over an open flame for 10 min.
(Ph. HUT. monograph 0977) Allow to cool. Take up the residue in 10 mL of dilute nitric
acidR and filter. To I mL of the filtrate add I mL of
water R. The solutiongives reaction (a) of bromides (2.3.1).
C0 2.H F. Optical rotation (see Tests).
andenanliomer • ( TESTS
Co,H Appearance of solution
than reference solution BY6 (2.2.2, IWethod If).
C,oH"BrN,O, 435.3 980-71-2 Dissolve 2.0 g in methanol R and dilute to 20 mL with the
same solvent.
Action and use pH (2.2.3)
Histamine H I receptor antagonist; antihistamine. 4.0 to 5.0.
PhE" ~ _ Dissolve 0.20 g in 20 mL of carbon dioxide-free waw R.
DEFINITION Optical rotation (2.2.7)
(3R.s)-3-(4-Bromophenyl)-N,N-dimethyl-3-(pyridin-2-yl) -0.20° to + 0.20" (measured in a 2 dm tube).
propan-l-amine (Z)-bulenedioate. Dissolve 2.5 g in water R and dilute to 25.0 mL with the
same solvent.
Content
98.0 per cent to 101.0 per cent (dried substance). Related substances
CHARACTERS
Test soluti<m Dissolve 0.100 g of the substance to be
Appearance
examined in 10.0 mL of methylene chloride R.
Reference solUli<m (a) Dilute 1.0 mL of the test solution to
Solubility
100.0 mL with methylene chloride R. Dilute 1.0 mL of this
Soluble in water, freely soluble in ethanol (96 per cent), in
solution to 10.0 mL with melhylene chloride R.
methanol and in methylene chloride.
Reference solution (b) Dissolve 10 mg of chlorphenamine
IDENTIFICATION maleate CRS (impurity A) and 10 mg of pheniramine
First identification: C~ F. . maleate CRS (impurity C) in methylene chloride R and dilute
Second identification: AJ B, D J E, F. to 5 mL with the same solvent. To 2.5 mL of the solution
A. Melting point (2.2.14): 130"C to 135 "C. add 2.5 mL of the test solution.
B. Ultraviolet and visible absorption spectrophotometry Column:
(2.2.25). - material: fused silica;
- size: 1= 30 rn, 0 = 0.32 mm;
Ten solution Dissolve 65 mg in a 10.3 gIL solution of
- stationary phase: phenyl(50)methyl(50)poIysiioxane R (film
hydrochloric add R and dilute to 100.0 mL with the same
thickness 0.5 pm).
solution. Dilute 5.0 mL of this solution to 100.0 mL with a
10.3 gIL solution of hydrochloric acidR. Oorrier gas nitrogen for chromatography R.
Spectral range 220-320 nm. Flow rat< 1.0 mllmin.
Absorption maximum At 265 nm. Split ratio 1:5.
Specific absorbance at the absorption maximum 190 to 210. Temperature:
C. Infrared absorption spectrophotometry (2.2.24). - column: 205 "Cr
- injulUm port and detector: 250 °C.
Comparison brompheniramine maleate CRS.
Deuaion Flame ionisation.
Injection I ~L.
Run time 1.;2 times the retention time of brompheniramine.
solvent. Identification of impun'ties Use the chromatogram obtained
Reference solution Dissolve 56 mg of maleic acid R in with reference solution (b) to identify the peaks due to
methanol R and dilute to 10 mL With the same solvent. impurities A and C.
Plat< TLC silica gel F m plat< R.' Relative reiemum With reference to brompheniramine
(retention time =about 34 min): impurity C = about 0.4;
Mobile phase water R, anhydrous formic addR, methanol R, impurity A = about 0.7.
di-isopropyl ether R (3:7:20:70 V!V!VIV).
Application 5!!L. - resolution: minimum5.0 between the peaks due to
Development Over 2/3 of the plate. impurity A and brornpheniramine.
Drying In a current of air for 5 min. Limits:
Detection Examine in ultraviolet light at 254 nm. - impurities A, C: for each impurity, not more than 4 times
Results The chromatogram obtained with the test solution me area of the principal peakin the chromatogram
shows 2 clearly separated spots; the upper spot is similar in obtained withreference solution (a) (0.4 per cent);
position and size to the spot-in the chromatogram obtained - unspecified imputities: for each impurity, not more thanthe
with the reference solution. area of the principal peakin the chromatogram obtained
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2022 Bronopol 1-349
- total; not more than 10 times the area of me principal IDENTIFICATION

peakin the chromatogram obtained with reference A. The infrared absorption spectrum, Appendix II A, is
solution (a) (1.0 per cent); concordant with the reference spectrum ofbronopol (RS 031).
~ disregard limit: 0.5 times the area of the principal peak in B. Dissolve 0.1 gin 10 mL of water, add 10 mL of
the chromatogram obtained with reference solution (a) 7.5M sodium hydroxide and, carefully with constant stirring
(0.05 per cent). and cooling, 0.5 g of nkkel-aluminium alloy. Allow the
Loss on drying (2.2.32) reaction to subside, filter and carefully neutralise with nitric
Maximum 0.5 per cent, determined on 1.000 g by drying in add. The resulting solution yields reaction A characteristic of
an oven at 105°C for 3 h. bromides, Appendix VI.
Sulfated ash (2.4.14) C. Melting point, afterdrying over phosplwrus pentoxide at a
Maximum 0.1 per cent, determined on 1.0 g. pressure not exceeding 0.7 kPa, about 130°, AppendixV A.
ASSAY TESTS
Dissolve 0.260 g in 50 mL of anhydrous acetic acid R. Titrate Acidlty or alkalinity
with 0.1 ,"l perchknic acid, determining the end-point pH of a 1% w/v solution, 5.0 to 7.0, AppendixV L.
potentiometrically (2.2.20). . Related substances
1 mL of 0.1 M perchlori< add is equivalent to 21.77 mg Carry out the method for liquid chromatography,
of C2oH23BrN204' Appendix In D, using the following solutions in the.mobile
phase.
STORAGE
Protected from light. (1) 0.2% wlv of the substance being examined.
(2) Dilute a volume of solution (I) to produce a solution
IMPURITIES
containing 0.0002% w/v of the substance being examined.
Specified impurities A, C.
(3) 0.001% wlv each of 2-merhy/-2-nitropropan-l,3-dioland
rris(hydroxymerhyl)nitromethaue.
C
CI
.o':
. I
NH
N ....CH3
i:Ha
and enanUomer
(4) 0.0002% wlv each of 2-methyl-2-nitropropane-I,3-diol,
2-nitroethanol, sodium bromide and tris(hydroxymethyl)
nitromethaneand 0.2% wlv of the substance being
examined.
A. (3RS)-3-(4-chlorophenyl)-N,N-dimethyl-3-(pyridin-2-yl) (a) Use a stainless steel column (15 em x 4.6 mm) packed
propan-I-amine (chlorphenamine), with ocrade<ylsily/ silica gel for chromatography (5 um)
(Phenomenex Luna CIS (2) is suitable).
CN . H
(b) Use isocratic elution and the mobile phase described
below .
~
' CH andenantiomer (c) Use a flow rate of 1 mL per minute.
, N"- 3
I ' (d) Use a column temperature of 35°.

~ CH3
(e) Use a detection wavelength of214 om.
(I) Inject 20 ~L of each solution.
C. (3RS)-N,N-dimethyl-3-phenyl-3-(pyridin-2-yl)propan-l-
amine (pheniramine). (g) For solution (I) allow the chromatography to proceed for
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""E<I at least 3 times the retention time of the principal peak.
MOBILE PHASE
1 volume of a 10% vlv solution of onhophosphoric acid,
10 volumes of aUlOTIitn1e and 189 volumes of water, adjust
Bronopol the pH to 3.0 using 2M sodium hydroxide.
SYSTBM SUITABILITY
Br NO, The test is not valid unless, in the chromatogram obtained
HO~OH with solution (4):
the resolution factor between the peaks due to sodium bromide
200.0 52-51-7 and tris(hydroxymethyl)nitromethane is at least 1.0;
C,HJ!rNO,
the resolution factor between the peaks due to tris
Action and use (hydroxymethyl)nitromethane and 2-nitroethanol is
Antibacterial preservative. at least 1.5.
LI.I\UTS
DEFINITION
Bronopol is 2-bromo-2-nitropropane-1,3-diol. It contains not In the chromatogram obtained with solution (I):
less than 99.0% and not more than 101.0% of C:3H6BrN04, the area of any peak corresponding to 2-methyl-2-
calculated with reference to the anhydrous substance. nitropropane-I,3-diol and tris(hydroxymethyl)nitromethane
are not greaterthan the area of the corresponding peaks in
CHARACTERISTICS the chromatogram obtained with solution (3) (0.5% of each);
White or almost white crystals or crystalline powder.
the area of any other secondary peak is not greater than the
Freely soluble in water and in erhanol (96%); slightly soluble
area of the principal peak in the chromatogram obtained with
in glycerol and in liquid paraffin. solution (2) (0.1 %).
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1-350 Brotizolam 2022
Sulfated ash Test solution Dissolve 50.0 mg of the substance to be

Not more than 0.1 %, Appendix IX A. examined in acetonitrile R and dilute to 50.0 mL with the
Water same solvent.
Not more than 0.5% wlw, Appendix IX C, Method I B. Reference solution (a) Dilute 1.0 mL of the test solution to
Use 5 g. 100.0 mL of acetonitrile R. Dilute 1.0 mL of this solution to
10.0 mL with acetonitrile R.
ASSAY
In a flask fitted with a reflux condenser dissolve 0.4 g in Reference sdution (b) Dissolve 5 mg of me substance to be
15 mL of water and add 15 mL of 7.5M sodium hydroxide. examined and 5 109 of brotizolam impurity B CRS in 50 mL
Slowly,with caution, add 2 g of nickel-aluminium aUoy of auumim·k R. Dilute 2 mL of this solution to 20 mL with
through the reflux condenser, agitating the flask whilst acetonitrile R.
cooling under running water. Allow the mixture to stand for Column:
10 minutes and boil for 1 hour. Cool and filter under - size: 1= 0.15 m, " = 4.6 mm;
reduced pressure, washing the condenser) flask and residue - stationary phase: occy/silyl silica gelfor chromatography R
with 150 mL of water. Combine the filtrate and washings, (5 um);
add 25 mL of nitric acid and 40 mL of O.1M silver nitrate VS, - temperature: 40 °C.
shake vigorously and titrate with O.1M ammonium thiocyanate Mobile phase:
VS using ammonium iron(JIl) sulfate solution R2 as indicator. - mobile phaseA: 2 gIL solutionof sodium heptanesulfonace
Repeat the operation without the substance being examined. monohydrate Rj
The difference between the titrations represents the amount - mobile phase B: mix 25 volumesof a 2 WL solutionof
of silvernitrate required. Each mL of O.1M silvernitrate VS is sodium heptanesulfonau Rand 75 volumesof aceumitri/e Rj
equivalent to 20.00 mg of C3H6 BrNO...
STORAGE (min) (per cent VIJI) (per cent VIJI)
Bronopol should be protected from light.
0-' 63 37
4 - 15 63 --> 12 37 --> 88
Flow rate 2.0 rnUmin.

Brotizolam Deuaion Spectrophotometer at 242 run.
(Ph. Bur. monograph 2197) Injection 5 pL.
Relative retention With reference to brotizolam (retention
H3C
yN'N time = about 7.4 min): impurity A = about 0.5j
=
B'.~S
I ~J
System suitabilily Reference solution (b):
impurity B and brotizolam.
"" CI Limits:
-" - impurity B: not more than the area of the principal peakin
(0.1 per cent);
C 15H IOBrClN,S 393.7 57801-81-7
- unspecified impuniieJ: for each impurity, not more than the
Action and use area of the principal peakin the chromatogram obtained
Benzodiazepine. with reference solution (a) (0.10 pet cent);
PflEu _ the chromatogram obtained with reference solution (a)
(0.2 per cent);
DEFINITION
2-Bromo-4-(2-chlorophenyl)-9-methyl-6H-thieno-[3,2-j)
[1,2,4]-triazolo[4,3-a] [1,4]diazepine.
(0.05 per cent).
Content
Chlorides (2.4.4)
Maximum 100 ppm.
CHARACTERS Dissolve 0.67 g in 20.0 mL of m~hanol R, mix and filter.
Appearance
White or yellowish powder.
Solubility an oven at 105 "C.
Practically insoluble in water, sparingly soluble or slightly
soluble in methanol, slightly soluble in ethanol (96 per cent).
IDENTIFICATION
ASSAY
Dissolve 0.150 g in a mixture of 25 mL of glac£al acetic
Comparison brotizolam CRS. acid Rand 50 mL of acetk anhydride R. Titrate to the second
TESTS point of inflexion with 0.1 j\1 perchlOTic acid, determining the
Related substances end-point potentiometrically (2.2.20).
Liquid chromatography (2.2.29). Cany out the USt protected I mL of 0.1 M perchloric acidis equivalent to 19.68 109
from light aud prepare the solutions immediately before use. of C1sHIOBrClN,S.
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2022 Buclizine Hydrochloride 1-351
IMPURITffiS IDENTIFICATION
Specified impurities B. A. The infrared absorption spearum, Appendix II A, is
Otherdetectable impurities (the following sub,ranm would, if concordant with the reference spectrum of bucHzine
present at a suffidentleveJ, be detected by oneor other of the tests hydrochloride (RS 032).
in the monograph. They arelimit&! by thegeneral acceptance B. A 0.25% w/v solution in ethanol (50%) yields reaction A
criterion for other/unspecified impurities and/or by thegeneral characteristic of chlorides, Appendix VI.
monograph Sub,ranm for phannaceutical use (2034). It is
TESTS
therefore not neussary to identify these ;mpun"ties for
Related substances
demonstration of compliance. Seealso 5.10. Control of impurities
Carry out the method for liquidchromatogrnphy,
in subseanas for pharmaceutical use) A.
Appendix ill D, usingfour solutions in the initial mobile
phase containing (I) 0.0010% wlv of the substance being
examined, (2) 0.50% w/v of the substance being examined,
(3) 0.0010% wlv of 1,4-bis(4-chlorobenzhydryl)
piperazine BPCRS and (4) 0.50% wlv of budizine hydrochloride
impun'ty standard BPCRS.
The chromatographic procedure may be carried out using a
stainless steel column (20 cm x 4 mm) packed with
o<radecylsily/ silica gelfor chromatogrnphy (I 0 urn) (Nudeosil
CIS is suitable). Use as the initial mobile phase O.OIM sodium
A. 4-(2-chlorophenyl)-9-methyl-6H-thieno[3,21l heplanesulfonau in a mixture of 55 volumes of water and
[1,2,4] triazolo [4,3-a] [I ,4]diazepine (desbromobrotizolam), 45 volumes of cueten;tr'-k and as the final mobile phase O.OIM
sodium heptanesulfonate in a mixture of 20 volumes of water
and 80 volumesof acetonitrile. Before use, adjust the pH of
both the initial and final mobile phases to 4.0 with 1M
orchophosphom acid. Carry out a linear gradient elutionwith a
flow rate of 2 mL per minute for 30 minutes and maintain
the final mobilephase for 10 minutes with the same flow
rate. Use a detectionwavelength of 230 om.
The test is not valid unless the chromatogram obtained with
solution (4) closelyresembles the chromatogram supplied
with buelizine hydrochloride impuri,ystandard BPCRS.
B. 2-bromo-4-(2-chlorophenyl)-6H-thieno[3,2-A In the chromatogram obtained with solution (2) the area of
[1,2,4]triazolo[4,3-a] [1,4]diazepine any peak corresponding to 1,4-bis(4-chlorobenzhydcyl)-
(desmethylbrotizolam). piperazine is not greater than the area of the peakobtained in
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" the chromatogram with solution (3) and the area of any
other secondary peak is not greater than the area of the peak
in the chromatogram obtained with solution (I).
Loss on drying
Buclizine Hydrochloride When dried to constant weight at 1000 to 105Cl, loses not
more than 1.0% of its weight. Use I g.
Cl Sulfuted ash
Not more than 0.1%, Appendix IX A.
ASSAY
H
Carry out Method I for non-aqueous titradon,
,2HCI
Appendix VIII AJ using 0.4 g and determining the end point
potentiometrically. Each mL of 0.1M perchlori< acid VS is
equivalent to 25.30 mg of C28 H 33 ClN 2,2HCI.
CuH"CIN,,2HCI 506.0 129-74-8 IMPURITffiS
A. I ,4-bis(4-chlorobenzhydcyl)piperazine,
Action and use B. 4-chlorobenzhydrol, 1-(4-chlorobenzhydcyl)piperazine,
Histamine Hi receptor antagonist; antiemetic.
4-chlorobenzophenone.
DEFINITION
Buc1izine Hydrochloride is (RS)-I-( 4-tert-butylbenzyl)-4-(4-
chlorobenzhydryl)piperazine dihydrochloride. It contains not
less than 99.0% and not more than 100.5% of
C2sH33ClN2,2HCI, calculated with reference to the dried
substance.
CHARACTERISTICS
A white or slightly yellowish, crystalline powder.
Practically insoluble in water; sparingly soluble in propane-l,2-
dial; very slightly soluble in ethanol (96%).
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1-352 Budesonide 2022
Results A The principal spot in the chromatogram obtained

Budesonide with the test solution is similar in position andsize to the
(Ph. Hur. monograph 1075) principal spot in the chromatogram obtained with reference
solution (a).
OH Detection B Spray with alcoholic solution of sulfuric acid R;
heat at 120 "C for 10 min or until the spots appear and allow
... ,_/~,C,_H3""07< <:: CH3 to cool; examine the chromatograms in daylight and in
.. 0 H and epimer at C' ultraviolet light at 365 nm.
H Results B The principal spot in the chromatogram obtained
with the test solution is similar in position, colour in daylight,
o fluorescence in ultraviolet lightat 365 nm and size to the
430.5 51333-22-3 solution (a).
Action and use System suitability Reference solution (b):
Glucocorticoid. - the chromatogram shows 2 clearly separated spots.
C. Dissolve about 2 mg in 2 mL of sul/um add R. Within
Preparations
5 min a yellow colourdevelops. Within 30 min the colour
Budesonide Aqueous Nasal Spray
changes to brown or reddish-brown. Cautiously add the
Budesonide Inhalation Powder solution to 10 mL of water R and mix. The colour fades and
Budesonide Inhalation Powder, pre-metered a clear solutionremains.
Budesonide Nebuliser Suspension D. Dissolveabout J mg in 2 mL of a solution containing 2 g
Budesonide Pressurised Inhalation of phosphomolybdic acid R dissolved in a mixture of 10 mL of
dilutesodium hydroxide solution R, 15 mL of water Rand
PhE" _
25 mL of glacial acetic acid R. Heat for 5 min on a water-
DEFINlTION bath. Cool in iced water for 10 min and add 3 mL of dl7ute
Mixture of the C-22S (epimer A) and the C-22R (epimer B) sodium hydroxide solution R. The solution is blue.
epimers of l6cx,17-[(IRS}-butylidenebis(oxy)]-llp,21- TESTS
d jhydroxypregna- 1,4-diene-3,20-dione. Related substances
Content liquid chromatography (2.2.29). Carry oUl the testprotected
97.5 per cent to 102.0 pen cent (dried substance). from ligh,.
CHARACTERS Solvent mixture acetonitrile R, phosphate buffer solution
Appearance pH 3.2 R (32:68 VIV).
White or almost white, crystalline powder. Test solution (aJ Dissolve 50 mg of the substance to be
examined in 15 mL of acetonim1e R anddilute to 50 mL with
Solubility
Practically insoluble in water, freely soluble in methylene phosphare buffersolution pH 3.2 R.
chloride, sparingly soluble in ethanol (96 per cent). Test solution (b) Dissolve 25.0 mg of the substance to be
examined in 15 mL of acetonitrile R and dilute to 50.0 mL
IDENTIFICATION with phosphore buffersolution pH 3.2 R.
Firs' identijicatWn: A. Reference solutWn (a) Dilute 1.0 mL of test solution (a) to
Second identification: B" C, D. 10.0 mL with the solvent mixture. Dilute 1.0 mL of this
A. Infrared absorption spectrophotometry (2.2.24). solution to 100.0 mL with the solvent mixture.
Comparison budesonide CRS. Reference solution (b) Dissolve 5 mg of budesonide for system
B. Thin-layer chromatography (2.2.27). suiiabilily CRS (containing impurities A, D, G, K and L) in
Solven'mixture methanol R, methylene chloride R (10:90 VIV). 1.5 mL of acetonitrile R and dilute to 5 mL with phosphore
buffersolution pH 3.2 R.
examined in the solvent mixture and dilute to 10 mL with Reference solution (c) Dissolve 25.0 mg of budesonide CRS in
the solvent mixture. 15 mL of acetonitrile R and dilute to 50.0 mL with phosphore
buffer solution pH 3.2 R.
Reference sdution (a) Dissolve 25 mg of budesonide CRS in
the solvent mixture and dilute to 10 mL with the solvent Column:
mixture. - size: 1= 0.15 m, 0 = 4.6 mm;
- stationary phase: end-capped octaduylsilyl silica gelfor
Reference solutWn (b) Dissolve 12.5 mg of triamcinolone
acetonick CRS in reference solution (a) and dilute to 5 mL
with reference solution (a).
Mobile phase:
ne« TLC silica gel Fm plare R. - mobile phase A: anhydrous ethanol R, acetonitrile R,
Mobile phase Add a mixture of 1.2 volumes of water Rand phosphore buffersolution pH 3.2 R (2:32:68 VIVIV);
8 volumes of methanol R to a mixture of 15 volumes of - mobile phase B: acetonitrile R, phosphore buffer solutWn
ether Rand 77 volumes of methylene chloride R. pH 3.2 R (50:50 VIV);
Applica'ion 5 pL.
Drying In air.
Detection A Examine in ultraviolet light at 254 om.
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2022 Budesonide 1-353
Tim. Mobile phase A Mobile phase B Time Mobile phase A MobUe phase B
(min) (per cent lim (per cent Vm (min) (per cent VIJ') (per cent VIJI)
0-38 100 o 0-21 tOO o
38 - 50 100 ..... 0 0---1 100 21 - 22 100 ..... 0 0-+ 100
50 - 60 o 100 22 - 31 o tOO
Flow rale 1 mUmin. Injection 20 J.lL of test solution (b) and reference
Detection Spectrophotometer at 240 om. solutions (b) and (c).
Injection 20 J.1L of test solution (a) and reference =
Retention time Budesonide epimer B about 17 min;
solutions (a) and (b). budesonide epimer A ;;:; about 19 min.
Identification of impuruies Use the chromatogram supplied SYSl<m suitabi/i/JI:
with budesonide for system suitabiiliy GRS and the - resolution: minimum 1.5 between the 2 principal peaks
chromatogram obtained with reference solution (b) to (budesonide epimers A and B) in me chromatogram
identify the peaks due to impurities A, D, G J K and L. obtained with reference solution (c);
Relativeretention With reference to budesonide epimer B - peak-to-vaJ/ey ratio: minimum 3, where Hp == height above
the baseline of the peak due to impurity Land
(retention time ;;;; about 17 min): impurity A = about 0.1;
epimers of impurity D = about 0.63 and 0.67;
H; == height above the baseline of the lowest point of the
=
impurity L about 0.95; epimers of impurity G = about 1.2
budesonide epimer B (the I S[ of the 2 principal peaks) in
and 1.3; epimers of impurity K = about 2.9 and 3.0.
the chromatogram obtained with reference solution (b).
System miMbility Reference solution (b):
- peak-to-fJaney ratio: minimum 2.5, where Hp = height Limit:
above the baseline of the 1Sf of the 2 peaks due to
- epimer A: 40.0 per cent to 51.0 per cent of the sum of the
areas of the 2 peaks due to the budesonide epimers.
impurity G and H; == height above the baseline of the
lowest point of the curve separating this peak from the Loss on drying (2.2.32)
peak due to budesonide epimer A (the 2 nd of the Maximum 0.5 per cent, determlned on 1.000 g by drying in
2 principal peaks); and minimum 3, where H p == height an oven at 105 "C.
above the baseline of the peak due to impurity Land ASSAY
H; = height above the baseline of the lowest point of the Liquid chromatography (2.2.29). Examine the
curve separating this peak from the peak due to chromatograms obtained in the test for epimer A.
budesonide epimer B (the 1" of the 2 principal peaks).
Calculate the percentage content of C25H34.06 from the sum
Limits: of the areas of the 2 peaks due to the budesonide epimers
- correction factors: for the calculation of content, multiply and the declared content of budesonide CRS.
=
corresponding correction factor: impurity D 1.8; IMPURITIES
impurity K = 1.3; Specified impurilies AJ DJ s; L.
- impurities AJ L: for each impurity, not more than twice the Other dete<tabk impurities (thefollowing substances would, if
sum of the areas of the 2 peaks due to the budesonide present at a sufficientlevd, be detected by oneor otherof the tests
epimers in the chromatogram obtained with reference in the monograph. They arelimited by thegeneral acceptance
solution (a) (0.2 per cent); criterion for other/unspecified impunues and/or by the general
- impurities D, K: for each impurity, for the sum of the monograph Substanoofor pharmaceutical use (2034). It is
areas of the 2 epimer peaks, not more than twice the sum therefore not necessary to identify these impurities for
of the areas of the 2 peaks due to the budesonide epimers demonstration of compliance. See also 5.10. Control of impurities
in the chromatogram obtained with reference solution (a) in subseanus for pharmaceutical use) BJ C, EJ F J G, H, I, J.
(0.2 per cent);
- unspecified impurities: for each individual peak, not more
than the sum of the areas of the 2 peaks due to the
budesonide epimers in the chromatogram obtained with
- total: not more than 5 times the sum of the areas of the
2 peaks due to the budesonide epimers in the
chromatogram obtained with reference solution (a) o
(0.5 per cent);
- disregard limit, 0.5 times the sum of the areas of the A. 11~, 16., 17,21-tetrahydroxypregna-1,4-diene-3,20-dione,
2 peaks due to the budesonide epimers in the
(0.05 per cent).
EpimerA and epimer at C'
JWobi/e phase: o
B. 16a,17-[(lRS)-ethylidenebis(oxy)1-11~,21
dihydroxypregna-l,4-diene-3,20-dioneJ
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1-354 Bufexamac 2022
o OH
.. OX~CH,
"0 H
H andeplmeralC'
o
o
C. 100,17-[(IRS)-butylidenebis(oxy)]-11 ~-hydroxy-17
(hydroxymethyl)-D-homoandrosta-l ,4-diene-3,I 'ra-dione, I. 11~,17,21-nihydroxy-3,20-dioxopregna-1,4-dien-loo-yl
butanoate,
o
CHO OH
...07<~CH,
'.0 H and eplmeral C' .. O;:;;~CH,
H --0 H and eplmeral C'
H
o
o
D. 100,17-[(IRS)-butylidenebis(oxy))-II )l-hydroxy-3,20-
dioxopregna-I ,4-dien-21-al, J. 100,17-[(IRS)-butylidenebis(oxy) 1-9~-bromo-11 ~,21
dihydroxypregna-l,4-diene-3,ZO-dione,
OH
o
o
CH3 ._0
'~'./'-,I --<
.................. c~
..~H and eplmel8lC'
oJ....
CH,
H "..o--;.;:~CH,
-...0 H and eplmer at C'
o H
E. 100, 17-[(IRS)-butylidenebis(oxy))-11 ~,21 o

dihydroxypregna-I,4, 14-triene-3,2Q-dione,
OH
K. 100,17-[(IRS)-butylidenebis(oxy))-11 ~,21
o dihydroxypregna-l,4-diene-3,20-dione-21-acetate,
F. 100,17-[I-methylethylidenebis(oxy))-II ~,21 o
dihydroxypregna-l,4-diene-3,ZO-dione,
L. 100,17-[( lRS)-butylidenebis(oxy))-21-hydroxypregna-I,4-
OH
o diene-3,II,20-trione.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE,;
CH3 .. 0 ••. -..........,...CH3
',,"./',1--' ..~H andeplmeralC'
Bufexamac
G. 1OO,17-[(IRS)-butylidenebis(oxy)]-11 ~,21 (ph. Eur. monograph 1179)
dihydroxypregn-4-ene-3,20-dione.
OH
o
CH3 ._0 * ................. CH3
/ ' , 1 -.... __~H andepimeralC'
H 2438-72-4
o Action and use

Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.
H. 16~, 17-[(IRS)-butylidenebis(oxy)]-21-hydroxypregnal,4,9
PhE,; _
(ll)-niene-3,ZO-dione,
DEFINITION
2_(4-Butoxyphenyl)-N-hydroxyace,amide.
Content
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2022 Bufexamac 1-355
CHARACTERS Mobile phase:

Appearance - mobile phase A: mix 30 volumes of a 1.4 gIL solution of
White or almost white, crystalline powder. dipotassium hydrogen phosphare Rand 70 volumes of
Solubility methanol RJ then adjust to pH 3.6 with dilute phosphoric
Practically insoluble in water,soluble in dimethylfonnamide, acid Rj
slightly soluble in ethyl acetate and in methanol. - mobile phase B: methanol R;
IDENTIFICATION Time Mobile phase A Mobile phase B

First identification: B. (min) (per cent V/l? (per cent V/l?
Second identification: A, C. 0·10 100 o
10 - 13 100 ..... 70 0 ..... 30
A. Ultravioletand visible absorption spectrophotometry
(2.2.25). 13 - 40 70 30
Test solution Dissolve 20 mg in methanol R and dilute to

20 mL with the same solvent. Dilute 1 mL of the solution to Frow rare I mUmin.
50 mL with methanol R. Detection Spectrophotometer at 275 run.
Spectral range 210-360 run. Injection 20 ~L.
Absorption maxima At 228 nm, 277 run and 284 nm. Identification of impuniies Use the chromatogram obtained
B. Infrared absorption spectrophotometry (2.2.24). with referencesolution (b) to identify the peaksdue to
impurities A, B, C and D.
Comparison bofexamac CRS.
Relative retention With reference to bufexamac (retention
time = about 5.7 min): impurity D = about 1.3;
Test solution Dissolve 10 rng of the substance to be impurity A = about 1.8,; impurity B = about 3.0;
examined in methanol R and dilute to 5 mL with the same impurity C = about 5.4.
solvent.
System suitability Referencesolution (b):
Reference solution (a) Dissolve 20 mg of bufexamac CRS in - resolution: minimum 2.0 between the peaks due to
methanol R and dilute to 10 mL with me same solvent. bufexamac and impurity D.
Reference sdution (b) Dissolve 10 mg of salicylic acid R in Limits:
reference solution (a) and dilute to 5 rnL with the same - impurities A, B, C, D: for each impurity) not more than
solution. the area of the principal peak in the chromatogram
Plate TLC silica gel F2 , . piau R. obtained with referencesolution (a) (0.2 per cent);
Mobile phase glacial acetic acidR, dioxan R, toluene R - unspecified impurities: for each impurity, not more than
(4:20:90 VIVIV). 0.5 times the area of the principal peak in the
Applicaewn 10 ~L. chromatogram obtained with reference solution (a)
(0.10 per cent);
Drying In a current of wann air. peak in the chromatogram obtained with reference
Detection Examine in ultraviolet light at 254 run. solution (a) (0.5 per cent);
System suitability Reference solution (b): - disregard limit: 0.25 times the area of the principal peak in
- the chromatogram shows 2 clearly separated spots. the chromatogram obtained with reference solution (a)
Results The principal spot in the chromatogram obtained (0.05 per cent).
with the test solution is similar in position and size to the Loss on drying (2.2.32)
principal spot in the chromatogram obtained with reference Maximum 0.5 per cent, determined on 1.000 g by drying in
solution (a). vacuo at 80 "C for 3 h.
TESTS Sulfated ash (2.4.14)
uqwdchromarography(~~2~. ASSAY
Test solution Dissolve 50.0 mg of the substance to be Dissolve 0.200 g in 50 mL of dimeJhylformamide R. Titrate
examined in mobile phase A and dilute to 20.0 mL with with 0.1 lvt lithium methoxide, determining the end-point
mobile phase A. potentiometrically (2.2.20).
Reference solution (a)Dilute 5.0 mL of the test solution to 1 mL of 0.1 M lithium methoxide is equivalent to 22.33 mg of
25.0 mL with mobile phase A. Dilute 1.0 mL of dtis solution C12H17N03'
STORAGE
Reference sonaion (b) Dissolve 5 mg of bofexomac CRS, Protected from light.
5 mg of bufexamac impurity A CRS, 5 mg of bofexamac
impurity B CRS, 5 mg of bufexamac impurity C CRS and IMPURITIES
5 mg of bufexamac impurity D CRS in mobile phase A and Specified impurities A~ B~ C, D.
dilute to 10.0 mL with mobile phase A. Dilute 1.0 mL of the
Column:
- size: 1:;;;: 0.25 m, 0 = 4.6 nun;
- stationary phase: end-capped oaade<ylsi/yl silica gelfor
A. 2-(4-butoxyphenyl)acetic acid,
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1-356 Buflomedil Hydrochloride 2022

solvent.
Reference solution Dissolve 40 mg of bufiomedil
B. methyI2-(4-butoxyphenyl)acetate, hydrochloride CRS in methanolR and dilute to 2 mL with the
same solvent.
PIal< TLC silica gel Fm plate R.
Mobile phase triethylamine R) 2-propanol RJ wluene R
(5:50:50 VIVIV).
C. butyl 2-(4-butoxyphenyl)acetate, Applicmion 10 ut,
Drying In air.
D.2-(4-butoxyphenyl)acetamide. with the test solutionis similar in position and size to the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEur principal spot in the chromatogram obtained with the
reference solution.
TESTS
Buflomedil Hydrochloride Solution S
(ph. Bur. monograph 1398) 50 mL with the same solvent.
Solution S is clear (2.2.1) and colourless (2.2.2, Method 11)•
. Her pH (2.2.3)
Related substances
343.9 35543-24-9 Liquid chromatography (2.2.29).
Action and use examined in the mobilephase and dilute to 10.0 mL with
Vasodilator. the mobile phase.
PhE" _ Reference solution (a) Dilute 0.5 mL of the test solution to
DEFINITION solution to 10.0 mL with the mobile phase.
4-(Pyrrolidin-I-yl)-I-(2,4,6-trimethoxyphenyl)butan-L-one
Reference solutiou (b) Dissolve 2 mg of bujlomedil
hydrochloride.
impuri/jl B CRS in the mobile phase, add 0.5 mL of the test
Content solution and dilute to 100.0 mL with the mobile phase.
98.5 per cent to 101.5 per cent (dried substance). Reference solution (c) Dissolve the contents of a vial of
CHARACTERS bujlomedil for peak identification CRS (containing impurities A
Appearance and C) in 1.0 mL of reference solution (b).
White or almost white, microcrystalline powder. Column:
Solubility - size: 1= 0.25 m, 0 = 4.6 mm;
Freelysoluble in water, soluble in ethanol (96 per cent), very - stationary phase: end-capped octadecylsilyl silica gelfor
slightly soluble in acetone. chromatography R (5 pm);
mp
About 195 °C, with decomposition. Mobile phase Mix 45 volumes of acetonitrile R1 and
55 volumes of a 9.25 gIL solution of potassium dihydrogen
IDENTIFICATION phosphal< R adjusted to pH 2.5 with phospho';'; acid R.
Flow ral< I mUmin.
Second idemificatian: A, C, D.
Injection 10 ~ of the test solution and reference
(2.2.25).
solutions (a) and (c).
Test solution Dissolve 25.0 mg in ethanol (96 per cen!! Rand Run time Twice the retention time of buflomedil.
dilute to 50.0 mL with the same solvent. Dilute 2.0 mL of
the solution to 20.0 mL with ethanol (96 per<en!! R. 1dentijicarWn of impurities Use the chromatogram supplied
with bujlomedil for peak identification CRS and the
Spectral range 220"350 om.
Absorption maximum At 275 om. the peaksdue to impurities A, Band C.
Specific absorbance at the absorption maximum 143 to 149. Relativeretention With reference to buflomedil (retention
B. Infrared absorption spectrophotometry (2.2.24). time = about 5 min): impurity B = about 0.6;
Comparison bujlomedil hydrochloride CRS. impurity C = about 0.7; impurity A = about 1.5.
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2022 Bumetanide 1-357
System suitabIlity Reference solution (c):

Bumetanide
impurity B and impurity C.
Limits:
- impuniies A, B, C: for each impurity, not more man the
chromatogram obtained with reference solution (3)
(0.10 per cent);
- rotal: not more than twice the area of the principal peak in
(0.5 per cent); 364.4 28395-03-1
- disregard limit: 0.2 times the area of the principal peak in Action and use
the chromatogram obtained with reference solution (a) Loop diuretic.
(0.05 per cent).
Preparations
Los. on drying (2.2.32) Bumetanide Injection
Bumetanide Oral Solution
Bumetanide and PotassiumProlonged-release Tablets
Maximum 0.1 per cent, determined on 1.0 g. Bumetanide Tablets
PhEIr _
ASSAY
Dissolve 0.300 gin 15 mL of anhydrous aceti< acid R and add DEFINITION
35 mL of acetic anhydride R. Titrate with 0.1 M perchloric 3-(Butylamino}-4-phenoxy-5-sulfamoylbenzoic acid.
acid, determining the end-point potentiometrically (2.2. 2{}).
Content
I mL of 0.1 M perchlonc acid is equivalent to 34.39 mg of
C17H26ClN04'
CHARACfERS
IMPURITIES
Appearance
Specified impuriti<s A, B, C. White or almost white, crystalline powder.
OCH 30 0 Solubility
~N
Practically insoluble in water, soluble in acetone and ln.
ethanol (96 per cent), slightly soluble in methylene chloride.
H,eo "" I OH It dissolves in dilute solutions of alkali hydroxides.
A. 1-(2-hydroxy-4,6-dimethoxyphenyl}-4-(pyrrolidin-I-yl} mp
buren-Lone, About 233 "C.
~N
0
OCH30
IDENTIFICATION
Comparison bumeumide CRS.
HO~OCH3 If the spectra obtained in the solid state show differences,
dissolve the substance to be examinedand the reference
B. 1-(4-hydroxy-2,6-dimethoxyphenyl}-4-(pyrrolidin-I-yl} substance separately in acetone R, evaporate to dryness and
butan-I-one, record new spectra using the residues.
TESTS
Method11).
Dissolve 0.1 g In a 6 gIL solution of potassium hydroxide R
C. 1-(2,4-dihydroxy-6-methoxyphenyl}-4-(pyrrolidin-I-yl} and dilute to 20 mL with the same solution.
butan-j-one. Related substances
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEIr Liquid chromatography (2.2.29).
the mobile phase.
Reference solution (1)) Dissolve 2 mg of bumetanide
impurity A CRS and 2 mg of bumeumide impurity B CRS in
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1-358 Bupivacaine Hydrochloride 2022
phase. Dilute 1.0 mL of this solution to 100.0 mL with the o,\ II0
mobile phase. H,N / S q C o , H I
Column: o ">.
~ size: I == 0.15 m, 0;;;; 4.6 nun,
- stationary phase: end-capped oClylsilyl silica gelfor
chromatography R (3.5 pm).
Mobile phase Mix 70 volumes of methanolR, 25 volumes of
waterfor chromatography Rand 5 volumes of a 27.2 gIL
6
Id'
B. 3-amino-4-phenoxy-5-sulfamoylbenzoic acid,
NH,
solution of potassium dihydrogen phosphate R previously

adjusted to pH 7.0 with a 280 gIL solution of potassium
hydroxide R; add tetrahexylammonium bromide R to this
mixture to obtain a concentration of 2.17 gIL.
FIbw rate 1.0 mUmin.
Injection 10 ~L.
Run time 5 times the retention time of bumetanide.
Relative retention With reference to bumetanide (retention C. butyl 3-(butylamino)-4-phenoxy-5-sulfamoylbenzoate,
time > about 6 min): impurity B == about O.4j
=
impurity A : :; : about 0.6; impurity C about 2.5j o\\ II0
H,N 'S~Co,H
I
- resolution: minimum 2.0 between the peaks due to o ~ H ..r - CH3 and enanUomer
6
impurity A and impurity B. HN ~CH'
Limits: Id'
- impwidesA, B, C, D: for each impurity, not more than
D.3-[[(2RS)-2-ethylhexyl]amino]-4-phenoxy-5-
obtained with reference solution (a) (0.1 per cent),
sulfamoylbenzoic acid.
- any otherimpun'ty: for each impurity, not more than the
___________ ~ PhE"
with reference solution (a) (0.1 per cent),
- total: not more than twice the area of the principal peak In
(0.2 per cent), Bupivacaine Hydrochloride
the chromatogram obtained with reference solution (a) (ph. Eur. monograph 0541)
(0.05 per cent).
Lo ss on drying (2.2.32) CH, (! H.
CxI ~0~"""
Maximum 0.5 per cent, determined on 1.000 g by drying in andenanUomer
an oven at 105 °C for 4 h. 0 I
~ CH3 ~CH3
342.9 73360-54-0
ASSAY
Dissolve 0.300 g in 50 mL of alcohol R. Add 0.1 mL of Acdon and use
phenol red solution R. Titrate with 0.1 M sodium hydroxide Local anaesthetic.
until a violet-red colour is obtained. Carry out a blank Preparadons
titration. Bupivacaine Injection
1 mL of 0.1 M sodium hydroxide is equivalent to 36.44 mg of Bupivacaine Heavy Injection
C 17H , oN, O, S.
Bupivacaine and Adrenaline InjectionIBupivacaine and
STORAGE Epinephrine Injection
Protected from light. Bupivacaine and Diarnorphine Injection
IMPURITIES Bupivacaine and Fentanyl Injection
Specified impurities A, B, C, D. PhE" _
DEFINITION
(2RS)-I-Butyl-N-(2,6-dimethylphenyl)piperidine-2-
carboxamide hydrochloride monohydrate.
Content
CHARACTERS
Appearance
A. 3-nitro-4-phenoxy-5-sulfamoylbenzoic acid, White or almost white, crystalline powder or colourless
crystals.
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2022 Bupivacaine Hydrochloride 1-359
Solubility Reference solution (a) Dissolve 10 mg of the substance to be

Soluble in water, freely soluble in ethanol (96 per cent). examined, 10 mg of bupiuacaine ;mpun"ty B CRS and 10 mg
of bupivacaine ;mpw;ty E CRS in 2.5 mL of water R, add
IDENTIFICATION
2.5 mL of dilute sodium hydroxide solution R and extract with
First identification: A, D, E.
2 quantities) each of 5 mL, of the internal standard solution.
Second identification: BJ C, D, E. Filterthe lowerlayer and dilute to 20 mL with the internal
A. Infrared absorption spectrophotometry (2.2.24). standard solution.
Comparison bupivacaine hydrochloride CRS. Reference solution (b) Dilute 1.0 mL of the test solution to
B. Thin-layer chromatography (2.2.27). 100.0 mL with the internal standard solution.
Testsolution Dissolve 25 mg of the substance to be Reference solution (c) Dilute 5.0 mL of reference solution (b)
examined in methanol R and dilute to 5 mL with the same to 10.0 mL with the internal standard solution.
solvent. Reference solution (d) Dilute 1.0 mL of reference
Reference solution Dissolve 25 mg of bupivacaine solution (b) to 10.0 mL with the internal standard solution.
hydrochloride CRS in methanolR and dilute to 5 mL with the Column:
same solvent. - material: fused silica;
Plate TLC silica gel G plate R. - size: I = 30 01, 0 = 0.32 rom;
Mobile phase ccnetntrated ammonia R, methanol R - stationary phase: phenyl(5)methy/(95)pa/ysilo:cane R (film
(0.1:100 VIV). thickness 0.25 pm),
Application 5 ~L. Carrier gas helium for chromatography R.
Development Over a path of 10 em. Flow rate 2.5 mUmin.
Drying In air. Split ratio 1:12.
Deration Spray with dilute potassium iodobismUlhate Temperature:
solution R.
Time Temperature
Results The principal spot in the chromatogram obtained (mln) CCJ
o 180
the principal spot in me chromatogram obtained with the 0-10 180 --> 230
Column
reference solution. 10 - 15 230
C. Dissolve 0.1 g in 10 mL of water R, add 2 mL of dIlute Injectionport 250
sodium hydroxide sduuon R and shake with 2 quantities, each Detector 250
of 15 mL, of 1,I-dimethylethy/methyl etherR. DIY the
combined upper layers over anhydrous sodium sulfate Rand
filter. Evaporate the filtrate, recrystallise the residue from
ethanol (90 per cent VII-? R and dry under reduced pressure. Injection I ~L.
The crystals melt (2.2.14) at 105°C to 108 °C. Identification of impun"ties Use the chromatogram obtained
D. It gives reaction (a) of chlorides (2.3.1). with reference solution (a) to identify the peaks due to
impurities Band E.
E. Optical rotation (see Tests).
Relative retention With reference to bupivacaine (retention
TESTS time = about 10 min): impurity B = about 0.7;
Solution S impurity E =about 1.1; internal standard =about 1.4.
Dissolve 1.0 g in carbon dioxide-free water R and dilute to System suitability Reference solution (a):
50 mL with the same solvent. - resolution: minimum 3.0 between the peaks due to
Appearance of solution bupivacaine and impurity E.
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). Limits:
Acidity or alkalinity - impun'ty B: calculate the ratio (R j ) of the area of the
To 10 mL of solution S add 0.2 mL of 0.01 M sodium principal peak to the area of the peakdue to the internal
hydroxide; the pH (2.2.3) is not less than 4.7. Add 0.4 mL of standard from the chromatogram obtained with reference
O.OllH hydrochloric acid; the pH is not greater than 4.7. solution (c); from the chromatogram obtained with the
Optical rotation (2.2.7) test solution, calculate the ratio of the area of the peak
-0.10° to + 0.10°. due to impurity B to the area of the peakdue to the
internal standard: thisratio is not greater than R1
Dissolve 1"0 g in methanol R and dilute to 20.0 mL with the
(0.5 per cent);
same solvent.
- unspecified impurities: calculate the ratio (R,) of the area of
Related substances the principal peak to the area of the peakdue to the
Gas chromatography (2.2.28). internal standard from the chromatogram obtainedwith
Internal standardsolution Dissolve 25 mg of methyl reference solution (d); from the chromatogram obtained
behenate R in methylene chloride R and dilute to 500 rnL with with the test solution, calculate foreach impurity the ratio
the same solvent. of the area of any peak, apart from the principal peak, the
Test solution Dissolve 50.0 mg of the substance to be peakdue to impurity B and the peakdue to the internal
examined in 2.5 mL of water R) add 2.5 mL of dl1ute sodium standard) to the area of the peakdue to the internal
hydroxide solution R and extract with 2 quantities, each of standard: this ratio is not greater than Rz (0.10 per cent);
5 mL, of the internal standard solution. Filterthe lower - total: calculate the ratio (R3 ) of the area of the principal
layer. peak to the area of the peakdue to the internal standard
from the chromatogram obtained withreference
solution (b); from the chromatogram obtained with the
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1-360 Bupivacaine Hydrochloride 2022
test solution, calculate the ratio of the sum of the areas of Sulfated ash (2.4.14)
any peaks, apart from the principal peak and the peak due Maximum 0.1 per cent) determined on 1.0 g.
to the internal standard, to the area of the peakdue to the
ASSAY
internal standard: this ratio is not greater than RJ
Dissolve0.250 g in a mixture of 20 mL of water Rand
(1.0 per cent);
25 mL of ethanol (96 per cent) R. Add 5.0 mL of 0.01 M
- disregard li»ni: ratio less than 0.05 times R3
hydrochloric acid. Carry out a potentiometric titration (2.2.20),
(0.05 per cent).
using 0.1 M ethonotic sodium hydroxitk. Read the volume
Impurity F added between the 2 points of inflexion.
Liquid chromatography (2.2.29). Prepare the solutions 1 mL of 0.1 1\11 ethanolic sodium hydroxide is equivalent to
immediately before use. 32.49 mg of ClsH29CIN20.
examined in mobilephase A and dilute to 10.0 mL wil:h STORAGE
mobile phase A. Protected from light.
Reference solution (a) Dissolve 5.0 mg of bupivcu:aine IMPURITIES
impurity F CRS in mobile phase A and dilute to 100.0 mL Specified impurities B F. J
with mobilephase A. Dilute J.O mL of the solution to Otherdetectable impurities (the following substanc.es would if
J
100.0 mL with mobile phase A. Dilute 1.0 mL of this present at a sufficient level) be deuaed by oneor other of the usts
solution to 10.0 mL with mobile phase A. in the monograph. They are limited by thegeneral aueptanu
Reference solution (b) Dissolve 20 mg of metlryl benzoate R criterion for other/unspecified impurities and/or by the general
and 25 mg of bupivcu:aine impurity F CRS in mobile phase A monograph Substances for pharmaceutical use (2034). It is
and dilute to 50.0 mL with mobile phase A. Dilute 3.0 mL therefore not necessary Ie idenuJy these impurities for
of me solution to 50.0 rnL with mobile phase A. Dilute demonstration of compliance. See also5.10. Control of impurities
1.0 mL of this solution to 10.0 mL with mobile phase A. in substances for pharmaceutical use) A J CJ DJ E.
Column:
~~y0
- size: 1 = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped oaade<y/silyl silica gelfor
chromawgraphy R (5 pm).
Mobile phase: Uo CH,
- mobile phase A: dissolve 0.23 g of sodium dihydrogen
phosphate monohydrate R .and 3.626 g of disodium hydrogen A. N-(2,6-<limethylphenyl)pyridine-2-carboxamide,
phosphate dihydrate R in water R and dilute to 1000 mL
-
with the same solvent; mix equal volumes of this solution
(pH 8.0) and acetonitrile R;
mobile phase B: aceumitrile R; h
CH, yQ~
~ H ••
and enanliomer
Tim, Mobile phase A MobUe phase B

Uo CH,
(min) (per cent Vm (per cent Vm
o ~ 10 100 0 B. (2RS)-N-(2,6-<limethylphenyl)piperidine-2-carboxamide,
10 • 15 100 -Jo 80 0 ..... 20
~Q
15·25 80 20

lJlo CH,
Injection 50~.
Identification 0/impurities Use the chromatogram obtained C. 1-(2,6-dimethylphenyl)-1,5,6, 7-tetrahydm-2H-azepin-2-
with reference solution (a) to identify the peak due to one,
impurity F.
Relative retention with reference to bupivacaine (retention
time = about 20 ruin): impurity F = about 0.3; methyl
benzoate = about 0.4.
System suitability:
- resolution: minimum 4.0 between the peaks due to D. (2RS)-2,6-<1ichloro-N-(2,6-<1imethylphenyl)hexanamide,
impurity F and methyl benzoate in the chromatogram
obtained with reference solution (b);
- signal-to-noise ratio: minimum 40 for the principal peakin
Limit:
- impw;t;y F: not more than me area of the principal peak in
E. 6-(butylamino)-N-(2,6-dimethylphenyl)hexanamide,
(10 ppm).
drying in an oven at lOS "C.
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2022 Buprenorphine 1-361
Reference solution (b) Dissolve 5 mg of buprenorphine for

system suitability CRS (containing impurities A, B, F, G, H
and n in 1.0 mL of methanolR.
Column:
F. 2,6-dimedlylaniline.
=
- size: I 0.05 m, 0 = 4.6 mm;
- stationary phose: end-capped octad«y/si/yl silica gelfor
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _~PhE"' chromatography R (3.5 pm),
Mobik phose:
- mobl7e phaseA: mix 10 volumes of acetonitrile Rand
Buprenorphine 90 volumes of the following solution: dissolve 5.44 g of
potassium dihydrogen phosphate R in 900 mL of water R,
(Ph. Bur. motJOgraph 1180) adjust [Q pH 4.5 with a 5 per cent VIV solution of
phosphoric acid R and dilute to 1000 mL with water R;
- mobile phase B: acetonitrile R,
.oH
CH,
CH, (mlo) (per cent VIJI) (per cent V/J?
HO
CH, 0-2 8. II
2· 12 89 ..... 64 II ..... 36
12 - 15 64 --> 41 36 ..... 59
C"H.II NO, 467.6 52485-79-7
15 - 20 41 ..... 39 59 ..... 61
Action and use

Opioid receptor partial agonist; analgesic. Flow rate 1.3 mUmin.
Preparation Detection Spectrophotometer at 240 om.
Buprenorphine Transdennal Patches Injection 5 ~L.
PhE" ~ _ Identification of impurities Use the chromatogram supplied
with buprenorphine for system suitability CRS and the
DEFINITION chromatogram obtained with reference solution (b) to
(2S)-2-[17-(Cyclopropylmethyl)-4,5a-epoxy-3-hydroxy-6- identify the peaks due to impurities A, B, F, G, Hand J.
rnethoxy-e«, Jd-ethano-I 4a-morphinan-7a-ylj-3,3- Relative retention With reference to buprenorphine
dimethylbutan-2-ol. = =
(retention time about 8.5 min): impurity B about O.4j
Content impwity J = about I.I; impurity F = about 1.27;
98.5 per cent to 101.5 per cent (dried substance). impwity H = =
about 1.33; impwity A about 1.40;
impurity G = about 1.8.
CHARACTERS
Appearance System suitability Reference solution (b):
White or almost white, crystalline powder. - resolution: minimum 1.5 between the peaks due to
buprenorphine and impurity J.
Solub111ty
Very slightly soluble in water, freely soluble in acetone, Limits:
soluble in methanol, slightly soluble in cyclohexane. - correction factor: for the calculation of content, multiply the
It dissolves in dilute solutions of acids. peak area of impwity G by 0.3;
- impun'ty H: not more than 2.5 times the area of the
mp principal peak in the chromatogram obtained with
About 217 "C. reference solution (a) (0.25 per cent);
IDENTlFICATION - impurities A, BJ FJ J: for each impurity, not more than
Infrared absorption spectrophotometry (2.2.24). twice the area of the principal peak in the chromatogram
Comparison buprenorphine CRS. obtained with reference solution (a) (0.2 per cent);
__~.impun·ty G: not more than 1.5 times the area of the
Solution S reference solution (a) (0.15 per cent);
Dissolve 0.250 g in anhydrous uhand R and dilute to - unspecified impurities: for each impurity, not more than the
25.0 mL with the same solvent. area of the principal peak in the chromatogram obtained
Appearance of solution with reference solution (a) (0.10 per cent);
Solution S is clear (2.2.1) and colourless (2.2.2, Method II). - total: not more than 7 times the area of the principal peak
(0.7 per cent);
-103 (0 -107 (dried substance), determined on solution S.
Related substances the chromatogram obtained with reference solution (a)
Liquid chromatography (2.2.29). (0.05 per cent).
Test solution Dissolve 50.0 mg of the substance to be LOS8 on drying (2.2.32)
examined in methanolR and dilute [0 10.0 mL wilh the same Maximum 1.0 per cent, determined on 1.000 g by drying in
solvent. an oven at 105°C.
Reference solution (a) Dilute 1.0 mL of the test solution (Q
100.0 mL with methanolR. Dilute 1.0 mL of this solution to
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1-362 Buprenorphine 2022
ASSAY
Dissolve 0.400 g in 40 mL of anhydrous acetic acid R. Titrate
OH
with 0.1 M perchlon", acid, determining the end-point ; CH3
potentiometrically (2.2.20). CH,
H CH 3
I mL of 0.1 M perch/otic acid is equivalent to 46.76 mg of
OH
C29H.1NO•.
STORAGE E. (2S)-2-[17-(cyclopropylmethyl)-4,S«-epoxy-3,6-dihydroxy-
Protected from light. 6«, 14-ethano-14«-morphinan-7«-yl]- 3,3-dimethylbutan-2-
01 (6-Q-desmethylbuprenorphine),
IMPURITIES
Spedfiedimpurities A, B, F, G, H, J.
Other deuxtable impurities (!he following substances would, if N.~
CIi,
present at a sufficient level, be detecud by one or other of the tests CH,
in the monograph. They are limited by the general acceptance CH,
cntetion for otherhmspecified impun"ties and/or by thegeneral H CH3
monograph Substancesfor pharmaceutical use (2034). 1t is OCH3
therefore not n«tSsary to identify these impun"ties for
demonstration of compliance. See also5.10. Conrrol of impurities F. 17-(cyclopropylmethyl)-4,S«-epoxy-6-methoxy-7«-[I-(l,I-
in substances for phamuueutical use) C, D, E, I. dimethylethyljethenylj-Sa, 14-ethano-I4e<-morphinan-3-01,
A. (2S)-2-[17-(but-3-enyl)-4,S«-epoxy-3-hydroxy-6-methoxy-
G. 17,17/-di(cydopropylmethyl)-4,Sct;4',Set'-dlepoxy-?«, 7«'-
6o, 14-ethano-l4e<-morphinan-7«-yl]-3,3-dimethylbulan-2-
di [( IS) -I-hydroxy-I,2,2-trimethylpropyl]-6,6' -dimethoxy-
01,
2,2'-bi(oo,14-ethano-l4e<-morphinan)-3,3'-dlol (2,2'-
bibuprenorphine),
N~CH3
/H
H3C OH
"''\ ; CII,
.' CH3
. H CH)
OCH,
B. (2S)-2-(4,S«-epoxy-3-hydroxy-6-methoxy-6«, 14-ethano-
14e<-morphinan-7«-yl)-3,3-dimethylbutan- 2-01 H. (2S)-2-[17-butyl-4,S«-epoxy-3-hydroxy-6-methoxy-oo, 14-
(norbuprenorphine), ethano-I4e<-morphinan-7«-yl]3,3-dimethylbutan-2-01,
H3C OH
"'\ : CH,
: CH3
H CH3
H,CO OCH,
H,C
C. 4,S«-epoxy-7«-[(IS)-I-hydroxy-I,2,2-trimethylpropyl]-
3,6-dimethoxy-6a, 14-ethano-l4e<-morphinan-17- I. 17-(cyclopropylmethyl)-4",4"J5 fI ,5 11-tetramethyl-
carbonitrile, 4",S" dihydro-(7~H)-6a,14-ethano-(S~H)-
difurano[2 /,3',4',5':4,J2,J 3,5;2",3":6,7]-J4ct-morphinan-
3-01,
OH
" CH3
CH,
H CH3
OCH,
D. (2S)-2-[l7-(cyclopropylmethyl)-4,S«-epoxy-3,6-
dimethoxy-Sc,14-ethano-I4e<-morphinan-7«-yl]- 3,3- J. (2S)-2-[17-(cyclopropylmethyl)-4,S«-epoxy-3-bydroxy-6-
dimethylbutan-2-ol (3-Q-methylbuprenorphine), methoxy-Sc, 14-etheno-I4e<-morphinan-7«-yl]-3,3-
dimethylbutan-z-ol.
_____________ ~ PhE"
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2022 Buprenorphine Hydrochloride 1-363
Reference solution (b) Dissolve 5 mg of buprenorphine for

Buprenorphine Hydrochloride system suitability CRS (containing impurities AJ BJ F J GJ H
(Ph. Bur. monograph 118/) and D in 1.0 mL of methanol R.
Column:
- size: ,;:;; 0.05 m, (2) =4.6 mm;
- stationary phase: end__apped octad«y1silyl silica gelfor
chromatography R (3.5 urn);
• Hel
Mobile phase:
- mobile phase A: mix 10 volumes of acetonitrile Rand
90 volwnes of the following solution: dissolve 5.44 g of
504.1 53152-21-9 potassium dihydrogen phosphate R in 900 mL of waterR,
adjust to pH 4.5 with a 5 per cent VIV solution of
Action and use phosplwn·c acidR and dilute to 1000 mL with waterR;
Opioid receptor partial agonist; analgesic. - mobile phase B: acetonitrile R;
Preparations
Buprenorphine Injection Time Mobile phase A Mobile pbase B
(min) (per cent JlIJI) (per cent I'M
Buprenorphine Sublingual Tablets
0·2 80 II
PhE" _ 2 ~ 12 89 ---> 64 II -f 36
12 - 15 64 ---> 41 36 ---> 59
DEFINITION 15 - 20 41 -f 39 59 ---> 61
(2S)-2-[17-(Cyclopropylmethyl)-4,5"-epoxy-3-hydroxy-6-
methoxy-Se, Id-erhano-l ac-morphinan-ju-yf]-3,3-
dimethylbutan-z-ol hydrochloride.
Content
98.5 per cent to 101.5 per cent (dried substance). Injection 5 ~L
CHARACTERS
with buprenorphine for system suitability CRS and the
Appearance chromatogram obtained with reference solution (b) to
White or almost white, crystalline powder. identify the peaks due to impurities AJ BJ F J GJ H and J.
Solubility ReJauve retention With reference to buprenorphine
Sparingly soluble in water, freely soluble in methanol, soluble
in ethanol (96 per cent), practically insoluble in cyclohexane.
=
(retention time about 8.5 min): impurity B ;:;; about 0.4;
impurity J = about I.l; impurity F = about 1.27;
IDENTIFICATION =
impurity H about 1.33; impurity A abour 1.40; =
A. Infrared absorption spectrophotometry (2.2.24). impurity G ;:;; about 1.8.
Comparison buprenorphine hydrochloride CRS. System suitability Reference solution (b):
B. 3 mL of solution S (see Tests) gives reaction (a) of - resolution: minimum 1.5 between the peaks due to
chlorides (2.3.1). buprenorphine and impurity J.
Limits:
TESTS - correction factor. for the calculation of content) multiply the
Solution S peak area of impurity G by 0.3;
Dissolve 0.250 g in 5.0 mL of methanol R and, while stirring, - impurity H: not more than 2.5 times the area of the
dilute to 25.0 mL with carbon dioxide-free waterR. principal peak in the chromatogram obtained with
Appearance of solution reference solution (a) (0.25 per cent};
Solution S is clear (2.2.1) and colourless (2.2.2, Me/hod II). - impuriues A J B.J FJ J: for each impurity, not more than
Acidity or alkalinity twice the area of the principal peak in the chromatogram
To 10.0 mL of solution S add 0.05 mL of methyl red obtained with reference solution (a) (0.2 per cent);
solution R. Not more than 0.2 mL of 0.02 M sodium hydroxide - impwity· G: not more than 1.5 times the area of the
or 0.02 M hydrochloric acid is required to change the colour of principal peak in the chromatogram obtained with
the indicator. reference solution (3) (0.15 per cent);
- unspecified impun·ties: for each impurity, not more than the
area of the principal peak in me chromatogram obtained
Dissolve 0.200 g in methanol R and dilute to 20.0 mL with - total: not more than 7 times the area of the principal peak
the same solvent. in the chromatogram obtained with reference solution (a)
Related substances (0.7 per cent);
Liquid chromatography (2.2.29). - disregard limit: 0.5 times the area of the principal peak in
Test solution Dissolve 50.0 mg of the substance to be the chromatogram obtained with reference solutio_n (a)
examined in methanol R and dilute to 10.0 mL with the same (0.05 per cent).
. solvent. Loss on drying (2.2.32)
Reference solution (a) Dilute 1.0 mL of the test solution to Maximum 1.0 per cent) determined on 1.000 g by heating in
100.0 mL with methanolR. Dilute 1.0 mL of this solution to an oven at 115-120 "C,
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1-364 Buprenorphine Hydrochloride 2022
ASSAY
Dissolve 0.400 g in a mixture of S mL of 0.01 M hydroch/ori<
acid and SO mL of ethanol (96 per c<nu R. Carry out a
potentiometric titration (2.2. ZO), using 0.1 kl sodium
inflexion. Carry out a blanktitration.
1 mL of O. 1 M sodium hydroxide is equivalent to S0.41 mg of
E. (2S)- 2- [17-(cyclopropylmethyl)-4,S«-epoxy-3,6-dihydroxy-
C2,H,2CINO•.
6", l4-ethano-14«-morphinan-7«-yl]-3,3-dimethylbutan-2-
STORAGE 01 (6-G-desmethylbuprenorphine),
IMPURITIES
Specified impuniies A, B, F, G, H, J.
Otherdelectable impurities (the following ",bstances wau/d, if
cntetion for othethmspecified impurities and/or by the general
monograph Substances for pharmaceutical use (2034). It is F. 17-(cyclopropylmethyl)-4,S«-epoxy-6-methoxy-7«-[I-(I,1-
therefore not necessary w identify these impurities for dimethylethyI)e thenyf]- 6c<, 14-e thano-14«-morphinan-3-01,
demonstration of compliance. See also5.10. ContrOl of impurities
in substances for pharmaceutical use) C, D, E, 1.
G. 17)17'-di(cyclopropylmerhyl)-4,5a;4',5a' -diepoxy-?«,t«:
di [(I S)-l-hydroxy-I,2,2-trime thylpropyl]-6,6'-dimethoxy-
A. (2S)-2-[17-Ibut-3-enyI) -4, Sc-epoxy-3-hydroxy-6-methoxy-
2,2'-bi(6Cf:,14-ethano-l4ct-mOlphinan)-3,3'-diol (2,2'-
6", 14-ethano-14«-morphinan-7«-yl]-3,3-dimethylbutan-2-
bibuprenorphine),
01,
N~CH3
...H
H3C OH
---'\ .' CH,
... CH3
H cf<,
HO OCH,
H. (2S)-2-[17-butyl-4,S«-epoxy-3-hydroxy-6-methoxy-oo,14-
B. (2S)-2-( 4,S«-epoxy-a-hydroxy-ri-merhoxy-eu, l4-ethano- ethano-I4e<-morphinan-7«-yl]3,3-dimethylbutan-2-01,
14e<-morphinan-7«-yl)-3,3-dimethylbutan-2-01
(norbuprenorphine),
H3C OH CH,
.. ' \ .. CH,
.. CH3 CH,
H CH3 H,C
OCH,
T. 17-(cyclopropylmethyl)-4",4",5",5"-tetramethyl-
C. 4,S«-epoxy-7«-[(IS)-I-hydroxy-l ,2,2-trimethylpropyl]- 4",5" dihydr(}-(7~H)-oo,14-ethan(}-(S~H)-
3,6-dimethoxy-oo, l4-ethano-I4e<-morphinan-l7- difurano[2', 3',4',5/:4,12,13,5;2",3":6,7]-14ct.-morphinan-
carbonitrile, 3-01,
OH
: CH3
CH,
H CH 3
OCH3
J. (2S)-2-[ 17-(cyclopropylmethyl)-4,S«-epoxy-3-hydroxy-6-
D. (2S)-2-[ 17-(cyclopropylmethylj-d.Sa-epoxy-Sje- methoxy-Se, 14-etheno-I4e<-morphinan-7«-yl]-3,3-
dimethoxy-6«,14-ethano-I4e<-mo<phinan-7«-yl]-3,3- dimethylbutan-z-ol.
dimethylbutan-Z-cl (3-G-methylbuprenorphine), _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE",
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2022 Buserelin 1-365
Buserelin *** 0.9 to 1.1. Not more than traces of otheramino acids are
** ** present.
(Ph. Eur. monograph 1077) ***** TESTS
A 10 gIL solution is clear (2.2.1) and not more intensely
coloured man reference solution Y7 (2.2.2) Method11).
-58 to -49 (anhydrous, acetic acid-free substance),
determined on a 10 gIL solution.
1239 57982-77-1 Specific absorbance (2.2.25)
49 to 56, measured at the absorption maximum at 278 nm
Action and use (anhydrous, acetic acid-free substance).
Gonadotrophin releasing hormone (gonadorelin) analogue; Dissolve 10.0 mg in 100.0 mL of 0.01 M hydrochloric acid.
treatment of prostate cancer. Related substances
Pl>E<I _ Liquid chromatography (2.2.29).
DEFINITION
examined in 5.0 mL of the mobile phase.
5-0xo-L-prolyl-L-histidyl-L-tryptophyl-L-seryl-L-tyrosyl-O-
( I, I-dimethylethyl)-D-seryl-L-Ieucyl-L-arginyl-N-ethyl-L- Reference solution (aJ Dissolve the contents of a vial of
o-His-buserelin CRS (impurity A) in the mobile phase. Dilute
prolinamide.
an appropriate volume of this solution in the mobile phase to
Synthetic nonapeptide analogue of human gonadotrophin- obtain a final concentration of I mg/mL. Add 1.0 mL of the
releasing hormone GnRH with agonistic activity to test solution to 1.0 mL of this solution.
gonadorelin. It is obtained by chemical synthesis and is
available as an acetate.
busereJin CRS in the mobile phase. Dilute an
Content appropriate volume of this solution in the mobile phase to
95.0 per cent to 102.0 per cent (anhydrous, acetic acid-free obtain a final concentration of 1.0 mg/mL.
substance).
CHARACTERS 100.0 mL with the mobile phase.
Appearance Reference solution (d) Dissolve the contents of a vial of
White or slightly yellowish powder,hygroscopic. buserelin for peak idenrificarion CRS (containing
Solubility impurities F and G) in the mobile phase. Dilute an
Sparingly soluble in water and in dilute acids. appropriate volume of this solution in the mobile phase to
obtain a final concentration of I mglmL.
IDENTIFICATION
Carry out either tests A and B or tests A and C. Column:
- size: 1= 0.25 rn, 0 = 4 mm;
A. Examine me chromatograms obtained in the assay. - srationary phase: ocrade<y/silyl silica gelfor chromatography R
Results The principal peak in the chromatogram obtained (5 ~);
with the test solutionis similar in retention time and size to - temperature: 42 "C.
me principal peak in me chromatogram obtained with Mobile phase Mix 200 mL of acewnirrile Rand 700 mL of
reference solution (b). an 11.2 gIL solution of phosphoric acidR previously adjusted
B. Nuclear magnetic resonance spectrometry (2.2.64). to pH 2.5 with rriethylamine R.
Preparation 4 mg/mLsolution in a mixture of 20 volumes of Flow rate 0.8 mUmin.
deuterated acetic acid Rand 80 volumesof deuterium oxide R.
Comparison Dissolve the contents of a vial of buserelin for
Injection 10 J.lL of the test solution and reference
NMR identification CRS in a mixture of 20 volumes of solutions (a), (c) and (d).
deuterated acetic acidRand 80 volumes of deuterium oxide R to
obtain a concentration of 4 mglmL. Run time 60 min.
Operating conditions: Identification of impurities Use the chromatogram obtained
- field strength: minimum 300 MHz; with reference solution (a) to identify the peakdue to
- temperature: 27 °C. impurity A and the chromatogram obtained with reference
solution (d) to identify the peaksdue to impurities F and G.
Results Examine the 'H NMR specrrum from 0 to 9 ppm.
The lH NMR spectrum obtained is qualitatively similar to Relative retention With reference to buserelin (retention
the 'H NMR spectrum obtained with buserelinfor NMR = =
time about 23 min): impurity F about 0.83;
identification CRS. impurity A = about 0.91; impurity G = about 1.26.
C. Amino acid analysis (2.2.56). Method I for hydrolysis and System suitability Reference solution (a):
method 1 for analysis are suitable. - resolution: minimum J.5 between the peaks due to
impurity A and buserelin.
Express the content of each amino acid in moles. Calculate
the relative proportions of the amino acids, taking 1/6 of the Caku/ation of percentage contents:
sum of the number of moles of glutamic acid, histidine, - for each impurity, use the concentration of buserelin in
tyrosine, leucine, arginine and proline as equal to 1. reference solution (c).
The valuesfall within the following limits: serine 1.4 to 2.0j Limits:
proline 0.8 to 1.2; glutamic acid 0.9 to 1.1; leucine - impun·ty A: maximwn 2.5 per cent;
0.9 to 1.1; tyrosine 0.9 [0 l.lj histidine 0.9 to 1.Ij arginine - impurity F: maximum J.O per cent;
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1-366 Buspirone Hydrochloride 2022
He CH3
~ impurity G: maximum 1.0 per cent;
, --+-CH,
- unspecified impurities: for each impurity, maximwn 01 »<;
0.5 per cent; H-Trp-Ser-Tyr-D-Ser-leu- Arg-pro-~ CH3
- re[HJrlit/g threshold: 0.1 per cent. C. buserelin-(3-9)-peptide,
Acetic acid (2.5.34)
examined in a mixture of 5 volumes of mobile phase Band
95 volumes of mobile phase A and dilute 10 10.0 mL with
the same mixture of solvents.
Waler (2.5.12) D. [5-o-ryrosine]buserelin,
Maximwn 4.0 per cent) determined on 80.0 mg.
~
Bacterial endoroxlns (2.6.14)
o CH,
NH H3Ct CH,
of parenteral preparations without a further appropriate : 0 ~
procedure for the removal of bacterial endotoxins. H His-Trp-Ser-Tyr-D-Ser-leu-Arg-pro-~ CH3
ASSAY
o
Liquid chromatography (2.2.29) as described in the lest for E. [1-(5-oxo-o-proline)]buserelin,
Injection Test solution and reference solution (b). o
Calculate the percentage content of buserelin
(CroHS6NI6013) taking into account the assigned content of
CooHsoNI601J in buserelit/ CRS.
Y
H
. NH
o
H,C
01
CH,
'-?-C~
.n--His-Trp-ser-ser-Tyr-O-Ser'leu-Arg-pro-~ ~
CH 3
STORAGE
In an airtight container, protected from light, at a F. endo-3a-serine-buserelin,
temperature of 2 °C to 8°C. If the substance is sterile, the
LABELLING
The labelstares:
- the mass of peptide in the container;
- where applicable, that the substance is suitable for use in
the manufacture of parenteral preparations.
G. endo-8a-proline-buserelin.
IMPURlTffiS _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw
Specified impurities A, F, G.
Otherdetectable impurities (thefollowiug substat/ces would, if
it/ the monograph. They arelimiud by the general acceptance Buspirone Hydrochloride
criterion for otherlunspecified impurities ondlor by the general
monograph Substat/ces for pharmaceutical use (2034). It is (Ph. Eur. monograph 1711)
demonstratum of compliance. See also 5.10. Control of impuniies
in substancesfor pharmaceutical use) B, C, D, E.
• HCI
Y
H
\
o
O
NH H3Ct CH,
0
CH,
!rD-HIS-Trp-Ser-Tyr-lH)ef-LeU-Arg-Pro-~
»<;
CH3
422.0 33386-08-2
A. [2-o-rustidine)buserelin, Action and use
Non-benzodiazepine hypnotic; treatment of anxiety.
o
Y
PhEIJf _
H H,CtCH,
CH,
DEFINITION
... 0 ...............
H n-HIS-TrP-O-Ser-Tyr-o-Ser-Leu-Arg-Pro-~ CH3 8- [4- [4-(Pyrimidin-2-yl)piperazin-l-yl] butyll-a-ezaepiro
o [4.5]decane-7,9-<lione hydrochloride.
Content
B. [4-n-serine]busere1inl 99.0 per cent CO 101.0 per cent (dried substance).
CHARACTERS
Appearance
White or ahnost white, crystalline powder.
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2022 Buspirone Hydrochloride 1-367
Solubility impurity D = about 0.7; impurity E = about 0.8;

Freely soluble in water and in methanol, practically insoluble impurity F : : : about 0.9; impurity G ;;;; about 1.05;
in acetone. = =
impurity H about 1.1; impurity I about 1.2;
It shows polymorphism (5.9). impurity J = about 1.5.
IDENTIFICATION Relative retention at 210 nm With reference to buspirone
(retention time = about 25 min): impurity K = about 0.6;
A. Inflated absorption spectrophotometry (2.2.24).
impurity L = about 1.7; impurity M = about 1.8;
Comparison buspirone hydrochloride CRS. impurity N = about I.9.
If the spectra obtained in the solid state show differences, System suitability Reference solution (b):
dissolve the substance to be examined and the reference - peak-w-vaJley ratio at 240 nm: minimum 5.0, where
substance separately in methanol R, evaporate to dryness on a
water-bath and record new spectra using the residues.
=
Hp height above the baseline of the peak due to
impurity G and H; = height above the baseline of the
B. It gives reaction (a) of chlorides (2.3.1). lowest point of me curve separating this peak from the
TESTS peak due to buspirone;
Related substances - resolution at 210 nm: minimum 4.0 between the peaks due
Liquid chromatography (2.2.29). to impurity L and impurity N;
- the chromatograms obtained are similar to the
Testsolution Dissolve 25.0 mg of the substance to be chromatograms supplied with buspirone for system
suitability CRS.
mobile phase A.
Limits Spectrophotometer at 240 nm:
Reference solution (aJ Dilute 1.0 mL of the test solution to - correction factor. for the calculation of content, multiply the
peak area of impurity J by 2,
- impurity E: not more than 3 times the area of the principal
Reference solution (b) Dissolve the contents of a vial of peak in me chromatogram obtained with reference
bwpirone for system suitability CRS (containing impurities E, solution (a) (0.3 per cent),
G, J, Land N) in 2.0 mL of mobile phase A and sonicate for - impun°ty J: not more than twice the area of the principal
10 min. peak in me chromatogram obtained with reference
Column: solution (a) (0.2 per cent),
- size: 1= 0.15 m, 0;;;; 406rnm, - any other impurity: for each impurity, not more than the
- stationary phase: tXtade<y1sl1yl silica gelfor chromatography R area of the principal peak in the chromatogram obtained
(5 urn), with reference solution (a) (0.1 per cent),
- temperature: 40 °C. - total: not more than 4 times the area of the principal peak
Mobile phase: in the chromatogram obtained with reference solution (a)
- mobile phase A: mix 950 volumes of a solution containing (0.4 per cent),
6.8 gIL of potassium dihydrogen phosphate R and 0.93 gIL ---: disregard limit: 0.5 times the area of the principal peak in
of sodium hexanesulfonate monohydrate R, previously the chromatogram obtained with reference
adjusted to pH 3.4 with plwsphoric acid Rand 50 volumes solution (a) (0.05 per cent).
of aUlOnitn·le R1; Limits Spectrophotometer at 210 nm:
- mobile phase B: mix 250 volumes of a solution comaining - impuniy K: not more than the area of the principal peak
3.4 gIL of potassium dihydrogen phosphate R and 3.52 gIL in the chromatogram obtained with reference solution (a)
of sodium hexanesulfonate monohydrate R, previously (0.1 per cent),
adjusted to pH 2.2 with phosphon·c acid Rand - any otherimpun·'y eluting Wilh a relative retention greater
750 volwnes of acetonitrile R1; than 1.6: for each impurity, not more than the area of the
Time Mobile phase A Mobile phase B reference solution (a) (0.1 per cent),
(min) (per cent VJ1') (per cent VIJi) - total: not more than twice the area of the principal peak in
0-. 90 10 the chromatogram obtained with reference solution (a)
6 - 34 90 --> 42 10 .... 58 (0.2 per cent),
34 - 45 42 58 - disregard limit: 0.5 times the area of the principal peak in
45 - 55 42 -+ 0 58 .... 100 the chromatogram obtained with reference solution (a)
55 - 56 0--> 100 100 -+ 0 (0.05 per cent).
56 - 60 100 0 Loss on drying (2.2.32)
60 - 61 Joo .... 90 0-+ 10
Maximum 0.5 per cent, determined on 1.000 g ~ drying at
105 "C.
Flow rate I mUmin. Sulfated ash (2.4.14)
Detection Variable wavelength spectrophotometer capable of Maximum 0.1 per cent, determined on 1.0 g.
operating at 240 nm and at 210 nm.
ASSAY
Injection 20 ~L. Dissolve 0.150 g in 10 mL of glacial acetic acidR and add
Identification of impurities Use the chromatogram supplied 50 mL of acetic anhydride R. Titrate with 0.1 M perchloric
with buspirone for system 'uitability CRS and the acid, determining the end-point potentiometrically (2.2.20).
chromatogram obtained with reference solution (b) to 1 mL of 0.1 1\1 perchloric acid is equivalent to 21.10 mg
identify the peaks due to impurities E, G, J, L and N.
of C21H32CINj02'
Relative retention at 240 nm With reference to buspirone
=
(retention time = about 25 min): impurity A about 0.2; STORAGE
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1-368 Buspirone Hydrochloride 2022
IMPURITIES
Spedfied impurities E, J, K.
Otherdetectable impurities (the following substances would, .f
present at a sufficient level, be duetted by oneor other of the tests
in the monograph. They an! limited by the general acaptonce
cntetion for other/unspecified impun"ties and/or by the general
monograph Substanasfor pharmaceutical use (2034). It is G. 2,2 /_(piperazine-l,4-diyl)dipyrimidine,
therefore not neussary to identify these impun"u'es for
in substances/or phannauutiall use) A, B.I C, D, F, GJ H, I,
L,M, N.
A. 2-(pipernzin-I-yl)pyrimidine,
H. bis[4-[4-(pyrimidin-2-yl)piperazin-l-yl]butyl)
(cyclopenrane-I, I-diyl)diacetate,
B. 8_(pyrimidin_2_yl)_8_aza_5_azoniaspiro[4.5)decane,
J. 8-[4-[4-(5-chloropyrintidin-2-yl)piperazin-I-yl)butyl]-8-
azaspiro[4.5]decane-7,9-dione,
~
H
N~Nl
° 0 0 ~N"JN~
o~~
C. 2,2'-[butane-I,4-diylbis(pipernzine-1,4-<1iyl)]dipyrimidine,
N-",
rN~O~Nl
C
I
NyN ~
.."N
~N"JN~
N -'" J. 4-(7,9-dioxo-8-azaspiro[4.5]dec-8-yl)butyl [l-[2-oxo-2-
[[4-[4-(Pyrimidin-2-yl)pipernzin-I-yl] butyl)amino]
D.2,2'_[oxybis[butane_I,4_diyl(pipernzine_I,4-<1iyl»)] ethyl]cyclopentyl] acetare,
CQ:0
dipyrimidine,
K. 8_azaspiro[4.5]decane-7,9-dione,
C1l~CI
E. [1-[2-oxo-2-[[4-[4-(Pyrimidin-2-yl)piperazin-I-yl)butyl]
amino)ethyl)cyclopentyl]acetic acid,
L. 8-(4-cblorobutyl)-8-azaspiro[4.5)decane-7,9-<1ione,
F. 4_[4_(Pyrimidin_2_yl)pipernzin_l_yl)butyl [1-[2-oxo-2-[[4-
C1l~B' o
[4-(pyrimidin-2-yl)piperazin-I-yI]butyl]amino)ethyl]
M.8_(4-bromobutyl)-8-azaspiro[4.5)decane-7,9-dione,
cyclopentyl]acetate,
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2022 Butyl Hydroxybenzoate 1-369
C. To 0.1 g add 5 mL of I M sodium hydroxide. Heat until a

clear solution is obtained. Allow to cool. To 2 mL of the
solution add 0.1 mL of potassium pennanganau solution R.
The colour changes from purple through violet to blue and
finally to green. Filter and add I mL of ammoniacal silver
nitrate solution R. A precipitate is formed.
N. 8,8'-(butane-l ,4-diyl)bis(8-azaspiro[4.5)decane-7,9- D. To 0.1 g add 0.1 g of potassium nitrate Rand 0.25 g of
dione). sodium hydroxide R, mix and heat to fusion. Allow to cool and
_ _ _ _ _ _ _ _ _ _ _ _. F'llE1r
dissolve the residue in 5 mL of water R. Adjust to pH 1-2
using dilute hydrochloric acid R. The solution gives reaction (a)
of sulfates (2.3. I).
TESTS
Busulfan Appearance of solution
(Ph. Eur. monograph 0542) than reference solution B 7 (2.2.2) 1'Ilerhod II).
o.. 0
H3C/S ...O~O ...s/CH3

Dissolve 0.25 g in 20 rnL of acetonitrile R, dilute to 25 mL
with water R and examine immediately.
Acidity
o'"0 Dissolve 0.20 g with heating in 50 mL of anhydrous
ethanol R. Add 0.1 mL of methylredsolution R. Not more
246.3 55-98-1 than 0.05 mL of O. I M sodium hydroxide is required to
change the colour of the ~dicator.
Action and use Loss on drying (2.2.32)
Cytotoxic alkylating agent. Maximum 2.0 per cent, determined on 1.000 g by drying in
Preparation at 60°C.
tlCUUQ
Busulfan Tablets Sulfated ash (2.4.14)
I'OE,; _ Maximum 0.1 per cent, determinedon 1.0 g.
DEFINITION ASSAY
Butane-l,4-diyl di(methanesuJfonate). To 0.250 g add 50 mL of water R. Shake. Boil under a reflux
condenser for 30 min and, if necessary, make up to the
Content initial volume with water R. Allow to cool. Using 0.3 mL of
99.0 per cent to 100.5 per cent (dried substance). phenolphthalein solution R as indicator, titrate with 0.1 M
CHARACTERS sodium hydroxide until a pink colour is obtained.
Appearance I mL of 0.1 M sodium hydroxide is equivalent to 12.32 mg of
White or almost white, crystalline powder. CJI I4O.S,.
Solublllty STORAGE
Very slightly soluble in water, freely soluble in acetone and in In an airtight container, protected from light.
acetonitrile, very sllghtly soluble in ethanol (96 per cent). _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'OEII
mp
About 116 'C.
IDENTIFICATION
First identification: A. Butyl Hydroxybenzoate
Burylparaben
(Butyl Parahydroxybenzoate, Ph. Eur. monograph 0881)
Comparison busulfan CRS.
B. Thin-layer chromatography (2.2.27). o
examined in 2 mL of acetone R.
~O~CH3
Reference solution Dissolve 20 mg of busulfan CRS in 2 mL Ho N
of cuerone R.
Plate TLC silica gel G plateR. 194.2 94-26-8
Mobile phase acetcne R, toluene R (50:50 VIV). Action and use
Application 5 ~L. Excipient.
Development Over a path of 15 em. I'OE,; ~ --
Drying In a currentof wann air.
Deteaion Spray with anisaldehyde solution R and heat at DEFINlTION
120 'C. Butyl 4-hydroxybenzoate.
Results The principal spot in the chromatogram obtained Content
with the test solution is similar in position, colour and size to 98.0 per cent to 102.0 per cent.
reference solution.
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1-370 Butyl Hydroxybenzoate 2022
CHARACTERS Reference solution (a) Dissolve 5 mg of 4-hydroxybenzm'c

Appearance acid R (impurity A), 5 mg of propylparahydroxybenzoau R
White or almost white, crystalline powder or colourless (impurity D) and 5 mg of the substance to be examined in
crystals. the mobile phaseand dilute to 100.0 mL with the mobile
Solubility phase. Dilute 1.0 mL of the solution to 10.0 mL with the
Very slightly soluble in water, freely solublein ethanol mobile phase.
(96 per cent) and in methanol. Reference solution (b) Dissolve 50.0 mg of butyl
parahydroxybenzoal< CRS in 2.5 mL of methanol R and dilute
IDENTIFICATION
1050.0 mL with the mobile phase. Dilute 10.0 mL of the
First identification: A, B. solution to 100.0 mL with the mobile phase.
Second identification: A, C. Reference solution (c) Dilute 1.0 mL of the lest solution 10
A. Melting point (2.2.14): 68 °C to 71 °C. 20.0 mL with the mobile phase. Dilute 1.0 mL of this
B. Infrared absorption spectrophotometry (2.2.24). solution to 10.0 mL with the mobilephase.
Comparison butylparahydroxybeuzoal< CRS. Reference solution (d) Dissolve 5 mg of butyl
C. Thin-layer chromatography (2.2.27). parahydroxybenzoal< impurityE CRS (iso-butyl
parahydroxybenzoate) in the mobilephase and dilute to
Testsolution (a) Dissolve 0.10 g of the substance [Q be
examined in acewne R and dilute to 10 mL with the same
solvent. Reference solution (e) Dilute 0.5 mL of reference solution (d)
to 50.0 mL with reference solution (b).
Te.stsolution (b) Dilute I mL of test solution (a) to 10 mL
with acetom R. Column:
Reference solution (a) Dissolve 10 mg of butyl
- size: 1= 0.15 m, 0 == 4.6 mm;
- sratwnary phase: end-eapped oczadecylsiJyl silica gelfor
parahydroxybenzoate CRS in acetone R and dilute to 10 mL
Reference solution (b) Dissolve 10 mg of propyl
MobUe phase 6.8 gIL solution of potassium dihydrogen
parahydroxybenzooee R in 1 mL of test solution (a) and dilute
phosphate R. methanol R (50:50 VIV).
to 10 mL with acetone R.
Pial< TLC oetade<ylsilyl suica gel F254 pial< R.
Mobile phase glacialacetic acid R, waterR, methanol R
(1:30:70 VIVW). Injection 10 JlL of the test solutionand reference
solutions (a), (c) and (e).
Applicatian 2 J.1L of test solution (b) and reference
solutions (a) and (b). Run time 1.5 times the retention time of butyl
parahydroxybenzoate.
Identification 0/ impurities Use the chromatogram obtained
Drying In air.
with reference solution (a) to identify the peaks due to
Detection Examine in ultraviolet light at 254 nm. impurities A and D; use the chromatogram obtained with.
System mizabilily Reference solution (b): reference solution (e) to identify the peakdue to impurity E.
- the chromatogram shows 2 clearly separated principal Relativeretention Withreference to butyl
spots. parahydroxybenzoate (retention time = about 22 min):
Results The principal spot in the chromatogram obtained impurity A == about 0.1; impurity D = about 0.5;
with test solution (b) is similar in position and size to the impurity E = about 0.9.
principal spot in the chromatogram obtained with reference System mirability:
solution (a). - resolution:
TESTS - minimum 5.0 between the peaks due to impurity D
Solution S and butyl parahydroxybenzoate in the chromatogram
Dissolve 1.0 g in ethanol (96 per cent) R and dilute 10 obtained withreference solution (a);
10 mL with the same solvent. - minimum 1.5 between the peaks due to impurity E
Appearance of solution and butyl parahydroxybenzoate in the chromatogram
Solution S is clear (2.2.1) and not moreintensely coloured obtained with reference solution (e).
thanreference solutionBY6 (2.2.2, JHethod II). Limits:
- correction factor. for the calculation of content, multiply the
Acidity
peak area of impurity A by 1.4;
To 2 mL of solution S add 3 mL of ethanol (96 per cent) R,
- impudty A: not more than the area of the principal peak
5 mL of carbon dioxide-free water R and 0.1 mL of bromocresol
in the chromatogram obtained withreference solution(c)
green solurion R. Not more than 0.1 mL of 0.1 M sodium
(0.5 per cent);
hydroxide is required to change the colour of the indicator to
blue.
Related substances with reference solution (c) (0.5 per cent);
Liquid chromatography (2.2.29). - totol: not more than twice the area of the principal peak in
Test sdiuion Dissolve 50.0 mg of the substance to be the chromatogram obtained withreference solution (c)
examined in 2.5 mL of methanol R and dilute to 50.0 mL (1.0 per cent);
with the mobile phase. Dilute 10.0 mL of the solution to - disregard limit: 0.2 times the area of the principal peak in
100.0 mL with the mobile phase. the chromatogram obtained with reference solution (c)
(0.1 per cent).
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2022 Butylated Hydroxyanisole 1-371

Butylated Hydroxyanisole ***
Maximum 0.1 per cent, determined on 1.0 g. *** ***
ASSAY (Bury/hydroxanisole, Ph. Bur. monograph 0880) ***
Injection Test solution and reference solution (b).
Calculate the percentage content of C llH140J from the
declared content of butyl parahydroxybenzoale CRS.
IMPURITIES
180.3 25013-16-5
Oiherdetectable impurities (thefollowing subS/ances would, if Action and use
present at a sufficient/wel, be detected by one or other of the tests Antioxidant.
in the monograph. They arelimited by thegeneral acapumce PhE" _
criterion for orher/unspecified impuniies and/or I>y ehe general
monograph Substances for pharmaceutical use (2034). Ie is DEFINITION
there/ore not neassmy to identify these impwilieJ for Butylhydroxyanisole is 2-( I, l-dimethylethyl)-4-
demonstration of compliance. See also5.10. Control 0/ impurities methoxyphenol containing not more than 10 per cent of
in substances for pharmaceutical use) B, C, D, E. 3-(I,I-dirnethylethyl)-4-methoxyphenol.
f"/TCO,H CHARACTERS
A white, yellowish or slighdy pinkish, crystalline powder,
HO~ practically insoluble in water, verysoluble jn methylene
chloride, freely soluble in alcohol and in fatty oils, It dissolves
in dilute solutions of alkali hydroxides,
A. 4-hydroxybenzoic acid,
IDENTIFICATION
o A. Examine the chromatograms obtained in me test for
HO
d""
I 0
' CH' related substances. The principal spot in the chromatogram
and size to the principal spot in the chromatogram obtained
B. methyl 4-hydroxybenzoate (methyl parabydroxybenzoate), B. To 0.5 mL of solution S (see Tests) add 10 mL of
aminopyrazolone solution R and 1 mL of potassium /erricyanide
solution R. Mix and add 10 mL of methylene chloride R. Shake
vigorously. Afterseparation, the organic layer is red.
C. Dissolve about 10 mg in 2 mL of alcohol R. Add I mL of
a I gIL solutionof testosterone propionau R In akohol Rand
2 mL of di/uu sodium hydroxide solution R. Heat in a water-
C. ethyl 4-hy<iroxybenzoace (ethyl parahydroxybenzoare), bath at 80 °C for 10 min and aUow to cool. A red colour
develops.
o TESTS
d
~ CH' Solution S
I 0
Dissolve 2.5 g in alcohol R and dilute to 25 mL with the
HO "" same solvent.
D. propyl 4-hydroxybenzoate (propyl parahydrcxybenzoate),
o than intensity 5 of the range of reference solutions of the
most appropriate colour (2.2.2.1 Method 11).
HO d~..."". I
° - " ' yCH,
CH
3
Related substances
gel GRas the coatingsubstance.
E. 2-methylpropyl -l-hydroxybenzoate (iso-butyl Tese solution (a) Dissolve 0.25 g of the substance to be
parahydroxybenzoate). examined in methylene chloride R and dilute to 10 mL with
_ _ _ _ _ _ _ _ _ _ _~ PhE"
the same solvent.
Test sduuon (1)) Dilute I mLoftestsolution (a) to 10 mL
with methylene chloride R.
Reference solution (a) Dissolve 25 mg of
bury/hydroxyanisole CRS in meehylene chloride R and dilute to
Reference solution (b) Dilute I mL of reference solution (a)
to 20 mL with methylene chloride R.
Reference sdution (c) Dissolve 50 mg of hydroquinone R in
5 mL of alcohol R and dilute to 100 mL with methylene
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1-372 Butylated Hydroxytoluene 2022
chloride R. Dilute 1 mL of this solution to 10 mL with (2.2.25)) the solution shows an absorption maximum at
methylene chloride R. 278 nm. The specific absorbance at the maximum is 80 to
Apply separately to the plate 5 ~ of each solution. Develop 90.
over a path of 10 em using methylene chloride R. Allow the C. Examine by infrared absorption spectrophotometry
plate to dry in air and spray with a freshly prepared mixture (2.2.24») comparing with the spectrum obtained with
of 10 volumes of potassium fenicyanide solutUm R, 20 volumes butylhydroxytoluene CRS.
of/erricchloride solution Rl and 70 volumes of waterR. In the D. Dissolve about 10 mg in 2 mL of alcohol R. Add I mL of
chromatogram obtained with test solution (a): any violet-blue a I gIL solutionof testasterone propionate R in akohol R and
spot with an RF value of about 0.35 (corresponding to 2 mL of dilute sodium hydroxide solution R. Heat in a water-
3-(I,I-dimethylethyl)-4-methoxyphenol) is not mote intense bath at 80°C for 10 min and allow to cool. A blue colour
man the principal spot in the chromatogram obtained with develops.
reference solution (a) (10 per cent); any spot corresponding
TESTS
to hydroquinone is not more intense than the principal spot
in the chromatogram obtained with reference solution (c) Appearance of soludon
(0.2 per cent); any spot, apart from the principal spot and Dissolve 1.0 g in methanol R and dilute to 10 mL with the
any spots corresponding to 3-(I,I-dimethylethyl}-4- same solvent. The solutionis clear (2.2.1) and not more
methoxyphenol and hydroquinone, is not more intense than intensely coloured than reference solution Y j or BYj (2.2.2,
the principal spot in the chromatogram obtainedwith MethodII).
reference solution (b) (0.5 per cent). Freezing-point (2.2.18)
Sulfated ash (2.4.14) 69 °C to 70 °C.
Not more than 0.1 per cent, determined on 1.0 g. Related substances
STORAGE
gd GRas the coatingsubstance.
Store protected from light.
Test solution Dissolve 0.2 g of the substance to be examined
IMPURITIES in methanol R and dilute to 10.0 mL with the same solvent.
Reference sduiion Dilute 1 mL of the test solution to
~OH 200 mL with methanol R.
HO~ Apply separately to the plate 10 J.lL of each solution. Develop
over a path of IS em using methylene chloride R. Dry the plate
A. benzene-I,4-diol (hydroqujnone). in air and spray with a fresWy prepared mixture of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE,;
10 volumes of potassium fenicyanide solution R, 20 volumes of
feme chloride solutUm Rl and 70 volumes of waterR. Any spot
in the chromatogram obtained with the test solution) apart
from the principal spot, is not more intense than the spot in
Butylated Hydroxytoluene the chromatogram obtainedwith the reference solution
(0.5 pet cent).
(Butylhydroxytoluene, Ph. Eur. monograph 0581)
Not more than 0.1 per cent) determined on 1.0 g.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Cabergoline
220.4 /28-37-0
Action and use

Antioxidant
PhE" _
DEFINITION
Butylhydroxytoluene is 2,6-bis(I,I-dimethylethyl)-4-
methylphenol.
CHARACTERS 45\.6 81409-9().J
A white or yellowish-white, crystalline powder, practically
insoluble in water, verysolublein acetone, freely soluble in Action and use
alcohol and in vegetable oils. Dopamine D2 receptor agonist.
Preparation
IDENfIFlCATION
Cabergoline Tablets
Second identification: A, B, D. PhE" _
A. Freezing-point (see Tests). DEFINITION

B. Dissolve 0.500 g in ethanol R and dilute to 100.0 mL with 1-Ethyl- 3- [3-(dimethylamino)propyl] -3-[ [(6aR,9R, IOaR)-7-
the same solvent. Dilute 1.0 mL of this solution to 100.0 mL (ptop-2-enyl) -4, 6,6a, 7 ,8,9, I 0, I Oa-octahydroindolo[4,3-fg]
with ethanol R. Examined between 230 ron and 300 nm quinolin-9-yl]carbonyl] urea.
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2022 Cabergoline 1-373
Content System suitability Reference solution (c):

98.0 per cent to 102.0 per cent (anhydrous substance). - resolution: minimum 3,0 between the peaks due to
CHARACTERS cabergoline and impurity A.
Appearance Limits:
White or almost white, crystalline powder. - impurities A, C: for each impurity, not more than
1.5 rimes the area of the principal peak in the
Solubility
PracticaUy insoluble in water, freely soluble in ethanol (0.3 per cent);
(96 per cent), very slightly soluble in hexane. It is slightly
- impurities BJ D: for each impurity, not more than
soluble in 0.1 M hydrochloric acid.
It shows polymorphism (5.9). chromatogram obtainedwith reference solution (b)
IDENTIFICATION (0.1 per cent);
A. Specific optical rotation (see Tests). - any other impun'ty: for each impurity, not more than
B. Infrared absorption spectrophotometry (2.2.24). 0.5 times the area of the principal peak in the
Comparison cabergoline CRS. (0.1 per cent);
If the spectra obtained in the solid state show differences, - wtal: not more than 4 times the area of the principal peak
dissolve 50 mg of the substance to be examined and 50 mg in the chromatogram obtained with reference solution (b)
of the reference substance separately in I rnL of ethanol (0.8 per cent);
(96 percent) R, evaporate (0 dryness and record new spectra - disregard limit: 0.25 times the area of the principal peak in
using the residues. the chromatogram obtained with reference solution (b)
TESTS (0.05 per cent).
Specific optical rotation (2.2.7) Water (2.5.1Z)
-77 to -83 (anhydrous substance). Maximum 0.5 per cent, determined on 1.000 g.
Dissolve 0.100 g in ethanol (96 percent) Rand dilute to ASSAY
50.0 mL with the same solvent. Liquid chromatography (2.2.29) as described in the test for
Related substances related substances with the following modification.
liquid chromatography (2.2.29). Prepare the solutions Injection Test solution and reference solution (a).
immediately before use and protected from light. Calculate the percentage content of C2JI37NS02 from the
Testsolution Dissolve 30.0 mg of the substance to be areas of the peaks and the declared content of
examined in the mobilephase and dilute to 25.0 mL with cabergo/ine CRS.
the mobile phase.
STORAGE
Reference solution (a) Dissolve 30.0 mg of cabergoline CRS in
the mobile phaseand dilute to 25.0 mL with the mobile
phase. IMPURITIES
Reference solution (b) Dilute 1.0 mL of the test solution to Spedfied impurities A, B, CJ D.
Reference solution (c) Suspend 50 mg of the substance to be
examined in 10 mL of 0.1 M sodium hydroxide. Stir for about
15 min. To I mL of the suspension add I mL of 0.1 M
hydrochloric acid and dilute to 10 mL with the mobile phase.
Sonicate until dissolution is complete. The main degradation
productobtained is impurity A.
Column:
A. (6aR,9R,1 OaR)-7-(prop-2-enyl)-4,6,6a,7,8,9,10, 10a-
- size: 1= 0.25 m, 0 = 4.6 mm,
oetahydroindolo[4,3-fg]quinoline-9-carboxylic acid,
- stationary phase: octade,;ylsi!YI silica gelfor chromalOgraphy R
(10 urn).
J.\tIobile phase Mix 16 volumesof acetomuile Rand
84 volumes of a freshly prepared 6.8 gIL solution of
pH 2.0 with plwsphoric acid R. Add 0.2 volumes of
methylamine R.
Injection 20 JlL of the test solution and reference B. (6aR,9R, 10aR)-N"-[3-(dimethylamino)propylj-N'-ethyl-7-
solutions (b) and (c). (prop-2-enyl)-6a,7,8,9,10,1 Oa-hexahydroindolo[4,3-fg]
Run time 4 times the retention time of cabergoline. quinoline-4,9(6H)-dicarboxamide,
Relative retention With reference to cabergoline (retention
time = about 12 min): impurity D =about 0.3;
impurity B = about 0.6; impurity A = about 0.8;
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1-374 Caffeine 2022
IDENTIFICATION
First identification: A, B. E.
Comparison caffeine CRS.
C. To 2 mL oCa saturated solutionadd 0.05 mL of iodinated
C. (6aR,9R, IOaR)-N"-[3-(dimethylamino)propyl]-N'-ethyl- potassium iodide solution R. The solutionremains clear.
N"-(ethylcarbamoyl)-1-(prop-2-enyl)-6a,1,8,9,10,10a- Add 0.1 mL of dilute hydrochloric acid R; a brown precipitate
heKahydroindolo[4,3-fgjquinoline-4,9(6H)-dicarboKamide, is formed. Neutralise with dilute sodium hydroxide solution Rj
the precipitate dissolves.
D. In a ground-glass-stoppered tube, dissolve about 10 mg in
0.25 mL of a mixture of 0.5 mL of acetylacetone Rand 5 mL
of dilute sodium hydroxide solution R. Heat in a water-bath at
80 'C for 1 min. Cool and add 0.5 mL of
dimethylaminobenzaldehyde solution R2. Heat again in a water-
bath at 80 'C for 1 min. Allow to cool and add 10 mL of
water R; an intense blue colour develops.
D. (6aR,9R,IOaR)-N-[3-(dimethylamino)propyl]-1-(prop-2- E. Loss on drying (see Tests).
enyl)-4,6,6a,1,8,9, I0, I Oa-oetahydroindolo [4,3-fg] F. It gives the reaction of xanthines (2.3.1).
quinoline-9-carboxamide.
TESTS
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pl>Eor
Solution S
water R prepared from distifhd water R, cool and dilute to
50 mL with me same solvent.
Caffeine Appearance of solution
Solution S is clear (2.2.1) and colourless (2.2.2, Method If).
Anhydrous Caffeine
Acidity
solution Rl; the solution is green or yellow. Not more than
0.2 mL of 0.01 M sodium hydroxide is required to change the
colour of the indicator to blue.
Related substances
Test solution Dissolve 0.100 g oCthesubstance to be
194.2 58-08-2 examinedin the mobile phase and dilute to 50.0 mL with
the mobile phase. Dilute 1.0 mL of this solution to 10.0 mL
Action and use with the mobile phase.
Central neLVOUS system stimulant. Reference solution (a) Dilute 2.0 mL of the test solution to
Preparations 100.0 mL with the mobile phase. Dilute 1.0 mL of this
Aspirin and Caffeine Tablets solution to 10.0 mL with the mobile phase.
Caffeine Citrate Injection Reference solution (b) Dissolve 5 mg of caffeine for system
Caffeine Citrate Oral Solution suitability CRS (containing impurities A, C, D and F) in the
Paracetamol and Caffeine Capsules mobile phase and dilute to 5 mL with the mobile phase.
Dilute 2 mL of this solution to 10 mL with the mobile
Paracetamol and Caffeine Tablets
phase.
Paracetamol, Codeine Phosphate and Caffeine Capsules
Column:
Paracetamcl, Codeine Phosphate and Caffeine Tablets - size: (= 0.15 m, 0 = 4.6 mm;
Pl>Eor _ - stationary phase: end-capped amidohexaclecylsi/yl silica gelfor
chromaI<Jgraphy R (5 urn).
DEFINITlON Mobile phase Mix 20 volumes of lelrahydrofuron R,
1,3,1-Trimethyl-3,1-dibydro-IH-purine-2,6-dione. 25 volumes of acetonitrile Rand 955 volumesof a solution
Content containing 0.82 gIL of anhydrous sodium acetate R previously
98.5 per cent to 101.5 per cent (dried substance). adjusted to pH 4.5 with glacial acetic acid R.
CHARACTERS Flow rate 1.0 mUmin.
Appearance Detection Spectrophotometer at 275 run.
White or almost white, crystalline powder or silkycrystals. Injection I 0 ~L.
Solubility Run time 1.5 times the retention time of caffeine.
Sparingly soluble in water, freely soluble in boiling water, Identification of impun'lies Use the chromatogram supplied
slightly soluble in ethanol (96 per cent). It dissolves in with caffeine for system suitability CRS and the chromatogram
concentrated solutions of alkali benzcates or salicylares.
It sublimes readily.
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2022 Caffeine Hydrate 1-375
obtained with reference solution (b) to identify the peaks due

to impurities A, C, D and F.
Relativeretention With reference to caffeine (retention
impurity D = about 0.42; impurity F = about 0.6j
=
System suitability Reference solution (b): C. 1,3,9-trimethyl-3,9-dihydro-IH-putine-2,6-dione
(isocalfeine),
- resolution: minimum 2.0 between the peaksdue (0
impurities C and D and minimum 2.5 between the peaks
due £0 impurities F and A.
Limits:
- unspecified impurities; for each impurity, not more than
0.5 times me area of the principal peak in the
(0.10 per cent);
D. 3,7-dimethyl-3,7-<1ihydro-IH-putine-2,6-<1ione
- total: not more than 0.5 times the area of the principal (theobromine),
the chromatogram obtained wirh reference solution (a)
(0.05 per cent).
Sulfates (2.4.13)
Maximum 500 ppm, determined on 15 mL of solution S.
Prepare the standard using a mixture of 7.5 mL ofsuljate E. N,I-dimethyl-4-(methylamino)-IH-imidazole-5-
standardsolution (10 ppm SO.J Rand 7.5 mL of distilled carboxamide (ceffeldine),
water R.
F. I, 7-d.imethyl-3,7-dihydro-IH-putine-2,6-<1ione.
ASSAY _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Dissolve 0.170 g with heating in 5 mL of anhydrous acetic
acid R. Allow to cool, add 10 mL of acetic anhydride Rand
20 mL of toluene R. Titrate with 0.1 M perchloric acid,
determining the end-point potentiometrically (2.2.20).
I mL of 0.1 M perch/oric acid is equivalent to 19.42 mg
Caffeine Hydrate
of CsH lON,02' (Caffeine Monohydrate, Ph. Eur. monograph 0268)
IMPURITIES
present at a sufficient level, be detected by oneor other 0/ the tests
in the monograph. They are limited by the. general tuaptanu
cruerum for other/unspecified impurities and/or by the general
monograph Substances fur pharmaceutical use (2034). It is
there/ore not necessary to identify these imptmties for
demonstration o/compliance. See also 5.10. Control olimpun·ties 212.2 5743-12-4
in substances for pharmaceutical use) A, B, C, D, EJ F.
Action and use
Central nervous system stimulant.
PIlE" _
DEFINITION
1,3,7-Trimethyl-3,7-dibydro-IH-putine-2,6-<1ione
monohydrate.
A. 1,3-dimethyl-3,7-dihydro-IH-putine-2,6-dione
(theophylline), Content
CHARACTERS
Appearance
White or almost white, crystalline powder or silky crystals.
Solubility
Sparingly soluble in water, freely soluble in boiling water,
B. N-(6-amino-l,3-dimethyl-2,4-dioxo-l,2,3,4- slightly soluble in ethanol (96 per cent). It dissolves in
tetrabydropyrimidin-5-yl)formamide, concentrated solutions of alkali benzoates or salicylates.
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1-376 Caffeine Hydrate 2022
It sublimesreadily. Run time 1.5 times the retention time of caffeine.

IDENTIFICATION Identification of impurities Use the chromatogram supplied
First identification: A, B, E. with caffeine for system suitability CRS and the chromatogram
to impurities A, C) D and F.
A. Melting point (2.2.14): 234 'C to 239 'C, determined
Relative mention With reference to caffeine (retention
after drying at 100-105 'C.
time ::::: about 8 min): impurity C == about 0.38,;
B. Infrared absorption spectrophotometry (2.2.24). impurity D == about 0.42; impurity F ::::: about 0.6;
Preparatian Dry the substance to be examined at =
100-105 'C before use. System suitability Reference solution (b):
Comparison caffeine CRS. - resolution: minimum 2.0 between the peaksdue to
C. To 2 mL of a saturated solution add 0.05 mL of iodinated impurities C and D; minimum 2.5 between the peaksdue
potassium iodide solution R; the solutionremains clear. to impurities F and A.
Add 0.1 mL of dilute hydrochloric acidR; a brown precipitate Limits:
is formed. Neutralise with dilute sodium hydroxide solution R; - unspecified impun'ues: for each impurity) not more than
the precipitate dissolves. 0.5 times the area of the principal peak in the
D. In a glass-stoppered tube, dissolve about 10 mg in chromatogram obtainedwith reference solution (a)
0.25 mL ofa mixture of 0.5 mL of acetylacetene Rand 5 mL (0.10 per cent);
of dilute sodium hydroxide solution R. Heat in a water-bath at - total: not more than 0.5 times the area of the principal
80 "C for 7 min. Cool and add 0.5 mL of peak in the chromatogram obtainedwith reference
dimethylaminobenzaldehytk solution R2. Heat again in a water- solution (a) (0.10 per cent);
bath at 80 'C for 7 ntin. Allow to cool and add 10 mL of - disregard limit: 0.25 times the area of the principal peak in
wow R; an intense blue colour develops. the chromatogram obtainedwith reference solution (a)
E. Loss on drying (see Tests). (0.05 per cent).
F. It gives the reaction of xanthines (2.3.1). Sulfates (2.4.13)
Maximum 500 ppm) determined on 15 mL of solution S.
TESTS
Prepare the standard using a mixture of 7.5 mL of sulfate
Solution S
standard solution (10 ppm SO,,) Rand 7.5 mL of distilled
water R.
water R prepared from distilled Water R, cool, and dilute to
50 mL with the same solvent, Loss on drying (2.2.32)
5.0 per cent to 9.0 per cent) determined on 1.000 g by
Appearance of soludon
Solution S is clear (2.2.1) and colourless (2.2.2, MethodII).
Acidity
solution Rt, the solution is green or yellow. Not more than ASSAY
0.2 mL of 0.01 M sodium hydroxide is required to change the Dissolve 0.170 g, previously dried at 100-105 'C, with
colourof the indicator to blue. heating in 5 mL of anhydrous acetic add R. Allow to cool) and
Related substances add 10 mL of acetic anhydride R and 20 mL of UJluene R.
liquid chromatography (2.2.29). Titrate with 0.1 M perchloTic acid, determining the end-point
examined in the mobile phase and dilute to 50.0 mL with I mL of 0.1 M perchlotic acid is equivalent to 19.42 mg
the mobile phase. Dilute 1.0 mL of this solution to 10.0 mL of C.H 1oN. 0 2'
with the mobile phase. IMPURITIES
Reference solution (a) Dilute 2.0 mL of the test solution to Otherdeuaoble impurities (the following substances would, if
100.0 mL with the mobile phase. Dilute 1.0 mL of this present at a sufficient level, be detected by oneor other of the tests
solution to 10.0 mL with the mobile phase. in the monograph. They are limiud by thegeneral acceptance
Reference solution (b) Dissolve 5 mg of caffeine lor system oiuiion for otherlunspecified impurities andlor by thegeneral
suitabilily CRS (containing impurities A, C, D and F) in the monograph Substances for pharmaceutieal use (2034). It is
mobile phase and dilute to 5 mL with the mobile phase. therefore not neussaryto identify these impun·ties for
Dilute 2 mL of this solution to 10 mL with the mobile demonstration of compliance. See also 5.10. Control of impurities
phase. in substances for pharmaceutical use) A, B, C, D) E, F.
Column:
- size: I::::: 0.15 rn, 0 ::::: 4.6 mm,
- ste,ionary phase: end-capped amidohexade<ylsilyl silka gelfor
Mobile phase Mix 20 volumes of 'elrahydrofuran R,
25 volumes of acetonimle Rand 955 volumes of a solution
containing 0.82 gIL of anhydrous sodium acetere R previously
adjusted to pH 4.5 with glacial acetic acidR. A. 1,3-<1imethyl-3,7-dihydro-IH-purine-2,6-dione
(theophylline),
Plo» rare 1.0 mUntin.
Injection 10 ~.
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2022. Calamine 1-377
Practically insoluble in ffJater. It dissolves with effervescence

in hydrochloric acid.
IDENTIFICATION
A. Yields the reactions characteristic of carbonaus,
Appendix VI.
R To 2 g add 5 mL of hydrochloric acid and heat to boiling;
B. N-(6-amino-l,3-dimelhyl-2,4-dioxo-l,2,3,4- if necessary, add hydrochlotic acid drop wise until a bright
tettahydropyrimidin-5-yl)fonnamide, yellow solution is obtained. Cool and add 13.5M ammonia
until the first sign of precipitate (solution A). The solution
yields reaction B characteristic of iron salts) Appendix VI.
Dilute 1 mL of solution A to 5 mL with water; the solution
yields the reaction characteristic of zinc salts, Appendix VI.
TESTS
Calcium
Dissolve 0.50 g in a mixture of IO mL of waterand 2.5 mL
C. 1,3,9-trimethyl-3,9-dihydro-IH-purine-2,6-dione of glacial acetic add and filter. To 0.5 mL of the filtrate add
(isocaffeine), 15 mL of 5Ar ammonia and 2 mL of a 2.5% wlv solution of
ammonium oxalate and allow to stand for 2 minutes.
The solution remains dear.
Soluble barium salts
To the remainder of the filtrate obtained in the test for
Calcium add 2 mL of 1M sulfuric add and allow to stand for
5 minutes. The solution remains dear.
D. 3,7 -dimethyl-3,7-dihydro-lH-purine-2,6-dione Lead
(theobromine), N9t more than 150 ppm when determined by atomic
absorption spearophotometty, Appendix II D J Method II,
measuring at 283.3 om or 217 om and using an air-acetylene
flame. Carefully add 5 g of the substance being examined to
25 mL of hydrochlonc add and allow to stand for 18 hours.
Add 5 mL of nitric acid and sufficient water to produce
200 ml., Use lead standard solution (l00 ppm Ph) suitably
diluted with a 3.5% vlv solution of nitric acid to prepare the
E. N,I-dimethyl-4-(methylamino)-IH-imidazole-5- standard solution.
carboxamide (caffeidine), Chloride
Dissolve 0.15 g in water with the addition of 1 mL of nmu
add, filter and dilute to 30 mL with water. The resulting
solution complies with the limit ust for chlorides, Appendix VII
(0.07%).
Sulfate
Dissolve 0.1 g in water with the addition of 3 mL of 2M
F. 1,7-dimethyl-3,7-dihydro-IH-purine-2,6-dione. hydrochlorU: acid, filter and dilute to 60 mL with water.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEll The resulting solution complies with the limit test for sulfaleS,
Appendix vn (0.6%).
Ethanol-soluble dyes
Shake 1.0 g with 10 mL of ethanol (90%) and filter.
The filtrate is colourless, Appendix N B, Method II.
Calamine
Prepared Calamine Matter Insoluble In hydrochloric acid
Dissolve 1 g in 20 mL of warm 2Mhydrochloric acid and
Action and use filter. The residue,when washed with waterand dried to
Antipruritic. constant weight at 105'=>, weighs not more than 10 mg.
Preparations Water-soluble dyes
Aqueous Calamine Cream Shake 1.0 g with 10 mL of water and filtet. The filtrate is
Calamine Lotion colourless, Appendix N B, Method II.
Calamine Ointment Residue on ignition
Calamine and Coal Tar Ointment 68.0 to 74.0%, when ignited at a temperature not lower than
9000 until, after further ignition, two successive weighings do
DEFINITION not differ by more than 0.2% of the weight of the residue.
Calamine is a basic zinc carbonate suitably coloured with Use I g.
iron(m) oxide.
CHARACTERISTICS
An amorphous, impalpable, pink or reddish brown powder,
the colour depending on the variety and amount of ironrm)
oxide present and the process by which it is incorporated.
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1-378 Calcifediol 2022
Calcifediol Monohydrate *** Referene< solution (d) Dissolve I mg of calcifediol for

*** *** impurity D identification CRS in the mobile phase and dilute
Calcifediol *** to 10 mL with the mobile phase.
(Ph. Bur. monograph 1295) Column:
=
- size: / 0.15 rn, 0 =
4.6 nun;
- stationary phase: end-capped octylsilyl silica gelfor
CH, Mobile phase waterfor chromatography R, methanol R
HO CH
3
(20:80 VW).
• ><,0 F/qUJ rare 1.5 mlJmin.
Injection 50 IJL of the test solution and reference
Run time Twice the retention time of calcifediol.
418.7 63283-36-3 with calcifediol far impurity D identification CRS and the
Action and use identify the peak due to impurity D.
Vitamin D analogue. Relative retention With reference to calcifedioJ (retention
PhEIK _
time.= about II min): pre-calcifediol = about 1.3;
DEFINITION System suitability Reference solution (c):
(3S,5Z,7Ej-9, I0-Secocholesta-5,7,I O(19)-triene-3,25-diol - resolution: minimum 5.0 between the peaks due to
monohydrate. calcifediol and pre-calcifediol.
Content Limits:
98.0 per cent to L02.0 per cent (anhydrous substance). - impuniy D: maximum 0.3 per cent;
A reversible isomerisation to pre-calcifediol takes place in. - unspecified impurides: for each impurity, maximum
solution, depending on temperature and time. The activity is 0.10 per cent;
due to both compounds (see Assay). - total: maximum 0.5 per cent;
- reporting threshold: 0.05 per cent (0.5 rimes the area of the
CHARACTERS
Appearance reference solution (b»; disregard the peak due to
White or almost white crystals. pre-calcifediol.
SolubUlty Water (2.5.32)
Practically insoluble in water, freely soluble in ethanol 3.8 per cent to 5.0 per cent, determined on 10.0 mg by
(96 per cent), soluble in fatty oils. direct introduction of the sample.
It is sensitive to air, heat and light.
ASSAY
IDENTIFICATION Liquid chromatography (2.2.29) as described in the test for
A. Infrared absorption spectrophotometry (2.2.24). related substances with the following modification.
Comparison Ph. Bur. reference spectrum of calcifediol. Injection Test solution and reference solution (a).
B. Examine the chromatograms obtained in the assay. Calculate the percentage content of C27H440Z taking into
Results The principal peak in the chromatogram obtained account the assigned content of calcifediol CRS and, if
with the test solution is similar in retention time and size to necessary) the peak due to pre-ealcifediol.
the principal peak in the chromatogram obtained with STORAGE
Under nitrogen, in an airtight container, protected from light]
TESTS at a temperature of 2 °C to 8°C.
Related substances The contents of an opened container are to be used
Liquid chromatography (2.2.29): use the nonnalisation irrunediateJy.
procedure. Cany Out the test as rapidly as possible, avoiding
exposure 10 actinic fight and air. )
IMPURITIES
Specified impurities D.
Testsolution Dissolve 5.0 mg of the substance to be
examined without heating in the mobile phase and dilute to Otherdetectable impurities (thefollowing substances would, if
50.0 mL with the mobile phase. present at a safficiem level, be detected by oneor other of the tests
in the monograph. They are limited by thegeneral a"epumce
Reference solution (a) Dissolve 5.0 mg of calcifediol CRS
oiunon for otherlunspedfied impurities and/or by thegeneral
without hearing in the mobile phase and dilute to 50.0 mL
monograph Substances far pharmaceutical US< (2034). It is
with the mobile phase. therefore not necessary to identify these impuriu·es f(JT'
Reference solution (b) Dilute 1.0 mL of reference solution (a) demonstration of compliance: See also 5.10. Control of impurities
to 100.0 mL with the mobile phase. Dilute 1.0 mL of this in substances for pharmaceutical use) A, B, C.
Reference solution (c) Heat 2 mL of reference solution (a) in
a water-bath at 80°C under a reflux condenser for 2 hand
cool.
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2022 Calcipotriol 1-379
Preparations
Calcipotriol and Betamethasone Cutaneous Foam
Ca1cipotriol and Betamethasone Gel
Calcipotriol and Betamethasone Ointment
Calcipotriol Cream
Calcipotriol Ointment
A. 9p, 10il-cholesta-5, 7-diene-3p,25-diol, Calcipotriol Scalp Application
PhEur _
DEFINITION
(5Z,7E,22E,24S)-24-Cyclopropyl-9,10-secochola-5,7,10
( 19),22-tetraene-ICl,3p,24-triol.
Content
HO .-
H
A reversible iscmerisarion to pre-caJcipotriol takes place in
B. cholesta-5,7-diene-3p,25-diol, solution, depending on temperature and time. The activity is
due to both compounds.
CHARACfERS
CH, Appearance
HO CH, White or almost white, crystalline powder.
Solubility
(96 per cent), slightly soluble in methylene chloride.
It is sensitive to heat and light.
IDENTIFICATION
C. (3S,6E) -9, 10-secocholesta-5(10),6,8-triene-3,25-diol, Comparison Ph. Eur. reference spectrum of anhydrous
calcipotriol.
B. Loss on drying (see Tests).
TESTS
Carry out the tests for related substances and the assayas rapidly
as possible and protected from actinic light and air.
Related substances
H,C A. Thin-layer chromatography (2.2.27).
Solution A To 1 rnL of triethylamine R add 9 rnL of
. OH chloroform R.
'H
D. (3S,5E, 7E)-9,1 0-secocholesta-5,7, 1O(19)-triene-3,25-diol. in 100 ~L of solution A.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEur Reference solurion (a) To 10 l'1. of the test solution add
990 ~L of solution A.
Reference solurion (b) To 250 ~L of reference solution (a)
add 750 l'1. of solution A.
Calcipotriol Reference solution (c) To 100 ~L of reference solution (a)
add 900 ~L of solution A.
Anhydrous Calcipotriol Reference solution (d) Place 2 mg of the substance to be
(ph. Eur. monograph 2011) examined in a vial and dissolve in 200 f.lL of solution A.
Close me vial and keep it In a water-bath at 60°C for 2 h.
Plare TLC silica gel F2 s< plateR.
Mobile phase 2-merhylpropanol R, methylene chloride R
(20:80 VIV).
Application 10 j.lL of the test solutionand reference
CH,
Drying In air, then at 140°C for 10 min.
HO·- ',OH
Detection Spray the hot plate with an akoholic solution of
H H
sulfuric mid R dry at 140°C for not more than 1 min and
J
112965-21-6
examine in ultraviolet light at 366 nm.
C"R.oO, 412.6
Relative retention With reference to calcipotriol
Acdon and use (RF = about 0.4): impurity G = about 0.4;
Vitamin D analogue.
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1-380 Calcipotriol 2022
impurity H ;;;; about 0.4; pre-calcipotriol = about 0.9; Limits:

impurity A = about 1.2. - impuniy B: not more than 0.5 times the area of the
System suitability Reference solution (d): principal peak in the chromatogram obtained with
- the chromatogram shows a secondary spot due to reference solution (a) (0.5 per cenr):
pre-calcipctriol. - impurities C, D: for each impurity, not more than the
area of the principal peak in the chromatogram
Limits:
- impurity A: any spot due to impurity A is not more
- unspecified impmities: for each impurity, not more than
intense than the spot in me chromatogram obtained
- impun"ties G, H: any spot due to impurity G or His
not more intense than the spot in the chromatogram
obtained with reference solution (b) (0.25 per cent for
solution (a) (2.S per cent};
the sum);
- disregard limit: 0.5 times the area of the principal peak
- unspecified impurilies: any other spot is not more
intense than the spot in the chromatogram obtained
in the chromatogram obtained with reference
solution (b) (O.OS per cent).
with reference solution (c) (0.1 per cent).
B. Liquid chromatography (2.2.29). Loss on drying
Maximum 1.0 per cent, determined on 5 mg by
Solution A Dissolve 1.32 g of ammonium phosphate R in thermogravimetry (2.2.34). Heat to lOS "C at a rate of
water R and dilute to 10.0 mL with the same solvent. 10 °C/min and maintain at 105°C for 60 min.
Solvent mixture Solution A, water R, methanol R
(0.3:29.7:70 VlVflI). ASSAY
Test solutwn (a) Dissolve 2 mg of the substance to be
related substances with the following modification"
the solvent mixture. Injection Test solution (b) and reference solution (d).
Test solution (b) Dissolve 2.00 mg of the substance to be Calculate the percentage content of C27H4.o0J taking into
examined in the solvent mixture and dilute to 20.0 mL with account the assigned content of cakipotriol monohydrate CRS
the solvent mixture. and, if necessary, the peak due to pre-calcipotriol.
Reference solution (a) Dilute 1.0 mL of test solution (a) 10 STORAGE
100.0 mL with the solvent mixture. In an airtight container, protected from light, at -20°C or
Reference solution (b) Dilute 1.0 rnL of reference solution (a) below.
to 10.0 rnL with the solvent mixture. IMPURITIES
Reference solution (c) Dissolve I mg of cakipotriol for system Test A for related substances
suitaMity CRS (containing impurities B, C aod D) in the A, G, H, I.
solvent mixture and dilute to 2.5 mL with the solvent Test B for related substances
mixture. B, C, D, E, F.
Reference solution (d) Dissolve 2.00 mg of calcipomol Specified impurities A, B, C, D, G, H.
monohydrate CRS in the solvent mixture and dilute to
Otherdeeecusble impurities (the following sub'usnces would, if
present at a sufficient level, be detected by oneor other 0/ the tests
Column: in the monograph. They are limited by thegeneral acceptance
=
- size: 1= 0.10 m, 0 4.0 rom; criterion for other/unspecified impUlities andlor ~ thegeneral
- stationary phase: end-capped octadecylsi/yl silica gelfor monograph Subsusnces for phormaceuucol use (2034). It is
chromatography R (3 pm). there/ore not neussary to idenufy these impun"tres for
J1J.obile phase waterfor chromatography R, methanol R demonstration of compliance. See also 5.10. Control of impuniies
(30:70 VIV). in substances for pharmaceutical use) E, F, I.
Deteaion Spectrophotometer at 264 nm.
Injection 20 ilL of test solution (a) and reference
Run time Twice the retention time of calcipouiol.
Identification of impurities Use the cluomatogram supplied
with calcipotriol for system suitabaity CRS and the
CH,
the peaks due to impurities B, C and D. HO-- • OH
Relative retention With reference to calcipotriol (retention H 'H
time = about 13.S min): impurity B = about 0.86;
= =
impurity C about 0.92; impurity D about 1.3. A. (SZ,7E,22E)-24-cyclopropyl-l",3~-dihydroxy-9,1 0-
System suitability Reference solution (c): secochola-5,7,1 O(19),22-tetraen-24-one,
- peak-ro-valley ratio: minimum 1.5, where Hp :::: height
above the baseline of the peak due to impurity C and
=
H; height above the baseline of the lowest point of
the curve separating this peak from the peak due to
calcipotrio1.
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2022 Calcipotriol 1-381
H,C
HO-- "OH
H H
B. (5Z,7 Z,22E,24S)-24-cyclopropyl-9, 1O-secochola-5,7,10 F. (5Z, 7E,22E,24S)-24-cyclopropyl-Ia,3p-bis[[( I, 1-

(l9),22-tetraene-Ia,3p,24-triol «7Z)-caicipotriolJ. dimethylethyl)dimethylsilyl] oxy]-9, 1O-secochcla-S,7, 10
(l9),22-tetraen-24-ol,
H OH
H,c
HO-- '. OH
H H
HO-- .. OH
H H
C. (5E,7E,22E,24S)-24-cyclopropyl-9, I n-secochola-S,7,10
(19).22-tetraene-Ia,3p,24-triol «5E)-<:alcipotriol),
G. 24,24'-oxybis[(5Z,7E,22E,24S)-24-cyclopropyl-9,IO-
secochola-5,7, 1O(19),22-tetraene-1 a,3p-diol],
HO--
H
D. (5Z, 7E,22E,24R)-24-cyclopropyl-9,1O-secochola-5,7,10
( 19),22-tetraene-1a,3p,24-triol (24-epi-calcipotriol),
HO-- -, OH
H H
CH,
HO-· ,.OH
and enanUomer H H
H. (5Z,7E,22E,24R)-24-cyclopropyl-24-[[(5Z.7E,22E,24S)-
24-cyclopropyl-1 a,3 P-dihydroxy-9.1O-secochola-5,7,10
(19),22-tetraen-24-yl]oxy]-9,1o-secochola-5,7,1O(19),22-
HO" .. OH
rerraene-lu.Sjl-diol,
H H
E. rac-(5Z, 7E,24S)-24-cyclopropyl-9,1O-secochola-5,7,10
(19)-triene-1a,3p,24-trio!,
I. (6S,7R,8R,22E,24S)-24-cyclopropyl-6,8:7,19-<licyclo-
9,1O-secochola-5(1O),22-diene-1 a,3p,24-mo! (suprasterol
of calcipotriol).
_ _ _ _ _ _ _ _ _ _ _ _ _~_ _~ PhE"
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1-382 Calcipotriol Monohydrate 2022
Reference solution (d) Place 2 mg of the substance (0 be

Calcipotriol Monohydrate examined in a vial and dissolve in 200 J.1L of solution A.
(Ph. Eur. monograph 2284) Close the vial and keep it in a water bath at 60°C for 2 h.
Plate TLC silica gelF", plate R.
Mobile phase 2-methylpropanol R, methylene chloride R
(20:80 VIV).
Applialtion 10 J.1L of the test solution and reference
,>1,0
Drying In air, then at 140°C for 10 min.
Detection Spray the hot plate with an akohoN, solutUm of
HO-- . OH sulfuric add R, dry at 140 QC for not more than 1 min and
H H examine in ultraviolet light at 366 om.
Relative retention With reference to calcipotriol
430.6 147657-22-5 (R F =. about 0.4): impurity G = about 0.4;
c"I-4.0"H,O
impurity H = about 0.4; pre-calcipotriol = about 0.9;
Action and use =
Vitamin D analogue. System suitability Reference solution (d):
Preparations - the chromatogram shows a secondary spot due (0
Calcipotriol Cream pre-calcfporricl.
Calcipotriol Ointment Limits:
Calcipotriol Scalp Application - impurity A: any spot due to impurity A is not more
intense than the spot in the chromatogram obtained
POE" ~
DEFINITION - impurities G, H: any spot due to impurity G or H is
not more intensethan the spot in the chromatogram
(5Z,7E,22E,24S)-24-Cyclopropyl-9, I0-secochola-5,7,I0
obtainedwith reference solution (b) (0.25 per cent for
(19),22-tetraene-1 a,3~,24-triol monohydrate.
the sum);
Content - unspecified impun·ties: any otherspot is not more
95.5 per cent to 102.0 per eent (anhydrous substance). intense than the spot in the chromatogram obtained
A reversible isomerisation to pre-calcipotriol takes place in with reference solution (c) (0.1 per cent).
solution, dependingon temperature and time. The activity is B. Liquid chromatography (2.2.29).
due to both compounds. Solution A Dissolve 1.32 g of ammonium phosphate R in
CHARACfERS water R and dilute to 10.0 mL with the same solvent.
Appearance Solvent mixture SolutionA, water R, methanol R
White or almost white, crystalline powder. (0.3:29.7:70 VIVIV).
Solubility Testsolution (a) Dissolve 2 mg of the substance (0 be
Practically insoluble in water, freely soluble in ethanol examinedin the solvent mixture and dilute to 5.0 mL with
(96 per cent), slightly soluble in methylene chloride. the solventmixture.
It is sensitive to light. Test solution (b) Dissolve 2.00 mg of the substance to be
IDENTIFICATION examined in the solvent mixtwe and dilute to 20.0 mL with
Comparison Ph. Eur. reference spectrum of cakipotriol Reference solution (a) Dilute 1.0 mL of test solution (a) to
100.0 mL with the solventmixture.
monohydrate.
B. Water (see Tests).
TESTS Reference solUlion (c) Dissolve I mg of calcipotriol for system
Carry out the "'IS for related subS/ances and the asS'!V as rapidly suitability CRS (containing impurities B, C and D) in the
as possible andprotected from actinic light and air. solvent mixture and dilute to 2.5 mL with the solvent
Related substances mixture.
A. Thin-layer chromatography (2.2.27). Reference solution (d) Dissolve 2.00 mg of calcipotrio1
Solution A To 1 mL of triethylamine R add 9 mL of monohydrate CRS in the solvent mixture and dilute to
chloroform R. 20.0 mL with the solventmixture.
Testsolution Dissolve 1 mg of the substance to be examined Column:
in 100 tu. of solution A. - size: 1= 0.10 m, (2) = 4.0 mm;
Reference solution (a) To 10 ~L of the test solution add - stationary phase: end-capped octadecylsilyl silica gelfor
990 ~L of solution A. chromatography R (3 urn).
Reference solution (b) To 250 ~L of reference solution (a) Mobile phase waterfor chromatography R, methanol R
add 750 ~L of solution A. (30:70 VIV).
Reference solution (c) To 100 ~L of reference solution (a) Flow rate 1.0 mUmin.
add 900 tu.of solution A. Detection Spectrophotometer at 264 om.
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2022 Calcipotriol Monohydrate 1-383
Injection 20 IJL of test solution (a) and reference

Run time Twice the retention timeof calcipotriol.
with calcipotriol for system suitabl7ily CRS and the
the peaks due to impurities B, C and D.
Relative retention Withreference to calcipotriol (retention HO·· '. OH
= =
time about 13.5 min): impurity B about 0.86; H H
impurity C = about 0.92; impurity D = about 1.3.
Systemsuitahl1ity Reference solution (c): A. (5Z, 7E,22E)-24-cyclopropyl-la,3p-dihydroxy-9.10-
- peak-w-valley ratio: minimum 1.5, where Hp = height secochola-s.r, 1O(l9),22-tetraen-24-one,
HI) = height above the baseline of the lowestpoint of
the curve separating this peakfrom the peakdue to
calcipotriol.
Limits:
- impun°ty B: not more than 0.5 times the area of the
principalpeak in me chromatogram obtained with
- impurities CJ D: for each impurity, not more than the
area of the principalpeak in the chromatogram
obtained with referencesolution (a) (1.0 per cent),
- unspecified impumies: for each impurity, not more than B. (5Z, 7Z,22E,24S)-24-cyclopropyl-9, I 0-secochola-5, 7,10
the area of the principalpeak in the chromatogram (19),22-tetcaene-la,3p,24-triol «7Z)-calcipotriol),
obtained with referencesolution (b) (0.10 per cent);
- disregard limit: 0.5 times the area of the principal peak
in the chromatogram obtained with reference
Water (2.5.12)
ASSAY
related substances with the following modification. C. (5E,7 E,22E,24S)-24-cyclopropyl-9, 10-secochola-5,7,10
Injection Test solution (b) and reference solution (d). (19),22-tetcaene-la,3p,24-triol «5E)-ealcipotriol),
Calculate the percentage content of C27H.tOOj takinginto
account the assigned content of calcipotriol monohydrate CRS
and, if necessary, the peak due to pre-ealcipotriol.
STORAGE.
In an airtightcontainer, protected from light.
IMPURITIES
Test A for reJated substances
A, G, H, I.
Test B for related substances
B, C, D, E, F.
Specified impurities AJ B, CJ DJ G, H. D. (5Z, 7E,22E,24R)-24-cyclopeopyl-9,1 0-secochola-5,7,10
(19),22-tetraene-1 a,3p,24-trio! (24-epi-calcipotriol),
present at a sufficient levelJ be detected by oneor other of the tests
criterion for otherhmspecified impurities and/or by the general
therefore not necessary to idemify these impurities for
demonstration of compliance. See 'also 5.10. Control of impurities
in substances for pharmaceutical use) E, FJ I.
HO-- -, OH
H H
E. rac-(5Z,7E,24S)-24-cyclopropyl-9,1 0-secochola-5, 7,10

(19)-triene-la,3p,24-triol,
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1-384 Calcitonin (Salmon) 2022
OH
Calcitonin (Salmon)
H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Gty-
I ,
~-~-~r-~-~-~-~-~-~-~n
lli-~-~-~-~-~n-Thr-~-~-~
Thr-Pro-NH 2
3432
F. (5Z, 7 E,22E,24S)-24-cyclopropyl-I~,3/l-bis[[(I, 1-
dimethylethyl)dimethylsilyl] oxy]-9, IO-secochola-5,7, I 0 Action and use
( 19),22-tetraen-24-01, Hormone.
Preparation
Calcitonin (Salmon) Injection
PIlE,; _
DEFINITION
Polypeptide having the structure determined for salmon
calcitonin I. It lowers the calcium concentration in plasmaof
mammals by diminishing the rate of hone resorption. It is
obtained by chemical synthesis or by a method based on
HO'- .. OH recombinant DNA (eDNA) technology. It is available as an
H H acetate.
CH,
Content
90.0 per cent to 105.0 per cent of the peptide
C1ol5H24o'N'1404SS2 (anhydrous and acetic acid-free
substance).
G. 24,24'-oxybis[(5Z,7E,22E,24S)-24-cyclopropyl-9,10- By convention, for the purpose of labelling calcitonin
secochola-5,7,I O(19),2i-tetraene-I~,3/l-dioIJ, (salmon) preparations, 1 mg of calcitonin (salmon)
(C'4,H,.oN..04SS,) is equivalent to 6000 ill of biological
activity.
PRODUCTION
The following requirements apply only to calcitonin (salmon)
produced by a method based on rDNA technology.
Prior to release, the following tests are carried out on each
batch of calcitonin (salmon), unless exemption has been
granted by the competent authority.
Host-cell-derived proteins
The limit is approved by the competent authority.
CH, Host-cell or vector-derived DNA
The limit is approved by the competent authority.
HO·· ',OH
H H CHARACTERS
Appearance
H. (5Z, 7E,22E,24R)-24-cyclopropyl-24-[[(5Z, 7E,22E,24S)- White or almost white powder.
24-cyclopropyl-I~,3P-dibydroxy-9, Io-secochola-5,7,10
Solubility
(19),22-tetraen-24-yl]oxy]-9, I0-secochola-5,7, I0(19),22-
Freely soluble in water,
tetraene-I ct,3P-diol,
IDENTIFICATION
A. Examine the chromatograms obtained in the assay.
reference solution.
The following requirement applies onryto calciumin (salmon)
obtained by chemical synthesis.
B. Amino acid analysis (2.2.56).
Express the content of each amino acid in moles. Calculate
1. (6S,7R,8R,22E,24S)-24-cyclopropyl-6,8:7,19-dicyclo- the relative proportions of the amino acids taking as
9,1 0-secochola-5(l 0),22-diene-1 a,3p,24-triol (suprasterol equivalent to 1 the sum, divided by 20, of the number of
of calcipotriol). moles of aspartic acid, glutamic acid, proline, glycine, valine,
leucine, histidine, arginine and lysine. The values fall within
_____~---------------P"'"
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2022 Calcitonin (Salmon) 1-385
thefollowing limits: aspartic acid: 1.8 to 2.2; glutamic acid: TESTS

2.7to 3.3; proline: 1.7 to 2.3; glycine: 2.7 lO 3.3; valine: Acetic acid (2.5.34)
0.9to 1.1; leucine: 4.5 to 5.3; histidine: 0.9 to 1.1; arginine: 4.0 per cent to 15.0 per cent.
0.9to 1.1; lysine: 1.8 to 2.2; serine: 3.2 to 4.2; threonine: Testsolution Dissolve 10.0 mg of the substance to be
4.2to 5.2; tyrosine: 0.7 to 1.1; half-cystine: 1.4 to 2.1. examined in a mixture of 5 volumes of mobile phase Band
Thefollowing requirement applies only to calcitonin (salmon) 95 volumes of mobile phase A and dilute to 10.0 mL with
produced by a method based on rDNA technology. the same mixture of mobile phases.
C. Peptide mapping (2.2.55). Related substances
SELEC11VE CLEAVAGB OF THE PEPTIDE BONDS Liquid chromatography (2.2.29): use the normalisation
Test solution Prepare a 1 mg/ml; solution of the substance to procedure.
be examined. Transfer 1.0 mL to a clean tube. Add 100 It1. ThefolWwing requirement applies to calcitonin (salmon)J whether
of I M tris-hydroeh/oride buffersolution pH 8.0 Rand 20 ~L of obtained ~ chemical syntheris or ~ a method based on rDNA
a freshly prepared 1.0 mglmL solution of trypsin for peptide t«knology.
mapping R. Allow to stand at 2-8 °C for 16-20 h. Stop the A. Test solution. Prepare a 1.0 mglmL solurlon of the
reaction by adding 10 J.lL of a 50 per cent VIV solution of substance to be examined in mobile phase A.
tnfluoroacetic acid R. Cap the vial and mix. Centrifuge the
Reference solution Dissolve the contents of a vial of calcitonin
vials to remove air bubbles. (salmon) CRS in mobile phase A (0 obtain a concentration of
Reference solution Prepare at the same time and in me same 1.0 mglmL.
manner as for the test solution but using calcitonin
Resolution solution Dissolve the contents of a vial of N-cu:etY/-
(salmon) CRS instead of the substance to be examined. Cysl-calcitonin CRS in 400 ~L of mobile phase A and add
CHROMATOGRAPHIC SEPARATION Liquid chromatography 10~ J1L of the test solution.
(2.2.29).
Column:
Column: - size: 1= 0.25 m, 0 = 4.6 mm;
- size: 1= 0.25 m, 0 ::: 4.6 mm; - stationary phase: oaadecylsilyl silica gelfor
- stationary phase: octadecylsilyl sih'ca gelfor chromatography R (5 um);
chromatography R (5 J-Im) with a pore size of 30 om. - temperature: 65 "C.
Mobile phase: Mobile phase:
- mobile phase A: mix 1 mL of trifiuoroacelu; acidRand - mohr1e phase A: dissolve 3.26 g of tetramethylammonium
1000 mL of water R; filter and degas; hydroxide R in 900 mL of water R, adjust to pH 2.5
- mobile phase B: mix 0.850 mL of trijluoroacetic acid R, with phosphorU: acid R and mix with 100 mL of
200 mL of water Rand 800 mL of acetonitrik for aceton;nlle for chromatography Rj filter and degas;
chromatography R; filter and degas; - mobile phase B: dissolve 1.45 g of tetramethylammonium
hydroxide R in 400 mL of water R, adjust to pH 2.5
Tim. MobUe phase A MobIle phllSe B with phosphoric acid R and mix with 600 mL of
(min) (per cent JIM (per cent JIM
acetonitrile for chromatography R; filter and degas;
0-50 100 --> 65 o --> 35
50 - 60 65 --> 40 35 ..... 60 Time MobUe phase A MobUe phase B
60 - 6O.J 40 ..... 0 60 --> 100 (min) (per cent JIM (per cent VIP)
60.1 • 65.1 0 100 0·30 72 48 28 ..... 52
65.1 - 65.2 0-->100 100 --> 0 30 - 32 48 72 52 --> 28
65.2 - 80.2 100 0 32 - 55 72 28
Flow rale 1.2 mUmin. Flow rate 1.0 mllrnin.

Detection Spectrophotometer at 214 nm. Detection Spectrophotometer at 220 om.
Equilibration At initial conditions for at least 15 min. Cany Injection 20 ~L.
out a blank run using the above-mentioned gradient.
Relative retention With reference to calcitonin (salmon)
Injection 20 ~L. (retention time = about 20 min): impurity B = about 0.8;
System suitability The chromatogram obtained with the impurity C = about 0.9; impurity D = about 1.05j
reference solution is qualitatively similar to the chromatogram impurity A = about 1.15.
of calcitonin (salmon) digest supplied with calcitonin System suitability Resolution solution:
(salmon) CRS. ...:..... resolution: minimum 5.0 between the peaks due to
Results The profile of the chromatogram obtained with the calcitonin (salmon) and impurity A)
test solution corresponds to that of the chromatogram - symmetryfoetor: maximum 2.5 for the peak due to
obtained with the reference solution: the retention times of impurity A.
the fragment peaks in me chromatogram obtained with the Limits:
test solution are within 5 per cent of the retention times of - impurities A, B, C, D: for each impurity, maximum
the fragments obtained with the reference solution; the peak 3.0 per cent; other unidentified, specified impurities
area ratios of the fragment peaks in the chromatogram may occur that co-elute with impurities A) B) C
obtained with the test solution) normalised to the area of and D; the acceptance criterion applies irrespective of
peak T2 ) are within 5 per cent of the corresponding peak whether these impurities co-elute;
ratios in the chromatogram obtained with the reference - total: maximum 5.0 per cent;
solution. - disregard limit: 0.1 per cent.
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1-386 Calcitonin (Salmon) 2022
Thefollowing requirement applies only to calcitonin (salmon) Calculate the content of calcitonin (salmon)
produced by a methodbased on rDNA teehnology. (C145H240N4404SS2) from the area of the principal peak in
B. Test solution. Prepare a 0.5 mglmL solution of the each of the chromatograms obtained with the test solution
substance to be examined. To 1.0 mL of this solution add and the reference solution and the declared content of
100 ur, of 0.25 M curate buffer solution pH 3.0 R. C145H24oNH04SS2 in calcitonin (salmon) CRS. Proceed with
Resolution solution Prepare a 1 mglmL solution of the tangential integration of the peak areas.
substance to be examined. M..ix 1 volume of the solution and STORAGE
1 volume of ealcitonin-Gly CRS. To 1.0 mL of this mixture Protected from light at a temperature between 2 DC and
add 100 ~L of 0.25 M titrate buffer solution pH 3.0 R. 8 DC. If the substance is sterile, store in a sterile, airtight,
Column: tamper-evident container.
- size: I ::;;; 0.20 m, 0 = 4.6 mrn; LABELLING
- stationary phase: a suitable polysulfoethylaspanamide The label ,tates:
ion-exchange gel (5 pm). - the calcitonin peptide content (Cl4jH240N4404SS2);
Mobik phase: - the origin: synthetic or rDNA technology.
~ mobile phaseA: mix 15 volumesof acetonitrile for
IMPURITIES
ehromatography Rand 85 volumes of a 2.72 gIL
Specified impurities A, B, C, D, E, ~ G.
solution of potassium dihydrogen phosphate R adjusted
to pH 5.0 with a 600 gIL solution of potassium
o
-
hydroxide R;
mobile phase B: mix 15 volumes of acetonitrile for
>-- Cys - Set - Asn-leu- set -Thf -Cys- Val-L-leo- Gly-
chromatography Rand 85 volumes of a solution H3C l;s-Leu-ser-Gln-GIU-Leu-H~-LYS-leu-~~

,.
containing 2.72 gIL of potassium dihydrogen ~-~-~-~-fu-Asn-~-~-~-~
ae
phosphate R and 29.22 gIL of sodium chloride R Thr-Pro-NH 2
adjusted to pH 4.6 with a 600 gIL solution of
potassium hydroxide R;
A. acetylcalcitonin (salmon),
Time MobUe phase A .\lobUe phase B
H· Cys - Sec - Asn- Leu- Set - Thr-Cys~ Val- o-teu - Gly-
(mln) (per cent VIJI) (per cent V/l? , ' 1 0
0-10 100 ..... 0 0 ..... 100 ~-~-Ser-~-~-~u-~-~-~-~

,.
10 - 15
15 - 15.1
0
·0 ..... 100
100
100 ..... 0
~-~-~-~-~-Asn-~-~-ser-G~
,.
Thr-Pro-NH 2
15.1-22.[ 100 0
B. [9-o-leucine]calcitonin (salmon),
Detection Spectrophotometer at 220 run. H-Cys -Ssc - Asn-leu· Sec - Thr- cys- Val-Leu- GIy-
, 10
Injection 50 tIl.; rinse the injector with a 40 per cent VIV ~-~u-ser-~-~-~-~-~-~-~
,.
solution of acetonitrile for chromatography R. The-Pro - Arg ~ The -Aso - ltv -Gly -Sec -Gly - Thr-
Relative retention With reference to calcitonin (salmon) Pro-N~ "
=
(retention time about 9 min): impurity G = about 0.4;
impurity F = about 0.6; impurity E = ahout 0.9. c. des-22-tyrosine-calcitonin (salmon),
System sU;uWiJity Resolution solution: D. O-acetyJated calcitonin (salmon),
impurity E and calcitonin (salmon). H-Cys- Set - Asn-Leu - Ser - Thr- Cys - Val-L-Leu-GIy-
, • 10
Limits: ~-~-Ser-~-~-~-~-~-~-~-
- impurity E: maximum 0.6 per cent;
- impurities F, G: for each impurity, maximum Thr Tyr- Pro- Arg- Thr- Asn - Thr -Gly-Ser -GIy-
r- "
0.2 per cent. Thr-prO-~~~H j(l
Water (2.5.32)
Maximum 10.0 per cent. E. salmon calcitoniny]g1ycine,
Acetic acid and water
Maximum 20 per cent, calculated by adding together the ~ S03H
]1 31
percentage contents of acetic acid and water determined by H - Ala - Sec- Asn-leu- Ser- Thr- Ala - Val-leu -Gly ~
the methods described above.
~-~-~-~-~-~-~-~-~-~-
,."
Thr-~-~-~-Thr-Asn-Thr-~-Ser-~-
Less than 25 IU/mg, if intended for use in the manufacture
'"
F. [I,7-bis{3-sulfo-L-alanine)]calcilonin (salmon),
ASSAY
Liquid chtomatography (2.2.29) as described in the test for
related substances. Use method A for calcitonin (salmon)
obtained by chemical synthesis and method B for calcitonin
(salmon) ohtained hy a method based on rDNA technology.
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2022 Calcitriol 1-387
S~H S~H
3' ]1
H - Ala - Ser - Asn-leo - Ser- Thr - Ala - Val-leu - GIy- examined without heating in 10.0 mL of the mobile phase.
~-~-~-~-G~-~-~-~-~U-~- '" Reference solution (a) Dissolve 1.00 mg of calcitrinl CRS
~-~-~-~-~-Mn-~-~-~-~- '" without heating in 10.0 mL of the mobile phase.
H ............... C~H
Thf-Pro-N '" Reference solution (b) Dilute 1.0 mL of reference solution (a)
to 100.0 mL with the mobile phase. Dilute 1.0 mL of this
G. [I,7-bis(3-sulfo-L-a1anine))calcitoninylglycine (sabnon).
Reference solution (c) Heat 2 mL of reference solution (a) at
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
80 °C for 30 min.
Column:
- size: I;;;; 0.25 rn, 0 ;;;; 4.6 mm;
- stationary phase: oetylsi/yl silica gelfur chromatography Rl
Calcitriol ***
*** *** (5 prn);
- temperature: 40 QC.
(Ph. Eur. monograph 0883) .***
Mobile phase Mix 450 volumes of a 1.0 gIL solution of
tris(hydroxymethyl)aminomethane R adjusted to pH 7.0-7.5
with phospboric acid R) and 550 volumes of acetonitrile R.
Injection 50 IlL·
Run time Twice the retention time of calcitriol,
CH, Reladveretention With reference to calcittiol (retention
time ;;;; about 14 min): impurity C ;;;; about 0.4;
HO-- • OH
pre-calcittiol ;;;; about 0.88; impurity A ;;;; about 0.95;
H H impurity B = about 1.1.
System suiuzbility:
416.6 32222-06-3
pre-calcitriol and caleitriol in the chromatogram obtained
Action and use
with reference solution (c);
Vitamin D analogue.
- number of theoretical plates: minimum 10 000, calculated
Preparation for the peak due to calcitriol in the chromatogram
Calcilriol Capsules obtained with reference solution (a).
PhE" _ Limits:
- impurities A~ B) G: for each impurity, maximum
DEFINITION 0.5 per cent;
(5Z,7E)-9,1 0-Secocholesta-5,7,1O(19)-lriene-lex,3p,25-lriol. - unspecified impurities: for each impurity, maximum
Content 0.10 per cent;
97.0 per cent to 103.0 per cent. - total: maximwn 1.0 per cent;
A reversible isomerisation to pre-calcitriol takes place in - disregard limit: 0.5 times the area of the principal peak in
solution, depending on temperature and time. The activity is the chromatogram obtained with reference solution (b)
due to both compounds (see Assay). (0.05 per cent); disregard the peak due to pre-calcitriol,
CHARACTERS ASSAY
Appearance Liquid chromatography (2.2.29) as described in the test for
White or almost white crystals. related substances with the following modifications.
Solubility Injection Test solution and reference solution (a).
PracticaUy insoluble in water, freely soluble in ethanol System SUil~bilitY Reference solution (a):
(96 per cent), soluble in fatty oils. - repeaUlbility: maximum relative standard deviation of
It is sensitive to air, heat and light. 1 per cent for the peak due to calcitriol after 6 injections.
Calculate the percentage content of C27H4403 taking into
IDENllFlCATION
account the assigned content of cakitriol CRS and, if ,
necessary) the peak due to pre-ealcitriol.
Comparison Ph. Eur. reference spectrum of calcr'triol.
STORAGE
Under nitrogen) in an airtight container, protected from light)
Results The principal peak in the chromatogram obtained at a temperature of 2 QC to 8 QC.
The contents of an opened container are to be used
Immediately.
IMPURITIES
TESTS
Related substances
procedure. Carry out the test as rapidly as possible~ avoiding
exposure to actinic light and air.
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1-388 Calcium Acetate 2022
CHARACTERS
Appearance
White or almost white, hygroscopic powder.
Solubility
Freelysoluble in water, slightly soluble in ethanol
(96 per cent).
H,c
IDENTIFICATION
HO-· ,.OH A. It gives reaction (b) of calcium (2.3.1).
H H B. It gives reaction (b) of acetates (2.3.1).
A. (5B,7E)-9 ,I 0-secocholesta-5,7 ,I O(19)-triene-1",3P,25-triol TESTS

(trans-calcitriol), Solution S
Dissolve5.0 g in carbon dioxide-free waterR and dilute [Q
Solution S is clear (2.2.1) and colourless (2.2.2, Method11).
pH (2.2.3)
7.2 to 8.2.
Dilute 5.0 mL of solution S to 10.0 mL with carbon dioxide-
free water R.
Readlly oxidisable substances
Dissolve 2.0 g in boiling water R and dilute to 100 mL with
boiling water R. Add a few glass beads, 6 mL of 5 M ,ulfuric
B. (5Z,7E)-9,1 0-secocholesta-5,7,1 O(19)-triene-1 P,3P,25-triol acid Rand 0.3 mL of a 3.2 gIL solution of potassium
(1Il-calcitriol), ptmranganate R. Mix, boil gentlyfor 5 min and allow the
precipitate to settle. The pink colour In the supernatant is
not completely discharged.
Chlorides (2.4.4)
o Maximum 330 ppm.
>-N Dissolve 0.15 g in water R and dilute to 15 mL with the
O N
o
'uN
I same solvent.
Fluorides
CH, Maximum 50 ppm.
HO·· Potentiometry (2.2.36, Me/hodI).
H Testsolution In a 50 mL volumetric flask, dissolve 0.200 g
in a 10.3 gIL solution of hydrrxhlori< acid R, add 5.0 mL of
C. (6aR,7R,9aR)-II-[(3S,5R)-3,5-dihydroxy-2- jlUQride standard solutiou (1 ppm F) R and dilute to 50.0 mL
methyleyclohex-l-enyl]-7-[(I R)-5-hydroxy-l, 5- with a 10.3 gIL solution of hydrochlatic acidR. To 20.0 mL
dimethylhexyl)-6a-methyl-2-phenyl-5,6,6a,7,8,9,9a,llof the solution add 20.0 mL of total-ionic-strength-adjusunent
octahydro-I H,4aH-eyclopenta If] [I ,2,4] triazolo[ I ,2-aI buffer Rand 3 mL of an 82 gIL solution of anhydrous sodium
cinnoline-I,3(2H)-dione (triazoline adduct of acetate R. Adjust to pH 5.2 with ammonia R and dilute to
pre-calcittiol) . 50.0 mL with distiUed water R.
____________________ 1'/1,,, Reference ,oIuriaus To 0.25 mL, 0.5 mL, 0.75 mL and
1.0 mL ofjluoride standard sdution (10 ppm F) R add
20.0 mL of total-ionic-strength-adjustment buffer R and dilute to
50.0 mL with distilled water R.
Calcium Acetate Indicator electrode Fluoride selective.
Take into account the addition of fluoride to the test solution
for the calculation.
ea"
Nitrates
To 10.0 mL of solution S add 5 mg of sodium chloride R,
158.2 62-54-4 0.05 mL of indigo carmine solution R and add with, stirring,
10 mL of nitrogen-free ,u/furic acid R. The blue colour
Acdon and use remains for at least 10 min.
Used in solutions for haemodialysis and peritoneal dialysis.
Sulfates (2.4.13)
1'/1,,, _ Maximum 600 ppm.
DEFINITION Dissolve 0.25 g in distilled water R and dilute to 15 mL with

the same solvent.
Calcium diacetate.
Content
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2022 Calcium Ascorbate 1-389
Aluminium (2.4.17) Water (2.5.12)

Maximum 1 ppm, if intended for use in the manufacture of Maximum 7.0 per cent, determined on 0.100 g. Add 2 mL
peritoneal dialysis solutions, haemofiltration solutions or of anhydrous acetic acid R to the titration vessel in addition to
haernodialysis solutions. the methanol. Clean the titration vessel after each
Test solution Dissolve 4.0 g of the substance to be examined determination.
in 100 mL of water R and add 10 mL of acelate buffer solution ASSAY
pH6.0R. Dissolve 0.150 g in 100 mL of waterR and carry out the
Reference solution M.ix 2 mL of aluminium standard solution complexometric titration of calcium (2.5.11).
(2 ppm AD R, 10 mL of oak"e blf/fer solution pH 6.0 Rand J mL of 0.1 M sodium edetate is equivalent to 15.82 mg
98 mL of water R. of C,H.CaO,.
Blank solution Mix 10 mL of oarate buffer solurion pH 6.0 R
STORAGE
and 100 mL of water R.
Iron (2.4.9)
Maximum 20 ppm, If intendedfor use in the manufacture of LABELLING
parenteral preparations or haernodialysis solutions. The label states, where applicable, that the substance is
Dilute 5 mL of solution S to 10 mL of water R.
preparations) peritoneal dialysis solutions) haemofiltration
Magnesium solutions or haemodialysis solutions.
Maximum 500 ppm. ~ 'M"
Atomic absorption spectrometry (2.2.21, lWethod JI).
solvent. Calcium Ascorbate
Reference solutions Prepare me reference solutions using
magnesium standardsolurion (0.1 per cem i\JIg) R, diluted as (ph. Eur. monograph 1182)
necessary with water R.
Source Magnesium hollow-cathode lamp.
Potassium
Maximum 500 ppm, if intended for use in the manufacture
of parenteral preparations or haemodialysis solutions.
Atomic emission spectrometry (2.2.22, Muhod Ii). C 12H..CaO,z,2H2 0 426.3 5743-28-2
Test sohuion Dissolve 1.00 g of the substance to be
Action and use
examinedin water R and dilute to 25.0 mL with the same
Excipient.
solvent.
Reference solutions Prepare the reference solutions using PflE" _
potassium standard solution (0.2 per cent K) R, diluted as DEFINITION

necessary with water R.
Calcium di[(R)"2-[(S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-
Wavelength 766.5 om. 2H-furan-3-olate] dihydrate.
Sodium Content
Maximwn 500 ppm, if intended for use in the manufacture 99.0 per cent to 100.5 per cent of CI2H14CaOI2,2H20.
of parenteral preparations or haemodialysis solutions.
CHARACTERS
Atomic emissionspectrometry (2.2.22, Method II).
Appearance
Test solution Dissolve 1.00 g of the substance to be White or slightly yellowish, crystalline powder.
Solubility .
solvent.
Freelysoluble in water) practically insoluble in ethanol
Reference solutions Prepare the reference solutions using (96 per cent).
sodium standardsdution (200 ppm Na) R, diluted as necessary
with water R. IDENTIFICATION
Wavelength 589 om. First identification: A) B) E.
Strontium Second identification: A) C) D) E.
Maximum 500 ppm, if intended for use in the manufacture A. Specific optical rotation (see Tests).
of parenteral preparations or haemodialysis solutions. B. Infrared absorption spectrophotometry (2.2.24).
Atomic emission spectrometry (2.2.22) Method 11). Comparison Ph. Eur. reference spectrum of calcium ascorbate.
Test solution Dissolve 2.00 g of the substance to be C. Dilute I mL of solution S (see Tests) to 10 mL with
examined in water R and dilute to 100.0 mL with the same water R. To 2 mL of the solution add 0.2 mL of a 100 gIL
solvent. solution of ferrous sulfate R. A deep violet colour develops.
Reference sonuions Prepare the reference solutions using D. To I mL of solution S add 0.2 mL of dl,ute nitric acid R
strontium standard solution (1.0 per centSr) R) diluted as and 0.2 mL of siroer nitrate solution R2. A grey precipitate is
necessary with water R. formed.
Wavelength 460.7 om. E. The substance gives reaction (b) of calcium (2.3.1).
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1-390 Calcium Carbonate 2022
TESTS ASSAY
Solution S Dissolve 80.0 mg in a mixture of 10 mL of dilute sulfim"c
Dissolve 5.00 g in carbon dioxide-jree waterR and dilute to acid R and 80 mL of carbon dioxide-free waterR. Add I mL of
50.0 mL with the same solvent. search solution R. Titratewith 0.05 M iodine until a persistent
Appearance of solution violet-blue colouris obtained.
Solution S is clear (2.2.1) and not more intensely coloured I mL of 0.05 M iodine is equivalent to 10.66 mg
than reference solution Y6 (2.2.2.1 Method If). Examine the of CI2HI4CaOI2,2HzO.
colour of the solution immediately after preparation of the STORAGE
solution.
In a non-metallic container, protected from light.
pH (2.2.3) _________ ~ ~PhE"

+ 95 to + 97 (dried substance), determined using fresbly
Calcium Carbonate ***
*** ***
prepared solution S.
Related substances
The thresholds indicated under Related substances (ph. Eur. monograph 0014) ***
(Table 2034.-1) in the general monograph Substances/or CaCO, 100.1 471-34-1
Fluorides Action and use
Maximum 10 ppm. Antacid.
Pctentiometry (2.2.36, Method 1). Preparations
Test soluuon In a 50 mL volumetric flask, dissolve 1.000 g Calcium Carbonate Chewable Tablets
in a 10.3 gIL solution of hydrochloric acid R, add 5.0 mL of Calciwn Carbonate Oral Suspension
fluoride standard solution (/ ppm F) R and dilute to 50.0 mL Calcium and Colecalciferol Tablets
with a 10.3 gIL solution of hydrochloric acid R. To 20.0 mL Calciwn and Colecalciferol Chewable Tablets
of rite solution add 20.0 mL of total-ionic-strength-adjustment
PhE" _
buffer Rand 3 mL of an 82 gIL solution of anhydrous sodium
tuetate R. Adjust to pH 5.2 with ammonia R and dilute to DEFINITION
50.0 mL with distilled water R. Content
Reference solu'ians To 0.2'> mL, 0.5 mL, 1.0 mL, 2.0 mL 98.5 per cent to 100.5 per cent (dried substance).
and 5.0 mL of fluoride standard solution (10 ppm F) R add
20.0 mL of total-ionic-strength-adjustment bufferR and dilute to
CHARACfERS
50.0 mL with distined water R. Appearance
Indicator electrode Fluoride selective.
Reference electrode Silver-silver chloride. Solubility
Practically insoluble in water.
Take into account the addition of fluoride to the test solution
for the calculation. IDENTIFICATION
Copper A. It gives the reaction of carbonates (2.3.1).
Maximum 5 ppm. B. 0.2 mL of solution S (see Tests) gives the reactions of
Atomic absorption spectrometry (2.2.23, Melhod1). calcium (2.3.1).
Test solution Dissolve 2.0 g in a 9.7 gIL solutionof mmc TESTS
add R and dilute to 25.0 mL with the same acid solution. Solution S
Reference solutions Prepare the reference solutions using Dissolve 5.0 g in 80 mL of dilute acetic add R. When the
copper standard solution (10 ppm Cu) R, diluting with a effervescence ceases, boil for 2 min. Allow to cool, dilute to
9.7 gIL solution of ninic acid R. 100 mL with dilute acetic acid R and filter, if necessary,
through a sintered-g1ass filter (2.1.2). Keep the residue for
the test for substances insoluble in acetic acid.
Wavelenglh 324.8 om.
Substances insoluble in acetic acid
Aromisation device Air-acetylene flame. Maximum 0.2 per cenr.
Iron Wash any residueobtainedduringthe preparation of
Maximum 2 ppm. solution S with 4 quantities) each of 5 ml.., of hot warer R
Atomic absorption spectrometry (2.2.23, Metlwd 1). and dry at 100-105 "C for I h. The residue weighs a
Test solution Dissolve 5.0 g in a 9.7 gILsolution of nitn"c maximum of 10 mg.
add R and dilute to 25.0 mL with the same acid solution. Chlorides (2.4.4)
Reference solutions Prepare the reference solutions using iron Maximum 330 ppm.
standard solution (10 ppm Fe) R, diluting with a 9.7 gIL Dilute 3 mL of solution S to 15 mL with water R.
solutionof nitric acidR. Sulfates (2.4.13)
Source Iron hollow-cathode lamp. Maximum 0.25 per cent.
Wavelenglh 248.3 om. Dilute 1.2 mL of solution S to 15 mL with distilled lOoter R.
Aromisation device Air-acetylene flame. Iron (2.4.9)
Loss on drying (2.2.32) Maximum 200 ppm.
Maximum 0.1 per cent, determined on 1.000 g by drying in Dissolve 50 mg in 5 mL of dilute hydrochloric acid Rand
an oven at 105 °C for 2 h. dilute to to mL with waterR.
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2022 Calcium Chloride 1-391
Magnesiwn and alkali metals IDENTIFICATION

Maximum 1.5 per cent. A. Solution S (see Tests) gives reaction (a) of chlorides
Dissolve 1.0 g in 12 mL of dilute hydrochloric acid R. Boil the (2.3.I).
solution for about 2 min and add 20 mL of waler R, 1 g of B.lt gives reaction (b) of calcium (2.3./).
ammonium chloride Rand 0.1 mL of methyl red solution R. C. It complieswith the limits of the assay.
Add diluteammonia Rl until the colour of the indicator
changes and then add 2 mL in excess. Heat to boilingand TESTS
add 50 mL of hot ammonium oxalate solution R. Allow to Solution S
stand for 4 h, dilute to 100 mL with water R and filter Dissolve 10.0 g in carbon dioxide-free water R prepared from
through a suitable filter. To 50 mL of the filtrate add distiUtd wafer R and dilute to 100 mL with the same solvent.
0.25 rnLof sulfw;c acid R. Evaporate (Q dryness on a water- Appearance of solution
bath and ignite to constant mass at 600 ± 50 DC. Solution S is clear (2.2.1) and not more intensely coloured
The residue weighs a maximwn of 7.5 mg. than reference solution Y. (2.2.2, Methad /I).
Loss on drying (2.2.32) Acidity or alkalinity
Maximum 2.0 per cent, determined on 1.000 g by drying in To 10 mL of freshly prepared solution S add 0.1 mL of
an oven at 200 ± 10 DC. phenolphthalein solution R. If the solutionis red) not more
ASSAY than 0.2 mL of 0.0/ M hydrochloric acid is required to
Dissolve 0.150 g in a mixture of 3 mL of dilute hydrochloric discharge the colour and if the solutionis colourless, not
acid Rand 20 mL of water R. Boil for 2 min, allow to cool more than 0.2 mL of 0.01 M sodium hydroxide is required to
and dilute to 50 mL with water R. Carry out the tum it red.
complexometric titration of calcium (2.5.11). Sulfates (2.4./3)
J mL of 0.1 M sodium edetate is equivalent to 10.01 mg Maximum 300 ppm.
ercsco; Dilute 5 mL of solution S to 15 mL with dis.1led water R.
FUNCTIONALITY-RELATED CHARACTERISTICS AlumInIum
This seaion provides in/onnalion on characteristics that are To 10 mL of solution S add 2 mL of ammonium chloride
recognised as being relevant control parameters for one or more solution Rand 1 mL of dilute ammonia R1 and boil the
functions oj the substance when usedas an excipient (see chapter solution. No turbidity or precipitate is formed.
5.15). Someof the characteristics desctibed in theFunctionality- If intended for use in the manufacture of dialysis solutions)
related characteristics section may alsobepresent in the mondasory the above test is replaced by the following test for aluminium
part of the monograph sinee they alsorepresent mandatory quality (2.4./7): maximum I ppm.
criteria. In such cases, a cross-reference to the tests described in the fuscribed salution Dissolve 4 g in 100 mL of water Rand
mandatory part is included in the Funaionality-rdaud add 10 mL of acetate buffer solution pH 6.0 R.
charaaetistics section. Control of the characteristit.s can contribuu Reference solution Mix 2 mL of aluminium standard solution
to the qualityof a medicinal product by improving theconsistency
(2 ppm AI) R, 10 mL of acetate buffer solution pH 6.0 Rand
of the manufacturing process and the perfonnance of the medicinal 98 mL of water R.
product during use. W'here control methods are cited, they are
recognised as being suitable for the purpose~ but other methods can Blank salution Mix 10 mL of acetate buffer solution pH 6.0 R
alsobe used. Wherever results for a particular characteristic are and 100 mL of water R.
reponed, the control method must be indicated. Iron (2.4.9)
Thejollowing charaaeristics m~ be relevant for calcium carbonate Maximum 10 ppm) determined on solution S.
used as fillerin tablets and capsules. Magnesium and alka1i metals
Particle-size distribution (2.9.3/ or 2.9.38) Maximum 0.5 per cent.
Powder flow (2.9.36) To a mixture of 20 mL of solution Sand 80 mL of waterR
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1""<1 add 2 g of ammonium chloride Rand 2 mL of dilute
ammonia Rt, heat to boilingand pour into the boiling
solution a hot solution of 5 g of ammonium oxalate R in
15 mL of waterR. Allow to stand for 4 h, dilute to 200 mL
Calcium Chloride Dihydrate ****• with waterR and filter through a suitable filter. To 100 ml;'
**••*•* of the filtrate add 0.5 mL of mlfuricacid R. Evaporate to
(ph. Eur. monograph 00/5) dryness on a water-bath and ignite to constant mass at
CaCI,,2H,O 141.0 /OOJ5-()4..8 600 ± 50 °C. The residue weighs a maximum of 5 mg.
Preparations ASSAY
Calcium Chloride Injection Dissolve 0.280 g in 100 mL of waterRand carry out the
Compound Sodium Lactate Infusion complexometric titration of calcium (2.5. / I).
PhE<I _ 1 mL of 0.1 M sodium edetate is equivalent to 14.70 mg
of CaCI,,2H,O.
DEFINITION
Content LABELLING
91.0 per cent to 103.0 per cent of CaCI,,2H,O. The label states, where applicable, that the substance is
suitable for use in the manufacture of dialysis solutions.
CHARACTERS
Appearance STORAGE
White or almost white, crystalline powder, hygroscopic. In an airtight container.
Solubility -~ PhE<I
Freely soluble in water, soluble in ethanol (96 per cent).
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1-392 Calcium Chloride 2022
To a mixture of 20 mL of solution Sand 80 mL of water R

Calcium Chloride Hexahydrate add 2 g of ammonium chloride Rand 2 mL of dilute
(ph. Eur. monograph 0707) ammonia Rl, heat to boiling and pour into the boiling
solution a hot solution of 5 g of ammonium oxalate R in
CaCI2>6H,O 219.1 7774-34-7 75 mL of water R. Allow to stand for 4 h, dilute to 200 mL
PhE" _ with water R and filter through a suitable filter. To 100 mL
of the filtrate add 0.5 mL of '"/furi< acid R. Evaporate to
DEFINITION
dryness on a water-bath and ignite to constant mass at
Content 600 ± 50 °C. The residue weighs a maximum of 5 mg.
97.0 per cent to 103.0 per cent of CaCh,6H2 0 .
ASSAY
CHARACTERS
Dissolve 0.200 g in 100 mL of water R. Carry out the
Appearance complexometric titration of calcium (2.5.11).
White or almost white, crystalline mass or colourless crystals.
1 mL of 0.1 M sodium edetate is equivalent to ;!1.91 mg
Solubility of CaCl z,6H,O.
Very soluble in water, freely soluble in ethanol (96 per cent).
LABELLING
It solidifies at about 29 °C.
IDENTIFICATION suitable for use in the manufacture of dialysis solutions.
A. Solution S (see Tests) gives reaction (a) of chlorides _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
(2.3.1).
B. It gives the reactions of calcium (2.3.1).
C. It complieswith the limits of the assay.
TESTS Calcium Dobesilate Monohydrate
Solution S
Dissolve 15.0 g in carbon dioxide-free water R prepared from (ph. Eur. monograph 1183)
distilkd water R and dilute [0 100 mL with the same solvent.
than reference solution Y. (2.2.2, Method II).
c," 0'(XS
[H I O
']
"", oH
H,o
To 10 mL of freshly prepared solution S add 0.1 mL of CI,HIOCaOU':'z,H,O 436.4 ZOIZ3-80-2
phenolphthalein solution R. If the solutionis red, not more PhE" ~ _
than 0.2 mL of 0.01 M hydrochlori< add is required to DEFINITION
discharge the colour and if the solutionis colourless, not
Calciumdi(2,S-dihydroxybenzenesulfonate) monohydrate.
more than 0.2 mL of 0.01 M sodium hydroxide is required to
turn it red. Content
Sulfates (2.4.13)
Maximum 200 ppm. CHARACTERS
Dilute 5 mL of solution S 10 15 mL with distilled water R. Appearance
White or ahnost white, hygroscopic powder.
Aluminium
To 10 mL of solution S add 2 mL of ammonium chloride Solubility
solution Rand 1 mL of dilute ammonia RJ. Heat (Q boiling. Very soluble in water, freely soluble in anhydrous ethanol,
No turbidity or precipitate is formed. very slightly soluble in 2-propanol, practically insoluble in
methylene chloride.
If intended for use in the manufacture of dialysis solutions,
the above test is replaced by the following test for aluminium IDENTIFICATION
(2.4.17): maximum I ppm. A. Ultraviolet and visible absorption spectrophotometry
Prescribed sotution Dissolve 6 g in 100 mL of waterRand (2.2.25).
add 10 mL of acetate buffer solution pH 6.0 R. Test solution Dissolve 0.100 g in water R and dilute to
Reference solution Mix 2 mL of aluminium standard solution 200.0 mL with the same solvent. Dilute 5.0 mL of this
(l ppm AQ R, 10 mL of acetate blfffer solutien pH 6.0 Rand solution to 100.0 mL with water R.
98 mL of water R. Spectral range 210-350 urn.
Blank solution Mix 10 mL of acetate blfffer solution pH 6.0 R Absorption maxima At 221 urn and 301 urn.
and 100 mL of water R. Specific absorbance at the absorption maximum at 301 nm
Barium 174 to 181.
To 10 mL of solution S add I mL of calcium su/fate B. Mix I mL offerric chloridesolution R2, I mL of a freshly
solution R. Afterat least 15 min, any opalescence in the prepared 10 gIL solution of potassium fenicyonide Rand
solution is not more intense than that in a mixture of 1 mL 0.1 mL of nitric acid R. To this mixture add 5 mL of freshly
of distilkd water R and 10 mL of solution S. prepared solution S (see Tests): a blue colour and a
Iron (2.4.9) precipitate are immediately produced.
Maximum 7 ppm, determined on solutionS. C. 2 mL of freshly prepared solution S gives reaction (b) of
Magnesium and alkali metals calcium (2.3.1).
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2022 Calcium Folinate 1-393
TESTS 1 mL of 0.1 1\-1 cerium sulfate is equivalent to 10.45 mg of

Solution S CI2HIOCaOlOS2.
Dissolve 10.0 g in carbon dioxide-free water R and dilute to STORAGE
100 mL with me same solvent.
Solution S, when freshly prepared) is clear (2.2.1) and
IMPURITIES
colourless (2.2.2, Method If). Specified impurities A.
pH (2.2.3)
HOY"')
Related substances ~OH
Liquid chromatography (2.2.29). K£ep aU solutions at 2-8 "C.
Buffer solution Dissolve 1.2 g of anhydrous sodium dihydrogen A. benzene-I,4-diol (hydsoquinone).
phosphate R in 900 mL of waterfor chromaUJgraphy R, adjust _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
to pH 6.5 with disodium hydrogen phosphate solution R and
dilute to 1000 mL wiili waterfor chromatography R.
Calcium Folinate Hydrate ***
** **
examined in waler R and dilute to 10.0 mL with me same
solvent.
Calcium Folinate *****
Reference solution (a) Dilute 1.0 mL of the test solution [0
100.0 mL with water R. Dilute 1.0 mL of this solution 10 (ph. Eur. monograph 0978)
examined and 10 mg of hydroquinone R (impurity A) in
water R and dilute £0 10 mL with the same solvent Dilute
1 mL of this solution to 100 mL with water R.
Gdumn:
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: spherical end-capped octadecylsi/yl silica gel
for chromaUJgraphy R (5 urn). CwH2ICaN,O,,xH,O 511.5 2060570-47-8
Mobile phase acetonirri!e Rl, buffer solution (10:90 VIV). (anhydroussubstance)
Flow rate 0.8 mUmin. Action and use
Deuaion Spectrophotometer at 220 nm. Antidote to folic acid antagonists.
Injection I0 ~L. Preparations
RUtl time 2.5 times me retention time of dobesilate. Calcium Polinate Injection
Relative retention With reference to dobesilate (retention Calciwn FolinateTablets
time = about 6 min): impurity A = about 1.7. PhE<l ~~ _
System suilabiJ,iy Reference solution (b):
- resolution: minimum 8.0 between the peaksdue to DEFINITION
dobesilate and impurity A. Calcium (2S)-2-[4-[[[(6RS)-2-amino-5-fonnyl-4-oxo-
Limits: 1,4,5,6,7,8-hexahydropteridin-6-yl]methyl] amino I benzamidoI
- correction factor. for the calculation of content, multiply the pentanedioate hydrate.
peak area of impurity A by 0.6; Content
- ;mpun·ty A: not more than the area of the principal peak - calciumfolinate (C,oH2ICaN,O,): 97.0 per cent 10
in the chromatogram obtainedwith reference solution (a) 102.0 per cent (anhydrous substance);
(0.1 per cent); - calcium (Ca; A, 40.08): 7.54 per cent to 8.14 per cent
- unspecified impurities: for each impurity, not more than the (anhydsous substance).
area of the principal peak in the chromatogram obtained It contains a variable quantity of water.
- total: not more than twice the area of the principal peak in CHARACTERS
~the chromatogram obtained with reference solution (a) Appearance
(0.2 per cent); White or light yellow, amorphous or crystalline, hygroscopic
- disregard It"mit. 0.5 times the area of the principal peak in powder.
the chromatogram obtained with reference solution (a) Solubility
(0.05 per cent). Sparingly soluble in water, practically insoluble in acetone
Iron (2.4.9) and in ethanol (96 per cent).
Maximum 10 ppm, determined on 10 mL of solution S. The amorphous fonn may produce supersaturated solutions
Water (2.5.12) in water.
4.0 per cent to 6.0 per cent, determined on 0.500 g. It shows polymorphism (5.9).
ASSAY IDENTIFICATION
Dissolve 0.200 g in a mixture of 10 mL of water Rand First identification: A, B, D.
40 mL of dilute sulfuric acidR. Tittate with 0.1 M cerium Second identification: A, C, D.
sulfate, determining the end-point potentiometrically (2.2.211). A. Specific opticalrotation (see Tests).
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1-394 Calcium FoJinate 2022
B. Infrared absorption spectrophotometry (2.2.24). Static head-space conditions that may be used:
Comparison calcium latinate CRS. - equilibration temperature: 80°C;
If the spectra obtained show differences, dissolve the - equilibration time: 20 min;
substance {O be examined and the reference substance - pressurisation time: 30 s.
separately in the minimum volume of water R and add Temperature:
dropwise sufficient acetone R to produce a precipitate. Allow
to stand for 15 min, collect the precipitate by centrifugation, Time Temperature
wash the precipitate with 2 small quantities of acetone Rand (mlD) CCJ
dry. Record new spectra using the residues. Column 125 ..... 185
185
Injection pon 250
Test solution Dissolve 15 mg of the substance to be Detector 250
examined in a 3 per cent VJV solution of ammonia Rand
Dnection Flame ionisation.
Reference solution Dissolve 15 mg of calcium folinate CRS in
Injection A[ least 3 times.
a 3 per cent VIV solutionof ammonia R and dilute to 5 mL
with the same solvent. Limit:
- ethanol: maximum 3.0 per cent.
Plate cellulose for chromatography F2S4 R as the coaring
substance. Related substances
Mobile phase The lower layer of a mixture of 1 volume of Liquid chromatography (2.2.29). Prepare the solutions
isoamyl alcohol R and 10 volumes of a 50 gIL solution of citric immediately be/ore use.
acid monohydrate R previously adjusted to pH 8 with Testsolution Dissolve 10.0 mg of the substance to be
ammonia R. examined in waterR and dilute to 10.0 mL with the same
Application 5 ~L. solvent.
Development Over 213 of the plate. Reference rolurion (a) Dissolve 10.0 mg of calcium
/olinate CRS in waterR and dilute to 10.0 rnL with the same
Drying In air.
solvent.
Deteaion Examine in ultraviolet light at 254 run. Reference solution (b) Dilute 1.0 rnL of the test solution to
Results The principal spot in the chromatogram obtained 100.0 rnL with waterR. Dilute 1.0 rnL of this solution to
with the test solution is similar in position and size to the 10.0 rnL with water R.
principal spot in me chromatogram obtained with the
Reference solution (c) Dissolve 5 mg of calcium folinate for
reference solution.
system suitab.7ity CRS (containing impurities A, E and F) in
D. It gives reaction (b) of calcium (2.3.1), 5 mL of water R.
TESTS Column:
Cany out the tests as rapidly as possible, protededfrom aclinic - size: 1= 0.25 m, 0 = 4 mm,
lighe - stationary phase: end-<apped OClode<y/si1y1 silica gelfor
Solution S chromatography R (5 um);
Dissolve 1.25 g in carbon dioxide-free waterR, heating at - tempera..re: 40°C.
40 °C if necessary, and dilute to 50.0 mL with the same Mobile phase Mix 165 rnL of methanol R and 835 rnL of a
solvent. solution containing 4.0 mL of tetrabutylammonium dihydrogen
Appearance of solution phosphate solution Rand 1.42 g of disodium hydrogen phosphate
Solution S is clear (2.2.1). dihydrate R in waterfor chromatography R, previously adjusted
to pH 7.7 with phosphoric acidR or dilute sodium hydroxide
pH (2.2.3) sduuon R.
6.8 (0 8.0 for solution S.
+ 14.4 to + 18.0 (arthydrous substance), determined on
solutionS. Injun·on 10 !Jl. of the test solution and reference
Absorbance (2.2.25)
Maximum 0.60, determined at 420 om on solution S.
Run time 3.5 times the retention time of folinic acid.
Ethanol
with calcium folinate for system suitability CRS and the
Head-space gas cbromatography (2.2.28): use the standard
additions method.
the peaks due to impurities A, E and F.
Tesl solution Dissolve 0.25 g of the substance to be
Relative retention With reference to folinic acid (retention
examined in water R and dilute (Q 10.0 mL with the same
time = about 13.9 min): impurity E = abour 0.4;
solvent.
Reference solution Dilute 0.750 g of anhydrous ethanol R ro
= =
impurity A about 0.6; impurity F about 0.7.
System suitability Reference solution (e):
1000.0 rnL with waterR.
Column: impurities A and F.
- size: I;;;; 10 m, 0 ;;;; 0.32 rnm;
- correction foaors; multiply the peakareas of the following
- stationary phase: styrene-diviny/benzene copolymer R.
Comer gas nitrogen for chromatography R. impurity A =0.6; impurity E =0.6; impurity F =0.6;
Flow rate 4 mllrnin.
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2022 Calcium Folinate 1-395
- for each impurity, use the concentration of calcium

folinate hydrate in reference solution (b).
Limits:
- impurities A, EJ F: for each impurity, maximum
0.3 per cenq
- unspeaJied impurities: for each impurity, maximum A. (2S)-2-(4-aminobenzamido)pentanedioic acid,
0.20 per cent;
Chloride.
Dissolve 0.300 g in 50 mL of waterR heating at 40°C if B. (2S)-2-[4-[[[(6RS)-2-amino-5-formyl-4-oxo-I,4,5,6,7,8-
hexahydropteridin-6-yl]methyIJ(formyl)amino]benzamido]
necessary. Add 10 mL of dilute nitric acid R and titrate with
0.005 kI silver nitrate determining the end-point pentanedioic acid (5,10-difonnyltetrahydrofolic acid),
1 mL of 0.005 M silver nitrate is equivalent to 0.177 mg of
Cl.
Water
(2.5.12): 10.0 per cent [0 17.0 per cent.
Dissolve 0.100 g in a mixture of 15 mL ofJormamide Rand
50 mL of the titration solvent. Stir for about 6 min before
titrating and use a suitable titrant that does not contain C. (2S)-2-[4-[[(2-amino-4-oxo-I,4-<1ihydrop<eridin-6-yl)
pyridine. methyl]amino]benzamido]pentanedioic acid (folic acid),
ASSAY
Carry out theassays as rapidly as possible) protected from aclinic
light
Calcium
Dissolve 0.400 g in 150 mL of water R and dilute [0 300 mL
with the same solvent. Carry out the complexometric
titratlon of calcium (2.5.11).
1 mL of 0.1 J\1 sodium edetate is equivalent to 4.008 mg of
D. (2S)-2-[4-[[(2-amino-4-oxo-I,4-dihydropteridin-6-yl)
Ca.
methyl](formyl)amino]benzamido]pen,anedioic acid (10-
Calcium folinate fonnylfolic acid»
Liquid chromatography (2.2.29) as described in the <esc for
o ?HO r'YCQ,H
lnj«tion Test solution and reference solution (a).
Calculate the percentage content of C2oH21CaN707 taking N~N');;'''N~
.Jl.. Jl ) H H .
and enantiomer
into account the assigned content of calcium folinate CRS.
~N N N
H H
STORAGE
In an airtight container) protected from light. IT the substance
is sterile) the container is also sterileand tamper-evident. E. 4-[[[(6RS)-2-amino-5-formyl-4-oxo-1 ,4,5,6,7,8-
hexahydrop<eridin-6-yl]methyl]amino]benzoic acid
LABELLING (5-formyl<ettahydrop<eroic acid),
preparations.
IMPURITIES
Specified impurities A, E, F.
Otherdetectable impurities (the following substanus would, if
present at a sufficient level) be detected by one or ocher of the tests
in 'he monograph. They arelimited by thegeneral acceptance
oiunon for otherhmspecified impurities. 1, is therefore not F. (2S)-2-[4- [[(2-amino-4-oxo-I,4,7,8-tettahydropteridin-6-
necessary to identify these impurities for demonstration of yl)methyl](formyl)amino]benzamido]pen[anedioic acid
compliance. See also 5.10. Control of impurities in substances for (IO-formyldihydrofolic acid),
pharmaceutical use) B, C, D, G, 1.
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1-396 Calcium Glucoheptonate 2022
Plate cellulose for chromatography R I as the coating

substance.
jWobile phase anhydrous formic acidR, waterR, acetone R,
butanolR (20:20:30:30 VIVIVIII); use a freshly prepared
mixture.
Application I 0 ~L as bands of 20 mm by 2 mm.
G. (2SJ-2-[4-[[(2-amino-4-oxo-I,4,7,8-tetrahydropteridin-6- Development In a tank previously allowed to saturate for
yl)methyI)amino)benzamido]pentanedioic acid 10 min, over a path of J2 em.
(dihydrofolic acid), Drying In air.
Detection Spray with 0.02 M potassium permanganate.
o H CO:!H
o ~~~co,H - the chromatogram shows 2 clearly separated spots.
H2N-----{
HN
N~ ~
r---V N
- ...
N
aooepimeratC'
HNJ\H
solution (a).
I. (2SJ-2-[ 4-[(6aRSJ-3-amino-l-oxo-I,2,5,6,6a,7- B. 0.2 mL of solution S (see Tests) gives reaction (b) of
hexahydroimidazo[I,5-j]pteridin-8(9H)-yl]benzamido] calcium (2.3.1).
pentanedioic acid «6aRSJ-5,10-methylenetetrahydrofolic TESTS
acid). Solution S
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'" Dissolve 10.0 g in carbon dioxide-free water R prepared from
distilled water R and dilute to 100 mL with the same solvent.
Calcium Glucoheptonate than reference solution Y6 (2.2.2, Method II).
pH (2.2.3)
(Ph. Eur. monograph 1399) 6.0 to 8.0 for solution S.
co,] Reducing sugars
~
HO H •
'HO . ~ Maximum I per cent, expressed as glucose.
ca2. H. .. . . OH andepimer at C· Dissolve 1.0 g in 5 mL of water R with the aid of gentle heal.
HO' ', H Cool and add 20 mL of cupri..cirric solution R and a few glass
[ HO H
HO 2
beads. Heat so that boiling begins after 4 min and maintain
boiling for 3 min. Cool rapidly and add 100 mL of a
2.4 per cent VIVsolutionof glacial acetic add Rand 20.0 mL
490.4 of 0.025 M iodine. With continuous shaking, add 25 mL of a
mixture of 6 volumes of hydrochlori< acidR and 94 volumes
Action and use
of water R until me precipitate dissolves, titrate the excess of
Used in treatment of calcium deficiency.
iodine with 0.05 M sodium thiosulfate using 1 mL of starch
PhE", _ solution R added towards the end of the titration, as indicator.
Not less than 12.6 mL of 0.05 M sodium thiosulfate is
DEFINITION required.
Mixture in variable proportions, of calcium di(D-gburo-D-
gulo-heptonate) and calcium di(o-glycero-D-Uio-heplOnate). Cyanide
Dissolve 5.0 g in 50 mL of water R and add 2.0 g of tartaric
Content acid R. Place this solution in a distillation apparatus (2.2.11).
98.0 per cent. to 102.0 per cent of calcium 2,3,4,5,6,7- The plain bend adapter attached to the end of the condenser
hexahydroxyheptanoate (dried substance). has a vertical part that is long enough to extend to 1 ern
CHARACTERS from the bottom of a 50 mL test-tube used as a receiver.
Appearance Place 10 mL of water Rand 2 mL of 0.1 M sodium hydroxide
White or very slightly yellow, amorphous powder, into the receiver. Distil, collect 25 mL of distillate and dilute
hygroscopic. to 50 mL with water R. To 25 mL of this solution add
Solubility 25 mg of ferrous sulfate R and boil for a short time. After
Very soluble in water, practically insoluble in acetone and in cooling to about 70°C add 10 mL of hydrochloric acid Rt,
ethanol (96 per cent). After30 min, filter the solution and washthe filter. A yeUow
spot appears on the filter; there is no blue or green spot.
IDENTIFICATION
Chlorides (2.4.4)
Maximum 100 ppm.
Test solution Dissolve 20 mg of the substance to he To 5 mL of solution S, add 10 mL of water R.
Sulfates (2.4.13)
Reference solution (a) Dissolve 20 mg of cakium
glucohepwnate CRS in I mL of waterR.
Iron (2.4.9)
Reference solution (b) Dissolve 10 mg of calcium
gluconate CRS in 0.5 mL of the test solutionand dilute to Maximum 40 ppm.
1 mL with waterR. Dilute 2.5 mL of solution S to 10 mL with water R.
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2022 Calcium Gluconate 1-397
Loss on drying (1.1.31) Plate TLC silica gel plare R (5-40 urn) [or TLC silica gel
Maximum 5.0 per cent, determined on 1.000 g by drying in plare R (2-10 11m)].
an oven at 105 °C for 3 h. Mobl1e phase concentrated ammoniaR, ethyl acetate R,
Bacterial endotoxlns (1.6. 1if) water R, ethanol (96 per ceno R (10:10:30:50 VIVIVIV).
Less than 167 IV/g, if intended for use in the manufacture of Application I ~L.
parenteral preparations without a further appropriate Development Over 2/3 of the plate.
Drying At 100°C for 20 min; allow to cool.
ASSAY Detection Spray with a solution containing 10 gIL of cerium
Dissolve 0.800 g in a mixture of 2 mL of 3 M hydrochloric sulfate Rand 25 rJL of ammonium molybdare R in dilute
acid and 150 mL of water R. While stirring, add 12.5 mL of sulfun'c acidR and heat at 105 'C for about 10 min.
0.1 M sodium edeuue, 15 mL of 1 M sodium hydroxide and
Results After 5 min, the principal spot in the chromatogram
0.3 g of hydrox;ynaphthol blue, sodium salt R. Titrate with
obtained with the test solution is similar in position, colour
0.1 AtI sodium edetate until the colour changes from violet to
pure blue.
with me reference solution.
I mL of 0.1 M sodium edeuue is equivalent to 49.04 mg
B. Solution S (see Tests) gives the reactions of calcium
of CI4H26CaOI6' (1.3.1).
STORAGE
TESTS
Solution S
Dissolve 1.0 g in water R heated to 60°C and dilute to
--------------------,""" 50 mL with the same solvent.
At 60°C, solution S is not more intensely coloured than
reference solution Y6 (2.2.2, Melhod II). After cooling, it is
Calcium Gluconate not more opalescent than reference suspension II (2.2.1).
(Ph. Eur. monograph 0171) Organic impurities and boric acid
Introduce 0.5 g into a porcelain dish previously rinsed with
sulfuric acid R and placed in a bath of iced water. Add 2 mL
HHO ..H
•
CO']
"OH
H,O
of cooled sulfuric add R and mix. No yellow or brown colour
develops. Add I mL of chromorrope II B solution R. A violet
Ca" Ho1XH
[ colour develops and does not become dark blue.
HO a The solution is not more intensely coloured than that of a
mixture of I mL of chromorrope II B solution R and 2 mL of
448.4 18016-14-5 cooled su{fitrK acid R.
Sucrose and reducing sugars
\ Action and use Dissolve 0.5 g in a mixture of 2 mL of hydrochloric acid Rl
Used in treatment of calcium deficiency. and 10 mL of water R. Boil for 5 min, aUow to cool, add
Preparations 10 mL of sodium carbonate solution R and allow to stand.
Calcium Gluconate Tablets Dilute to 25 mL with water R and filter. To 5 mL of the
Calcium Gluconate Chewable Tablets filtrate add 2 mL of cupri-tartaric solution R and boil for
Calcium Gluconate Effervescent Tablets I min. Allow to stand for 2 min. No red precipitate is
formed.
PIlE" _
Chlorides (2.4. if)
DEFINITION Maximum 200 ppm.
Calcium bis[(2R,3S,4R,5R)-2,3,4,5,6- Dilute 12.5 mL of solution S to 15 mL with water R.
pentahydroxyhexanoate] monohydrate (calcium di Sulfates (1.4.13)
(n-gluconate) monohydrate), Maximum 100 ppm.
Content Dissolve 10.0 g with heating in a mixture of 10 mL of aceric
98.5 per cent to 102.0 per cent of C12H22Ca014,H20. acid Rand 90 mL of distilled warer R.
CHARACTERS Magnesium and alkali metals
Appearance Maximum 0.4 per cent.
White or almost white, crystalline or granular powder. Dissolve 1.00 g in 100 mL of boiling water R, add 10 mL of
SolublIlty ammonium chloride solution R, 1 mL of ammonia Rand,
Sparingly soluble in water, freely soluble in boiling water. dropwise, 50 mL of hot ammonium oxalate solution R. Allow
to stand for 4 h, dilute to 200 mL with water R and filter.
IDENTIFICATION
Evaporate 100 mL of the filtrate to dryness and ignite.
A. Thin-layer chromatography (2.1.17). The residue weighs a maximum of 2 mg.
Testsolution Dissolve 20 mg of the substance [Q be
examined in 1 mL of water R, heating if necessary in a water-
TA1\1C: acceptance criterion 10' CFU/g (1.6.12).
bath at 60 'C.
Reference solution Dissolve 20 mg of calcium g/uconaee CRS
in 1 mL of water R, heating if necessary in a water-bath at
60 'C.
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1-398 Calcium Gluconate 2022
ASSAY TESTS
Dissolve 0.8000 g in 20 mL of hot water R, allow to cool and Solution S
dilute to 300 mL with wale/" R. Carry out the Dissolve 1.0 g in water R heated to 60 DC and dilute to
complexometric titration of calcium (2.5.11). 50 mL with the same solvent.
1 mL of 0.1 M sodium edetate is equivalent to 44.84 mg Appearance of solution
of CI2HnCaOI4.lHzO. At 60 DC, solution S is not more intensely coloured than
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r reference solution Y6 (2.2.2, Method II). Aftercooling) it is
not more opalescent than reference suspension II (2.2.1).
Organic impurities and boric acid
Place 0.5 g in a porcelain dish previously rinsed with sulfune
Anhydrous Calcium Gluconate acid R and placed in a bath of iced water. Add 2 mL of
cooled sulfuric add R and mix. No yellow or brown colour
(ph. Eur. monograph 2364) develops. Add I mL of chromouope II B solution R. A violet
colour develops and does not become dark blue. Compare
the colour obtainedwith that of a mixture of I mL of
HHO / Hco.'] chromotrope II B sduuon Rand 2 mL of cooled sulfuric acid R.
Ca" Ho1XH
[HO
, ··OH
2 Dissolve 0.5 g in a mixture of 2 mL of hydrochloric acid Rl
and 10 mLof water R. Boil for 5 min, allow to cool, add
10 mL of sodium carbonate solution R and allow to stand for
ClzHnCaOl4 430.4 10 min. Dilute to 25 mL with water R and filter. To 5 mL of
the filtrate add 2 mL of cupti-tartanc solution R and boil for
Action and use I min. Allow to stand for 2 min. No red precipitate is
Used in treatment of calcium deficiency. formed.
PhE<r _ Chlorides (2.4. 4)
Maximum 200 ppm.
DEFINITION
Anhydrous calcium D-g1uconate. Dilute 12.5 mL of solutionS to 15 mL with water R.
Content Sulfates (2.4.13)
98.0 per cent to 102.0 per cent (dried substance). Maximum 100 ppm.
Dissolve 10.0 g with heating in a mixture of 10 mL of acetic
CHARACTERS acid Rand 90 mL of distiUed water R.
Appearance
White or almost white, crystalline or granular powder. MagnesIum and alkali metals
Maximum 0.4 per cent (expressed as MgO).
Solubility
Dissolve 1.00 g in 100 mL of boiling wale/" R, add 10 mL of
Sparingly soluble in water, freely soluble in boilingwater.
ammonium chloride solutien R, 1 mL of ammonia Rand,
IDENTIFICATION dropwise, 50 mL of hot ammonium oxalate solution R. Allow
A. Thin-layer chromatography (2.2.27). to stand for 4 h, dilute to 200 mL with water R and filter.
Test solution Dissolve 20 mg of the substance to be Evaporate 100 mL of the filtrate to dryness and ignite.
examined in 1 mL of water R, heatingif necessary in a water- The residue weighs a maximwn of 2 mg.
bath at 60 "C. Loss on drying (2.2.32)
Reference solution Dissolve 20 mg of calcium gluconale CRS Maximum 2.0 per cent, determined on 1.000 g by drying in
in 1 mL of water R, heating if necessary in a water-bath at an oven at 105 "C for 16 h.
60 "C. Microbial contamination
Plate TLC silica gelplate R (5-40 JlIl1) [or TLC silica gel TAMC: acceptance criterion 10' CFU/g (2.6.1Z).
plate R (2-10 pm)]. TYMC: acceptance criterion 102 CFU/g (2.6.12).
Mobile phase concentrated ammonia R, ethyl acetate R,
ASSAY
wale/" R, ethanol (96 per cenl) R (10:10:30:50 VIVIVIV).
Dissolve 0.350 g in 20 mL of hot water R, allow to cool and
Application I pL. dilute to 300 mL with water R. Carry out the
Development Over 213 of the plate. complexometric titration of calcium (2.5.11).
Drying At 100 "C (qr 20 min, then allow to cool. 1 mL of 0.1 M 'odium edetate is equivalent to 43.04 mg
Detection Spray with a solution containing 25 gILof of C12H22CaOI4'
ammonium molybdate R and 10 gIL of cerium sulfate R in daute ~ PhE<r
,ulfuric acidR, and heat at 100-105 "C for about 10 min.
with the test solution is similar in position) colour and size to
reference solution.
B. Solution S (see Tests) gives the reactions of calcium
(2.3.1).
C. Loss on drying (see Tests).
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2022 Calcium Gluconate 1-399
pH (2.2.3)
Calcium Gluconate for Injection 6.4 to 8.3.
(ph. Bur. monograph 0979) Dissolve 1.0 gin 20 mL of carbon dioxide-free water R,
heating on a water-bath.
HO H co,: J Organic impurities and boric acid
~
. " --OH Introduce 0.5 g into a porcelain dish previously rinsed with
ea2+ HO' -, H H,O
[ HO H
sulfuric acid R and placed in a bath of iced water. Add 2 mL
HO 2 of cooled ru/fun'c acid R and mix, No yellow or brown colour
develops. Add I mL of chromotrope II B solution R. A violet
colour develops and does not become dark blue.
448.4 18016-24-5 The solution is not more intensely coloured than that of a
mixture of I mL of chromotrope II B solution Rand 2 mL of
Action and use
cooled sulfuric acid R.
Oxalates
Preparation
Calcium Gluconate Injection
PhE" _
examined in waterfor chromatography R and dilute to
DEFINITION 100.0 mL with the same solvent.
Calcium bis[(2R,3SAR,5R)-2,3A,5,6- Reference solution Dissolve 1.00 g of the substance to be
pentahydroxyhexanoate] monohydrate (calcium di examined in waterfor chromatography R, add 0.5 mL of a
(n-gluconate) monohydrate). 0-.152 gIL solution of sodium oxakueR in water for
chromaU>graphy R and dilute to 100.0 mL with the same
Content
solvent.
99.0 per cent to 101.0 per cent of CI2H22CaOl4>H20.
Precolumn:
CHARACfERS - size: 1= 30mm, 0;;;; 4 mmi
Appearance - stationary phase: suitable strong anion-exchange resin
White or almost white, crystalline or granular powder. (30-50 pm).
Solubility Columns 1 and 2:
Sparingly soluble in water, freely soluble in boiling water. - size: 1= 0.25 m, 0 = 4: nun;
IDENTIFICATION . - stationary phase: suitable strong anion-exchange resin
A. Thin-layer chromatography (2.2.27). (30-50 um).
Test solution Dissolve 20 mg of the substance to be Ardon-suppresser column Connected in series with the
examined in 1 mL of water R, heating if necessary in a water- precolumn and analytical columns and equipped with a
bath at 60 "C. micromembrane mat separates the mobile phase from the
suppressor regeneration solution, flowing countercurrent to
Reference solution Dissolve 20 mg of calcium glucona" CRS the mobile phase.
in 1 mL of water R, heating if necessary in a water-bath at
60 "C. Mobile phase Dissolve 0.212 g of anhydrous sodium
carbonate R and 63 mg of sodium hydrogen carbonate R in
PIa,. TLC silica gelpkue R (5-40 urn) [or TLC silica gel
waterfor chromau>graphy R and dilute to 1000.0 mL with the
plate R (2-10 ~m)J .. same solvent.
Mobile phase amcentrated ammonia R, ethylacetate R,
Flow m" of the mobile phase 2 mUmin.
waterR, ethanol (96 per eenO R (10:10:30:50 VIVIVIV).
Suppressor regeneration solution 1.23 gIL solution of sulfurU
Application I ~L. acidR in waterfor chromatography R.
Development Over 2/3 of the plate. Flow rale of the suppressor regeneration solution 4 mUmin.
Drying At 100°C for 20 min; allow to cool. Detection Conductance.
Detection Spray with a solution containing 10 gIL of cerium . Injution 50 ~L.
sulfate Rand 25 gIL of ammonium molybda,. R in dinue
sulfuric add R and heat at 105°C for about 10 min. System suitability Reference solution:
- repeatabjli~: maximum relative standard deviation of
Results After 5 min, the principal spot in the chromatogram 2.0 per cent for the area of the peak due to oxalate after 5
obtained with the test solution is similar in position, colour
injections.
with the reference solution. Inject 50 J1L of each solution 3 times. Calculate the content
of oxalates in parts per million using the following
B. About 20 mg gives reaction (b) of calcium (2.3.1). expression:
TESTS
Solution S
To 10.0 g add 90 mL of boiling distilled waterR and boil
with stirring, for not more than 10 5J until completely
ST area of the peak due to oxalate in the chromatogram obtained
dissolved, then dilute to 100.0 mL with the same solvent.
with the test solution;
Appearance of solution SR area of the peak due to oxalate in the chromatogram obtained
At 60 "C, solution S is not more intensely coloured than with the reference solution.
reference solution B 7 (2.2.2, Method II). After cooling to
20 "C, it is not more opalescent than reference suspension II Limit:
(2.2.1). - oxalates: maximum 100 ppm.
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1-400 Calcium Glycerophosphate 2022

Dissolve 0.5 g in a mixture of 2 mL of hydrochloric acidR1
Calcium Glycerophosphate
and 10 mL of water R. Boil for 5 min, allow to cool, add (Ph. Eur. monograph 0980)
10 mL of sodium carbonate solution R and allow to stand for
10 min. Dilute to 25 mL with water R and filter. To 5 mL of C,H,CaOoP 210.1
the filtrate add 2 mL of cupti-tanaric solution R and boil for
1 min. Allow to stand for 2 min. No red precipitate is Action and use
formed. Excipient.
Chlorides (2.4.4) P1>E<x _
Maximum 50 ppm.
DEFINITION
To 10 mL of previously filtered solution S add 5 mL of Mixrure in variable proportions of the calcium salt of (RS)-
water R. 2,3-dihydroxypropyl phosphate and of2-hydroxy-l-
Phosphates (2.4.11) (hydroxymethyl)ethyl phosphate which may be hydrated.
Maximum 100 ppm.
Content
Dilute 1 mL of solution S to 100 mL with water R. 18.6 per cent to 19.4 per cent of Ca (dried substance).
Sulfates (2.4.13)
CHARACTERS
Maximum 50 ppm, determined on previously filtered
Appearance
solution S.
Prepare the standard using a mixture of 7.5 mL of sulfate
Solubility
standard solution (10 ppm SO.! Rand 7.5 mL of distilled
water R. Sparingly soluble in water, practically insoluble in ethanol
(96 per cent).
Iron
Maximum 5 ppm. IDENTIFICATION
Atomic absorption spectrometry (2.2.23, Method l). A. Mix I g with I g of potasrium hydrogen sulfate R in a test
tube fined with a glass rube. Heat strongly and directthe
Test solution Introduce 2.0 g into a 100 mL
white vapour towards a piece of filter paper impregnated with
polyretraffuoroethylene beaker and add 5 mL of nitric acid R.
a freshly prepared 10 gIL solution of sodium nitroprusside R.
Boil, evaporating almost to dryness. Add I mL of S/TOUg
The filter paper develops a blue colour in contactwith
hydrogen peroxide solution R and evaporate again almost to
piperidine R.
dryness. Repeat the hydrogen peroxide treatment until a clear
solution is obtained. Using 2 mL of nimc acidR, transfer the B. Ignite 0.1 g in a crucible. Take up the residue with 5 mL
solution into a 25 mL volumetric flask. Dilute to 25.0 mL of nitric acid R and heat on a water-bath for 1 min. Filter.
with dilute hydrochloric add R. In the same manner, prepare a The filtrate gives reaction (b) of phosphates (2.3.1).
compensation solution using 0.65 g of calcium chloride R1 C. It gives reaction (b) of calcium (2.3.1).
instead of the substance to be examined. TESTS
Reference solutions Prepare the reference solutions from iron Solution S
standard solution (20 ppm Fe) R, diluting with diluUl Dissolve 1.5 g at room temperature in carbon dioxide-free
hydrochloric acid R. water R prepared from distilled water R and dilute 10 150 mL
Source Iron hollow-cathode lamp. with the same solvent.
Wavelength 248.3 om. Appearance of solution
Atomisation device Air-acetylene flame. Solution S is not more opalescent than reference
Carry out a basic correction using a deuterium lamp. suspension ill (2.2.1).
Magnesium and alkali metals Acidity or alkal1nlty

To 100 mL of solution S add 0.1 mL of phenolphthalein
solution R. Not more than 1.5 mL of 0.1 M hydrochloric acid
To 0.50 g add a mixture of 1.0 mL of dilute acetic acid Rand
or 0.5 mL of 0.1 M sodium hydroxide is required to change
10.0 mL of water R and rapidly boil, whilst shaking, until
the colourof the indicator.
completely dissolved. To the boiling solutionadd 5.0 mL of
ammonium oxalate solution R and allow to stand for at least Citric acid .
6 h. Filter through a sintered-glass filter (1.6) (2.1.2) into a Shake 5.0 g with 20 mL of carbon dioxide-free water Rand
porcelain crucible. Carefully evaporate the filtrate to dryness filter. To the filtrate add 0.15 mL of sulfuric acidR and filter
and ignite. The residue weighs not more than 2 mg. again. To the filtrate add 5 mL of mercuric sulfate solution R
and heat to boiling. Add 0.5 mL of a 3.2 gIL solution of
potassium pennanganate R and again heat to boiling.
Less than 167 IU/g.
No precipitate is formed.
Microbial contamination Glycerol and ethanol (96 per cent)-soluble substances
TAi\1C: acceptance criterion 10' CFU/g (2.6.12).
.Maximum 0.5 per cent.
ASSAY Shake 1.000 g with 25 mL of ethanol (96 per cen!! R for
Dissolve 0.350 g in 20 mL of hot water R, allow to cool and 1 min. Filter. Evaporate the filtrate on a water-bath and dry
dilute to 300 mL with water R. Carry out the the residue at 70°C for I h. The residue weighs a maximum
complexometric titration of calcium (2.5.11). Use 50 109 of of5 mg.
calamecavboxytic acid triturate R. Chlorides (2.4.4)
I mL of 0.1 .iH sodium edetate is equivalent to 44.84 mg Maximum 500 ppm.
of ClzH2ZCaOl4lHzO.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E<X
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2022 Calcium Hydrogen Phosphate 1-401
Dissolve 0.1 g in a mixture of 2 mL of acetic add Rand dissolve me precipitate and dilute to 50 mL with distilled
8 mL of water R and dilute to 15 mL with water R. water R.•
Phosphates (1.4.11) Acid-insoluble substances
Maximum 400 ppm. Maximum 0.2 percent.
Dilute 2.5 mL of solution S to 100 mL with water R. Dissolve 5.0 g in 40 mL of waterR, add 10 mL of
Sulfates (lA.H) hydrochlori< ocidR and heat to boiling for 5 min. Cool, then
Maximum 0.1 per cent, determined on solution S. collect the insoluble substances using ashless filter paper.
Wash with water R until turbidity is no longer produced
Arsenic u.cz, Me/hodA)
when silver nitrate solun"on R2 is added. Ignite me residue and
Maximum 3 ppm.
me filter paper at 600 ± 50 °C. The residue weighs not
Dissolve 0.33 g in water R and dilute to 25 mL with the more than 10 mg.
same solvent.
Carbonates
Iron (lA.9) Shake 1.0 g with 5 mL of corbon dioxide-free woterR and add
Maximum 50 ppm, detemined on 0.20 g. 2 mL of hydrochloric acidR. No effervescence is produced.
Loss on drying (1.1.31) Chlorides
Maximum 12.0 per cent, determined on 1.000 g by drying in Maximum 0.25 per cent.
Test solution Dissolve 0.20 g in a mixture of 20 mL of
ASSAY water Rand 13 mL of dilute nitricacid R by warming if
Dissolve 0.200 g in water R. Carry out the complexometric necessary, dilute to 100 mL with water R and filter if
titration of calcium (1.5. JI). necessary. Use 50 mL of this solution.
I mL of 0.1 M sodium edeuue is equivalent to 4.008 mg of Reference solution To 0.70 rnL of 0.01 M hydrochloric acid,
Ca. add 6 mL of dilu" nitric acidR and dilute to 50 mL with
____ ~ PhE" water R.
Add 1 mL of silvernitrate solution R2 to the test solution and
to the reference solution and mix. After standing for 5 min
protected from light, any opalescence in the test solution) by
Calcium Hydrogen Phosphate1 viewing vertically or horizontally against a black background,
is not more intense than that in the reference solution.
Anhydrous Calcium Hydrogen Phosphate tFluorides
(ph. Bur. monograph 0981) Maximum 100 ppm.
CaHPO, 136.1 7757-93-9 Potentiometry (1.1.36, Method II).
PhE" _ ehelau·ng solution Dissolve 45 g of cycJohexylenedinitriJoletra~
acetic acidR in 75 mL of sodium hydroxide so/u.ion Rand
DEFINlTION dilute to 250 mL with waterR.
Content Test sdution Dissolve 1.000 g in 4 mL of hydrochlori<
acidRl, add 20 mL of chelating solution, 2.7 mL of glacial
tCHARACTERS acetic acidRand 2.8 g of sodium chloride R, adjust to pH 5-6
Appearance with sodium hydroxide so/uwn R and dilute to 50.0 mL with
Whiteor almost white, crystalline powder, or colourless water R.
crystals. Reference soluwn Dissolve 4.42 g of sodiumj/uoride R,
SolublIlty previously dried at 300 °C for 12 h) in water R and dilute to
Practically insoluble in water and in ethanol (96 per cent). 1000.0 mL with the same solvent. Dilute 50.0 mL of this
It dissolves in dilutehydrochloric acid and in dilute nitric solution to 500.0 mL with unal-ionic-strength-odjustment
acid.• buffer R (200 ppm F).
IDENTIFICATION Indicator electrode Fluoride-selective.
A. Dissolve with heating 0.1 gin 10 mL of dilute hydrochloric Reference dec/rode Silver-silver chloride.
acidR. Add 2.5 mL of dilute ommonia Rl, shake, and add Carry out the measurement on 20.0 mL of the test solution.
5 mL of a 35 gIL solution of ammonium oxalate R. A white Add at least 3 times 0.10 mL of the reference solution and
precipitate is produced. carry out the measurement after each addition. Calculate the
B. Dissolve 0.1 gin 5 mL of dihue nitric acid R, add 2 mL of concentration of fluorides using the calibration cwve.•
ammonium molybdau solution R andheat at 70 °C for Sulfates
1-2 min. A yellowprecipitate is produced. Maximum 0.5 percent.
OC. It complies with me limits of me assay.O Test So/UIUm Dissolve 0.5 g in a mixture of 5 mL of water R
TESTS and 5 mL of dilu,. hydrochloric ocidR and dilute to 100 mL
with waler R. Filter if necessary. To 20 mL of this solution,
tSolution S
add I mL of dilu" hydrochloric acidR and dilute to 50 mL
Dissolve 2.5 g in 20 mL of dilute hydrochloric acidR, filter if
necessary and add dilute ammonia Rl untila precipitate is with water R.
formed. Add just sufficient diiute hydrochloric acid R to Reference solsuion To 1.0 rnL of 0.005 M su/furic acid, add
I rnL of dilu" hydrochloric ocidR and dilute to 50 mL with
water R. Filter if necessary.
To the test solution and to the reference solution, add 2 mL
I This monograph has undergone the pnxess of Pharmacopoeial
of a 120 gIL solution of barium chloride R and allow to stand
harmonisation. SeechaPter 5.8. Pharm(U(}jJOeial harmonisation.
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1-402 Calcium Hydrogen Phosphate Dihydrate 2022
for 10 min. Any opalescence in the test solution, by viewing

Calcium Hydrogen Phosphate ***
vertically or horizontally against a black background) is not *** ***
more intense thanthat in the reference solution. Dihydrate ***
tArsenic (2.4.2, Method A) Dibasic Calcium Phosphate
Maximum 10 ppm, determined on 2 mL of solution S.• (Ph. Eur. monograph 0116)
Barium NOTE: The nameCalcium Hydrogen Phosphate wasfonntrly used
To 0.5 g, add 10 mL of wal<r R and heat to boiling. While in the UnitedKingdom.
stirring, add I mL of hydlYXhloric acidR dropwise. Allow to
CaHPO..,2H,O 172.1 7789-77-7
cool and filter if necessary. Add 2 mL of a 10 gIL solutionof
PhE" _
dipotassium sulfate R and allow to stand for 10 min.
No turbidity is produced. DEFINITION
tIron (2.4.9) Content
Maximum 400 ppm. 98.0 per cent to 105.0 per cent.
Dilute 0.5 mL of solution S £0 10 mL with water R.• CHARACTERS
Loss on ignition Appearance
6.6 per cent to 8.7 per cent, determined on 1.000 g to Wh,ite or almost white, crystalline powder.
constant mass at 800-825 "C. Solubility
ASSAY Practically insoluble in water and in ethanol (96 per cent).
Dissolve 0.4 g in 12 mL of dilul< hydrochloric acid R by It dissolves in dilute hydrochloric acid and in dilute nitric
heating on a water bath if necessary and dilute (Q 200.0 mL acid.
with wat<r R. To 20.0 mL of this solution add 25.0 mL of IDENTIFICATION
0.02 M sodium edeuue, 50 mL of water R, 5 mL of ammonium A. Dissolve with heating 0.1 gin 10 mL of dilute hydrochloric
chloride buffer solution pH 10.7 R and about 25 mg of mordant acidR. Add 2.5 mL of dilute ammonia R1, shake and add
black 11 tn"curate R. Titrate the excess of sodium edetate with 5 mL of a 35 gIL solution of ammonium oxalate R. A white
0.02 M zinc sulfare. Carry out a blank titration. precipitate is produced.
1 mL of 0.02 M sodium edetate is equivalent to 2.721 mg B. Dissolve 0.1 gin 5 mL of dilul< miricacidR, add 2 mL of
ofCaHPO•. ammonium molybdate solution R and heat at 70°C for 2 min.
FUNCTIONAlJTY-RELATED CHARACTERISTICS A yellowprecipitate is produced.
This section prouides information on charactenstia that an' C. It complies with the limits of the assay.
recognised as being relevant ccntrol parameters for oneor more TESTS
functions of the substance when used as an excipient (see chapter
Solution S
Dissolve 2.5 g in 20 mL of dilute hydrochloric acidR, filter if
related charactetistia section may alsobepresent in the mandatory
necessary and add dilute ammonia Rl until a precipitate is
part 0/the monograph since they alsorepresent mandatory quali~
formed. Add just sufficient d,lul< hydrochlon'c acid R to
criteria: In such cases, a cross-reference to the rests described in the
dissolve the precipitate and dilute to 50 mL with distilled
mandatory part is indudedin theFunctionality-related
water R.
characteristics section. Control of the characteristics can contribut«
'" the quoli"" of a medicinal product by improving the consistency Acid-insoluble substances
of the mantifacturing process and the perfonnanee of the medicinal Maximum 0.2 per cent.
product during use. Where comrd methods are cited, they are Dissolve 5.0 g in 40 mL of wat<r R, add 10 mL of
recognised as being suitable for thepurpose, but orher methods can hydlYXhloric acid R and heat to boiling for 5·min. Cool, then
also be used. Wherever remIts for a particular characteristic are collect the insoluble substances using ashless filter paper.
reported, the control method must be indicated: Washwith water R until turbidity is no longer produced
Thefollowing characteristics may be relevant for calcium hydrogen when Silver nitrate solution R2 is added to the filtrate. Ignite at
phosphal< usedasfiller in tablets and capsules. 600 ± 50 "C. The residue weighs not more than 10 mg.
Particle-size distribution (2.9.31 or 2.9.38) Carbonates
Shake 0.5 g with 5 mL of carbon dioxide-free wat<r R and add
Bulk and tapped density (2.9.3<{)
1 mL of hydrochloric acidR. No effervescence is produced.
Powder flow (2.9.36)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" Chlorides
Test solution Dissolve 0.20 g in a mixture of 20 mL of
wat<r Rand 13 mL of dilul< nitric acidR by warming if
necessary, dilute to 100 mL with water R and filter if
necessary. Use 50 mL of this solution.
Reference solution To 0.70 mL of 0.01 M hydrochloric acid,
add 6 mL of dilute nitric acid R and dilute to 50 mL with
water R.
Add 1 mL of silver nitrate solution R2 to the test solution and
to the reference solution and mix. After standing for 5 min
protected from light, any opalescence in the testsolution is
not more intense than that in the reference solution.
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2022 Calcium Hydroxide 1-403
Fluorides FUNCTIONALITY-RELATED CHARACTERISTICS

Maximum 100 ppm. This section provides infonnalion on characteristics that are
Potentiometry (2.2.36, MethodIf). recognised as being relevant control parameters for oneor more
Chelaring solution Dissolve 45 g of cyclohexylenedinitrilotelra- junctions of the substance when used as an excipient (see chapter
acetic acid R in 75 mL of sodium hydroxide solution Rand 5.15). Someof the charaaetistia described it, the Fimaionoluy-
dilute (0 250 mL with water R.
related characteristics seaion mc::ry alsobepresent in the mandatory
part of the monograph since they also represent mandatory quaNty
Test solution Dissolve 1.000 g in 4 mL of hydrochloric cntena. In such cases) a cross-reference to the tests described ill the
acid R1, add 20 mL of chelating solution, 2.7 mL of glacial mandatory part is included in the Funaionality-rekued
acetic acidRand 2.8 g of sodium thloride R, adjust to pH 5-6 characteristics section. Control of the characteristics can contribute
with sodium hydroxide sdsuion R and dilute to 50.0 rnL with to the qualityof a medicinal product by improving the consistency
water R. of the monvfaaunng process and the peformance of the medicinal
Reference sonaion Dissolve 4.42 g of sodium fluoride R, product during use. W'here «mtrolmethods are cited) they are
previously dried at 300 DC for 12 h, in water R and dilute to recognised as being suitable for thepurpose) but other methods can
1000.0 mL with the same solvent. Dilute 50.0 rnL of this alsobe used. W'herever results for a particular characteristic are
solution to 500.0 mL with UJuzl-ionic-slYtngth-adjustment reported) the control method must be indicated.
buffer R (200 ppm F). The following characteristics may be relevant for calcium hydrogen
Indicator electrode Fluoride-selective. phosphate dihydrate usedas filler in tablets and capsules.
Reference electrode Silver-silver chloride. Particle-size distribution (2.9.31 or 2.9.38)
Carry out the measurement on 20.0 mL of the test solution. Bulk and lapped density (2.9.34)
Add at least 3 times 0.10 mL of the reference solution and
carry out the measurement after each addition. Calculate the Powder flow (2.9.36)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1M"
concentration of fluorides using the calibration curve.
Sulfates
Test solution Dissolve 0.5 g in a mixture of 5 mL of waterR Calcium Hydroxide ***
and 5 mL of dilute hydrochloric acid Rand dllute 10 100 rnL *** ***
with water R. Filter if necessary. To 20 mL of this solution, (ph. Eur. monograph 1078) ***
add 1 mL of dilute hydrothlori< acidR and dilute to 50 mL
with water R. Ca(OH), 74.1 1305-62-0
Reference soluewn To 1.0 rnL of 0.005 M sulfuric acid, add Preparation
1 rnL of dilute hydroch/Qric acidR and dilute to 50 rnL with Calcium Hydtoxide Solution
water R. Filter if necessary. POE" _
To the test solution and to the reference solution, add 2 mL DEFINITION

of a 120 gIL solution of barium chloride R and allow to stand Content
for 10 min. Any opalescence in me test solution is not more 95.0 per cent to 100.5 per cent.
intense than that in the reference solution.
CHARACTERS
Arsenic (2.4.2, MethodA)
Appearance
lYlaximum 10 ppm, determined on 2 mL of solution S.
Whiteor ahnost white, fine powder.
Barium
SolublUty
To 0.5 g, add 10 mL of waterR and heal to boiling. While
Practically insoluble in water.
stirring, add 1 rnL of hydrochlori< acidR dtopwise. Allow to
cool and filter if necessary. Add 2 rnL of a 10 gIL solution of IDENTIFICATION
dipotassium sulfate R and allow to stand for 10 min. A. To 0.80 g in a mortar, add 10 mL of water Rand 0.5 mL
No tusbidity is produced. of phenolphthalein solution R and mix. The suspension turns
Iron (2.4.9) red. On addition of 17.5 rnL of 1 M hydrochlori< acid, the
Maximum 400 ppm. suspension becomes colourless without effervescing. The red
colour occursagain when the mixture is trirurated for 1 min.
Dilute 0.5 mL of solution S to 10 mL with waterR.
On addition of a further 6 mL of 1 M hydrochloric acid and
Loss on ignition triturating, the solution becomes colourless.
24.5 per cent to 26.5 per cent, determined on 1.000 g by B. Dissolve ahout 0.1 g in dilute hydrochlori< acidR and dilute
ignition to constant mass at 800-825 °C. to 10 mL with water R. 5 mL of the solutiongive
ASSAY reaction (h) of calcium (2.3.1).
Dissolve 0.4 g in 12 rnL of dilute hydrochloric acidR by TESTS
heating on a water bath if necessary and dilute to 200 mL Matter insoluble in hydrochloric acid
with waterR. To 20.0 rnL of this solution add 25.0 rnL of Maximum 0.5 per cent.
0.02 iW sodium edetate, 50 mL of water RJ 5 mL of ammonium
chloride buffer sduuon pH 10.7 R and about 25 mg of mordant Dissolve 2.0 g in 30 mL of hydrochloric acidR. Boil the
bkuk 11 tritunue R. Titrate the excess of sodium edetate with solution and filter. Washthe residue with hot water R.
0.02 M zinc sulfate. Carry out a blank titration. The residue weighs a maximum of 10 mg.
1 mL of 0.02 M sodium edetate is equivalent to 3.44 mg Carbonates
of CaHPO.,,2H2 0 . Maximwn 5.0 per cent of CaC03 •
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1-404 Calcium Lactate 2022
Add 5.0 mL of 1 M hydrochloric acid to the titrated solution Solublllty

obtainedunder Assay and titrate with 1 lU sodium hydroxide Soluble in water, freely soluble in boiling water, very slightly
using 0.5 mL of methyl orange solution R as indicator. soluble in ethanol (96 per cent).
I mL of 1 M hydrochloric acid is equivalent to 50.05 mg IDENTIFICATION
ercsco, A. Loss on dsying (see Tests).
Chlorides (2.4.4) B. It gives the reaction of lactates (2.3.1).
Maximum 330 ppm. C. It gives reaction (b) of calcium (2.3.1).
Dissolve 0.30 g in a mixture of 2 mL of nimc acid Rand
TESTS
10 mL of water R and dilute to 30 mL with foarer R.
Solution S
Sulfates (2.4.13) Dissolve s.o g with heatingin carbon dioxide-free water R
Maximum 0.4 per cent. prepared from distiOed water R, allow to cool and dilute to
Dissolve 0.15 g in a mixture of 5 rnL of dilure hydrochloric 100 mL with the same solvent.
acidR and 10 mL of distilled water R and dilute to 60 rnL Appearance of solution
with distilled warer R. Solution S is not more opalescent than reference
Arsenic (2.4.2, Methad A) suspension II (2.2.1) and not more intensely coloured than
Maximum 4 ppm. reference solution BY6 (2.2.2, MethodII).
Dissolve 0.50 g in 5 rnL of brominared hydrochloric acid Rand Acidity or alkalinity
dilute to 50 rnL with water R. Use 25 rnL of this solution. To 10 rnL of solution S add 0.1 mL of phenolphthalein
Magnesiwn and alkall metals solution R and 0.5 rnL of 0.01 M hydrochloric acid.
Maximum 4.0 per cent calculated as sulfates. The solution is colourless. Not more than 2.0 mL of 0.01 M
Dissolve 1.0 g in a mixture of 10 rnL of hydrochloric acid R sodium hydroxide is required to change the colour of the
and 40 mL of warer R. Boil and add 50 rnL of a 63 gIL indicator to pink.
solution of oxalic acidR. Neutralise with ammonia Rand Chlorides (2.4.4)
dilute to 200 mL with water R. AUow to stand for I hand Maximum 200 ppm.
filter through a suitable filter. To 100 mL of the filtrate, add Dilute 5 rnL of solution S to 15 rnL with worer R.
0.5 mL of sulfuric acid R. Cautiously evaporate to dryness
Sulfates (2.4.13)
and ignite. The residue weighs a maximum of 20 mg.
Maximum 400 ppm.
ASSAY Dilute 7.5 mL of solution S to 15 mL with distiUed worer R.
To 1.500 g in a mortar, add 20-30 mL of water Rand
Barium
0.5 mL of phenolphthalein solution R. Titrate with 1 M
To 10 rnL of solution S add I rnL of calcium sulfate
hydrochloric acid by triturating the substance until the red
solution R. Allow to stand for IS min. Any opalescence in the
colour disappears. The final solution is used in the tests for
solution is not more intense than that in a mixture of I mL
carbonates.
of dis'i1kd water Rand 10 rnL of solution S.
I mL of 1 M hydrochlori< acid is equivalent to 37.05 mg
ofCa(OH),. Iron (2.4.9)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE«
Maximum 50 ppm.
Dilute 4 rnL of solution S to to mL with woter R.
Magnesium. and alkali salts
Maximum I per cent.
Calcium Lactate To 20 rnL of solution S add 20 mL of water R, 2 g of
ammonium chloride Rand 2 mL of dl1ute ammonia RI. Heat to
Anhydrous Calciwn Lactate boiling and rapidly add 40 mL of hot ammonium oxalate
(Ph. Bur. monograph 2118) sdtuion R. Allow to stand for 4 h, dilute to 100.0 mL with
water R and filler. To 50.0 rnL of the filtrate add 0.5 mL of
su/furk acid R. Evaporate to dryness and ignite the residue to
and enanliomer constant mass at 600 ± SO °C. The residue weighs a
maximum of S'mg.
218.2 Maximum 3.0 per cent, determined on 0.500 g by drying in
an oven at 12S "C.
Action and use
Used in treatment of calcium deficiency. ASSAY
Dissolve 0.200 g in water R and dilute to 300 mL with the
PIIE« _
same solvent. Carry out the complexometric titration of
DEFINITION calcium (2.5.11).
Calcium bis(2-hydrnxypropanoate) or mixture of calcium I mL of 0.1 M sodium edeuue is equivalent to 21.82 mg
(2R)-, (2S)- and (2RS)-2-hydrnxypropanoates. of C.HloCaO•.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE«
Content
CHARACTERS
Appearance
White or almost white, crystalline or granular powder.
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2022 Calcium Lactate 1-405
sulfun'c acid R. Evaporate to dryness and ignite the residue to

Calcium Lactate Monohydrate constant mass at 600 ± 50 °C. The residue weighs a
(Ph. Eur. monograph 2117) maximum of 5 mg.
Ca,. [H3Cy~-j andenantiomer . ~o 5.0 per cent to 8.0 per cent, determinedon 0.500 g by
drying in an oven at 125 °C.
H OH 2
ASSAY
C,HIOCaO"H,O 236.2 Dissolve a quantity equivalentto 0.200 g of the dried
substance in waler R and dilute to 300 mL with the same
Action and use solvent. Carry out the complexomettic titration of calcium
Used in treatment of calcium deficiency. (2.5.11).
PhE" _ 1 mL of 0.1 M sodium edetate is equivalent to 21.82 mg
of C,HIOCaO,.
DEFINITION _ _ _ _ _ _ _ _ _ _ _ _ _~ PI>E"
Calcium bis[(2E)-2-hydroxypropanoate] or mixrure of
calcium (2R)-, (2S)- and (2RS)-2-hydroxypropanoates
monohydrates.
Content Calcium Lactate Pentahydrate
CHARACTERS Calcium Lactate
Appearance (Ph. Eur. monograph 0468)
White or almost white, crystalline or granular powder.
Solubility Ca" [H'C'F..v'"', co,-j and enantiomer • 5 H:!O
Soluble in water, freely soluble in boiling water,very slightly H OH 2
IDENTIFICATION CoHIOCaO,,5H,O 308.3
A. Loss on drying (see Tests).
Action and use
B. Ir gives the reaction of lactates (2.3.1).
C. It gives reaction (b) 'of calcium (2.3.1).
Preparations
TESTS Calcium and ErgocalciferolTablets
Solution S Calcium Lactate Tablets
Dissolve 5.4 g (equivalent to 5.0 g of the dried substance)
with heating in carbon dioxide-free water R prepared from Calcium and Ergocalciferol Chewable Tablets
distif!ed waterR, allow to cool and dilute to 100 mL with the PhE" -'- _
same solvent.
DEFINITION
Appearance of solution Calcium bis[(2E)-2-hydroxypropanoate) or mixrure of
Solution S is not more opalescent than reference calcium (2R)-, (2S)- and (2RS)-2-hydroxypropanoates
suspension n (2.2.1) and not more intensely coloured than pentahydrates,
reference solution BY, (2.2.2, Method If).
Content
AcIdity or alkalinity 98.0 pet cent to 102.0 per cent (driedsubstance).
To 10 mL of solution S add 0.1 mL of phenolphthalein
solution Rand 0.5 mL of 0.01 M hydrochloric acid. CHARACTERS
The solution is colourless. Not more than 2.0 mL of 0.01 M Appearance
sodium hydroxide is requiredto change the colour of the White or almost white, crystalline or granular powder,
indicator to pink. slightly efflorescent.
Chlorides (2.4.4) Solublllty
Maximum 200 ppm. Soluble in water, freely soluble in boiling water, very slightly
Dilute 5 mL of solution S to 15 mL with water R. soluble in ethanol (96 per cent).
Sulfates (2.4.13) IDENTIFICATION
Maximum 400 ppm. A. Loss on drying (see Tests).
Dilute 7.5 mL of solution S to 15 mL with distilled water R. B. It gives the reaction of lactates (2.3.1).
Iron (2.4.9) C. It gives reaction (b) of calcium (2.3.1).
Maximum 50 ppm. TESTS
Dilute 4 mL of solution S to 10 mL with water R. Solution S
Magnesium and alkali salts Dissolve 7.1 g (equivalent to 5.0 g of the dried substance)
Maximum 1 per cent. with heating in carbon dioxide-free waterR prepared from
To 20 mL of solution S add 20 mL of water R, 2 g of
distilled water R, allow to cool and dilute to 100 mL with the
same solvent.
ammonium chloride Rand 2 mL of dilute ammonia R1. Heat to
boiling and rapidly add 40 mL of hot ammonium oxalate
solution R. Allow to stand for 4 h, dilute to 100.0 mL with
water R and filter. To 50.0 mL of the filtrate add 0.5 mL of
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1-406 Calcium Lactate 2022
Appearance of solution Content

Solution S is not more opalescent than reference 98.0 per cent to 102.0 per cent (dried substance).
suspension II (2.2.1) and not more intensely colouredthan CHARACTERS
reference solution BY6 (2.2.2.1 lWemod II).
Appearance
Acidity or alkallnlty White or almost white, crystalline or granular powder.
To 10 rnL of solution S add 0.1 rnL ofphenolphlhalein
Solubility
solulion Rand 0.5 rnL of O. 01 M hydrochloric acid.
Soluble in water, freely soluble in boiling water, veryslightly
The solution is colourless. Not more than 2.0 mL of 0.01 M
soluble in ethanol (96 per cent),
sodium hydroxide is required to change the colour of the
indicator to pink. IDENTIFICATION
Chlorides (2.4.4) A. Loss on drying (see Tests).
Maximum 200 ppm. B. It gives the reaction of lactates (2.3.1).
Dilute 5 mL of solution S to 15 mL with water R. C. It gives reaction (b) of calcium (2.3.1).
Sulfates (2.4.13) TESTS
Maximum 400 ppm. Solution S
Dilute 7.5 rnL of solution S '0 15 rnL with distilled waler R. Dissolve 6.2 g (equivalent '0 5.0 g of the dried substance)
with heating in carbon dioxide-free water R prepared from
Iron (2.4.9)
distined water R, aUow to cool and dilute to 100 mL with the
Maximum 50 ppm.
same solvent.
Magnesium and aIkall salts Solution S is not more opalescent than reference
Maximum 1 per cent. suspension II (2.2. J) and not more intensely coloured than
To 20 mL of solution S add 20 mL of waler R, 2 g of reference solution BY. (2.2.2, Melhod 11).
ammonium chloride Rand 2 mL of dilute ammonia RI. Heat to Acidity or alkallnlty
boiling and rapidly add 40 rnL of hot ammonium oxalal~ To 10 mL of solution S add 0.1 mL of pherwlphlha/ein
solution R. Allow to stand for 4 h, dilute to 100.0 mL with sdution R and 0.5 mL of O. 01 M hydrochloric acid.
waW" Rand filrer. To 50.0 rnL of the filtrate add 0.5 mL of
The solution is colourless. Not more than 2.0 mL of 0.01 M
sulfuric acid R. Evaporate to dryness and ignite the residue to sodium hydroxide is required to change the colour of the
constant mass at 600 ± 50 °C. The residue weighs a
indicator to pink.
maximum of 5 mg.
Chlorides (2.4.4)
Maximum 200 ppm.
drying in an oven at 125 °C. Dilute 5 mL of solution S to 15 mL with Water R.
Sulfates (2.4.13)
ASSAY
Maximum 400 ppm.
Dissolve a quantity equivalent to 0.200 g of the dried
substance in water R and dilute to 300 mL with the same Dilute 7.5 rnL of solution S to 15 mL with dislil/ed waler R.
solvent. Carry out the complexomerric titration of calcium Iron (2.4.9)
(2.5.11). Maximum 50 ppm.
'0
I rnL of 0.1 M sodium edeuue is equivalent 21.82 mg Dilute 4 mL of solution S to 10 mL with water R.
of C.,HlOCaO•. Magnesium and alkali salts
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" Maximum 1 per cent.
To 20 mL of solution S add 20 rnL of waler R, 2 g of
ammonium chloride R and 2 mL of dllule ammonia R1. Heat to
boilingand rapidly add 40 mL of hot ammonium oxalate
Calcium Lactate Trihydrate solution R. AUow to stand for 4 h, dilute to 100.0 mL with
waler R and filter. To 50.0 mL of the filtrate add 0.5 rnL of
(Ph. Eur. monograph 0469) sulfuric acid R., Evaporate to dryness and ignite the residue to
constant mass at 600 ± 50 "C. The residue weighs a
maximum of 5 mg.
drying in an oven at 125 °C.
CoHIOCaO.,3H,O 272.3
ASSAY
Action and use Dissolve a quantity equivalent to 0.200 g of the dried
Used in treatment of calcium deficiency. substance in water R and dilute to 300 mL with the same
Preparation solvent. Carry out the complexometric titration of calcium
Calcium Lactate Tablets (2.5.11).
1 mL of O. J 1\1" sodium edetare is equivalent to 21.82 mg
PhE" _
of C6Hl0Ca06'
DEFINITION _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Calcium bis[(2B)-2-hydroxypropanoa,e] or mixture of
calcium (2R)-, (2S)- and (2RS)-2-hydroxypropanoates
trihydrates.
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2022 Calcium Levofolinare 1-407
Calcium Levofolinate Hydrate ******

*<,»
acid monohydrate R previously adjusted to pH 8 with
ammonia R,
Calcium Levofolinate Pentahydrate Application 5 ~L.
(Ph. Eur. monograph 1606) Development Over 2/3 of the plate.
Drying In air.
reference solution.
D. It gives reaction (b) of calcium (2.3.1).
C,oH2I CaN,O,,xH,O 511.5 TESTS

(anhydrous substance) Cany out the tests and the assayas rapidly as possible, proteaed
from actini« light.
Action and use Solution S
Antidote to folic acid antagonists. Dissolve 0.40 g in carbon dioxide-free waterR, heating at
PIlE" ~~ _ 40°C if necessary, and dilute (0 50.0 mL with the same
solvent.
DEFINITION Appearance of solution
Calcium (2S)-2-[4-[[[(6S)-2-amino-5-formyl-4-oxo- Solution S is clear (2.2.1).
1,4,5,6,7,8-hexabydropteridin-6-yl)methyl]amino]benzamido]
pH (2.2.3)
pentanedioatehydrate.
Content
- calcium leuofolinate (CwH21CaN,O,; M, 511.5):
-15.0 to -10.0 (anhydrous substance), measured at 25 "C.
97.0 per cent to 102.0 per cent (anhydrous substance);
- caldum (Ca; A c 40.08): 7.54 per cent (Q 8.14 per cent Dissolve 0.200 g in tris(hydroxymethyQaminomethane
(anhydcous substance). solution R previously adjusted to pH 8.1 with sodium hydroxide
solution R or hydrochloric acid Rl and dilute to 20.0 mL with
the same solvent.
CHARACTERS
Absorbance (2.2.25)
Appearance Maximum 0.25, determined at 420 nm on solution S.
White or light yellow, amorphous or crystalline, hygroscopic
powder. Ethanol
Head-space gas chromatography (2.2.28): use the standard
Solublllty
additions method.
Slightly soluble in water, practically insoluble in acetone and
in ethanol (96 per cent).
Test solutWn Dissolve 0.25 g of the substance to be
examined in waler R and dilute to 10.0 mL with the same
solvent.
IDENTIFICATION Reference solution Dilute 0.750 g of anhydrous ethanol R to
First identification: A, B, D. 1000.0 mL with water R.
Second identification: A, C, D. Column:
A. Specific optical rotation (see Tests). - material: fused silica;
B. Infrared absorption spectrophotometry (2.2.24). - size: I;;;; 10 m, 0 ;;;; 0.32 mm;
- stationary phase: sryrene-divinylbenzene copolymer R.
Comparison: calcium folinare CRS.
Cartier gas nitrogenfor chromatography R.
substance to be examined and the reference substance Flow rare 4 mUmin.
separately in the minimum quantity of water R and add Static head-space conditions thai may be used:
dropwise sufficient aarone R to produce a precipitate. Allow - equilibration temperature: 80 DC;
to stand for IS min, collect the precipitate by centrifugation, - equilibration time: 20 min;
wash the precipitate twice with a minimum quantity of - pressurisation time: 30 s.
acerone R and dry. Record new spectra using the residues. Temperature:
Test solution Dissolve 15 mg of the substance to be Time Temperature
examined in a 3 per cent VIV solution of ammonia Rand
(mln) CC)
Column 80 -> 220
Injection port 110
Reference solution Dissolve 15 mg of calcium folinate CRS in
Detector 270
a 3 per cent VIV solution of ammonia R and dilute to 5 mL
Plate cellulose for chromatography F254 R as the coating
substance. Injection At least 3 times.
Mobik phase The lower layer of a mixture of I volume of Limit:
isoamyl akohol Rand 10 volumes of a 50 gIL solution of curie - ethanol: maximum 3.0 per cent.
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1-408 Calcium Levofolinate 2022
Related substances Test solution Dissolve 50.0 mg of the substance to be

Liquid chromatography (2.2.29). examined in water R and dilute to 100.0 mL with the same
Prepare the solutions immediately before use. solvent.
Testsolution Dissolve 10.0 mg of the substance to be Reference solution (aJ Dissolve 10.0 mg of calcium
examined in water R and dilute to 10.0 mL with the same folinate CRS in water R and dilute to 20.0 mL with the same
solvent. solvent.
Reference solution (a) Dissolve 10.0 mg of calcium Reference solution (1)) Dilute 1.0 mL of reference solution (a)
folinate CRS in water R and dilute to 10.0 mL with the same to 100.0 mL with water R.
solvent. Column:
Reference solution (1)) Dilute 1.0 mL of the test solution to - size: 1= 0.15 m, (2) = 4 mm;
100.0 mL with u'Our R. Dilute 1.0 mL of this solution to - stationary phase: human albumin coated silicagel for dural
10.0 mL with waterR. separation R (5 Ilm);
Reference solution (c) Dissolve 5 rugof calcium jolinate for
system suitahili!Y CRS (containing impurities A, E J F and I) in Mobile phase Dissolve 9.72 g of sodium dihydrogen
5 mL of water R. phosphate R in 890 mL of waterfor chromatography Rand
adjust to pH 5.0 with sodium hydroxide solution R; add
Column:
100 mL of 2-propanol Rand 10 mL of acetonitrile R.
- size: 1 = 0.25 ill, 0 = 4 mm;
- srationary phase: end-capped oetadecylsi/yl silica gelfor Flow rate 1 mUmin.
chromatography R (5 ~m); Detection Spectrophotometer at 286 run.
- temperature: 40°C. Injection 10 ~L
Mobile phase Mix 165 mL of methanolRand 835 mL of a Retention times Levofolinic acid = about 9 min;
solutioncontaining 4.0 mL of tetrabutylammonium dihydrogen impurity H =about 19 min.
phosphate solution Rand 1.42 g of dis<>dium hydrogen phosphate System suiwbility:
dihydrate R in waterfor chromatography R, previously adjusted - resolution: minimum 5.0 between the peaksdue to
to pH 7.7 with phosphoric acid R or dilute sodium hydroxide levofolinic acid and impurity H in the chromatogram
solution R. obtained with reference solution (a). The sum of the areas
Flow rate 1.25 mllrnin. of the 2 peaks is 100 per cent. The peak area of
Detection Spectrophotometer at 254 nm. impurity H is 48 per cent to 52 percent. In the
Injutiun 10 ilL of the test solution and reference chromatogram obtainedwith reference solution (b)
solutions (b) and (c). . 2 clearly visible peaks are obtained.
Run time 4 times the retention time of folinic add. Limiu
Identification of impurities Use the chromatogram supplied - impurity H: maximum 0.5 per cent.
with calcium folinate for system suirabJ7ity CRS and the Chlorides
chromatogram obtained with reference solution (c) to identify Maximum 0.5 per cent.
the peaksdue to impurities A, E, F and I. Dissolve 0.300 g in 50 mL of waterR heating at 40 "C if
Relative retention With reference to folinic acid (retention necessary. Add 10 mL of dilute nitric acid R and titrate with
time =about 13.9 min): impurity E =about 0.4; 0.005 M silvernitrate determining the end-point
impurity A = about 0.6; impurity F = about 0.7; potentiometrically (2.2.20).
impurity 1 = about 1.2. l mL of 0.005 1\1 silvernitrate is equivalent to 0.177 mg of
System suitabiliry Reference solution (c): CI.
- resolution: minimum 2.0 between the peaks due to Water
impurities A and F. (2.5.12): 10.0 per cent to 17.0 per cent.
Calculation of percentage contents: Dissolve 0.100 g in a mixture of 15 mL offormamide R and
- correction factors: multiply the peak areas of the foUowing 50 mL of the titration solvent. Stirfor about 6 min before
impurities by the corresponding correction factor: titrating and use a suitable titrant that does not contain
impurity A = 0.6; impurity E = 0.6; impurity F = 0.6; pyridine.
=
impurity 1 0.6;
ASSAY
- for each impurity) use the concentration of calcium
levofollnate hydrate in reference solution (b). Cany oul the assays as rapidly as possible) prorected from actinic
lighc
Limits:
- impUJ;ties A) E: for each impurity) maximum 0.3 per cent; Calcium
- ;mpun't.ies F) I: for each impurity) maximum 0.2 per cent; Dissolve 0.400 g in 150 mL of waterR and dilute to 300 mL
- unspedfied impurities: for each impurity, maximum with the same solvent. Carry out the complexometric
0.20 per cent; titration of calcium (2.5.11).
- total: maximum 1.5 per cent; I mL of 0.1 M sodium ederate is equivalent to 4.008 mg of
- reponing threshold: 0.05 per cent. Ca.
The thresholds indicated under Related substances Calcium follnate
(Table 2034.-1) in the general monograph Substances for Liquid chromatography (2.2.29) as described in the test for
pharmaceutical use (2034) do not apply. related substances with the following modification.
Impurity H Injection Test solution and reference solution (a).
Liquid chromatography (2.2.29): use the normalisation Calculate the percentage content of C2oH21CaN707 taking
procedure. into account the assigned content of calcium foUnale CRS.
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2022 Calcium Levulinate Dihydrate 1-409
STORAGE o H C02.H
In an airtight container, protected from light. If the substance o ~~~C02H

is sterile, the container is also sterile and tamper-evident.
NYv-N'Y""N JJ
LABELLING
The label states, where applicable, that the substance is ~
.ii..
~ ~
tHO

preparations. F. (2S)-2- [4-[[(2-amino-4-oxo-1,4,7.8-tetrahydroptetidin-6-
IMPURITIES yl)methyl](fonnyl)amino]benzamido)pentanedioic acid
Specified impuniies A, E, F, H, I. (lO-fonnyldihydrofolic acid),
cntetion for other/unspecified impurities. It is therefore not
n«essary to idenufy these impurilies for demonstration of
compliance. See also 5.10. Control of itnpun"ties in substances for
pharmaceutical use) B, CJ D, G.
o H COzH G. (2S)-2- [4-[[(2-amino-4-oxo-I,4,7.8-tetrabydropteridin-6-
~~~CO,H yl)methyl]amino]benzamido]pentanedioic acid

(dihydrofolic acid).
H,NJJ
A. (2S)-2-(4-aminobenzamido)pentanedioic acid,
o H ,CO,H
o 9HO ~~~CO,H
NYv-N~NJJ
)l__ ~ ) <, H tHO H. (2S)-2-[4-[[[(6R)-2-amino-5-fonnyl-4-oxo-I,4,5,6.7,8-
H,N N
H
N
H ,
hexabydroptetidin-6-yl]methyl]amino]
benzamido]pentanedioic acid (dextrofolinic acid),
B. (2S)-2-[4-[[[(6R)-2-amino-5-fonnyl-4-oxo-I,4,5,6.7,8-
a H C~H
hexabydropteridin-6-yl]methyl)(fonnyl)amino]benzamido]
pentanedioic acid (5,1O-difonnyltetrahydrolevofolic acid), o ~~~C~H
H,N---{
N}
j-X JJ r- N
HN ,
HN ·'H
I. (2S)-2-[ 4-[(6aR)-3-amino-l-oxo-I,2,5,6,6a.7-
hexabydroimidazo[I,5-j]pteridin-8(9H)-yl]benzamido]
pentanedioic acid «6aR)-5,10-methylenetetrabydrofolic
C. (2S)-2-[4-[[(2-amino-4-oxo-I,4-<1ihydropteridin-6-yl) acid).
methyl]amino]benzamido]pentanedioic acid (folic acid). _________ ~ ""E"
Calcium Levulinate Dihydrate

D. (2S)-2-[4-[[(2-amino-4-oxo-1 ,4-<1ihydropteridin-6-yl)
methyl](fonnyl)amino]benzamido]pentanedioic acid (10-
Calf"
[ H'C ~CO"]
II
o 2
formylfolic acid),
C",!fI4CaO,,2H20 306.3 5743-49-7
Action and use

Source of calcium.
""E" _
DEFINITION
E. 4-[[[(6S)-2-amino-5-fonnyl-4-oxo-l,4,5,6.7,8- Calcium di(4-oxopentanoate) dihydrate.
hexabydropteridin-6-yl]methyl]amino]benzoic acid «6S)- Content
5-fonnyltetrabydropteroic acid), 98.0 per cent to 101.0 per cent (dried substance).
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1-410 Calcium Pantothenate 2022
CHARACTERS Chlorides (2.4.4)

Appearance Maximum 50 ppm.
WWte or almostwhite, crystalline powder. Dilute 10 mL of solution S [0 15 mL with waterR.
Solubility Sulfates (2.4.13)
Freelysoluble in water, very slightly soluble in ethanol Maximum 200 ppm.
(96 per cent), practically insoluble in methylene chloride. Dilute 7.5 mL of solution S to 15 mL with disnned waterR.
IDENTIFICATION Magnesium and alkali metals
First identification: A, D, H. Maximum 1.0 per cent.
Second identification: B, OJ D, E. To 10 mL of solution S, add 80 mL of water R, 10 mL of
A. Infrared absorption spectrophotometry (2.2.24). ammonium chloride solUIUm Rand 1 mL of ammonia R. Heat
Comparison calcium levulinate dihydrate CRS. to boiling. To the boiling solution, add dropwise 50 mL of
wann ammonium oxalate solution R. Allow to standfor 4 h,
then dilute to 200 mL with waterR and filter. To 100 mL of
Test solution Dissolve 60 mg of the substance to be the filtrate, add 0.5 mL of sulfun·c acidR. Evaporate to
examined in waler R and dilute to 1 mL with the same dryness on a water-bath and ignite to constant mass at
solvent. 600 ± 50 'C. The residue weighs a maximum of 5.0 mg.
Reference solution Dissolve 60 mg of calcium levulinate Loss on drying (2.2.32)
dihydrate CRS in waterR and dilute to I mL with the same 11.0 per cent to 12.5 per cent, determined on 0.200 g by
solvent. drying at 105 °C.
Plate TLC silica gelplate R.
Pyrogens (2.6.8)
Mobile phase concentrated ammonia R, ethyl acetate R, If intended for use in the manufacture of parenteral
waterR. ethanol (96 perem\! R (10:10:30:50 VIVIV/l-). preparations without a further appropriate procedure for the
Applicatiun I0 ~L. removal of pyrogens, it complies with the test for pyrogens.
Development Over a path of 10 cm. Injectper kilogram of the rabbit's mass 4 mL of a solution
Drying At 100-105 "C for 20 min and allow to cool. containing per millilitre 50 mg of the substance [Q be
examined.
Detection Spray with a 30 gIL solution of potassium
permanganate R. Dey in a current of warm airfor about ASSAY
5 min or until the spots become yellow. Examine in daylight. Dissolve 0.240 g in 50 mL of waterR. Carry out the
Results The principal spot in the chromatogram obtained complexometric titration of calcium (2.5.11).
with the test solution is similar in position, colourand size to I mL of 0.1 M sadium edetate is equivalent to 27.03 mg
the principal spot in the chromatogram obtained with the of C,oMI.CaO,.
reference solution. STORAGE
C. To I mL of solution S (see Tests), add 20 mL of a Protected from light.
2.5 gIL solution of dinitropheny/hydrazine R in dilute _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
hydroch/ori< aeid R. Allow to stand for 15 min. Filter, wash
the precipitate with water R. Dry the precipitate in an oven at
100-105 'C. The melting point (2.2.14) is 203 "C to 210 'C.
D. It gives reaction (b) of calcium (2.3.1).
E. Loss on drying (see Tests).
Calcium Pantothenate
TESTS (Ph. Bur. monograph 0470)
Solution S
Dissolve 10.0 g in carbon dioxide-free water R prepared from. H,C CH,O ]
disnned waterR and dilute to 100.0 mL with the same Ca2+ HO~ ~C0:2'
solvent.
[ H OH ~ 2
Solution S is clear (2.2.1) and not more intensely coloured 476.5 137-08-6
than reference solution Y, (2.2.2, Methad II).
pH (2.2.3) Action and use
6.8 to 7.8 for solution S. Componentof vitamin B.
Oxidisable substances PhE" _
To I mL of solution S, add 10 mL of waterR, I mL of dilute DEFINITION
su/furi< acidRand 0.25 mL of a 3.0 gIL solution of potassium Calcium bis[3-[(2R)-2,4-dihydroxy-3,3-dimethylbutanamido]
pennanganate R. Mix. After 5 min, the violetcolourof the propanoate] .
mixture is still visible.
Content
98.0 per cent 10 101.0 per cent (dried substance).
To 5 mL of solution S add 2 mL of hydrodllori< qcid RI and
dilute to 10 mL with waterR. Heat to boiling for 5 min and CHARACTERS
allow to cool. Add 10 mL of sodium carbonate soluutm R. Appearance
AUow to stand for 5 min, dilute to 25 mL with water R and White or almost white powder, slightly hygroscopic.
filter. To 5 mL of the filtrate add 2 mL of cupri-umasic Solubility
solution R and heat to boiling for I min. No red precipitate is Freely soluble in water, slightly soluble in ethanol
formed. (96 per cent) and practically insoluble in heptane.
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2022 Calcium Pantothenate 1-411
IDENTIFICATION Reference solution (c) Dissolve 3.0 mg of pomotactone CRS

First identification: A, B, D. (impurity C) in water R, add 1.0 mL of reference solution (a)
Second identification: C, D. and dilute to 100.0 mL with the same solvent.
A. Specific optical rotation (see Tests). Reference sohuion (d) Dissolve 3 mg of pantothenate for peak
B. Infrared absorption spectrophotometry (2.2.24). identification CRS (containing impurities B, E and H) in
0.5 mL of water R.
Comparison calcium pantothenate CRS.
Column:
C. Thin-layer chromatography (2.2.27). - size: 1= 0.15 m, '1 = 3.0 mntj
Test solution Dissolve 10 mg of the substance to be - statianary phase: actadecylsilyl silica gelfar chromatography R
examined in a mixture of 0.25 mL of water R and 4 mL of (3.5 pm);
methanol R. - temperature: 35 °C.
Reference solution Dissolve 10 mg of calcium Mabile phase:
pantothenate CRS in a mixture of 0.25 mL of waterR and - mobile phaseA: mix 1 volume of acetonitrile RJ and
4 mL of methanal R. 99 volumes of a 1.56 gIL solution of sodium dihydrogen
Plare TLC silica gelplare R. phosphare R previously adjusted to pH 2.5 with phospharic
Mobile phase glacial acetic acid R, Waler R, 2-propanol R ocidR;
(5:15:80 VIVIV). - mobile phase B: aceumiuile RJ;
Applicatlon 5 ~L.
Development Over 4/5 of the plate. (mln) (per cent I'm (per cent I'm
Drying In air. 0-. 100 0
Detection Heat at 120°C for 20 min; treat the warm plate 6·21 100 --> 50 0-->50
with a 3 gIL solution of ninhydrin R in a mixture of 21 - 30 50 50
3 volumes of glacial acetic acid Rand 100 volumes of
anhydrous ethanol R; allow to dry and heat again at 120°C Flow rate 1.0 mUmin.
for a few minutes; examine in daylight. Detection Spectrophotometer at 200 hID.
Results The principal spot in the chromatogram obtained Injection 5 JlL of the test solution and reference
with the test solution is similar in position, colourand size to solutions (b), (c) and (d).
Identification of impun·t1"es Use the chromatogram obtained
reference solution.
D. It gives reaction (a) of calcium (2.3.1). impurity C; use the chromatogram supplied with pantothenate
TESTS far peak identification CRS and the chromatogram obtained
Solution S with reference solution (d) to identify the peaksdue to
Dissolve 2.50 g in carbon dioxide-free waterR and dilute to impurities B, E and H.
50.0 mL with the same solvent. Relative retention With reference to calcium pantothenate
Appearance of solution (retention time = about 4 min): impurity B = about O.5j
Solution S is clear (2.2.1) and colourless (2.2.2, Melhod If). impurity C = about 0.8; impurity E = about 1.7;
pH (2.2.3)
=
- resohuion: minimum 3.0 between the peaks due to
Specific optical rotation (2.2.7) impurity C and calcium pantothenate;
+ 25.5 to + 27.5 (dried substance), determined on - signal-to-noise ratio: minimum 10 for the peak due to
solution S. impurity C.
Impurity A and otlter amlnocarboxylic acid impurities Cakuknion of percentage contents:
Maximum 0.50 per cent. - for impurities Band C, use the concentration of
Dissolve 8.000 g in 40 mL of waterR and dilute to 100 mL impurity C in reference solution (c);
with the same solvent. Add 25 mL of farmaldehyde tdudon R. - for impurities other than Band C, use the concentration
Titrate with 0./ M sodium hydroxide, determining the of calcium pantothenate in reference solution (b).
end-point potentiometricaUy (2.2.21J). Limits:
1 mL of 0.1 M sadium hydroxide is equivalent to 8.91 mg of - ;mpun·ty B: maximwn 0.8 per cent;
C,H7N 0 2 • - impurity C: maximum 0.3 per cent;
Related substances - impurity E: maximum 0.25 per cent;
Liquid chromatography (2.2.29). - impun·ty H: maximwn 0.15 per cent;
- unspecified impuriues: for each impurity, maximum
0.10 per cent;
solvent.
- reporting threshold: 0.05 per cent, disregard any peak due
Reference solution (aJ Dissolve 30.0 mg of calcium to impurity A (eluting before 1 min).
pantothenate CRS in water R and dilute to 50.0 mL with the
Chlorides (2.4.4)
same solvent.
Maximum 200 ppm.
100.0 mL with warer R. Dilute 1.0 mL of this solution to
Dilute 5 mL of solution S [0 15 mL with water R.
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1-412 Calcium Phosphate 2022

an oven at 105 DC.
ASSAY
Dissolve 0.180 g in 50 mL of anhydrous acetic acid R. Titrate H.3-[(53)-5-(I-hydroxy-2-methylpropan-2-yl)-4-oxo-l,3-
with 0.1 M perchknic acid, determining the end-point oxazolidin-3-yl]propanoic acid.
potentiometrically (2.2.20). _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE",
I mL of 0.1 M perchloric acid is equivalent '0 23.83 mg of
ClsH32CaN2010'
STORAGE
In an airtight container. Calcium Phosphate
IMPURlTIES Tribasic Calcium Phosphate
Specified impw;u'es A.. B, C, E, H.
Other detectable impuri,ies (thefollowing substances would, if
present at a sufficient level, be deteaed by oneor other of the tests Action and use
in the monograph. They are limited by the general acceptance Excipient.
cnierion for otherlunspeaJied impuniies andior by the general Preparations
monograph Subsumces for pharmaceuti<al use (2034). II is Calcium and Ergocalciferol Tablets
there/ore not necessary to idenufy these ;mpun"ties jor
Cal~um Phosphate for Homoeopathic Preparations
demonstration oj compliance; See also 5.10. Control oj impun'ties
in substanus for pharmaceutical use) D, F, G. Calcium and Ergocalciferol Chewable Tablets
PhE", _
DEFINITION
A. 3-aminopropanoic acid (p-alanine), Mixture of calcium phosphates.
Content
35.0 per cent '0 40.0 per cent of Ca (A, 40.08).
CHARACTERS
Appearance
Whiteor almost white powder.
B. (2R)-2,4-dihydroxy-3,3-dimethylbutanoic acid (pantoic
acid), Solubility
Practically insoluble in water. It dissolves in dilute
hydrochloric acid and in dilute nitric acid.
IDENTIFICATION
A. Dissolve0.1 g in 5 mL of a 25 per cent VIV solution of
nim'c acid R. The solution gives reaction (b) of phosphates
(2.3.1).
C. (3R)-3-hydroxy-4,4-dimethyloxolan-2-one (pantolactone), B. It gives reaction (b) of calcium (2.3.1). Filter before
adding potassium femxyanide sdution R.
C. It complies with me limits of the assay.
TESTS
Solution S
D. methyl 3-[(2R)-2,4-dihydsoxy-3,3-dimethylbutanamido] Dissolve 2.50 g in 20 mL of dilure hydrochloric acidR. If the
propanoate (methyl pantothenate), solution is not clear, filter it Add dilute ammonia RJ dropwise
until a precipitate is formed. Dissolvethe precipitate by
adding dilure hydrochloric add R and dilute to 50 mL with
distiUed waterR.
Chlorides (2.4.4)
Maximum 0.15 per cent:
E. 3-[3-[(2R)-2,4-dihydsoxy-3,3-dimethylbutanamido] Dissolve0.22 g in a mixture of I mL of nitric acid Rand
propanamido]propanoic acid (~-alanyl pantothenamide}, 10 mL of waterR and dilute to 100 mL with water R.
Fluorides
Maximum 75 ppm.
Porenticmetry (2.2.36, Method II).
F. 3,3'-azanediyldipropanoic acid, Test solution Dissolve 0.250 g in 0.1 M hydrochloric add, add
5.0 mL ofjluoride standard solution (J ppm F) R and dilute to
50.0 mL with 0.1 M hydrochloric acid, To 20.0 mL of this
solution add 20.0 mL of total-ionic-strength-adjustment buffer R
and 3 mL of an 82 gIL solution of anhydrous sodium
acetate R. Adjust to pH 5.2 with ammonia R anddilute lo
G. 3-(3-aminopropanamido)propanoic acid (Il-alanyl-~
50.0 mL with distilled waterR.
alanine),
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2022 Calcium Polystyrene Sulfonate 1-413
Reference solution Fluoride standard solution (10 ppm F) R.

Calcium Polystyrene Sulfonate
lndiauor electrode Fluoride-selective.
Calcium Polystyrene Sulphonate
Action and use
Carry out me measurements on the test solution, then add at
Used in the treatment of hyperkalaemia.
least 3 quantkles, each of 0.5 mI..., of the reference solution,
carrying out a measurement after each addition. Calculate DEFINITION
the concentrationof fluorides using the calibration curve, Calcium Polystyrene Sulfonate is a cation-exchange resin
taking into account the addition of fluoride to the test
prepared in the calcium fonn containing not less than
solution. 6.5% wlw and not more than 9.5% w/w of calcium,
Sulfates (2.4.13) calculated with reference to the dried substance. Each g
Maximum 0.5 per cent. exchanges not less than 1.3 mEq and not more than 2.0
Dilute I mL of solution S to 25 mL with d~ti/Ied water R. mEq of potassium) calculated with reference to the dried
Arsenic (2.4.2, Method A) substance.
Maximum 4 ppm, determined on 5 mL of solution S. CHARACTERISTICS
Iron (2.4.9) A cream to light brown, fine powder.
Maximum 400 ppm. Practically insoluble in water and in ethanol (96%).
Dilute 0.5 mL of solution S to 10 mL with water R. IDENTIFICATION
Acid-insoluble matter A. The infrared absorption spearum, Appendix IT A, is
Maximum 0.2 per cent. concordant with the reference spectrum of calcium polystyrene
Dissolve 5.0 g in a mixture of 10 mL of hydrochloric acid R sulfonate (RS 037).
and 30 mL of waler R. Filter, wash the residue with water R B. Yields reaction C characteristic of calcium salts,
and dry to constant mass at 100-105 °C. The residue weighs Appendix VI.
a-maximum of 10 mg.
TESTS
Loss on Ignidon Particle size
Maximum 8.0 per cent, determined on 1.000 g by ignition at Not more than 1% w/w is retained on a I50-p.m sieve,
800 ± 50°C for 30 min. Appendix XVII B. Use 20 g and sieve for 5 minutes.
ASSAY Potassium
Dissolve 0.200 g in a mixture of I mL of hydrochloric acid R/ Not more than 0.1 % of K when determined by atomic
and 5 mL of waterR. Add 25.0 mL of 0./ M sodium edetate emission spectrophotometry, Appendix II D, measuring at
and dilute to 200 mL with water R. Adjust to about pH 10 766.5 run and using a solution prepared in the following
with concenuated ammonia R. Add 10 mL of ammonium manner. To 1.1 g of the substance being examined add
chloride buffer sdtuion pH / O. 0 R and a few milligrams of 5 mL of hydrochloric acid, heat to boiling, cool and add
mordant black 11 triturate R. Titrate the excess sodium edetate 10 mL of water. Filter, wash the filter and residue with waler
with 0.1 M zinc sulfate until the colour changes from blue to and dilute the filtrate and washings to 25 mL with water.
violet. Use potassium standard solution (100 ppm K), suitably diluted
l mL of 0.1 kI sodium edetate is equivalent to 4.008 mg of with water, to prepare the standard solutions.
Ca. Sodium
FUNCTIONALITY-RELATED CHARACTERISTICS Not more than 0.1 % ofNa when determined by atomic
This section provides infonnation on characteristics that are emission spectrophotometry, Appendix ITD, measuring at
recognised as being relevant control parameters for oneor more 589.0 urn and using a solution prepared in the following
manner. To 1.1 g of the substance being examined add
junctions of the substance when usedas an excipient (see chapter
5 mL of hydrochloric acid, heat to boiling, cool and add
5.15). Some of the characteristics described in the FuncUcnalily-
10 mL of water. Filter, wash the filter and residue with water
part of the monograph since they also represent mandatory quality and dilute the filtrate and washings to 25 mL with water.
criteria. In such cases) a cross-reference to the tests described in the Use sodium so/utinn (200 ppm Na), suitably diluted with
mandatory part is included in the Functionality-related waur, to prepare the standard solutions.
characutistics section. Control of the characteristics can contribute Arsenic
UJ the quality of a medicinal product by improving the consistency 1 g dispersed in 25 mL of water complies with the limit test
of the manufacturing process and the petformance of the medicinal for arsenic, Appendix vn (I ppm).
product during use. Where control methods are cited, they are Styrene
recognised as being suitable for em purpose, but other methods can Carry out the method for liquidchromatography,
also be used. Wherever results for a particular characteristic are Appendix III D, using the following solutions.
reponed) the contrd methodmust be indicated. (I) Shake 10 g of the substance being examined with 10 mL
The following characteristics may be relevant for calcium of acetone for 30 minutes, centrifuge and use the supernatant
phosphate is usedas a filler in tablets and capsules. liquid.
Particle-size distribudon (2.9.3/ or 2.9.31f) (2) 0.0001 % wlv of styrene in acetone.
Bulk and tapped density (2.9.34) CHROMATOGRAPHIC CONDITIONS
Powder flow (2.9.36) (a) Use a stainless steel column (30 em x 4 mm) packed
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" with O<tadecylsi/yl silica gelfor chromatography CltBondapak
CI8 issuitable).
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1-414 Calcium Stearate 2022
(b) Use isocratic elution and the mobile phase described

below.
Calcium Stearate
(e) Use a flow rate of 2 mL per minute. (Ph. Eur. monograph 0882)
(d) Use an ambient column temperature.
1592-23-0
(e) Use a detection wavelength of 254 nm.
(f) Inject 20 pL of each solution. Action and use
Excipient.
MOBILE PHASE
PhE<r _
Equal volumes of acetonitrile and water.
LIMITS DEFINITION
In me chromatogram obtained with solution (1): Mixture of calcium salts of dilTerent fatty acids consisting
the area of any peak corresponding to styrene is not greater mainly of stearic (octadecanoic) acid [(C. 7H"COO),Ca;
than the area of the peak in me chromatogram obtained with Me- 607] and palmitic (hexadecanoic) acid
solution (2) (I ppm). [(C 15HJlCOO),Ca; M, 550.9) with minor proportions of
other fatty acids.
Potassium exchange capacity
To 3 g of the substance being examined in a dry 250 mL Content
glass-stoppered flask add 100 mL of a solution containing - calcium: 6.4 per cent to 7.4 per cent (A, 40.08) (dried
0.7455% wlv of porassium chloride and 0.4401% wlv of substance);
potassium hydrogen carbonate in-water (solution A), stopper - stearic acid in thefauy acidfraction: minimum
and shake for 15 minutes. Filter and dilute 2 mL of the 40.0 per cent;
filtrate to 1000 mL with water. Determine the concentration - sum of stearic acid and palmitic acid in thefatty acid fraction:
of unbound potassium in this solution by atomic emission minimum 90.0 per cent.
spectrophorcmetry, Appendix II D, measuring at 766.5 nm and CHARACTERS
using solution A suitably diluted with water, to prepare the Appearance
standard solutions. Calculate the potassium exchange Fine, white or almost white, crystalline powder.
capacity of the substance being examined in milliequivalents
Solubility
taking the concentration of potassium in solution A as
Practically insoluble in water and in ethanol (96 per cent).
144 milliequivalents of K per litre.
IDENTIFICATION
Loss on drying
When dried at 70° at a pressure not exceeding 0.7 kPa for First identification: C, D.
16 hours, loses not more than 8.0% of its weight. Use 2 g. Second identification: A, H, D.
Microbial contamination A. Freezing point (2.2.18): minimum 53 °C J for the residue
Carry out a quantitative evaluation for Enterobacteria and obtained in the preparation of solution S (see Tests).
certain other Gram-negative bacteria, Appendix XVI BI. B. Acid value (2.5./): 195 to 210.
0.01 g of the substance being examined gives a negative Dissolve 0.200 g of the residue obtained in the preparation of
result, Table I (most probable number of bacteria per gram solution S in 25 mL of the prescribed mixture of solvents.
fewer than 10'). C. Examine the chromatograms obtained in the test for fatty
ASSAY acid composition.
For calcium Results The retention times of the principal peaks in the
Carefully heat 1 g in a platinum crucible until a white ash is chromatogram obtained with the test solution are
obtained and dissolve in 10 mL of 2M hydrochloric acid with approximately the same as those of the principal peaks in the
the aid of heat. Transfer the resulting solution to a conical chromatogram obtained with the reference solution.
llask using 20 mL of water. Add 50 mL of 0.05Mdisodium D. Neutralise 5 mL of solution S to redlitmus paperR using
edeuue VS, 20 mL of ammonia buffer pH 10.9 and titrate the strong sodium hydroxide solution R. The solution gives
excess of disodium edetate with O.02M zinc sulfate VS, using a reaction (b) of calcium (2.3.1).
0.5% wlv solution of mordant black 11 in ethanol (96%) as
indicator to a red purple end point. Each mL of O.05M TESTS
disodium edeuue VS is equivalent to 2.004 mg of Ca. Solution S
To 5.0 g add 50 mL of peroxide-free ether R, 20 mL of dilul<
STORAGE nitric acid Rand 20 mL of distilled water R. Boil under a
Calcium Polystyrene Sulfonate should be kept in an airtight reft.ux condenser until dissolution is complete. Allow to cool.
container. In a separating funnel, separate the aqueous layer and shake
the ether layer with 2 quantities, each of 5 mI.., of distilled
waterR. Combine the aqueous layers, wash with 15 mL of
peroxide-free ether R and dilute the aqueous layer to 50 mL
with distilled water R (solution S). Evaporate the ether layer to
dryness and dry the residue at 100-105 °C. Keep the residue
for identification tests A and B.
To 1.0 g add 20 mL of carbon dioxide-free water R and boil
for 1 min with continuous shaking. Cool and filter.
To 10 mL of the filtrate add 0.05 mL of bromot!lymol blue
solutio.. RI. Not more than 0.5 mL of 0.01 M hydrothloric
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2022 Dried Calcium Sulfate 1-415
"'I
add or 0.01 sodium hydroxide is required to change the Detection Flame ionisation.
colour of the indicator. Injection I ~L.
Chlorides (2.4.4) Relative retention With reference to methyl stearate: methyl
Maximum 0.1 per cent. palmitate = about 0.9.
Dilute 0.5 mL of solution S to 15 mL with water R. System suitability Reference solution:
Sulfates (2.4.13) - resolution: minimum 5.0 between the peaks due to methyl
Maximum 0.3 per cent. palmitate and methyl stearate.
Dilute 0.5 mL of solution S to 15 mL with distiUed waterR. Calculate the content of palmitic acid and stearic acid.
Loss on drying (2.2.32) FUNCTIONALITY-RELATED CHARACTERISTICS
Maximum 6.0 per cent, determined on 1.000 g by drying in This section provides infonnatian on charaaetistics that are
an oven at 105 "C. rec-ognised as being relevam control parameters for one or more
Microbial contamination functions of the subsumce when used as an exdpient (see chapter
TAMC: acceptance criterion 103 CFU/g (2.6.12). 5.15). Someof the characteristics described in the Funaionalil~
related characteristics section may also be present in the mandatory
pan of the monograph since they also represent mandatory quality
Absence of Escherichia cali (2.6.11). criteria. In such cases, a cross-reference to the tests described in the
Absence of Salmonella (2.6.11). mandatory pan is included in the Functionality-related
ASSAY characteristics section. Control of the charaaetistiacan contribute
Calcium to the quality of a medicinal product by improving the consistency
To 0.500 g in a 250 mL conical flask add 50 mL of a of the manufacturing process and the peifonnance of the medicinal
mixture of equal volumes of anhydrous ethanol Rand product during use. W'here control methods arecited, they are
butanol R, 5 mL of concentrated ammonia R, 3 mL of recognised as being suitable for the purpose, but othermethods can
ammonium chloride buffersolutian pH 10.0 R, 30.0 mL of also be used. Wherever results for a paru'cular characteristic are
0.1 M sodium edetate and 15 mg of mordant black 11 reported, the control methodmust be indicated.
triturate R. Heat to 45-50 °C until the solution is clear. Cool Thefollowing characteristics may be relevant for calcium stearate
and titrate with 0.1 M zinc sulfate until the colour changes usedas a lubricant in tablets and capsules.
from blue to violet. Carry out a blank titration. Particle-size distribution (2.9.31)
1 mL of 0.1 M sodium edetate is equivalent to 4.008 mgcf Specific surface area (2.9.26, Method 1)
Ca. Determine the specific surface area in the PIPo range of
Composition of fatty acids 0.05 to 0.15.
Gas chromatography (2.2.28): use the normalisarion Sample outgassing 2 h at 40 "C.
procedure. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>EII
Testsolution In a conical flask fitted with a reflux condenser,
dissolve 0.10 g of the substance to be examined in 5 mL of
boron trifluoride-methanol solution R. Boil under a reflux
condenser for 10 min. Add 4 mL of heptane R through the
condenser. Boil under a reflux condenser for 10 min. Allow
Dried Calcium Sulfate
to coo!' Add 20 mL of saturated sodium chloride solution R. Exsiccated Calcium Sulfate; Plaster of Paris; Dried Calcium
Shake and allow the layers to separate. Remove about 2 mL Sulphate
of the organic layer and dry over 0.2 g of anhydrous sodium CaSO,,'hH,O 145.1 26499-65-0
suifaUi R. Dilute 1.0 mL of the solution to 10.0 mL with
DEFINITION
heptane R.
Dried Calcium Sulfate is prepared by heating powdered
Reference solution Prepare the reference solution in the same gypsum, CaS04.l2H20, at about 1500 in a controlled manner
manner as the test solution using 50.0 mg of palmitic such that it is substantially converted into the hemihydrate,
acidCRS and 50.0 mg of ,rearic acidCRS instead of calcium CaS04,JhH 20, with minimum production of the anhydrous
stearate. phases of calcium sulfate. It may contain suitable setting
Column: accelerators or decelerators.
CHARACTERISTICS
=
- size: / ;;:; 30 m, 0 0.32 nun;
A white or almost white powder; hygroscopic.
- s"1tionary phase: macrogol 20 000 R (film thickness
0.5 pm). Slightly soluble in water; more soluble in dilute mineral acids;
practically insoluble in ethanol (96%).
Gamer gas helium for chromatography R.
Flow rate 2.4 mUmin. IDENTIFICATION
Temperature: Yields the reactions characteristic of calcium salts and of
sul/ates, Appendix VI.
Time Temperature TESTS
(min) CC) Setting properties
Column 0-2 7. 20 g mixed with 10 mL of water at 15" to 20" in a cylindrical
2·36 70 ...... 240 mould about 2.4 cm in diameter sets in 4 to 11 minutes.
36 - 41 '4. The mass thus produced, after standing for 3 hours,
Injection port 220 possesses sufficient hardness to resist pressure of the fingers
Detector '60 at the edges, which retain their sharpness of outline and do
not crumble.
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1-416 Calcium Sulfate 2022
Loss on ignition FUNCTIONALITY-RELATED CHARACTERISTICS

When ignited to constant weight at red heat, loses 4.5% to This section provides infonnation on characteristics that are
8.0% of its weight Use 1 g. recognised as being relevant control parameters for oneor more
functions of the substance tshen usedas an excipiem (see chapter
5.15). Some 0/the characteristics described in the Functionality-
related charaaetistics section may alsobe present in the mandatory
Calcium Sulfate Dihydrate part of the monograph since they also represent mandatory quality
cruetia. In such cases, a cross-reference to the tests described in the
Calcium Sulphate mandasoty pan is induded in the Funetwnality-relaud
(ph. Bur. monograph 0982) characteristics section. Control of the characrnistics Can contribute
ro the quality of a medicinal produce by improving the consistency
CaSO,,2H,O 172.2 JOJOJ-4J-4
of the manufacturing process and theperformance of the medidnal
Acdon and use product during use. W'here control methods are cited, they are
Excipient recognised as being suitable for thepurpose, but other methods can
"'Eos ~ ~ _ reported, the control methodmust be indicated"
DEFINITION Thefollowing characteriuia may be relevant for calcium sulfate
Content dihydrate usedas filler in tablets and cap,ules.
98.0 per cent to 102.0 per cent of CaSO.,,2H,O. Particle-size distribution (2.9.3J or 2.9.38)
CHARACTERS Bulk and tapped density (2.9.34)
Appearance Powder flow (2.9.36)
Whiteor almost white fine powder. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "'E",
Solubility
Very slightly soluble in water, practically insoluble in ethanol
(96 per cent).
Natural Camphor ***
IDENTIFICATION *** ***
A. Loss on ignition (see Tests). (~Camphor, Ph. Eur. monograph 1400) ***
B. Solution S (see Tests) gives reaction (a) of sulfates (2.3. J).
C. Solution S gives reectionta) of calcium (2.3.J). CH,
! 0
TESTS H3C.r1"'f
Solution S H,c'V
Dissolve 1.0 g in 50 mL of a 10 per cent VIV solution of
hydrochlori< acidR by heating at 50 'C for 5 min. Allow to ~
cool.
Acidity or alkalinity 152.2 464-49-3
Shake 1.5 g with 15 mL of carbon du,xide-jree water R for "'Eos _
5 mio. Allow to stand for 5 mio and filter. To 10 mL of the DEFINITION
filtrate add 0.1 mL of phenolphthalein ,oI",u,n Rand 0.25 mL
(I R,4R)-I,7,7 -Trimethylbicyclo[2.2.1]heptan-2-one.
of aOJ M sodium hydroxide. The solution is red.
Add 0.30 mL of 0.01 M hydrochloric acid. The solution is CHARACTERS
colourless. Add 0.2 mL of methyl red,olu"on R. The solution Appearance
is reddish-orange. Whiteor almost white, crystalline powder or friable,
Chlorides (2.4.4) crystalline masses.
Maximum 300 ppm. HigWy volatile even at room temperature.
Shake 0.5 g with 15 mL of water R for 5 min. Allow to stand Solubility
for 15 min and filter. Dilute 5 mL of the filtrate to 15 mL Slightly soluble in water, very soluble in alcohol and in light
with water R. petroleum, freely soluble in fatty oils, very slightly soluble in
Iron (2.4.9) glycerol.
Maximum 100 ppm. IDENTIFICATION
To 0.25 g add a mixture of 5 mL of hydrochlori< acidR and First identification: A, c.
20 mL of water R. Heat to boiling, cool and filter. Second idendficauon: A, B, D.
Loss on Ignition A. Specific optical rotation (see Tests).
18.0 per cent to 22.0 per cent, determioed on 1.000 g by B. Melting point (2.2.J4): 175 'C to 179 'C.
ignition to constant mass at 800 ± 50 DC.
ASSAY Comparison racemic camphor CRS.
Dissolve 0.150 g in 120 mL of water R. Carry out the D. Dissolve 1.0 gin 30 mL of methanol R. Add 1.0 g of
complexometric titration of calcium (2.5. lJ). hydroxylamine hydrochloride Rand 1.0 g of anhydrous sodium
1 mL of a1 1"1 sodium edetate is equivalent to 17.22 mg acerate R. Boil undera reflux condenser for 2 h. Allow to
of CaSO,,2H,O. cool and add 100 mL of water R. Filter, wash theprecipitate
obtained with 10 mL of water Rand recrystallise from 10 mL
of a mixture of 4 volumes of alcohol Rand 6 volumes of
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2022 Camphor 1-417
water R. The crystals, dried in vacuo, melt (2.2.14) at 118 "C - totalof other impurities: not more than 4 times the area of
(0 121 -c, the principal peak in the chromatogram obtained with
TESTS
Cany out the weighings and dissolution rapidly.
Solution S (0.05 per cent).
Dissolve 2.50 g in 10 mL of alcohol R and dilute to 25.0 mL
Halogens
Maximum 100 ppm.
Dissolve 1.0 g in 10 mL of Z-propatlol R in a distiUation flask.
Solution S is clear (Z.Z.1) and colourless (Z.Z.Z, Method11).
Add 1.5 mL of dilute sodium hydroxide sdution Rand 50 mg
Acidity or alkalinity of nichel-aluminium alloy R. Heat on a water-bath until the
To 10 mL of solution S add 0.1 mL of phenolphthalein Z-propanol R has evaporated. Allow (0 cool and add 5 mL of
solution R 1. The solution is colourless. Not more than water R. Mix and filter through a wet filter previously washed
0.2 mL of 0.11\11 sodium hydroxide is required to change the with water R until fiee from chlorides. Dilute the filtrate to
colour of the indicator. 10.0 mL with waler R. To 5.0 mL of the solution, add n'-m·e
Specific optical rotation (Z.Z.7) acid R dropwise until the precipitate which forms is
+ 41.0 to + 44.0, determined on solution S. redissolved and dilute to 15 mL with waterR. The solution
complies with the limit test for chlorides (2.4.4).
Related substances
Gas chromatography (Z.Z.ZIf). Residue on evaporation (2.8.9)
Test solution Dissolve 2.50 g of the substance to be Maximum 0.05 per cent.
examined in heptane R and dilute to 25.0 mL with the same Evaporate 2.0 g on a water-bath and dry in an oven at
solvent. 100-105 "C for I h. The residue weighs a maximum of
Reference ,olution (a) Dilute 1.0 mL of the test solution 10 I mg.
100.0 mL with heptane R. Water
Reference sohuion (b) Dilute 10.0 mL of reference Dissolve 1 gin 10 mL of light Pitroleum R. The solution is
solution (a) to 20.0 mL with heptane R. dear (Z.Z.l).
Reference solution (c) Dissolve 0.50 g of borneol R in IMPURITIES
heptane R and dilute to 25.0 mL with me same solvent.
H':~
Dilute 5.0 mL of the solution to 50.0 mL with heptane R.
Reference ,olution (d) Dissolve 50 mg of linalol Rand 50 mg and enantiomer
of bornyl acetate R in heptane R and dilute to 100.0 mL with
H3C ~
the same solvent. H
Column:
A. 2,6,6-trimethylbicyclo[3.I. l)hept-2-ene (c-pinene),
- size: I ;;;; 30 m, 0 ;;;; 0.25 1IlD1)
- stationary phase: macrogol ZO 000 R (0. 25 urn). H
Comer gas helium for chromatagraphy R.

Splli ratio 1:70.
ett-~~, and enanuomer
; CH2
Flow rate 45 cmls.
H
Temperature:
B. 2,2-dimethyl-3-methylenebicyclo[2.2.I]heptane
Tim. Temperature (camphene),
(min) Cq
Column 0-10 50
10 - 35 50 100
35·45 100 200
45 - 55 200
Injection port 220
Detector 250 C. 6,6-dimethyl-2-methylenebicydo[3.1.I)heptane
(p-pinene),
Detection Flame ionisation. CH,
Injection I ~L.
System suitability Reference solution (d).
- resolution: minimum 3.0 between the peaks due to bornyl tSCH'
CH,
acetate and to linalol.
Limits: D. 1,3,3-trimethyl-2-oxabicydo[2.2.2]octane (cineole),
- borneol: not more than the area of the principal peak in
(2.0 per cent),
- any other impurity: not more than half of the area of the
E. I,3,3-trimethylbicydo[2.2.!]heptan-2-one (fenchone),
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1-418 Camphor 2022
CH, PhEIl _
~:
~CH3
and onantiomer DEFINITION
(IRS,4RS)-I, 7,7 -Trimethylbicyclo[2.2.I)heplan-2-one.
I CH 3
H CHARACTERS
Appearance
F. em-I,3,3-trirnethylbicyclo[2.2.I)heplan-2-01 (fenchol), White or almost white, crystalline powderor friable,
crystalline masses, highly volatile even at room temperature.
H Solubility
~
; ,O~H, Slightly soluble in water, very soluble in ethanol
and enanUomer (96 per cent) and in light petroleum, freely soluble in fatty
CH,
oils, very slightly soluble in glycerol.
i CH3
H IDENTIFICATION
First idendfiouion: A, C.
G. ex<>-2,3,3-trirnethylbicyclo[2.2.I]heptan-2-01 (camphene Second identification: A, B, D.
hydrate),
A. Optical rotation (see Tests).
B. Melting point (2.2.14): 172 'C 10 180 'C.
Preparation Mulls in liquidparaffin R.
Comparison racemic camphor CRS.
D. Dissolve 1.0 g in 30 mL of methanol R. Add 1.0 g of
hydroxylamine hydrochloride Rand 1.0 g of anhydrous sodium
H. endo-2,3,3-trirnethylbicyclo[2.2 .I)heptan-2-ol acetate R. Boil undera reflux condenser for 2 h. Allow to
(methylcamphenilol), cool and add 100 mL of water R. A precipitate is formed.
Filter, wash with 10 mL of water Rand recrystaUise from
10 mL of a mixture of 4 volumes of ethanol (96 per cent) R
and 6 volumes of water R. The crystals, dried in vacuo) melt
(2.2.14) al118 -c 10 121 'C.
TESTS
Carryout the weighings rapidly.
1. eec-I,7, 7-trirnethylbicyclo[2.2.I)heplan-2-01 (exo-bomeol), Solution S
Dissolve 2.50 g in 10 mI. of ethanol (96 per cent) Rand
dilute [0 25.0 mL with the same solvent.
Acidity or alkal1nlty
Dissolve 1.0 g in 10 mL of ethanol (96 per cent) R and add
0.1 mL of phenolphthalein solution Rl. The solution is
J. endo-l,7, 7-trirnethylbicyclo[2.2.I]heptan-2-01 (ende- colourless. Not more than 0.2 mL of 0.1 M sodium hydroxide
borneol). is required to change the colour of the indicator.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph Ell Optical rotation (2.2.7)
-0.15° to + 0.15°, determined on solution S.
Related substances
Racemic Camphor Test sohuion Dissolve 50 mg of the substance to be
examined in hexane R and dilute to 50.0 mL with the same
(Ph. Bur. monograph 0655) solvent.
Reference solution (a) Dissolve 50 mg of the substance to be
examined and 50 mg of bornyl acetate R in hexane Rand
dilute to 50.0 mL with the same solvent.
and enanliomer
200.0 mL with hexane R.
Column:
- size: I :::: 2 ill) 0 :::: 2 mrn;
152.2 76-22-2 - stationary phase: diatomaceous earth for gas
chromatography R impregnated with 10 per cent m/m of
Action and use m""rogol 20 000 R.
Counter-irritant.
Carrier gas nitrogen for chromatography R.
Preparations
Flew rate 30 mUmin.
Camphorated Opium Tincture
Temperature:
Concentrated Camphorated Opium Tincture
- column: 130 "O,
Concentrated Camphor Water - injection pqrt and detector: 200°C.
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2022 Candesartan Cilexetil 1-419
Detection Flame ionisation. Content

Injection I ~L. 99.0 per cent to 101.0 per cent (anhydrous substance).
Run time 3 times the retention time of camphor. PRODUCTION
System $/Iimbility: As N-nitrosamines are classified as probable human
- resolution: minimum 1.5 between the peaks due to carcinogens, their presence in candesartan ciJexetil should be
camphor and bornyl acetate in the chromatogram avoided or limited as much as possible. For this reason,
obtained with reference solution (a); manufacturers of candesartan cilexetil for human use are
- signal-to-noise ratio: minimum 5 for the principal peak in expected to perform an assessment of the risk of
the chromatogram obtained with reference solution (b). N-nitrosamine formation and contamination during their
Limits: manufacturing process; if this assessment identifies a
- mOl impurity: for each impurity, not more than 2 per cent potential risk, the manufacturing process should be modified
of the area of the principal peak; to minimise contamination and a control strategy
- total: not more than 4 per cent of the area of the principal implemented to detect and control N-nitrosamine impurities
peak; in candesartan cilexetil. The general chapter 2.5.42.
- disregard limit: the area of the principal peak in the N-Nitrosamines in active substances is available to assist
chromatogram obtained with reference solution (b). manufacturers.
Halogens CHARACTERS
Dissolve 1.0 g in 10 mL of 2-propanol R in a distillation flask. White or almost white powder.
Add 1.5 mL of dilute sodium hydroxide solution Rand 50 mg Solubility
of nickel-aluminium allay R. Heat on a water-bath until the Practically insoluble in water, freely soluble in methylene
2-propanol R has evaporated. Allow to cool and add 5 mL of chloride and slightly soluble in anhydrous ethanol.
water R. Mix and filter through a wet filter previously washed
with waterR until free from chlorides. Dilute the filtrate to
10.0 mL with water R. To 5.0 mL of this solution, add niuic IDENTIFICATION
acid R dropwise until the precipitate which fonns is Infrared absorption spectrophotometry (2.2.24).
redissolved and dilute to 15 mL with water R. The solution Comparison candesanan ciJexeu1 CRS.
complies with the limit test for chlorides (2.4.4).
Water substance to be examined and the reference substance
Dissolve 1 gin 10 mL of light petroleum R. The solution is separately in anhydrous ethanol R, evaporate to dryness and
clear (2.2.1). . record new spectra using the residues.
TESTS
Related substances
Evaporate 2.0 g on a water-bath and dry at 100-J05 °C for
J h. The residue weighs not more man
1 mg.
_~ PhE"
Solvent mixture water R, acetonitrile R (40:60 VIV).
examined in 50.0 mL of the solvent mixture.
Candesartan Cilexetil Reference solution (a) Dilute 1.0 mL of the test solution to
100.0 mL with the soivent mixture. Dilute 1.0 mL of this
(ph. Bur. monograph 2573) solution to 10.0 mL with the solvent mixture.
Reference solution (b) Dissolve 5 mg of candesartan ciI"",ulfar
°
H CH,~\
J-o/"o
0 and enanliomer
system suitability CRS (containing impurities A, B andF) in
the solvent mixture and dilute to 10 mL with the solvent
mixture.
C(0?O",
H,t:"" Reference solution (c) Dissolve 2.5 mg of candesarum ciiexetil
for peak identification CRS (containing impurities G and H) in
the solvent mixture and dilute to 5 mL with the solvent
"~ A' mixture.
° ""'I
( CH, '" Column:
- size: 1= 0.15 m, 0 = 3.9 mm;
- stationary phase: end-capped oetadecylsilyl silica gelfar
611 145040-37-5 chromatography R (4 urn).
Mobile phase:
Action and use
- mobile phaseA: glacial acetic acidR, waterfor
Angiotensin II (AT!) receptor antagonist.
chromatography R. acetonitrile R (1:43:57 VIVIV);
Preparation - mobile phase B: glacial acetic acidR, waterfor
Candesartan Tablets chromatography R, acetonitrile R (1:10:90 VIVIV);
PhE" _
DEFINITION
(I RS) -i-[[(Cyclohexyloxy)carbonyl] oxy]ethyl z-ethoxv-t-
[[2 '-( I H- tetrazol-5-yl)biphenyl-4-yi] me!hyl]-i H-
benzimidazole-7-carboxylate.
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1-420 Candesartan Cilexetil 2022
Tim. MobUe phase A MObile phase B in the monograph. They are limited by the general a"eptance
(min) (per cent VjJJ) (per cent VIJI)
cruerion for other/unspecified impun'ties and/or by thegeneral
0-3 100 0 monograph Substances for pharmaceutical use (2034). I, is
3 - 33 100 -> 0 0--> 100 therefore not neussary to identify these impuniies for
33·40 0 100 demonstration of compliance. S~ also 5.10. Control of impuruies
in substances for pharmaceutical use) C, D, E, 1.
Injection 10 ~L
with candesartan cilexetil for system suitability CRS and the
identify the peaks due to impurities A, Band F; use the
chromatogram supplied with amdesanan tilexemfor peak
reference solution (c) to identify the peaks due £0 A. ethyI2-<:thoxy-l-[[2'-(IH-letrazol-5-yl)biphenyl-4-yl)
impurities G and H. methyl]-I H-benzimidazole-7-carboxylale,
Relative retention With reference to candesartan cilexetil
=
(retention time = about 11 min): impurity G about 0.2;
=
impurity A about 0.4; impurity B about 0.5; =
impurity F = about 2.0; impurity H = about 3.5.
HCH,}-
° oJ-o 0
0 and enaneorner
et~
-= N=N
impurities A and B. HN )'
N ""
Limits: HN~ .#,p-
- correction [acton: for the calculation of content, multiply O I
corresponding correction factor: impurities A and
'"
=
G 0.7; impurity H 1.6; = B. (IRSJ-I-[[(cyclohexyloxy)carbonyl]oxy]ethyl 2-oxo-3-[[2'-
- impurity B: not more than 3 times the area of the (I H-letrazol-5-yl)biphenyl-4-yl] methyl]-2,3-dihy dro-IH-
principal peak in the chromatogram obtained with benzimidazole-a-carboxylate,
- impurities F} G: for each impurity, not more than twice the
- impurities A, H: for each impurity) not more than
(0.15 per cent);
- unspecified impun"ties: for each impurity) not more than the
- uxal: not more than 6 times the area of the principal peak C. (IRSJ-I-[[( cyclohexyloxy)carbonyl]oxy]ethyl 3-[[2'-(1-
in the chromatogram obtained with reference solution (a) ethyl-I H-letrazol-5-yI) bipheny1-4-yl]methyl]-2-oxo-2,3-
(0.6 per cent); dihydro-lH-benzimidazole-4-carboxylate,
(0.05 per cent).
Water (2.5.32)
Maximum 0.3 per cent) determined on 60.0 mg.
ASSAY
Dissolve 0.500 g in 60 mL of glacial acetic acid R. Titrate
immediately with 0.1 M perchlotic acid) determining the
end-point potentiometrically (2.2.20) at the IS[ inflexion
point. D. (IRSJ-I-[[(cyclohexyloxy)carbonyl]oxy)ethyl 3-[[2'-(2-
ethyl-2H-letrazol-5-yl)biphenyl-4-yl]methyl]-2-oxo-2,3-
1 mL of 0.1 M perchlotic acid is equivalent tc 61.1 mg of
dihydro-IH-benzimidazole-4-carboxylate,
C"H,.,N.O•.
IMPURITIES
Specified impuruies A) B, F} G, H.
Other deltCtabie impurities (the following substances would, if
present at a sufficient level) be detected by oneor other of the tests
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2022 Capecitabine 1-421
0
H CH,~\or D
oJ--. 0 and enantlome,
Capecitabine
cl:t
~
~
#
N=(
N~N"
o
I
HI
c~ #
N=N
""I
( CH, '"
E. (lRSJ-I-[[(cyclohexyloxy)carbonyl]oxy]ethyl 2-ethoxy-l-
[[2'-(I-ethyl-I H- tetrazol-5 -yl)biphenyl-4-yl]methyl]-1H-
benzimidazole-7 -carboxylate,
359.3 /5436/-50-9
Action and use

Pyrimidine analogue; cytotoxic; treatment of colorectal
cancer.
Preparation
Capecitabine Tablets
PIlE" _
DEFINITION
F. (IRSJ-I-[[(cyclohexyloxy)carbonyl]oxy]ethyl 2-ethoxy-l- Pentyl [1-(5-deoxy-p-D-ribofuranosyl)-5-f1uoro-2-oxo-I,2-
[[2 '-(2-ethyl-2H-tetrazol-5-yl)biphenyl-4-yl]methyl]-I H- dihydropyrimidin-4-yl]carbamate.
benzimidazole-7-earhoxylate, Content
Co,H
QN~H,t:NN
CHARACTERS
Appearance
N=( 1#
o ""I Solubility
( CH, '" Sparingly soluble in water, freely soluble in anhydrous
ethanol, practically insoluble in heptane.
G. 2-ethoxy-l-[[2 '-( IH-tetrazol-5-yl)biphenyl-4-yl] methyl]- IDENTIFICATION
IH-benzimidazole-7-carboxylic acid (candesartan), A. Specific optical rotation (see Tests).
Comparison capecitabine CRS.
TESTS
Specific opdcal rotation (2.2.7)
+ 96.0 to + 100.0 (anhydrous substance).
the same solvent.
Related substances
immediately before use or store them at 2-8 "C;
H. (IRSJ-I-[[(cyclohexyloxy)carbonyIJoxy]ethyl 2-ethoxy-l- Solvent mixture acetonitrile R, methanol R, waterR
[[2'-[1-(triphenylmethyl)-I H-tetrazol-5-yl] biphenyl-4-yl] (5:35:60 VlViI').
methyl]-IH-benzimidazole-7-carboxylate (N- Testsolution Dissolve 60.0 mg of the substance to be
tritylcandesartan), examined in 80 mL of the solvent mixture, sonicate until
dissolution is complete and dilute to 100.0 mL with the
solvent mixture.
Reference solutian (aJ Dissolve 60.0 mg of capedcabine CRS
in 80 mL of the solvent mixture, sonicate until dissolution is
complete and dilute to 100.0 mL with the solvent mixture.
Reference solurion (c) Dissolve 3 mg of capecitabine
I. methyl 2-ethoxy-I-[[2'-(IH-tetrazol-5-yl)biphenyl-4-yl] impun'ty A CRS, 3 mg of capecitabine impurity B CRS and
methyl]-IH-benzimidazole-7-carboxylate. 5 mg of copedtabine impurity D GRSin 80 mL of the solvent
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<I
mixture, sonicate until dissolution is complete and dilute to
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1-422 Capecitabine 2022
100.0 mL with the solvent mixture. Dilute 1 mL of the in the monograph. They are limited by thegeneral acceptance
solution to 50 mL with the test solution. criurum for other/unspecified impuruies and/or by thegeneral
Column: monograph Substances for pharmaceutical use (2034). It is
- size: 1= 0.25 m, 0 ;::; 4.6 mm; therefore not necessary to identify these impurities for
- stationary phase: end-capped octad«yl.silyl silica gelfor demonstration of compliance. See also 5.10. Control of impun"ties
chromategraphy R (5 um); in subs/anus f()T pharmaceutical use) C, F) G.
~
Mobile phase: NH2 F
- mobile phase A: acetom'tn7e R, methanol R, 0.1 per cent VIV
solution of glacial acetic acid R (5:35:60 VIVIV);
N I
- mobile phase B: acetonitrile R, 0.1 per cent VIV solution of
oJ.-. N
~
glacial acetic acid R, merhanol R (5:15:80 VIVIV);
Thno MobUe phase A MobUe phase B

(min) (per cent VIV) (per cent VIJI) OH OH
0-' [00 0
5 - 20 100 ..... 49 0--->51 A. .4-amino-I-(5-deoxy-p-D-ribofuranosyl) -5-f1uoropyrimidin-
20 - 30 49 2( 1H)-one (5'-deoxy-5-f1uorocytidine),
"
Injecu"on 10.u. of the test solution and reference
Identification of impun'ties Use the chromatogram obtained
impurities A, Band D.
Relative retention With reference to capecitabine (retention
time = about 17 min): impurity A = about 0.18; B. 1-(5-deoXY-P-D-ribofuranosyl)-5-f1uoropyrimidine-2,4
impurity B =about 0.19; impurities D and E =about 0.95. (IH,3H)-dione (5'-deoxy-5-f1uorouridine),
System suilability Reference solution (c):
impurities A and B; minimum 2.0 between the peaks due
to impurity D and capecitabine.
Calculau"on of percentage amwus:
- for each impurity) use the concentration of capecitabine in
- c011"<C,wn factor: multiply the peak area of impurity B by
1.3.
Limits:
- impun·lies A) B: for each impurity) maximum 0.3 per centj
- sum of impurities D and E: maximum 0.2 per cent; C. 1-(2,3-di-o-acetyl-5-deoxy-ll-n-ribofuranosyl)-4-amino-5-
- unspecified impun'ties: for each impurity) maximum fluoropyrimidin-2( 1H)-one,
0.05 per cent;
Water (2.5.32)
Maximum 0.3 per cent"
Inject 1.0 mL of a 0.200 g1mL solution of the substance to
be examined in methanol R.
Maximum 0.1 per cent, determined on 1.0 g in a platinum
crucible.
ASSAY
D. (2RS)-2-methylbutyl [1-(5-deoxy-P-n-ribofuranosyl)-5-
fluoro-2-oxo-l )2-dihydropyrimidin-4-yl]carbamate,
Injecu·on Test solution and reference solution (a).
Calculate the percentage content of C15H22FN306 taking
into account the assigned content of caoecuabine CRS.
IMPURITIES
Specified impurities A, B D, E.
J
Otherdetectable impul1iies (thefollowing substances would, if

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2022 Caprylocaproyl Macrogolglycerides 1-423
CHARACTERS
Appearance
Pale-yellow, oily liquid.
Solubility
Dispersible in hot water, freely soluble in methylene chloride.
Density: about 1.0 at 20 "C.
Refractive index
About 1.4 at 20 "C.
IDENTIFICATION
E. 3-methylbutyl [1-(5-<1eoxy-Jl-D-ribofuranosyl)-5-fluoro-2- A. Thin-layer chromatography (2.2.27).
oxo-I,2-dihydropyrimidin-4-yl]carhamateJ Test solmion Dissolve l.0 g of the substance to be examined
in methylene chloride R and dilute to 20 mL with the same
solvent.
Place TLC silica gelplace R.
Mobile phase hexane R, ether R (30:70 VIV).
Application 50 ~L.
Drying In air.
Detection Spray with a 0.1 gIL solution of rhodamine B R in
ethanol (96 per un\! R and examine in ultraviolet light at
365 urn.
Results The chromatogram shows a spot due to triglycerides
with an R p value of about 0.9 (RJI 1) and spots due to
F. pentyl [5-fluoro-I-[(3aR,4R,6R,6aR)-6-methyl-2-
1,3-diglycerides (R" 0.7), to 1,2-diglycerides (R" 0.6), to
oxotetrahydrofuro [3,4-d] [1,3] dioxol-4-yl]-2-oxo-1 ,2-
monoglycerides (RJt 0.1) and [0 esters of macrogol (RJ , 0).
dihydropyrimidin-4-yl]carbamate,
B. Hydroxyl value (see Tests).
C. Saponification value (see Tests).
D. Composition of fatty acids (see Tests).
TESTS
Viscosity (2.2.9)
Carry out the determination at 20 ± 0.5 "C.
Ethylene oxide units Type of macrogo I Viscosity
per molecule (nominal (mPa-s)
value)
4 200 301050
6 300 60 to 80
8 400 80 to no
G. pentyl [1-(2,3-di-D-acetyl-5-deoxy-p-D-ribofuranosyl)-5-
fluoro-2-oxo-J ,2-dihydropyrimidin-4-yl]carbamate. Acid value (2.5.1)
____________________ Ph,,, Maximum 2.0, determined on 2.0 g.
Hydroxyl value (2.5.3, MethodA)
Use 1.0 g.
Caprylocaproyl *** Ethylene oxide units Type of macrogoJ Hydroxyl value

*** *** per molecule (nominal
Macrogolglycerides *** value)
4 200 80 to 120
6 300 14010 LBO
Action and use 8 400 170 to 205"
Excipient.
Ph,,, _ Peroxide value (2.5.5, Method A)
Maximum 6.0, determined on 2.0 g.
DEFINll10N Saponification value (2.5.6)
Mixtures of monoesters, diesters and triesters of glycerol and Use 2.0 g.
monoesters and diesters of macrogols with a mean relative
molecular mass between 200 and 400. Ethylene oxide units Type of macrogo I Saponification value
They are obtained by partial alcoholysis of medium-chain per molecule (nominal
value)
triglycerides using macrogol, or by esterification of glycerol
4 200 265 to 285
and macrogol with caprylic (octanoic) acid and capric
6 300 170 to L90
(decanoic) acid, or by mixingglycerol esters and condensates
of ethylene oxide with caprylic acid and capric acid. They 8 400 85 to 105
may contain free macrogols.
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1-424 Captopril 2022
Alkaline impurities
Introduce 5.0 g into a test-tube and carefully add a mixture, Captopril
neutralised if necessary with 0.01 M hydrochiotic acid or with (Ph. Eur. monograph 1079)
0.01 M sodium hydroxide, of 0.05 mL of a 0.4 gIL solution of
bromophenol blue R in ethanol (96 per cenO R, 0.3 mL of
water R and 10 mL of ethanol (96 per cent) R. Shake and
allow to stand. Not more than 1.0 mL of 0.01 M hydrochloric
acid is required to change me colour of the upperlayer to
yellow.
Free glycerol c.,H15NO, S 217.3 62571-86-2
Dissolve 1.20 g in 25.0 mL of methylene chloride R. Heat if Action and use
necessary. After cooling, add 100 mL of water R. Shakeand Angiotensin converting enzyme inhibitor.
add 25.0 mL of periodic acetic acidsolution R. Shake and allow Preparations
to stand for 30 min. Add 40 mL of a 75 gIL solution of Captopril Oral Solution
potassium iodide R. Allow to stand for 1 min. Add 1 mL of Captopril Tablets
stan:h solution R. Titrate the iodine with 0.1 M sodium
PhEIl ~
thiosulfate. Carry out a blanktitration.
1 mL of 0./1\1 sodium thiosulfate is equivalent [0 2.3 mg of DEFINITION
glycerol. (2S)-I- [(2S) -2-Methyl-3-sulfanylpropa noyl]pyrrolidine- 2-
Composition of fatty acids (2.4.22, Method A) carboxylic acid.
Composition of thefatty-acid fraction of the substance:
Content
- capl"Ok acid: maximum 2.0 per cent;
- caprylic acid: 50.0 per cent to 80.0 per cent;
- capm acid: 20.0 per cent to 50.0 per cent; CHARACTERS
- loutic add: maximum 3.0 per cent; Appearance
- myristic acid: maximum 1.0 per cent. White or almost white, crystalline powder.
Ethylene oxide and dloxan (2.4.25) Solubility
Maximum I ppm of ethylene oxide and maximum 10 ppm Soluble in water, freely soluble in methanol and in methylene
of dioxan. chloride. It dissolves in dilute solutions of alkali hydroxides.
Water (2.5.12) IDENfIFICATlON
Maximum 1.0 per cent, determined on 1.00 g. Use a mixture
of 30 volumes of anhydrous methanol Rand 70 volumes of
methylene chloride R as solvent B. Infrared absorption spectrophotometry (2.2.24).
Total ash (2.4.16) Comparison captopril CRS.
Maximum 0.1 per cent. TESTS
LABELLING Solution S
The labelstates the type of macrogol used (mean relative Dissolve 0.5 g in carbon dioxide-free water R and dilute to
molecular mass) or the numberof ethylene oxide units per 25 mL with the same solvent.
molecule (nominal value). Appearance of solution
FUNCTIONALITY-RELATED CHARACTERISTICS Solution S is clear (2.2.1) and colourless (2.2.2, Method II).
This seaion provides infonnation on characteristks that are pH (2.2.3)
recognised as being relevant amtrol parameters for one or more 2.0 to 2.6 for solution S.
functions of the substance when used as an excipient (see chapter Speclfle optical rotation (2.2.7)
5.15). Some of thecharaaerisucs described in 'he Functionality- -132 to -127 (dried substance).
pan of the monograph since they also represen: mandatory quality Dissolve 0.250 g in anhydrous ethanol R and dilute to
ctitena. In such cases, a cross-reference W the tests described in the
mandatory part is included in the Fun,tionah"ty-related hnpurlty F .
charaaenstics section. Control of the characteristics can wntn·bute Gas chromatography (2.2.28).
to thequality of a medicinal product by improving the consistency Reagent solution Add 2.8 mL of acetylchloride R dropwise to
of the mamifa<luring process and theperformance of the medicinal 17.2 mL of anhydrous methanol R at 0 ·C and mix. $low to
produc during use. Where control methods arecited, they are stand for 20 min at room temperature beforeuse. '
recognised as being suitable for the purpose, butother methods can Test solution Introduce 20.0 mg of the substance to be
also be used. Wherever results for a particular charaaeriuic are examined into a vial and add 1.0 mL of the reagent solution.
reported, the control method must be indicated. Mix and heat at 60 ·C for 30 min. Evaporate to dryness
Thefollowing characteristics may be relevant for caprylocaproyl under a stream of nitrogen R. Dissolve the residue in 0.5 mL
macrogolglycerides used as se/f-emulsijyJ"g agents and solubl7isers. of ethylacetate R, add 0.5 mL of pentaftuoropropionic
Hydroxyl value anhydride R, mix and heat at 60°C for 30 min. Evaporate to
(see Tests). dryness under a stream of nitrogen R. Dissolve the residue in
Saponification value 1.0 mL of butyl acetate R.
(see Tests). Reference solution (a) Dissolve the contents of a vial of
Composition of fatty acids captopril for system suitabl7ity CRS (containing impurity F) in
(see Tests). I mL of the reagent solution. Prepare as described for the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE,; test solution.
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2022 Captopril 1-425
Reference solution (b) Mix 0.25 mL of reference solution (a) and add 230 J1L of 0.05 iH iodine. If the solution is not
and 0.75 mL of butyl acetate R. colourless, add 0.1 M sodium thiosulfate dropwise until it
Column: becomes colourless, and dilute to 50 mL with the solvent
- material: fused silica; mixture. Dilute 5 mL of this solution to 20 mL with the
- size: / = 25 m, 0 ;;;; 0.32 mm; solvent mixture.
- stationary phase: phenyl(5)methyl(95)polysiloxane R (film Reference solution (d) Dilute 1.0 mL of the test solution to
thickness I I'm). 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
Carrier gas helium for chromatography R. solution to 10,0 mL with the solvent mixture.
FkYw rate 1.2 mUmin. Column:
Split ratio 1:20. - size: 1= 0.3 ID, 0 = 3,9 mm;
- stationary phase: irregular end-capped oaade<ylsilyl sihca gel
Temperature: for chromaUJgraphy R (10 urn);
Tim. Temperature
(min) ("Cl Mobile phase:
Colwnn 0-10 200
- mobile phase A: phosphoric acidR, waterfor
10· 14 200 ...... 240
chromaUJgraphy R (0.08: 100 VIII);
-=- mobile phaseB: phosphoric acidR) aceumim'/e R1, waterfor
14 - 34 240
Injection port 270
chromatography R (0.08:50:50 VIVIII);
Detector 300
(min) (per cent VI1') (per cent VII')
Detection Flame ionisation. 0-5 90 10
Inje<tion I I'L. 5·20 90 ...... 50 10 -;0 50
Relative retention With reference [Q captopril (retention 20 ~ 45 50 50
time = about 6 min): impurity F = about 0.96.
Systemsuitability: FkYw rate 1.5 mIJmin.
- resolution: minimum 1.5 between the peaks due to Detection Spectrophotometer at 210 om.
impurity F and capropril in the chromatogram obtained
Injection 25 ~L.
- signal-to-noise ratio: minimum 10 for the peak due [0 Identification of impuniies Use the chromatogram obtained
impurity F in the chromatogram obtained with reference
impurities B, C, D and J; use the chromatogram obtained
solution (b).
Calculate the percentage content of impurity F using the impurity Hj use the chromatogram obtained with reference
following expression: solution (e) to identify the peak due to impurity A.
A Relativeretention With reference to captopril (retention
x IOO time = about 15 min): impurity C = about 0.6j
A+B
impurity D = about 0.8j impurity E ;;;: about 0.9;
A area of the peak due [0 impurity F in the chromatogram
obtained with the test solution;
=
impurity B about 1.17; impurity J =about 1.22;
B area of the peak due to captopril in the chromatogram obtained impurity A = about 1.7.
with the test solution, System suitability:
Limit: impurities Band J in the chromatogram obtained with
- impun"ty F: maximwn 0.2 per cent. reference solution (a);
Related substances - resolution: minimum 2.0 between the peaks due to
Liquid chromatography (2.2.29). impurity E and captopril in the chromatogram obtained
Solvent mixture phosphoric acidR, acetonitrile Rl, waterR with reference solution (b),
(0.08:10:90 VIVIII). Limits:
Testsolution Dissolve 0.125 g of the substance to be - impun'ty A: not more than 10 times the area of the
examined in the solvent mixture and dilute to 25.0 mL with principal peak in the chromatogram obtained with
the solvent mixture, reference solution (d) (1.0 per cent);
- impun'ty J: not more than 2.5 times the area of the
Reference solution (a) Dissolve 4.0 mg of captopril
impun'ty J CRS, 5.0 mg of captopril impuniy B CRS, 5.0 mg
of capropril impurity C CRS and 5.0 mg of captopril
- impurities B, C, D: for each impurity, not more than
impurity D CRS in the solvent mixture and dilute to 50.0 mL
with the solvent mixture. Dilute 1.0 mL of the solution to
20.0 mL with the solvent mixture, Prepare immediately
(0.15 pec cent);
before use.
- impurity E: not more than 1.5 times the area of the
Reference solution (b) Dissolve 5 mg of the substance to be principal peak in the chromatogram obtained with
examined and 5 mg of captopril impurity E CRS in reference solution (d) (0.15 per cent);
acetonitrile R and dilute to 25 mL with the same solvent. - unspecified impuniies: for each impurity, not more than the
Dilute 4 mL of the solution to 50 mL with the solvent area of the principal peak in the chromatogram obtained
mixture. with reference solution (d) (0010 per cent);
Reference solution (c) In order to prepare impurity A in situ, - total: maximum 1.2 per cent;
introduce 1.0 mL of the test solution into a volumetric flask
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1-426 Captopril 2022
- disregard limit: 05 times the area of the principal peak in

the chromatogram obtained with reference solution (d)
(0.05 per cent).
Maximum 1.0 per cent, determined on 1.000 g by drying in G. (2RSj-3-(acetylsulfanyl}-2-methylpropanoic acid,
vacuo at 60 °C at a pressure not exceeding 0.1 kPa for 3 h.
o 0 0 H
Maximwn 0.2 per cent, determined on 1.0 g. H,cASySyN-\,Co,H
ASSAY
HCH3 HCH3 U
Dissolve 0.150 g in 30 mL ofwarer R. Titrate with 0.05 M
H. (2Sj-I-[(2Sj-3-[[(2R)-3-(acetylsulfanyl}-2-
iodine, determining the end-pointpotentiometrically (2.2.20).
methylpropanoyl]sulfanyl]-2-methylpropanoyl]pyrrolidine-
Use a combined platinum electrode.
2-carboxylic acid,
I mL of 0.05 M iodine is equivalent to 21.73 mg of
C.H 15NO,S.
IMPURITIES
Specified impurities A, H, C, D, E, F, J.
Other detectable impurities (thefol/Qwing substances would, if
presetu at a sufficient level, be delated by oneor other of the tests
I. (2Sj-I-[(2Sj-3-[[[(2Sj-I-[(2Sj-2-methyl-3-
in the manograph. They arelimiud by the general acceptance
sulfanylpropanoyl]pyrrolidin-2-yl]carbonyl]sulfanyl]-2-
cnienOn for otherlunspedfied l'mpuniies and/or by the general
methylpropanoyl]pyrrolidine-2-casboxylic acid,
manograph Substances for pharmaceutical use (2034). It is
therefore not necessary to identiJY these impurities for
in substances for pharmaceutical use) G, H, I, L, kf. N, O.
Ho,C · 1.X~
H
~Yo 1 OH..Co,H J.
O N~ '8-8 .... ~N
(2S)-I-[(2Sj-3-(acetylsulfanyl}-2-methylpropanoyl]
H" c~ H CH 3 pyrrolidine-2-casboxylic acid (acetylcaptopril),
A. 1,1'-[disulfanediylbis[(2Sj-2-methyl-I-oxopropane-3,1-
diyllJbis[(2S)-pyrrolidine-2-casboxylic] acid (captopnl
disulfide),
o L. 1,1'-[methylenebis[sulfanediyl[(2Sj-2-methyl-l-
~ Jl H Co,H
oxopropane-3, I-diylll]bis[(2Sj-pyrrolidine-2-casboxylic]
8,/ A ·w\;: acid,
H CH3U
B. (2Sj-I-[(2Sj-3-bromo-2-methylpropanoyl]pyrrolidine-2-
carboxylic acid,
~Co,H
HS /\ andenanliomer M. (2Sj-I-[(2Sj-3-[[(2Sj-2-casboxypropyl]disulfanYl]-2-
H CH3
methylpropanoyl]pyrrolidine-2-casboxylic acid,
C. (2RSj-2-methyl-3-sulfanylpropanoic acid,
~COzH
Br /\ and enanliomer
H CH3
D. (2RSj-3-bromo-2-methylpropanoic acid, N. 3,3'-disulfanediylbis[(2Sj-2-methylpropanoic] acid,
O. 1,1'-[propane-2,2-diylbis[suifanediyl[(2Sj-2-methyl-l-
E. (2Sj-I-(2-methylpropanoyl}pyrrolidine-2-casboxylic acid, oxopropane-3,I-diylJ]]bis[(2Sj-pyrrolidine-2-carboxylic]
acid.
~
HS' X
1-N-":H .Co,H _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw
H CH3 U
F. (2Sj -1- [(2R) -2-methyl-3-sulfanylpropanoyI]pyrrolidine-2-
carboxylic acid (epi-captopril),
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2022 Carbamazepine 1-427
Plate cellulose for chromatography R as the coatingsubstance.

Carbachol
Mobile phase water R. methanol R (10:90 VIV).
(Ph. Bur. monograph 1971) Application 10 ~L.
Deuelopment Over 2/3 of the plate.
Detection Spray with potassium iodobismulhate solution R3.
182.7 51-83-2
Limits In the chromatogram obtainedwith test solution (a):
Action and use - any impun'ty: any spot, apart from the principal spot, is
Cholinoceptor agonist. not more intense than one or other of the 2 principal
spots in the chromatogram obtained with reference
PhEu _ solution (b) (I per cent). Compare the spots with the spot
of the most appropriate colour in the chromatogram
DEFINITION
obtained with reference solution (b).
2-(Carbamoyloxy)-NJlV.N-trimethylethanaminium chloride.
Content
99.0 per cent to 101.5 per cent (dried substance). an oven at 105°C for 2 h.
CHARACTERS Sulfated ash (2.4.14)
Appearance Maximum 0.1 pee cent) determined on 1.0 g of the residue
White or almost white, crystalline, hygroscopic powder. obtained in the test for loss on drying.
Solubility ASSAY
Very soluble in water, sparingly soluble in alcohol) practically
Dissolve 0.150 g in a mixture of 10 mL of anhydrous acetic
insoluble in acetone.
acidRand 40 mL of acetic anhydride R. Titrate with 0.1 M
IDENTIFICATION perchloric add. Determine the end-pointpotentiometrically
First identification: A, C. (2.2.2(J).
Second identification: B, C. I mL of 0.1 M perchloric acid is equivalent to 18.27 mg of
A. Infrared absorption spectrophotometry (2.2.24). C6HlSClNzOZ'
Comparison carbachol CRS. STORAGE
B. Examine the chromatograms obtained in the test for In an airtight container, protected from light.
related substances. IMPURITIES
with test solution (b) is similar in position, colour and size to
C. 0.5 mL of solution S (see Tests) gives reaction (a) of A. 2-hydroxy-N,N,N-trimethylethanaminium chloride
chlorides (2.3.1). (choline chloride}.
TESTS _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>Eu
Solution S
Appearance of solution Carbamazepine ******
**
Acidity or alka1lnJty
(Ph. Bur. monograph 0543) *** *
To 2.0 mL of solution S, add 0.05 mL of methyl red mixed
solution R. Not more than 0.2 mL of 0.01 M hydrochloric acid v "I, 0
or 0.01 M sodium hydroxide is required to change the colour -<"
of the indicator. NANH'
Related substances "- ,f'~
Thin-layer chromatography (2.2.27). "0--
Prepare the soluNons immediately before use.

Testsolution (a) Dissolve 0.20 g of the substance to be 236.3 298-46-4
solvent. Action and use
Test solutioll (b) Dilute 2.0 mL of test solution (a) to Antiepileptic.
20.0 mL with methanol R. Preparations
Reference soluNon (a) Dissolve 20 mg of carbachol CRS in Carbamazepine Oral Suspension
methanol R and dilute (0 5.0 mL with the same solvent. Carbamazepine Tablets
Reference sduuon (b) Dissolve 8 mg of choline chloride Rand Carbamazepine Chewable Tablets
8 mg of acetylcholille chloride CRS in methanol R and dilute to Carbamazepine Prolonged-release Tablets
10.0 mL with the same solvent. Dilute 5.0 mL to 10.0 mL
with methanol R.
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1-428 Carbamazepine 2022
PhE~ _ Relative retention With reference to carbamazepine

DEFINITION (retention time = about 10 min): impurity A = about 0.9;
impurity E = about 3.5.
SH-Dibenzo[bJ)azepine-S-carboxamide.
System suitability:
Content
impurity A and carbamazepine in the chromatogram
CHARACTERS obtainedwith reference solution (a).
Appearance Limits:
Whiteor ahnost white, crystalline powder. - impurities A.. E: for each impurity, not more than
Solubility 1.5 times the area of the corresponding peak in the
Very slightly soluble in water, freely soluble in methylene chromatogram obtained withreference solution (a)
chloride, sparingly soluble in acetone and in ethanol (O.IS per cent);
(96 per cent). - unspecified impurities: not more than the area of the peak
It shows polymorphism (5.9). The acceptable crystalline form due to carbamazepine in the chromatogram obtained with
corresponds to carbamazepine CRS. reference solution (a) (0.10 per cent);
- wUll: not more than 5 times the area of the peak due to
IDENTIFICATION carbamazepine in the chromatogram obtained with
A. Melting point (2.2.14): 189"C to 193 "C. reference solution (a) (0.5 per cent);
B. fnfrared absorption spectrophotometry (2.2.24). - disregard limit: 0.5 times the area of the peak due to
Comparison carbamazepine GRS. carbamazepine in the chromatogram obtained with
Preparation Examine the substances as discs without prior reference solution (a) (0.05 per cent).
treatment. Chlorides (2.4.4)
Masimusn 140 ppm.
TESTS
Acidity or alkalinity Suspend 0.71S g in 20 mL of warer R and boil for 10 min.
To 1.0 g add 20 mL of carbon dioxide-free waterR, shake for Cool and dilute to 20 mL with water R. Filler through a
IS min and filter. To 10 mL of the filtrate add O.OS mL of membrane filter (nominal pore size 0.8 urn). Dilute 10 mL of
phenolphthalein solution RI and 0.5 mL of 0.01 M sodium the filtrate to 15 mL with water R.
hydroxide; the solution is red. Add 1.0 mL of 0.01 M Loss on drying (2.2.32)
hydrochloric acid; the solution is colourless. Add O.IS mL of Masimusn 0.5 per cent, determined on 1.000 g by drying in
methyl red solution Rj the solution is red. an oven at 105°C for 2 h.
Liquid chromatography (2.2.29). Maximum 0.1 per cent, determined on 1.0 g.
Test solution (a) Dissolve 60.0 mg of the substance to be ASSAY
examined in methanol R and dilute to 20.0 mL with the same Liquid chromatography (2.2.29) as described in the test for
solvent. Sonicate. Dilute 10.0 mL of this solution to 20.0 mL related substances with the following modifications.
with water R. Injection Test solution (b) and reference solution (b).
Test solution (b) Dilute 10.0 mL of test solution (a) 10 System suitability:
50.0 mL with a mixture of equal volumesof melhanol R and - repe.atabilit;y: reference solution (b).
water R.
Calculate the percentage content of C15H12N20 from the
Reference solution (a) Dissolve 7.5 mg of declared content of carbamazepim CRS.
carbamazepine CRS, 7.5 mg of carbamazepine impurity A CRS
and 7.5 mg of iminodibenzyl R (impurity E) in methanol R STORAGE
and dilute to 100.0 mL with the same solvent. Dilute 1.0 mL In an airtight container.
of this solution to 50.0 mL with a mixture of equal volumes IMPURITIES
of methanol R and water R. Specified impurities A, E.
Reference solution (b) Dissolve 60.0 mg of Other detectable impun'ties (thefollowing substances would, if
catbomasepine CRS in methanol R and dilute to 20.0 mL with present at a sufficient level, be deuaed by oneor other of the tests
the same solvent. Sonicate. Dilute 5.0 mL of this solutionto in the monograph. They are limited by the general acceptance
50.0 mL with a mixture of equal volumesof methanol R and ctitenon for otherlunspecified impW'liies and/or by thegeneral
water R. monograph Substances for pharmaceutical use (2014). It is
Column: therefore not necessary to identify these impurities for
- size: 1== 0.25 m, 0 = 4.6 mm; demonstration of compliance. See also 5.10. Control ofimpuniies
- stationary phase: cyanosilyl silica gelfor chromatography R in substances for pharmaceutical use) BJ C, D.. FJ G.
(10 urn),
Mobaephase tetrahydrofuran R, methanol RJ,.waterfor
chromatography R (3:12:8S VIVIV); to 1000 mL of this
solution add 0.2 mL of anhydrous formic acid Rand 0.5 mL
of triethylamine R.
Injection 20 ~ of test solution (a) and reference A. 10,II-dihydro-SH-dibenzo[bJ]azepine-S-carboxarnide
solution (a). (IO,II-dihydrocarbamazepine),
Run time 8 times the retention time of carbamazepine.
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2022 Carbasalate Calcium 1-429
"
%
Carbasalate Calcium
"N
H,C I : I
B. 9-melhylacridine,
458.4 5749-67-7
Action and use

Salicylate; non-selective cycle-oxygenase inhibitor;
antipyretic; analgesic; anti-inflammatory.
C. (5H-dibenzo[bJ)azepin-5-ylcarbonyl)urea (N- PIlE" _
carbamoylcarbamazepine),
DEFINmON
Equimolecular compound of calcium di[2-(acetyloxy)
benzoate] and urea.
Content
99.0 per cent [0 101.0 per cent (anhydrous substance).
CHARACTERS
Appearance
D. 5H-dibenzo[bJ)azepine (iminostilbene), White or almost white, crystalline powder.
Solublllty
Freelysoluble in water and in dimethylfonnamide, practically
insoluble in acetone and in anhydrous methanol.
ProCUI thesubstance from moisture during handling. Examination
in aqueous solutions has 10 beperfonned immediately after
preparation.
IDENl1FlCATION
E. 1O,11-dibydro-5H-dibenzo[bJ)azepine (iminodibenzyl),
Secondidentification: A, C, D, E.
(1.1.15).
Test solution Dissolve 0.250 g in water R and dilute to
100.0 mL with the same solvent. To 1.0 mL or the solution
add 75 mL of water Rand 5 mL of dilute hydrochloric acid R,
mix and dilute to 100.0 mL with water R. Examine
immediately.
F. 5H-dibenzo[bJ)azepine--5-carbonyl chloride Spectral range 220-350 nm.
(5-chlorocarbonyliminostilbene),
Absorption maxima At 228 nm and 276 nm.
Specific absorbance at the absorption maxima:
r "" 0 - at 228 nm: 363 to 379,
-'"
N
..Jl. NHz - at 276 nm: 49 to 53.
'"
Be
f'
~
j Comparison Ph. Eur. reference spectrum of carbasakJte calcium.
C. Dissolve 0.1 gin 10 mL of 'l(,'ater R, boil for 2 min and
cool. The solution gives reaction (a) ofsalicylates (2.3.1).
G. 10-bromo-5H-dibenzo[bJ]azepine-5-carboxamide (10- D. Heat 0.2 g with 0.2 g of sodium hydro.•ide R; a yellow or
bromocarbamazepine). yellowish-brown colour is produced and the vapour turns red
_ _ _ _~ PIIE"
litmus paper R blue.
E. It gives reaction (a) of calcium (1.3.1).
TESTS
The solution is not more opalescent than reference
suspension II (1.1.1) and is colourless (1.1.1, Method II).
Dissolve 2.5 g in 50 mL of water R.
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1-430 Carbasalate Calcium 2022
Related substances Test solution Dissolve 1.0 g in 500.0 mL of water R.

liquid chromatography (2.2.29). Prepare the solutions Water (2.5.12)
immediately before use. Maximum 0.1 per cent, determined on 1.000 g. Use a
Solvent mixture phosphoric acid R, methanol R, acetonitrile for mixture of 15 mL of anhydrous methanol Rand 15 mL of
chromatography R (0.5:8:92 VIV/Y). dimelhylfonnamide R as the solvent.
Test solution Dissolve 0.100 g of the substance to be ASSAY
examined in 5 mL of the solvent mixture, sonicate for In a flask with a ground-glass stopper, dissolve 0.400 g in
15 min and dilute to 10.0 mL with the solvent mixture. 25 mL of water R. Add 25.0 mL of 0.1 M sodium hydroxide.
Filterthe solution through a membrane filter (nominal pore Close the flask and allow to stand for 2 h. Titrate with 0.1 M
size 0.45 urn). hydrochloric acid, using 0.2 mL of phenolphthalein solurion R.
Reference solution (a) Dissolve 10.0 mg of salicylic acidCRS Carry out a blank. titration.
(impurity C) in the solvent mixtuse and dilute to 100.0 mL I mL of 0.1 M sodium hydroxide is equivalent to 22.92 mg of
C19HISCaN209'
STORAGE
Reference solution (c) Dissolve 2 mg of carbasalate
impuniy B CRS in 20.0 mL of the solvent mixtuse. IMPURITIES
Reference solution (d) Dilute 1.0 mL of the test solution to Specified impurities B, C.
10.0 mL with the solvent mixtuse. Mix 1.0 mL of this Otherdetectable impurities (the following subS/anas would, if
solution with 5.0 mL of reference solution (a), add 1.0 rnL present at a sufficient level, be detected by oneor other of the tests
of reference solution (c) and dilute to 10.0 mL with the in the monograph. They all! limited by thegeneral acceptance
solvent mixture. criterion for other/unspecified impurities and/or by thegeneral
Column: monograph Substanas for pharmaceutical use (2034). It is
- size: 1= 0.25 m, 0 = 4.0 mmj therefore no: neussary to identify these impurities for
- stationary phase: spherical end-capped octaduy/silyl sUica gel demonstration of compliance. See also 5.10. Control of impuniies
for chromatography R (5 ~m); in substances for pharmaceutical use) A J D.
MobUe phase phosphoric acid R, aatonitri/e for
chromatography R, water R (0.5:40:60 V/V/Y).
FIcJw rate 1.8 mUmin.
solutions (b) and (d). A. 2-(acetyloxy)benzoic anhydride,
Run lime 8 times the retention time of acetylsalicylic acid.
Idendfiauion of impurities Use the chromatogram obtained
with reference solution (d) to identify the peaks due to
impurities Band C.
Relatwe retention With reference to acetylsalicylic acid
(retention time :;;; about 2 min): impurity C = about 1.3;
=
System suicability Reference solution (d):
- resolution: minimum 5.0 between the peaks due to B. 2-[[2-(acetyloxy)benzoyl]oxy)benzoic acid
acetylsalicylic acid and impurity C. (acetylsalicylsalicylic acid),
Limits:
I
OC
CO,H
- impurity C: not more than 5 times thearea of the
corresponding peak in the chromatogram obtained with "- OH
- impun·,y B: not more than 1.5 times the area of the
C. 2-hydroxybenzenecasboxylic acid (salicylic acid),
- unspecified impurilks: for each impurity, not more than
(0.05 per cent);
- rotal: not more than 7 times the area of theprincipal peak
to the chromatogram obtained with reference solution (b)
(0.7 per cent);
- disregard limit: 0.3 times the area of the principal peak in D. 2-[(2-hydroxybenzoyl)oxy]benzoic acid (salicylsalicylic
the chromatogram obtained with reference solution (b) acid).
(0.03 per cent). ________ ~ ""E"
Sodium
Atomic emission spectrometry (2.2.22, Method 1).
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2022 Carbidopa 1-431
Using an ultrasonic bath} dissolve completely 0.250 gin

Carbidopa aluminium chloride solution R and dilute to 25.0 mL with the
(Ph. Ellr. monograph 0755) same solution.
Hydrazlne
• H,O Test solution (a) Dissolve 0.50 g in dilllte hydrochiotic acidR
and dilute to 2.0 mL with the same acid.
Test sollltion (b) Place 25 g of strongly basic anion-exchange
resin R into each of 2 conical flasks with ground-glass
244.2 38821-49-7 stoppers. To each} add 150 mL of carbon dioxide-free water R
and shake from time to time during 30 min. Decant the
Action and use liquid from both flasks and repeat the process with further
Dopa decarboxylase inhibitor. quantities} each of 150 ml., of carbon dioxide-free water R.
Preparation Take two 100 rnL measuring cylinders 3.5-4.5 em in internal
Co-careldopa Tablets diameter and label these A and B. Into cylinder A} transfer as
PhE" _
completely as possible the resin from 1 conical flask using
60 mL of carbon dioxide-free waterR; into cylinder B, transfer
DEFINITION the 2nd quantity of resin, this time using 20 mL of carbon
(2S)-3-(3,4-Dihydroxyphenyl)-2-hydrazino-2- dioxide-free water R.
methylpropanoic acid monohydrate. Into each cylinder, insert a gas-inlet tube, the end of which
Content has an internal diameter of 2-3 mm and which reaches
98.5 per cent to 101.0 per cent (dried substance). almost to the bottomof the cylinder. Pass a rapid stream of
nitrogen for chromawgraphy R through each mixture so that
CHARACTERS homogeneous suspensions are formed, After 30 min, without
Appearance interrupting the gas flow, add 1.0 mL of test solution (a) to
White or yellowish-white powder. cylinder Ai after 1 min stop the gas flow lnro cylinder A and
Solubility transfer the contents, through a moistened filter paper, into
Slightly soluble in water, very slightly soluble in ethanol cylinder B. After 1 min, stop the gas flow to cylinder Band
(96 per cent), practically insoluble in methylene chloride. pour the solution irrunediateJy through a moistened filter
It dissolves in dilute solutions of mineral acids. paper into a freshly prepared mixture of I mL of a 200 WL
solution of salicylaldehyde Rin methanol Rand 20 mL of
IDENTIFICATION
phosphate bllffer soilltion pH 5.5 R in a conicaillask; shake
First identification: A. C.
thorougWy for 1 min andheat in a water-bath at 60 DC for
Second identification: A} B} D, E. 15 min. The liquidbecomes clear. Allow to cool, add
A. Specificoptical rotation (see Tests). 2.0 mL of toluene R and shake vigorously for 2 min. Transfer
B. Ultraviolet and visible absorption spectrophotometry the mixture into a centrifuge tube and centrifuge.
(2.2.25). Separate the toluene layer in a 100 mL separating funnel and
Test solution Dissolve 50 mg in an 8.5 gIL solution of shake vigorously with 2 quantities, each of 20 mL, of a
hydrochloric add R in me/hanoi R and dilute to 100.0 mL with 200 gIL solution of sodium metabisll/fite R and finally with
the same solution. Dilute 10.0 mL of this solution to 2 quantities, each of 50 ml., of water R. Separate the toluene
100.0 mL with an 8.5 gIL solution of hydrochloric add R in layer.
methanol R. Reference sduuon (a) Dissolve 10 mg of hydrozine sulfate R
Spectral range 230-350 run. in dilllte hydrochloric add R and dilute to 50 mL with the
Absorption maximum At 283 nm. same acid. Dilute 1.0 mL of this solution to 10.0 mL with
diluu hydrochlom add R.
Specific absorbance at the absorption maximum 135 to 150
(dried substance). Reference solution (b) Prepare the solution at the same time
and in the same manner as described for test solution (b)
using 1.0 mL of reference solution (a) instead of 1.0 mL of
Comparison carbidopa CRS. test solution (a).
D. Shake vigorously about 5 mg with 10 mL of water R for Plate TLC silanised silica gelplate R.
I min and add 0.3 mL of feme chloride sohuion R2.
Mobile phase waterR, methanol R (10:20 VfII).
An intense green colour is produced, which quicldy changes
to reddish-brown. Application 10 J.lL of test solution (b) and reference
solution (b).
E. Suspend about 20 mg in 5 mL of water R and add 5 mL
of cupri-tartasic solution R. On heating} the colour of the Development Over 1/2 of the plate.
solution changes [0 dark brown and a red precipitate is Drying In air.
formed. Detection Examine in ultraviolet lightat 365 nm.
TESTS Limit:
Appearance of solution - hydrazine: any spot showing a yellowfluorescence is not
The solution is clear (2.2.1) and not more intensely coloured more intense than the corresponding spot in the
than reference solutionBY6 or B6 (2.2.2, il'letlwd If). chromatogram obtained with reference solution (b)
(20 ppm).
Dissolve 0.25 g in 25 mL of J M hydrochloric acid.
-26.5 to -22.5 (dried substance).
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1-432 Carbidopa 2022
Related substances - reporting threshold: 0.05 per cent.

Liquid chromatography (2.2.29). Prepare the solutions Loss on drying (2.2.32)
immediately before use. 6.9 per cent to 7.9 per cent, determined on 1.000 g by
Buffer solution Dissolve 6.9 g of sodium dihydrogen phosphate drying in an oven at 105 DC.
monohydrate R in 900 mL of waterfor chromarography R, Sulfated ash (2.4.14)
adjust to pH 2.2 with phosphoric acid R and dilute to Maximum 0.1 per cent, determined on 1.0 g.
1000 mL with waterfor chromarography R.
ASSAY
examined in the mobile phase, add 20 ~L of hydrochloric Dissolve 0.150 g with gentle heating in 75 mL of anhydrous
add R and dilute to 10.0 mL with the mobile phase. acetic add R. Titrate with 0.1 M perchloric acid, determining
the end-point potentiometrically (2.2.2/J).
100.0 mL with the mobile phase. Dilute 1.0 mL of this 1 mL of 0.1 M perchlonc acid is equivalent to 22.62 mg
solution to 10.0 mL with the mobile phase. of C lOHI4N,O,.
Reference ,olution (b) Dissolve 4 mg of carbidopa for system STORAGE
n
suitabi1liy CRS (containing impurities A, D, E, I and in the Protected from light.
mobile phase, add 4 ~L of hydrochloric acid R and dilute to IMPURITIES
2.0 mL with the mobile phase. specified ;mpuniies A, D, E, F, H, I, J.
Reference solution (c) , Dissolve the contents of a vialof Other detectable impurities (thefollowing substances would, if
carbiJopa impurity mixture CRS .(impurities F and H) in present at a sufficient level, be detected by oneor other of the tests
1.0 mL of reference solution (b). in the monograph. They are limited by thegeneral acceptance
Column: criterion for otherlunspedfied impurities and/or by the general
- size: 1= 0.15 m, 0 = 4.6 rom; monograph Substances for pharmaceutical use (2034). It is
- stationary phase: end-capped ethy/ene-bridged oaadecy/si/y/ therefore not necessary to idenufy these impurities for
silica gelfor chromarography (hybrid material) R (3.5 um), demonstration of compliance. See also 5.10. Conrrol of impun·ties
- temperature: 25°C. in substances for pharmaceutical use) B, C, G.
Mobile phase ethanol (96 perunl) R, buffer solution
~
(7:93 V/lo'). Co,H

HO
H,N \'H,
""- I
Deuction Spectrophotometer at 280 om.
OH
Injection 20~.
Run time 6 times the retention time of carbidopa. A. (2S)-2-amino-3-(3,4-dihydroxyphenyl)-2-methylpropanoic
Identification of impurities Use the chromatogram obtained acid (L-methyldopa),
impurities A, D + E, F, H, I and J. o
~
Relali've retention With reference to carbidopa (retention
.. OCH,
time =about 3 min): impurity A =about 0.9; impurities D ""- I H,N 'CH,
and E = about 1.9; impurity H = about 2.5; HO
impurity I = about 3.7; impurity J = about 4.0; OH
impurity F = about 4.4.
System suitability: B. methyl (2S)-2-amino-3-(3,4-dihydroxyphenyl)-2-
- resolution: minimum 1.5 between the peaks due to methylpropanoate,
impurity A and carbidopa in the chromatogram obtained
with reference solution (b); minimum 1.5 between the
peaks due to impurities I and J in the chromatogram
- signal-to-noise ratio: minimwn 30 for the principal peakin
Calculation of percentage contents: C. (2S)-2-hydsazino-3-(4-hydsoxy-3-methoxyphenyl)-2-
- cotreaion [actors: multiply the peak areas of the following methylpropanoic acid (3-D-methylcarbidopa),
impurities by the corresponding correction factor;
impurities D and E = 1.5; impurity I = 1.5; . o
=
impurity J 1.5;
- for each impurity, use the concentration of carbidopa in '" I OCH,
~ HN CH3
Limits:
HO OHd
- impurity J: maximum 0.25 per cent;
- sum of impuritks D and E: maximum 0.2 per cent;
- impuruies F, H, I: for each impurity, maximum D. methyl (2S)-2-(2-cyclohexylidenehydsazino)-3-(3,4-
0.15 per cent; dihydroxyphenyl)-2-methylpropanoate,
0.10 per cent;
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2022 Carbimazole 1-433
o PhE" ~ _
'" DEFINITION
Ethyl 3-methyl-2-thioxo-2,3-dihydro-1 H-imidazo le-I-
HO '"
OH carboxylate.
Content
E. methyl (2S)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2- 98.0 per cent to 102.0 per cent (dried substance).
methylpropanoate, CHARACTERS
o Appearance
White or yellowish-white, crystalline powder.
'" \
HN CH3
O »<: CH3
SolublIlty
Slightlysoluble in water, soluble in acetone and in ethanol
HO '" NHz
OH (96 per cent).
IDENTIFICATION
F. ethyl (2S)-3-(3,4-dihydroxyphenyl)-2-hydrazino-2- Firse identification: B.
methylpropanoate,
("("YCH, A. Melting point (2.2.14): 122 °C to 125 °C.
HO~ 0 Comparison carbimazok CRS.
OH
Co Thin-layer chromatography (2.2.27).
G.I-(3,4-dihydroxyphenyl)propan-2-one, Testsolution Dissolve 10 mg of the substance to be
examined in methylene chlOJ'ide R and dilute to 10.0 mL with
the same solvent.
Reference solution Dissolve 10 mg of carbimazole CRS in
H,CO methylene chloride R and dilute to 10.0 mL with the same
OH solvent.
Pial< TLC silica gel F 254 pIal< R.
H. (2S)-2-hydrazino-3-(3-hydroxy-4-methoxyphenyl)-2-
Mobilephase ace"'ne R, methylene chloride R (20:80 VIV).
methylpropanoic acid,
Application 10 fIL.
B<~Co,H Development Over 3/4 of the plate.
I "p HN \:H, Drying In air for 30 min.
HO \N~
Deuaion Examine in ultraviolet light at 254 nm.
OH
I. (2S)-3-(3-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2- with the test solution is similar in position and size to the
methylpropanoic acid, principal spot in me chromatogram obtained with the
reference solution.
B< D. Dissolve about 10 mg in a mixture of 0.05 mL of dilul<
~
Co,H hydrochloric acid Rand 50 mL of water R. Add 1 mL of
I ..# HN \:H 3
potassium iodobismuthate solution R. A red precipitate is
HO NH:2 formed.
OH TESTS
Related substances
J. (2S)-3-(2-bromo-4,5-dihydroxyphenyl)-2-hydrazino-2-
melhylpropanoic acid.
immediately before we.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Solvent mixture acetonitrile R, water R (20:80 VIV).
Carbimazole *** the solvent mixture.
*** *** Reference solution (a) Dissolve 5 mg of .hiamazol, CRS
(Ph. Bur, monograph 0884) *** (impurity A) in the solvent mixture and dilute to 100.0 mL
wah the solvent mixture. Mix 1 mL of the solutionwith
2 mL of the test solution and dilute to 10.0 mL with the
solvent mixture.
Reference solution (b) Dissolve 5.0 mg of thiamazol, CRS
(impurity A) in the solvent mixture and dilute to 100.0 mL
with the solvent mixture. Dilute 1.0 mL of the solution to
186.2 22232-5hS
Action and use Reference solution (c) Dilute 1.0 mL of the test solution to
Thionamide antithyroid drug. 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
Preparation solution to 10.0 mL with the solvent mixture.
Carbimazole Tablets
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1-434 Carbocisteine 2022
Reference solution (d) Dissolve 25.0 mg of carbimazole CRS

Carbocisteine ***
in the solvent mixtureand dilute to 50.0 mL with the solvent *** ***
mixture. (ph. Eur. monograph 0885) ***
Column:
- size: 1 = 0.15 m, 0 = 3.9 mm;
- Slalianary phase: end-capped OCladecylsilyl silica gelfor
chromalography R (5 pm).
Mobile phase acetonitrile R, waterR (10:90 VIV).
179.2 638-23-3
Flow rate 1 mlJrnin.
Detection Spectrophotometer at 254 run. Action and use
Inj«u"on 10 J.lL of the test solution and reference Mucolytic.
solutions (a), (b) and (c). PhE<r _
Run time J.5 times the retention time of carbimazole.
Identification of impurities Use the chromatogram obtained DEFINITION
with reference solution (a) to identify the peak due to Carbocisteine contains not less than 98.5 per cent and not
impurity A. more than the equivalentof 101.0 per cent of (2R)-2-amino-
3-[(carboxymethyl)sulfanyl]propanoic acid, calculated with
Relativeretention With reference to carbimazole (retention
reference to the dried substance.
= =
time about 6 min): impurity A about 0.2.
System suitability Reference solution (a): CHJ\RI\CTERS
- resolution: minimum 5.0 between the peaks due to A white or almost white, crystalline powder, practically
impurity A and carbimazole. insoluble in water and in alcohol. It dissolves in dilute
Limits: mineral acids and in dilute solutions of alkali hydroxides.
- ;mpun"ty A: not more than 2.5 times the area of the IDENTIFICATION
principal peak in the chromatogram obtained with Firse idenoficoiion: A, B.
reference solution (b) (0.5 per cent); Second identification: A, C, D.
with reference solution (c) (0.10 per cent); B. Examine by infrared absorption spectrophotometry
- total: maximum 0.6 per cent; (2.2.24), comparing with the spectrum obtained wiih
- disregard limit: 0.5 times the area of the principal peak in carbocisteine CRS. Examine the substances prepared as discs.
the chromatogram obtained with reference solution (c) C. Examine the chromatograms obtained in the test for
(0.05 per cent). ninhydrin-positive substances. The principal spot in the
chromatogram obtained with test solution (b) is similarin
Maximwn 0.5 per cent, determined on 1.000 g by drying in position, colour and size to the principal spot in the
a desiccator at a pressure not exceeding 0.7 kPa for 24 h. chromatogram obtained with reference solution (a).
D. Dissolve 0.1 gin 4.5 mL of dilute sodium hydroxide
solution R. Heat on a water-bath for 10 min. Cool and add
I mL of a 25 gIL solution of sodium nitroprusside R. A dark
ASSAY red colour is produced, which changes to brown and then to
liquid chromatography (2.2.29) as described in the test for yellow within a few minutes.
related substanceswith the following modification.
TESTS
Injection Test solution and referencesolution (d). Solution S
Calculate the percentage content of C1H1oN'202S taking into Disperse 5.00 g in 20 mL of waterR and add dropwise with
account the assigned content of carbimazo/e CRS. shaking 2.5 mL of sl>"Ollg sodium hydroxide solution R. Adjust
IMPURITIES to pH 6.3 with 1 M sodium hydroxide and dilute to 50.0 mL
Specified impurities A. with water R.
Solution S is clear (2.2.1) and colourless (2.2.2, Method Ii).
pH (2.2.3)
Shake 0.2 g with 20 mL of carbon dioxide-free waterR.
The pH of the suspension is 2.8 to 3.0.
A. l-methyl-IH-imidazole-2-thiol (thiamazole). Specific optical rotation (2.2.7)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r -32.5 to -35.5, determined on solution S and calculated
with reference to the dried substance.
suitable silica gel as the coating substance.
examined in d,1ute ammonia R2 and dilute to 10 mL with the
same solvent.
with water R.
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2022 Carbomers 1-435
Reference solution (a) Dissolve 10 mg of carbociueine CRS in CHARACTERS

dilute ammonia R2 and dilute to 50 mL with the same Appearance
solvent. White or almost white) fluffy, hygroscopic powder.
Reference solution (b) Dilute 5 mL of test solution (b) to Solubillty
20 mL with water R. Swells in water and in other polar solvents after dispersion
Reference solution (c) Dissolve 10 mg of corbociueine CRS and neutralisation with sodium hydroxide solution.
and 10 mg of arginine hydrochloride CRS in 5 mL of dilute IDENTIFICATION
ammonia R2 and dilute to 25 mL with water R. First idencfication: A.
Apply separately to the plate 5,~L of each solution. Allow the Secondidenufication: B, C, D.
plate to dry in air. Develop over a path of 15 em using a A. Infrared absorption spectrophotometry (2.2.24).
mixture of 20 volumes of glacial acetic acid R, 20 volumes of Main bands At 1710 ± 5 cm'", 1454 ± 5 cm',
waterRand 60 volumes of butanol R. Dry the plate in a 1414 ± 5 cm'", 1245 ± 5 em", 1172 ± 5 cm'",
current of warm all'. Spray with ninhydn"n solution R and heat
1115 ± 5 cm' and 801 ± 5 cm', with the strongest band
at 100 °C to 105 °C for 15 min. Any spot in the at 1710 ± 5 em-I.
chromatogram obtained with test solution (a), apart from the
B. Adjust a 10 gIL dispersion to about pH 7.5 with
principal spot, is not more intense than the spot in the
1 1M sodium hydroxide. A higWy viscous gel is formed.
(0.5 per cent). The test is not valid unless the chromatogram C. Add 2 mL of a 100 gIL solution of calcium chloride R,
obtained with reference solution (c) shows two clearly with continuous stirring, to 10 mL of the gel from
separated principal spots. identification test B. A white precipitate is immediately
produced.
Chlorides (2.4.4)
D. Add 0.5 mL of thymolbluesolution R to 10 mL of a
Dissolve 33 mg in 5 mL of dilute n;m'c acidR and dilute to
10 gIL dispersion. An orange colour is produced.
15 mL with water R. The solution, without further addition
Add 0.5 mL of cresol red solution R to 10 mL of a 10 gIL
of nitric acid, complies with me limit test for chlorides
dispersion. A yellow colour is produced.
(0.15 per cent).
TESTS
Sulfates (2.4.11)
Free acrylic acid
Dissolve 0.5 g in 5 mL of dilute hydrochloric acidR and dilute
to 15 mL with distilled water R. The solution complies with
the limit test for sulfates (300 ppm). Testsolution Mix 0.125 g of the substance to be examined
with a 25 gIL solution of aluminium potassium sulfate R and
Loss on drying (2.2.32) dilute to 25.0 mL with me same solution. Heat the
Not more than 0.5 per cent, determined on 1.000 g by suspension at 50 DC for 20 min with shaking) then shake the
drying in an oven at 105°C for 2 h. suspension at room temperature for 60 min. Cennifuge and
Sulfated ash (2.4.14) use the clear supernatant solution as the test solution.
Not more than 0.3 per cent, determined on 1.0 g. Reference solution Dissolve 62.5 mg of acrylic acid R in a
ASSAY 25 gIL solution of aluminium potassium sulfate R and dilute to
Dissolve 0.150 gin 10 mL of anhydrous formic acidR with 100.0 mL with the same solution. Dilute 1.0 mL of this
slight heating and shake until dissolution is complete. solution to 50.0 mL with a 25 gIL solution of aluminium
Add 50 mL of anhydrous acetic acid R. Titrate with 0.1 M porassium sulfate R.
percbloric acid, determining the end-point potentiometrically Column:
(2.2.20). - size: 1;;;; 0.12 m, 0 ;;;; 4.6 nun;
I mL of 0.1 M perchloric acid is equivalent to 17.92 mg of - stationary phase: ""tadecylsi!YI silica gelfor chromarography R
C,H.NO.S. (5 urn).
Mobile phase:
STORAGE
-c mobile phaseA: 1.361 gIL solution of potassium dihydrogen
Store protected from light.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
phosphate R, adjusted to pH 2.5 using dilute phosphoric
acidR;
- mobik phaseB: mixture of equal volumes of a 1.361 gIL
solution of potassium d;hydrog~n phosphate Rand acetonitrile
for chromarography R;
Carbomers
Time Mobile phase A Moblle phase B
(Ph. Eur. monograph 1299) (min) (per cent I'm (per cent I'm
Action and use O-S 100 0
Stabilizer in pharmaceutical products. 8-' 100 ..... 0 0 ..... 100
9 - 20 0 100
Preparation
Carbomer Eye Drops
Flow rate 1 mUmin.
PhE" _
DEFINITION Injection 20~.
High-molecular-mass polymers of acrylic acid cross-linked Retention lime Acrylic acid e about 6.0 min.
with alkenyl ethers of sugars or polyalcohols. Limit:
Content - acrylu; acid: not more than the area of the corresponding
56.0 per cent [0 68.0 per cent of carboxylic acid (-COzHJ peak in the chromatogram obtained with the reference
groups (dried substance). solution (0.25 per cent).
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1-436 Carbon Dioxide 2022
Benzene alsobe used. iWherever results for a particular characteristic are

Gas chromatography (2.4.24, System A). reponed, the control method must be indicated.
Solution A Dissolve 0.100 g of benzene R in dimethyl Thefollowing characteristics may be relevant for carbomers used as
sulfoxide R and dilute to 100.0 mL with the same solvent. viscosity-increasing agents and gelling agents.
Dilute 1.0 mL of the solution to 100.0 mL with water R. Apparent viscosity (2.2.10)
Dilute 1.0 mL of this solution 10 100.0 mL with water R. The nominal apparent viscosity is typically between
Test solution Weigh 50.0 mg of me substance to be 300 mf'a-s and 115 000 ml'a-s. For a product with a
examined into an injection vial and add 5.0 mL of water R nominal apparent viscosity of 20 000 ml'a-s or greater, me
and 1.0 mL of dimethyl sulfoxide R. apparent viscosity is typically 70.0 percent to 130.0 per cent
Reference solution Weigh 50.0 mg of the substance to be of me nominal value; for a productwith a nominal apparent
examined into an injection vial and add 4.0 mL of water R) viscosity of less than 20 000 ml'a-s, the apparent viscosity is
1.0 mL of dimethyl Slufoxide Rand 1.0 mL of solution A. rypically 50.0 per cent to 150.0 per cent of the nominal
Close the vials with a tight rubber membrane scqpper coated with value.
po/yutrafluoroelhylene and secure with an aluminium cn"mped Dry the substance to be examined in vacuo at 80 °C for I h.
cap. Shake to obtain a homogeneous dispersion. CarefuUy add 2.50 g of the previously dried substance 10 be
Static head-space conditions that may be used: examined 10 500 mL of water R in a 1000 mL beaker while
- equilibration temperature: 80°C; stirring continuously at 1000 ± 50 rlmin, with the stirrer
- equilibration lime: 60 nun, shaft set at an angle of 60° to one side of the beaker. Add the
- transfer-line temperature: 90°C. previously dried substance overa period of 45-90 s, at a
uniform rate,ensuring thatloose agglomerates of powder are
InjecuOn 1 mL of me gaseous phase of the test solution and
I mL of me gaseous phase of the reference solution; repeat
broken up, and continue stirring at 1000 ± 50 r/min fur
15 min. Remove the stirrer and place the beaker containing
these injections twicemore.
the dispersion in a water-bath at 25 ± I °C for 30 min.
System suitability: Insert the stirrer to a depth necessary to ensure thatair is not
- repeatabiJiIJ1: maximum relative standard deviation of the drawn into the dispersion and, while stirring at
differences in area between the analyte peaks obtained 300 ± 25 r/min, adjust to pH 7.3-7.8 by adding a 180 gIL
from the 3 replicate pair injections of the reference solution of sodium hydroxide R below the surface. The total
solution and the test solution is 15 per cent. volume of the 180 gIL solution of sodium hydroxide R used is
Limit: abour 6.2 mL. AUow2-3 min before the final pH
- benzene: the mean area of the peakdue to benzenein the determination. If the final pH exceeds 7.8, discard the
chromatograms obtained with the test solution is not preparation and prepare another using a smaller amount of
greater than 0.5 times the mean area of the peak due to sodium hydroxide. Return the neutralised preparation to the
benzene in the chromatograms obtained with the water-bath at 25°C for I h, then perform the viscosity
reference solution (2 ppm). determination without delay to avoid slight viscosity changes
Loss on drying (2.2.32) that occur75 min after neutralisation. Determine the
Maximum 3.0 per cent, determined on 1.000 g by drying in viscosity using a rotating viscometer with a spindle rotating at
vacuo at 80 °C for 60 min. 20 r/min, usinga spindle suitable for the expected apparent
viscosity.
Maximum 4.0 per cent, determined on 1.0 g. Carboxylic acid groups
See Assay.
ASSAY ___ ~ I'I>E"
Slowly add 50 mL of water R to 0.120 g whilst stirring and
heating at 60·C for 15 min. Stop heating, add 150 mL of
water R and continue stirring for 30 min. Add 2 g of
potassium chloride R and titrate with 0.2 M sodium hydroxide,
detennining the end-point potentiometrically (2.2.20). Carbon Dioxide
carboxylic acid (-C02H) groups.
Carbon Dioxide should bekept in approved metalcylinders which
STORAGE
arepaintedgrey and cany a labd stating 'Carbon Dioxide'.
In an airtight container. In addition, 'Carbon Dioxide' or the symbol 'C02' should be
FUNCTIONALITY-RELATED CHARACTERISTICS stencilled in paint on theshoulder of the cylinder.
This section provides information MI characteristics that are CO 2 44.01 124-38-9
recognised as being relevant control paramecm for one or more POE" _
functions of thesubstance when used as an excipient- (see chapter
5.15). Some of the charactetisda described in the Functionality- DEFINITION
related characteristics section may alsobepresent in the mandatory Content
partof the monograph since they also represent mandatory quality Minimum 99.5 per cent VIV of CO2 in the gaseous phase.
enteric. In such cases, a cross-reference to the tests described in the This monograph applies to carbon dioxide for medicinal use.
mandatory pan is included in theFunctionality-related
characteristics section. Control of the characteristics can contribute CHARACTERS
to thequality of a medicinal product by improving the consisceney Appearance
of the maU14a<turing prams and theperfo,",ame of the medicinal Colourless gas.
product dun·ng use. Where centrol methods are cited} they at\1 Solubility
recognised as being suitable for thepurpose, but other methods can At 20 °C and at a pressure of 101 kPa, 1 volume dissolves in
about I volume of water.
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2022 Carbon Dioxide 1-437
. - - - Chopper
Sample in Sample out
t
I I
D~
)-
( D=E )-
>
i
UV Source
tmertnm
Collimator
I I 0( Filler 350 nm
t
Photomultiplier
t
Amplifier
Figure 0375.-1.- UV Fluorescence AnalYser
PRODUCTION Reference gas (b) A mixture containing 2 ppm VIV of

Examine the gaseous phase. nitrogen monoxide R in carbon dioxide R 1 or in nitrogen RI.
If the test is performed an a cylinder af gas, keep the cylinder af Calibrate the apparatus and set the sensitivity using reference
the substance to be examinedat room temperature for not less than gases (a) and (b). Measure the content of nitrogen monoxide
6 h before canying out the tests. Keep the cylinder in the vertical and nitrogen dioxide in the gas to be examined.
position with the outlet valve uppermost. If nitrogen is used instead of carbon dioxide in reference
Carbon monoxide gas (b), multiply the result obtained by the quenching
Gas chromatography (2.2.28). correction factor in order to correct the quenching effect on
Gas to be examined The substance to be examined. the analyser response caused by the carbon dioxide matrix
effect
Reference gas A mixture containing 5 ppm VIV of carbon
monoxide R in nitrogen RI. The quenching correction factor is determined by applying a
known reference mixture of nitrogen monoxide in carbon
Column:
dioxide and comparing the actual content with the 'content
- material: stainless steel,
indicated by the analyser which has been calibrated with a
- size: 1= 2 m, 0 = 4 mm,
NOIN2 reference mixture.
- stationary phase: an appropriate molecular sieve for
chromatography (0.5 nm).
Quenching correction factor . a~ual nil~ogen monox:i~ content
Carrier gas helium for chromatography R. mdicated nitrogen monoxide content
Flow rate 60 mUmin.

Total sulfur
Temperature:
Maximum 1 ppm VIV, determined using an ultraviolet
- column: 50°C,
fluorescence analyser after oxidation of the sulfur compounds
- injection port and det«wr. 130 "C.
by heating at 1000 °C (Figure 0375.-1).
Detection Flame ionisation with methaniser.
The apparatus consists of the following:
injection Loop injector. - a system generating ultraviolet radiation with a wavelength
Adjust the injected volwnes and the operating conditions so of 210 nm, made up of an ultraviolet lamp, a collimator)
that the height of the peak due to carbon monoxide in the and a selective filter; the beam is blocked periodically by a
chromatogram obtained with the reference gas is at least chopper rotating at high speed,
35 per cent of the full scale of the recorder. - a reaction chamber through which flows the previously
Limit: filtered gas to be examined,
- carbon monoxide: not more than the area of the - a system that detects radiation emitted at a wavelength of
corresponding peak in the chromatogram obtained with 350 nm, made up of a selective filter, a photomultiplier
the reference gas (5 ppm VIV). tube and an amplifier.
Nitrogen monoxide and nitrogen dioxide Gas to be examined The substance to be examined.
Maximum 2 ppm VIV in total, determined using a Reference gas (a) Carbon dioxide RI.
chemiluminescence analyser (2.5.26). Reference gas (b) A mixture containing between
Gas to be examined The substance to be examined. 0.5 ppm VIV and 2 ppm VIV of hydrogen sulfide RI in carbon
Reference gas (a) Carbon dioxide RJ. dioxide RJ.
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1-438 Carbon Monoxide 2022
Calibrate the apparatus and set the sensitivity using reference

gases (a) and (b). Pass the gas (0 be examined through a
Carbon Monoxide
quartz oven heated to 1000 DC. Oxygen R is circulated in the (ph. Eur. monograph 2408)
oven at a tenth of the flow rate of the gas to be examined.
Measure the sulfur dioxide content in the gaseousmixture co 28.00 6JIJ-08-0
leaving the oven. PhE<I _
Water DEFINITION
Maximum 67 ppm VIV, determined using an electrolytic Gas obtained by steam reforming (catalytic oxidation) of
hygrometer (2.5.28). hydrocarbons.
Assay
Content
Infrared analyser (2.5.24).
Minimum 99.5 per cent VIV of CO.
Gas to be examined The substance to be examined. It must
This monograph applies to carbon monoxide for medicinal
be filtered to avoid stray light phenomena.
use.
Reference gas (a) Carbon dioxide RI.
CHARACTERS
Reference gas (b) A mixture containing 95.0 per cent VIV of
Appearance
carbon dioxide RJ and 5.0 per cent VIV of nurogen Rl.
Colourless, flammable gas.
Calibrate the apparatus and set the sensitivity usingreference
gases (a) and (b). Measure the content of carbondioxide in Solubility
the gas to be examined. At 20"C and at a pressure of 101 kPa, 2.266 volumes of
carbon monoxide dissolve in 100 volumes of water.
IDEl\7IFlCATION
IDENTIFICATION
Carry out either testA or B.
Comparison Ph. Bur. reference spectrum of carbon monoxide.
Comparison Ph. Eur. reference spectrnm of carball dioxide.
B. It complieswith the limits of the assay.
B. Place a glowing splinter of wood in an atmosphere of the
substance to be examined. It is extinguished. TESTS
C. Pass a stream of the substance to be examined through Carbon dioxide
ban'um hydroxide solution R. A white precipitate is fanned Gas chromatography (2.2.28).
which dissolves with effervescence in dilute autic acid R. Gas w be examined The substance to be examined"
TESTS Reference gas A mixture containing 300 ppm VIV of carball
Examine the gaseous phase. dioxide RI in carboll monoxide R.
If the us, is peiformed (}II a cylillder of gas, kup the cylillder of Column:
the subsrance to he examined at room temperature for not less than - material: stainless steel;
6 h before carryillg out the USIS. Keep the cyli"der ill the vertical - size: {= 2 m, 0 = 2 mmj
position with the outlet valve uppermost. - stationary phase: an appropriate divinylbenzene porous
polymer (149-177 urn).
Carbon monoxide
Maximum 5 ppm VIV, determined using a carbon monoxide
detector tube (2.1.6). Flow rate 30 mIlrnin.
Hydrogen sulfide Temperature:
Maximum I ppm VIV, determined using a hydrogen sulfide - column: 50 °Ci
detector tube (2.1.6). - detector: 220 'C.
Nitrogen monoxide and nitrogen dioxide
Maximum 2 ppm VIV in total, determined using a nitrogen Illje<cioIl I mL.
monoxide and nitrogen dioxide detector tube (2.1.6). Run time 3 min.
Sulfur dioxide Relative retention With reference to carbonmonoxide
Maximum 2 ppm V/~ determined using a sulfur dioxide (retention time = about 0.4 min): carbon
detector tube (2.1.6). dioxide = about 3.5.
Water vapour Lima:
Maximum67 ppm V/~ determined using a watervapour - carbon dioxide: not more than the area of the
detector tube (2.1. 6). corresponding peak in the chromatogram obtainedwith
the reference gas (300 ppm VIV).
STORAGE
Store liquefied underpressure in suitable containers Methane
complying with the legal regulations. Gas chromatography (2.2.28).
IMPURITIES
Reference gas A mixture containing 100 ppm VIV of
A. NO: nitrogen monoxide,
methane R in carbon monoxide R.
B. N0 2 : nitrogen dioxide,
Column:
C. CO: carbonmonoxide, - material: stainless steel;
D. total sulfur, - size: / = 2 m, 0 = 4 mm;
E. H2 0 : water" - stationary phase: ethylviny/benzene-dim"nylbenzene
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I copolymer R (177-250 pm).
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2022 Carbon Monoxide Intermix (5 per cent) in Nitrogen 1-439
Canier gas nitrogen for chromatography R. IMPURITIES

Flow rate 10 mUmin. Specified impurities A, B, C, D, E, F.
Temperature: A. CO2 : carbon dioxide,
- column: 95 "C; B. CH4 : methane)
- deteaor: 240 "C. C. H 2 : hydrogen,
Detection Flame ionisation. D. Ni(COJ.: nickeltetracarbonyl,
Itrie<t;on 1 mL. E. Fe(CO),: iron pentacarbcnyl,
Run time 3 min. F. H 20: water.
Retention rime Methane » about 1.8 min. -------- PIIE"
Limir.
- methane: not more than the area of the corresponding
peak in the chromatogram obtained with the reference gas
(100 ppm VIV).
Carbon Monoxide Intermix ***
Hydrogen *** ***
Gas chromatography. (5 per cent) in Nitrogen ***
Gas to be examined The substance to he examined. (Ph. Eur. monograph 2904)
Reference gas A mixture containing 300 ppm VIV of PhE" _
hydrogen for chromatography R in carbon monoxide R.
DEFINITION
Column:
A mixture containing 5 per cent V/Vof Carbon monoxide
- mouriot: stainless steel; (2408) in Low-oxygen nitrogen (1685).
- size: 1= 2 m, 0 = 2 mm;
- stationary phase: molecular sieve for chromatography Content
(149-177 urn) with a nominal pore size of 0.5 nm. 4.75 per cent VIVto 5.25 per cent V/V of carbon monoxide
(CO) in nitrogen (N2).
Comer gas argon for chromatography R.
TIlls monograph applies to carbon monoxide intermix
Flow rate 30 mUmin.
(5 per cent) in nitrogen used in me preparation of lung
Temperature: function test gas mixtures for medicinal use.
- column: 100 °Cj
- detector: 160 "C. CHARACTERS
Appearance
Colourless gas.
Itriection 1 mL.
Run time 4 min.
IDENTIFICATION
Carry out either tests A, C or tests B, C.
Relative retention With reference to carbon monoxide
(retention time = about 2.3 min): hydrogen = about 004. A. Infrared absorption spectrophotometry (2.2.2f).
Limir. Comparison Ph. Bur. reference spectrum of carbon monoxide.

- hydrogen: not more than the area of the corresponding B. It complies with the limits of me assay.
peak in the chromatogram obtained with the reference gas C. Gas chromatography (2.2.28).
(300 ppm VIV). Gas to be examined The substance to be examined.
Nickel tetracarbonyl and iron pentacarbonyl Reference gas Nitrogen RI.
Not detectable, using a detector tube having a limit of Column:
detection of 0.1 ppm VIV (2.1.6). - material: stainless steel;
Water - size: I = 2 ffi, «0 = 2 mm;
Maximum 10 ppm V/~ detennined using an electrolytic - stationary phase: molecular sieve for chromatography R
hygrometer (2.5.28). (0.5 nm).
ASSAY Carnergas helium for chromatography R.
Infrared analyser (2.5.25). Flow rate 20 mUmin.
Gas to be examined The substance to be examined, Temperature:
previously filtered to avoid stray light phenomena. - column: 80 °c;
Reference gas (a) Carbon monoxide R. - detector: 130 "C.
Reference gas (b) A mixture containing 95.0 per cent VIV of Detection Thermal conductivity.
carbon monoxide Rand 5.0 per cent V/V of nitrogen Rl. Injection 10 1'1.
Calibrate the apparatus and set the sensitivity using reference Retention time Nitrogen = about 2 min.
gases (a) and (b). Measure the content of carbon monoxide Results The principal peak in the chromatogram obtained
in the gas to be examined. with the gas to be examined is similar in retention time to
STORAGE the principal peak in the chromatogram obtained with me
Under pressure in suitable containers complying with the reference gas.
legal regulations. TESTS
Water (2.5.28)
Maximum 10 ppm VIV.
ASSAY
Infrared analyser (2.5.25).
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1-440 Carboplatin 2022
Gas to be examined The substance to be examined. It must Appearance of solution

be filtered to avoid stray-light phenomena. Solution S is dear (2.2.1) and colourless (2.2.2, Method JI).
Reference gas (a) Mixture containing 5.0 per cent VIV of Impurity B and acidity
carbo" monoxide R in nitrogen RI. Maximum 0.5 per cent, calculated as impurity B.
Reference gas (b) Nitrogen Rl. To 10 mL of solution S add 0.1 mL of phenolphthalein
Calibrate the apparatus and set the sensitivity using reference solution RJ. The solution is colourless. Not more than
gases (a) and (b). Measure the content of carbonmonoxide 0.7 mL of 0.01 M sodium hydroxide is required 10 change the
in me gas to be examined. colour of the indicator to pink.
STORAGE Related substances
As a compressed gas, in appropriate high-pressure cylinders Liquid chromatography (2.2.29).
complying with the legal regulations. Testsolution Dissolve 20.0 mg of the substance to be
examined in a mixture of equal volumes of acetonitrile Rand
LABELLING
water R and dilute to 20.0 mL with the same mixture of
The label states the nominalcontent, in per cent VIV, of solvents.
carbon monoxide in nitrogen.
IMPURITIES 200.0 mL with the mobile phase.
Specified impurities A. Golumn:
- size: 1= 0.25 m, 0 = 4.6 nun;
- 'tationaryphase: aminopropylsily/ S17i<a gd for
A. water.
_~ PhEIl
Mobile phase waterfor chromatography R, acetonitrile for
chromatography R (13:87 VII').
Flow rate 2 mUmin.
Carboplatin Injection I0 ur,
Runlime 2.5 times the retention time of carboplatin.
(Ph. Bur. monograph 1081) Relative retention With reference to carboplatin (retention
= =
time about 7 min): impurity A abouIO.3.
System suitability Test solution:
- number of theoretical plates: minimwn 5000; if necessary,
adjust the concentration of acetonitrile in the mobile
phase;
- symmetry factor: maximum 2.0; if necessary, adjust the
371.3 41575-94-4 concentration of acetonittile in the mobile phase.
Limits:
Action and use - impurity A: not more than the area of the principal peak
Platinum-containing cytotoxic. in the chromatogram obtained with the reference solution
Preparation (0.25 per cent);
Carboplatin Injection - unspuified impun'ties: for each impurity, not more than
0.4 timesthe area of the principal peak in the
PhEIl _ chromatogram obtained with the reference solution
DEFINITION (0.10 per cent);
(SP-4-2)-Diammine[cydobutan-I,I-di(carboxylato-KO)(2-)] - touil: not more than twice the area of the principal peak in
the chromatogram obtained with the reference solution
platinum.
(0.5 per cent);
Content - disregard limir. 0.2 times the area of the principal peak in
98.0 per cent 10 102.0 per cent (dried substance). the chromatogram obtained with the reference solution
CHARACTERS (0.05 per cent),
Appearance Chlorides (2.4.4)
Colourless, crystalline powder. Maximum 100 ppm.
Solubility Dissolve 0.5 g in water R, heating slightly if necessary, and
Sparingly soluble in water, very slightly soluble in acetone dilute to 20 mL with the same solvent. Filter if necessary.
and in ethanol (96 per cent). Dilute 10 mL of this solution to 15 mL with water R.
mp Prepare the standard using 5 mL of chloride standard solution
About 200 °C, with decomposition. (5 ppm GI) R.
IDENTIFICATION Ammonium (2.4.1, Method B)

Maximum 100 ppm, determined on 0.20 g.
Prepare the standard using 0.2 mL of ammonium standard
Comparison Ph. Bur. reference spectrum of carboplalin.
solution (100 ppm NH.,) R.
TESTS Loss on drying (2.2.32)
Solution S Maximum 0.5 per cent, determined on 1.000 g by drying in
Dissolve 0.25 g in carbon dioxide-free water R and dilute to an oven at 105°C.
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2022 Carboprost Trometamol 1-441
ASSAY Related substances

Use the residue obtained in the test for loss on drying. Ignite Liquid chromatography (2.2.29).
0.200 g of the residue to constant mass at 800 ± 50°C. Test solution Dissolve 15.0 mg of the substance to be
1 mg of the residue is equivalent to 1.903 mg of examined in a mixture of 23 volwnes of acetonitrile Rand
C6HI2Nz04Pt. 77 volumes of water for chromatography R and dilute to
STORAGE 10.0 mL with the same mixture of solvents.
Protectedfrom light. Reference solution (a) Dissolve 15.0 mg of corboprost
trometamol CRS (containing impurity A) in a mixture of
IMPURITIES 23 volumes of ocetoninile Rand 77 volumes of waterfor
Specified impurities A, B. chromatography R and dilute to 10.0 mL with the same
mixture of solvents.
Reference ,00ution (b) Dilute 1.0 mL of reference solution (a)
and 0.15 mL of (l5R)-15-m,thylprostagfandin F,. R
(impurity B) to 100.0 mL with a mixture of 23 volumes of
acetonitrile Rand 77 volumes of walerfor chromatography R.
A. (SP-4-2)-diamminedichloridoplatinum(lI) (cisplatin), Reference solution (c) Dilute 2.0 mL of the test solution to
20.0 mL with a mixture of 23 volwnes of acetonitrile Rand
()<Co,H 77 volumes of waterfor chromatography R. Dilute 2.0 mL of
CO,H this solution to 20.0 mL with a mixture of 23 volumesof
acetonitrile Rand 77 volwnes of walerfor chromatography R.
B. cyc1obutane-l,l-dicarboxylic acid. Column:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" - size: 1= 0.15 m, 0 = 4.6 nun,
- stationary phase: octade<ylsilyl ,ilicagelfor
chromatography R1 (5 I'm) with a pore size of 8-10 run
and a carbonloading of 12-19 per cent.
Carboprost Trometamol Mobile phase Mix 23 volumes of acetonitrile RJ and
77 volumes of a 2.44 gIL solution of sodium dihydrogen
(ph. Eur. monograph 1712) phosphate R in waterfor chromatography R previously adjusted
to pH 2.5 with phosphoric acid R.
~OH
(Nt<, Injection 20 ~L.
OH Run time 1.3 times the retention time of carboprost.
Relative retention Wirh reference to carboprost (retention
C"Ht,NO. 489.7 58551-69-2 = =
time about 80 min): impurity B about 0.85;
Action and use Identification of impurities Use the chromatogram obtained
Prostaglandin (pGF,J analogue. with reference solution (a) and the chromatogram supplied
PhE" _ with casboprosi trometomd CRS to identify the peak due to
impurity A.
DEFINITION System suitabt7ity:
2-Amino-2-(hydroxymethyl)propane-I,3-diol (5Z)-7- - resolution: minimum 3.4 between the peaks due to
[(IR,2R,3R,5S)-3,5-dihydroxy-2-[(IE,3S)-3-hydroxy-3- impurity Band carboprost in the chromatogram obtained
methyloct-I-enyl]cyclopentyl]hept-5-enoate «15S)-15- with reference solution (b);
methyl-PGF2 ) . - peak-to-valley ratio: minimum 3.0, where Hp = height
Content above the baseline of the peak due to impurity A and
94.0 per cent to 102.0 per cent (anhydrous substance). H v = height above the baseline of the lowest point of the
CHARACTERS curve separating this peak from the peak due to
impurity B in the chromatogram obtained with reference
Appearance
solution (a).
White or almostwhite powder.
Limits:
Solubility
- impun"ty A: not more than 3 times the area of the
Soluble in water.
IDENTIFICATION reference solution (c) (3.0 per cent),
A. Specific opticalrotation (see Tests). - impun"lY B: not more than the area of the principal peakin
B. Infrared absorption spectrophotometry (2.2.24). the chromatogram obtained with reference solution (c)
(1.0 per cent),
Comparison Ph. Bur. reference spectrum of carboprost
tromeuzmol. - unspecified impurities: for each impurity, not more than
TESTS chromatogram obtained with reference solution (c)
Specific optical rotation (2.2.7) (0.10 per cent),
+ 18 to + 24 (anhydrous substance). - total: not more than 4 times.the area of the principal peak
Dissolve 0.100 g in ethanol (96 percent) R and dilute to in me chromatogram obtained with reference solution (c)
10.0 mL with the same solvent. (4.0 per cent),
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1-442 Carmellose 2022
- disregard limit: 0.05 times the area of the principal peak in Solubility
the chromatogram obtained with reference solution (e) Practically insoluble in anhydrous ethanol. It swells with
(0.05 per cent). water to form a suspension and becomes viscid in I M
Water (2.5.32) sodium hydroxide.•
Maximwn 0.5 per cent, determined on 50 mg. IDENTIFICATION
ASSAY A. pH (2.2.3): 3.5 to 5.0.
Liquid chromatography (2.2.29) as described in the test fer Suspend 1.0 g in 100 mL of carbon dioxUk-Jre' water R.
related substances with the following modifications. B. Infrared absorption spectrophotometry (2.2.24).
Mobile phase Mix 27 volumes of aatonilriJe Rl and Comparison carmellose CRS.
73 volumes of a 2.44 gIL solution of sodium dihydrogen
TESTS
phosphate R in water for chromatography R previously adjusted
to pH 2.5 with phosphoric acidR. Chlorides
InjtXtWn Test solution and reference solution (a).
Shake 0.8 g with 50 mL of water R, dissolve in 10 mL of
Run time 1.2 times the retention time of carboprosr. 1 M sodium hydroxide and dilute to 100 mL with water R.
Retention time Carboprost = about 29 min. Heat on a water-bath a mixture of 10 mL of dilute nitric
Calculate the percentage content of C:!5Ht,NOS using the acidRand 20 rnL of this solution until a flocculent
declared content of carboprost trometamd CRS. precipitate is produced. Cool, centrifuge andtake out the
STORAGE supernatant. Wash the precipitate with 3 quantities, each of
10 mL, of water R, centrifuging each time. Combine the
At a temperature below -15 0 C.
supernatant and the washings and dilute to 100 mL with
IMPURITIES water R. To 25 mL of this solution add 6 mL of dilute ni'ric
Specified impurities A, B. acidR and dilute to 50 mL with water R (test solution).
Prepare the reference solution in the same manner, using
0.40 mL of 0.01 M hydrochloric acid. Add I mL of SIlver
nitrate solution R2 to the test solution and the reference
solution. Allow to stand protected from light for 5 min.
Any opalescence in the test solution is not moreintense man
that in the reference solution.
Sulfates
A. (5E)-7 -[( IR,2R,3R,5S) -3,5-dihydroxy-2- [(IE, 3S)-3-
hydroxy-3-methyloct-I-enyl]cyclopentyl]hept-5-enoic acid,
Shake 0.40 g with 25 mL of water R, dissolve in 5 mL of
1 M sodium hydroxide and add 20 mL of water R. Heat this
solution with 2.5 mL of hydrochlork acidR in a water-bath
until a flocculent precipitate is produced. Cool, centrifuge,
and takeout the supernatant. Wash theprecipitate with
3 quantities, each of 10 mL, of water R J centrifuging each
time. Combine the supernatant and the washings, and dilute
B. (5Z)-7-[(IR,2R,3R,5S)-3,5-dihydroxy-2-[(1E,3R)-3- to 100 mL with waterR. Filter, and discard the first 5 mL of
hydroxy-3-methyloct-I-enyl]cyclopentyl]hept-5-enoic acid. the filtrate. To 25 mL of the filtrate add I mL of dilute
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII hydrochloric acid R and dilute to 50 mL with water R (test
solution). Prepare the reference solution in the same manner,
using 1.5 mL of 0.005 M sulfum acid. Add 2 mL of a
120 gIL solution of barium chloride R to the testsolution and
Carmellose1 the reference solution. Mix and allow to stand for 10 min.
The white turbidity produced in the test solution is not
(ph. Eur. monograph 2360) thicker than that in the reference solution.
9000-11-7 Maximum 8.0 pet cent, determined on 1.000 g by drying in
Action and use an oven at 105 °C for 4 h.
Excipient; bulk laxative, Sulfuted ash (2.4.14)
, Maximum 1.5 per cent (dried substance), determined on
PIIEII _
1.0 g.
DEFINITION .STORAGE
Carboxymethylether of cellulose. In an airtight container.•
Partly D-carboxymethylated cellulose. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII
.CHARACTERS
Appearance
I 17Iif monograph has undergone phannawpoeial hannonisatWti.

See chapter 5.8 Pharmacopoeial harmonisation.
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2022 Carmellose Sodium 1-443
Carmellose Calcium 1 Heat 20 mL of solution S with I mL of hydrochloric acidR

on a water-bath until a flocculent precipitate is produced.
(ph. Eur. monograph 0886) Cool, centrifuge and separate the supernatant. Wash the
precipitate with 3 quantities, each of 10 ml., of distilled
9050-04-8 water R, centrifuging each time. Combine the supernatant
and the washings and dilute to 100 mL with distilled water R.
Action and use To 25 mL add I mL of dilute hydrochloric acid R and dilute
Excipient in pharmaceutical products; bulk laxative.
to 50 mL with dis'iBed wat<r R.
Pl>E" _ Loss on drying (2.2.32)
DEFINITION
Calcium salt of partly o-carhoxymethylated cellulose.
Calcium salt of a polycarboxymethyl ether of cellulose. Sulfated ash (2.4.14)
10.0 per cent to 20.0 per cent, determined on 1.0 g of the
.CHARACTERS dried substance.
Appearance
White or yellowish-white powder, hygroscopic after drying. .STORAGE
In an airtight container.•
Solubility
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Practically insoluble in acetone, in ethanol (96 per cent) and
in toluene. It swells with water to Conn a suspension."
IDENTIFICATION
•**"*•••
A. Shake 0.1 g thoroughly with 10 mL of wow R. Add 2 mL
of a 42 gIL solution of sodium hydroxide R and allow to stand
Carmellose Sodium
for 10 min (solution A). Dilute I mL of solution A to 5 mL (Ph. Eur. monograph 0472) *.*
with wow R. To 0.05 mL of this solution add 0.5 mL of a
0.5 gIL solution of chromoeopic acid, sodium salt R in a 9004-32-4
75 per cent m/m solution of suljunc acidR and heat on a
water-bath for 10 min. A reddish-violet colour develops. Action and use
Excipient; bulk laxative.
B. Shake 5 mL of solution A obtained in identification test A
with 10 mL of acetone R. A white, flocculent precipitate is Preparation
produced. Carmellose Sodium Eye Drops
C. Shake 5 mL of solution A obtained in identification test A Pl>E" ~_
with I mL of/en;c chloride solution RI. A brown, flocculent

DEFINITION
CarboxymethylceUulose sodium. Sodium salt of a partly
D. Ignite 1 g to ash and dissolve the residue in a mixture of
O-carboxymethylated cellulose.
5 mL of acetic acid R and 10 mL of wow R. Filter if
necessary and heat the filtrate to boiling. Cool and neutralise Content
with di/ute ammonia RI. The solution gives reaction (a) of 6.5 per cent to 10.8 per cent of sodium (Na) (dried
calcium (2.3.1). substance).
TESTS CHARACTERS
Solution S Appearance
Shake 1.0 g with 50 mL of dis'illed wow R, add 5 mL of White or almost white, granular powder, hygroscopic after
dilute sodium hydroxide solution R and dilute to 100 mL with drying.
distiBed water R. Solubility
AlkalInity Practically insoluble in acetone, in anhydrous ethanol and in
Shake 1.0 g thoroughly with 50 mL of carbon dwxide-free toluene. It is easily dispersed in water giving colloidal
water R and add 0.1 mL of phenolphthalein solurion R. No red solutions.
colour develops. IDENTIFICATION
CbIorides (2.4.4) A. To 10 mL of solution S (see Tests) add I mL of copper
.M aximum 0.36 per cent. sulfate solution R. A blue, cotton-like precipitate is formed .
Heat 28 mL of solution S with 10 mL of dilute nimc acid R B. Boil 5 mL of solution S for a few minutes. No precipitate
on a water-bath until a flocculent precipitate is produced. is formed.
Cool, centrifuge and separate the supernatant. Wash the C. To the residue obtained in the determination of the
precipitate with 3 quantities, each of 10 mt, of water R, sulfated ash, add I mL of hydrochloric acid R and evaporate
centrifuging each time. Combine the supernatant and the on a water-bath. Take up the residue in 20 mL of water R.
washings and dilute to 100 mL with waW R. To 25 mL add The solution gives the reactions of sodium (2.3.1).
6 mL of dilute nuricacidR and dilute to 50 mL with waterR.
TESTS
Dilute 10 mL of the solution to 15 mL with water R.
Solution S
Sulfates (2.4.13) Sprinkle a quantity of the substance to be examined
Maximum 1 per cent. equivalent to 1.0 g of the dried substance onto 90 mL of
carbon dioxide-free waterR at 40 °C to 50°C stirring
vigorously. Continue stirring until a colloidal solution is
obtained, cool and dilute to 100 mL with carbon dioxide-free
I This m()tIQgraph has undergone plulr7nacopoeia/ harmonisation:
water R.
See clutpler 5.8 PharmaC<JpoeiaJ hannonisalion.
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1-444 Carmellose Sodium 2022
Appearance of solution - for a productof low viscosity, the concentration of the

Solution S is not more opalescent than reference solution to be used and the viscosity in millipascal
suspension III (2.2.1) and not more intensely coloured than seconds.
reference solution Y6 (2.2.2, Aletlwd11). _~ PhE<I
pH (2.2.3)
The pH of solution S is 6.0 to 8.0.
Viscosity
75 per cent to 140 per cent of the valuestated on the label. Low-substituted Carmellose ******
**••**
While stirring, introduce a quantity of the substance (Q be Sodium
examinedequivalent to 2.00 g of the dried substance into
50 mL of water R heated to 90°C. For a product of low (Ph. Eur. monograph 1186)
viscosity, use, if necessary, the quantity required to give the 9051J-32-4
concentration indicated on the label. Allow to cool, dilute to
100.0 mL with water R and stir until dissolution is complete. Action and use
Determine me viscosity (2.2.10) using a rotating viscometer Excipient in pharmaceutical products; bulk laxative.
at 20°C and a shear rateof 10 s-I, If it is impossibleto PhEIl ~
obtain a shear rateof exactly 105- 1, use a shearrate slightly

higher and a rate slightly lower and interpolate. DEFINITION
Sodium glycolate Low-substituted sodiwn carboxymethylceUulose. Sodium salt
Maximum 0.4 per cent (driedSUbstance). of a partly D-(carboxymethylated) cellulose.
Test solution Place a quantity of the substance to be Content
examinedequivalent to 0.500 g of dried substance in a 2.0 per cent to 4.5 per cent of sodium (Na) (dried
beaker. Add 5 mL of acetic acidR and 5 mL of waterR. Stir substance).
until dissolution is complete (about 30 min). Add 80 mL of CHARACTERS
acetone Rand 2 g of sodium chloride R. Filter through a fa" Appearance
filter paper impregnated with acetone R into a volumetric White or almost white powder or short fibres.
flask, rinse the beaker and filter with acetone R and dilute the
filtrate (Q 100.0 mL with the same solvent. Allow to stand for Solublllty
24 h without shaking. Use the clear supernatant. Practically insoluble in acetone, in anhydrous ethanol and in
toluene. It sweUs in water to form a gel.
Reference solution In a volumetric fiaskJ dissolve 0.310 g of
glycolic acid R, previously dried in a desiccator (2.2.32), in IDENTIFICATION
water R and dilute to 1000.0 mL with the same solvent. A. Shake I g with 100 mL of a 100 gIL solution of sodium
Place 5.0 mL of this solution in a volumetric flask, add 5 mL hydroxide R. A suspension is produced.
of acetk add R and allow to stand for about 30 min. B. Shake I g with 50 mL of walei' R. Transfer I mL of the
Add 80 mL of acetone Rand 2 g of sodium chloride Rand mixture to a test tube, add 1 mL of water Rand 0.05 mL of
dilute to 100.0 mL with acetone R. a freshly prepared 40 gIL solution of a-naphmol R in
Place 2.0 mL of each solution in a separate 25 mL methanol R. Incline the test tube and add carefully 2 mL of
volumetric flask. Heat on a water-bath to eliminate acetone. sulfuric add R down the side so that it forms a lower layer.
Cool to room temperature and add 5.0 mL of A reddish-purple colour develops at the interface.
2,7-<1ihydroxynaphlhaiene solution R to each flask. Shake and C. Sulfated ash (2.4.14) (see Tests).
add 15.0 mL of 2,7-dihydroxynaphlhalene solulwn R. Close the D. To the residue obtained in the determination of the
flasks with aluminiwn foil and heat on a water-bath for sulfated ash, add I mL of hydrochloric acidR and evaporate
20 min. Cool under running water and dilute to 25.0 mL on a water-bath. Take up the residue in 20 mL of water R.
with sulfuric acid R. Within 10 min, transfer 10.0 mL of each The solution gives reaction (a) of sodium (2.3.1).
solution to a flat-bottomed tube. Examine the solutions
viewing vertically. The test solution is not more intensely TESTS
coloured than the reference solution. pH (2.2.3)
6.0 to 8.5.
Chlorides (2.4.4)
Maximum 0.25 per cent. Shake I g with 100 mL of carban dioxide-free walei' R for
5 min. Centrifuge.
Sodium chloride and sodium glycolate
.; Maximum 0.5 per cent (dried substance) for the sum of the
percentage contents.
an oven at 105°C.
Sodium chloride Place 5.00 g in a 250 mL conical flask, add
Sulfuted ash (2.4.14) 50 mL of walei' Rand 5 mL of "rang hydrogen peroxide
20.0 per cent to 33.3 per cent, determined on 1.0 g using a solution R and heat on a water bath for 20 min, stirring
mixture of equalvolumes of sulfuric acid R and water Rand occasionally to ensure total hydration. Cool, add 100 mL of
calculated with reference to the dried substance. These limits water Rand 10 mL of niuic acid R. Titrate with 0.05 Atl -siluer
correspond to a content of 6.5 per cent to 10.8 per cent of nitrate determining the end-point potentiometrically (2.2.20)
sodium (Na). using a silver-based indicator electrode and a double-junction
LABELLING reference electrode containing a 100 gILsolution of potassium
The label states: nitrate R in the outer jacketand a standard filling solution in
- the viscosity in millipascal seconds for a 20 gIL solution; the inner jacket.
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2022 Carmustine 1-445
1 mL of 0.05 M silver nitrate is equivalent to 2.922 mg FUNCTIONALITY-RELATED CHARACTERISTICS

of NaCI. This section provides infonnation on characteristics that are
Sodium glycolate Place a quantity of the substance to be recognised as being relevant control parameters for one ormore
examined equivalent to 0.500 g of the dried substance in a functions of the substance when usedas an excipient (see chapter
beaker. Add 5 mL of glacial acetic acid Rand 5 mL of 5.15). Some of the characteristics described in the Funcckmality-
water R and stir £0 ensure total hydration (about 30 min). related characteristics section may also bepresent in the mandatory
Add 80 mL of acetone Rand 2 g of sodium chloride R. Stir for part of the monograph since they also represent mandawryquality
several minutes to ensure complete precipitation of the criteria. In such cases, a cross-reference to the tests described in the
carboxymethylceUulose. Filter through a fast filter paper mandatory part is included in the FunctUmaHty-relaced
impregnated with acetone R into a volumetric flask, rinse the characteristics section. Control of the characteristics can conm'bule
beaker and filter with acetone R and dilute the filtrate to to ,he quality of a medicinal product Iry impnnling the consistency
100.0 mL with the same solvent. AUow to stand for 24 h of the manufacturing proem and the peiformance of the medicinal
without shaking. Use the clear supernatant as the test produa during use. Where control methods are cued, they are
solution. recognised as being suitable for the purpose, but othermethods can
Prepare the reference solutions as follows: in a 100 mL also be used. Wherever results for a panicular characteristic are
volumetric flask, dissolve 0.100 g of glycolic acid R, previously reported, the control method must he indicaud.
dried in a desiccator (2.2.32), in water R and dilute to Thefollowing characteristic may be relevant for low-substituted
100.0 mL with the same solvent. Transfer 0.5 mL, 1.0 mL, carmellose sodium usedas disintegrant.
1.5 mL and 2.0 mL of the solution to separate volumetric Settling volume
flasks; dilute the contents of each flask to 5.0 mL with 15.0 mL to 35.0 mL.
water R, add 5 mL of glacial acetic acid R, dilute to 100.0 mL In a 100 mL graduated cylinder, place 20 mL of
with acetone R and mix. 2-propanol R, add 5.0 g of the substance to be examined and
Transfer 2.0 mL of the test solution and 2.0 mL of each of shake vigorously. Dilute to 30 mL with 2-propanol R then to
the reference solutions to separate 25 mL volumetric flasks. 50 mL with water R and shake vigorously. Within 15 min,
Heat the uncovered flasks in a water-bath to eliminate the repeat the shaking 3 times. Allow to stand for 4 hand
acetone. Allow to cool and add 5.0 mL of determine the volume of the settled mass.
2.7-dihydroxynaphthalcne solution R to each flask. Mix, add a _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw
further 15.0 mL of 2,7-dihydroxynaphthalcne solution Rand
mix again. Close the flasks with aluminiwn foil and heat in a
water-bath for 20 min. Cool and dilute to 25.0 mL with
sulfuric acid R.
Measure the absorbance (2.2.25) of each solution at 540 nm.
Carmustine
Prepare a blank using 2.0 mL of a solution containing (Ph. Bur. monograph 1187)
5 per cent VIVeach of glacialacetic acid R and water R in
aceUme R. Prepare a standard curve using the absorbances
obtained with the reference solutions. From the standard
curve and the absorbance of the test solution, determine the
mass GJ in milligrams, of glycolic acid in the substance to be
examined and calculate the content of sodium glycolate from
the following expression: C,H,CI,N,O, 214.1 154-93-8
lOx 1.29xa Acdon and use
(100 - b)m Cytotoxic alkylating agent.
1.29 factor converting glyeolic acid to sodiwn glycolate. PhEw _
b loss on drying as a percentage.
m mass of the substance to be examined. in grams. DEFINITION
N,N'-Bis(2-chloroethyl)-N-nitrosourea.
Water-soluble substances Content
Maximum 70.0 per cent. 98.0 per cent to 102.0 per cent (anhydrous substance).
Disperse 5.00 g in 400.0 mL of water R and stir for 1 min
CHARACTERS
every 10 min during the first 30 min. Allow to stand for 1 h
Appearance
and centrifuge, if necessary. Decant 100.0 mL of the
Yellowish, granular powder.
supernatant onto a fast filter paper in a vacuum filtration
funnel, apply vacuum and collect 75.0 mL of the filtrate. Solubility
Evaporate to dryness and dry the residue at 100-105 "C Very slightly soluble in water, very soluble in methylene
for 4 h. chloride, freely soluble in anhydrous ethanol.
Loss on drying (2.2.32) mp
Maximum 10.0 per cent, determined on 1.000 g by drying in About 31 "C, with decomposition.
an oven at 105 "C. IDENnFlCATION
Sulfated ash (2.4.14) Infrared absorption spectrophotometry (2.2.24).
6.5 per cent to 13.5 per cent (dried substance), Preparation Examine the melted substance prepared as a
corresponding to a content of 2.0 per cent to 4.5 per cent of film.
Na. Comparison carmustine CRS.
Use 1.0 g with a mixture of equal volumes of sulfim'c acid R
and water R.
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1-446 Carmustine 2022
TESTS Tim. Mobile phase A Mobile phase B

(min) (per cent VIV> (per cent VIJI)
Impurity B
Thin-layer chromatography (2.2.27). 0·10 8' 15
10 ~ 60 85 ..... )0 15 ...... 70
examined in 0.5 mL of methanol R.
Reference solution (a) Dissolve 30.0 mg of chloroethylamine FI= rau 1.2 mllmin.
hydrochloride R (impurity B) in 25 mL of methanol Rand Detection Spectrophotometer at 230 run and, for
dilute to 50.0 mL with the same solvent. impurity A, at 205 run.
Referencesolution (b) Dissolve 0.20 g of the substance to be Autosampler Set at 5°C.
examined in 0.5 mL of reference solution (a). lnjecu"on 20 J.1L of the test solution and reference
Piau TLC silica gelpIau R. solutions (a), (c) and (d).
Mobile phase: Identification of impun·ties Use the chromatogram obtained
- mobile phaseA: ethyl alX!taU R; with reference solution (c) to identify the peakdue to
- mobile phase B: methanol R, ethyl awau R (30:10 VIV). impurity A.
Applica,ion 1 ~L. Relativeretention Withreference to carmustine (retention
Deoelopment Over 2/3 of the plate with mobile phase A, =
time about 36 min): impurity A = about 0.3.
allow to dry in airfor 5 min; over 1/3 of me platewith Systemsuitability Reference solution (d):
mobilephase B, allow to dry in air for 10 min. - resolution: minimum 5.0 between the peaks due to
Detection Spray with diethylamine R and heat at 105 °C for impurity A and carmustine.
20 min; allow to cool and spray with si/~r nimue solution R2. Calculation of percentage contents:
Expose to ultraviolet light at 365 run until brown to black - for impurity A, use the concentration of impurity A in
spots appear and examine in daylight. reference solution (c);
Retordation faaors Impurity B = about 0.2; - for impurities other than AJ use the concentration of
carmustine in reference solution (a).
carmustine = about 0.9.
System suitability Reference solution (b): Limits:
- the chromatogram shows 2 dearly separated spots. - impurity A: maximum 0.2 per cent;
Limit: 0.10 per cent;
- impun"ty B: any spot due [0 impurity B is not more intense
thanthe spot in the chromatogram obtained with - reporting threshold: 0.05 per cent.
reference solution (a) (0.1 per cent).
Water (2.5.12)
Related substances maximum 1.0 per cent, determined on 0.500 g.
Liquid chromatography (2.2.29). Cany ou' the us, protected
from lighc Prepare the solutions immediauiY before use. ASSAY
Solven, mixture ethanol (96 per cen,) R, wau, R (10:90 VIV). Dissolve 0.100 g in 30 rnL of anhydrous ethanol R and dilute
to 100.0 rnL with waue R. Dilute 3.0 mL of the solution to
100.0 rnL with waur R. Measure the absorbance (2.2.25) at
examined in 1 rnL of ethanol (96 per cen!> R and dilute to
the absorption maximum at 230 nm.
Calculate the content of C,H,Cl,N,O, taking the specific
Reference so/utum (a) Dilute 1.0 mL of the test solution to
absorbance (0 be 270.
100.0 rnL with the solvent mixture. Dilute 1.0 rnL of this
solution to 10.0 mL with the solventmixture. STORAGE
Reference sdution (b) Dissolve J0.0 mg of cannustine In an airtight container, protected from light, at a
impnn·'Y A CRS in ethanol (96 per cenO R and dilute to temperature of 2 °C to 8 °C.
10.0 mL with the same solvent. IMPURITIES
Reference solution (c) Dilute 1.0 rnL of reference solution (b) Specified impurities A, B.
to 25.0 rnL with the solvent mixture. Dilute 1.0 mL of this
Reference souuion (d) To 1 mL of the test solution add
1 rnL of reference solution (b) and dilute to 100 rnL with the
solvent mixture.
Column: A. N,N'-bis(2-chloroethyl)urea,
- size: / ;;;; 0.25 m, 12) ;;;; 4.6 mm;
- stationary phase: base-deoaiooud end-eapped octadecylsilyl c,'-./" NH,
sihea gelfor chromatography R (5 pm).
Mobile phase: B. z-chloroethan-t-emlne.
- mobile phase A: dissolve 2.1 g of pa'assiwn dihydrogen _________ ~ ""E'"
phosphore R in 1000 mL of waue for chromatography R,
add 1 rnL of methylamine R and adjust to pH 3.5 with
phosphoric acid R;
- mobile phase B: acetonitrile Rl,
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2022 Carrageenan 1-447
Acid value
Carnauba Wax 2 to 7.
(Ph. Ellr. monograph 0597) To 2.000 g (m g) in a 250 mL conicaillask fined with a
reflux condenser add 40 mL of xylene R and a few glass
8015-86-9
beads. Heat with stirring until the substance is completely
Action and use dissolved. Add 20 mL of ethanol (96 percenO R and I mL of
Excipient. bromothymol blue solution R3 and titrate the hot solution with
0.5 l\rl alcoholic potassium hydroxide until a greencolour
POE« _ persisting for at least 10 s is obtained (n, mL). Carry out a
blank test (n2 mL). Calculate the acid value using the
DEFINITION
Purified wax obtained from the leaves of Copemicia unfera
Mart. 28.05(n, - n,)
CHARACTERS m
Appearance
Pale yellow or yellowpowder, flakes or hard masses. Saponification value
78 to 95.
Solubility
Practically insoluble in water, soluble on heating in ethyl To 2.000 g (m g) in a 250 mL conicaillask fined with a
acetate and in xylene, practically insoluble in ethanol reflux condenser add 40 mL of xylene R and a few glass
(96 per cent). beads. Heat with stirring until me substance is completely
dissolved. Add 20 mL of ethanol (96 per cenO Rand 20.0 mL
Relative density of 0.5 M alcoholic potassium hydroxide. Boil under a reflux
About 0.97. condenser for 3 h. Add I mL of phenolphthalein sdtuion RI
IDENTIFICATION and titrate the hot solution immediately with
Thin-layer chromatography (2.2.17). 0.5 M hydrochlori< acid until the red colour disappears.
Test solution Dissolve 0.10 g of the substance to be Repeatthe heating and titration until the colourno longer
examined with heating in 5 mL of chloroform R. Use the reappears on heating (n3 mL). Carry out a blank test
wann solution. (n4 mL). Calculate the saponification value using the
Reference sduuon Dissolve 5 mg of memhol R, 5 1'1. of
menthyla<etate Rand 5 mg of thymolR in 10 mL of 28.05(n, - n,)
tduene R. .
m
Plate TLC sitica gelplate R.
Mobile phase ethylautate R, chloroform R (2:98 VIV). Total ash (2.4.16)
Application 30 ~L of the test solution and 10 1'1. of the Maximum 0.25 per cent, determined on 2.0 g.
reference solution as bands 20 nun by 3 nun. STORAGE
Deoelopmeni Over 1/2 of the plate, Protected from light.
Drying In air. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POE«
Detection Spray with a freshly prepared 200 gIL solution of

phosphomolybdic acidR in ethanol (96 percent) R (about
10 mL for a 20 cm plate). Heat at 100-105 "C for
Carrageenan ***
*** ***
10-15 min.
Results The chromatogram obtained with the reference
solutionshows in the lower part a dark blue zone (menthol), (Ph. Ellr. monograph 2138) ***
above thiszone a reddish zone (thymol) and In the upper POE« _
part a dark blue zone (menthyl acetate). The chromatogram
obtainedwith the test solutionshows a large blue zone DEFINITION
(triacontanol :::; melissyl alcohol) at a levelbetween the Polysaccharides extracted from different Rhodophyceae with
thymol and menthol zones in the chromatogram obtained boiling water or aqueous alkali solutions. Carrageenan is
with the reference solution. Further blue zones are visible in separated by alcoholprecipitation, potassium chloride
the upper part of the chromatogram obtained with the test precipitation, gel pressing, drumdrying or freezing.
solution, at levelsbetween those of the menthyl acetate and The alcohol used during separation and purification is
thymol zones in the chromatogram obtained with the generally 2-propanol. The main components are potassium,
reference solution; above these zones, further zones are sodium, calcium or magnesium salts of the sulfate esters of
visible in the chromatogram obtained with the test solution; D-galaetose and 3,6-anhydro-D-galaetose copolymers. They
the zone with the highest RF value is very pronounced. exist in different proportions depending on the biological
A number of faint zones are visible below the triacontanol origin of the polymer.
zone and the point of application is coloured blue. The prevalent copolymers are designated as K-, 1- and
A-carrageenan.
TESTS
Melting point (2.2.15) CHARACTERS
80°C to 88 "C. Appearance
l\rlelt the substance to be examined carefully on a water-bath Yellowish, brownish, or white or almost whitepowder.
before introduction into the capillary tubes. Allow the tubes Solubility
to stand in the refrigerator for 24 h or at 0 "C for 2 h. Soluble in water giving a viscous or colloidal solution,
insoluble in organic solvents.
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1-448 Carteolol Hydrochloride 2022
IDENTIFICATION Ash insoluble in hydrochloric acid (2.8.1)

A. Prepare a 20 gIL dispersion and heat in a water-bath at Maximum 2.0 per cent.
80 "C. Mix 1 volume of this solution and about 4 volumes of FUNCTIONALITY-RELATED CHARACfERISTICS
waterR and add 2-3 drops of a 0.5 gIL solution of methylene This section provides £nfonnation on characteristics that are
blueR in ethanol (96 per cenlj R. A blue precipitate is formed, recognised as being relevant control parameters for one or more
B. Infrared absorption spectrophotometry (2.2.24). functions of the substance when usedas an excipient (see chapter
Preparation Prepare a 2 gIL solution of the substance to be 5.15). Same of the characteristics described in the Functionali'?r
examined; cast 4.5 mL of the solution into a plastic flat- related characteristics section may also bepresent in the mandatory
bottomed weighing boat about 35 mm in diameter and aUow part of the monograph since they also represent mandatory quall'ty
to dry completely in an air-flow oven at 60°C until a film, criteria. In such cases, a cross-reference to the tests descn'bed in the
about 10 ~m thick, is obtained (about 4 h). mandatory part is included in the Funcuonoliiy-rdaud
Carrageenan has strong, broad absorption bands, typical of characteristics section. Control of the characteristics can contribuu
all polysaccharides, in the 1000-1100 ern" region. UJ the quality of a medicinal product by improving the consistency
Absorption maxima are 1065 cm! and 1020 cm-l for gelling of the manufacturing process and the perfonnance of the medicinal
and non-gelling types, respectively. Other characteristic product during use. Where control methods are ciud, they are
absorption bands and their intensities relative to the recognised as being suitable for the purpose, but other methods can
absorbance at 1050 em"! are shown in Table 2138.-1. also be used. Wherever results for a particular characteristic are
reponed, the control method must be indicated.
Table 2138.-1. - Characteristic obsotpuon bandsfor carrageenan The following charaaeriuia may be relevant for CQmlgeenan used
idenufication by infrared absorption spearopholOmetry as viscosity-increasing agent.
Absorbance relative to the Gel formation
Wavenumber absorbance at 1050 cm-l Prepare a 20 gIL dispersion and heat in a water-bath at
MolecuJar structure
(em-I)
K , • 80 °C (solution A). Allow to cool, it becomes more viscous
upon cooling and may form a gel.
1220-1260 Ester sulfate 0.7 - 1.2 1.2 -2.0 1.4 - 2.0
To 10 mL of solution A, while still hot, add 4 drops of a
3,6-Anhydro-D- 100 gIL solution of potassium chloride R, mix and allow to
928 - 933 0.3 - 0.6 0.2 - 0.5 :$. 0.2
galactose cool. A 'brittle' gel indicates a carrageenan of a
predominantly x-type; an 'elastic' gel indicates a
840 - 850 Galactoee-l-suffate 0.3·05 0.2·0.4 -
predominantly i-type; if the solution does not Conn a gel, the
825 - 830 Galactose-z-sajfate - - 0.2 - 0.4 carrageenan is of a predominantly A-type; a weak or pourable
gel obtained on cooling indicates a carrageenan comprising a
810 - 820 Galactose-S-sulfate - - 0.1 - 0.3
mixture of x- and A-types, which can be confirmed by
3,6-Anhydro-o- infrared absorption spectrophotometry.
800·80j
gaiactose-2-sulfBte
50.2 0.2 - 0.4 -
Viscosity
(see Tests).
Table 2138.-1 shows 'absorbance ratios corresponding to _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
copolymer types isolated from carrageenans. Native extracts
of carrageenan from most cold-water seaweed species
comprise IC-, 1- and A-StruCtures from the natural mix of
haploid and diploid plants. Such extracts exhibit, in practice,
the characteristic absorbance peaks of 1<:-, 1- and A-structures
Carteolol Hydrochloride
and yield absorbance ratios somewhere between the above-
mentioned individual ranges.
TESTS
and eoanliomer
Viscosity (2.2.1(J)
Minimum 5 mpa-s. Heat a 15 gIL dispersion (dried • Hel
substance) at 80 °C for at least 15 min to dissolve.
Compensate for any loss of water by evaporation, allow to
cool to 75 °C and cany out the test at this temperature. 328.8 51781-21-6
Arsenic (2.4.27)
Action and use
Maximum 3.0 ppm.
Bera-adrenoceptor antagonist.
Cadmium (2.4.27)
Preparation
Maximum 2.0 ppm.
Carreolol Eye Drops
Lead (2.4.27)
PhE" _
Maximum 5.0 ppm.
Mercury (2.4.27) DEFINITION
Maximum 1.0 ppm. 5-[ (2RS)-3-[(I, I-Dimethylethyl)amino]-2-hydroxypropoxy]-
Loss on drying (2.2.32) 3,4-dihydroquinolin-2(1H)-one hydrochloride.
Maximum 12.0 per cent, determined on 1.000 g by drying in Content
an oven at 105 °C. 99.0 per cent to 101.0 per cent (dried substance).
Tora! ash (2.4.16) CHARACTERS
Maximum 40.0 per cent. Appearance
White or almost white crystals or crystalline powder.
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2022 CarteoloI Hydrochloride 1-449
Solubility - unspecified impurities: for each impurity, not more than the
Soluble in water, sparingly soluble in methanol, slightly area of the principal peak in the chromatogram obtained
soluble in ethanol 96 per cent, practically insoluble in with reference solution (b) (0. IO per cent);
methylene chloride. - total: not more than half the area of the principal peak in
IDENTIFICATION
(0.5 per cent);
Comparison Ph. Bur. reference spectrum of camolol the chromatogram obtained with reference solution (b)
hydnxhloride. (0.02 per cent).
B. It gives reaction (a) of chlorides (2.3.1). Loss on drying (2.2.32)
TESTS Maximum 0.5 per cent, determined on 1.000 g by drying in
Appearance of solution an oven at 105°C for 3 h.
The solution is clear (2.2.1) and colourless (2.2.2, Sulfated ash (2.4.14)
Method II). Maximwn 0.1 per cent, determined on 1.0 g.
Dissolve 0.300 g in waleI'R and dilute to 10 mL with the
ASSAY
same solvent.
Dissolve 0.250 g in 60 mL of ethanol (96 per cent) R.
pH (2.2.3) Add 5.0 mL of 0.01 M i1ydrochloric acid. Carry out a
5.0 to 6.0. potentiometric titration (2.2.20), using 0.1 A-I sodium
Dissolve 0.250 g in carbon dioxide-free waterR and dilute to hydroxide. Read the volume added between the 2 points of
25 mL with the same solvent. inflexion.
Related substances 1 mL of 0.1 M sodium hydroxide is equivalent to 32.88 mg of
Liquid chromatography (2.2.29). C16H25N203CI.
Testsolution Dissolve 20.0 mg of the substance to be STORAGE
examined in the mobile phase and dilute to 10.0 mL with In an airtight container.
the mobile phase.
IMPURITIES
Reference solution (aJ Dilute 1.0 mL of the test solution to Specified impurities H.
Other detectable impuruies (the following substances would, if
Reference solution (b) Dilute 1.0 mL of reference solution (a) present at a sufficient level) bedetected by one or otherof the tests
to 10.0 mL with the mobile phase. in the monograph. They are limited by the general acceptance
Reference solution (c) Dissolve 10 mg of carnolol for system criterion for other/unspecified impun'ties and/or by the general
suitaln1ilY CRS in the mobile phase and dilute to 5 mL with monograph Substances for phannaceuuCaI use (2034). It is
the mobile phase. therefore not necessary to identifythese impurities for
Reference solution (d) Dilute 5.0 mL of reference demonstration ofcompliance. Sa also 5.10. Control of impurities
solution (b) to 10.0 mL with the mobile phase. in subsronces for pharmaceutical use) A, B) C, D, E, F) G, 1.
Column:
=
- size: I:::: 0.25 m, 0 4.6 mm;
- stationary phase: octade<:y/si/yl silica gelfor chromatography R
(5 urn).
Mobile phase Mix I volume of methanol R2, 20 volumes of
acetonitrile R and 79 volumes of a 2.82 gIL solution of sodium
hexanesulfonate R. A. 4,6,7 ,8-tetrahydroquinoline-2,5(IH,3H)-dione,
Flow rau 1 mUmin.
Injection 20~.
onU ,OH
with camololfor $)'Sum suitability CRS to identify the peak

due to impurity H. B. 5-hydroxy-3,4-dihydroquinolin-2(1H)-one,
System suitabr1ity:
o~
- the chromatogram obtained with reference solution (c) is
similar to the chromatogram provided with carteolof for HN::f? H
0 <,><.] ° and enantiomer
system suitabilily CRS; the peaks due to impurity H and
carteolot show base-line separation; ""I
- signal-co-noise ratio: minimum 10 for the principal peak in
the chromatogram obtained with reference solution (d); C.5-[[(2RS}-oxiran-2-yl]methoxy]-3,4-dihydroquinolin-2
- numberof theoretical plates: minimwn 6000, calculated for (IH)-one,
o~ 0~C1
reference solution (a). H OH
Limits: HN::f?
- impun'ty H: not more than twice the area of the principal and enanliomer
peak in the chromatogram obtained with reference ""I
solution (b) (0.2 per cent); .
D.5-[(2RS}-3-chloro-2-hydroxypropoxy]-3,4-
dihydroquinolin-2(1H)-one,
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1-450 Carvedilol 2022
Content
CHARACTERS
Appearance
E. 5,5'- [(2-hydroxypropan-I,3-diyl) bis(oxy)]bis(3,4-
dihydroquinolin-2(1H)-one), Solubility
Practically insoluble in water, sparingly soluble in methylene
O~O~OCH,
chloride, slightly soluble in ethanol (96 per cent). It is
H OH
practically insoluble in dilute acids.
HN: andenanlioma<
IDENTIFICATION
F. 5-[(2RS)-2-hydroxy-3-methoxypropoxy]-3,4-
Comparison caroedilol CRS.
dihydroquinolin-2(1H)-one,
If the spectra obtainedshow differences, dissolve the
0:CO
HN #'"
,%1
O~OH
H OH
andenantiomer
separately in 2-propanol R, evaporate [Q dryness and record
TESTS
Related substances
G. 5-[(2RS)-2,3-dihydroxypropoxy]-3,4-dihydroquinolin-2 Liquid chromatography (2.2.29).
(1H)-one, Test soluticn Dissolve 25 mg of the substance to be
the mobile phase.
H. 5-[ (2RS)-3-[ (I, l-dimethylethyl)amino]-2- Reference solulion (b) Dissolve 5 mg of caroedilo/
hydroxypropoxy]quinolin-2(1H)-one, impurity C CRS in 5.0 mL of the mobile phase and dilute to
solution to 100.0 mL with the mobile phase. Dilute 1.0 mL
of this solution to 10.0 mL with the mobile phase.
Reference solution (c) Dissolve 5 mg of caroedUol for system
and enanliomer
suitability CRS (containing impurities A and D) in the mobile
Column:
I. 7-bromo-5-[(2RS)-3-[(I,I-(dimethylethyl)amino]-2-
- size: 1= 0.150 m, 0 ;:;; 4.6 mm;
hydroxypropoxy]-3,4-dihydroquinolin-2(1H)-one.
- stationary phase: end-<:apped oay/silylsilica gelfor
_____________ ~ Ph,,, chromatography R (5 um),
Mobilephase Dissolve 1.77 g ofporassium dihydrogen
phosphate R in water R and dilute to 650 mL with the same
solvent; adjust to pH 2.0 with phosphoric acid R and add
Carvedilol 350 mL of acetonitrile R.
(Ph. Eur. monograph 1745) Fluco rate 1.0 mUmin.
Injection 20 ~L.
Run time 6 times the retention time of carvedilol.
Identification of impurities . Use the chromatogram supplied
with caroedilol for system suitability CRS and the
the peaks due to impurities A and D; use the chromatogram
406.5 72956-09-3 obtained with reference solution (b) to identify the peak due
to impurity C.
Action and use ReJauve retention With reference to carvedilol (retention
Beta-adrenoceptor antagonist; arteriolar vasodilator. time = about 4 min): impurity A = about 0.5;
Preparation impurity C =abeut 2.9; impurity D =about 3.8.
Carvedilol Tablets System suitability:
Ph'" _ - resolution: minimum 3.5 between the peaksdue to
impurity A and cervedilol in the chromatogram obtained
DEFINITION with reference solution (c);
(2RS)-I-(9H-Carbazol-4-yloxy)-3-[[ 2-(2-
methoxyphenoxy)ethyl]amino]propan-2-01.
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2022 Castor Oil 1-451
~ signal-to-noise ratio: minimum 10 for the peak due to

impurity C in the chromatogram obtained with reference OH
f '"
~
solution (b). l... ~O NH

Limits: OH ('" I ~
I 0
peak area of impurity A by 2.0;
- impun"ty A: not more than twice me area of the principal
..? ~OCH,
HN U
- impurity D: not more man 1.5 times Ute area of the
principal peak in the chromatogram obtained with B. I, I '-[[2-(2-methoxyphenoxy)ethyl)nilrilo)bis[3-(9H-
reference solution (a) (0.15 per cent); carbazol-I-yloxyjpropan-z-ol],
- impun""Y C: not more than the area of the corresponding
areaof the principal peak in the chromatogram obtained and enanliomer
with reference solution (a) (0.10 per cent),
- sum of impurities other than C: not more than 5 times the
C. (2RS)-I-[benzyl[2-(2-methoxyphenoxy)ethyl]antino)-3-
(9H-carbazol-4-yloxy)propan-2-0I,
(0.05 per cent).
Maximurn 0.5 per cent) determined on 1.000 g by drying in
an oven at 105 "C.
Maximum 0.1 per cent) determined on J.O g.
ASSAY
Dissolve 0.350 g in 60 ffiL of anhydrous auric acidR. Titrate
with 0.1 M perchloric acid, determining the end-point
I mL of 0.1 M perchloric acid is equivalent to 40.65 mg of D. 1-(9H-carbazol-4-yloxy)-3-[4- [2-hydroxy-3- [[2-(2-
C,.,I!,oN,O-r- methoxyphenoxy)ethyl]antino)propoxy)-9H-carbazol-9-yl)
propan-2-ol.
IMPURITIES _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Spedfied ;mpun"ties A J C, D.
Otherdetectable impurities (thefollowing subsronces would, if
in the monograph. They are limited by the general a«eptance
cviterion for other/unspedfied impU1;ties and/or by the general Hydrogenated Castor Oil
monograph Substances for pharmaceurical use (2034). It is
demonstration of compliance. See also 5.10. Control of impuniies Action and use
in substances for pharmaceutical use) B. Excipient.
PhE<r _
DEFINITION
Fatty oil obtained by hydrogenation of Virgin costor oil (0051)
or Refined COStar oil (2367) or a mixture of both. It consists
mainly of the triglyceride of 12-hydroxystearic «123)-12-
hydroxyoetadecanoic) add.
CHARACTERS
Appearance
Fine) almost white or pale yellow powder or almost white or
A. 1-[[9-[2-hydroxy-3-[[2-(2-methoxyphenoxy)ethyl]
pale yellow masses or flakes.
antino]propyl] -9H-carbazol-4-yl] oxy]-3- [[2-(2-
methoxyphenoxy)ethyl]antinojpropan-2-01, Solubility
Practically insoluble in water, slightly soluble in methylene
chloride, veryslightly soluble in anhydrous ethanol,
practically insoluble in light petroleum.
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1-452 Castor Oil 2022
IDENTIFICATION
A. Melting point (2.2.N): 83 °C to 88 °C. area of the peaks due to any specified or unspecified fatty acid
B. Hydroxyl value (see Tests). methyl esters;
C. Composition of fatty acids (see Tests).

TESTS R.: relative correction factor for the peak due to methyl
12-hYdroX)'$teaca te:
Acid value (2.5.1)
Maximum 4.0, determined on 10.0 g dissolved in 75 mL of
hot emanal (96 perunO R.
145 to 165, determined on a warm solution. R.: I for peaks corresponding to each of the other specified fatty
Iodine value (2.5.4, Memod A) adds or any unspecified fatty acid;
ml.. mass ofmelhyl12·hydroxySleacate in the reference solution;
Maximum 5.0. m2,.. mass of methyl stearate in the reference solution;
Alkaline impurities AI.. area of the peak due (0 methyl l2-hydroxystearate in the
Dissolve 1.0 g with gentle heating In a mixture of 1.5 mL of chromatogram obtained with the reference SOlution;
A 2.r area of the peak due to methyl stearate in the chromatogram
ethanol (96 per unO Rand 3 mL of ",Inene R. Add 0.05 mL obtained with ihe reference solution.
of a 0.4 gIL solution of bromophenol blue R in emanol
(96 per eenO R. Not more than 0.2 mL of 0.01 M hydrochloric
acid is required to change the colour of me indicator to Composition of the fatty-acidfraction of me oil:
yellow. - palmitic acid: maximum 2.0 per cent;
Composition of fatty acids - steam add: 7.0 per cent to 14.0 per cent;
Gas chromatography (2.4.22, Method A) with the following - arachidic acid: maximum 1.0 per cent;
modifications. Use the mixture of calibrating substances in - tz-oxosuosic acid: maximum 5.0 per cent;
Table 2.4.22.-3. - 12-hydroxystean"c acid: 78.0 per cent to 91.0 per cent;
- atry' otherfatty acid: for each fatty acid, maximum
Test solution Introduce 75 rng of the substance to be
3.0 per cent.
examined into a 10 mL centrifuge tube with a screw cap.
Dissolve in 2 mL of 1,I-dimethy/ethyl methyl ether Rl by
shaking and heat gently (50-60 0C). Add, when still warm, STORAGE
1 mL of a 12 gIL solution of sodium R in anhydrous In a well-filled container.
methanol R J prepared with the necessary precautions, and mix
vigorously for at least 5 min. Add 5 mL of distiUed wow R IMPURITIES
and mix vigorously for about 30 s. Centrifuge for 15 min at
1500 g. Use the upper layer.
o
Reference soluticn Dissolve 50 mg of methyl
12-hydroxysuara,. CRS and 50 mg of methylsuara,. CRS in A. 12-oxooetadecanoic acid (12-oxostearic acid).
10.0 mL of 1,I-dimethylethyl methyl ether Rl. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Phf"
Column:
- size: / == 30 m; 0 == 0.25 rnm;
- stationary phase: macragol20 000 R (film thickness Polyoxyl Castor Oil
0.25 pm).
Cartier gas helium for chromatography R. (MacragolglYcerol Rit;inalea,., Ph. Eur. monograph
Flow ra" 0.9 mUmin. 1082)
Split rat;" 1:100. Action and use
Temperature: Excipient.
Phf" _
Time Temperoture
(min) Cq DEFINITION
Column 0-55 215 Contains mainly ricinoleyl glycerol ethoxylated with
Injection port 250 30-50 molecules of ethylene oxide (nominal value), with
Detector '-; 250 small amounts of macrogol ricinoleate and of me
corresponding free glycols. It results from the reaction of
Detection Flame ionisation. castor oil with ethylene oxide.
Inj«ticn I ~L. CHARACTERS
Sysum suitabJity: Appearance
- symmetry factor: 0.7 to 1.5 for the peak due to methyl Clear, yellow viscous liquid or semi-solid.
stearate in the chromatogram obtained with the test Solubility
solution. Freely soluble in water, very soluble in methylene chloride,
Calculate the fraction of each fatty acid using the following freely soluble in ethanol (96 per cent).
expression: Relative density
About 1.05.
A_'l:,s,c/LA.'l:,.J,c x 100 per cent m/m
VIscosity
AJ",~ corrected peak area of the fatty acid in the lest solution: 500 ml'a-s to 800 rnl'a-s at 25 °C.
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IDENTIFICATION Table 1082.-1

A. Iodine value (see Tests). Ethylene oxide units per Hydroxyl value Saponification value
B. Saponification value (see Tests). molecule (nominal value)
C. Thin-layer chromatography (2.2.27). 30 - 35 65 - 82 60 - 75

50 48 -68 38 - 52
Testsolution To 1 g of the substance to be examined add
100 mL of a 100 gIL solution of potassium hydroxide Rand
boil undera reflux condenser for 30 min. Allow to cool. Residual ethylene oxide and dioxan (2.4.25)
Acidify the solution with 20 mL of hydrochloric acid R. Shake Maximum I ppm of residual ethylene oxide and 10 ppm of
the mixture with 50 mL of ether R and allow to stand until residual dioxan.
separation of the layers is obtained, Transfer the dear upper Water (2.5.12)
layerto a suitable tube, add 5 g of anhydrous sodium sulfate R, Maximum 3.0 per cent, determined on 2.000 g.
close the tube and allow to stand for 30 min. Filterand Total ash (2.4.16)
evaporate the filtrate to dryness on a water-bath. Dissolve Maximum 0.3 per cent, determined on 2.0 g.
50 mg of the residue in 25 mL of ether R.
Reference solution Dissolve 50 mg of ricinoleic acid R in STORAGE
methylene chloride Rand dilute (Q 25 mL with the same Protected from light.
solvent. LABELLING
Plate TLC ocrade<y/silyl silica gel plate R. The label srates:
lvlobile phase methylene chloride R, gladal acetic addR, - the amountof ethylene oxide reactedwith castor oil
ace"'ne R (10:40:50 VIVIV). (nominal value),
Applicarion 2 ~L.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Drying In a current of cold air.
Detection Spray with an 80 gIL solution of phosphomolybdi<:
acidR in 2-proponol R and heat at 120 °C for 1-2 min.
Results The principal spot in the chromatogram obtained Hydrogenatea Polyoxyl Castor Oil
with the test solution is similar in position and colour to the
principal spot in the chromatogram obtainedwith the (Macrogolglycerol Hydroxystearate, Ph. Eur.
reference solution. monograph 1083)
D. Place about 2 g of the substance to be examined in a test- PhE" _
tube and add 0.2 mL of sulfuric acidR. Close the tube using
a stopper fitted with a glass tube bent twice at right angles. DEFINITION
Heat the tube until white fumes appear. Collect the fumes in Contains mainly rris(12-hydroxystearyl) glycerol ethoxylated
1 mL of mercutic chloride solution R. A white precipitate is with 7 to 60 molecules of ethyleneoxide (nominalvalue),
formed and the fumes tum a filterpaper impregnated with with small amounts of macrogol hydroxystearate and of the
alkaline potassium tetraiodomercunue solution R black. corresponding freeglycols. It results from the reaction of
hydrogenated castor oil with ethylene oxide.
TESTS
Solution S CHARACTERS
Dissolve 5.0 g in carbon dioxide-free water R and dilute to Appearance
50 mL with the sante solvent. - if less than 10 units of ethylene oxide per molecule:
yellowish, turbid, viscous liquid;
- if more than 20 units of ethylene oxide per molecule:
Solution S is not more opalescent than reference
white or yellowish semi-liquid or pastymass.
suspension III (2.2.1) and not more intensely coloured than
reference solution BY5 (2.2.2, Method 11). H intended for use Solubility
in the manufacture of parenteral preparations, solution S is - if less than 10 units of ethylene oxide per molecule:
not more intensely coloured than reference solution BY6 practically insoluble in water, soluble in acetone,
(2.2.2, Merhad II). dispersible in ethanol(96 per cent);
- if more than 20 units of ethylene oxide per molecule:
Alkalinity
freely solublein water, in acetone and in ethanol
Dissolve 2.0 g in a hot mixture of 10 mL of water Rand
(96 per cent), practically insoluble in light petroleum.
10 mL of ethanol (96 per cent) R. Add 0.1 mL of bromothymol
blu.e solution RI. Not more than 0.5 mL of 0.1 M hydrochloric IDENTIFICATION
acid is required to change the colourof the indicator to A. Iodine value (see Tests).
yellow. B. Saponification value (see Tests).
Acid value (2.5.1) C. Thin-layer chromatography (2.2.27).
Maximum 2.0, determined on 5.0 g. Test solution To 1 g of the substance to be examined, add
Hydroxyl value (2.5.3, MethodA) 100 mL of a 100 gIL solution of potassium hydroxide Rand
See Tahle 1082.-1. boil undera reflux condenser for 30 min. Allow to cool.
Iodine value (2.5.4) Acidify the solution with 20 mL of hydrochlori< acid R. Shake
25 to 35. the mixture with 50 rnL of ether R and allow to stand until
separation of the layers is obtained. Transfer the clear upper
Saponification value (2.5.6) layer to a suitable tube, add 5 g of anhydrous sodium suifate R,
See Tahle 1082.-1. close the tube and allow to stand for 30 min. Filter and
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1-454 Castor Oil 2022
evaporate the filtrate to dryness on a water-hath. Dissolve Water (2.5.12)

50 mg of the residue in 25 mL of ether R. Maximum 3.0 per cent, determined on 2.000 g.
Re/ereJlu solution Dissolve 50 rng of 12-hydroxystean"c acidR Total ash (2.4. 16)
in methylene chloride R and dilute to 25 mL with the same Maximum 0.3 per cent, determined on 2.0 g.
solvent.
LABELLING
Piat« TLC /Xtadecylsilyl silica gelplate R. The label states the number of ethylene oxide units per
Mobile phase methylene chloride R, glacial acetic add R, molecule (nominal value).
"",tone R (10:40:50 VIVIJI). ______________ ~ Pr>E"
ApplicaliJJn 2 ~L
Detection Spray with a 80 gIL solution of phosphomolybdic Refined Castor Oil
acidR In. 2-propanol R and heat at 120°C for about 1-2 min.
Results The principal spot in me chromatogram obtained
with the test solution is similar in position and colour to the Pr>E" ~ _
principal spot in the chromatogram obtained with me DEFINITION
referencesolution. Fatty oil obtained from the seeds of Ricinus communis L.
D. Place about 2 g in a test-tube and add 0.2 mL of sulfuric by cold expression. It is then refined. A suitable antloxidanr
acid R. Close the tube using a stopper fitted with a glass tube may be added.
bent twice at right angles. Heat the tube until white fumes
appear. Collect the fumes in 1 mL of mercuric chloride PRODUCTION
solution R. A white precipitate is formed and the fwnes turn a During the expression step, the temperature of the oil must
filter paper impregnated with alkalinepotassium not exceed 50 -c,
letraiodomen:urate solution R black. CHARACTERS
TESTS Appearance
Solution S Clear, almost colourless or slightly yellow, viscous,
Dissolve 5.0 g of mac rogol glycerol hydroxystearate with less hygroscopic liquid.
than 40 units of ethylene oxide per molecule in a mixture of Solubility
50 volumes of acetone Rand 50 volumes of anhydrous Slightly soluble in light petroleum, miscible with ethanol
ethanol R and dilute to 50 mL with the same mixture of (96 per cent) and with glacial acetic acid.
solvents. Relative density
Dissolve 5.0 g of macrogolgtycerol hydroxystearate with About 0.958.
40 units or more of ethylene oxide per molecule in carbon Refractive index
dioxide-free wol<r R and dilute to 50 mL with the same About 1.479.
solvent.
Viscosity
Appearance of solution About 1000 ml'a-s.
Solution S is not mo.re opalescent than reference
suspension ill (2.2.1) and not more intensely coloured than IDENTIFICATION
reference solution BY. (2.2.2, MelhodII). First identificatUm: B, C.
Alkalinity Second identification: A, B.
To 2 mL of solution S add 0.5 mL of bromothymol blue A. A mixture of 2 mL of the substance to be examined and
solution Rl. The solution is not blue. 8 mL of ethanol (96 per cen!J R is clear (2.2. 1).
Acid value (2.5.1) B. Specific absorbance (see Tests).
Maximum 2.0, determined on 5.0 g. C. Composition of fatty acids (see Tests).
Hydroxyl value (2.5.3, MethodA) TESTS
See Table 1083.-1. Appearance '
Iodine value (2.5.1) The substance to be examined is clear (2.2.1) and not more
Maximum 5.0. intensely coloured than reference solution BY3 or Y 3 (2.2. 2.J
Saponification value (2.5.6) .,Method/).
See Table 1083.-1. Optical rotation (2.2.7)
+ 3.5"to + 6.0°.
Table 1083.-1
Specific absorbance (2.2.25)
Ethylene oxide units per Hydroxyl value Saponifica.tion value 0.7 to 1.5, determined at the absorption maximum at
molecule (nominal value)
270 run.
7 115 - 135 125 - 140
To 1.00 g add ethanol (96 per CetI!J R and dilute to 100.0 mL
25 70 - 90 70 - 90
40 57 - 80 45·69
60 45 - 67 40 - 51 Acid value (2.5.1)
Maximum 0.8.
Residual ethylene oxide and dloxan (2.4.25) Dissolve 5.00 g in 25 mL of the prescribed mixture of
Maximum I ppm of residual ethylene oxide and 10 ppm of solvents.
residual dioxan.
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Hydroxyl value (2.5.3, MethodA) AI area of the peak due to methyl ricinoleate in me chromatogram
obtained wilh the reference solution;
Minimum 160.
A2 area of the peak due to methyl stearate in the chromatogram
Peroxide value (2.5.5, MethodA) obtained with the reference solution.
Maximum 5.0.
Unsaponifiable matter (2.5.7) Composition 01 tilelatty-acid/raetion 01 the oil:
- palmitic acid: maximum 2.0 per cent;
- stearic acid: maximum 2.5 per cent;
Oil obtained by extraction and adulteration - oleic acid and isomer: 2.5 per cent [0 6.0 per cent;
In a ground-glass-stoppered tube about J 25 mm long and - finoleic add: 2.5 per cent to 7.0 per cent;
18 mm in internal diameter, thoroughly mix 3 mL of the - linolenic acid: maximum 1.0 per cent;
substance to be examined with 3 mL of carbon disulfide R. - eicosenoic acid: maximum 1.0 per cent;
Shake for 3 min with 1 mL of sulfuric acidR. The mixture is - ricinoleic acid: 85.0 per cent to 92.0 per cent;
less intensely coloured than a freshly prepared mixture of - any otherfatty acid: for each fany acid, maximwn
3.2 mL oflerri< chloride solution RI, 2.3 mL of waterRand 1.0 per cent.
0.5 mL of dIlute ammonia RI.
Water (2.5.32)
ComposItion of fatty acids Maximwn 0.3 per cent, or maximum 0.2 per cent If intended
Gas chromatography (2.4.22) with the following for use in the manufacture of parenteral preparations)
modifications. determined on 1.00 g.
STORAGE
Test solution Introduce 75 mg of the substance to be In an airtight, well-tilled container, protected from light.
examined into a 10 mL centrifuge tube with a screw cap. If intended for use in the manufacture of parenteral
Dissolve in 2 mL of 1,I-<1imethylethyl methylether RI with preparations, store under an inert gas.
shaking and heat gently (50-60 "C). Add, while still warm,
I mL of a 12 f!!L solution of sodium R in anhydrous LABELLING
methanol R, prepared with the necessaryprecautions, and The label states, where applicable, that the substance is
shake vigorously for at least 5 min. Add 5 mL of distilled suitable for use in the manufacture of parenteral preparations
water R and shake vigorously for about 30 s. Centrifuge for and the name of the men gas.
15 min at 1500 g. Use the upper layer. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE«
Reference solution Dissolve 50 mg of methylricinoleate CRS

and 50 mg of methylstearate CRS in 10.0 mL of
1,I-<1imethylethyl methyletherRI.
Virgin Castor Oil ***
Column:
- material: fused silica; *** ***
- size: 1= 30 m, 0 = 0.25 mm; (Ph. Eur. monograph 0051) ***
- stationary phase: macrogol 20 000 R (film thickness Action and use
0.25 urn). Stimulant laxative; emollient.
Cartier gas helium for chromatography R. Preparation
Flow rate 0.9 mllmin. Zinc and Castor Oil Ointment
Splli ratio 1:100. PhE" _
Temperature:
DEFINITION
Tim. Temperature Fatty oil obtained by cold expression from the seeds of
(min) ('C) Ricinus communis L. A suitable antioxidant may be added.
Column 0-55 215 PRODUCTION
Injection port 25. During the expression step, the temperature of the oil must
Detector 25. not exceed 50°C.
CHARACTERS
Appearance
Injection 1 IlL. Clear at 40 °C, slightly yellow, viscous, hygroscopic liquid.
System suitability: Solubility
- symmetry factor: 0.7 to 1.5 for the peak due to methyl Slightly soluble in light petroleum, miscible with ethanol
stearate in the chromatogram obtained with the test
(96 per cent) and with glacial acetic acid.
solution.
Relative density
Calculate the percentage content of each fatty acid by the
About 0.958.
normalisation procedure.
Correct the area of the peak due to methyl ricinoleate, by
Refractive index
About 1.479.
multiplying by a factor R calculated using the following
expression: IDENTIFICATION
First identification: B) C.
A. A mixture of 2 mL of the substance to be examined and
mass of methyl ricinoleate in me reference solution; 8 mL of ethand (96 percent) R is deas (2.2./).
mass of melhyl stearate in the reference solution; B. Specific absorbance (see Tests).
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1-456 Cefaclor 2022
C. Composition of fatty acids (see Tests). expression:

TESTS
+ 3.5° to + 6.0 0 •
Specific absorbance (2.2.25) mass of melhyl ricinoleate in the reference solution;
Maximum 0.7, determined at the absorption maximum at massor methyl stearate in the reference solution;
area of the peak due to melhyl ricinoleate in the chromatogram
270 nm. obtained with the reference solution.
To 1.00 g add ethanol (96 per cent) R and dilute to 100.0 mL A, area of the peak due to methyl stearate in the chromatogram
with the same solvent. obtained with the reference solulion.
Acid value (2.5.1) Composition of thefaUy-acid fraction of the oil:

Maximum 1.5. - palmitic add: maximum 2.0 per cent;
Dissolve 5.00 g in 25 mL of the prescribed mixture of - stearic acid: maximum 2.5 per cent;
solvents. - oleic add and isomer: 2.5 per cent to 6.0 per cent;
Hydroxyl value (2.5.3, Me'/wd A) - linoleic acid: 2.5 per cent to 7.0 per cent;
Minimum 160. - linolenic acid: maximum 1.0 per cent;
Peroxide value (2.5.5, Me/hod A)
- eicosenoic acid : maximum 1.0 per cent;
- ricinoleic acid: 85.0 per cent to 92.0 per cent;
Maximum 10.0.
- atryother jatty acid: for each fatty acid, maximum
Unsaponifiable matter (2.5.7) 1.0 per cent.
Water (2.5.32)
Composition of fatty acids Maximum 0.3 per cent, determined on 1.00 g.
Gas chromatography (2.4.22) with the following
modifications. STORAGE
In an airtight, well-filled container, protected from light.
_____________________ "'E"
Test solmion Introduce 75 mg of the substance to be
examined into a 10 mL centrifuge tube with a screw cap.
Dissolve in 2 mL of 1,I-dimethy/ethyl methylether Rl with
shaking and heat gently (50-60 "C). Add, while still warm,
I mL of a 12 gIL solution of sodium R in auhydrous Cefaclor
methanol R, prepared with the necessary precautions, and mix
vigorously for at least 5 min. Add 5 mL of distilled water R (ph. Bur. monograph 0986)
and mix vigorously for about 30 s. Centrifuge for 15 min at
1500 g. Use the upper layer.
Reference solUlwn Dissolve 50 mg of methyl ricinoleate CRS
H,O
and 50 rng of methylstearate CRS in 10.0 mL of
1,I-;Jimethylethyl methylether Rl.
Column:
- size: 1= 30 m, 0 = 0.25 mm; 385.8 70356-03-5
- stationary phase: mocrogol 20 000 R (film thickness
0.25 pm). Action and use
Cephalosporin antibacterial.
Flow rale 0.9 mllmin. Preparations
Cefaclor Capsules
Split ratio 1:100.
Cefaclor Oral Suspension
Temperature:
Cefaclor Prolonged-release Tablets
Time Temperature "'E" _
(min) CC)
Column 0-55 215 DEFINITION
Injection port 2'0 (6R, 7R)-7 -[[(2R)-2-Amino-2-phenylacetyl]amino]-3-cWoro-
Detector 2'0 8-oxo-5-thia-I-azabicyclo[4.2.0] ocr-z-ene-2-carboxylie acid
monohydrate.
Detection Flame ionisation. Semi-synthetic product derived from a fermentation product.
Injection 1 ~L. Content
System suitability: 96.0 per cent to 102.0 per cent of CI5Hl4CIN304S
- symmetry fatter. 0.7 to 1.5 for the peak due to methyl (anhydrous substance).
stearate in the chromatogram obtained with the test CHARACTERS
solution. Appearance
Calculate the percentage content of each fatty acid by the White or slightly yellow powder.
normalisation procedure. Solubility
Correct the area of the peak due to methyl ricinoleate, by Slightly soluble in water, practically insoluble in methanol
multiplying by a facror R calculated using the following and in methylene chloride.
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2022 Cefaclor 1-457
IDENTIFICATION Water (2.5.12)

Infrared absorption spectrophotometry (2.2.24). 3.0 per cent to 6.5 per cent, determined on 0.200 g.
Comparison cefador CRS. ASSAY
TESTS Liquid chromatography (2.2.29).
pH (2.2.3) Test solution Dissolve 15.0 mg of the substance to be
3.0 to 4.5. examined in the mobile phase and dilute to 50.0 mL with
Suspend 0.250 g in carbon dioxide-free waterR and dilute to the mobilephase.
10 mL with the same solvent. Reference solutien (a) Dissolve 15.0 mg of cefador CRS in
Specific optical rotatIon (2.2.7) the mobile phase and dilute to 50.0 mL with the mobile
+ 101 to + III (anhydrous substance). phase.
Dissolve 0.250 g in a 10 gIL solution of hydrochloric acid R Reference sdution (b) Dissolve 3.0 mg of cefador CRS and
and dilute to 25.0 mL with the same solution. 3.0 mg of ddta-l-cefador CRS (impurity D) in the mobile
Related substances
Column:
- size: 1= 0.25 m, 0 = 406 mm;
Test solution Dissolve 50.0 mg of the substance to be - statiouary phase: oCUldecylsuyl silica gelfor chromatography R
examined in 10.0 mL of a 2.7 gIL solution of sodium (5 urn).
dihydrogen phosphate R adjusted [0 pH 2.5 with phosphoric
Mobile phase Add 220 mL of methanol R to a mixture of
acid R.
780 mL of water R, 10 mL of rnOethylamine Rand 1 g of
Reference solution (a) Dissolve 2.5 mg of cefador CRS and sodium penumesulfonate R, then adjust to pH 2.5 with
5.0 mg of deua-l-afador CRS (impurity D) in 100.0 mL of a phosphoric acidR.
2.7 gIL solution of sodium dihydrogen phosphate R adjusted to
pH 2.5 with phosphoric acid R.
Detecdon Spectrophotometer at 265 nm.
100.0 mL with a 2.7 gIL solution of sodium dihydrogen Injeetiou 20~.
phosphate R adjusted to pH 2.5 with phosphoric acidR. Systemsuitability:
Column: - resolution: minimum 2.5 between the peaks due to cefaclor
- size: 1= 0.25 m, 0 = 4.6 mrn; and impurity D in the chromatogram obtained with
- stationary phase: end-capped oCUldeeylsj(y1 suica gelfor reference solution (b); if necessary, adjust the
chromatography R (5 .um). concentration of methanol in the mobile phase;
- symmetry factor. maximum 1.5 for the peakdue to cefaclor
Mobuephase:
in the chromatogram obtained with reference solution (b)j
- mobile phase A: 7.8 gIL solution of sodium dihydrogen
phosphate R adjusted to pH 4.0 with phosphori< acidR;
1.0 per cent after 6 injections of reference solurion (a).
- mobile phaseB: mix 450 mL of acetonitrile R with 550 mL
of mobile phase Aj IMPURITffiS

(mln) (per cent YIl1 (per cent JIM
0·30 95 -+ 75 5 ---> 25
30 - 45 75 --> 0 25 -+ 100
45 - 55 o 100
A. (2R)-2-amino-2-phenylacetic acid (phenylglycine),
Injection 20~.
- resolution: minimum 2 between the peaks due to cefaclor
and impurity D; if necessary, adjust the acetonitrile
B. (6R,7R)-7-antino-3-chloro-8-oxo-5-thia-l-
content in the mobile phase;
azabicydo[4.2.0)oct-2-ene-2-carboxylic acid,
- symmetry factor: maximum 1.2 for the peakdue to
cefac1or; if necessary, adjust the acetonitrile contentin the
mobile phase.
Limits: H.NH2~)-~:J
o J::CI
- any impun"ty: for each impurity, not more than 0.5 times
the area of the principal peakin the chromatogram
ctr
"" I
% 0
-t--}. H H
S
C. (6R,7R)-7 -[[(2S)-2-amino-2-phenylacetyl)amino]-3-
(2 per cent); chlcro-s-oxo-s-thia-I-azabicydo[4.2.0) oct-2-ene-2-
- disregard limit: 0.1 times the area of the principal peak in carboxylic acid,
(0.1 per cent).
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1-458 Cefadroxil Monohydrate 2022
PhE" _
DEFINITION
(6R,7R)-7-[[(2R)-2-Amino-2-( 4-hydroxyphenyl)acetyl]
aminol-3-methyl-8-oxo-5-thia-l-azabicyclo[4.2.OJ oct-2-ene- 2-
carboxylic acid monohydrate.
D. (2R,6R,7R)- and (2S,6R,7R)-7-[[(2R)-2-amino-2- Semi-synthetic productderived from a fermentation product.
phenylaceryljamino]-3-chloro-8-oxo-5-thia- I-azabicyclo Content
[4.2.0]oct-3-ene-2-carboxylic acid (delta-3-cefaclor), 95.0 per cent to 102.0 per cent (anhydrous substance).
o
CHARACTERS
~
CI
HNH, HN I Appearance
~~0 White or almost white powder.
U ~ CO"'S
Solubility
Slightly soluble in water, very slightly soluble in ethanol
(96 per cent).
E. 2-[[(2R)-2-amino-2-phenylacetyljamino]-2-(5-chloro-4-
oxo-3,4-dihydro-2H-l,3-rhiazin-2-yl)acetic acid, IDENTIFICATION
(XON
OH
,p-
'"
Comparison
TESTS
pH (2.2.3)
4.0 to 6.0.
cejadroxiJ CRS.
F. 3-phenylpyrazin-2-01,
'
·
Suspend 1.0 g in carbon dioxide-free water R and dilute to
o H~.co,H
orr
• CH2
~'#s
H NH:! N Specific optical rotatIon (2.2.7)
ondeplme,,'C' + 165 to + 178 (arthydrous substance).
I H H Dissolve 0.500 g in waterR and dilute to 50.0 mL with the
'" 0 same solvent.
G. (2R,6R,7R)- and (2S,6R,7R)-7-[[(2R)-2-amino-2- Related substances
phenylacetyl]amino]-3-methylene-8-oxo-5-thia-l-
azabicyclo[4.2.0joetane-2-earboxylic acid (isocefalexine), Test solution Dissolve 50.0 mg of the substance to be
mobile phase A.
o:J:
H" NH, 0 yeo'H
o CI Reference solution (a) Dissolve 10.0 mg of IHI.-(4-
~ H. NH )---~ .... ~ hydroxyphenyl)glycine CRS (impurity A) in mobile phase A
~~tts and dilute to 10.0 mL with mobile phase A.
V~ Reference solution (b) Dissolve 10.0 mg of

r-ammodesacetoxycephalosporanic acid CRS (impurity B) in
phosphare buffer solution pH 7.0 R5 and dilute to 10.0 mL
H. (6R, 7R)-7-[[(2R)-2-[[(2R)-2-amino-2-phenylacetyl]
with the same buffer solution.
amino J-2-phenylacetyl]amino ]-3-chloro-8-oxo-5 -thia-f-
azabicyclo[4.2.0joet-2-ene-2-earboxylic acid (N- Reference solmion (c) Dilute 1.0 mL of reference solution (a)
phenylglycyl cefaclor). and 1.0 mL of reference solution (b) to 100.0 mL with
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" mobile phase A.
Reference solution (d) Dissolve 10 mg of dimethylformamilk R
and 10 mg of dimethylacetamide R in mobile phase A and
dilute to 10.0 mL with mobile phase A. Dilute 1.0 mL of
this solution to-l00.0 mL with mobile phase A.
Cefadroxil Monohydrate Reference solution (e) Dilute 1.0 mL of reference solution (c)
(Ph. Eur. monograph 0813) to 25.0 mL with mobile phase A.
Column:
- size: 1= 0.10 m, 0 = 4.6 mm,
- stationary phase: spherical octade<y/silyl siJiaz gelfor
MoMe phase:
- moMe phaseA: phosphate buffer soiution pH 5.0 R,
- mobile phase B: methanol R2,
381.4 66592-87-8
Action and use (min) (per cent 1//11) (per cent 1//11)
Cephalosporin antibacterial. 0-1 98 2
Preparations 1-20 98--->70 2 ---> 30
Cefadroxil Capsules
Cefadroxil Oral Suspension Flow rare 1.5 mUmin.
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2022 Cefadroxil Monohydrate 1-459
Detection Spectrophotometer at 220 urn. IMPURITIES

Injection 20 ~L of the test solution and reference
H NH2
solutions (c), (d) and (e).
Relative retention With reference £0 cefadroxil (retention ~CO'H
time = about 6 min): dimethylfonnamide = about 0.4;
dimethylaceramide = about 0.75.
HO~
System suitability: A. (2R)-2-amino-2-(4-hydroxyphenyl)acetic acid,
- resolution: minimum 5.0 between the peaks due £0
impurities A and B in the chromatogram obtained with
reference solution (c),
- signal-eo-noise ratio: minimum 10 for the 2nd peak in the
chromatogram obtained with reference solution (e).
Limits:
- impurity A: not more than the area of the I S ( peak in the
chromatogram obtainedwith reference solution (c) B. (6R,7R)-7-amino-3-methyl-8-oxo-5-thia-l-
(1.0 per cent), azabicyclo[4.2.0]oct-2-ene-2-earboxylic acid (7-ADCA),
- any other impu,ity: for each impurity, not more than the
area of the 2 nd peak in the chromatogram obtained with
reference solution (c) (1.0 per cent),
- total: not more than 3 times the area of the 2nd peak In
(3.0 per cent),
- disregard limit: 0.05 times the area of the 2nd peak in the
C. (2R,5RS)-2-[(R)-[[(2R)-2-amino-2-(4-
(0.05 per cent); disregard the peaks due to
hydroxyphenyl)acetyl]amino]carbuxymethyl]-5-methyl-5,6-
dimethylfcrmamide and dimethylacetamide.
dihydro-2H-I)3-thiazine-4-carboxylic acid,
N,N-Dlmethylanillne (2.4.26, Method B)
Maximum 20 ppm.
Water (2.5.12)
ASSAY
Liquid chromatography (2.2.29). D. (6R,7R)-7-[[(2S)-2-amino-2-(4-hydroxyphenyl)acetyl]
amino]-3-methyl-8-oxo-5-thia-I-azabicyclo[4.2.0] oct-2-
ene-2-earboxylic acid (r-cefadroxil),
the mobile phase.
Reference solution (a) Dissolve 50.0 mg of cefadroxil CRS in
phase.
Reference solution (b) Dissolve 5 mg of cefodroxll CRS and
50 mg of amoxicil/in trihydra" CRS in the mobile phase and
dilute to 100 mL with the mobile phase. E. (6RS)-3-(aminomethylene)-6-(4-hydroxyphenyJ)
Column: piperazine-Zfi-dione,
- size: 1= 0.25 m, 0 = 4.6 rom,
- stationary phase: octade<ylsi/yl silica gelfor chromatography R
(5 pm).
Mobile phase acetonitrile R, a 2.72 gIL solution of potassium
dihydrogen phosphate R (4:96 VIV).
Flow rate 1 mllmin.
Injection 20 ~L.
F. (6R,7R)-7-[[(2R)-2-[[(2RS)-2-amino-2-(4-
System suitability Reference solution (b): hydroxyphenyl) acetyl]amino]-2-(4-hydroxyphenyl)acetyl]
- resolution: minimum 5.0 between the peaks due to amino]-3-methyl-8-oxo-5-thia-l-azabicyclo[4.2.0] oct-2-
cefadroxil and to amoxicillin. ene-2-earboxylic acid,
Calculate the percentage content of cefadroxil.
HO
STORAGE
Protected from light. O~CH'
S
G. 3-hydroxy-4-methylthiophen-2(5H)-one,
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1-460 Cefalexin Monohydrate 2022

mobile phase A.
Reference solution (aJ Dissolve 10.0 mg of D-pheuylglycine R
in mobile phase A and dilute to 10.0 mL with mobile
phase A.
H. (6R,7R)-7 -I(2,2-<1imethylpropanoyl)amino]-3-methyl-8-
oxo-5-thia-I-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid Reference solution (b) Dissolve 10.0 mg of
(7-ADCA pivalamide). r-aminodesacaoxycepholosporanic acidCRS in phosphate buffer
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
solution pH 7.0 R5 and dilute to 10.0 mL with mobile
phase A.
and 1.0 mL of reference solution (b) to 100.0 mL with
mobile phase A.
Cefalexin Monohydrate Reference solution (d) Dissolve 10 mg of dimethylfont/amide R
(Ph. Eur. monograph 0708) and 10 mg of dimethylacetamide R in mobile phase A and
dilute to 10.0 mL with mobile phase A. Dilute 1.0 mL of
ZCH3
H. NH2~,~~:J
o
this solution to 100.0 mL with mobile phase A.
Reference solution (e) Dilute 1.0 mL of reference solution (c)
err
""-
I
0
't-t-s
H H
365.4
H20
23325-78-2
Reference solution (f) Dissolve 10 mg of cefotaxime
sodium CRS in mobile phase A and dilute to 10.0 mL with
mobile phase A. To 1.0 mL of this solution add 1.0 mL of
the test solution and dilute to 100 mL with mobile phase A.
Column:
Action and use ~ size: / = 0.10 m, 0 = 4.6 mm;
Cephalosporin antibacterial. - stationary phase: spherical octade<ylsilyl silica gelfor

chromatcgraphy R (5 pm}.
Preparations
Cefalexin Capsules Mobile phase:
- mobile phase A: phosphate buffer solution pH 5.0 R;
Cefalexin Oral Suspension
- mobile phase B: methanol R2;
Cefalexin Tablets
PhE" _ Tim. MobUe phase A Mobile phase B
(mln) (per cent VIJI) (per cent VIJI)
DEFINITION 0-1 98 2
(6R, 7KJ-7-[[(2KJ-2-Amino-2-phenylacetyl]amino]-3-methyl- 1-20 98 -> 70 2 -> 30
8-oxo-5-thia-l-azabicyclo[4.2.0]0ct-2-ene-2-carboxylic acid
monohydrate. Flow rate 1.5 mllmin.
Semi-synthetic product derived from a fermentation product. Detection Spectrophotometer at 220 om.
Content lnjeaion 20 ilL of the test solution and reference
95.0 per cent to 102.0 per cent (anhydrous substance). solutions (c), (d), (e) and (I).
CHARACTERS System suirabiliry:
Appearance ~ resolution: minimum 2.0 between the peaks due to
White or almost white, crystalline powder. impurities A and B in the chromatogram obtained with
Solubility reference solution (e) and minimum 1.5 between the
Sparingly soluble in water, practically insoluble in ethanol peaks due to cefalexin and cefotaxime in the
(96 per cent). chromatogram obtained with reference solution (t).
Limits:
IDENTIFICATION ~ jmpun"ty B: not more than the area of the 2nd peak in the
chromatogram obtained with reference solution (e)
Comparison cefalexm monohydrate CRS. (1.0 per cent);
TESTS ~ attY other impurity: not more than the area of the 1sr peak
pH (2.2.3) in the chromatogram obtained with reference solution (c)
4.0 to 5.5. (1.0 per cent);
~ total: not more than 3 times the area of the I" peak in the
Dissolve 50 mg in carbon dioxide-free water R and dilute {Q
(3.0 per cent);
Specific optical rotation (2.2.7) ~ disregard limit: the area of the 2nd peak in the
+ 149 to + 158 (anhydrous substance). chromatogram obtained with reference solution (e)
Dissolve 0.125 g inphrhalate buffer solution pH 4.4 Rand (0.05 per cent); disregard any peaksdue to
dilute to 25.0 mL with the same solvent. dimethylfonnamide or dimethylacetamide.
Related substances N,N-Dimethylaniline (2.4.26, Method B)
Liquid chromatography (2.2.29) .. Maximum 20 ppm.
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2022 Cefalo tin Sodium 1-461
Water (2.5.12)
Sulfated ash (2A.1f)
D.3-hyclroxy-4-memylthiophen-2(5H)-one,
ASSAY
examined in water R and dilute (0 100.0 mL with the same
solvent.
Reference solution (a) Dissolve 50.0 mg of cefalexin
monohydrate GRS in waterR and dilute to 100.0 mL with the E. (6R,7R)-7 -[(2,2-dimethylpropanoyl)amino]-3-methyl-8-
same solvent oxo-S-rhia-l-azabicyclo]4.2.0]oct-2-ene-2-carboxylic acid
Reference soituion (b) Dissolve 10 mg of cefradine GRS in (7-ADCA pivalamide),
20 mL of reference solution (a) and dilute to 100 mL with
water R.
Column: "', a ":i.
N",~_)---~, I
~
CO,"CH
3
and eprner al
cYr
C~
- size: / = 0.25 m, 0 = 4.6 mm; ,p I -j----} S
- suuionary phase: ocUJd«y/si/y1 silica gel for chromatography R H H
(5 urn). ~ a
iHobile phase methanol R, acetonitrile R, J3.6 gIL solution of
F. (2RS,6R, 7R)-7 -[[(2R)-2-amino-2-phenylacetyl]amino]-3-
potassium dihydrogen phosphate R, water R methyl-8-oxo-5-thia-l-azabicyclo[4.2.O]oct-3-ene-2-
(2:5:10:83 VIVIVIV).
carboxylic acid (delta-2-cefalexin).
Flaw rate 1.5 mUmin. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E"
Injection 20 J.lL.
System suitabl7ity Reference solution (b):
Cefalotin Sodium ***
*** ***
cefalexin and cefradine.
Calculate the percentage content of cefalexin monohydrate. (Ph. Bur. monograph 0987) ***
STORAGE
IMPURITIES
C";l!,,N,NaO.S, 418.4 51H1-9
Action and use

A. (2R)-2-amino-2-phenylacetic acid (D-phenylglycine), Cephalosporin antibacterial.
P1>E" _
a ~CO,H
, CH3 DEFINITION
",N-Hs Sodium (6R, 7R)-3-[(acetyloxy)methyl]-8-oxo-7-[(thiophen-2-

ylacetyI)amino] -5-thia-I-azabicyclo[4.2.0] oct-z-ene-z-
"" carboxylate.
B. (6R, 7R)-7 -amino-3-methyl-8-oxo-5-thia-I-azabicyclo
[4.2.0]oct-2-ene-2-<:arboxylic acid Content
(7-aminodesacetoxycephalosporanic acid, 7-ADCA),
CHARACTERS
Appearance
Solubility
Freely soluble in water, slightly soluble in anhydrous ethanol.
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.2f).
C. (6R, 7 R)-7-[[(2R)-2-[[(2R)-2-amino-2-phenylacetyl] Comparison cefawtin sodium CRS.
amino]-2-phenylacetyl]amino]-3-methyl-8-oxo-5-thia-l- B. It gives reaction (a) of sodium (2.3.1).
azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid,
TESTS
Solution S
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1-462 Cefalotin Sodium 2022
Appearance of solution Limits:

Solution S is clear (2.2.1) and its absorbance (2.2.25) at - ;mpun'ty B: not more than the area of the principal peak in
450 om is not greater than 0.20. the chromatogram obtained with reference solution (b)
pH (2.2.3) (1.0 per cent);
4.5 to 7.0 for solution S. - impun"lY D: not more than 0.5 times the area of the
- any other impun"ty: for each impurity, not more than
Dissolve 1.25 g in water R and dilute to 25.0 mL with the 0.25 times the area of the principal peak in tbe
same solvent. chromatogram obtained with reference solution (b)
Related substances (0.25 per cent);
Liquid chromatography (2.2.29). Prepare ,he solutions - total: not more than 3 times the area of the principal peak
immediately before use. in the chromatogram obtained with reference solution (b)
res' solution (a) Dissolve 75.0 mg of the substance to be (3.0 per cent);
examined in water R and dilute to 25.0 mL with the same - disregard limit: 0.05 times the area of the principal peak in
solvent. the chromatogram obtained with reference solution (b)
(0.05 per cent).
Test solution (b) Dilute 5.0 mL of test solution (a) to
50.0 mL with water R. N,N-Dimethylaniline (2.4.26, Method B)
Reference solution (a) Dissolve 75.0 mg of cefalotin Maximum 20 ppm.
sodium CRS in waterR and dilute to 25.0 mL with the same 2-Ethylhexanoic acid (2.4.28)
solvent. Dilute 5.0 mL of the solution to 50.0 mL with Maximum 0.5 per cent.
waterR. Water (2.5.12)
Reference solution (b) Dilute 1.0 mL oftest solution (a) to Maximwn 1.5 per cent, determined on 0.500 g.
100.0 mL with wa'er R. Bacterial endotoxins (2.6.14)
Reference sdution (c) Mix 1 mL of test solution (a), I mL of Less than 0.13 IU/mg, if intended for use in the manufacture
hydrochloric acidRl and 8 mL of warer R. Heat at 60 °C for of parenteral preparations without a further appropriate
12 min and cool to room temperature in iced water. Inject procedure for the removal of bacterial endotoxins.
immediately.
ASSAY
Reference solution (d) Dissolve 5 mg ofcefalolinfor Liquid chromatography (2.2.29) as described in the test for
impurity B identification CRS.in waterR and dilute to 5 mL related substances with the following modifications.
Mobile phase Mix 14 volumes of autonitn1e Rand
Column: 86 volumes ofa 6.967 gIL solution of dipowsium hydrogen
- size: 1= 0.25 ID, 0 = 4.6 nun; phosphate R previously adjusted to pH 6.0 with phosphoric
- stationary phase: end-capped oetade<ylsi/yl sHi",gelfor acidR.
chromatography R (5 ~m);
- temperature: 40°C. Detection Spectrophotometer at 260 om.
Mob.e phase: Injection 5 J1L of test solution (b) and reference solution (a).
- mobile phase A: mix 3 volumes of acetonitrile Rl and Run lime 1.5 times the retention time of cefalotin (retention
97 volumes of a 1.742 gIL solution of dipotassium hydrogen =
time about 10 min).
phosphate R previously adjusted to pH 2.5 with phosphoric Calculate the percentage content of CIJ-IlsN2Na06S2 using
acidR; the chromatogram obtained with reference solution (a) and
- mobile phase B: mix 40 volumes of acetonitrile Rl and taking into account the assigned content of cefalotin
60 volumes of a 1.742 gIL solution of dipotassium hydrogen sodium CRS.
phosphate R previously adjusted to pH 2.5 with phosphoric STORAGE
acidRj
Protected from light. If me substance is sterile, store in a
Time Moblle phase A MobUe phose B
(mlo) (per cent JIm (per cent V/I1 IMPURITIES.
o ~ 30 100 ..... 0 0--+100 Specified impurities B, D.
30 - 35 0 100 Other detectable impuri,ies (thefollowing substonces would, if
present al a sufficienl level, be deteaed by oneor other of the rests
Flow rate 1.0 mUmiIl. in lhe monograph. They arelimited by the general aaepumce
Detection Spectrophotometer at 220 nm. criterion for other/unspecified impurities and/or fry thegeneral
monograph Substances for phannaceuticol use (2034). I, is
Injection 20 ....L of test solution (a) and reference
Relative retention With reference to cefalotin (retention in substances for pharmaceutical use) A, C.
time = about 26 min): impurity C about 0.2; =
=
impurity B about 0.7; impurity D = about 0.88;
impurity D and cefalotin.
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2022 Cefamandole Nafate 1-463
carboxylate and sodium (6R,7R)-7-[[(2R)-2-hydroxy-2-

phenylacetyI]amino]- 3- [[( I-methyl-I H-te trazol-5-yI)sulfanyl)
methyl]-8-oxo-5-thia -I-aza bicyclo [4.2.0] oct-2-ene-2-
carboxylate (cefamandole sodium), with sodium carbonate.
A. (6R,7R)-3-methyl-8-oxo-7-[(thiophen-2-ylacetyl)amino]-5- Content
thia -1-azabicyclo [4.2.0Joct-z-ene-z-carboxylic acid - cefamandok nafate (C,.H 17NoNaO.S,): 93.0 per cent to
(deaceroxycefalotin), 102.0 per cent (anhydrous and sodium carbonate-free
substance), for the sum of the content of cefamandole
nafate and cefamandoJe sodium expressed as cefamandole
nafate;
- cefamandole sodium (CISH17N6NaOsSv: maximum
10.0 per cent (anhydrous and sodium carbonate-free
substance);
- sodium carbonate (Na2C03): 4.8 per cent to 6.4 per cent.
B. (6R,7R)-3-(hydroxymethyl)-8-oxo-7-[(thiophen-2-ylacetyl)
amino]-5-thia-l-azabicyclc[4.2.0] oct-2-ene-2-carboxylic CHARACTERS
acid (deacerylcefatorin), Appearance
SolubUlty
Freelysoluble in water, sparingly soluble in methanol.
IDENTIFICATION
C. (6R,7R)-3-[ (acetyloxy)methyl)-7-amino-8-oxo-5-thia-l- Comparison ufamandole nafiue CRS.
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (7-ACA), B. It gives reaction (a) of sodium (2.3.1).
TESTS
Solution S
Dissolve 2.5 g in carbon dioxide-fret waterR and dilute to
Solution S is clear (2.2.1) and its absorbance (2.2.25) at
475 nm is not greater than 0.03.
D. (5aR,6R)-6-[(thiophen-2-ylacetyl)amino]-5a,6-dihydro-
pH
3H, 7H-azeto[2, l-b]furo[3,4-d] [I ,3]thiazine-l,7(4H)-dione
6.0 to 8.0 for solution S, measured after 30 min.
(cefalotin lactone).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" Specific optical rotation (2.2.7)
-35.0 to -45.0 (anhydrous and sodium carbonate-free
substance).
Dissolve 1.00 g in acetate buffersolution pH 4.7 Rl and dilute
Cefamandole Nafate
Related substances
(Ph. Eur. monograph 1402) Liquid chromatography (2.2.29). Prepare the solutions
Solvent mixture Mix 18 volumes of acetonitrile Rand
75 volumes of a 10 per cent VIV solutionof triethylamine R
previously adjusted to pH 2.5 with phosphonc acid R.
examined In the solvent mixture and dilute to 10.0 mL with
Compound R Molecular Fomlula Mr
Reference solution (a) Dilute 1 mL of the test solution to
Cefamandole nefate CHO CI!lillN6Na06~ 512.5
10 mL with the solvent mixture, then heat at 60°C for
Cefamandolesodium H C18H'lNaNaOsSz 484.5 30 min.
Cefamandole nafate 42540-40-9 100.0 mL with the solvent mixture.
Cefamandole sodium 30034-03-8 Column:
-- size: I;;;:; 0.25 m, 0 = 4.6 mm;
Action and use - stationary phase: ocladecylsilyl silica gd for chromatography R
Cephalosporin antibacterial. (5 urn).
PhE" _ Mobile phase:
- triethylamine phosphate solution: dissolve 2.0 g of sodium
DEFINITION pentanesulfonale R in 350 mL of waterR, add 40 mL of
Mixture of sodium (6R,7R)-7-[[(2R)-2-(formyloxy)-2- triethylamine R, adjust to pH 2.5 with phosphoric acidR
phenylacetyl] amino]-3- [[(l-methyl-IH-tetrazol-5-yl)sulfanyl] and dilute to 700 mL with waur R;
methyl]-8-oxo-5-thia-I-azabicyclo[4.2.0] oct-2-ene-2-
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1-464 Cefamandole Nafate 2022
- mobile phase A: mix 1 volume of the triethylamine - repeatability: maximum relative standard deviation of
phosphate solution and 2 volumes of water Rj 0.8 per cent after a series of single injections of not less
- mobile phase B: mix equal volumes of the triethylamine than 3 freshly prepared reference solutions (a).
phosphate solution, methanol R and acetonitrile Rj Calculate the percentage content of cefamandole nafate
(C 19H 17No;NaO.S,j from the sum of the contents of
Time Mobile phase A Mobile phase B cefamandole nafate and cefamandole sodiumexpressed as
(min) (per cent VII') (pel'" cent VIJI)
cefamandole nafate, using the declared content of cefamandole
0-1 100 0 nafate CRS.
1·35 100 -+ 0 o -> 100
1 mg of cefamandole sodium is equivalent to 1.0578 mg of
cefamandole nafate.
Sodium carbonate
Detection Spectrophotometer at 254 nm. Dissolve 0.500 g in 50 mL of water R. Titrate with 0.1 M
Injection 20 JlL loop injector. hydrochlom acid, determining the end-point potentiometrically
Relative retention with reference to cefamandole nafate (2.2.20).
(retention time = about 24 min): cefamandole = about 0.8. 1 mL of 0.1 M hydrochloric acid is equivalent to 5.3 mg uf
System suitability Reference solution (a): Na,CO,.
- resolution: minimum 5.0 between the peaks due to STORAGE
cefamandole and cefamandole nafate. In an airtight container, protected from light. If the substance
Limits: is sterile, store in a sterile) airtight, tamper-evident container.
- artY impurity: for each impurity, not more than the area of
LABELLING
reference solution (b) (1.0 per cent); The labelstates that the substance contains sodium
- total: not more than 5 times the area of the principal peak carbonate.
in the chromatogram obtainedwithreference solution (b) IMPURITIES
(5.0 per cent);
(0.1 per cent).
2-Ethylhexanolc acid (2.4.28)
Maximwn 0.3 per cent mlm.
Water (2.5.12)
Maximum 2.0 per cent, determined on 0.500 g. A. (6R,7R)-7-[[(2R)-2-(fonnyloxy)-2-phenylacetyl]amino]-3-
methyl-8-oxo-5-thia-I-azabicyclo[4.2.01Oct-2-ene-2-
carboxylic acid (formylmandeloyl-7-amino-desacetoxy-
Less than0.15 IU/mg J if intended for use in the manufacture
cephalosporanic acid),
ASSAY
Cefamandole nafate
examined in the mobilephase and dilute to 100.0 mL with C. (6R, 7R)-7 -[[(2R)-2-(acetyloxy)-2-phenylacetyl]amino]-3-
the mobile phase. [[(l-methyl-IH-tetrazol-5 -yl)sulfanyl] methylj-8-oxo-5-
Reference solution (a) Dissolve 50.0 mg of cefamandole thia-I-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (0-
nafate CRS in the mobile phase and dilute to 100.0 mL with acetylcefamandole),
the mobilephase.
Reference solution (b) Dilute 1 mL of the test solution to
10 mL with the mobile phase, then heat at 60 "C for 30 min.
Column:
- size: 1= 0.25 rn, 0 = 4.6 mm;
- stationary phase: ocwde<ylsilyl silica gelfor chroma"'graphy R
D.I-methyl-IH-tetrazole-5-thiol,
(5 urn).
Mobile phase Mix 25 volumes of _",nitrile R and
75 volumes of a 10 per cent VIV solution of triethylamine R
previously adjusted to pH 2.5 with phosphoric acid R.
Deration Spectrophotometer at 254 run.
Injection 20 ~L loop injector.
System suitability: E. (6R, 7 R)-7 -[[(2R)-2-(fonnyloxy)-2-phenylacetyl]aminoj-3-
- resolution: minimum 7.0 between the 2 principal peaks in [(acetyloxy)methyl] -8-oxo-5-thia-I-azabicyclo[4.2.0] oct-2-
the chromatogram obtained with reference solution (b); ene-2-carboxylic acid (fonnylmandeloyl-7-ACA).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E"
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2022 Cefapirin Sodium 1-465
Cefapirin Sodium *** Column:

*** *** - size: / = 0.30 m, 0 = 4 mm,
(Ph. Bur. monograph 1650) *** - stationary phase: octadeqlsily/ silica gd for chromatography R
(10 pm).
MoMe phase Mix 80 mL of dimerhylfonnamide R, 4.0 mL of
glacial acetic add Rand 20 mL of a 4.5 per cenr mlm solution
of potassium hydroxide R. Dilute to 2 L with warer R.
445.5 24355-60-3 Injection 20 J.lL of the test solution and reference
Action and use Run time Twice the retention time of cefapirin.
Relativeretention With reference to cefapirin (retention
PhE" - -_ _ time = about 13 min): impurity B = about 0.3;
impurity C = about 0.5; impurity A = about 0.75.
DEFINITION
System SU'jtabiJi~ Reference solution (d):
Sodium (6R,7R) -3-[(acetyloxy)methyl)-8-oxo-7-[ [[(pyridin-4-
yl)sulfanyl)acetyl)amino)-5-thia-I-azabicyclo[4.2.0) oct-2-ene-
cefapirin and impurity A.
2-carboxylate.
Limits:
--.,.- any impuniy: for each impurity, not more than the area of
Content the principal peak in the chromatogram obtained with
96.0 per cent to 102.0 per cent (anhydrous substance). reference solution (b) (1.0 per cent), and not more than
CHARACTERS 1 such peakhas an area greater than 0.3 times the area of
Appearance the principal peak in the chromatogram obtained with
White or pale yellow powder. reference solution (b) (0.3 per cent),
Solubility
Soluble in water, practically insoluble in methylene chloride. (2.0 per cent),
IDENTIFICATION - disregard limit: area of the principal peak in the
A. Infrared absorption spectrophotometry (2.2.24). chromatogram obtained with reference solution (c)
Comparison cefapin"n sodium CRS. (0.05 per cent).
B. It gives reaction (a) of sodium (2.3.1). N,N-Dlmethylaniline (2.4.26, Merhod B)
Maximum 20 ppm.
TESTS
Appearance of solution 2-Ethylliexanolc acid (2.4.28)
Dissolve 2.0 g in water R and dilute to 10.0 mL with the Maximum 0.5 per cent.
same solvent The solutionis clear (2.2.1). Dilute 5.0 mL to Water (2.5.12)
10.0 mL with water R. The absorbance (2.2.25) of this Maximum 2.0 per cent, determined on 0.300 g.
solution at 450 nm is not greater than 0.25. Bacterial endotoxlns (2.6.1 <f)
pH (2.2.3) Less than 0.17 IV/mg, if intended for use in the manufacture
6.5 to 8.5. of parenteral preparations without a further appropriate
Dissolve 0.100 g in carbon dioxide-free water R and dilute to procedure for the removal of bacterial endotoxins.
10.0 mL with the same solvent. ASSAY
Specific optical rotation (2.2.7) Liquid chromatography (2.2.29) as described in the tesr for
+ 150 to + 165 (anhydrous substance). related substances with the following modification.
Dissolve 0.500 g in water R and dilute to 25.0 mL with the Injection Test solution and reference solution (a).
same solvent. Calculate the percentage content of C17Hl~3Na06S2'
Related substances STORAGE
Liquid chromatography (2.2.29). Prepare the solutions Protected from light. If the substance is sterile, store in a
immediacely before use. sterile, tamper-evident container.
IMPURITIES
the mobile phase.
Reference solution (a) Dissolve 42 mg of cefapirin
sodium CRS in the mobile phase and dilute to 200.0 mL with
the mobile phase.
Reference solution (c) Dilute J.O mL of reference solution (b)
A. (5aR,6R)-6-[[[(pyridin-4-yl)sulfanyl)acetyl)amino)-5a,6-
Reference solution (d) Mix I mL of the test solution, 8 mL
dihydro- 3H, 7H-azeto[2, l-b)furo[3,4-d] [1,3)thiazine-l,7
of the mobile phase and I mL of hydrochloric acid R1. Heat
(4H)-dione (deacetylcefapirin lactone),
at 60 °C for 10 min.
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1-466 Cefatrizine Propylene Glycol 2022

TESTS
B. (6R,7R)-3-(hydroxymethyl)-8-oxo-7-([[(Pyridin-4-yl)
sulfanyl]acetyl]amino)-5-thia-l-azabicyclo [4.2.0)oct-2-ene-
2-carboxylic acid (deacerylcefapirin), Dissolve 0.400 g in 1 M hydrochloric add and dilute to
Propylene glycol
Solvent mixture ace"'ne R, water R (20:80 VII').
Internal standard solution Dissolve 1.0 g of
dimethylaceramide R in the solvent mixture and dilute to
C. (6R,7R)-3-methyl-8-oxo-7-([[(Pyridin-4-yl)sulfanyl)acetyl] Test solution Introduce 0.40 g of the substance to be
amino]-5-thia-l-azabicyclo[4.2.01oct-2-ene-2-carboxylic examined into a ground-glass-stoppered test-tube.
acid (deacetoxycefapirin). Add 3.0 mL of the internal standard solution) 1.0 rnL of the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ !'hE" solvent mixture and 2.0 mL of hydroehlon'c acid R. Seal the
test-tube and shake.
Reference solution (a) Dissolve 2.0 g of propylene glYcol R in
Cefatrizine Propylene Glycol ****
***
mixture.
*
(ph. Eur. monograph 1403) **** Reference solution (b) Introduce into a ground-glass-
stoppered test-tube 1.0 mL of reference solution (a) and
1.0 mL of the internal standard solution.
Column:
- size: 1= 2 m, 0 = 2 rom;
- stationary phase: ethylvinylbenzene-divinylbenzene
cupo!Ymer R (150-180 urn).
Corner gas nitrogen for chromatography R.
Flow rau 30 mUmin.
Temperature:
- column: 200 "C;
C 18H,sN,O,S,,(C,HaO,). 462.5 - inje<n'on port and detector: 250 "C.
(base)
Action and use Inieaion 1 J.lL of the test solution and reference solution (b).
Cephalosporin antibacterial. Limit:
!'hE" _
- propylene glYcol: 13.0 per cent to 18.0 pee cent.
Related substances
DEFINITION Liquid chromatography (2.2.29).
Mixtute of (6R,7R)-7-[[(2R)-2-amino-2-(4- Test solution Dissolve 60.0 mg of the substance to be
hydroxyphenyl)acetyl) amino)-8-oxo-3-[[(IH-I,2,3-trlazol-4- examinedin the mobile phase and dilute to 100.0 mL with
yl)sulfanyl]methyl]-5-thia-l-azabicyclo[4.2.0) oct -z-ene-z- the mobile phase.
carboxylic acid and propane-l,2-diol in molecular
Reference solution (a) Dissolve 60.0 mg of eefatrizine
proportions of about 1:1.
propylene glYcol CRS in the mobile phase and dilute to
Content 100.0 mL with the mobile phase.
95.0 per cent to 102.0 per cent of C18H18N605~
Reference solution (b) Dissolve 30.0 mg of eefatrizine
(anhydrous substance).
impurity A CRS in buffer solution pH 7.0 R and dilute to
CHARACTERS 100.0 mL with the same buffer solution.
Appearance Reference solution (c) Dilute 0.6 mL of reference solution (a)
White or almost white powder. to 100.0 mL with the mobile phase.
Solubility Reference solution (d) Dilute 1.0 mL of reference
Slightly soluble in water, practically insoluble in ethanol solution (b) to 100.0 mL with buffer solutian pH 7.0 R.
(96 per cent) and in methylene chloride. Reference solution (e) To 1.0 mL of reference solution (a)
IDENTIFICATION add 1.0 mL of reference solution (b) and dilute to 10.0 mL
A. Infrared absorption spectrophotometry (2.2.24). with the mobile phase.
Comparison eefatrizine propylene glYcol CRS. Column:
B. Examine the chromatograms obtained in the test for - size: 1 = 0.25 m, 0 = 4 rom;
propylene glycol.
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2022 Cefazolin Sodium 1-467
*****
- stationary phase: octadecylsiiyl silica gelfor chromatography R
(5 urn).
Cefazolin Sodium
* *
Mobile phase Mix 5 volumes of acetonitrile Rand (Ph. Ellr. monograph 0988)
**** *
phosphate R in water R.
Flow rate 2 mllrnin.
solutions (e), (d) and (e).
Run time At least twice the retention time of cefatrizine.
System SUiUlbility Reference solution (e):
C,Jf"N,NaO,S, 476.5 27164-46-1
cefatrizine and impurity A. Action and use
Limits: Cephalosporin antibacterial.
- impurity A: not more than the area of the corresponding Preparation
Cefazolin Injection
solution (d) (0.5 per cent);
- any otherimpurity; for each impurity, not more than the PhE" _
area of the principal peak: in the chromatogram obtained
DEFINITION
with reference solution (c) (0.6 per cent);
Sodium (6R, 7R)-3-[[(5-methyl-I,3,4-thiadiazol-2-yl)sulfanyl]
- sum of impurities other than A: not more than 3.5 times the
methyl] -8-oxo-7- [( IH-tetrazol-I-ylacetyl)amino]-5-thia-l-
area of me principal peak in me chromatogram obtained
azabicyclo[4.2.0]oct-2-ene-2-carboxylate.
- disregard limit: 0.05 times the area of the principal peak in Semi-synthetic product derived from a fermentation product.
me chromatogram obtained with reference solution (c) Content
(0.03 per cent). 95.0 per cent to 102.0 per cent (anhydrous substance),
Water (2.5.1Z) CHARACTERS
Maximum 1.5 per cent, determined on 0.500 g. Appearance
Sulfated ash (2.4.14) White or almost white powder, very hygroscopic.
Maximum 0.1 per cent, 'determined on 1.0 g. Solubility
ASSAY Freely soluble in water, very slightly soluble in ethanol
Liquid chromatography (2.2.29) as described in the test for (96 per cent).
related substances with the following modifications. It shows polymorphism (5.9).
Injection Test solution and reference solution (a). IDENTIFICATION
System suitability Reference solution (a): A. Infrared absorption spectrophotometry (2.2.24).
- repeatabih'ty: maximum relative standard deviation of Preparation Dissolve 0.150 ginS mL of water R, add
1.0 per cent after 6 injections. 0,5 mL of dl1ute acetic acid R, swirl and allow to stand for
Calculate the percentage content of CisHlSN60jS2 from the 10 min in iced water. Filter the precipitate and rinse with
declared content of CisHlaN60jS2 in ofameinepropylene 1-2 mL of water R. Dissolve in a mixture of 1 volume of
ery",,1 CRS. water Rand 9 volumes of acetone R. Evaporate me solvent
IMPURITIES almost to dryness, then dry in an oven at 60 °C for 30 min.
Specified impurities A. Comparison ceJazolin CRS.
B. It gives reaction (a) of sodium (2.3.1).
a yeO'H TESTS
H,N tls
N ""
~,~
S
N-NH
Solution S
Solution S is clear (2.2.1) and its absorbance (2.2.25) at
A. (6R, 7R)-7 -amino-s-oxo-3-[[(lH-I,2,3-triazol-4-yl) 430 nm is not greater than 0,15.
sulfanyl] methyl]-5-thia-l-azabicyclo[4.2.0] oct-2-ene-2-
carboxylic acid (7-ACA triazole). pH (2.2.3)
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
-24 to -15 (anhydrous substance).
Dissolve 1.25 g in waterR and dilute to 25.0 mL with the
same solvent.
Absorbance (2.2.25)
with sodium hydrogen carbonate solution R. Examined between
220 om and 350 om, the solution shows an absorption
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1-468 Cefazolin Sodium 2022
1
r I ~I .I I I I I I I I I I I I J
0 1.3 2.5 3.8 5.0 6.3 7.5 8.8 10.0 11.3 12.5 13.8 15.0 16.3 17.5 18.8 min
1. impurity J 2. impurity E 3. unknown structure 4. cefazolin 5. impurity L
Figure 0988.-1. - Onromatogrcm for the test for related subslances of ufazolin sodium: reference solution (b) (in situ degradation)
maximum at 272 nm. The specific absorbance at the Flow rate I. 2 mUmin.
maximum is 260 to 300 (anhydrous substance). Detection Spectrophotometer at 254 om.
Related substances I nj",... 5 pL.
Liquid chromatography (1.1.19). System suitability Reference solution (b):
Test solution Dissolve 50.0 mg of the substance to be - resolution: minimum 2.0 between the peaks due to
examined in mobile phase A and dilute to 20.0 mL with cefazclin and impurity L (see Figure 0988.-1).
mobile phase A. Limits:
Reference ,00ution (a) Dilute 1.0 mL of the test solution to - any impun"ty: for each impurity) not more than the area of
100.0 mL with mobile phase A. the principal peak in the chromatogram obtained with
Reference solution (b) Dissolve 20 mg of the substance to be reference solution (a) (l.0 per cent);
examined in 10 mL uf a 2 gIL solution of sodium hydroxide R. - uxal: not more than 3.5 times the area of the principal
Allow £0 stand for 15-30 min. Dilute 1.0 mL of the solution peak in the chromatogram obtained with reference
to 20 mL with mobile phase A. solution (a) (3.5 per cent);
Column: - disregard limit: 0.05 times the area of the principal peak in
- size: 1 = 0.125 ffi l 0 = 4.0 mm, the chromatogram obtained with reference solution (a)
- 'tationary phase: end-eapped oaad",ylsilyl sitica gelfor (0.05 per cent).
chromalOgraphy R (3 pm); N,N-Dlrnethylaniline (1.4.26, Method B)
- temperature: 45 "C. Maximum 20 ppm.
Mob!7e phase: Water (2.5.IZ)
- mobile phaseA: solution containing 14.54 gIL of disodium Maximum 6.0 per cent) determined on 0.300 g.
hydrogen phosphare dlJfl.ecahydrate Rand 3.53 gIL of Bacterial endotoxins (1.6.14)
potassium dihydrogen phosphare R; Less than 0.15 ill/mg, if intended for use in the manufacture
- mobile phaseB: acelOnicrile R; of parenteral preparations without a further appropriate
procedure for the removal of bacterial endcroxins.
(min) (per cent VIV) (per cent VII-') ASSAY
0-2 98 2 Liquid chromatography (1.1.19).
2-' 98 ..... 85 2 ..... 15 Test solution Dissolve 50.0 mg of the substance to be
4 - to 85 ---> 60 15 ---> 40 examined in the mobile phase and dilute to 50.0 mL with
10 - U.5 60 ---> 35 40 ---> 65 the mobile phase.
11.5-12 35 65
Reference solution (a) Dissolve 50.0 mg of cefazolin CRS in
12 - 15 35 ---> 98 65 ---> 2
15 ~ 21 98 2 phase.
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2022 Cefazolin Sodium 1-469
Reference solution (b) Dissolve 5.0 mg of cefuroxime

sodium CRS in 10.0 mL of reference solution (a) and dilute
Column:
- size: I = 0.25 ID, (2) = 4.6 mm;
- stationary phase: end-eapped oaade<y/silyl silica gelfor
chromatography R (5 urn), c. (6R, 7R)-3-methyl-8-oxo-7-[(IH-tetrazol-I-ylacetyl)
amino]-5-rhia-f-azabicydo[4.2.0] oct-z-ene-z-cerboxyllc
Mobile phase Mix 10 volumes of acetonitrile Rand
90 volumes of a solution containing 2.77 gIL of disodium
acid,
hydrogen phosphate dodecahydrate Rand 1.86 gIL of cilric acid
monohydrate R.
Injection 20 ~L.
System suitability:
- resolution: minimwn 2.0 between the peaksdue [Q D. (6R, 7R)-3-[(acetyloxy)methyl]-8-oxo-7-[(IH-retrazol-l-
cefazolinand cefuroxime in the chromatogram obtained ylacetyI)am ino]-5-thia-l-azabicyclo[4.2.0]oCl-2-ene-2-
with reference solution (b); carboxylic acid,
- symmeny factor. 0.8 to 3.0 for the peak due to cefazolin in
Calculate the percentage content of C14H13NsNa04S3 taking
into account the assigned content of cejazolin CRS and a
conversion factor of 1.048.
STORAGE
In an airtight container, protected from light. If the substance E. 5-methyl-I,3,4-thiadiazol-2-thiol (MMTD),
is sterile, the container is also sterile and tamper-evident"
IMPURITIES
Otherdetectable impurities {thefollowing substances would, if
present at a sufficient level) bedetected by oneor other of the tests
criterion for other/unspecified impwities. II is therefore nol
necessary to i'dentify these impurities for demonstration of
compliance. See also5.10. Control of impurities in substances for
pharmaceutical use) A, B, C D, E, G, H, 1, J, s; L. G. (5aR,6R) -6- [(1H-tetrarol-I-ylacetyl)amino]-5a,6-dihydro-
j
3H,7H-azeto[2 ,1-b]furo[3,4-d] [I ,3Jthiazine-l,7( 4H)-<lione,
o yCo,H
H,N}~'
,··~s
'"N
AS
S ~N
H H \_'
H,Cr
N
H. (6R,7 R)-3-[ (acetyioxy)methylJ-7-amino-8-oxo-5-thia-l-

A. (6R, 7R)-7 -amino-3-[[(5-methyl-I,3,4-thiadiazol-2-yl) azabicyclo[4.2.0]oet-2-ene-2-carboxylic acid (7-ACA),
sulfanylJmethylJ-a-oxo-5-thia-l-azabicyclo[4.2.0] oct-2-
ene-2-carboxylic acid,
I. 2- [carboxy]( 1H-retrazol-l-ylacetyl)amino Jmethyl]-5-[[(5-

methyl-I,3,4-thiadiazol-2-yl)sulfanyl)methyl]-5,6-dihydro-
B. (6R, 7R)-7 -[(2,2-<1imethylplOpanoyl)amino]-3-[[(5-methyl- 2H-l,3-thiazille-4-earboxylic acid (cefazoloic acid),
1,3,4-thiadiazol-2-yl)sulfanylJmethylJ-8-oxo-5-thia-l-
azabicyclo[4.2.0]oet-2-ene-2-catboxylic acid,
1. 2- [carboxy[(I H-tetrazol-I-ylacetyI)amino[methyl]-5-
methylidene-5 J6-dihydro-2H-I,3-thiazine-4-earboxylic
acid,
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1-470 Cefepime Hydrochloride Monohydrate 2022
TESTS
than reference solution Y3 (2.2.2, Method If).
solvent.
Speclfic optical rotation (2.2.7)
K. (6R,7R}-3-[[(5-methyl-I,3,4-thiadiazol-2-yl}sulfanyl] + 40 to + 45 (anhydrous substance).
methyl]-B-oxo-7-[(IH-tetrazol-I-ylacetyl)amino]-5 -thia-I- Dissolve 0.250 g in warer R and dilute to 25.0 mL with the
azabicyclo[4.2.0]oct-2-ene-2-carboxamide same solvent.
(cefazolinamide),
Impurity G
immediately be/ore use.
TeS! solution Dissolve 0.100 g of the substance to be
examinedin 0.01 M nitric acid and dilute to 10.0 mL with
the same acid.
Reference solution (a) Dilute 0.250 g of N-methylpyn-olidine R
(impurity G) to 100.0 mL with warer R. Dilute 2.0 mL of
L. (6R,7S)-3-[[(5-methyl-I,3,4-thiadiazol-2-yl}sulfanyl] this solution to 100.0 mL with 0.01 M mine acid.
methyl]-B-oxo-7- [(1H-tetrazol-I-ylacetyl)amino]-5-thia-l- Reference solution (b) Dilute 0.250 g of f>YIWlidine R to
azabicyclo[4.2. Oloct-z-ene-z-cerboxyhc acid 100 mL with 0.01 M tlitn·, add. Dilute 2 mL of the solution
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" to 100 mL with 0.01 M nitric acid. Mix 5 mL of this solution
with 5 mL of reference solution (a).
Column:
- size: I = 0.05 m, 0 = 4.6 mm;
Cefepime Hydrochloride **** - stationary phase: strong cation-exchange resin R (5 JIm).
*** *
*** *
lWobiJe phase .M.ix 1 volume of acetonitrile Rand
Monohydrate 100 volumes of 0.01 M nitric acid; filter through a 0.2 pm
(Cefepime Dihydrochloride Manohydrate, Ph. Bur. filter.
manograph 2126) . Flow rate I mUmin.
Detection Conductivity detector.
Injection 100 pL.
Run time 1.1 times the retention time of cefepime.
Retention time Cefepime = about 50 min, elutingas a
broadened peak.
System suitabl7ity:
- symmetry factor: maximum 2.5 for the peak due to
571.5 123171-59-5 impurity G in the chromatogram obtained with reference
solution (a)j
Action and use
5.0 per cent after 6 injections of reference solution (a);
£'hE" _ - peak-to-valley ratio: minimum 3 between the peaks due to
pyrrolidine and impurity G in the chromatogram obtained
DEFINITION with reference solution (b).
(6R,7R}-7-[[(2Z)-(2-Aminothiazol-4-yl}(methoxyimino)
Calculate the percentage content of impurity G in the test
acetyl]amino]-3-[(J-methylpyrrolidlnio}methyl]-B-oxo-5-thia-
solution usingreference solution (a).
l-azabicyclo[4.2.0]oct-2-ene-2-carboxylate dihydrochloride
monohydrate. Semi-synthetic productderived from a Limit:
fermentation product. - impuriry G: maximum 0.5 per cent.
Content Related substances
97.0 per cent to 102.0 per cent (anhydrous substance). liquid chromatography (2.2.29). Prepare the solutions
immediately befo", use or keep refrigerated at 4-8 'C for not more
CHARACTERS than 12 h.
Appearance
examined in mobilephaseA and diluteto 50.0 mL with
Solubility mobile phaseA. Sonicatefor 30 s and stir for about 5 min.
Freely soluble in water and in methanol, practically insoluble
Referenu solution (a) Dissolve 70.0 mg of cefepime
in methylene chloride.
dihydrochlcn'de monohydrate CRS in mobile phase A and dilute
IDENTIFICATION to 50.0 mL with mobile phase A. Sonicate for 30 s and stir
A. Infrared absorption spectrophotometry (2.2.24). for about 5 min.
Comparison cefepime dihydrochlcride monohydrate CRS. Reference solution (b) Dilute 1.0 mL of the test solution to
B. It gives reaction (a) of chlorides (2.3.1). 10.0 mL with mobile phase A. Dilute 2.0 mL of this solution
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2022 Cefepirne Hydrochloride Monohydrate 1-471
Reference solution (c) Dissolve 7 mg of cefepime chromatogram obtained with reference solution (b)
dihydrochloride monohydrate for system suitability CRS (0.10 per cent);
(containing impurities AJ Band F) in mobile phase A and - total: not more than 5 times the area of the principal peak
dilute to 5 mL with mobile phase A. in the chromatogram obtained with reference solution (b)
Reference solution (d) Dissolve 2 mg of cefepime (1.0 per cent);
impurity E CRS in mobile phase A and dilute to 25.0 mL - disregard limit: 0.25 times the area of the principal peakin
with mobile phase A. Dilute 1.0 mL of the solution to the chromatogram obtainedwith reference solution (b)
10.0 mL with mobile phase A. (0.05 per cent).
Column: Water (2.5.12)
- size: 1= 0.25 m, 0 = 4.6 mm; 3.0 per cent to 4.5 per cent, determined on 0.400 g.
- stationary phase: end-capped o<radecylsi/yl silica gel/or Bacterial endotoxin. (2.6.14)
chromatography R (5 urn), Less than 0.04 ill/mg) if intended for use in the manufacture
Mobile phase: of parenteral preparations without a further appropriate
- mobile phase A: mix 10 volumes of acetonitrile Rand procedure for the removal of bacterial endoroxins.
90 volumes of a 0.68 gIL solution of potassium dihydrogen ASSAY
phasphate R previously adjusted to pH 5.0 with a 0.5 M
Liquid chromatography (2.2.29) as described in the test fer
potassium hydroxide solution prepared from potassium
hydroxide R;
- mobile phase B: mix equalvolumes of acetonitrile R and a Mobile phase Mobile phase A.
0.68 gIL solution of potassium dihydrogen phosphate R Injection Test solution and reference solution (a).
previously adjusted to pH 5.0 with a 0.5 M potassium R,m rime 1.4 times the retention time of cefepime.
hydroxide solutionprepared from potassium hydroxide Rj Calculate the percentage content of C19H26CbN605S2 from
the declared content of cefejnme dihydrIXh/aride
Time Mobile phase A Mobile phase B monohydrate CRS.
(min) (per cent VII') {per cent V/l?
0-10 100 0 STORAGE
10 - 30 100 -+ 50 0--->50 Protected from light. If the substance is sterile) store in a
30 - 35 50 50 sterile, airtight) tamper-evident container.
35·36 50 ..... 100 50 ..... 0 IMPURITIES
36 - 45 100 0 Specified impun"ties A, B, E, F, G.
Otherdetectable impumies (thefollowing substances would, if
Flow rate 1 mUmin. present at a sufficientleveJ, be detected by oneor other of the tests
Detection Spectrophotometer at 254 nm. in the monograph. They are limited by the general acceptance
Injection 10 ~L of the test solution and reference ctitetion for other/unspecified impun·ties and/or by thegeneral
solutions (b), (c) and (d). monograph Subsrancesfar pharmaceutical use (2034). 11 is
Identification of impuniies Use the chromatogram supplied therefore not nuessary to identify these impurities for
with cefepime dihydrlXhloride monohydrate fur system
demonstration of compliance. See also 5.10. Control of impuriues
in substances for pharmaceutical use) c) D.
suitability GRS and the chromatogram obtained with
reference solution (c) to identify the peaks due to
impurities A, Band Fj use the chromatogram obtained with
reference solution (d) to identify the peak due to impurity E.
Relative retention Widt reference to cefepime (retention
time =about 7 min): impurity E =about 0.4;
impurity F = about 0.8; impurity A = about 2.5;
A. (6R,7R)-7 -[[(2E)-(2-aminothiazol-4-yl)(methoxyimino)
acetyl] amino] -3-[(I-methylpyrrolidinio)methyl] -8-oxo-5-
thia-I-azabicyclo[4.2.0] oct-2-ene-2-carboxylate (asui-
impurity F and cefepime.
cefepime),
Limits:
impurity B =1.4; impurity E =1.8;
- impun'ty A: not more than 1.5 times the area of the
- impun'ties B, F: for each impurity, not more than the area
reference solution (b) (0.2 per cent); B. (6R,7 R)-7-[[(2Z)-[2-[[ (2Z)-(2-aminothiazol-4-yl)
- impurity E: not more than 0.5 times the area of the (methoxyimino)acetyl] amino] thiazol-4-yl] (methoxyimino)
principal peak in the chromatogram obtained with acetyl] amino J-3- [(I-methylpyrrolidinio)methyl]-8-oxo-5-
reference solution (b) (0.1 per cent): thia-I-azabicyclo [4.2.0] oct -2-ene-2-carboxylate,
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1-472 Cefi..xime 2022
PIlE" _
DEFINITION
(6R,7R)-7-[[(Zl-2-(2-Aminothiazol-4-yl)-2-
[(carboxymethoxy)imino]acetyl]amino]- 3-ethenyl-8-oxo--5-
thia-I-azabicyclo[4.2.0[oct-z-ene-2-earboxylic acid trihydrate.
Semi-synthetic product derived from a fermentation product
C. (2Zl-2-(2-aminothiazol-4-yl)-N-(formybnethyl)-2-
(methoxyimlnojacetamide, Content
CHARACTERS
Appearance
White or abnost white, slightly hygroscopic powder.
Solublllty
Slightlysoluble in water, soluble in methanol, sparingly
soluble in anhydrous ethanol, practically insoluble in ethyl
D. (2Zl-(2-aminothiazol-4-yl)(methoxyimino)acetic acid,
acetate.
0+r~~-
IDENTIFICATION
N""" N+,CH, Infrared absorption spectrophotometry (2.2.24).
H,N
H H
S 0 Comparison cefixime CRS.
If the spectra obtained show dilferences, dissolve the
substance to he examined and the reference substance
E. (6R,7KJ-7-amino-d-] (l-methylpyrrolidinio)methyl]-8-oxo- separately in methanol R, evaporate to dryness and record
5-thia-I-azabicyclo[4.2.0] oct-2-ene-2-earboxylate, new spectra using the residues.
TESTS
0+r~Co,-
N N'
,CH, pH (2.2.3)
2.6 to 4.1.
H: S 0 Suspend 0.5 g in carbon dioxide-free water R and dilute to
CH, HN~HO 10 mL with the same solvent.
N....
6 0
)-~ .. ~ N+
.... CH3 Related substances
Liquid chromatography (2.2.29). Prepare 'he solutions
H,N-{ Nn~··t-i--
I H H S 0 immediatelY before use.
S 0
F. (6R,7R)-7-[[[(6R,7R)-7-[[(2Zl-(2-aminothiazol-4- the mobile phase.
yl)(methoxyimino)acetyl]amino]-3- [(1- Reference solutwn (a) Dissolve 25.0 mg of cejixime CRS in
methylpyrrolidinio)methyl]-8-oxo--5-thia-I-azabicyclo the mobile phase and dilute to 25.0 mL with the mobile
[4.2.0] oct -2-en- 2-yl]carbonyl) amino]-3- [( 1- phase.
methylpyrrolidinio)methyl]-8-oxo--5-thia-I-azabicyclo Reference solution (b) Dilute 1.0 mL of reference solution (a)
[4.2.0] oct- 2-ene-2-earboxylate, to 100.0 mL with the mobile phase.
Reference solution (c) In order to prepare impurity D in situ,
dissolve 10 mg of cefixime CRS in 10 mL of water R, heat on
a water-bath for 45 min and cool. Inject immediately.
Column:
- size: 1= 0.125 rn, (2) = 4 mm;
G. I-methylpyrrolidine (N-methylpyrrolidine). - stationary phase: octade<y/si(yl silica gelfor chromaUJgraphy R
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<I
(5 um),
- Mobile phase: mix 250 volumes of aceronitrik Rand
750 volumes of a tetrabutylammonium hydroxide solution
Cefixime prepared as follows: dissolve 8.2 g of tetrabutylammonium
hydroxide R in waterfor chromaUJgraphy R and dilute to
800 mL with the same solvent; adjust to pH 6.5 with
dilute phosphoric acidR and dilute to 1000 mL with water
for chromaUJgraphy R.
3",0
Autosampler Set at 4 "C,
Run time 3 times the retention time of cefixime.
Action and use
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2022 Cefoperazone Sodium 1-473

cefixime and impurity D; if necessary, adjust the
concentration of acetonitrile in the mobile phase.
Limits:
- any ;mpun"ty: for each impurity) not more man 0.5 times
the area of the principal peak in the chromatogram C. (6R, 7Sj-7-[[(Z)-2-(2-aminothiazol-4-yl)-2-
obtained with reference solution (b) (0.5 per cent); [(carboxymethoxy)imino)acetyl)amino)-3-ethenyl-8-ox05-
- total: not more than 3 times the area of the principal peak thia-I-azabicyclo[4.2 .Ojoct -2-ene-2-carboxylic acid
in the chromatogram obtained with reference solution (b) (cefixime 7-epimer),
(3 per cent) j
(0.1 per cent).
Ethanol (2.4.24)
Head-space gas chromatography (2.2.28): use the standard
additions method.
Sample solution Dissolve 0.250 g of the substance to be
D. (6R, 7R)-7 -[[(E)-2-(2-aminothiazol-4-yl)-2-
examined in dimelhylfonnamide R and dilute to 25.0 mL with
[(carboxymethoxy) imino) acetyl]amino)-3-ethenyl-8-ox05-
the same solvent.
thia-I-azabicyclo[4.2.01oct-2-ene-2-carboxylic acid
Limit: (cefixime (E)-isomer),
- ethanol: maximum 1.0 per cent.
Water (2.5.12) CD,H
( CO,H
O)--~~CH3
NJrN ~ . . ~_)
,0
H,N-< I H H S
ASSAY S 0
related substances with the following modifications. E. (6R, 7R)-7 -[[(Z)-2-(2-aminothiazol-4-yl)-2-
lnj«tion Test solution and reference solution (a). [(carboxymethoxy)imino)acetyl)amino)-3-methyl-8-oxo-5-
System suitahl1ity Reference solution (a): thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
- symmetryfatter. maximum 3.0 for the peak due to
cefixime;
o Cf<,
~O)
1.0 per cent determined on 6 injections.
Calculate the percentage content of C1JII5N501S2 taking
into account the assigned content of cefixime CRS.
NJ1N' y
~ . )+ "'"
o 0
CD,H
CH,
H,N-< I H H S
STORAGE S 0
IMPURITIES F. (6R, 7R)-7 -[[(Z)-2-(2-aminothiazol-4-yl)-2-[(2-ethoxy-2-
oxoethoxy)imino] acetyl)amino] -3-ethenyl-8-oxo-5- thia-L.
azabicydo[4.2.0]oct-2-ene-2-carboxylic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph£<r
Cefoperazone Sodium
A. 2-[[(Z)-2-(2-aminothiazol-4-yl)-2- (Ph. Bur. monograph 1404)
[(carhoxymethoxy)imino)acetyl)amino]-2-[(2R)-5-methyl-
7-oxo-I,2,5,7-tetrahydro-4H-furo[3,4-d] [I ,3J thiazin-2-yl]
acetic acid,
C0 2H 0 0
~
( " " CH3
N"'
O
HN ~ H
--1NJr~~S
H,N(I H
and eplmer at C"
S 0
C"H,,;N.,NaO,S, 668 62893-20-3

B. 2- [[[(Z)-I-(2-aminothiazol-4-yl)-2-[[[(2R, 5RSj -5-methyl-
7-oxo-I,2,5,7-'etrahydro-4H-furo[3,4-d] [1,3]thiazin-2-yl) Acdon and use
methyl]amino]-2-oxoethylidene]amino]oxy]acetic acid, Cephalosporin antibacterial.
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1-474 Cefoperazone Sodium 2022
PhE" _
Flow race I mIJmin.
DEFINITION Detection Spectrophotometer at 254 run.
Sodium (6R, 7K)-7 -[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-l- Inj«tion 20 JlL of (est solution (b) and reference
yl)carbonyl] amino]-2-(-l-hydroxyphenyljacetyl]amino]-3- [[( 1- solutions (a) and (b).
methyl-IH-tetrazol-5-yl)sulfanyl] methyl]-8-oxo-5-thia-l- Run time 2.5 times me retention time of cefoperazone.
azabicyclo[4.2.010ct-2~ne-2-carboxylate. Retention time Cefoperazone = about 15 min.
Semi-synthetic product derived from a fermentation product. System suitability Reference solution (a):
Content - numberof theoretical plates: minimum 5000, calculated for
95.0 per cent to 102.0 per cent (anhydrous substance). the principal peak; if necessary, adjust me content of
CHARACTERS acetonitrile R in the mobile phase;
- symmetry faaor: maximum 1.6 for the principal peak;
Appearance
if necessary, adjust the content of acetonitrile R in the
White or slightly yellow, hygroscopic powder.
mobile phase.
Solubility
Limits:
Freely soluble in water, soluble in methanol, slightly soluble
- any impurity: for each impurity, not more than 1.5 times
in ethanol (96 per cent).
If crystalline, it shows polymorphism (5.9). obtained with reference solution (b) (l.5 per cent);
IDENTIFICATION - total: not more than 4.5 times the area of the principal
A. Infrared absorption spectrophotometry (2.2.l<f). peak in the chromatogram obtained with reference
Preparation Dissolve the substance to be examined in
methanol R and evaporate to dryness; examine the residue. - disregard limit. 0.1 times the area of the principal peak in
Comparison Ph. Bur. reference spectrum of cefoperazone sodium. (0.1 per cent).
Acetone (2.4.24, System B)
Results The principal peak in the chromatogram obtained Maximum 2.0 per cent.
with (est solution (a) is similar in retention time and size to
Sample solution Dissolve 0.500 g of the substance to be
solvent.
C. It gives reaction (a) of sodium (2.3.1).
Solvent solution Dissolve 0.350 g of aceume R in waler Rand
TESTS dilute to 100.0 mL with the same solvent. Dilute 10.0 mL of
Appearance of solution this solution to 100.0 mL with water R.
The solution is clear (2.2.1) and its absorbance (2.2.25) at Prepare each of 4 injection vials as shown in the table below:
Dissolve 2.5 g in water R and dilute to 25.0 mL with the Vial No. Sample solution Solvent solution WaierR
same solvent. (mL) (mL) (mL)
I 1.0 0 4.0
pH (2.2.3)
2 1.0 1.0 3.0
4.5 to 6.5.
3 1.0 2.0 2.0
4 1.0 3.0 1.0
Related substances Statichead-space conditions that may be used:
Liquid chromatography (2.2.29). Prepare the solucions
- equilibration time: 15 min;
immediately before use. - transfer-line temperature: 110°C.
Test solution (a) Dissolve 25.0 mg of the substance to be Temperature:
- Column: 40 °C for 10 min.
the mobile phase.
Water (2.5.12)
the mobile phase. Bacterial endotoxins (2.6.1<f)
Less than 0.20 IV/mg, if intended for use in the manufacture
Reference solution (a) Dissolve 25.0 mg of cefoperazone
dihydrace CRS in the mobile phase and dilute to 250.0 mL of parenteralJpreparations without a further appropriate
Dilute 5.0 mL of reference solution (a)
Reference solucion (b) ASSAY
to 100.0 mL with the mobile phase. Liquid chromatography (2.2.29) as described in the test for
Column: related substances with the following modifications.
- size: 1= 0.15 m, 0 = 4.6 mm; Injection Test solution (a) and reference solution (a).
- stationary phase: end-rapped IXcadecyisilyl silica gelfur System sultabi/ity Reference solution (a):
chromatography R (5 urn). - repeatability: maximum relative standard deviation of
Mobile phase Mix 884 volumes of wacer R, 110 volumes of 1.0 per cent after 6 injections.
acetonitrile R, 3.5 volumes of a 60 gIL solution of acet~ add R Calculate the percentage content of cefoperazone sodium by
and 2.5 volumes of a triethylammonium acetate solution multiplying the percentage content of cefoperazone by 1.034.
prepared as follows: dilute 14 mL of triethylamine Rand
5.7 mL of glacial acetic acid R to 100 mL with wacer R.
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2022 Ceforaxime Sodium 1-475
STORAGE
In an airtight container, proteeredfrom light, at a
temperature of 2 °C to 8°C. IT the substance is sterile, store
in a sterile, airtight, tamper-evident container.
IMPURITIES
F. (6R,7SJ-7 -[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazine-l-
yl)carbonyl] amino]-2-(4-hydroxyphenyl)aceryl] amino]-3-
[[(I-methyl-l H-telIazol-5-yl)sulfanyl]methyl]-B-oxo-5-
thia-l-azabicyclo[4.2.0] oct- 2-ene-2-carboxylic acid.
_~ PhE",
A. (5aR,6R)-6-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-l-
yl)carbonyl]amino]-2-(4-hydroxyphenyl)aceryl]amino]-
Cefotaxime Sodium ***
*** ***
5a,6-dihydro-3H, 7H-azero[2,I-b]furo[3,4-d] [1,3]thiazine-
1,7(4H)-dione,
o /CH3
~N
o
0,
_/-.J
H. NH
0
H
#~
y
CO,H
N
N'" '"'N
dr
N-- }--..
, S S N,
I H H CH,
HO' 0 477.4 64485-93-4
B. (6R,7R)-7-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-l- Action and use

yl)carbonyl]amino]-2-(4-hydroxyphenyl)aceryl]amino]-3-
[(4-methyl-5-thioxo-4,5-dihydro-1 H-IelIazol-l-yl) methyl]- Preparation
B-oxo-5-thia-I-azabicyclo[4.2.0] ocl-2-ene-2-carboxylic Cefotaxime Injection
acid, PhE", _
SH DEFINITION
N.A N-CH3 Sodium (6R,7R)-3-[(aceryloxy)methyl]-7-[[(2Z)-2-(2-
, , aminothiazol-4-yl)-2-(methoxyimino)aceryl]arnino]-B-oxo-5-
N=N
thia-l-azabicyclo[4.2.0] oct-z-ene-z-carboxylate.
Semi-synthetic product derived from a fermentation product
C. l-methyl-IH-tetrazole-5-thiol,
Content
o yCo,H
CHARACTERS
H'N··Hs N~
N '" S
Appearance
I:')
HH White or slightly yellow powder, hygroscopic.
N-NH
Solubility
Freely soluble in water, sparingly soluble in methanol.
D. (6R,7R)-7-amino-B-oxo-3-[(IH-I,2,3-triawl-4-ylsulfanyl)
methyl]-5-thia-I-azabicyclo[4.2.0] OCI-2-ene-z-cerboxyllc IDENTIFICATION
acid (7-TACA), A. Iofrared absorption spectrophotometry (2.2.24).
Comparison cefotaxime sodium CRS.
B. II gives reaction (a) of sodium (2.3.1).
TESTS
Solution S
E. (6R,7R)-3-[(aceryloxy)methyl]-7-arnino-B-oxo-5-thia-l- Appearance of solution
azabicyclo[4.2.0]ocl-2-ene-2-carboxylic acid (7-ACA), Solution S is clear (2.2.1). Add I mL of glacial acetic acidR
to 10 mL of solution S. The solution,examined immediately,
is clear.
pH (2.2.3)
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1-476 Cefotaxime Sodium 2022
Specific optical rotation (2.2.7) impurity C = about 1.9; impurity D = about 2.3;
+ 58.0 to + 64.0 (anhydrous substance). impurity F = about 2.4; impurity G = about 3.1.
Dissolve 0.100 g in wacer R and dilute to 10.0 mL with the System suitability Reference solution (c):
same solvent. - resolution: minimum 3.5 between me peaks due to
Absorbance (2.2.25) impurity E and cefotaxime;
Maximum 0.40 at 430 DID for solution S. - symmetry factor: maximum 2.0 for the peak due to
cefctaxime.
360 to 390, determined at the absorption maximum at Limits:
- impurities A, B~ C, D, E, F: for each impurity, not more
235 nm (anhydrous substance).
Dissolve 20.0 mg in waterR and dilute to 100.0 mL with the
- any otherimpurity: for each impurity, not more than
with water R. 0.2 times the area of the principal peak in me
Related substances chromatogram obtained with reference solution (b)
Liquid chromatography (2.2.29). Prepare the solutions (0.2 per cent);
immediately before use. - total: not more than 3 times the area of the principal peak
Solution A Mobile phase B, mobile phase A (14:86 VIV). in the chromatogram obtained with reference solution (b)
Test solution Dissolve 40.0 mg of the substance to be (3.0 per cent);
examined in solution A and dilute to 50.0 mL with the same - disregard lim.,r. 0.05 times the area of the principal peak in
solution. the chromatogram obtained with reference solution (b)
(0.05 per cent).
Reference solution (a) Dissolve 8.0 mg of cefotaxlme add CRS
in solution A and dilute to 10.0 mL with the same solution. Ethanol (2.4.24, System A)
100.0 mL with solution A. N,N-Dimetbylaniline (2.4.26, Method H)
Reference solution (c) Add 1.0 mL of dilute hydrochloric Maximum 20 ppm.
add R to 4.0 mL of the test solution. Heat the solution at 2-Etbylhexanoic acid (2.4.28)
40·C for 2 h. Add 5.0 mL of buffersolution pH 6.6 Rand Maximum 0.5 per cent m/m.
1.0 mL of dilute sodiwn hydroxide solution R. Water (2.5.12)
Reference solution (d) Dissolve 4 mg of cefotaxime for peok Maximum 3.0 per cent, determined on 0.300 g.
identification CRS (containing impurities A, B, C, E and F) in Bacterial endotoxins (2.6.14)
5 mL of solution A. Less than 0.05 IU/mg, if intended for use in me manufacture
Column: of parenteral preparations without a further appropriate
- size: 1= 0.15 m, 0 = 3.9 mm, procedure for the removal of bacterial endotoxins.
- stationary phase: oaadecylsilYf silica gelfor chromatography R
ASSAY
(5 urn),
- temperature: 30 "C. Liquid chromatography (2.2.29) as described in the test for
Mobile phase:
- mobile phaseA: 7.1 gIL solution of disodium hydrogen Injection Test solution and reference solution (a).
phosphate dodecahydrate R adjusted to pH 6.25 using Calculate the percentage content of Cl~1~5Na07S2by
phosphomadd R; multiplying the percentage content of cefotaxime by 1.048.
- mobile phase B: methanol R; STORAGE
In an airtight container, protected from light. If the substance
is sterile, store in a sterile, airtight, tamper-evident container.
(min) (per cent VJ1I) (per cent VIV)
• -7 .6 14 IMPURITIES
7-' 86 -> 82 14 -> 18 Specified impuriues A J B, C, D, E, F.
9 - 16 •2 I• OtherdetecMble impun·ties (the/oUowing substances 'WOuld~ if
16 - 45 82 --> 60 18 -> 40 present at a sufficient lev~ be detected by oneor otherof the tests
45 - 50 6. 4. in the monograph. They arelimited by thegeneral accepumce
50 - 55 60 -> 86 40 -> 14 cruetion for other/unspecified impurities and/or by the general
55·60 .6 t4 monograph Substances for pharmaceutical use (2034). It is
there/ore not neussary to identify these impurities for j~
Flow rate 1.0 mllmin. demonstration of compliance. See also 5.10. Control o/impurities
Detection Spectrophotometer at 235 nm. in substances for pharmaceutical use) G.
Injection 10 ~ of the test solution and reference
ldendficauon of impurities Use me chromatogram supplied
with cefotaxime for peok identification CRS and the
identify the peaks due to impurities A, B, C, E and F.
Relative retention With reference to cefctaxime (retention A. (6R,7R)-7-[[ (2Z)-2-(2-aminothiazol-4-yl)-2-
=
time about 13 min): impurity B = about 0.3; (methoxyimino)acetyl]amino]-3-methyl-8-oxo-5-thia-l-
impurity A = about 0.5; impurity E = about 0.6; azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
(deacetoxycefotaxime),
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2022 Cefoxitin Sodium 1-477
B. (6R,7R)-7 -[[(2Z)-2-(2-aminothiazol-4-yl)-2-
(methoxyimino)acetyl)amino)-3-(hydroxymethyl)-8-oxo-5-
thia-I-azabicyclo[4.2.0)oct-2-ene-2-catboxylic acid
(deacetylcefctaxime),
G. (6R,7R)- 3-[(acetyloxy)methylj-7-[[(2Z)-2-[2- [[(2Z)-2-(2-
aminothiazol-4-yl)-2-(methoxyimino)acetyl)amino)thiazol-
4-)'lj-2-( methoxyimino)acetylj aminoj-8-oxo-5-thia-l-
azabicyclo[4.2.0)oct-2-ene-2-carboxylic acid (ATA
cefotaxime).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pl>E<I
C. (6R, 7R)- 3-[(acetyloxy)methyl)-7-[[(2Z)-2-[2-

(fonn)'lamino)thiazol-4-yl)-2-(methoxyimino)acetyl)
aminoj-8-oxo-5-thia-l-azabicyclo[4.2. OJ oct- z-cnc-z- Cefoxitin Sodium
carboxylic acid (N-fonnyJcefQ[axime),
D. (6R, 7R)- 3-[(acetyloxy)methyl)-7-[[(2E)-2-(2-aminothiazol-

4-yl) -2-(methoxyimino)acetyl)aminol-s-oxo-5-thia-I> 449.4 33564-30-6
azabicyclo[4.2.0joet-2-ene-2-carboxylic acid «E)-
cefotaxime), Action and use
Preparation
Cefoxitin Injection
Pl>E<I _
DEFINITION
Sodium (6R,7S)-3-[(carbamoyloxy)methyl)-7-methoxy-8-oxo-
7-[ [(thiophen-2-yl)acetyl] amino)-5-thia-l-azabicyclo[4.2.OJ
E. (5aR,6R)-6-[[(2Z)-2-(2-aminothiazol-4-yl)-2- oct-2-ene-2-carboxylate.
(methoxyimino)acetyl)amino)-5a,6-dihydro-3H,7H-azeto
[2,I-bjfuro[3,4-d] [1,3)thiazine-I,7(4H)-dione
Semi-synl:hetic product derived from a fermentation product.
(deacerylcefotaxime lactone), Content
CHARACTERS
Appearance
White or almost white, very hygroscopic powder.
Solubility
.. Very soluble in water, sparingly solublein ethanol

(96 per cent).
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.2<f).
Comparison cefoxitin sodium CRS.
F. (6R,7R)-3-[ (acetyloxy)methylj-7 -[[(2Z)-2-[2-[[[(6R, 7R)-7-
[[(2Z) -2-(2-aminothiazol-4-yl)-2-( methoxyimino)acetyl)
aminoj-2-carboxy-8-oxo-5-thia-I-azabicyclo[4.2.0j oct-2- TESTS
en-2-yl)methyljamino)thiazol-4-ylj-2-(methoxyimino) Solution S
aceryl]aminoj-8-oxo-5-thia-I-azabicyclo[4.2.0l oct-z-cnc-z- Dissolve 2.50 g in carbon dioxide-free water R and dilute to
carboxylic acid (cefotaxime dimer), 25 mL with the same solvent.
than intensity 5 of the range of reference solutions of the
most appropriate colour (2.2.2, Me/hod II).
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1-478 Cefoxitin Sodium 2022
pH (2.2.3) Limits:
4.2 to 7.0. - impuniy I: not more than the area of the principal peak in
Dilute 2 mL of solution S to 20 mL with carbon dioxide-free the chromatogram obtained with reference solution (a)
water R. (1.0 per cent);
- impun'tks E} H: not more than 0.5 times the area of the
+ 206 to + 214 (anhydrous substance). reference solution (8) (0.5 per cent);
Dissolve 0.250 g in methanol R and dilute to 25.0 mL with - impurity J: not more than 0.3 times the area of the
the same solvent. principal peak in the chromatogram obtained with
Related substances reference solution (a) (0.3 per cent);
Liquid chromatography (2.2.29). Prepare the solutions - impuniks A J B: for each impurity, not more than
immediately before use. 0.2 times the area of the principal peak in the
SoIutwn A Dissolve 1.0 g of potassium dihydrogen chromatogram obtained with reference solution (a)
phosphate Rand 1.8 g of anhydruus disodium hydrogen (0.2 per cent);
phosphate R in 1000 mL of waterR. To 100 mL of the - unspecified imptmues: for each impurity, not more than
solution add 800 mL ofwaur R, adjust £0 pH 7.0 with twice the area of the principal peak in the chromatogram
phospho';'; acid R or a 40 WL solution of sodium Irydroxide R obtained with reference solution (b) (0.10 per cent);
and dilute to 1000 mL with waterR. - total: not more than 3 times the area of the principal peak
(3.0 per cent);
examined in solution A and dilute 10 50.0 mL with
solution A.
Reference solutWn (aJ Dilute 1.0 rnL of the test solution to (0.05 per cent).
Water (2.5./2)
Reference solutwn (b) Dilute 1.0 mL of reference solution (a)
to 20.0 mL with solution A.
Reference solutwn (c) Dissolve 5 mg of cefoxitin for peak
Less than 0.13 Hl/mg, if intended for use in the manufacture
identification CRS (containing impurities A, B, E, H, I and D
in solution A and dilute to 5 mL with solution A.
Column:
- size: 1= 0.25 ID, 0 = 4.~ nun; ASSAY
- stationary phase: phetlylsi/y/ silica gd for chromatography R Liquid chromatography (2.2.29).
(3.0 pm); Ten solution Dissolve 25.0 mg of the substance to be
- temperature: 35 QC. examined in waterR and dilute to 25.0 rnL with the same
Mobile phase: solvent.
- mobile phase A: 1.0 gIL solution of ammonium formate R Reference solution (a) Dissolve 25.0 mg of cefoxiun
adjusted to pH 2.7 with anhydrous farm;'; acid R; sodium CRS in water R and dilute to 25.0 rnL with the same
- mobile phase B: acetonitrile R; solvent.
Reference salution (b) Dissolve 20.0 mg of 2-(2-thienyl)acet;,;
Tim. MobUe phase A MobIle phase B add R in waterR and dilute to 25.0 mL with the same
(min) (per cent YIV) (per cent YIV)
solvent.
0-' 92 8
Reference solution (e) Mix 1.0 mL of reference solution (a)
5 - 50 92 -t 74 8 -t 26
and 5.0 mL of reference solution (b).
50 - 85 74 26
Column:
=
- size: 1= 0.25 m, 0 4.6 rnm,
- statwnary phase: actade<ylsilyl silica gelfar chromatography R
Detection Spectrophotometer at 254 nID. (5 urn),
Inj,dan 20 JIL. Mobile phase acetic acid R) aceumitrile R) water R
Identification of impurities Use the chromatogram supplied (1:19:81 VIVIV).
with cefOXliinfar peak identification CRS and the Flow rate 1 mUmin.
the peaks due to impurities A, BJ EJ H, I and J.
Relative retention With reference to cefoxitin (retention solutions (a) and (c).
= =
impurity I about 0.98; impurity H about 1.06; Run time 12 min.
impurity E = about 1.11; impurity B = about 1.18; System suitability Reference solution (c):
=
impurity J about 1.66. - resolution: minimum 3.5 between the 2 principal peaks.
Systemsuitability Reference solution (c): Calculate the percentage content of CloHloN,Na07S2 taking
- resolution: minimum 2.0 between the peaks due to into account the assigned content of cefoxiu"tl sodium CRS.
impurities Hand E; STORAGE
=
- peak-to-valley ratio: minimum 2.0, where Hp height In an airtight container. If the substance is sterile, store in a
above the baseline of the peak due to impurity I and sterile, airtight, tamper-evident container.
Ht> = height above the baseline of the lowest point of the
curve separating this peak from the peak due to cefoxitin. IMPURITIES
Specified impurities A, B, E, H, I,].
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2022 Cefpodoxime Proxetil 1-479
Otherdetectable impurities (thefollowing substances uould, if

present ar a sufficient level, be detected by oneor other of the tesu
monograph Substance.< for pharmaceutical we (2034). 1/ is
demonstration of compliance. See also5.10. Control of impuriues F. (6R,7S)-3-[(carbamoyloxy)methyl]-7-methoxy-7-[[(2S)-2-
in subsronces forphannaceutical use) C, D, F, G. methoxy-2-( thiophen-2-yJ)acetyl)amino]-8-oxo-5-thia-l-
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid «S)-methoxy
cefoxitin),
A. (6R,7S)-3-(hydroxymethyl)-7-methoxy-8-oxo-7-
[[(thiophen-2-yl)acetyl] amino]-5-thia-l-azabicyclo [4.2.0]
oct-2-ene-2-carboxylic acid (decarbamoylcefoxitin),
H ~H 0
H°1=}Y"
I OA NH
2
{SI0
S
\-Y ~
N··
0 H S
'CH)
endepmer et c'
G. (6R,7 S)-3-[[[[[[(6R, 7S)-2-carboxy-7-methoxy-8-oxo-7-
[[2-( thiophen-2-yl)acetyl] amino]-5-thia-l-azabicyclo[4.2.0]
oct-z-en-3-yl]methyI)oxy]carbamoyl]oxy]methyl]-7-
methoxy-8-oxo-7-[[2-( thiophen-2-yl)acetyI) ami no]-5-thia-
B. (2RS,6R, 7S)-3-[(carbamoyloxy)methyl]-7-methoxy-8-oxo-
l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (cefoxitin
7- [[(thiophen-2-yl) acetyl]amino]-5-thia-l-azabicyclo [4.2.0] dimer),
oet-3-ene-2-carboxylic acid (delta-3-cefoxitin),
H. unknown structure,
I. unknown structure,
J. unknown structure.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PllEil
C. (5aR,6R)-6-[[(thiophen-2-yl)acetyl]amino]-5a,6-dihydro-
Cefpodoxime Proxetil
3H,7H-azeto[2, I-b]furo[3,4-d] [1,3]thiazine-l,7(4H)-dione (Ph. Bur. monograph 2341)
(cefalorin lactone),
D. (5aR,6S)-6-methoxy-6-[[(thiophen-2-yl)acetyl]amino]-
5a,6-dihydro-3H, 7H-azeto[2,I-b]fuco[3,4-d] [1,3]thiazine- 557.6 87239-81-4
l,7(4ll)-dione (cefoxitin lactone),
Action and use
PIlE" _
DEFINITION
(IRS)-I-[[( l-Methylethoxy)carbonyl)oxy]ethyl (6R,7R)-7-
[[(2Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)
acetyl] amino]-3-( methoxymethyl) -8-oxo-5-thia-l-
E. (6R,7S)-3-[ (carbamoyloxy)methyl]-7-methoxy-7 -[[(2R)-2-
azabicyclo [4.2.0] oct-z-ene-z-carboxylate.
methoxy-2-(thiophen-2-yl)acetyl) amino]-8-oxo-5-thia-l-
azabicyclo[4.2.0]oc'-2-ene-2-carboxylic acid «R)-methoxy Semi-synthetic productderived from a fermentation product.
cefoxitin), Content
CHARACTERS
Appearance
White or pale yellow or light brown, amorphous powder.
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1-480 Cefpodoxime Proxetil 2022
Solubility diastereoisomer I of impurity B ;:::: about 0.68;

Veryslightly soluble or practically insoluble in water, very diastereoisomer I of cefpodoxime proxetil ;:::: about 0.74;
soluble in acetonitrile and in methanol, freely soluble in impurity C ;:::: about 0.82; dlasrereolacmer IT of
anhydrous ethanol. impurity B = about 0.85; impurity 0 (2 peaks) = about 0.88
IDENTIFICATION
and 1.13; peaksdue to diastereoiscmera of impurity H:
between about 1.9 and 2.3.
System suirabi/iry:
Comparison ufpodoximeproxetil CRS.
- the chromatogram obtainedwith reference solution (b) is
TESTS similar to the chromatogram supplied with cejpodoxime
Dlastereoisomer ratio proxeiilforpeak identification CRS;
Liquid chromatography (2.2.29) as described under Assay. - resolution: minimum 6.0 between the peaks due to
Use the normalisation procedure. cefpodoxime prcxetil diastereoisomers I and IT in the
Limit Test solution: chromatogram obtained with reference solution (a);
- the ratio of the area of the peakdue to cefpodoxime - peak-to-valley ratio: minimum 1.1, where Hp ;:::: height
proxetil diasrereoisomer II to the sum of the areas of the above the baseline of the peak due to diastereoisomer II
peaks due to cefpodoxime proxetil diaatereolsomers I of impurity Band H" ;:::: height above the baselineof the
and II is between 0.5 and 0.6. lowest point of the curve separating this peak from the
peak due to impurity C in the chromatogram obtained
Related substances
with reference solution (b).
Liquid chromatography (2.2.29). Prepare 'he solurions
immediately before use or store them at 2-8 °C. Limits:
- impuniy C: not more than twice the sum of the areas of
Solvent mixture glacial acetic acid RJ acetonitrile R, water R
the 2 principal peaks in the chromatogram obtained with
(2:99:99 VIVIV).
Testsolution Dissolve 50 rngof the substance to be - impun"ty D (sum of the 2 diasureoisomen): not more than
examined in the solvent mixture and dilute to 50.0 mL with the sum of the areas of the 2 principal peaks in the
the solvent mixture. chromatogram obtainedwith reference solution (a)
Reference solution (a) Dilute 1.0 mL of the test solution to (1.0 per cent);
100.0 mL with the solvent mixture. ~ impun·ty H (sum of the diastereoisomers): not more than the
Reference solution (b) Dissolve 5 mg of cefpodoxime proxetil for sum of the areas of the 2 principal peaks in the
peak identification CRS (containing impurities B, C and D) in chromatogram obtained with reference solution (a)
5.0 mL of the solvent mixture. (1.0 per cent);
Reference solution (c) Dissolve 5 mg of cejpadoxime pro",tilfor - impuniy B (sum cf the 2 diastereuisomers): not more than
impurity H identification CRS in 5.0 mL of the solvent 0.5 times the sum of the areas of the 2 principal peaks in
mixture. the chromatogram obtained with reference solution (a)
(0.5 per cent);
Column:
- any other impurity: for each impurity, not more than
~ size: I;:::: 0.15 m, 0 ;:::: 4.6 mm;
0.2 times the sum of the areas of the 2 principal peaks in
- srationary phase: end-capped octadecylsiiYI silica gelfor
(0.2 per cent);
~ temperature: maintain at a constant temperature of 20 "C.
- total: not more than 4 times the sum of the areas of the
MoMe phase: 2 principal peaks in the chromatogram obtained with
- mobile phase A: anhydrous formic acid R, methanol R, reference solution (a) (4.0 per cent);
water R (1:400:600 VIVIV); - disregard limit: 0.05 times the sum of the areas of the
- mobile phase B: anhydrous formic add R, waterR, 2 principal peaks in the chromatogram obtained with
merhanol R (1:50:950 VIVIV); reference solution (a) (0.05 per cent).

Water (2.5.12)
(min) (per cent VII-? (per cent VII-? Maximum 2.5 per cent, determined on 0.500 g.
0-65 9S s ASSAY
65 - 145 95 ...... 15 5 ...... 85 Liquid chromatography (2.2.29).
145 ~ 155 15 8' Solurion A 20 mgIL solution of anhydrous cirric acid R in
acetonitrile R.
Flow rare 0.6 mUmin. Test solution Dissolve 30.0 mg of the substance to be
Detection Spectrophotometer at 254 run. examined in solution A and dilute to 50.0 mL with
Injection 20 ~L. solution A.
Idendfication 0/impurities Use the chromatogram supplied Reference solution Dissolve 30.0 mg of cefpodoxime
with cefpodoxime proxeril for peak iden'ification CRS and the pro",u'l CRS in solution A and dilute to 50.0 mL with
chromatogram obtained with reference solution (b) to solution A.
identify the peaks due to impurities B, C and D; use the Column:
chromatogram supplied with "'fpodoxime pro",tilfor - size: I;:::: 0.15 01, 0;:::: 4.6 mm;
impurity H identification CRS and the chromatogram obtained - stationary phase: end-capped octadecylsi/yl silica gelfor
with reference solution (c) to identify the peaks due to chromarograplty R (5 1lJ1l);
impurity H. - temperature: 40 "C.
Relativeretention With reference to cefpodoxime proxetil Mobile phase methanol R, warer R (9: II VIV).
diastereoiscmer IT (retention time = about 58 min): FIuw rate 0.8 mUmin.
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2022 Cefpodoxime Proxetil 1-481
Detection Spectrophotometer at 240 run. H

.:v.-O'¥"0yCH,
Injection 10 1'1-. H3C I' II
Run time 1.2 times me retention time of cefpodoxirne

proxetil diastereoisorner II.
H,CO_ o)+°yO° CH,
Retention time Cefpodoxime proxetiJ NY'rN ~.... N OCH,

diastereoisomer IT = about 30 min. HzN--( H H S
System suitability Reference solution: S 0 andepimer at C·
cefpodoxime proxetil diastereoisomers I and Il. D. (IRS)-I-[[(I-methylethoxy)catbonyl)oxy]ethyl (6R,7R)-7-
Calculate the percentage content of C21H21N509S2 from the [[(2E)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetyl]
sum of the areas of the 2 peaks due to the diastereoisomers amino)-3-(methoxymethyl)-8-oxo-5-thia-l-azabicyclo
and using the declared content of cefpodoxime proxetil CRS. [4.2.0]oct-2-ene-2-catboxylate (anti-cefpodoxime
proxeril),
STORAGE
IMPURITIES
Specified impunues B, C, D, H.
Other detectable impun"tie.s (theJot/WJing substances would, if
present at a sufficient level, be'detected by one or other of the tests
in the monograph. They are /imiud by the general a«eptance
cruenon for other/unspecified impun"ties and/or by the general
monograph Substances for pharmaceutical use (2034). 1, is
demonstration of compliance. See also5.10. Control of impw;ues E. (lRS)-I-[[(I-methylethoxy)catbonyl)oxy)ethyl (6R,7R)-3-
in substances for pharmaceutical use) A, E, F, G. (acetoxymethyl)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-
(methoxyimino)acetyl] amino]-8-oxo-5-thia-l-azabicyclo
[4.2.0]oct-2-ene-2<atboxylate (ACA-analogue of
cefpodoxime proxetil),
H
.:v.-O'¥"0yCH,
H3C I· II
A. (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-
CH,
,°
0yO° CH,
(methoxyimino)acetyl)amino)-3-(methoxymethyl)-8-oxo-5- N N'O~ )+ '"

.. OCH,
thia-l-azabicyclo[4.2.0)oct-2-ene-2-catboxylic acid
(cefpodoxime),
OHC
~N--<'r
J g HHS
S andepimer at C'
F. (lRS)-I-[[(I-methylethoxy)catbonyl]oxy)ethyl (6R,7R)-7-
[[(2Z) -2- [(2-formylamino)thiazol-4-yl)-2-(methoxyimino)
acetyl] amino]-3-(methoxymethyl)-8-oxo-5-thia-l-
azabicyclo[4.2.0)oct-2-ene-2-catboxylate (N-formyl
cefpodoxime proxetil),
B. (IRS)-I-[[(l-methylethoxy)catbonyl]oxy]ethyl (6R,7R)-7-
[[(2Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetyl)
amino]-3-methyl-8-oxo-5-thia-l-azabicyclo [4.2.0) oct-2-
ene-2-carboxylate (ADeA-analogue of cefpodoxime
proxetil),
G. (lRS)-I-[[(I-methylethoxy)carbonyl)oxy]ethyl (6R,7R)-7-
[[(2Z)-[2-(2-acetylamino)thiazol-4-yl)-2-(methoxyimino)
acetyl]amino]-3-( methoxymethyl)-8-oxo-5-thia-l-
azabicyclo[4.2.0)oet-2-ene-2-catboxylate (N-acetyl-
cefpodoxirne prcxetil},
C. (IRS)-I-[[(l-methylethoxy)catbonyl]oxy)ethyl (6R,7R)-7-
[[(2Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetyl)
amino)-3-(methoxymethyl)-8-oxo-5-thia-l-azabicyclo
[4.2.0)oct-3-ene-2<atboxylate (delta-2-cefpodoxime
proxetil),
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1-482 Cefprozil Monohydrate 2022
TESTS
Related substances
Liquid chromatography (2.2.29). Prepare the solutiotlS
Teslsolution (a) Dissolve 0.125 g of the substance to be
examined in I mL of a 103 gIL solution of hydrochloric acid R
and dilute to 25.0 mL with mobile phase A.
H Testsolution (b) Dissolve 30.0 mg of the substance to be
.;v.0'y"0yCH, examined in water R and dilute to 100.0 mL with the same
\. = CH3 0yO
HJC II II
0 CH3
solvent.
~
N' HH'<::::::
° OCH 3
100.0 rnL with mobile phase A.
Reference solution (b) Dissolve 5 mg of cefproz.7Jor peak
\HN-{NYrN
I H HS identification CRS (containing impurities B, Hand M) in
S 0 0.05 rnL of a 103 gIL solution of hydrochloric acidR and add
and dlastereolscmers at Cf andCl' 1 mL of mobile phase A.
Reference solution (c) Dissolve 3 mg of cefprozll CRS and
H. mixture of the dlasrereoisomers of 1-[[(1- 6 mg of cefprozil impurity mixture CRS (containing
methylethoxy)carbonyl)oxy)ethyl (6R, 7K)-7 -[[(2Z)-2-[2- impurities D and F) in 2 mls of a 103 gILsolutionof
[[(2R)-2-[[(2Z)-2-(2-aminothiazol-4-yl)-2-(methoxyimino) hydrochloric add R and dilute to 50 rnL with mobile phase A.
acetyl) amino]-2-[(2R)-5-( methoxymethyl)-4-[[1-[[( 1-
Reference solutiotl (d) Dissolve 30.0 mg of cefproz.7 CRS in
methylethoxy)carbonyl)oxy]ethoxy]carbonyl]-3,6-dihydro-
2H-1 ,3-thiazin-2-yl)acetyl] amino) thiazol-4-yl)-2-
(methoxyimino)acetyl]amino)-3-(methoxymethyl)-8-oxo-5- Reference solution (e) Dissolve 10.0 mg of ceJadroxil CRS
thia- I -azabicyclo[4.2.0)oct-g-ene-g-carboxylate (impurity B) in waterR and dilute to 20.0 rnL with the same
(cefpodoxime proxetil dimer). solvent. Dilute 1.0 mL of the solution to 20.0 mL with
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE..
water R.
Reference solution (J) Dissolve 10.0 mg of cefprozil
impurity A CRS in water R and dilute to 100.0 mL with the
same solvent. Dilute 1.0 mL of the solution to 10.0 mL with
water R.
Cefprozil Monohydrate Column:
(ph. Eur. monograph 2342) - size: 1 = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-copped ocwd«y/silyl silica gelJor
Co.H chromawgraphy compotible with 100 percent aqueous mobile
H""~CH3
n°
phases R (5 ~);
- wnperalUre: 40 °C.
~
H . NIi, .",0
'" Mobile phase:
H H S - mobile phase A: dissolve 11.5 g of ammonium dihydrogen
HO 0 and(Z}-lsomer
phosphate R in waterfor chromatography R, adjust to
pH 4.4 with phosphone add R and dilute to 1000 rnL with
407.4 121123-17-9 waterfor chromatography R;
- mobile phase B: acetomtriie Jorchromawgraphy R, mobile
Action and use phase A (50:50 VIII);
PhE.. _ Time MobUe phase A Mobile phase B
(min) (per cent VIJI) (per cent VIJI)
DEFINITION 0-8 81 I.
Mixture of the 2 diasrereoisomers of (6R,7R)-7 -[[(2K)-2- S - 20 81 --l 36 19 --l 64
amino-2-(4-hydroxyphenyl)acetyl]amino)-8-oxo-3-[(IEZ)- 20 - 25 36 64
prop-I-enyl]-5-thia-I-azabicyclo [4.2.0)oct-2-ene-2-carboxylic
acid monohydrate, Flow rate 1.0 mUmin.
Semi-synthetic product derived from a fermentation product. Detection Spectrophotometer at 230 run.
Content InjecuOn 10 J.lL of test solution (a) and reference
96.0 per cent to 102.0 per cent (anhydrous substance). solutions (a), (b), (c), (e) and (I).
CHARACTERS Identification of impurities Use the chromatogram supplied
Appearance with cefprozl1 Jorpeak identification CRS and the
White or yellow, crystalline powder, slighdy hygroscopic. chromatogram obtained with reference solution (b) to
Solubillty identify the peaks due to impurities B, H and M; use me
Slightly soluble in water and in methanol, practically chromatogram supplied with cefprozll impurity mixture CRS
insoluble in acetone. and the chromatogram obtained with reference solution (c)
to identify the peaksdue to impurities D and F; impurities G
IDENTIFICATION and I are identified by their relative retention.
Comparison cefprozil CRS.
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2022 Cefprozil Monohydrate 1-483
Relative retention With reference to cefprozil (Z)-isomer Otherdetectable impurities {the following substances sooutd, if
(retention time = about 7 min): impurity A = about 0.4; present at a sufficient level, be detected by oneor otherof the tests
impurity B = about 0.5; impurity D = about 0.7; in the monograph. They are limited by the general acceptance
impurity F = about 0.9; cefprozil (E)-isomer = about 1.4; criterion for otherlunspedfied impun'tks and/or by the general
impurity G = about 1.7; impurity H = about 2.0; monograph Substances for pharmaceutical use (2034). 1, is
impurity I = about 2.1; impurity M = about 2.9. therefore nOI necessary to identify these impurities for
System suitability Reference solution (c): demonstration of compliance. See also 5.10. Control 0/ impurities
- resolution: minimum 1.4 between the peaksdue to in substances for pharmaceutical use) C, E, F, J, K, L, N.
impurity F and cefprozil (Z)-isomer.
Limits:
- cometion factor. for the calculation of content, multiply me
peak area of impurity D by 2.3;
- impurity B: not more than the area of the principal peak in
(0.5 per cent); A. (2R)-2-amino-2-(4-hydroxyphenyl)acetic acid (p-
- impun'ties D, G, H, I, M: for each impurity, not more than hydroxyphenylglycine),
0.3 times the sum of the areas of the 2 principal peaks in
yC02HCH
(0.3 per cent);
H, NH"
o
+r"'" a
vr
- impun'ty A: nor more than the area of the principal peak ~- -
in the chromatogram obtained with reference solution (.f)
(0.2 per cent); HO
'"
"%. 0
HH S
- any other impuniy: for each impurity, not more than

0.2 times the sum of the areas of the 2 principal peaks in B. (6R, 7 KJ-7-[[(2R)-2-amino-2-( 4-hydroxyphenyl)
the chromatogram obtained with reference solution (a) acetyl]amino]-3-methyl-8-oxo-5-thia-I-azabicydo[4.2.0]
(0.2 per cent); oct-2-ene-2-carboxylic acid (cefadroxd),
- disregard limit: 0.05 times the sum of me areas of the
o ~:(NH2
D
~
2 principal peaks in the chromatogram obtained with
reference solution (a) (0.05 per cent). and epimeralC'
'"
(E)-isomer ratio I ···f.N
H H
0
Liquid chromatography (2.2.29) as described under Assay. HO "'-
Determine the area of the peak due to the (E)-isomer in the
chromatogram obtained with test solution (b) and reference C. (6RSJ-3-(aminomethylene)-6-(4-
solution (d). Calculate the ratio of me (E)-isomer to the sum hydroxyphenyljpiperazine-z.f-dione,
of both cefprozil isomers, as determined under Assay.
Limit.
- (E)-isomer ratio: 0.06 to 0.11.
Water (2.5.12)
Maximum 0.2 per cent, determined on 1.0 g. D. (6R,7R)-7 -amin0--8-oxo-3-[(IZ)-prop-I-enyl]-5-thia-l-
azabicydo[4.2.0]oct-2-ene-2-carboxylic acid,
ASSAY
Mobile phase Mobile phase B, mobile phase A (18:82 VIV).
Detection Spectrophotometer at 280 nrn.
Injection 10 IJL oftest solution (b) and reference
solution (d).
Run time Twice the retention time of cefprozil (Z)-isomer.
Elution order (Z)-isomer, (E)-isomer. E. (6R,7R)-7 -[[(2R)-2-amino-2-[4-[[(2R)-2-amino-2-(4-
Retention lime Cefprozil (Z)-isomer = about 8 min. hydroxyphenyl)acetyl] oxy]phenyl] acetyl]amino l-a-oxo-3-
[( 1Z)-prop-I-enyl]-5-thia-I-azabicydo[4.2.0] oct- z-ece- 2-
carboxylic acid,
cefprozil (Z)-isomer and the (E)-isomer.
Calculate the percentage content of the sum of both isomers
of cefprozil (C,sH'9N30,S) taking into account the assigned
contents of both (E)-isomer and (Z)-isomer of cefprozil CRS.
STORAGE
F. (6R, 7R)-7 -amino-8-oxo-3-[(iE)-prop-l-enyl]-5-thia-l-
IMPURITIES azabicyclo[4.2.0]oet-2-ene-2-carboxylic acid,
Specified impurities A, B, D, G, H, I, M.
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1-484 Cefradine 2022
o OH~eo,H
NH'~·~S
m
H
-, ca,
I H H
HO ~ 0
G. (2R)-2-[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino- M.(6R,7R)-7-[[(2R)-2-amino-2-[4-[(ethoxycarbonyl)oxy]
2-[(2R) -4-carboxy-5-[(I Z)-prop-I-enyl) -3,6-dihydro-2H- phenyl] acetyl]amino]-8-oxo-3- [( IZ)-prop-l-enyl]-5-thia-1-
IJ3-dtiazin-2-yl]-acetic acid, azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,
co,H
o ~eH,
o 0~-#)
H
3C
»<: O)....oN J H H
N. (6R, 7R)-7-[[(2R)-2-amino-2-[4-(ethoxycarhonyl)oxy]
H. (6R, 7R)-7 -[[(2R)-2-[[(2R)-2-amino-2-(4-
phenyl] acetyl]amino]-8-oxo-3- [(1E)-prop-l-enyl]-5-thia-l-
bydroxyphenyl)acetyl]amino]-2-(4-hydroxyphenyl)acetyl]
azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid.
amino]-8-oxo-3-[(1Z)-prop-l-enyl]-5-thia-l-azabicyclo
__ ~ ~ Phf",
[4.2.0]oct-2-ene-2-carboxylic acid,
N~~ __
01toH~eH ***
or Cefradine
H.,
*** ***
HN.... ) 3
,po'l H H
S
***
HO' 0
I. (2R)-2-[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino- \_~x.::eH'
2- [(2R) -4-carboxy-5-[(I E)-prop-l-enyI]-3,6-dihydro-2H-
1,3-thiazin-2-yl]-acetic acid, RX;~ . r-t)
° H H
Compound R/ Mol.Formula M,
cefradine
o C'~19N30",S 349.4
J. (6R, 7R)-7-[[(2R)-2-[[(2R)-2-amino-2-( 4-hydroxyphenyl)

cefalexin
o C,&Hl1N30",S 347.4
acetyl] amino ]-2-(4-hydroxyp henyl)acetyl] amino]-8-oxo-3-

[(lE)-prop-I-enyl]-5-thia-I-azabicyclo [4.2.0] oct-z-ene-z-
carboxylic acid,
4',5'-dihydrocefradine
CY C1&H2I N30"S 351A
Cefradine 38821-53-3
°t\I H
,
I~H() ;~
Action and use
Cephalosporin .antibacterial.
11'-s
~
HW
Preparations
* NH H
Cefradine Capsules
HO ~ 0 and eplmers at C· Cefradine Injection
Cefradine Oral Suspension
K. mixture of 4 diastereoisomers of (3RS,6RS)-3-[(2R,5R)-5- Phf" _
ethyl-7-oxo-I,2,5,7-tetrahydro-4H-furo[3,4-d] [1,3] thiazin-
2-yl]-6,(4-hydroxyphenyl)piperazine-2,5-dione, DEFINITION
Main component (6R,7R)-7-[[(2R)-amino(cyclohexa-I,4-
H-, NH'O dienyl)acetyl] amino]-3-methyl-8-oxo-5-thia-I-azabicyclo
HO dr ,po' I
~ 0
~OH
[4.2.0]oct-2-ene-2-carhoxylic acid (cefradine).
Content
- afradine: minimum 90.0 per cent (anhydrous substance);
L. 2-hydroxyethyl (2R)-2-amino-2-(4-hydroxyphenyl)acetate,
- cefalexin: maximum 5.0 per cent (anhydrous substance);
- 4',5'-diltydrocejradine: maximum 2.0 per cent (arthydrous
substance);
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2022 Cefradine 1-485
- sum of the percentage contents of cefradine, cefa/exin and Mobile phase:

4',5'-dihydrocefradine: 96.0 per cent to 102.0 per cent - mobile phase A: 2.72 gIL solution of potassium dihydrogen
(anhydrous substance). phosphate R adjusted to pH 3.0 with dilute phosphoric
CHARACTERS acid R;
Appearance
White or slightlyyellow, hygroscopic powder.
Solubility (min) (per cent VIY) (per cent V/l?
Sparingly soluble in water, practically insoluble in ethanol 0-2.5 99.5 ---> 97 0.5 - i 3
(96 percent) and in hexane. 2.5 - 11 97 ---> 75 :) ..... 2S
IDENTIFICATION 11-13 75 ..... 60 25 ---> 40
Infrared absorption spectrophotometry (2.2.21). 13 - 16 60 40
16·19 60 ---> 20 40 ..... 80
Comparison cefradine CRS.
19 - 19.1 20 ---> 99.5 80 ..... 05
If the spectra obtained in the solid state show differences, 19.1 - 25 99.5 0.5
dissolve 30 mg of the substance to be examined and 30 mg
of the reference substance separately in 10 mL of methanol R,
evaporate to dryness at 40 °C at a pressure less than 2 kPa Flow ra" 1.0 mUmin.
and recordnew spectrausing the residues. Detection Spectrophotometer at 220 nm.
TESTS Injectian 25 pL
Solution S Identification of impurities Use the chromatogram supplied
Dissolve 2.50 g in sodium carbonate solution R and dilute to with ce/radine for peak identification CRS and the
25.0 mL with the same solvent. chromatogram obtained with reference solution (d) to
identify the peaks due to impurities C, D and E; use the
chromatogram obtained withreference solution (a) to
identify the peak due to impurity H; use the chromatogram
suspension n (2.2.1). Allow solution S to stand for 5 min.
supplied with cejradine impuniy mixtureCRS and the
The absorbance (2.2.25) of solution S measured at 450 nm is
chromatogram obtained with reference solution (e) to identify
not greater than 0.60.
the peaks due to impurities A and G.
pH (2.2.3)
Relative retention With reference to cefradine (retention
3.5 to 6.0. = =
impurity B about 0.32; impurity C about 0.53; =
10 mL with the same solvent. =
impurity D about 0.63; impurity E about 0.80; =
Specific optical rotation (2.2.7) impurity F = about 0.92; cefalexin == about 0.95; 4 /,5'-
+ 80.0 to + 90.0 (anhydrous substance). =
dihydrocefradine about 1.06; impurity G about 1.32.=
Dissolve 0.250 g in acetal< buffer solution pH 4.6 R and dilute System suitability Reference solution (b):
to 25.0 mL with the same solution. - resolution: minimum 4.0 between the peaks due to
Related substances
Liquid chromatography (2.2.29). Limits:
Testsolution Dissolve 0.300 g of the substance to be - correaion factor: for the calculation of content, multiply the
peak area of impurity B by 3.4;
examined in mobilephase A and dilute to 50.0 mL with
mobile phase A.
- impurities A, B, C~ D, E, F, G: for each impurity, not
more than 0.25 times the area of the principal peak in the
Reference solution (aJ Dissolve 3.0 mg of cyclohexa-I,4- chromatogram obtained with reference solution (c)
dienylglycine CRS (impurity B) in mobile phase A and dilute (0.25 pet cent);
to 100.0 mL with mobile phase A. - any other impun'ly: for each impurity, not more than
Reference solution (b) Dissolve 3 mg of the substance to be 0.25 times the area of the principal peak in the
examined and 3 mg of cefalexin monohydrate CRS in mobile chromatogram obtained with reference solution (c)
phase A and dilute to 25 mL with mobile phase A. (0.25 per cent);
Reference solution (c) Dilute 1.0 mL of the test solution to - total: not more than twice the area of the principal peak in
100.0 mL with mobile phase A. the chromatogram obtained with reference solution (c)
Reference solution (eV Dissolve 6 mg of cefradine for peak (2.0 per cent);
identijicatian CRS (containing impurities C, D and E) in - disregard limit: 0.05 times the area of the principal peak in
1.0 mL of mobile phase A. the chromatogram obtained with reference solution (c)
(0.05 per cent): disregard the peaks due to cefalexin and
Reference solution (e) Dissolve the contents of a vial of
4',5 ' -dihydrocefradine.
cefradine impurity mixture CRS (impurities A and G) in
1.0 mL of mobile phase A. N,N-DImethylaniline (2.4.26, Method If)
Maximum 20 ppm.
Column:
- size: 1= 0.15 m, 0 = 4.6 nun; Water (2.5.12)
- statianary phase: octadecylsilyl silica gelfor chromatography R Maximum 6.0 per cent, determined on 0.300 g.
(5 urn), Sulfated ash (2.4.11)
- temperature: 30 "C. Maximum 0.2 percent, determined on 1.0 g.
ASSAY
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1-486 Ceftazidime Pentahydrate 2022

examined in phosphate buffer solution pH 5.0 R and dilute to
100.0 mL wilh the same solution.
Reference solution (a) Dissolve 50.0 mg of cefradine CRS
(containing 4',5'-dihydrocefradine) in phosphote blfffersolution
pH 5.0 R and dilute to 100.0 mL with the same solution.
Reference solmion (b) Dissolve 5.0 mg of cefa/exin C. (6R, 7R)-7 -[[(2R)-amino(cyclohexa-I,4-dienyl)acetyl]
monohydrate CRS in phosphate buffersolution pH 5.0 Rand amino]- 3-methyl-8-oxo-5-thia-l-azabicyclo[4.2.0] oct-2-
dilute to 100.0 mL with the same solution. ene-2-carboxylic acid 5-oxide (isomer I),
Reference solution (c) Dilute 1 mL of reference solution (a) D. (6R, 7R)-7 -[[(2R)-amino(cyclohexa-l.4-<1ienyl)acetyl)
to 10 mL with phosphate blfffersolution pH 5.0 R. Mix 5 mL amino)-3-methyl-8-oxo-5 -thla-l-azabicydc [4.2.0) oct-2-
of this solutionand 5 mL of reference solution (b). ene-2-carboxylic acid 5-oxide (isomer 2»)
Column:
- size: 1 = 0.10 m, 0 = 4.6 mm; o J:::CH
H
-, NH'~_#)
o
- stationary phase: octadecylsiiy/ silica gelfor chromatography R a
(5 J1Il1).
S
UOH~
Mobile phase methanolR, phosphate buffersolUlion pH 5.0 R HH
(25:75 VIV).
FkJw rate 1.5 mUmin.
B. (6R, 7R)-7 -[[(2R)-amino(2-hydtOxyphenyl)acetyl] amino]-
3-methyl-8-oxo-5-thia-I-azabicyclo[4.2.0) oct-2-000-2-
bijection 5 pL. carboxylic acid,
Run time Twice the retention time of cefradine.
Relative retention With reference to cefradine (retention HO CH,
time = about 3 min): cefalexin = about 0.7; 4',5'-
dihydrocefradine = about 1.5.
01:;5
- resolution: minimum 4.0 between the peaks due to F. 3-hydtOxy-4-methylthiophen-2(5H)-one,
Calculate the percentage content of cefradine using the
chromatogram obtained with reference solution (a) and
taking into account the assigned content of cejradine CRS.
Calculate the percentage content of cefalexin using the
chromatogram obtained with reference solution (b) and
taking Into account the assigned content of cefafexin
monohydrate CRS. Calculate the percentage content of 4',5'- G. (6R, 7R)-7 -[(2,2-dimethylptOpanoyl)amino)-3-methyl-8-
dihydrocefradine using the chromatogram obtained with oxo-5-thia-I-azabicyclo[4.2.0) oct-2-ene-2-carboxylic acid
reference solution (b), taking into account the assigned (7-ADCA pivalamide).
content of eefalexin monohydrate CRS and multiplying the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<6
area of the peakdue to 4/,5'-dihydrocefradine by a
correction factor of 1.6.
STORAGE
Ceftazidime Pentahydrate •••
***••****
temperature of 2 DC to 8 DC.
Ceftazidime
IMPURITIES
Specified impurities A, BJ CJ D, EJ FJ G. (Ph. Bur. monograph 1405)
A. (6R,7R)-7 -amino-3-methyl-8-oxo-5-thia-I-azabicyclo
[4.2.0]oct-2-ene-2-carboxylic acid 637 78439-06-2
(7-aminodeacetoxycephalosporanic acid, 7-ADCA),
Action and use
Preparations
Ceftazidime Eye Drops
Ceftazidime for Injection
B. (2R)-amino(cyclohexa-l,4-dienyl)acetic acid (0- Ceftazidime Injection
dihydrcphenylglycine, cyclohexa-I,4-dienylglycine),
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2022 Ceftazidime Pentahydrate 1-487
PhEtr _ _ ~ _ Flow rate 1.3 mUmin.

DEFINITION Detection Spectrophotometer at 254 run.
(6R,7R)-7-[[(2Z)-2-(2-Aminothiazol-4-yl)-2-[(I-carboxy-l- Injection 10 ~L.
methylethoxy)imino]aceryl]amino]-8-oxo-3-[(Pyridin-I-ium- Relative retention With reference to ceftazidime (retention
l-yl)methyl]-5-thia-I-azabicydo[4.2.0] OCI-z-ene-2- =
time = about 8 min): impurity F about 0.4;
carboxylate pentahydrate. impurity G =about 0.8; impurity A =about 0.9;
Semi-synthetic product derived from a fermentation product. impuriry B =about La.
Content Identification of impurities Use the chromatogram supplied
95.0 per cent 10 102.0 per cent (anhydrous substance}. with ce/tazidime for peak identification CRS and the
CHARACTERS chromatogram obtained with reference solution (c) to identify
Appearance the peaks due to impurities A and G; use the chromatogram
White or almost white, crystalline powder. obtained with reference solution (b) to identify the peak due
to impurity B.
Solubillty
Slightly soluble in water and in methanol, practically System suitability Reference solution (c):
insoluble in acetone and in ethanol (96 per cent). It dissolves - resolution: minimum 4.0 between the peaks due to
in acid and alkali solutions. impurity A and cefrazidime.
Comparison ceftazidimeCRS. - impurities A, B, G: for each impurity, not more than the
TESTS area of the principal peak in the chromatogram obtained
Solution S with reference solution (a) (0.2 per cent);
Dissolve 0.25 g in carbon dioxide-free walerR and dilute to - unspecified impurities: for each impurity, not more than
50 mL with the same solvent. 0.5 times the area of the principal peak in the
Appearance of solution chromatogram obtained with reference solution (a)
Solution S is dear (2.2.1) and colourless (2.2.2, Method If). (0.10 per cent),
pH (2.2.3) - wlal: not more than 5 times the area of the principal peak
3.0 to 4.0 for solution S. in the chromatogram obtained with reference solution (a)
(1.0 per cent};
Related substances
- disregard limir. 0.25 times the area of the principal peak In
Tesl solution Suspend 0.150 g of the substance to be (0.05 per cent); disregard the peak due 10 impuriry F.
examined in 5 mL of acetonitrile R, dissolve by adding
Impurity F
water R and dilute to 100 mL with water R.
Reference solution (a) To 1.0 rnL of the test solution add immediately before use.
5.0 mL of acetonitrile R and dilute 10 100.0 mL with water R.
Phosphate buffersolution Prepare a 10 per cent VIV solution
Dilute 1.0 mL of this solution to 5.0 mL with water R.
of phosphate buffer solution pH 7.0 R4.
Reference solution (b) In order to prepare impurity B in situ,
expose 5 mL of the test solution to ultraviolet light at
examined in phosphate buffer solution and dilute to
254 run for about 24 h.
100.0 mL with the same solution.
Reference solution (a) Dissolve 1.00 g of pyridineR in
cefiazidime for peak identification CRS (containing impurities A
water R and dilute to 100.0 mL with the same solvent. Dilute
and G) in 2.0 rnL of water R.
5.0 mL of the solution to 200.0 rnL with water R. Dilute
Column: 1.0 rnL of this solution 10100.0 rnL with phosphate buffer
- size: 1= 0.25 m, 0 = 4.6 mm; solution.
- stationary phase: octade<ylsi/yl silica gelfor chromarography R
Reference solution (b) Dilute I mL of the lest solution 10
(5 um);
200 rnL with phosphate buffer solution. To I rnL of this
solution add 20 mL of reference solution (a) and dilute to
Moln7e phase: 200 rnL with phosphate buffer solution.
- mobile phase A: solution containing 3.6 gIL of disodium
Column:
hydrogen phosphate dodecahydrate R and 1.4 WL of
- size: 1= 0.25 m, 0 = 4.6 mm;
potassium dihydrogen phosphate R, adjusted to pH 3.4 with
- stationary phase: octadecylsi/yl silica gelfor chromarography R
a 10 per cent VIV solution of phosphoric add R;
(5 urn).
- mobile phase B: acetonitrile for chromawgraphy R;
Mobile phase Mix 8 volumes of a 28.8 WL solution of
Tbn. Mobile phase A Mobile phase B ammonium dihydrogen phosphate R previously adjusted to
(min) (per cent VIJI) (per cent V/J1 pH 7.0 with ammonia R, 24 volumes of acetonitrile Rand
0-4 96 -> 89 4 -> 1l 68 volumes of waterR.
4-5 8. 11 Flow ral£ 1.0 mllmin.
5·8 89 ---> B4 11 -> 16
8 - 11 84 ---> 80 16 -+ 20
11 - 15 80 ---> 50 20 ---> 50
Injection 20 I'l-.
15 - 18 50 ---> 20 50 -+ 80 Run time 10 min.
18·22 20 80
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1-488 Ceftazidime Pentahydrate 2022
System suitability Referencesolution (b):

ceftazidime and impurity F.
Limit:
- impun'ty F: not more than the area of the principal peak in
(500 ppm).
Water (2.5.12) A. (2RS,6R, 7Kj-7 -[[(2Z)-2-(2-aminothiazol-4-yl)-2-[(I-
13.0 per cent to 15.0 per cent, determined on 0.100 g. carboxy-I-methylethoxy)irnino)acetyl)amino)-8-oxo-3-
[Ipyridin -I-ium-I-yl)methyl)-5-thia-I-azabicyclo[4.2.0] oct-
3-ene-2-carboxylate (A-2-ceftazidime),
2Y-C~CH3
y+
HOC _
procedure for the removal of bacterial endotoxins. co,
ASSAY o 'N
Nn~-#
0
I
H,N-{
S
I
0
H
N
H S
'"
0
N '"
the mobile phase.

Reference solurion (a) Dissolve 25.0 mg of ceftazidime CRS in B. (6R,7R)-7-[[(2E)-2-(2-aminothiazol-4-yl)-2-[( l-carboxy-l-,
the mobile phase and dilute to 25.0 mL with the mobile methylethoxy)imino] acetyl) amino)-8-oxo-3- [(pyridin-I-
phase. ium-I-yl)methyl)-5-thia-I-azabicyclo[4.2.0]oct-2-ene-2-
Reference solution (b) Dissolve the contents of a vial of carboxylate,
ceftazidime for peak idemificadon CRS (containing impurities A
~+
and G) in 3.0 mL of the mobile phase.
o
Column:
- size: 1= 0.15 m, 0 = 4.6 nun;
- stationary phase: hexylsilyl silica gelfor chromatography R
H,NH)
H H
0
(5 urn).
Mobile phase Dissolve 4.3 g of disodium hydrogen phosphare C. (6R,7R)-7-amino-8-oxo-3-[(pyridin-I-ium-I-yl)methyl)-5-
dodecahydrare Rand 2.7 g of potassium dihydrogen phosphare R thia -f-azablcyclc [4.2.0)oct-z-ene-2-carboxylate,
in 980 mL of water R, then add 20 mL of aceumiuile R.
Flaw rale 2 mUmin.
Injurion 20 pL.
Run time 6 min.
Relative retention With reference to ceftazidime (retention
time = about 4.5 min): impurity A = about 0.7.
Sysrem suitability Reference solution (b):
impurity A and ceftazldirne. E. (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-[[2-(I ,1-
Calculate the content of ceftazidime (C22H22N601S2) taking dimethylethoxy) -I, l-dlmerhyl-z-oxoethoxy] intino] acetyl]
into account the assigned content of C22H22N607SZ in amino)-8-oxo-3-[(Pyridin-I-ium-I-yl)methyl]-5-thia-l-
cejtazidime CRS. azabicyclo [4.2.0)oct-z-ene-2-carboxylate,
o
STORAGE
sterile, airtight, tamper-evident container. " ,I
IMPURITIES
Specified impurities A, B, F, G. F. pyridine,
Other detectable impurities (thefollowing substances WQUld, if
present at a sufficient level) be deuaed by one or other oj the tests
in the monograph. They are limited by the general acapeance
criterion for other/unspeafied impun'ties and/or by the general
monograph Subslances for pharmaceutical use (2034). It is
demonstration of compliance. Set also 5.10. Control of impurities
in substances forphannaceutical use) C, E, H.
G. 2-[[[( IZ) -1-(2-aminothiazol-4-yl)-2- [(oxoethyl)amino)-2-
oxoethylidene]amino]oxy]~2-methylpropanoic acid,
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2022 Ceftazidime Pentahydrate with Sodium Carbonate 1-489
Test solution Suspend 0.150 g of the substance to be

examined in 5 mL of acetonitrile R, dissolve by adding
waler R and dilute to 100 mL with water R.
Reference solution (a) To 1.0 mL of the test solution add
5.0 mL of acetonitrile R and dilute to 100.0 mL with water R.
Dilute 1.0 mL of this solution to 5.0 mL with water R.
Reference solution (b) In order to prepare impurity B in situ,
H. (6R,7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-[(2-methoxy- expose 5 mL of the test solution to ultraviolet light at
1,I-dimethyl-2-oxoethoxy)intino]acetyl] amino]-8-oxo-3- 254 nm for about 24 h.
[(pyridin-l-ium-I-yl)methyl]-5-thia-I-azabicyclo[4 .2.0 [oct- Reference solution (c) Dissolve the contents of a vial of
2-ene-2-carboxylate. ceftaeidime for peak identi/icaiWn GRS (containing impurities A
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEll and G) in 2.0 mL of water R.
Column:
- size: I = 0.25 m, " = 4.6 mm;
- stationary phase: oaade<ylsilyl silica gd for chromatography R
Ceftazidime Pentahydrate with (5 pm);
- temperature: 40 °e.
Sodium Carbonate for Injection Mobilephase:
(ph. Eur. monograph 2344) - mabile phase A: solution containing 3.6 gIL of disodium
hydrogen phosphate dodecahydrate R and 1.4 gIL of
Action and use
potassium dihydrogen phosphate R, adjusted to pH 3.4 with
a 10 per cent VIV solution of phosphoric acid Rj
Preparations - mobile phaseB: acetonitrile for chromatography R;
Ceftazidime Eye Drops
Ceftazidime for Injection Time Mobile phose A MobUe phase B
(min) (per cent VIJ') (per cent V/J1
Ceftazidime Injection
0-' 96 ..... 89 <I -> II
PhEII ~
'-5 8. tl
5-8 89 -> 84 II -> 16
DEFINITION
8 - 11 84 -> 80 16 -> 20
Sterile mixture of Ce/tazidinte pentahydrate (1405) and
II - 15 80 ..... 50 20 ..... 50
Anhydrous sodium carbonate (0773).
15 - 18 50 -> 20 50 -> 80
18 ~ 22 20 80
Content
- ceftazidime: 93.0 per cent to 105.0 per cent (dried and Flow rate 1.3 ml./mln.
carbonate-free substance);
- sodium carbonate: 8.0 per cent to 10.0 per cent.
Injection I0 ~L.
CHARACTERS
Relative retention With reference to ceftazidime (retention
Appearance
time == about 8 min): impurity F == about 0.4;
White or pale yellow powder.
impurity G == about 0.8; impurity A = about 0.9;
Solubility =
Freely soluble in water and in methanol, practically insoluble
in acetone. with cefiazidime for peak identification CRS and the
IDENTIFICATION chromatogram obtained with reference solution (c) {Q identify
A. Examine the chromatograms obtained in the assay. the peaks due to impurities A and G; use the chromatogram
Results The principal peak in the chromatogram obtained obtained with reference solution (b) to identify the peak due
with the test solution is similar in retention time to the to impurity .B.
principal peak in the chromatogram obtained with reference System suitability Reference solution (c):
solution (a). - resolution: minimum 4.0 between the peaks due to
B. It gives the reaction of carbonates (2.3.1). impurity A and ceftazidime.
Limits:
TESTS
Solution S
- impun"tUs A, B, G: for each impurity, not more than the
Appearance of solution with reference solution (a) (0.2 per cent);
Solution S is clear (2.2.1) and its absorbance (2.2.25) at - unspecified impun"ties: for each impurity, not more than
425 nm is not greater than 0.50. 0.5 times the area of the principal peak in the
pH (2.2.3) chromatogram obtained with reference solution (a)
5.0 to 7.5 for solution S" (0.10 per cent);
Related substances - total: not more than 5 times the area of me principal peak
(1.0 per cent);
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1-490 Ceftazidime Pentahydrate with Sodium Carbonate 2022
- disregard limir. 0.25 times the area of me principal peak in - stationary phase: hexy[silyl silica gelfor chromatography R
the chromatogram obtained with reference solution (a) (5 urn).
(0.05 per cent); disregard the peak due to impurity F. Mobile phase Dissolve 4.3 g of disodium hydrogen phosphate
Impurity F dodecahydrare Rand 2.7 g of potassium dihydrogen phosphau R
Liquid chromatography (2.2.29). Prepare the solutions in 980 mL of water R, then add 20 mL of acetonitrile R.
immediarely before use. Flow rare 2 mUmin.
Phosphou buffer solution Prepare a 10 per cent VI V solution Detection Spectrophotometer at 245 nrn,
of phosphore buffer solution pH 7.0 R4. Injection 20 ~L.
Test solutum Dissolve 0.500 g of the substance to be Run time 6 min.
examined in phosphate buffer solution and dilute to
Relative retention Withreference to ceftazidirne (retention
Reference solution (a) Dissolve 1.00 g of pyridine R in
= =
time about 4.5 min): impurity A about 0.7.
water R and dilute to 100.0 mL with the same solvent. Dilute
5.0 mL of the solution to 200.0 mL with water R. Dilute
impurity A and ceftazidime.
1.0 mL of this solution to 100.0 mL with phosphate buffer
solution. Calculate the content of ceftazidime (C 22H 22N.O,S,) taking
into account the assigned content of C22H2~607S2 in
Reference solutian (b) Dilute 1.0 mL of the test solution to
cejtazidime CRS.
200.0 mL with phosphate buffer solution. To 1.0 mL of this
solution add 20.0 mL of reference solution (a) and dilute to Sodium carbonate
200.0 mL with phosphate buffer solution. Atomic absorption spectrometry (2.2.23, Method 1).
Cdumn: Caesium chloride buffer ,oIutian To 12.7 g of caesium
- size: I = 0.25 rn, 0 = 4.6 mm; chloride R add 500 mL of water Rand 86 mL of hydrochloric
- stationary phase: octade<y/silyl siJi<a gelfor chromatography R acidR and dilute to 1000.0 mL with waterR.
(5 1'111). Sodium standard solution (1000 mglL) Dissolve 3.70 g of
Mobile phase Mix 8 volumes of a 28.8 gIL solution of sodium nitrate R in waterR and dilute to 500 mL with the
ammonium dihydrogen phosphate R previously adjusted to same solvent, add 48.5 g of n;tt;c acid R and dilute to
pH 7.0 with ammonia R) 24 volumes of acetonirrile R and 1000 mL with waterR.
68 volumes of water R. Tes£ solutWn Dissolve 650.0 mg of the substance to be
Flow rare 1.0 ml1min. examined in water R and dilute to 100.0 mL with the same
Detection Spectrophotometer at 255 om. solvent. To 10.0 rnL of this solution add 5.0 mL of caesium
chloride buffer solutionand dilute to 50.0 mL with waur R.
Injection 20 ~L.
Reference soluti<m Into 4 identical flasks, each containing
Run time 10 min. 20.0 mL of caesium chloride buffer solution, introduce
Syuem suitability Reference solution (b): respectively 0 mL, 5.00 mL, 10.00 mL and 15.00 mL of
- resolutUm: minimum 7.0 between the peaks due to sodium standard solution (1000 mgIL) and dilute to
ceftazidime and impurity F. 200.0 mL with waterR.
Limit: Source Sodium hollow-cathode lamp.
- impurity F: not more than 6 times the area of the principal Wavelength 330.2 om to 330.3 om.
solution (a) (0.3 per cent). Asomisouon device Air-acetylene flame.
Calculate the percentage content of sodiumcarbonate.
Maximum 13.5 per cent, determined on 0.300 g. Dry at STORAGE
25°C at a pressure not exceeding 0.67 kPa for 4 h then heat In a sterile, airtight, tamper-evident container) protected from
the residue at 100 °C at a pressure not exceeding 0.67 kPa light and humidity.
for 3 h. LABELLING
Bacterial endotoxlns (2.6.14) The label statesthe percentage content mlm of ceftazidime.
Less than 0.10 JU/mg, if intended foruse in the manufacture IMPURITIES .
procedure for the removal of bacterial endotoxins. Specified impurities A, B, F, G.
Otherdetectable impurities (thefollowing substances wauld, if
ASSAY present at a sufficient/eveJ) be detected by one or other oj the tests
Ceftazidlme in themonograph. They are limited by the general a«ejJcame
Liquid chromatography (2.2.29). criterion for otherlunspecified impurities and/or by the general
Test solution Dissolve 25.0 mg of the substance to he monograph Substances for phannaceutical use (2034). It is
examined in the mobile phase and dlkite to 25.0 mL with therefore not necessary to iden..fy these impurities for
the mobile phase. demonstration of compliance. See also 5.10. Control of impurities
Reference solution (a) Dissolve 25.0 mg of ceftazidime CRS in in substances for phamraautica/ use) C) E, H.
phase.
ceftaeidime for peak identification CRS (containing impurities A
and G) in 3.0 mL of the mobile phase.
Column:
- size: 1= 0.15 m, 0 = 4.6 mID;
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2022 Ceftriaxone Sodium 1-491
A. (2RS,6R, 7R)-7 -[[(2Z)-2-(2-aminothiazol-4-yl)-2-[(I- H. (6R, 7R)-7 -[[(2Z)-2-(2-aminothiazol-4-yl)-2-[(2-methoxy-

carboxy-l-methylethoxy)inlUlo]acetyl]aminoj-8-oxo-3- 1, l-dirnethyl-2-oxoethoxy)inlUlo]acetyl]amino]-8-oxo-3-
[(pyridin-I-ium-I-yl)methyl]-5-thia-L-azablcyclo[4.2.0joct- [(Pyridin-I-ium-I-yl)methyl]-5-thia-I-azabicyclo[4.2.0]oct-
a-ene-z-cerboxylate (i\.-2-ceftazidime), 2-ene-2-carboxylate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PrlE<I
Ceftriaxone Sodium ***

*** ***
(ph. Eur. monograph 0991) ***
B. (6R,7R)-7 -[[(2E)-2-(2-aminothiazol-4-yl)-2-[(I-carboxy-l-
methylethoxy)irnino[acetyl] amino]-8-oxo-3- [(Pyridin-I-
ium-I-yI)methyIj-5-thia-I-azabicyclo[4.2. OJ oct-z-ene-z-
carboxylate,
o yCO;+
~N-+L'H H
0 104376-79-6
Action and use

C. (6R, 7R)-7 -amlno-a-eso-a-I(pyridin-I-ium-I-yl)methylj-5-
thia-I-azabicyclo[4.2.0joct-2-ene-2-carboxylate,
Preparation
H,C CH, Ceftriaxone Injection
o..x. CH, PrlE<I _
H'~
H':_O O~~~~OI
DEFINITION
Disodium (6R, 7R) -7-[[(2Z)-(2-aminothiazol-4-yl)
(methoxyinlUlo)acetyljaminoj-3-[[(2-methyl-6-oxido-5-oxo-
H2N--(
NJ1~-.t-t--)
H H S
"- 2,5-dihydro-l,2,4-triazin-3-yl)sulfanyl]methylj-8-oxo-5-thia-
l-azabicyclo[4.2.0joct-2-ene-2-carboxylate 3.5 hydrate.
S a
E. (6R, 7R)-7-[[(2Z)-2-(2-aminothiazol-4-yl)-2-[[2-(I, 1- Content
dlmethylethcxy)-1, l-dimethyl-2-oxoethoxyj imino j 96.0 per cent to 102.0 per cent (anhydrous substance).
acetyllamino]-8-oxo-3-[(Pyridin-I-ium-I-yl)methyl]-5- CHARACTERS
thia-I-azabicyclo[4.2.0] oct-2-ene-z-carboxylare,
Appearance
F. pyridine,
o Almost white or yellowish, slightly hygroscopic, crystalline
powder.
Solubillty
Freely soluble in water, sparingly soluble in methanol, very
slightly soluble in anhydrous ethanol.
IDENTIFICATION
Comparison cefitiaxene sodium CRS.
TESTS
Solution S
G. 2-[[[( IZ) -1-(2-aminothiazol-4-yl)-2- [(oxoethyl)amino]-2- 20.0 mL with the same solvent.
oxoethylidene]amino]oxy]-2-methylpropanoic acid, Appearance of solution
than reference solution Y j or BYj (2.2.2).
Dilute 2 rnL of solution S to 20 rnL with waw R.
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1-492 Ceftriaxone Sodium 2022
pH (2.2.3) Injection Test solution and reference solution (a).

6.0 to 8.0 for solution S. Calculate the percentage content of C18HI~8Na201SJ from
Specific optical rotation (2.2.7) the declared content of ceftriaxone sodium CRS.
-155 to -170 (anhydrous substance). STORAGE
Dissolve 0.250 g in water R and dilute to 25.0 mL with the In an airtight container protected from light. If the substance
same solvent. is sterile, store in a sterile, airtight, tamper-evident container.
Test solution Dissolve 30.0 mg of me substance co be
the mobile phase.
Reference soluciot, (aJ Dissolve 30.0 mg of ceftriaxone
the mobilephase.
Reference sohuion (b) Dissolve 5.0 mg of ceftriaxone
sodium CRS and 5.0 mg of cef....axone impurity A CRS in the A. (6R,7Kj-7 -[[(2b)-(2-aminothiazol-4-yl)(methoxyimino)
mobile phase and dilute to 100.0 mL with the mobile phase. acetyl)amino]-3- [[(2-methyl-5,6-dioxo-1,2,5.e-retrabydro-
Reference solution (c) Dilute 1.0 mL of the test solution to I ,2,4-triazin-3-yl)sulfanyl)methyl]-8-oxo-5-
100.0 mL with the mobile phase. thia-I-azabicyclo[4.2.0]oCl-2-ene-2-<:arboxylic acid «(E)-
isomer),
Column:
-
-
size: I = 0.25 ID, 0 = 4.6 mm;
stationary phase: octadecy/silyl silica gelfor chromatography R 0?J
""
CH,
I 0
~-H
(5 lUJ1).
Mobile phase Dissolve 2.0 g of tetradecy/ammonium bromide R N N'O
and 2.0 g of lelraheptylammonium bromide R in a mixture of H,N-{'r H HS
440 mL of waler R, 55 mL of 0.067 M phosphate buffer sJ g
solution pH 7.0 R, 5.0 mL of citrate buffer solution pH 5.0
prepared by dissolving 20.17 g of citric acid monohydrate R in B. (5aR,6R)-6-[[(2Z)-(2-aminothiazol-4-yl)(methoxyimino)
800 mL of water R, adjusting to pH 5.0 with strong sodium acetyl]amino]-5a,6-dihydro-3R,7 H-azelo[2, I-b] furo [3,4-d]
hydroxide solution Rand diluting to 1000.0 mL with water R, [1,3)thiazine-J,7(4H)-dione,
and 500 mL of acetonitrile R. o
NJyO
Detection Spectrophotometer at 254 om. II NH
Injection 20 I-IL of the test solution and reference HS.....-"-N ...
I
solutions (b) and (c). CH,
Run time Twice the retention time of ceftriaxone.
C. 2-methyl-3-sulfanyl-l,2-dihydro-l,2,4-triazine-5,6-dione,
ceftriaxone and impurity A.
Limits:
- otty impun"ty: not more than the area of the principal peak
in the chromatogram obtainedwith reference solution (e)
(1.0 per cent);
in the chromatogram obtained with reference solution (c) D. S-benzothiawl-2-yl (2Z)-(2-aminothiazol-4-yl)
(4.0 per cent); (methoxyimino)thioacetate,
(0.1 per cent).
N,N-Dlmethylanillne (2.4.26, Metlwd B)
Maximum 20 ppm.
2-Ethylhexanoic acid (2.4.21f)
Maximum 0.8 per cent mlm.
E. (6R, 7R)-7 -amino- 3-[[(2-methyl-5,6-<1ioxo-I,2,5,6-
Water (2.5.12)
tetrahydro-I ,2,4-triazin-3-yl)sulfanyl]methyl) -s-cxc-s-thia-
I-azabicyclo [4.2.0] oct- 2-ene-2-carboxylic acid.
Bacterial endotoxin. (2.6.14) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph Ell
ASSAY
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2022 Cefuroxime Axetil 1-493
Column:
Cefuroxime Axetil - size: 1= 0.25 m, 0 = 4"6 mm;
(ph. Bur. monograph 1300) - stationary phase: trimethy/si/yl silica gelfor chromatography R
(5~).
H 0 CH3 Mobile phase methanol R, 23 WI- solution of ammonium

H,CY. Y dihydrogen phosphore R (38:62 VIV).
CH, 0yO 00 Flow rare 1.0 mUmin.
N'O o~~' '" O)lNH,

andepimer at C' Detection Spectrophotometer at 278 nm.
Injection 20 J.tL of the test solution and reference
((r
o
~ j---}-S
H H
Identification of impuniits Use me chromatogram obtained
with reference solution (b) to identify the pair of peaks due
to impurity A and use the chromatogram obtainedwith
510.5 64544-07-6
reference solution (c) to identify the pair of peaks due to
Action and use impurity B.
Cephalosporin antibacterial. Relative retention Wilh reference to cefuroxime axetil
Preparations diastereoisomer A: cefuroxime axetil
Cefuroxime Axetil OralSuspension diastereoisomer B = about 0.9; impurity A = about 1.2;
Cefuroxime Axetil Tablets
=
impurity B 1.7 and 2.1.
PhE" ~ _ ~ resolution: minimum 1.5 between the peaksdue to
cefuroxime axetil diastereoiscmer A and impurity A.
DEFINITION
Mixture of the 2 diasrereoisomers of (IRSJ-l-(acetyloxy)ethYI Limits:
(6R,7R)- 3-[ (carbamoyloxy)methylJ-7- [[(2Z)-2-(furan-2-yl)- - impun"ty A: maximum 1.5 per cent for me swn of the pair
2(methoxyimino)acetyl] amino]-8-oxo-5-thia-l- of peaks;
azabicyclo[4.2. OJ oct-2-ene-2-<:arboxylate. - impun"cy B: maximum 1.0 per cent for the sum of the pair
of peaks;
- impurity E: maximwn 0.5 per cent;
Content - a1'O' other impun"ty: for each impurity, maximum
96.0 per cent to 102.0 per cent (anhydrous substance). 0.5 per cent;
CHARACTERS - total: maximwn 3.0 per cent;
Appearance - disregard Iimii: 0.05 times the area of the 2 principal peaks
White or almost white powder. in the chromatogram obtained with reference solution (a)
(0.05 per cent).
Solubility
Slightlysoluble in water, soluble in acetone, in ethyl acetate Dlastereoisomer ratio
and in methanol, slightly soluble in ethanol (96 per cent). Liquid chromatography (2.2.29) as described in the test for
related substances.
IDEN11FICATION
Limit Test solution:
- the ratio of the area of the peak due to cefuroxime axetiJ
Comparison cefuroxime a:atil CRS. diastereoisomer A to me sum of the areas of the peaks
B. Examine the chromatograms obtained in the assay. due to cefuroxime axetildiastereoisomers A and B is
Results The principal peaks in the chromatogram obtained between 0.48 and 0.55.
with me test solution are similar in retention time and size to Acetone (2.4.24)
the peaks due to cefuroxime axetil diastereoisorners A and B Maximum 1. J per cent.
in the chromatogram obtainedwith reference solution (d).
Water (2.5.12)
TESTS Maximum 1.5 per cent. determined on 0.400 g-
liquid chromatography (2.2.29): use the normalisation
procedure. Prepare the testsoluMn and reference solution (d)
related substances with the following modifications"
immediately before me.
Injection Test solution and reference solution (d)"
Test so/micn Dissolve 10.0 mg of the substance to be
examinedin the mobile phase and dilute to 50.0 mL with System suitability Reference solution (d):
the mobile phase. - resolution: minimum 1.5 between the peaks due to
cefuroxime axetil diastereoisomers A and B;
2.0 per cent for the sum of the peaks due to cefuroxime
Reference solution (b) In order to prepare in situ impurity A, axetil diastereoisomers A and B after 6 injections.
heat 5 mL of the test solution at 60 °C for 1 h.
Calculate me percentage content of C2oH22N40U)S from the
Reference solution (c) In order to prepare in situ impurity B, sum of the areas of the 2 diastereoisomer peaks and the
expose 5 mL of the test solution to ultraviolet light at declared content of C,oH22N,OlOS in cefuroxime axetilCRS.
254 om for 24 h.
STORAGE
Reference sokaion (d) Dissolve 10.0 mg of cefuroxime
axetil CRS in the mobile phase and dilute to 50.0 mL with
the mobile phase.
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1-494 Cefuroxime Sodium 2022
IMPURITIES
Cefuroxime Sodium ***
Specified impuriues A, B, E. *** ***
Other detectable impurities (thefollowing substances would, if (Ph. Eur. monograph 0992) ***
present at a sufficient level, be detected by one or other 0/the tests
ctitetion far other/unspecified impun"ties and/or by the general
there/ore not necessary to identify these impurities for
demonstration af compliance. See also 5. I O. Control of impurities
in substances for pharmaceutical use) C, D.
56238-63-2
Action and use

Preparations
Cefuroxime Eye Drops
CefuroximeInjection
Cefuroxime Intracameral Injection
A. l-(acetyloxy)ethyl (6R,7R)-3-[(carbamoyloxy)methyl]-7-
PhE" _
[[(2Z}-2-(furao-2-yl)-2-(methoxyimino)acetyl]amino]-8-
oxo-5 -thia-l-azabicyclo [4.2.0] oct-S-ene-z-carboxylate (~3_ DEFINITION
isomers), Sodium (6R, 7R)-3-[(carbamoyloxy)methyl]-7-[[(Z}-(furao-2-
yl)(methoxyimino)acetyl] amino]-8-oxo-5-thia-l-azabicyclo
0 °0
H 0 CHJ
H,CY. y [4.2.0]ocl-2-ene-2-earboxylale.
HCO
3 'N H
0 0!0
)--~ ~
... O.--l.NH2
and epimerat C·
Semi-syntheticproduct derived from a fermentation product.
Content
U
'orNITs
~ .
CHARACTERS
Appearance
White or almost white, slightlyhygroscopic powder.
B. (IRS)-I-(acetyloxy)ethyl (6R,7R)-3- Solubility
[(carbamoyloxy)methyl]-7- [[(2E)-2-(furao-2-yl)-2- Freelysoluble in water, very slightly soluble in ethanol
(methoxyimino) acetyl]amino]-8-oxo-5-thia-l-azabicyclo (96 per cent).
[4.2.0]oct-2-ene-2carboxylale «E)-isomers),
IDENTIFICATION
Comparison cefuroxime sodium CRS.
TESTS
Solution S
C. (6R,7R)-7 -[[(2Z}-2-(furan-2-yl)-2-(methoxyimino) Dissolve 2.0 g in carbon dioxide-free water R and dilute to
acetyl]amino]-8-oxo-3-[[[(trichloroacetyl)carbamoyl] 20.0 mL with the same solvent.
oxy]methyl]-5-thia-l-azabicyclo[4.2.0] ocl-2-ene-2- Appearance of solution
carboxylic acid, Solution S is not more opalescent thanreference
suspension II (2.2.1). The absorbaoce (2.2.25) of solution S
measured at 450 om is not greater than 0.25.
pH (2.2.3)
5.5 10 8.5.
Dilute 2 mL of solution S 10 20 mL with carbon dioxide-jree
...:. water R.
D. cefuroxime, . Specific optical rotation (2.2.7)
+ 59 to + 66 (arthydrous substaoce).
Dissolve 0.500 g in acetate buffer solution pH 4.6 R and dilute
to 25.0 mL with the same buffersolution.
Related substances
Liquid chromatography (2.2.29). Prepare the solu,iom
immediately before use or keep at 2-8 'C.
E. (5aR,6R)-6-[[(2Z}-2-(furao-2-yl)-2-(methoxyimino)acetyl] examined in water R and dilute to 25.0 mL with the same
amino ]-5a,6-dihydro-3H,7H-azelo[2,I-b]furo[3,4-d] [1,3] solvent.
thiazine-I,7(4H)-dione (descarbamoylcefuroxime lactone). Testsolulion (b) Dilute 5.0 mL of test solution (a) 10
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" 50.0 mL with water R.
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2022 Cefuroxime Sodium 1-495
Reference solution (a) Dissolve 25.0 mg of cefuroxime Lw>URlTIES

sodium CRS in water R and dilute to 25.0 mL with the same
solvent. Dilute 5.0 mL to 50.0 mL with waterR.
Reference solution (b) Place 20 mL of reference solution (a)
in a water-hath at 80°C for 15 min. Cool and inject
immediately.
Reference solu,ion (c) Dilute 1.0 mL of test solution (a) to
A. (6R,7R)-7-[[(Z)-(furan-2-yl)(methoxyimino)acetyl]amino]-
Column:
3-(hydroxymethyl)-8-oxo-5-thia-I-azabicyclo[4.2.OJ ocr-z-
- size: / = 0.125 ID, 0 = 4.6 mm;
ene-2-carboxylic acid (descarbamoylcefuroxime)
- stationary phase: hexy/silyl silica gelfor chromatography R
(5 urn).
MoMe phase Mix 1 volwne of acetoninile Rand 99 volwnes
. of an acetate buffersolution pH 3.4, prepared by dissolving
6.01 g of glacial acetic acid Rand 0.68 g of sodium acetate R
in water Rand diluting to 1000 mL with me same solvent
Detection Spectrophotometer at 273 nm. B. (6R, 7R)-3-[(acetyloxy)methyIJ-7-[[(Z)-(furan-2-yl)
Inj«tion 20 J.lL loop injector; inject test solution (a) and (methoxyimino) acetyl]aminoJ-8-oxo-5-thia-I-azabicyclo
referencesolutions (b) and (c). [4.2.0Joct-2-ene-2-carboxylic acid,
Run time 4 times the retention time of cefuroxime.
System suiuzbility Reference solution (b):
cefuroxime and impurity A.
Limits:
- impun·ty A: not more than the area of the principal peak
(1.0 per cent); C. (6R, 7R)-7-[[(Z)-(furan-2-yl)(methoxyimino)acetyIJaminoJ-
- atry other impurity: not more than the area of the principal 3-methyl-8-oxo-5-thia-I-azabicyclo[4.2. OJ oct-2-ene-2-
peak in the chromatogram obtained with reference carboxylic acid)
- weal: not more than 3 times the area of the principal peak
in the chromatogram obtained withreference solution (c)
(3.0 per cent);
the chromatogram obtained withreference solution (c)
(0.05 per cent).
N,N-Dimethylaniline (2.4.26, Method B) D. (6R, 7R)-7-[[(Z)-(furan-2-yl)(methoxyimino)acetyIJamino]-
Maximwn 20 ppm. 8-oxo-3- [[[(trichloroacetyl)carbamoyl] oxy]methyl]-5-thia-
l-azabicyclo[4.2.0J oct-2-ene-2-carboxylic acid,
2-Ethylhexanoic acid (2.4.28)
Maximum 0.5 per cent mlm,
Water (2.5.12)
Less than 0.10 IU/mgJ if intended for use in the manufacture
procedure for the removal of bacterial endotoxins. E. (6R,7R)- 3-[(carbamoyloxy)methylJ-7-[[(E)-(furan-2-yl)
ASSAY (me thoxyimino)acetylJamino]-8-oxo-5-thia-I-azabicyclo
Liquid chromatography (2.2.29) as described in the test for [4.2.0Jocl-2-ene-2-carboxylic acid (trans-cefuroxime),
related substances with the foUowing modification.
Injeaion Test solution (b) and reference solution (a).
Calculate the percentage content of cefuroxime sodium.
HCO
3 'N 0 YCO,H
H~~"-"":>.: OH
STORAGE
In an airtight container. If the substance is sterile) storein a
(/rNt1 s
sterile, airtight) tamper-evident container.

F. (6R, 7R)-7-[[(E)-(furan-2-yl)(methoxyimino)acetyl]amino]-
3-(hydroxymethyl)-8-oxo-5 -thi a-l-azabicyclo [4.2.0] oct -2-
ene-2-carboxylic acid)
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1-496 Celecoxib 2022
IDENTIFICATION
Comparison celewcib CRS.
separately in 2-propanol R, evaporate to dryness and record
G. (6R,7R) -3- [(acetyloxy)methyI]-7- [[(E)-(furan- 2-yl)
(methoxyimino) acetyl] amino J-8-oxo-5-thia-l-azabicyclo
[4.2.0]oct-2-ene-2-carboxylic acid, TESTS
Related substances
Solvent mixture waterR, methanol R2 (25:75 VIV).
Reference solulion (a) Dissolve 50.0 mg of celecoxib CRS in
H. (5aR,6R)-6-[[(Z)-(furan-2-yl)(methoxyimino) the solvent mixture and dilute to 100.0 mL with the solvent
acetyl] am ino]-5a,6-dihydro-3H, 7H-azeto[2, I-b]fum [3,4-d] mixture.
[1,3]thiazine-I,7(4H)-dione, Reference solution (b) Dissolve 3 mg of celecoxib
impurityA CRS and 3 mg of celecoxib impurity B CRS in the
solvent mixture and dilute to 50.0 mL with the solvent
mixture. Dilute 1.0 mL of the solutionto 25.0 mL with
Reference solulien (c) Dilute 1.0 mL of the test solution to
J. (Z)-(furan-2-yl)(methoxyimino)acetic acid. solution to 10.0 mL with the solventmixture"
___ ~ PIlE" Column:
- size: 1= 0.25 rn, 0 = 4.6 mm;
- Slaticnary phase: end-eapped phenylsilyl siliaz gelfor
chromatography R (5 um);
Celecoxib - temperature: 60 "C.
Mobile phase Mix 10 volumesof acetonitrile Rl, 30 volumes
(ph. Bur. monograph 2591) of methanol R2 and 60 volumes of a 2.7 gIL solution of
H,C pH 3.0 with plwspho,.~ acidR.
~ \l 00 Flow rate 1.5 mUmin.
\ \\ I,
~S-NH, Detection Spectrophotometer at 215 om.
Injection 25 J.lI... of the test solution and reference
NAvY
I solutions (b) and (c).
-N Run time 1.5 times the retention time of celecoxib.
F,C
Identification 0/ impun"ties Use the chromatogram obtained
381.4 169590-42-5 impurities A and B.
Re/au've retention With reference to celecoxib (retention
Action and use
Cycle-oxygenase (COX-2) inhibitor; analgesic; anti-
=
time about 27 min): impurity A =about 0.9;
inflammatory.
System suitauility:
Preparation - resolution: minimum 1.5 between the peaks due to
Celecoxib Capsules impurity A and celecoxib and minimum 1.8 between the
PIlE" _~ _ peaks due to celecoxib and impurity B in the
chromatogram obtained with referenoespluticn (b).
DEFINITION Cakulalion of percentage contents:
4-[5-(4-Methylphenyl)-3-(nitluoromethyl)-IH-pyrazol-l- - for all impurities) use the concentration of celecoxib in
yl]benzenesulfonamide. reference solution (c).
Content Limits:
98.0 per cent to 102.0 per cent (anhydrous substance). - impun'ty A: maximum 0.4 per cent;
CHARACTERS - unspecified impurities: for each impurity, maximum
Appearance 0.10 per cent;
White or almost white, crystalline or amorphous powder. - total: maximum 0.5 per cent;
Solubility
Practically insoluble in water, freely soluble to soluble in Water (2.5.12)
anhydrous ethanol, soluble in methylene chloride. Maximum 0.5 per cent) determined on 0.400 g.
II shows polymorphism (5.9).
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2022 Celiprolol Hydrochloride 1-497
PbE" _
Maximum 0.2 per cent, determined on 1.0 g in a platinum
DEFINITION
crucible. N-[3-Acetyl-4-[(2RS)-3-(tert-butylamino)-2-hydroxypropoxyJ
ASSAY phenyIJ-N',N'-diethylurea hydrochloride.
Liquid chromatography (2.2.29) as described in the test for Content
related substances with the following modification. 99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
Calculate the percentage content of Cl7HI~3N302S taking Appearance
into account the assigned content of celecoxib CRS. White or veryslightly yellow, crystalline powder.
IMPURITIES Solubility
Specified impurities A. Freely soluble in water and in methanol, soluble in ethanol
Otherdetectable impurities (lh' following sub,tonces would, if (96 per cent), very slightly soluble in methylene chloride.
present at a sufficient level, be detected by oneor otherof the tests It shows polymorphism (5.9).
in the monograph. They are limited by the general aCCefltanu
criterion for other/unspecified impurities andlor by the general IDENTIFICATION
monograph Substances for pharmaceutical us, (2034). I, is A. Infrared absorption spectrophotometry (2.2.24).
therefore not nece.ssary to identify these impun'lies for Comparison celiprolol hydrochloride CRS.
demonstration of romp/ianee. See also5.10. Control of impll1;ries If the spectra obtained in the solid stateshow differences,
in substances for pharmaceutical use) B. dissolve the substance to he examined and the reference
substance separately in methanol R, evaporate to dryness and
B. It gives reaction (a) of chlorides (2.3.1).
TESTS
-0.10' to + 0.10'.
A. 4-[5-(3-methylphenyl)-3-(trifluoromethyl)-1 H-pyrazol-I- same solvent.
yl]benzenesulfonamide, Related substances
immediauly before use, except reference solution (a).
examined in mobile phaseA and dilute to 20.0 mL with
mobile phase A.
Reference so/urian (a) In order to prepare impurity A insitu,
dissolve 10 mg of the substance to be examined in mobile
phase A and dilute to 2 rnL with mobile phase A and allow
to stand for 24 h.
Reference solution (b) Dissolve 10 mg of celiprolol for sy,um
suitobl7ity CRS (containing impurities Band F) in mobile
phase A and dilute to 2 rnL with mobile phase A.
B. 4- [3-(4-methylphenyl)-5-( trifluoromethyl)-IH-pyrazol-I-
Reference ,oIUIWn (c) Dilute 1.0 rnL of the test solution to
yl]benzenesulfonamide.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PbE<I
100.0 mL with mobile phase A. Dilute 1.0 mL oflhis
Column:
- size: 1= 0.15 m, 0 = 4.6 mm;
- stationary phase: end-capped octy/sifyl silica gelfor
Celiprolol Hydrochloride chromatography R (5 prn);
MoM, phase:
- mobil,phase A: mix 0.2 mL of ,rijluoroa",tic acidR,
0.6 mL of pentaftvoropropanoic acidR, 63 mL of o",toni,,;1e
for chromatography Rand 91 rnL of tetrahydrofuran R;
• HCl dilute to 1000 mL with waterfor chromatography R;
- mobile phaseB: acetonitrile for chromatography R;

(min) (per cent VIP) (per cent JIll')
416.0 5747fJ.78-7 0·50 100 ..... 80 0 ..... 20
Action and use

Beta-adrenoceptor antagonist.
Preparation
Celiprolol Tablets Injection I 0 ~L.
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1-498 Celiprolol Hydrochloride 2022

with celiprolcl for system suitability CRS and the chromatogram
to impurities Band F; use the chromatogram obtained with
reference solution (a) to identify me peak due to impurity A.
Relative retention With reference to celiprolol (retention
time ;;;; about 10 min): impurity A = about 0.3; C. N-[3-acetyl-4-[(2RS)-3-(tert-butylamino)-2-
impurity B = about 1.4; impurity F = about 1.6. hydroxypropoxy]phenyl]-N'-zer-burylurea,
- resolution: minimum 4.0 between the peaks due to o CH, (C~
E
H OH
impurities Band F. "'<:::::: O~N..............-CH3
Calculatimr of percentage contents: oII I andenanOOmer
- correction [actor: multiply the peak area of impurity A by H C »<; N,...J'-...N .#
4.0; a ) H
- for each impurity, use the concentration of celiprolol H,C

hydrochloride in reference solution (c).
Limits: D. N-[3-acetyl-4-[(2RS)-3-(diethylamino)-2-hydroxypropoxy]
- impurity A: maximum 0.10 per cent; phenyl]-N',N'-diethylurea,
0.10 per cent;
- tocal: maximum 0.3 per cent;
an oven at J05 °C for 3 h.
ASSAY
Dissolve 0.350 g underan atmosphere of nitrogen in 50 mL
of ethanol (96 percent) R and add 1.0 mL of 0.1 M
hydrochloric acid. Carry out a potentiometric titration (2.2.21J), E. N-[3-acetyl-4-[(23)-3-[[(23)-3-[2-acetyl-4-
using 0.1 M sodium hydroxide. Read the volume added [(diethylcarbamoyl)amino]phenoxy]-2-hydroxypropyl](tert-
between the 2 points of inflexion. butyl)amino]-2-hydroxypropoxy]phenyl]-N',N'-
1 mL of 0.1 M sodium hydroxide is equivalent to 41.60 mg of diethylurea,
C,oH"CIN,O•.
STORAGE
IMPURITIES
Otherdetectable impurities (the folkJwing substances would, if
present at a sufficient level, be detected by one or otherof the tests
in the monograph. They are limiwl by thegeneral acceptance F. N-(3-acetyl-4-hydroxyphenyl)-N',N'-diethylurea,
criterion for other/unspecified impurities andJor by the general
therefore not neassary to identify these impurities for
in substances for phannaceutica/ use) B, C, D, E, F, G, H, I.
G. N-(3-acetyl-4-[[(RS)-oxiranyl]methoxy]phenyl)-N' ,N'-
diethylurea,
A. 1-[5-amino-2-[(2RS)-3-(rm-butylamino)-2-
hydroxypropoxy]phenyl]ethan-I-one,
H. N-[3-acetyl-4-[(2RS)-3-bromo-2-hydroxypropoxy]phenyl]-
N',N' -dierhylurea,
B. N,N' -bis[3-acetyl-4-[(23)-3-(rm-butylamino)-2-
hydroxypropoxy]phenyl]urea,
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2022 Cellacefate 1-499
Water (2.5.12)
M.aximum 5.0 per cent, determined on 0.500 g.
Carry out the test using a mixture of 2 volumes of methylene
chloride R and 3 volumes of anhydrous ethanol R.
Sulfated ash (2.4. 14)
I. N-acetyl-N-(4-ethoxyphenyl)-N' ,N'-diethylurea. Maximum 0.1 per cent, determined on 1.0 g.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" ASSAY
Phthaloyl groups
Dissolve 1.000 gin 50 mL of a mixture of 2 volumes of
acelOne Rand 3 volumes of ethatlol (96 per <enO R.
Cellacefate1 Add about 0.1 mL of phenolphthalein solution RI and titrate
with O. I M sodium hydroxide. Carty out a blank titration.
(CeUulose AcetatePhthalate, Ph. Bur. monograph Calculate the percentage content of phrhaloyl groups (I')
0314) using the following expression:
9004-31f-0
14910(n, - n,) 179.5S
Action and use (IOO-a)(IOO-S)m (100 - S)
Enteric coating in pharmaceutical products.
a percentage content of water (see Tests);
PhE" _ m mass of the substance to be examined, in grams;
R. volumeof 0./ M sodium hydro:cUk used in the titration, in
DEFINITION millilitres;
Partly O-acetylated and o-phthalylated cellulose. RZ volume of 0.1 M sodium hydroxitk used in the blanktitration, in
millilitres;
Content S percentage content offree acid (see Tests).
- phthaloyl groups (C.H,O,; M, 149.1): 30.0 per cent to
36.0 per cent (anhydrous and acid-free substance), Acetyl groups
- acetyl groups (C,H,O; M, 43.04): 21.5 per cent to To 0.100 g add 25.0 mL of 0.1 M sodium hydroxide and heat
26.0 per cent (anhydrous and acid-free substance). on a water-bath under a reflux condenser for 30 min. Cool,
tCHARACTERS add about 0.1 mL of phenolphthalein solUlwn RI and titrate
Appearance with O. I M hydrochlori< acid. Carty out a blank titration.
White or almost white,'free-flowing powder or colourless Calculate the percentage content of acetyl groups using the
flakes, hygroscopic. following expression:
Solublllty
4305(n, - n,) 51.82S ]
Practically insoluble in water, freely soluble in acetone, (100 _ S) - 0.5772P
[(100 - a)(IOO - S)m
soluble in diethylene glycol, practically insoluble in ethanol
(96 per cent) and in methylene chloride. It dissolves in dilute
a percentage content of water (see Tests);
solutions of alkali hydroxides.• m mass of the substance to be examined, in grams;
IDENTIFICATION n. volume of 0.1 M hydrodJloric add used in the titration, in
millililreSj
Infrared absorption spectrophotometry (2.2.24). R2 volumeof 0.1 M hydrodJlon·c add used in Ihe blanktitration, in
Comparison cellulase acetate phthalate CRS. millilitresi
P percentage content of phlhaloyl groups;
TESTS S percentage content of me acid (see Tests).
Viscosity (2.2.9)
45 ml'a-s to 90 mpa-s, determined at 25 ± 0.2 "C. STORAGE
Dissolve 15 g, calculated with reference to the anhydrous In an airtight container,
substance, in 85 g of a mixture of I part by mass of water R FUNCTIONAIJTY-RELATED CHARACTERISTICS
and 249 parts by mass of acelOne R.
This section provides infonnation on characteristics that are
Free acid recognised as being relevant control parameters for one or more
Maximum 3.0 per cent, calculated as phthalic acid functions of the substance when used as an excipient (see chapter
(anhydrous substance). 5.15). Some of the characteriuics described in theFunctionality-
Shake 3.0 g for 2 h with 100 mL of a 50 per cent VIV related charaaenstia section may alsobe present in the mandatory
solution of methanol R and filter. Wash the flask and the filter part of the monograph since lhey also represent mandatory quality
with 2 quantities, each of 10 mL, of a 50 per cent VIV ainria. In such cases, a cross-reference to the tests described in the
solution of methanol R. Combine the filtrate and washings, mandatory part is included in the Functionality-related
add 0.1 mL of phenolphthalein solution RI and titrate with characteristics section. Control of the characteristics can contribute
O. I M sodium hydroxide until a faint pink colour is obtained. 10 the qualityof a medicinal product by improving the consistency
Carry out a blank titration using 120 mL of a of the manufacturing process and the per/onnance of the medicinal
50 per cent VIV solution of metband R. product during use. W'here control methods are cited, they are
I mL of O. I M sodium hydroxide is equivalent to 8.3 mg of recognised as being suitable for thepurpose} but other methods can
free acid, calculated as phthalic acid. alsobe used. Whenwer results for a particular chamaeriuic are
reported, the control method must be indicated.
The fo1lbwing charactenuics may be relevant for cellulose acetate
phthalace used asfilmformer in gastro-resistant tablets and
I 17Jil mf)tJl)graph hasundergone pharmaCQpoeia/ hamumwMn.
capsules.
Seechapttr 5.8 PharmaaJjJOeia/ harmonisacioo.
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1-500 Cellulose 2022
Viscosity stirring at reduced speed, taking care to avoid contacting the

See Tests. sides of the bowl with the powder. Continue stirring at low
Solubility of a film speed for 15 seconds after the addition and then stir at
Dissolve about 0.15 g in 1 mL of aceume R and pour onto a 18,000 revolutions per minute for exactly 2 minutes.
clear glass plate. A film is fonned. Take a piece of the film Immerse the appropriate spindle of a rotational viscometer,
aod place it in a flask containing 0.1 M hydrochloric acid. switch on after 30 seconds and after a further 30 seconds
It does not dissolve. Then place the piece of film in a flask determine the viscosity, Appendix V H, Method ill, using a
containing phosphate buffer solution pH 6.8 R. It dissolves. speed of 20 revolutions per minute (2.09 radians per
second).
Phthaloyl groups
See Assay. Loss on drying
When dried to constant weight at 105°, loses not more than
Acetyl groups
See Assay.
___ ~ PlfE<r Sulfated ash
Not more than 5.0%, Appendix IX A. Use 2 g.
ASSAY
Heat 2 g with 75 mL of anhydrous acetic tum
under a reflux
Dispersible Cellulose condenser for 2 hours, cool and carry out Method I for non-
Action and use aqueous titration, Appendix VITI A, determining the end point
Pharmaceutical excipient. potentiometrically. Each mL of 0.1M perchloric acid VS is
equivalent to 29.6 mg of carmellose sodium.
DEFINITION
STORAGE
Dispersible Cellulose is a mixture of Microcrystalline
Cellulose co-processed with Carmellose Sodiwn that readily Dispersible Cellulose should be protected from moisture.
forms a colloidal dispersion. LABELLING
Content of carmellose sodium The label states (1) the percentage w/w of Carmellose
75.0 to 125.0% wlw of the stated amount. Sodium; (2) the viscosity of a dispersion in water of a stated
percentage wlw of CanneUose Sodium.
CHARACfERISTICS
A white or off-white, coarse or fine powder; hygroscopic.
Disperses in water producing a white, opaque dispersion or
gel; practically insoluble in organic solvents and in dilute Microcrystalline Cellulose1
acids.
IDENTIFICATION
A. Mix 6 g with 300 mL ofwaterstirring at HO
18,000 revolutions per minute for 5 minutes. A white,
opaque dispersion is obtained which does not produce a
supernatant liquid.
B. Add severaldrops of the dispersion obtained in test A to a
10% wlv solution of aluminium chloride. Each drop forms a
white, opaque globule which does not disperse on standing.
C. Add 2 mL of iodinated potassium iodide sohuion to the
dispersion obtained in test A. No blue or purplish colour is
produced.
C6l.HIOI'I +20 SI'I + 1
D. To the residue obtained in the test for Sulfated ash add
Cellulose 9004-34-6
1 mL of hydrochloric acid, evaporate to dryness on a water
bath and dissolve the residue in 20 mL of water. Action and use
The resulting solution yields the reactions characteristic of Excipient,
sodium salts, Appendix VI, except that in test A the white
PlfE<r _
precipitate produced may not he dense.
TESTS DEFINITION
Acidity or alkalinity Purified, partly depolymerised cellulose prepared by treating
pH of the dispersion obtained in me test for Apparent alpha-cellulose, obtained as a pulp from fibrous plant
viscosity, 6.0 to 8.0, Appendix V L material, with mineral acids.
Solubility tCHARACTERS
Add 50 mg to 10 mL of ammoniacal solution of copper Appearance
teuammine and shake. It dissolves completely leaving no White or almost white, fine or granular, slightly hygroscopic
residue. powder.
Apparent viscosity Solubility
60 to 140% of the declared value when determined by the Practically insoluble in water, in acetone, in anhydrous
following method. Calculate the quantity (x g) needed to ethanol, in toluene, in dilute acids and in a 50 gIL solution of
prepare exactly 600 g of a dispersion of the stated percentage sodium hydroxide.•
w/w, with reference to the dried substance. To (600-;~) g
ofwater at 23° to 25° contained in a 1000 mL high-speed I This monograph has undergcme pharmacopoeial hanmmisaricm.
blender bowl add x g of the substance being examined, See chapter5.8 Phannaropoeial'.anllonisarion.
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2022 Cellulose 1-501
Table 0316.-1. - Intrinsic viscosiry table

IntrinsIc viscosity Irtle at different values of relative viscosity '1""
[01<
0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
0.00 0.01
0""
0.115 0,125 0.134 0.143 0.152 0.161 0.170 0.180

I.I 0.098 0.106
0.207 0.216 0.225 0.233 0.242 0.250 0.259 0.268
1.2 0.189 0.198
0.293 0.302 0.310 0.318 0.326 0.334 0.342 0.350
1.3 0.276 0.285
0.391 0.399 0.0107 00414 0.422 0.,130
1.4 0.358 0.367 0.375 0.383
0.453 0.460 0.468 0.476 0.484 0.491 0.499 0507
L5 0.'137 0.445
0.522 0.529 0.536 0.544 0.551 0.558 0.566 0.573 0.580
1.6 0.515
0.602 0.608 0.615 0.622 0.629 0.636 0.642 0.649
1.7 0.587 0595
0.670 0.677 0.683 0.690 0.697 0.104 0.710 0.717
[.8 0.656 0.663
0.736 0.743 0.749 0.756 0.762 0.769 0.775 0.782
I.' 0.723 0.730
0.802 0.809 0.815 0.821 0.827 0.833 0.840 0.846

2.0 0.788 0.795
0.864 0.870 0.876. 0.882 0.888 0.894 0.900 0.906
2.1 0.852 0.858
0.924 0.929 0.935 0.941 0.948 0.953 0.959 0.965
2.2 0.912 0.918
0.983 0.988 0.994 1.000 1.006 1.011 1.017 1.022
2.3 0.971 0.976
1.033 1.039 1.044 1.050 1.056 1.061 1.067 1.072 1.078
2.4 1.028
1.094 1.100 1.105 1.111 1.116 1.121 1.126 1.131
2.5 1.083 1.089
1.147 1.153 1.158 1.163 1.169 1.174 1.179 1.184
2.6 1.137 1.142
1.200 1.205 1.210 1.215 1.220 1.225 1.230 1.235
2.7 1.190 1.195
1.250 1.255 1.260 1.265 1.270 1.275 1.280 J.285
2.8 1.240 1.245
1.295 1.300 1.305 1.310 1.314 1.319 1.324 1329 1.333
2.' 1.290
1.348 1.352 1.357 1.362 1.367 1.371 1.376 1.381

3.0 1.338 1343
1.395 1.'100 1.405 1.409 1.414 1.418 1.423 1.427
3.[ 1.386 1390
1.441 1.446 1.450 1.455 1.459 1.464 1.468 1.473
3.2 1.432 1.436
1.482 1.486 1.491 1.496 1.500 15M 1508 1.513 1517
3.3 1,477
1.525 1.529 1.533 1.537 1.542 1.546 1.550 1554 1558
3.4 1.521
1.570 1.575 1.579 1.583 1.587 1.591 1.595 1.600
3.5 1.562 1.566
1.612 1.617 1.621 1.625 1.629 1.633 1.637 1.642
3.6 1.604 1,608
1.654 1.658 1.662 1.666 1.671 1.675 1.679 1.683
3.7 1.646 1.650
1.695 1.700 1.704 1.708 1.712 1.715 1.719 1.723
3.8 1.687 1.691
1.735 1.739 1.742 1.746 1.750 1.754 1.758 1.762
3.' 1.727 1.731
1.773 1.777 l.781 1.785 1.789 1.792 1.796 1.800

4.0 1.765 1.769
1.811 1.815 1.819 1.822 1.826 1.830 1.833 1.837
4.[ 1.804 1.808
1.848 1.852 1.856 1.859 1.863 1.867 1.870 1.874
4.2 1.841 1.845
... 1.882 J.885 1.889 1.893 1.896 1.900 1.904 1.907 1.91J
4.3 1.878
1.918 J.921 1.925 1.929 1.932 1.936 1.939 1.943 1.946
1.914
1.961 1.964 1.968 1.971 1.975 1.979 1.982
4.5 1.950 1.954 1.957
1.993 1.996 2.000 2.003 2.007 2.010 2.013 2.017
4.6 1.986 1.989
2.027 2.030 2.033 2.037 2.040 2.043 2.047 2.050
4.7 2.020 2.023
2.060 2.063 2.067 2.070 2.073 2.077 2.080 2.083
4.8 2.053 2.057
2.093 2.097 2.100 2.103 2.107 2.110 2.113 2.116
4.' 2.087 2.090
2.129 2.132 2.135 2.139 2.142 2.145 2.148

5.0 2.119 2.122 2.125
2.160 2.16<1 2.167 2.170 2.173 2.176 2.180
5.1 2,151 2.154 2.158
2.192 2.195 2.197 2.200 2.203 2.206 2.209
5.2 2.183 2.186 2.190
2.221 2.224 2.227 2.230 2.233 2.236 2.240
5.3 2.212 2.215 2.218
2.252 2.255 2.258 2.261 2.264 2.267 2.270
5.4 2.243 2.246 2.249
2.282 2.285 2.288 2.291 2.294 2.297 2.300
5.5 2.273 2.276 2.279
2.312 2.315 2.318 2.320 2.324 2.326 2.329
5.6 2303 2.306 2,309
2.341 2.344 2.347 2.350 2.353 2.355 2.358
5.7 2.332 2.335 2338
2.370 2.373 2.376 2.379 2.382 2.384 2.387
5.8 2.361 2.364 2.367
2.403 2.405 2,408 2.411 2,414 2,417
5.' 2.390 2.393 2.396 2.400
2,428 2.431 2.433 2.436 2.439 2.442 2.444

6.0 2,419 2.422 2.425
2.456 2.458 2.461 2.464 2.467 2.470 2.472
6.[ 2.447 2.450 2,453
2.483 2.486 2.489 2,492 2.494 2.497 2500
6.2 2.475 2,478 2.481
2.511 2.513 2.516 2.518 2.521 2524 2.526
6.3 2503 2.505 2.508
2.537 2540 2542 2.545 2.547 2.550 2.553
6.4 2529 2532 2.534
2.563 2.566 2.568 2.571 2.574 2.576 2.579
6.5 2.555 2.558 2.561
2.590 2.592 2.595 2.597 2.600 2.603 2.605
6.6 2.581 2.584 2.587
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Intrinsic viscosity (Ill, at different values of relative viscosity 'Ird

[oJo
o~, 0.00 0.01 0.02 D.OJ 0.04 0.05 0.06 0.07 0.08 0.09
6.7 2.608 2.610 2.613 2.615 2.618 2.620 2.623 2.625 2.627 2.630
6.8 2.633 2.635 2.631 2.640 2.643 2.645 2.648 2.650 2.653 2.655
'.9 2.658 2.660 2.663 2.665 2.668 2.670 2.673 2.675 2.678 2.680
7.0 2.683 2.685 2.687 2.690 2.693 2.695 2.698 2.700 2.702 2.705
7.[ 2.707 2.710 2.712 2.714 2.717 2.719 2.721 2.724 2.726 2.729
7.2 2.731 2.733 2.736 2.738 2.740 2.743 2.745 2.748 2.750 2.752
7.3 2.755 2.757 2.760 2.762 2.764 2.767 2.769 2.711 2.174 2.776
7.4 2.719 2.781 2.783 2.786 2.788 2.790 2.793 2.795 2.798 2.800
75 2.802 2.805 2.807 2.809 2.812 2.814 2.816 2.819 2.821 2.823
7.6 2.826 2.828 2.830 2.833 2.835 2.837 2.840 2.842 2.844 2.847
7.7 2.849 2.851 2.854 2.856 2.858 2.860 2.863 2.865 2.868 2.870
7.' 2.873 2.875 2.877 2.879 2.881 2.884 2.887 2.889 2.891 2.893
7.9 2.895 2.898 2.900 2.902 2.905 2.907 2.909 2.911 2.913 2.915
'.0 2.918 2.920 2.922 2,924 2.926 2.928 2.931 2.933 2.935 2.937
'.1 2.939 2.942 2.944 2.946 2.948 2.950 2.952 2.955 2.957 2.959
'.2 2.961 2.963 2.966 2.968 2.970 2.972 2.974 2.976 2.979 2.981
'.3 2.983 2.985 2.987 2.990 2.992 2.994 2.996 2.998 3.000 3.002
'.4 3.004 3.006 3.008 3.010 3.012 3.015 3.017 3.019 3.021 3.023
'.5 3.025
3.046
3.027
3.048
3.029
3.050
3.031
3.052
3.033
3.054
3.035
3.056
3.037
3.058
3.040
3.060
3.042
3.062
3.044
3.064
'.6
'.7 3.067 3.069 3.071 3.073 3.075 3.077 3.079 3.081 3.083 3.085
as 3.087 3.089 3.092 3.094 3.096 3.098 3.100 3.102 3.104 3.106
'.9 3.108 3.110 3.112 3.114 3.116 3.118 3.120 3.122 3.124 3.126
9.0 3.128 3.130 3.132 3.134 3.136 3.138 3.140 3.142 3.144 3.146
9.1 3.148 3.150 3.152 3.154 3.156 3.158 3.160 3.162 3.164 3.166
9.2 3.168 3.170 3.172 3.174 3.176 3.178 3.l80 3.182 3.184 3.186
9.3 3.188 3.190 3.192 3.194 3.196 3.198 3.200 3.202 3.204 3.206
'.4 3.208 3.210 3.212 3.214 3.215 3.217 3.219 3.221 3.223 3.225
9.' 3.227 3.229 3.231 3.233 3.235 3.237 3.239 3.241 3.242 3.244
9.' 3.246 3.248 3.250 3.252 3.254 3.256 3.258 3.260 3.262 3.264
9.7 3.266 3.268 3.269 3.271 3.273 3.275 3.277 3.279 3.281 3.283
9.' 3.285 3.287 3.289 3.291 3.293 3.295 3.297 3.298 3,300 3.302
9.9 3.304 3.305 3.307 3.309 3.311 3.313 3.316 3.318 3.320 3.321
IntrinsIc viscosity 1'lle at dlfferent values of relative viscosity 'lr...

[oJo
0.0 0.1 0.2 0.3 0.4 0.5 0.• 0.7 0.' 0.9
"'"'
[0 3.32 3.34 3.36 3.37 3.39 3.41 3.43 3.'15 3,46 3048
11 3.50 3.52 3.53 3.55 3.56 3.58 3.60 3.61 3.63 3.64
12 3.66 3.68 3.69 3.71 3.72 3.74 3.76 3.77 3.79 J.80
13 3.80 3.83 3.85 3.86 3.88 3.89 3.90 3.92 3.93 3.95
[4 3.96 3.97 3.99 4.00 4.02 4.03 4.04 4.06 4.07 4.09
[5 4.10 4.11 4.13 4.14 4.l5 4.17 4.18 4.19 4.20 4.22
16 4.23 4.24 4.25 4.27 4.28 4.29 4.30 4.31 4.33 4.34
17 4.35 4.36 4.37 4.38 4.39 4.41 4.42 4,43 4,44 4.45
I'[9 4,46
4.57
4,47
4.58
4,48
4.59
4,49
4.60
4.50
4.61
4.52
4.62
4.53
4.63
4.54
4.64
4.55
4.65
4.56
4.66
IDENTIFICATION C. The degree of polymerisation is not more than 350.

A. Infrared absorption spectrophotometry (2.2.24). Transfer 1.300 g to a 125 mL conical flask. Add 25.0 mL of
Comparison microcrystalline cellulose CRS. water Rand 25.0 mL of cupn'ethylenediamine hydroxide
Disregard any band between 800 em" and 825 em-lor solution R. Immediately purge me solution with nitrogen R,
between 950 ern" and 1000 em-I. insert the stopper and shake until completely dissolved.
Transfer an appropriate volume of the solution to a suitable
B. Place about 10 mg on a watch-glass and disperse in 2 mL
capillary viscometer (2.2.9). Equilibrate the solution at
of iodinated zinc chloride solution R. The substance becomes
25 ± 0.1 °C for at least 5 min. Record the flow time (er) in
violet-blue.
seconds between the 2 marks on the viscometer. Calculate
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2022 Cellulose 1-503
the kinematic viscosity (VI) of me solution using the following charring. Dry at 105 °C for 1 h, allow to cool in a desiccator
expression: and weigh. Carry out a blank determination.
kl viscometer constant. an oven at 105 °C for 3 h.
Sulfated ash (1.4./4)
Dilute a suitable volume of cupriethylenediamine hydroxide Maximum 0.1 per cent, determined on 1.0 g.
solution R with an equal volume of water R and measure the Microbial contamination
flow time (tz) using a suitable capillary viscometer. Calculate TAMC: acceptance criterion 10' CFU/g (1.6./1).
the kinematic viscosity (V2) of the solvent using the following
TYMC: acceptance criterion 102 CFU/g (1.6./1).
expression:
Absence of Escherichia co/i (1.6./3).
Absence of Pseudomonas aeruginosa (1.6./3).
~ viscometer constant. Absence of StaphylOC()",", aureus (1.6./3).
Absence of So/monella (1.6./3).
Determine the relative viscosity ('1~ of me substance to be OFUNCTIONALITY-RELATED CHARACTERISTICS
examined using the following expression: This seaion provides infontlalien on charaaeriuia that are
recognised as being rdeoam contwl parameters for one or more
Vtl V2
functions of the substance when used as an exa"pient (see chapter
Determine the intrinsic viscosity (['1lJ by interpolation, using 5. /5). Some of the characteristics described in the Functiono/ity-
the intrinsic viscosity table (Table 0316.-1). related characteristics section may also bepresent in the mandatory
Calculate the degree of pclymerisation (P) using the part of the monograph since they alsorepresenl mandatory quality
following expression: criteria: In such cases, a cross-reference to the tests described in the
mandatorypan is included in the Functionality-related
characteristics section. Control of the charaaetiuia ron contribute
m[(100 - b)/100] to the quality of a medicinal product by improving the consistency
of the manafactuting process and the perfonnance of the medicinal
m mass or the substance (0 be examined. in grams; produa during use. Where ccntrol methods are cited, they are
b loss on drying, in per cent. recognised as being suitable for the purpose, but othermethods can
also be used. Whewver results for a particular characteristic are
TESTS reported, the control methodmust be indicated.
OSolubility ThefolWwing characteristics may be relevant for mi<rocrystalline
Dissolve 50 mg in 10 mL of ammoniacalsolution of copper cellulose usedas binder, diluent or disintegrant.
teoammine R. It dissolves completely, leaving no residue.e
Los. on drying
pH (1.2.3) (see Tests).
5.0 to 7.5 for the supernatant, Particle-size distribution (1.9.3/ or 1.9.38)
Shake 5 g with 40 mL of carbon dioxide-free water R for
20 min and centrifuge.
o
Conductivity (1.1.38) _______ ~ PI>E<r
The conductivity of the test solution does not exceed the
conductivity of the water by more than 75 IJS.cm-I.
Use as test solution the supernatant obtained in the test for
Microcrystalline Cellulose and ***
** **
pH. Measure the conductivity of the supernatant after a
stable reading has been obtained and measure the Carmellose Sodium
conductivity of the water used to prepare the test solution.
*****
Ether-soluble substances
Maximum 0.05 per cent (5.0 mg) for the difference between Action and use
the mass of me residue and the mass obtained from a blank. Excipiem.:
determination. Pl>EII _
Place 10.0 g in a chromatography column about 20 mm in
internal diameter and pass 50 mL of peroxide-free ether R DEFINITION
through the column. Evaporate the eluate to dryness in a Colloid-forming, powdered mixture of i'ylidocrystalline
previously dried and tared evaporating dish, with the aid of a ceJluwse (03/6) with 5 per cent 10 22 per cent of CanneHose
current of air in a fume cupboard. After all ether has sodium (0471).
evaporated, dry me residue at 105 °C for 30 min, allow to Content
cool in a desiccator and weigh. Carry out a blank 75.0 per cent to 125.0 per cent of the nominal content of
determination. . cannellose sodium (dried substance).
Water-soluble substances CHARACTERS
Maximum 0.25 per cent (12.5 mg) for the difference Appearance
between the mass of the residue and the mass obtained from White or off-white, coarse or fine, hygroscopic powder.
a blank determination. Solubility
Shake 5.0 g with 80 mL of waterR for 10 min. Filter Dispersible in water producing a white, opaque colloidal
through a fiJter paper with the aid of vacuum into a tared dispersion; practically insoluble in organic solvents and in
flask. Evaporate to dryness on a water-bath avoiding dilute acids.
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IDENTIFICATION addition of the powder and then stir at 18 000 r/min for
A. Mix 6 g with 300 mL of water R and stir at 18 000 rfmin exactly 2 min.
for 5 min. A white opaque dispersion is obtained which does Determine the viscosity with a suitable relative rotational
not produce a 'supernatant. viscometer under the following conditions:
B. Add several drops of the dispersion obtained in - spindle: as appropriate,
identification A to a 100 gIL solution of aluminium chloride R. - speed: 20 dmin.
Each drop forms a white, opaque globule which does not Immerse the spindle into the suspension immediately after
disperse on standing. preparation, switch on the rotation spindle after 30 s; after a
C. Add 2 mL of iodinated potassium iodide. solution R to the further 30 s, take scale readings and calculate the viscosity
dispersion obtained in test A. No blue or purplish colour is according to the viscometer manual.
produced. ~ PhE"
D. It complies with the limits of the assay.
TESTS
Solubllity
Powdered Cellulose1 ****
Add 50 mg to 10 mL of ammoniacalsolution of copper
tetrammine R and shake. It dissolves completely, leaving no
** *
residue.
(Ph. Bur. monograph 0315) *****
pH (2.2.3)
6.0 to 8.0 for the dispersion obtained in identification A.
an oven at 105°C.
OH
ASSAY
Heat 2.00 g with 75 mL of anhydrous acetic add R under a
reflux condenser for 2 h) cool and titrate with 0.1 kI C~lOn+205"+1
perchloric add) determining the end-point potentiometrically
Cellulose 9004-34-6
(2.2.20).
I mL of 0.1 M perchloric acid is equivalent to 29.6 mg of Action and use
carmellose sodium. Excipient.
LABELLING PhE" _
The label states the nominal content of carmellose sodium
in per cent mlm. DEFINITION
Purified, mechanically disintegrated cellulose prepared by
FUNCTIONALITY-RELATED CHARACTERISTICS processing alpha-cellulose obtained as a pulp from fibrous
This secuon provides infol111ation on characteristics that are plant material.
recognised as being relevant control parameters for one or more
junctionsof the substance when used as an excipient (see chapter tCHARACTERS
5.15). Some of the characteristics descnbed in the Functionahiy- Appearance
related characteristics section may also bepresent in the mandatory While or almost white, fine or granular powder.
part of the monograph since they alsorepresent mandatoryquality Solubllity
cmena. In such cases, a cross-reference ro the tests described in the Practically insoluble in water, slightly soluble in a 50 gIL
mandatory part is included in the Funaionality-related solution of sodium hydroxide, practically insoluble in
characteristics section. Control of the characteristics can contribute acetone, in anhydrous ethanol, in toluene, in dilute acids and
to the quality of a medi<inal product by improving the consistency in most organic solvents.•
of the mamifacmringprows and the peformonce of the medicinal IDENTIFICATION
product during use. Where control methods are cited) they are
A. Place about 10 mg on a watch-glass and disperse in 2 mL
recognised as being suitable for the purpose, but othermethods can
of iodinated zinc chloride solution R. The substance becomes
alsobe used. When?1JtT results for a para·allar characteristic are
violet-blue.
reported, the control methodmust be indicated.
B. The degree of polymerisation is greater than 440.
Thefollowing characteristics may be relevant for microcrystalline
cellulose and carmellose sodium usedas a suspending agent. Transfer 0.250 g to a 125 mL conical flask. Add 25.0 mL of
water Rand 25.0 mL of cupriethylenediamine hydroxide
VIscosity (2.2.10)
solution R. Immediately purge the solution with nitrogen R)
60 per cent to 140 per cent of the nominal value.
insert the stopper and shake until completely dissolved.
Calculate the quantity (x g) needed to prepare exactly 600 g Transfer an appropriate volume of the solution to a suitable
of a dispersion of the stated percentage mlm (dried capillary viscometer (2.2.9). Equilibrate the solution at
substance). To (600 - x) g of water Rat 23-25 °C contained 25 ± 0.1 "C for at least 5 min. Record the flow time (t,) in
in a 1000 mL high-speed blender bowl, add x g of the seconds between the 2 marks on the viscometer. Calculate
substance to be examined and stir at reduced speed, taking
care to avoid contacting the sides of the bowl with the
powder. Continue stirring at low speed for IS s after the
I This monograph has undergone phannacopoeial harmonisation.
See(haprer 5.8 Pharmacopoeial harmonisation.
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2022 Cellulose I-50S
Table 0315.-1. - Intrinsic viscosity table

Intrinsic viscosity [TIleat dIfferent values ofrelative viscosity 'lrd
[oJo
OR' 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
I.I 0.098 0.106 0.115 0.125 0.134 0.143 0.152 0.161 0.170 0.180
1.2 0.189 0.198 0.207 0.216 0.225 0.233 0.242 0.250 0.259 0.268
i.a 0.276 0.285 0.293 0.302 0.310 0.318 0.326 0.334 0.342 0.350
1.4 0.358 0.367 0.375 0.383 0.391 0.399 0.'107 0.414 0.422 0.430
1.5 0.437 0.445 0.453 0.460 0.<168 0.476 0.484 0.491 0.499 0.507
1.6 0.515 0522 0.529 0.536 0.544 0.551 0.558 0.566 0.573 0.580
1.7 0.587 0595 0.602 0.608 0.615 0.622 0.629 0.636 0.642 0.649
I.B 0.656 0.663 0.670 0.677 0.683 0.690 0.697 0.704 0.710 0.717
I.. 0.723 0.730 0.736 0.743 0.149 0.756 0.762 0.769 0.775 0.782
2.0 0.788 0.795 0.802 0.809 0.815 0.821 0.827 0.833 0.840 0.846
2.1 0.852 0.858 0.864 0.870 0.876 0.882 0.888 0.894 0.900 0.906
2.2 0.912 0.918 0.924 0.929 0.935 0.941 0.948 0.953 0.959 0.965
2.' 0.971 0.976 0.983 0.988 0.994 1.000 1.006 1.011 ],017 1.022
2.4 1.028 1.033 1.039 1.044 1.050 1.056 1.061 1.067 1.072 1.078
25 I.OS3 1.089 1.094 1.100 1.105 1.111 1.116 1.121 L.l26 1.131
2.6 1.137 1.142 1.147 1.153 U58 1.I63 1.169 1.174 1.179 1.184
2.7 1.190 1.195 1.200 1.205 1.210 1.215 1.220 1.225 1.230 1.235
2.B 1.240 1.245 1.250 1.255 1.260 1.265 1.270 1.275 1.280 1.285
2.' 1.290 1.295 1.300 1.305 1.310 1.314 1.319 1.324 1.329 1.333
3.0 1.338 1.343 1.348 1.352 1.357 1,362 1.367 l.371 1.376 1.381
3.1 1.386 1.390 1.395 1.400 1.405 1.'109 1.'114 1.418 1,423 1,427
3.2 l.432 1.436 1.441 1.446 1.450 1,455 1.459 1.464 1,468 1.473
3.3 1.477 1.482 1.486 1.491 1.496 1.500 1.504 1.508 1..513 1.517
3.4 1.521 1.525 1.529 1.533 1.537 1.542 1.546 l.550 1.554 1.558
35 1.562 1.566 1.570 1.575 1.579 1.583 1.587 l.591 1.595 1.600
3.6 1.604 1.608 1.612 1.617 1.621 1.625 1.629 1.633 1.637 1.642
'.7 1.646 1.650 1.654 1.658 1.662 1.666 l.671 1.675 1.679 1.683
3.B 1.687 1.691 1.695 1.700 1.704 1.708 1.712 1.715 1.719 1.723
3.' 1.727 1.731 1.735 1.739 1.742 1.746 1.750 1.754 1.758 1.762
4.0 1.765 1.769 1.713 1.777 1.781 1.785 1.789 ],792 1.796 1.800
4.1 1.804 1.808 1.811 1.815 1.819 1.822 1.826 1.830 1.833 1.837
4.2 1.841 1.845 1.848 1.852 1.856 1.859 1.863 1.867 1.870 1.874
4.' 1.878 1.882 1.885 1.889 1.893 1.896 1.900 1.904 1.907 1.911
4.4 1.914 1.918 1.921 1.925 1.929 1.932 1.936 1.939 1.943 1.946
4.5 1.950 1.954 1.957 1.961 1.964 1.968 1.971 1.975 1.979 1.982
4.6 1.986 1.989 1.993 1.996 2.000 2.003 2.007 2.010 2.013 2.017
4.7 2.020 2.023 2.027 2.030 2.033 2.037 2.040 2,043 2.047 2.050
4.8 2,053 2,057 2.060 2.063 2.067 2.070 2.073 2.077 2.080 2.083
4.' 2.087 2.090 2.093 2.097 2.100 2.103 2.101 2.110 2.113 2.116
5.0 2.l19 2.122 2.125 2.129 2.132 2.135 2.139 2.142 2.145 2.148
5.1 2.151 2.154 2.158 2.160 2.164 2.167 2.170 2.173 2.176 2.180
5.2 2.183 2.186 2.190 2.192 2.195 2.197 2.200 2.203 2.206 2.209
5.3 2.212 2.215 2.218 2.221 2.224 2.227 2.230 2.233 2.236 2.240
5.4 2,243 2.246 2.249 2.252 2.255 2.258 2.261 2.264 2.267 2.270
5.5 2.273 2.276 2.279 2.282 2.285 2.288 2.291 2.294 2.297 2.300
5.6 2.303 2.306 2.309 2.312 2.315 2.318 2.320 2.324 2.326 2.329
5.7 2.332 2.335 2.338 2.341 2.344 2.3017 2.350 2.353 2.355 2.358
5.B 2.361 2.364 2.367 2.370 2.373 2.376 2.379 2.382 2.384 2.387
5.' 2.390 2.393 2.396 2.400 2.403 2.0105 2.4OS 2.41 I 2,414 2.417
6.0 2.419 2.422 2.425 2.428 2.431 2.433 2.436 2.439 2.442 2.4014
6.1 2.0147 2.450 2.0153 2.0156 2,458 2.461 2.464 2,467 2.470 2.472
6.2 2.475 2.478 2,481 2,483 2.0186 2.489 2.492 2.494 2,497 2.500
6.3 2.503 2.505 2.508 2.511 2.513 2.516 2.518 2.521 2.524 2.526
6.4 2.529 2.532 2.534 2.537 2.540 2.542 2.545 2.547 2.550 2.553
6.5 2.555 2.558 2.561 2.563 2.566 2.568 2.571 2.574 2.576 2.579
6.6 2.581 2.584 2.587 2.590 2.592 2.595 2.597 2.600 2.603 2.605
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Intrinsic viscosity {'llo at different values of relative viscosity "m
[Il]"
0", 0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09
•.7 2.608 2.610 2.613 2.615 2.618 2.620 2.623 2.625 2.627 2.630
•.8 2.633 2.635 2.637 2.640 2.643 2.645 2.648 2.650 2.653 2.655
•. 9 2.658 2.660 2.663 2,665 2.668 2.670 2.673 2.675 2.678 2.680
7.0 2.683 2.685 2.687 2.690 2.693 2.695 2.698 2.700 2.702 2.705
7.1 2.707 2.710 2.712 2.714 2.717 2.719 2.721 2.724 2.726 2.729
7.2 2.731 2.733 2.736 2.738 2.740 2.743 2.745 2.748 2.750 2.752
7.3 2.755 2.757 2.760 2.762 2.764 2.767 2.769 2.771 2.774 2.776
7.' 2.779 2.781 2.783 2.786 2.788 2.790 2.793 2.795 2.798 2.800
7.5 2.802 2.805 2.807 2.809 2.812 2.814 2.816 2.819 2.821 2.823
7.• 2.826 2.828 2.830 2.833 2.835 2.837 2.840 2.842 2.844 2.847
7.7 2.849 2.851 2.854 2.856 2.858 2.860 2.863 2.865 2.868 2.870
7.8 2.873 2.875 2.877 2.879 2.881 2.884 2.887 2.889 2.891 2.893
7.9 2.895 2.898 2.900 2.902 2.905 2.907 2.909 2.911 2.913 2.915
8.0 2.918 2.920 2.922 2.924 2.926 2.928 2.931 2.933 2.935 2.937
8.1 2.939 2.942 2.944 2.946 2.948 2.950 2.952 2.955 2.957 2.959
8.2 2.961 2.963 2.966 2.968 2.970 2.972 2.974 2.976 2.979 2.981
8.3 2.983 2.985 2.987 2.990 2.992 2.994 2.996 2.998 3.000 3.002
8.' 3.004 3.006 3.008 3.010 3.012 3.015 3.017 3.019 3.021 3.023
8.5 3.025 3.027 3.029 3.031 3.033 3.035 3.037 3.040 3.042 3.044
8,. 3.046 3.048 3.050 3.052 3.054 3.056 3.058 3.060 3.062 3.064
8,7 3.067 3.069 3.071 3.073 3.075 3.077 3.079 3.081 3.083 3.085
8.8 3.OS7 3.089 3.092 3.094 3.096 3.098 3.100 3.102 3.104 3.106
8.9 3.108 3.110 3.112 3.114 3.116 3.118 3.120 3.122 3.124 3.126
9.0 3.128 3.130 3.132 3.134 3.136 3.138 3.140 3.142 3.144 3.146
9.1 3.148 3.150 3.152 3.154 3.156 3.158 3.160 3.162 3.164 3.166
9.2 3.168 3.170 3.172 3.174 3.176 3.178 3.180 3.182 3.184 3.186
9,3 3.188 3.190 3.192 3.194 3.196 3.198 3.200 3.202 3.204 3.206
9.4 3.208 .3.210 3.212 3.214 .3.215 3.217 3.219 3.221 3.223 3.225
9.5 3.227 3.229 3.231 3.233 3.235 3.237 .3.239 3.241 3.242 3.244
9 .e 3.246 3.248 .3.250 3.252 3.254 3.256 3.258 3.260 3.262 3.264
9.7 3.266 3.268 3.269 3.271 3.273 3.275 3.277 3.279 3.281 3.283
9.8 3.285 3.287 3,289 3.291 3.293 3.295 3.297 3.298 3.300 3.302
9.9 3.304 3.305 3.307 3.309 3.311 3.313 3.316 3.318 3.320 3.321
Intrinsic viscosity [ttl ... at different values of relative viscosity 1],...,
[TIle
On> 0.0 0.1 0.2 0.3 0.4 0.5 0.• 0.7 0.8 0.9
10 3.32 3.34 3.36 3.37 3.39 3,41 3.43 3,45 3,46 3,48
II 3.50 3.52 3.53 3.55 3.56 3.58 3.60 3.61 3.63 3.64
12 3.66 3.68 3.69 3.71 3.72 3.74 3.76 3.77 3,79 3.80
13 3.80 3.83 3.85 3.86 3.88 3.89 3.90 3.92 3.93 3,95
I' 3.96 3.97 3.99 ',00 4.02 4.03 '.04 4,06 4.07 4.09
I.
15
'7
4.10
4.23
4.35
'.24
4.36
4.11 4.13
4.25
4.37
4.14
4.27
4.38
4.15
4.28
4.39
4.17
4.29
4.41
4.18
4.30
4.42
4.19
4.31
4.43
4.20
4.33
4.44
4.22
'.34
4,45
18 4.46 4.47 4.018 4,49 4.50 4.52 4.53 4.54 4.55 4.56
19 4.57 4.58 4.59 '.60 4.61 4.62 4,63 '.64 4.65 4.66
the kinematic viscosity (VI) of the solution using the following expression:
expression:
,,(k,)
,,(kd
k, viscometer constant.
k, viscometer constant.
Determine the relative viscosity (l1rd) of the substance to be
Dilute a suitable volume of cupn'ethylenediamine hydroxide examined using the following expression:
solution R with an equal volume of water R and measure the
flow time- (Ii) using a suitable capillary viscometer. Calculate v';v,
the kinematic viscosity (V2) of the solvent using the following
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2022 Cellulose Acetate 1-507
Determine the intrinsic viscosity ([If],) by interpolation,using criteria. In such cases) a cross-reference to the tests described in the
the intrinsic viscosity table (Table 0315.-1). mandatory part is included in the Functionality-related
Calculate the degree of polymerisation (P) using the characteristics section. Control of the characteristics can contribute
following expression: to the qualityof a medicinal product by improving the consistency
of the numufactuting process and theperjonnance of the medicinal
product duringuse. lfIhere control methods are dud, they are
m[(IOO - b)/IOOj recognised as being suitable for the purpose, but othermethods can
m mass of the substance 10 be examined, in grams; reported, the control metlwdmust be indicated.
b loss on drying, in per cent.
Thefollowing characteristics may be relevom for powdered cellulose
usedas diluent or disintegram.
TESTS
Loss on drying
OSolublllty
(see Tests).
Dissolve 50 mg in 10 mL of ammoniacal solution of copper
tetrammine R. It dissolves completely, leaving no residue.O Particle-size distribution (1.9.31 or 1.9.38)
pH (1.1.3) Powder flow (1.9.36)
5.0 to 7.5 for the supernatant. o
_______ PhE"'
Mix 10 g with 90 mL of carbon dioxide-free water R and allow
~~
to stand wil:h occasional stirJ:ing for 1 h.

Ether-soluble substances
Maximum 0.15 per cent (15.0 mg) for the difference
between the mass of the residue and the mass obtained from Cellulose Acetate1
a blank determination,
Place 10.0 g in a chromatography column about 20 mm in
internal diameter and pass 50 mL of peroxide-free ether R Action and use
through the column. Evaporate the eluate to dryness in a Excipient.
previously dried and tared evaporating dish, with the aid of a PhEu _~ _
current of air in a fume cupboard. After all the ether has
evaporated, dry the residue at lOS °C for 30 min, allow to DEFINITION
cool in a desiccator and weigh. Carry out a blank Partly or completely O-acetylated cellulose.
determination. . Content
Water-soluble substances - acetyl groups (C,H,O; M, 43.04): 29.0 per cent to
Maximum 1.5 per cent (15.0 mg) for the differeoce between 44.8 per cent (dried substance), 90.0 per cent to
the mass of the residue and the mass obtained from a blank 110.0 per cent of the nominal content (dried substance).
determination. CHARACTERS
Shake 6.0 g with 90 mL of carbon dioxide-free waterR for Appearance
10 min. Filter with the aid of vacuum into a tared flask. White, yellowish-white or greyish-white, hygroscopic powder
Discard the /irst 10 mL of the filtrate and pass the filtrate or granules.
through the same filter a second time" if necessary, to obtain
Solubility
a clear filtrate. Evaporate a 15.0 mL portion of the filtrate to
Practically insoluble in water, soluble in acetone) in formic
dryness in a tared evaporating dish without charring. Dry at
acid and in a mixture of equal volumes of methanol and
lOS °C for I h, allow to cool in a desiccator and weigh.
methylene chloride, practically insoluble in ethanol
Carry out a blank determination.
(96 per cent).
Maximum 6.5 per cent, determined on 1.000 g by drying in IDENTIFICATION
an oven at 105 °C for 3 h. Infrared absorption spectrophotometry (1.1.14).
Sulfated ash (1.4.14) Comparison cellulose acetate CRS.
Maximum 0.3 per cent (dried substance), determined on Preparation . Prepare a 20 gIL solution of cellulose acetate)
1.0 g. previously dried, in acetone R (mono- or diester) or in
methylene chloride R (di- or triester), and spread I drop of the
tMicroblal contamination
solution between 2 sodium chloride plates; separate the
TM,.\C: acceptance criterion 10' CFU/g (1.6.11).
plates" heat them both at 105 °C for 1 h, and reassemble the
TYMC: acceptance criterion 10' CFU/g (1.6.JZ). dried plates.
TESTS
Absence of Pseudomonas aeruginosa (1.6.11).
Free acid
Absence of Staphylocoa:us aureus (1.6.13). .Maximum 0.1 per cent, calculated as acetic acid (dried
Absence of Salmondla (1.6.11).• substance).
OFUNCTIONALITY-RELATED CHARACTERISTICS To 5.00 g in a 250 mL conical flask, add 150 mL of carbon
This section provides infonnation on characteristics tha:are dioxide-free waterR" insert the stopper, swirl the suspension
recognised as being relevant control parameters for oneor more gently and allow to stand for 3 h. Filter, then wash the flask
junctions of the substance when usedas an excipient (see chapter and the filter with carbon dioxide-free water R) adding the
related characteristics section may also bepresent in the mandatory I This monograph has undergone pharmacopoeial harmonisation.
part of the monograph since they also represent mandatory quality See chapter 5.8 Pharmacopoeial harmonisation:
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1-508 Cellulose Acetate 2022
washings to the filtrate. Add 0.1 mL of phenolphlhalein d loss on drying as a percentage;

m mass of me substance to be examined, in grams;
solution Rl and titrate the combined filtrate and washings
II net amount of 0.5 M sodium hydroxide consumed, in millimoles.
with 0.01 M sodium hydroxide until a pale pink colour is
obtained.
STORAGE
of free acid, calculated as acetic acid.
LABELLING
The label states the nominal percentage content of acetyl
groups.
Sulfated ash (2.4.14) FUNCTIONALITY-RELATED CHARACTERISTICS
Maximum 0.1 per cent, determined on 1.0 g. This seaton provides injormation on characteristics that are
recognised as being relevant control parameters for oneor more
functions of the substance when usedas an excipient (see chapter
TAMC: acceptance criterion 10' CFUlg (2.6.12).
TYMC: acceptance criterion 10 2 CFUlg (2.6.12). related characteristics section may also be present in the mandatory
Absence of Escherichia coli (2.6.13). part of lhe monograph since they also represent mandatory quality
Absence of Sa/moneUa (2.6.13). criteria, In such cases, a cross-reference to the tests described in the
mandatory pan is included in the Functionality-related
ASSAY
characteristics section. ConlTOl of the charactenstia can contrilnue
A. Cellulose acetate containing not more than 42.0 per cent
UJ the qualityof a medicinal product by improving the consistency
of acetyl groups. of the manufacnm'ng process and the perfonnance of the medicinal
To 2.000 g in a 500 mL conical flask, add 100 mL of product duringuse. Where control methods arecited, they are
acetone Rand 5-10 mL of waur R. Close the flask and stir recognised as being suitable for the purpose, but othermethods can
with a magnetic stirrer until dissolution is complete. also be used. Wherever results for a particular characteristic an!'
Add 30.0 mL of 1 M sodium hydroxide with constant stirring. reported, the control method must be indicaud.
A finely divided precipitate of regenerated cellulose, free from
The following characteristics may be relevant for cellulose acetate
lumps, is obtained. Close the flask and stir with a magnetic
usedasfilm former.
stirrer for 30 min. Add 100 mL of waterR at 80°C, washing
down the sides of the flask, stir for 2 min and cool to room Viscosity
temperature. Titrate with 0.5 M sulfuric acid, using 0.1 mL of Dissolve 10 g in a mixture of 50 mL of methanol Rand
phenolphthalein solution R/ as, indicator. Carry out a blank 50 mL of mtlhylene chloride R by shaking. Detennine the
titration. viscosity of this solution at 20 ± 0.1 °C using a rotating
viscometer (2. 2.11J).
Calculate the percentage content of acetyl groups using the
following expression: Acetyl groups
See Assay.
4.305(n, - n,) x 100 Thefollowing characteristics may be relevant for cellulose acetate
(100 - d) x m usedas matrixformer in prokmged-release tableu.
Viscosity
d loss on drying as a percentage;
m mass of the substance to be examined, in grams;
See test above.
111 volume of 0.5 M sulflln, acid used in the titration, in millilitreSj Acetyl groups
112, volume of 0.5 M sulfuric acid used in the blank titration, in See Assay.
millilitres.
Molecular mass distribution (2.2.31J)
B. Cellulose acetate containing more than 42.0 per cent of Particle-size distribution (2.9.31)
acetyl groups.
To 2.000 g in a 500 mL conical flask, add 30 mL of dimethyl _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
sulfoxide R and 100 mL of a«Unle R. Close the flask and srir
with a magnetic stirrer for 16 h. Add 30.0 mL of 1 M sodium
hydroxide with constant stirring. Close the flask and stir with
a magnetic stirrer for 6 min. Allow to stand without stirring Cellulose Acetate Butyrate
for 60 min. Resume stirring and add 100 mL of waterRat
80°C, washing down the sides of the flask, stir for 2 min (ph. Bur. monograph 1406)
and cool to room temperature. Titrate with 0.5 M
Action and use
hydrochloric acid, using 0.1 mL of phenolphlhalein solutwn Rl
Excipient.
as indicator. Add 0.5 mL of 0.5 M hydrochloric acid in excess,
stir for 5 min and allow to stand for 30 min. Titrate with PhE" _
0.5 M sodium hydroxide until a persistent pink colour is
obtained, stirring with a magnetic stirrer. Calculate the net DEFINITION
amount of 0.5 M sodium hydroxide consumed, in mlllimoles, Partly or completely Ccacerylated and O-butyrated cellulose.
taking the mean of 2 blank. titrations into consideration. Content
Calculate the percentage content of acetyl groups using the - a«t:JI groups (C 2H,O): 2.0 per cent to 30.0 per cent
following expression: (dried substance); 90.0 per cent to 110.0 per cent of that
stated on the label (dried substance);
4.305 x n x 100 - butyryl groups (C,H7 0 ): 16.0 per cent to 53.0 per cent
(100 - d) x m (dried substance); 90.0 per cent to 110.0 per cent of mat
stated on the label (dried substance).
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2022 Cetirizine Hydrochloride 1-509
CHARACfERS Injection 20 ~L.

Appearance Calculate the percentage content of acetic acid and butyric
White, yellowish-white or greyish-white powderor granules, acid using the chromatograms obtained with me 2 solutions.
slightly hygroscopic. To calculate the percentage content of acetyl (C 2H30) and
Solubility of butyryl (C 4H70) groups, multiply the percentage content
Practically insoluble in water, soluble in acetone, in lonnie of acetic acid and butyric acid by 0.717 and 0.807,
acid and in a mixture of equal volumes of methanol and respectively.
methylene chloride, practically insoluble in ethanol STORAGE
(96 per cent). In an airtight container.
IDENTIFICATION LABELLING
A. Infrared absorption spectrophotometry (2.2.24). The label states the nominal percentage content of acetyl and
Comparison cellulose autate butyrate CRS. butyryl groups.
The intensity of the bands may vary according [0 the degree _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
of substitution.
B. It complies with the limits of the assay.
TESTS
Acidity Cetirizine Hydrochloride
To 5.00 g in a 250 mL conical flask, add 150 mL of carbon
(Cetirizine Dihydrochloride, Ph. Bur. monograph
dioxide-free water R, insert the stopper, swirl the suspension
1084)
gently and allow to stand for 3 h. Filter, wash the flask and
the fiberwith carbon dioxide-free waterR. Combine the filttate
and washings. Add 0.1 mL of phenolphthalein solution RI. cIDxrN~O",-/Co,H
Not more than 3.0 mL of 0.01 M sodium hydroxide is I ~ N~
required to change the colour of the indicator.
HO .
and enanUomer
,2HCI

Total ash (2.4.16) 461.8 83881-52-1
Maximum 0.1 per cent..
Action and use
ASSAY Histamine HI receptor antagonist; antihistamine.
Liquid chromatography (2.2.29). Preparadons
Test solution To 1.000 g of the substance to be examined in Cetirizine Capsules
a 500 mL conical flask, add 100 mL of acetone R and 10 mL
Cetirizine Oral Solution
of water R. Close the flask and stir with a magnetic stirrer
until dissolution is complete. Add 30.0 mL of 1 M,odium Cetirizine Tablets
hydroxide with constantstirring. Close the flask and stir with PhE" _
a magnetic stirrer for 30 min. Add 100 mL of hot water Rat
80 °C, washing down the sides of the flask and stir for DEFINITION
2 min. Cool, centrifuge or filter the suspension and wash the (RSj-2- [2-[4-[(4-Chlorophenyl)phenylmethyl] piperazin-I-yl)
residue with water R. Combine the filtrate and washings, etboxy]acetic acid dihydrochloride.
adjust to pH 3 with dilute phosphoric acidR and dilute to Content
500.0 mL with waterR. 99.0 per cent to 101.0 per cent (dried substance).
Reference solution Dissolve 0.200 g of gladal acetic acid R CHARACfERS
and 0.400 g of butyric acid R in water RJ adjust to pH 3 with Appearance
dilute phosphoric acidR and dilute to 500.0 mL with waterR. White or almost white powder.
Column: Solubility
- size: 1 = 0.25 m, 0 :::: 4.6 mm; Freelysoluble in water, practically insoluble in acetone and
- stationary phase: octadecylsilyl ,ui<a gelfor chromatography R in methylene chloride.
(5 urn).
Mobile phase: IDENTIFICATION
- mobile phase A: methanol R; Firs' identificaticn: B, D.
- mobue phaseB: phosphate buffer sdudon pH 3.0 R1; Second identifica'imr: A, C, D.
Tim. Mobile phase A MobUe phase B (2.2.25).
Testsolution Dissolve 20.0 mg in 50 mL of a 10.3 gIL
0-30 5 95 solution of hydrochloric acidR and dilute to 100.0 mf.wirh
30·35 5 --> 20 95 ..... 80
the same acid. Dilute 10.0 mL of this solution to 100.0 mL
35 - 60 20 80 with a 10.3 gIL solution of hydrochloric acid R.
60 - 61 5 95
Spectral range 210-350 om.
Absorption maximum At 231 run.
FIuw rate 1.2 mllmin.
Specific absorbance at the absorption maximum 359 to 381.
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1-510 Cetirizine Hydrochloride 2022
Comparison cetirizine dihydrochloride GRS. Identification of impurities Use the chromatogram supplied
C. Thin-layer chromatography (2.2.27). with cetirizine for peak idenujication CRS and the
Test solution Dissolve 10 mg of the substance [0 be chromatogram obtained with reference solution (c) to identify
examined in water R and dilute to 5 mL with the same the peaks due to impurities B, C, D, E and F; use the
solvent.
identify the peak due to impurity A.
Reference solution (aj Dissolve 10 mg of cetinzine
dihydrochloride CRS in waterR and dilute to 5 mL with the Re/aliveretention With reference to cetirizine (retention
same solvent.
=
time about 9 min): impurity D = about 0.6;
Reference solution (b) Dissolve 10 mg of chlorpheuamine
impurity B =about 0.8; impurity C = about 0.9;
maleate CRS in water R and dilute to 5 mL with the same
impurity E =about 1.2; impurity F =about 1.37;
solvent. Mix 1 mL of the solution and 1 mL of reference
impurity A =about 1.42.
solution (a). System suitability Reference solution (c):
- peak-to-valley ratio: minimum 5, where Hp = height above
Plare TLC silica gel OFzs4 plare R. the baseline of the peak due to impurity C and
Mobile phase ammonia R, methanol R J methylene chloride R H1,! =
height above the baseline of the lowest point of the
(1:10:90 VIV/V). curve separating this peak from the peak due to cetirizine.
Application 5 ~L. Limits:
Development Over 2/3 of the plate. - correction factors: for the calculation of content, multiply
Drying In a current of cold air. the peak areas of the following impurities hy the
Daeaion Examine in ultraviolet light at 254 run.
= =
impurity C 1.9; impurity D 0.6; impurity E 1.3; =
impurity F =1.9;
- impurities A, B~ C, D, E, F: for each impurity, not more
Results The principal spot in the chromatogram obtained than 1.5 times the area of the principal peak in the
with the test solution is similar in position and size to the chromatogram obtained with reference solution (b)
principal spot in the chromatogram obtained with reference (0.15 per cent);
solution (a). - unspedfied impurities: for each impurity, not more than the
D. It gives reaction (a) of chlorides (2.3.1). area of the principal peak in the chromatogram obtained
TESTS
Solution S
(0.3 per cent);
Appearance of solution the chromatogram obtained with reference solution (b)
Solution S is clear (2.2.1) and not more intensely coloured (0.05 per cent).
than reference solution BY, (2.2.2, Method If).
pH (2.2.3) Maximum 0.5 per cent, determined on 1.000 g hy drying in
1.2 to 1.8 for solution S. an oven at 105°C.
liquid chromatography (2.2.29). .Maximum 0.2 per cent, determined on 1.0 g.
Testsolution Dissolve 20 mg of the substance to be ASSAY
the mobile phase.
waterR and 70 volumes of aatone R. Titrate with 0.1 M
Reference solution (a) Dissolve 2 mg of cetitizine sodium hydroxide to the 2n d point of inflexion. Determine the
dihydrochlonik CRS and 2 mg of cetirizine impurity A CRS in end-point potentiometrically (2.2.20). Carry out a blank
the mobile phase and dilute to 50.0 mL with the mobile titration.
I mL of O. I M sodium hydroxide is equivalent to 15.39 mg of
mobile phase.
C21H27C13N203'
100.0 mL with the mobile phase. Dilute 1.0 mL of this STORAGE
solution to 10.0 mL with the mobile phase. Protected from light.
Reference solution (c) Dissolve the contents of a vial of IMPURITIES
cetirizine for peak identification CRS (containing impurities B, Specified impurities A, BJ CJ D, E, F.
C, D, E and F) in 5.0 mL of the mobile phase. Otherdetectable impurities (thefollowing substances would, if
Column: present at a sufficient level, be detected by one orother of the tests
=
- size: / 0.25 m, 0 = 4.6. mm; in the monograph. They are limited by thegeneral acceptance
- stationary phase: silica gelfor chromatography R (5 pm). criterion for otherlunspecified impuniies andlor by thegeueraI
Mobile phase dilute sulfuric add R, waterR, aatonitriJe R monograph Substonces for phanuaceutical use (2034). It is
(0.4:6.6:93 VIV/V). therefore not necessary to identify these impun'ties for
Flow rare I mIJmin. demonstration of compliance. See also 5.10. Control of impurities
in substances for pharmaceutical use) G.
Injection 20 ~L.
Run time 3 times the retention time of cetirizine.
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2022 Cetostearyl Alcohol 1-511
CIDy/'
I
NJ 4'
I NH Cetostearyl Alcohol
HO and enanUomer
Action and use
Excipient.
A. (RS)-I-[ (4-chlorophenyl)phenylmethyl]piperazine, PhE" ~ _
DEFINn10N
Mixture of solid aliphatic alcohols, mainly octadecan-f-ol
(stearyl alcohol; C reH,.O; M, 270.5) and hexadecan-f-ol
(cetyl alcohol; C,oR,.O; M, 242.4), of animal or vegetable
origin.
andenanUOmer
Content
B. (RS)-2- [4-[(4-chlorophenyl)phenylmethyl]piperazin-l-yl) - suarylalaJhol: minimum 40.0 percent,
acetic acid, - sum of the contents of'tearyl akoholand cetyl alcohol:
minimum 90.0 per cent.
CHARACTERS
Appearance
White or pale yellow, wax-like mass,plates,flakes or
granules.
aoo enantiomer Solubility
Practically insoluble in water, soluble in ethanol (96 per cent)
C. (RS)-2-[2-[4-[(2-chlorophenyl)phenylmethyl]piperazin-l- and in light petroleum. When melted, it is miscible with fatty
yljethoxyjacetic acid, oils, with liquid paraffin and with melted wool fat.
IDENTIFICATION
Examine the chromatograms obtained in the assay.
Results The 2 principal peaks in the chromatogram obtained
principal peaks in the chromatogram obtained with the
reference solution.
TESTS
Appearance of soludon
The solution is dear (2.2.1) and not more intensely coloured
than reference solution B. (2.2.2, Method II). To 0.50 g, add
D.l,4-bis[(4-chlorophenyl)phenylmethyl)piperazine, 20 mL of cold ethanol (96 per <en!! R. Boil until dissolution is
complete, then allow to cool.
Melting point (2.2.14)
49 ·C to 56 ·C.
Acid value (2.5.1)
Maximum 1.0.
and enereomer
E. (RS)-2-[2-[2-[4-[(4-chlorophenyJ)phenylmethyl]piperazin- 208 to 228.
I-yl]ethoxy)ethoxy)acetic acid (ethoxycetirizine), Iodine value (2.5.4, MethodA)
Maximum 2.0.
~,""
r N ~O '-./ CO'H Dissolve 2.00 g in methylene chloride R and dilute to 25 mL
I#, NJ with me same solvent.
Saponification value (2.5.6)
Maximum 2.0.
4' ASSAY
Gas chromatography (2.2.28): use the nonnalisation
F. 2-[2-[4-(diphenylmethyl)piperazin-I-yl]ethoxy)acetic acid, procedure.
examined in ethanol (96 per <en!! R and dilute to 10.0 mL
Reference solution Dissolve 60 mg of cetyl alcohol GRS and
40 mg of stearyl alcohol GRS in ethanol (96 per cen!! R and
and enanllomer dilute to J0 mL with the same solvent. Dilute 1 mL of this
solution to 10 mL with ethanol (96 per <en!! R.
G. (RS)-2- [4-[(4-chlorophenyl)phenyhnethyl]piperazin-l-yl) Column:
ethan-l-ol. - size: I;;;; 30 m, 0 ;;;; 0.32 mm,
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" - stationary phase: methylpoly,iloxane R (I urn).
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1-512 Cetostearyl Alcohol 2022
Cartier gas helium for chromatography R. Reference solution (b) Dissolve 20 mg of sodium cetostearyl
Flow rate 1 mUmin. sulfate R in 10 mL of ethanol (70 per cent VII-? R, heating on
Split ratio 1:100. a water-bath.
Temperature: Plate TLC ocladecylsilyl silica gel Pm plateR.
Mobik phase waterR, acewne R, methanol R
Tim, Temperature (20:40:40 VlVfII).
(mIn) ('C) Applicotion 10 ~L.
Column 0-20 150 --> 250 Development Over 2/3 of the plate.
20 - 40 250
Drying In air.
Injectionport 250
Detector 250
Detection Spray with a 50 WL solution of phosphomolybdic
acid R in ethanol (96 per cent) R; heat at 120°C untilspots
appear (about 5 min).
Results:
Injection I ~L. - the 2 principal spots in the chromatogram obtained
System suilabihiy Reference solution: with test solution (a) are similar in positionand colour
- resolution: minimum 5.0 between the peaks due to cetyl to the principal spots in the chromatogram obtained
alcohol and stearyl alcohol. with reference solution (a);
Calculate lite percentage contents of C 1Ji"349 and C JSH3SO. - 2 of the spots in the chromatogram obtained with rest
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" solution (b) are similar in positionand colourto the
principal spots in the chromatogram obtained with
B. Examine the chromatograms obtained in the assay of
Emulsifying Cetostearyl Alcohol cetostearyl alcohol.
(Type A) with the test solution are similar in retention time to the
(Ph. Hur. monograph 0801) 2 principal peaks in the chromatograms obtained with
reference solutions (a) and (b).
Action and use
Excipient. C. It gives a yellow colour to a non-luminous flame.
D. To 0.3 g add 20 mL of anhydrous ethanol Rand heat to
PhE" -'- _
boilingon a water-bath with shaking. Filter the mixture
DEFINITION immediately, evaporate to dryness and take up the residue in
Mixture of cetostearyl alcohol and sodium cetostearyl sulfate. 7 mL of water R. To I mL of the solution add 0.1 mL of a
A suitable buffer may be added. I WL solution of methylene blue R, 2 mL of dilute su/furi£
acid Rand 2 mL of methylene chloride R and shake. A blue
Content
colour develops in the lowerlayer.
- cetostearyl alcohol: minimum 80.0 per cent (anhydrous
substance); TESTS
- sodium cetosteary/ sulfate: minimum 7.0 per cent Acid value (2.5.1)
(anhydrous substance). Maximum 2.0.
CHARACTERS Iodine value (2.5.4, MethodA)
Appearance Maximum 3.0.
White or pale yellow, waxy mass, plates, flakes or granules. Dissolve 2.00 g in 25 mL of methylene chloride R.
Solubility Saponification value (2.5.6)
Soluble in hot water giving an opalescent solution, practically Maximum 2.0.
insoluble in cold water, slightly soluble in ethanol Water (2.5.12)
(96 per cent). Maximum 3.0 per cent, determined on 2.50 g.
IDENTIFICATION ASSAY
Firsr identification: B, C, D. Cetoslearyl alcohol
Second identification: A, C, D. Gas chromatography (2.2.28).
A. Thin-layer chromatography (2.2.27). Internal standard solution Dissolve 0.200 g of
Test solution (a) Dissolve 0.1 g of the substance to be l-nonadecanol CRS in anhydrous ethanol R and dilute to
examined in 10 mL of trimethylpetltane R, heating on a water- 100.0 mL with the same solvent.
bath. Shake with 2 mL of ethanol (70 per unt VII-? Rand Test solution Dissolve 0.200 g of the substance to be
allow to separate. Use the lower layer as test solution (b). examined in 25.0 mL of the internal standard solution.
Dilute 1 mL of the upperlayerto 8 mL with Add 25 mL of water R and shake with 4 quantities, each of
trimethylpentane R. 25 mI.., of pentane R, adding sodium chloride R, if necessary,
Test solution (b) Use the lower layer obtained in the me
to facilitate separation of the layers. Combine the upper
preparation of test solution (a). layers, wash with 2 quantities) each of 30 ml., of water R, dry
Reference solution (a) Dissolve 24 mg of ceryl alcohol CRS over anhydrous sodium sulfate R and filter.
and 16 mg of stearyl alcohol CRS in 10 mL of Reference solution (a) Dissolve 0.100 g of ceryl alwholCRS
trimethylpentane R. in 25.0 mL of the internal standard solution. Add 25 mL of
water R and shakewith 4 quantities, each of 25 mL, of
pentane R, adding sodium chloride R, if necessary, to facilitate
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2022 Cetostearyl Alcohol 1-513
the separation of the layers. Combine the upper layers, wash The percentage content of cetostearyl alcohol corresponds to
with 2 quantities, each of 30 mL, of water R, dry over the sum of the percentage contents of cetyl alcohol and
anhydrous sodium sulfate R and filter. stearyl alcohol.
Reference solution (b) Dissolve 0.1 00 g of Sleary! alcohol CRS Sodium cetostearyl sulfate
in 25.0 mL of the internal standard solution. Add 25 mL of Disperse 0.300 g in 25 mL of methylene chloride R.
waterR and shake with 4 quantities, each of 25 ml., of Add 50 mL of water R and 10 mL of dimidium bromide-ndfan
pentane R, adding sodium chloride R, if necessary, to facilitate blue mixed solution R. Titrate with 0.004 lH benzethonium
the separation of the layers. Combine the upper layers, wash chloride, using sonication, heating, and allowing the layers to
with 2 quantities, each of 30 mI..., of water R, dry over separate before each addition, until the colour of the lower
anhydrous sodium sulfate R and filter. layer changes from pink. to grey.
Column: 1 mL of 0.004 M benzethonium chloride is equivalent to
- material: fused silica; 1.434 mg of sodium cetoatearyl sulfate,
- size: 1= 25 ill, 0 = 0.25 mm;
LABELLING
- stationary phase: methylpo/y,iloxa"e R (film thickness
The label states, where applicable, the name and
0.25 um).
concentration of any added buffer.
Comer gas helium for chromatography R. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Fluw rate I mllmin.
Sphi ratio 1:100.
Temperaiu...:
Emulsifying Cetostearyl Alcohol
Tim. Temperature
(mln) eel (Type B)
Column 0·20 150 --J 250 (ph. Bur. monograph 0802)
Injection port 2"
Detector 2'0
Action and use
Excipient.
Detection Flame ionisation. PhE" _
Injeaion I ~L.
DEFINITION
Elution order Cetyl alcohol, stearyl alcohol, I-nonadecanol. Mixture of cetcstearyl alcohol and sodium laurilsulfate.
Calculate the percentage' content of cetyl alcohol in the A suitable buffer may be added.
substance to be examined using the following expression and Content
taking into account the assigned content of cetylalaJhol CRS: - ceto'teary! alcohol: minimum 80.0 per cent (anhydrous
substance);
A xA- xmr,y
- x -1x l 00
2
- sodium laurilsulfate: minimum 7.0 per cent (anhydrous
x AI Ax,y m substance).
A,,- area of the peak due to cetyl alcohol in the chromatogram CHARACTERS
obtained with the test solution; Appearance
A,,-..,. area of the peak due to cayI '*"1101 CRS in the chromatogram White or pale yellow, waxy mass, plates, flakes or granules.
obtained with reference solution (a);
AI area of the peak due to the internal standard in the Solubility
chromatogram obtained with the test solution; Soluble in hot water giving an opalescent solution, practically
A2 area of the peak due to the internal standard in the
insoluble in cold water, slightly soluble in ethanol
chromatogram obtained with reference solution (a);
HI mass of the substance to be examined in the test solution, in (96 per cent).
milligrams;
IDENTIFICATION
m,,-,y mass of cerylakohbl CRS in reference solution (a), in milligrams.
First identification: B, C, D.
Calculate the percentage content of stearyl alcohol in the Second identification: A, C, D.
substance to be examined using the following expression and A. Thin-layer chromatography (2.2.27).
taking into account the assigned content of stearyl Test solution (aJ Dissolve 0.1 g of the substance to be
a1<:ohoiCRS: examined in 10 mL of trimelhylpenume R, heating on a water-
bath. Shake with 2 mL of etha,,01 (70 per cent VII-? Rand
A, m I
A~x - x~ x - x 100 allow to separate. Use the lower layer as test solution (b).
AI Az,y m Dilute I mL or the upper layer to 8 mL with
tllmethylpentane R.
A... area of the peak due to stearyl alcohol in the chromatogram
obtained with the test solution; Test solrttion (b) Use the lower layer obtained in the
Az,y area of the peak due to SIaJ~ alcohol CRS in the chromatogram preparation of test solution (a),
Al area of me peak due to the internal standard in the
Reference solution (a) Dissolve 24 mg of cetyl alcohdCRS
chromatogram obtained with the test solution; and 16 mg of maryl a1<:ohol CRS in 10 mL of
AJ area of the peak due to lite internal standard in the trimethy/penta"e R.
chromatogram obtained with reference solution (b)j
m mass of the substance 10 be examined in the test solution, in
Reference solutio" (b) Dissolve 20 mg of sodium
mlljigrams; lauri/sulfate CRS in 10 mL of ethanol (70 per ce"t VII-? R,
nJ",y mass of stealji alcoholCRS in reference solution (b), in heating on a water-bath.
milligrams. Plate TLC octade<:y/sllyl silica gelF1S4 plate R.
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1-514 Cetostearyl Alcohol 2022
klobiJe phase water R, tuetone R, methanol R Reference solution (b) Dissolve 0.100 g of maryl akohol GRS
(20:40:40 VIVIV). in 25.0 mL of the internal standard solution. Add 25 rnL of
Applkaticn IO ~L. water R and shake with 4 quantities, each of 25 mL, of
Development Over 213 of the plate. pentane R, adding sodium chloride R, if necessary) to facilitate
the separation of the layers. Combine me upper layers, wash
Drying In air. with 2 quantities, each of 30 ml., of water R, dry over
Detection Spray with a 50 gIL solution of phosphomolybdic anhydrous sodium sulfateR and filter.
acid R in ethanol (96 percent.! R; heat at 120 'C until spots Golumn:
appear (about 5 min). - material: fused silica;
Results: - size: 1= 25 m, 0 =
0.25 mm;
- the 2 principal spots in the chromatogram obtained - stationary phase: methy/po/ysiloxane R (film thickness
with test solution (a) are similar in position and colour 0.25 urn).
to the principal spots in the chromatogram obtained Carrier gas heliumfor chromatography R.
with reference solution (a),
- 1 of the spots in the chromatogram obtained with test Flow rate I mIlrnin.
solution (b) is similar in position and colour to the Sp/,i rauo 1:100.
principal spot in the chromatogram obtained with Temperature:
B. Examine the chromatograms obtained in the assay of Time Temperature
cetosrearyl alcohol,
(min) CCJ
Column 0-20 150 ..... 250
Injection port 250
with the test solution are similar In retention time to the
Detector 250
2 principal peaks in the chromatograms obtained with
reference solutions (a) and (b).
C. It gives a yellow colour to a non-luminous flame.
D. To 0.3 g add 20 rnL of anhydrous ethanol R and heat to
Injection I J.lL.
boiling on a water-bath with shaking. Filter the mixture E/uu"on order Cetyl alcohol, stearyl alcohol, J-nonadecanol.
immediately, evaporate t.o dryness and take up the residue in Calculate the percentage content of cetyl alcohol in the
7 mL of waterR. To 1 rnL of the solution add 0.1 mL of a substance to be examined using the following expression and
1 gIL solution of methylene blueR, 2 mL of dilute sulfuric taking into account the assigned content of cetyl akohol CRS:
acid R and 2 mL of methylerie chloride R and shake. A blue
colour develops in the lower layer.
TESTS
Acid value (2.5.1) A", area of the peak due to cetyl alcohol in the chromatogram
Maxiroum 2.0. obtained wirh the test solution;
A",,,. area of the peak due to cetylalcoholCRS in the chromatogram
Iodine value (2.5.4, Method A)
obtained wirh reference solution (a);
Maximum 3.0. AI area of the peak due to the internal standard in the
Dissolve 2.00 g in 25 mL of methylene chloride R. chromatogram obtained wirh the test solution;
A2 area of the peak due to me internal standard in the
Saponification value (2.5.6) chromatogram obtained with reference solution (a);
Maximum 2.0. m mass of the substance to be examined in the test solution. in
milligrams;
Water (2.5.12) m~". mass of ~ dko!wlCRS in reference solution (a). in milligrams.
ASSAY Calculate the percentage content of stearyl alcohol in the
Cetostearyl alcohol substance to be examined using the following expression and
Gas chromatography (2.2.28). taking into account the assigned content of stearyl
akoholGRS:
Internalstandardsolution Dissolve 0.200 g of
1-nonadecanol GRS in anhydrous ethanol R and dilute to
A n1 ,y J
100.0 mL with the same solvent. A x - 3 x - z x - x 100
Z Al All'eY m
Test sdution Dissolve 0.200 g of the substance to be
examined in 25.0 mL of the internal standard solution. A~ area of the peak due to stearyl alcohol in the chromatogram
Add 25 mL of water R and shake with 4 quantities, each of obtained with the test solution;
25 mL, of pentane R, adding sodium chloride R, if necessary, A.". area oCthe peak due to Slearyl a1uJhol CRS in the chromatogram
obtained with reference solution (b)j
t.o facilitate the separation of the layers. Combine me upper
AI area. of the peak due to the internal standard in the
layers, wash with 2 quantities, each of 30 mL, of walei' R, dry chromatogram obtained with the [est solution;
over anhydrous sodium sulfate R and filter. A) area of the peak due to the internal standard in the
chromatogram obtained with reference solution (b);
Reference soluticn (a) Dissolve 0.100 g of cetylakohol GRS
m mass of me substance to be examined in me test solution, in
in 25.0 mL of the internal standard solution. Add 25 mL of milligrams;
water R and shake with 4 quantities, each of 25 mL, of m.". mass of sreary!dkchoi CRS in reference solution (b), in
pentane R, adding sodium chloride R, if necessary, to facilitate milligrams.
the separation of the layers. Combine the upper layers, wash
with 2 quantities, each of 30 ml., of water R, dry over The percentage content of cetosrearyl alcohol corresponds to
anhydrous sodium sulfate R and filter. the sum of the percentage contents of cetyl alcohol and
stearyi alcohol.
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2022 Cetrimide 1-515
Sodium laurilsulfate Water (2.5.12)

Disperse 0.300 g in 25 mL of methylene chloride R. Maximum 0.2 per cent, determined on 10.0 g.
Add 50 mL of water Rand 10 mL of dimidium bromide-sulfan Total ash (2.4.16)
blue mixedsoluu'on R. Titrate with 0.004 M beneethonium Maximum 0.2 per cent, determined on 2.0 g.
chloride, using sonication, heating, and allowing the layers to _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
separate beforeeach addition, untillhe colour of the lower
layer changes from pink to grey.
1 mL of 0.004 M benzethonium chloride is equivalent to
1.154 mg of sodium laurilsulfate. Cetrimide
LABELLING
(ph. E<rr. monograph 0378)
The label states, where applicable, the name and
concentration of any added buffer.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
Action and use

Antiseptic.
Cetostearyl Isononanoate Preparations
Cetrimide Cream
Cetrimide Emulsifying Ointment
Action and use
Strong Cettimide Solution
Excipient.
PhE<I _
PhE<I ~ _
DEFINITlON
DEFINITION
Cetrimide consists of trimethyltetradecylammoniurn bromide
Mixture of esters of cetostearyJ alcohol with isononanoic acid,
and may contain smaller amounts of dodecyl- and hexadecyl-
mainly 3,S,5-trimethylhexanoic acid.
trimethylammonium bromides.
CHARACTERS
Content
Appearance 96.0 per cent to 101.0 per cent ofalkyltrimethylammonium
Clear, colourless or sligh!ly yellowish liquid. bromides, calculated as C"H3sBrN (M, 336.4) (dried
Solubility substance).
Practically insoluble in water, soluble in ethanol (96 per cent)
CHARACTERS
and in light petroleum, miscible with fatty oils and with
Appearance
liquid paraffins.
White or almost white) voluminous, free-flowing powder.
Viscosity
Solubility
15 mf'a-s to 30 ml'a.s.
Freely soluble in water and in alcohol.
Relative density
0.85 to 0.86. IDENTIFICATION
A. Dissolve 0.25 g in alcohol R and dilute to 25.0 mL with
Refractive index
the same solvent. At wavelengths from 260 om to 280 nm,
1.44 to 1.45.
the absorbance (2.2.25) of the solutionhas a maximum
IDENTIFICATION of 0.05.
A. On cooling, turbidity occurs below 15 °C. B. Dissolve about 5 mg in 5 mL of buffer solution pH 8.0 R.
B. Saponification value (see Tests). Add about 10 mg of potassium ferricyanide R. A yellow
C. Infrared absorption spectrophotometry (2.2.24). precipitate is formed. Prepare a blank in the same manner
Comparison Ph. Bur. reference spectrum of cetostearyl but omitting the substance to be examined: a yellow solution
is observed but no precipitate is formed,
isononanoate.
C. Solution S (see Tests) froths copiously when shaken.
TESTS
Appearance
The substance to be examined is clear (2.2.1) and not more Test solution Dissolve 0.10 g of the substance to be
intenselycoloured than reference solution Y 6 examinedin water R and dilute to 5 mL with the same
(2.2.2, Method l). solvent.
Acid value (2.5.1) Reference solution Dissolve 0.10 g of
Maximum 1.0, determined on 5.0 g. tn'methyltetradecylammonium bromide CRS in waterR and
Hydroxyl value (2.5.3, Method A)
Plate TLC silanised silica gel Pm plate R.
Maximum 5.0, determined on 4.0 g. Add 5.0 mL of
acetylating reagent. Alobile phase acetone R, 270 gIL solution of sodium acetate R,
methanol R (20:35:45 VIVIJI).
Iodine value (2.5.4, Method A)
Maximum 1.0. Applica'ion I~.
Development Over a pam of 12 cm.
135 to 148, determined on 1.0 g. Drying In a current of hot air.
Detection Allow to cool; expose the plate to iodine vapour
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1-516 Strong Cetrimide Solution 2022
Result The principal spot in the chromatogram obtained PRODUCTION

with the test solution is similar in position, colourand size to In making Strong Cetrimide Solution, Ethanol (96 per cent)
the principal spot in the chromatogram obtained with the maybe replaced by Industrial Methylated Spirit, provided
reference solution. that the law and the statutory regulations governing the use
E. It gives reaction (a) of bromides (2.3. I). of Iodustrial Methylated Spirit are observed.
TESTS Content of cetrimide C17H3SBrN
Solution S 95.0 to 105.0% of the stated amount.
Dissolve 2.0 g in carbon dioxide-free water R and dilute to IDENTIFICATION
100 mL with the same solvent. A. Dilute a volume of the solutioncontaining 0.1 g of
Appearance of solution cetrimide to 5 rnL with water and add 2 rnL of a 5% wlv
Solution S is clear (2.2.1) and colourless (2.2.2, Method11). solutionof potassium hex00'anofen'a~(lll). A yellow precipitate
Acidity or a1kaIInlty is produced.
To 50 mL of solution S add 0.1 mL of bromocresol purple B. Shake together 5 mL of water, 1 mL of 2M sulfuric acid,
solution R. Not more than 0.1 mL of 0.1 M hydrochloric acid 2 mL of chlorofonn and 0.05 mL of methylorange solution; the
or 0.1 .l\-I sodium hydroxide is required to change the colourof chloroform layer is colourless. Add 0.1 mL of the solution
the indicator. being examined and shake; a yellowcolour is produced
Amines and amine salts slowly in the chloroform layer.
Dissolve 5.0 g in 30 mL of a mixture of 1 volwne of 1 M C. Yields reaction A characteristic of bromides, Appendix VI.
hydrochloric acid and 99 volumes of methanol R and add TESTS
100 mL of 2-propanol R. Pass a stream of nitrogen R slowly
Acidity or a1kaIInlty
through the solution. Gradually add 15.0 mL of 0.1 M
Dilute a volume of the solutioncontaining 109 of cetrimide
tetrabutylammonium hydroxide and record the
potentiometric titration curve (2.2.20). If the curve shows
'0 100 mL and add 0.1 mL of bromocresol purple solution.
NOl more than 1.0 mL of either O. 1M hydrochloric acid VS or
2 points of inflexion, the volume of titrant addedbetween the
0.1 M sodium hydroxide VS is required to change the colourof
2 points is not greater than 2.0 rnl.,
the solution.
Loss on dryIng (2.2.32)
MIscibility with ethanol
Mix a volume of the solution containing 1.6 g of cetrimide
with a mixture of 2 mL of water and 16 mL of ethanol
Sulfated ash (2.4.14) (96%). The solution remains clear, Appendix IV A.
Neutral substances
ASSAY To a volume of the solution containing 10 g ofcetrimide add
Dissolve 2.000 g in waU' R and dilute to 100.0 mL with the 25 mL of ethanol (50%), acidify to bromophenol blue solution
same solvent. Transfer 25.0 mL of the solution to a by the drop wise addition of hydrochloric acid and add
separating funnel, add 25 mL of chloroform R, 10 mL of 0.05 mL in excess. Transfer quantitatively £0 the extraction
O. I M sodium hydroxide and 10.0 mL of a freshly prepared compartment of an apparatus designed for continuous liquid-
50 gIL solutionof potassium iodide R. Shake,allow to separate liquid extraction by fluids of a lesserdensity than water,
and discard the chloroform layer. Shake the aqueous layer washing out the beaker with 10 mL ethanol (50%) and
with 3 quantities, each of 10 mL, of chlorofonn R and discard adding the washings 10 the bulk of the solution in the
the chloroform layers. Add 40 mL of hydrochlaric ocidR, extractor. Add sufficient ethanol (50%), if necessary, to half-
allow to cool and titrate with 0.05 M potassium iodate until fill the extraction chamber to the level of the overflow limb.
the deep browncolour is almostdischarged. Add 2 mL of Add sufficient purified hexane to fill the extraction chamber,
chloroform R and continue the titration, shaking vigorously, secure an overflow volumeof about 30 rnL in the ebullition
until the colourof the chloroform layer no longer changes. flask and heat using an electrically heated mantle. Ensure
Carry out a blank titration on a mixture of 10.0 mL of the that a continuous flow of hexane through the aqueous
freshly prepared 50 gIL solution of potassium iodide R, 20 mL ethanol layer is observed and continuethe extraction for
of warer Rand 40 mL of hydrochlori< acidR. 16 hours. Transfer the hexane extract to a separating funnel,
I mL of 0.05 M potassium iodau is equivalent to 33.64 mg of washing out the flask with 10 mL of purified hexane. Shake
C I1H3SBrN . the combined extract and washings with 25 mL of ethanol
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pf>E<I (50%) and discard the aqueous ethanol layer. Filter the
hexane layer through a dry filter paper (Whatman No.1 is
suitable) into a tared flask and removethe solvent using a
rotary evaporator at 40° and then at room temperature at a
Strong Cetrimide Solution pressure not exceeding 0.7 kPa for 2 hours. The residue
weighs not more than0.4 g.
Action and use
Antiseptic. Non-quaternised amines
To a volumeof the solution containing 10 g of cetrimide add
Preparation a mixture of 100 mL of propan-2-ol, 0.1 mL of hydrochloric
Cetrimide Solution
add and 20 mL of methanol. Titrate with
DEFINITION O.lM lelTalmtylammonium hydroxide VS passing a slow current
Strong Cetrimide Solution is an aqueous solutionof of nitrogen through the solutionand determining the end
cetrimide. It contains 20 to 40% wlv of cetrimide, calculated point potentiometrically using a platinum-glass electrode
as CI7H38BrN. It contains Ethanol (96 per cent) or system. Inflections in the titration curve indicate (A)
Isopropyl Alcohol or both. It may be perfumed and may neutralisation of excess hydrochloric acid and (B)
containcolouring matter. neutralisation of non-quatemised amine salts. The difference
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2022 Cetyl Alcohol 1-517
between the volumes corresponding to A and B is not more Solubility

than 10 mL (2.4%, calculated as C"#,,N). Practically insoluble in water, freely soluble or sparingly
Ethanol; Isopropyl alcohol soluble in ethanol (96 per cent). W'hen melted, it is miscible
Carry out one or both of the following methods according to with vegetable and animal oils, with liquid paraffin and with
the declared alcohol content of the solution being examined. melted wool fat.
Ethanol Not more than 10.0% vlv, by the method for the IDENTIFICATION
determination 0/ethanol, Appendix VITI F. Use on-column Examine the chromatograms obtained in the assay.
injection and do not heat the injection pan. Results The principal peak in the chromatogram obtained
Isopropyl alcohol Not more than 10.0% vlv, by the method with the test solution is similar in retention time to the
for the determination of ethanol, Appendix VIII F, with the principal peak in the chromatogram obtained with reference
following modifications. For solution (1) use a solution solution (a).
containing 5.0% vtv of propan-2-01 and 5.0% vlv of propan-l-
TESTS
01 (internal standard). For solution (2) use the solution being
examined, diluted with water, if necessary, to contain about
5.0% v/v of isopropylalcohol. Maintain the column
than reference solution B6 (2.2.2, Method II).
temperature at 170°, use on-column injection and do not
heat the injection port. Dissolve 0.50 g in 20 mL of boiling ethanol (96 per cen!! R.
Allow to cool.
ASSAY
Melting point (2.2.14)
Dilute a volume containing 4 g of cetrimide with sufficient
46°C to 52 °C.
water to produce 100 mL. Transfer 25 mL of the solution to
a separating funnel and add 25 mL of chloroform, 10 mL of Acid value (2.5.1)
O.IM sodium hydroxide and 10 mL ofa freshly prepared Maximum 1.0.
8.0% w/v solution of potassium iodide. Shake well, allow to Hydroxyl value (2.5.3, MethodA)
separate and discard the chloroform layer. Wash the aqueous 218 to 238.
layer with-three 10 mL quantities of chloroform and discard Iodine value (2.5.4, MethodA)
the washings. Add 40 mL of hydrochloric add, cool and titrate Maximum 2.0.
with O. 05M potassium iodate VS until the deep brown colour is
Dissolve 2.00 g in methylene chloride R and dilute to 25 mL
almost discharged. Add 2 mL of chlorofonn and continue the
titration, with shaking, until the chloroform layer becomes
colourless. Carry out a blank titration on a mixture of 10 mL Saponification value (2.5.6)
of the freshly prepared potassium iodide solution, 20 mL of Maximum 2.0.
water and 40 mL of hydrochloric acid. The difference between ASSAY
the titrations represents the amount of potassium iodate Gas chromatography (2.2.28): use the normalisation
required. Each mL of O.05M potassium iodate VS is equivalent procedure.
to 33.64 mg of C 17H,.BrN.
STORAGE examined in ethanol (96 per cen!! R and dilute to 10.0 mL
Strong Cetrimide Solution should be stored at a temperature with the same solvent.
above 15 0 • Reference solution (a) Dissolve 50 mg of cetyl alcohol CRS in
LABELLING ethanol (96 per cen!! R and dilute to 5 mL with the same
The label states whether Ethanol, Isopropyl Alcohol or both solvent.
are present and the percentage of cetrimide, weight in Reference solution (b) Dissolve 50 mg of stearyl alcohol R in
volume. ethanol (96 percent) R and dilute to 10 mL with the same
solvent.
Reference solution (c) Mix I mL of reference solution (a)
and I mL of reference solution (b) and dilute to 10 mL with
Cetyl Alcohol ethanol (96 per cenl) R.
Column:
(Ph. Bur. monograph 0540) - size: I = 30 m, 0 = 0.32 mm,
Action and use - stationary phase: melhylpolysiloxane R (I urn).
Excipient. Carrier gas helium for chronuuogrophy R.
PhE" _ Flow rate 1 mUmin.
Splir ratio 1:100.
DEFINITION
Temperature:
Mixture of solid alcohols, mainly hexadecan-I-ol (C1oHJ'O;
klr 242.4), of animal or vegetable origin.
Time Temperature
Content (mIn) CC)
Minimum 95.0 per cent of C1.sH340. Column 0-20 150 ..... 250
CHARACTERS 20 - 40 250
Appearance Injection port 250
White or almost white, unctuous mass, powder, flakes or Detector 250
granules.
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1-518 Cetyl Palmitate 2022
/lljution 1 ilL of the test solution and reference Heat under reflux for 2 h.
solutions (a) and (c). Alkaline impurities
System suitability Referencesolution (c): Dissolve 2.0 g with gentle heating in a mixture of 1.5 mL of
- resolution: minimum 5.0 between the peaksdue to cetyl ethanol (96 per e"n) Rand 3 mL of toluene R. Add 0.05 mL
alcohol and stearyl alcohol. of a 0.4 gIL solution of bromophenol blue R in ethanol
Calculate the percentage content of C 16H340. (96 per cenO R. Not more than 0.4 mL of 0.01 M hydrlXhlori<
________ ~ PIIE« acid is required to change the colour of the solution to
yellow.
Water (2.5.12)
Maximum 0.3 per cent, determined on 1.0 g using a mixture
Cetyl Palmitate of equal volumes of anhydrous methanol R and methylene
chloride R as solvent.
(Ph. Eur. monograph 1906) Total ash (2.4.16)
Action and use Maximum 0.2 per cent, determined on 1.0 g.
Excipient. ASSAY
PIIE« _ Gas chromatography (2.2.28): use the normalisation
procedure.
DEFINITION Test solution Dissolve 20.0 mg of the substance to be
.Mixture of esters of CH-Cra alcohols with lauric examined in hexane R and dilute to 20.0 mL with the same
(dodecanoic), myristic (terradecanoic), palmitic solvent.
(hexadecanoic) and stearic (octadecanoic) acids ('Celyl esters Reference solution (a) Dissolve 20.0 mg of
wax').
my! palmitate 95 CRS in hexane R and dilute to 20.0 mL
Content with the same solvent.
(expressed as hexadecyl hexadecanoare): Reference solution (b) Dissolve 20.0 mg of
10.0 per cent to 20.0 per cent for Cetyl palmitate 15, rety/ palmitate 15 CRS in hexane R and dilute to 20.0 mL
60.0 per cent to 70.0 per cent for Cetyl palmitate 65 and with the same solvent.
minimum 90.0 per cent for Cetyl palmitate 95. Column:
CHARACTERS - nuuesial: stainless steel;
Appearance . - size: 1= 10 m, 0 = 0.53 mm;
White or almost white, waxy plates, flakes or powder. - stationary phase: methylpo/ysiloxane R (film thickness
2.65 pm).
Solubility
Practically insoluble in water, soluble in boiling anhydrous Comer gas helium for ehromaUJgraphy R.
ethanol and in methylene chloride, slightly soluble in light Flow rate 6.5 mUmin.
petroleum,practically insoluble in anhydrous ethanol. Split ratio I: IO.
mp Temperature:
About 45 "C for Cetyl palmitate 15 and Cetyl palmitate 65
and about 52 "C for Cetyl palmitate 95. Tun, Temperature
(nUn) ("C)
IDENTIFICATION
Column 0-10 100 ~ 300
A. It complies with the limits of the assay and me
10 - ]5 300
chromatogram obtained with me test solution shows the
Injection port 350
typical main peak(s).
Detector 350
B. Saponification value (see Tests).
TESTS Detection Flame ionisation.
Appearance of solution Injection I ~L.
The solution is not more intensely coloured than reference
Relative retention With reference to ceryl palmitate (retention
solution Y. (2.2.2, Method II).
time = about 9 min): cetyl alcohol = about 0.3; palmitic
Dissolve 4.0 g in methylene chloride R and dilute'0
20 mL acid = about 0.4; lauric ester = about 0.8; myristic
with the same solvent. ester = about 0.9; stearic ester = about 1.1.
Acid value (2.5.1) System suitabi/ityReference solution (b):
Maximum 4.0. - resolution: minimum 1.5 between the peaks due to cetyl
Dissolve 10.0 g in 50 mL of the solvent mixture descnbed by palmitate and cetyl stearate.
heating under reflux on a water-bath for 5 min.
STORAGE
Hydroxyl value (2.5.3, MethodA) At a temperature not exceeding 25°C.
Maximum 20.0.
LABELLING
Carry out the titration at a temperature between 55°C and
The label states the type of cetyl palmitate.
70 °C, shaking the flask towards the end of the titration.
__________________ ~ PIIE«
Iodine value (2.5.4, Memod A)
Maximum 2.0.
105 to 120.
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2022 Chalk 1-519
curve shows no point of inflexion, the subsrance to be

Cetylpyridinium Chloride examined does not comply with the test. If the curve shows
(Ph. Eur. monograph 0379) one point of inflexion) repeat the test but add 3.0 mL of a
25.0 gIL solution of dimethyldecylamine R in 2-propanol R
before the titration.Uthe titration curve after the addition of
CH, 12.0 mL of the titrant shows only one point of inflexion, the
substance to be examined does not comply with the test.
Water (2.5.12)
C"H,.CIN,H,O 358.0 6004-24-6 4.5 per cent to 5.5 per cent, determined on 0.300 g by the
semi-micro determination of water.
Action and use Sulfated ash (2.4.14)
Antiseptic. Not more than 0.2 per cent, determined on 1.0 g.
PhE" _
ASSAY
DEFINITION Dissolve 2.00 g in water R and dilute to 100.0 mL with the
Cetylpyridinium chloride containsnot less than 96.0 per cent same solvent. Transfer 25.0 mL of the solution to a
and not more than the equivalent of 101.0 per cent of separating funnel, add 25 mL of chloroform R, 10 mL of
I-hexadecylpyridinium chloride, calculated with reference to 0.1 M sodium hydroxide and 10.0 mL of a freshly prepared
the anhydrous substance. 50 gIL solution of potassium iodide R. Shakewell, allow to
separate and discard the chloroform layer. Shake the aqueous
CHARACTERS layer with three quantities, each of 10 ml., of chlorofonn R
A white or almost white powder, slightly soapy to the touch, and discard the chloroform layers. To the aqueous layeradd
soluble in water and in alcohol. An aqueous solution froths 40 mL of hydrochlori< acid R, aUow to cool and titrate with
copiously when shaken. 0.05 M potassium iodate until the deep-brown colour is almost
IDENTIFICATION discharged. Add 2 mL of chloroform R and continue the
First identification: B, D. titration) shaking vigorously, until the chloroform layerno
longerchangescolour. Carry out a blanktitration on a
mixture of 10.0 mL of the freshly prepared 50 gIL solution
A. Dissolve 0.10 gin wa'" R and dilute to 100.0 mL with of potassium iodide R, 20 mL of water R and 40 mL of
the same solvent. Dilute 5.0 mL of this solution to 100.0 mL hydrochloric acid R.
with water R. Examined between 240 nm and 300 nm
1 mL of 0.05 M potassium iodate is equivalent to 34.0 mg of
(2.2.25), the solutionshows an absorption maximum at
259 om and 2 shoulders at about 254 om and at about C 21 H" CIN.
__ ~ ~ PhE"
265 nm. The specific absorbance at the maximum is 126 to
134, calculated with reference to the anhydrous substance.
B. Examine by infrared absorption spectrophotometry
cety!pyddinium chloride CRS. Examine the substances in the Chalk
solid state. Prepared Chalk
C. To 5 mL of dilute sodium hydroxide solutUm R add 0.1 mL CaCO, 100.1
of bromophenol blue solution Rl and 5 mL of chlcr%nn R and
shake. The chloroform layer is colourless. Add 0.1 mL of Action and use
solution S (see Tests) and shake. The chloroform layer Antacid.
becomes blue.
D. Solution S gives reaction (a) of chlorides (2.3.1). DEFINITION
Chalkis a native form of calcium carbonate freed from most
TESTS of its impurities by elutriation and dried. It containsnot less
Solution S than 97.0% and not more than 100.5% orCaC03 ,
Dissolve 1.0 g in carbon dioxide-free water R and dilute to calculated whh reference to the dried substance.
CHARACTERISTICS
Chalkabsorbs waterreadily.
suspension 11(2.2.1) and is colourless (2.2.2, Method II). Practically insoluble in water, slightly soluble in water
containing carbondioxide.
Acidity
To 50 mL of solution S add 0.1 mL of phenolphthalein Macrosropkal White or greyish white, smallfriable masses,
solution R. Not more than 2.5 mL of 0.02 M sodium hydroxide usually conical in form, or in powder; amorphous; earthy;
is required to change the colour of the indicator. soft to the touch.
1Wkroscopkal Consists of the calcareous shells and detritus
Amines and amine salts
of various foraminifera; the calcareous shells vary from about
Dissolve 5.0 g with heating in 20 mL of a mixture of
35 to 100 um in breadth and from about 50 to 180 um in
3 volumes of 1 M hydrochloric acid and 97 volumes of
length; among the detritus are numerous small rings and
methanol R and add 100 mL of 2-propanol R. Pass a stream
discs about 5 to 10 pm in diameter.
of nitrogen R slowly through the solution. GraduaUy add
12.0 mL of 0.1 M tetrabutylammonium hydroxide and IDENTIFICATION
record the potentiometric titration curve (2.2.20). lithe curve A. A solution in 6M QUlr'C add yields reaction C characteristic
shows 2 points of inflexion) the volume of titrant added of calcium salts, Appendix VI.
between the two points is not greater than 5.0 mL. If the B. Yieldsreaction A characteristic of carbonates, Appendix VI.
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1-520 Charcoal 2022
TESTS IDENTIFICATION
Acidity or alkalinity A. When heated to redness it bums slowly without a flame.
I g, boiled with 50 mL of water and filtered, yields a filtrate B. Adsorption power (see Tests).
which is neutralto bromothymol blue solution Rl or requires
not more than 0.05 mL of O.lM hydrochlori< acid VS to make TESTS
it so. Solution S
To 2.0 g in a conical flask with a ground-glass neck add
Aluminium, iron, phosphate and matter insoluble in 50 mL of dIlute hydrochlori< acid R. Boil gently under a reflux
hydrochloric acid condenser for 1 hi filter and wash the filter with dilute
Dissolve 2 g in a mixture of 5 mL of hydrochloric acid and hydrochloric acidR. Evaporate the combined filtrate and
75 mL of water, boil to remove carbon dioxide and make washings to dryness on a water-bath, dissolve the residue in
alkaline with 5M ammonia using methyl red solution as 0.1 M hydrochloric acid and dilute to 50.0 mL with the same
indicator. Boil for 1 minute, filter and wash the precipitate acid.
with a hot 2% w/v solution of ammonium chloride. Dissolve
the precipitate as completely as possible by passing 20 mL of Acidity or a1kalinlty
hot 2M hydrochloric acid through the filter and wash the filter To 2.0 g add 40 mL of water R and boil for 5 min. Cool,
with sufficient hot water to adjust the volume of the solution restore to the original mass with carbon dioxide-free water R
to 50 mL. Boil the solution and make alkaline with and filter. Reject the first 20 mL of the filtrate. To 10 mL of
5M ammonia using methyl red solution as indicator. Boil for the filtrate add 0.25 mL of bromolhymol blue solution R1 and
1 minute, filter through the same filter, wash the precipitate 0.25 mL of 0.02 M sodium hydroxide. The solution is blue.
with a hot 2% w/v solution of ammonium nitrate, dry and Not more than 0.75 mL of 0.02 M hydrochlori< add is
ignite at a temperature not lower than 10000 • The residue required to change the colour of the indicator to yellow.
weighs not more than 40 mg. Acid-soluble substances
Arsenic Maximum 3 per cent.
Dissolve 0.5 g in 5 mL of brominated hydrochloric acid and To 1.0 g add 25 mL of daul< nitric acid R and boil for 5 min.
dilute to 50 mL with water. 25 mL of the resulting solution Filter whilst hot through a sintered-glass filter (10) (2.1.2)
complies with the limit test for onenic, Appendix vn (4 ppm). and wash with 10 mL of hot water R. Evaporate the
Chloride combined filtrate and washings to dryness on a water-bath,
Dissolve 0.3 g in 2 mL of nitrk acidand 10 mL of water, add to the residue 1 mL of hydrrxhlon·c acidR, evaporate to
dryness again and dry the residue to constantmass at
filter and dilute the filtrate to 30 mL with water. 15 mL of
100-105 "C. The residue weighs a maximum of 30 mg.
the resulting solution complieswith the lim;" test/or chlorides,
Appendix vn (330 ppm). . Alkali-soluble coloured substances
To 0.25 g add 10 mL of dllul< sodium hydroxide solution R
Sulfate
Dissolve 0.25 g in 5.5 mL of 2M hydrochloric acid, dilute to and boil for 1 min. Cool, filter and dilute the filtrate to
10 mL with water R. The solution is not more intensely
30 mL with water and filter. 15 mL of the resulting solution
coloured than reference solution GY 4 (2.2.2, Muhod II).
complies with the limit test for sulfares, Appendix vn (0.12%).
Ethanol (96 per cent) soluble substances
Loss on drying
When dried to constant weightat 105°J loses not more than
1.0% of its weight. Use 1 g. To 2.0 g add 50 mL of ethanol (96 per cen!> R and boil under
a reflux condenser for 10 min. Filter immediately, cool, and
ASSAY dilute to 50 mL with ethanol (96 per cen!> R. The filtrate is
To 2 g in 100 mL of water add 50 mL of 1M hydrochlori< acid not more intensely coloured than reference solution Y6 or
VS, boil to remove carbon dioxide, cool and titrate the excess BY. (2.2.2, Melhod ll). Evaporate 40 mL of the filtrate to
of acid with 1M sodium hydroxide VS using muhyl orange dryness and dry to constant mass at 100-105 °C. The residue
solution as indicator. Each mL of 1M hydrochloric add VS is weighs a maximum of 8 mg.
equivalent to 50.04 mg of CaC03 •
Fluorescent substances
In an intermittent-extraction apparatus, treat 10.0 g with
100 mL of cycIohe:cane R1 for 2 h. Collect the liquid and
Activated Charcoal dilute to 100 mL with cydohexane R 1. Examine in ultraviolet
light at 365 nm. The fluorescence of the solution is not more
DecolourisingCharcoal intense than that of a solution of 83 J.lg of quinine R in
(Ph. Eur. monograph 0313) 1000 mL of o. 005 M sulfuric acid examined under the same
conditions.
Action and use Sulfides
Adsorbent. To 1.0 g in a conical flask add 5 mL of hydrochloric add R1
PhE<r ~_
and 20 mL of waterR. Heat to boiling. The fumes released
do not tum leadacetal< paper R brown.
DEFINITION Copper
Obtained from vegetable matter by suitable carbonisation Maximum 25 ppm.
processes intended to confera high adsorption power.
CHARACTERS Test solulion Use solution S.
Appearance Reference solutions Prepare the reference solutions using
Black, light powder free from grittiness.
copper standard solution (0.1 per cen' Cu) R and diluting with
Solubility 0.1 M hydrochloric acid.
Practically insoluble in aU usualsolvents.
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2022 Chenodeoxycholic Acid 1-521
*****
Chenodeoxycholic Acid
Wavelength 325.0 om. ** **
Atomisation device Air-acetylene flame. (Ph. Eur. monograph 1189) ***
Lead
Maximum 10 ppm.
Atomic absorption spectrometry (2.2.23, Method/). co,H
Test solution Use solution S.
Reference solutions Prepare the reference solutions using lead
standardsolution (100 ppm Pb) R and diluting with 0.1 M
hydrochloric acid.
Source Lead hollow-cathode lamp.
Wavelength 283.3 nm; 217.0 urn may be used depending 392.6 474-25-9
on the apparatus.
Action and use
Atomisation device Air-acetylene flame. Bile acid; treatment of gallstones.
Zinc
""Ell _
Maximum 25 ppm.
Atomic absorption spectrometry (2.2.23, Method 1). DEFlNITION
Testsolution Use solution S. Chenodeoxycholic acid contains not less than 99.0 per cent
Reference solutions Prepare the reference solutions using zinc and not more than the equivalent of 101.0 per cent of 3iX,7(/.-
standardsolution (100 ppm Zn) R and diluting with 0.1 M dihydroxy-5j3-cholan-24-oic acid, calculated with reference to
hydrochloric acid. the driedsubstance.
Source Zinc hollow-cathode lamp. CHARACTERS
Wavelength 214.0 urn. A white or almost whitepowder, very slightly solublein
water, freely solublein alcohol, soluble in acetone, slightly
soluble in methylene chloride.
Maximum 15 per cent, determined on 1.00 g by drying in an IDENTIFICATION
oven at 120 "C for 4 h. First identification: A.
Sulfated ash (2.4.14) Second identification: B, C.
Maximum 5.0 per cent, determinedon 1.0 g. A. Examine by infrared absorption spectrophotometry
Adsorption power
chenodeoxycholic add CRS. Examine the substances prepared
To 0.300 g in a .l00 mL ground-glass-stoppered conical flask
as discs using potassium bromide R.
add 25.0 mL of a freshly prepared solution of 0.5 g of
phenazone R in 50 mL of waterR. Shake thoroughly for B. Examine the chromatograms obtained in the test for
15 min. Filter and rejectthe first 5 mL of filtrate. related substances. The principal spot in the chromatogram
To 10.0 mL of the filtrate add 1.0 g of potassium bromide R obtained with test solution (b) is similar in position, colour
and 20 mL of dilute hydrochloric acid R. Using 0.1 mL of and size to the principal spot in the chromatogram obtained
methyl red solution R as indicator, titrate with 0.0167 M with reference solution (a).
potassium bromate until the red colour is discharged. Titrate C. Dissolve about 10 mg in I mL of sulfuric acidR.
slowly (1 drop every 15 s) towards the end of the titration. Add 0.1 mL offormaldehyde so/ution Rand aUow to stand foe
Carry our a blank titration using 10.0 mL of me phenazone 5 min. Add 5 mL of water R. The suspension obtained is
solution. greenish-blue.
Calculate the quantity of phenazone adsorbed per 100 g of TESTS
activated charcoal from the following expression: Specific optical rotation (2.2.7)
2.353(a - b) the same solvent. The specific optical rotation is + 11.0 to
m + 13.0, calculated with reference to the dried substance.
a number of millilitres of 0.0167 M potassiumbromate used for the Related substances
r blank; Examine by thin-layer chromatography (2.2.27), using a
b number of millilirres or 0.0167 M pOlossilDlJ bromau used (or the suitable silica gel as the coating substance.
test;
m mass in gramsof the substance to be examined. Test solution (aJ Dissolve0.40 g of the substance to be
examined in a mixture of 1 volumeof water Rand 9 volumes
Minimum 40 g of phenazoneis adsorbed per 100 g of of acetone R and dilute to 10 mL with the same mixture of
activated charcoal, calculated with reference to the dried solvents.
substance. Testsohuion (b) Dilute 1 mL of test solution (a) to 10 mL
Microbial contamination with a mixture of I volumeof water Rand 9 volumes of
TAMC: acceptance criterion 10' CFUig (2.6.12). acetone R.
TYMC: acceptance criterion 10' CFUlg (2.6.12). Reference sohuion (aJ Dissolve 40 mg of chenodeoxycholic
acid CRS in a mixture of 1 volume of water Rand 9 volumes
STORAGE
of acetone R and dilute to 10 mL with the same mixture of
In an airtight container. solvents.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""Ell
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1-522 Chenodeoxycholic Acid 2022
Reference solution (b) Dissolve 20 mg of lithocholic acidCRS IMPURITIES

in a mixture of 1 volume of waur Rand 9 volumes of
acetone R and dilute to 10 mL with the same mixture of
solvents. Dilute 2 mL of the solution to 100 mL with a
mixture of 1 volume of water Rand 9 volumes of acetone R.
Reference solution (c) Dissolve 20 mg of vnodeoxycbolic
acid CRS in a mixture of 1 volume of water Rand 9 volumes
of acetone R and dilute to 50 mL with the same mixture of
solvents.
Reference solution (d) Dissolve 20 mg of choltc acid CRS in a
mixture of 1 volume of water Rand 9 volumes of acetone R A. 3~-7Jl-dihydroxy-5p-cholan-24-oic acid (ursodeoxycholic
and dilute to 100 mL with the same mixture of solvents. acid),
Reference solution (e) Dilute 0.5 mL of test solution (a) to
20 mL with a mixture of 1 volume of water R and 9 volumes
of acetone R. Dilute 1 mL of the solution to 10 mL with a co,H
mixture of I volume of water Rand 9 volumes of acetone R.
Reference solution (f) Dissolve 10 mg of cheno<koxycholic
acid CRS in reference solution (c) and dilute to 25 mL with
the same solution.
Apply separately to the plate 5 IJL of each solution. Develop
in an unsaturated tank over a path of 15 em using a mixture B. 3~,7a,12a-trihydroxy-5p-cholan-24-oic acid (cholic acid),
of 1 volume of glacial acetic add R J 30 volwnes of acetone R
and 60 volumes of methylene chloride R. DIY the plate at
120 DC for 10 min. Spray the plate immediately with a
co,H
47.6 gIL solution of phosphomolybdic acidR in a mixture of
1 volume of sulfim·, add R and 20 volumes of glacial acetic
acid R and heat again at 120 DC until blue spots appear on a
lighter background. In the chromatogram obtained with lest
solution (a): any spot corresponding to lithocholic acid is not
more intense than the principal spot in the chromatogram
obtained with reference solution (b) (0. t per cent); any spot C. 3a-hydroxy-5p-cholan-24-oic acid (lithocholic acid),
corresponding to ursodeoxycholic acid is not more intense
than the principal spot in the chromatogram obtained with
reference solution (c) (1 per cent); any spot corresponding to
CO,"
cholic acid is not more intense than the principal spot in the
chromatogram obtained with reference solution (d)
(0.5 per cent); any spot apart from the principal spot and any
spots corresponding to lithocholic acid, ursodeoxycholic acid
and cholic acid, is not more intense than the principal spot in
(0.25 per cent). The test is not valid unless the
D. 3~, 7p,12a-trihydroXY-5p-cholan-24-oic acid (ursochclic
chromatogram obtained with reference solution (f) shows two
acid),
clearly separated principal spots.
CD,H
drying in an oven at 105 DC.
Sulfated ash (2.4.1</)
ASSAY
Dissolve 0.350 g in 50 mL of alcohol R, previously
neutralised to 0.2 mL of phenolphlhalein solution R.
E. 3~,12~-dihydroxy-5Jl-cholan-24-oic acid (deoxycholic
Add 50 mL of waterR and titrate with 0.1 M sodium
acid),
hydroxUk until a pink colour is obtained.
C24HI004.
CD,H
F. 3a-hydroxy-7-oxo-5p-cholan-24-oic acid,
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2022 Chitosan Hydrochloride 1-523
with water and dry to constant weight in an oven at

100-105 °C. The residue weighs a maximum of 10 mg.
pH (2.2.3)
4.0 (0 6.0 for solution S.
Viscosity (2.2.11J)
80 per cent to 120 per cent of the value stated on the label,
determined on solution S.
G. methyl 3,,7~-dihydroxy-5~-cholan-24-oate. Determine the viscosity using a rotating viscometer at 20 °C
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'" with a spindle rotating at 20 rlmin, using a suitable spindle
for the range of the expected viscosity.
Degree of deacetylation
Testsolution Dissolve 0.250 g in water R and dilute to
Chitosan Hydrochloride ***
*** ***
50.0 mL with the same solvent, stirring vigorously. Dilute
1.0 mL of this solution to 100.0 mL with water R. Measure
(Ph. Eur. monograph 1774) *** the absorbance (2.2.25) from 200 run to 205 run as the first
PhE" _ derivative of me absorbance curve. Determine me pH of the
solution.
DEFINITION Reference solutions Prepare solutions of 1.0 ~glrnL,
Chitosan hydrochloride is the chloride salt of an unbranched 5.0 ~glrnL, 15.0 ug/ml, and 35.0 ~glrnL of N-
binary heteropolysaccharide consisting of the two units acetylglucosarmne R in water R. Measure the absorbance
N-acetyl-D-glucosamine and n-glucosamine, obtained by (2.2.25) from 200 run to 205 run of each solution as the first
partial deacetylation of chitin normally leading to a degree of derivative of the absorption curve. Make a standard curve by
deacetylation of 70.0 per cent to 95.0 per cent. Chitin is plotting the first derivative at 202 om as a function of me
extracted from the shells of shrimp and crab. concentration of N-acetylglucosamine, and calculate the slope
PRODUCTION of the curve by least squares linear regression. Use the
The animals from which chitosan hydrochloride is derived standard curve to determine tile equivalent amount of
must fulfil the requirements for the health of animals suitable N-acetylglucosamine for the substance to be examined.
for human consumption to the satisfaction of the competent Calculate the degree of deacetylation (molar) using me
authority. It must have been shown to what extent the following expression:
method of production allows inactivation or removal of any
contamination by viruses or other infectious agents. lOOxM,x(C,-C2 )
(M, xC,) - [(M, - M,) x C21
CHARACTERS
Appearance C1 concentration of chitosan hydrochloride in the lest solution in
White or almost white, fine powder. micrograms per miUilitrej
Cz concentration of N-acetylglucosamine in the test solution. as
Solubility determined from the standard curve prepared using the
Sparingly soluble in water, practically insoluble in anhydrous reference solution in micrograms per miHilit:re;
ethanol. M1 203 (reJarive molecular mass of N-ace!ylglucosamine unit
(CsH 13 N O j ) in polymer);
IDENTIFICATION MJ relative molecular mass of chitosan hydrochloride.
Preparation Discs. M J is calculated from the pH in solution, assuming a pKa
value of 6.8, using the following equations:
Comparison chiwsan hydrochloride CRS.
B. It gives reaction (a) of chlorides (2.3.1). M, =fxM2 +(I-f)x (M 2 +36.5)
C. Dilute 50 mL of solution S (see Tests) to 250 mL with a
25 per cent VIV solution of ammonia R. A voluminous P
gelatinous mass is fonned. f=l+P
D. To 10 rnL of solution S add 90 rnL of acetone R.
A volwninous gelatinous mass is formed. p = 10(PH-pK.)
TESTS
Ml 161 (relative molecular mass ofdeacetylated unit (gIucosamine)
Soludon S (CJ-lllN04) in polymer).
Dissolve 1.0 g in 100 mL of water R and stir vigorously for
20 min with a mechanical stirrer. Chlorides
Appearance of solution 10.0 per cent to 20.0 per cent.
Solution S is not more opalescent than reference Introduce 0.200 g into a 250 mL borosilicate flask fitted with
suspension n (2.2.1) and not more intensely coloured than a reflux condenser. Add 40 mL of a mixture of I volume of
reference solution BY, (2.2.2, Method II). niaic acid Rand 2 volumes of water R. Boil gently under a
Matter insoluble in water reflux condenser for 5 min. Cool and add 25 mL of waterR
Maximum 0.5 per cent. through the condenser. Add 16.0 rnL of 0.1 M savernitrate,
Add 2.00 g to 400.0 rnL of water R while stirring until no shake vigorously and titrate with 0.1 M" ammonium
further dissolution takes place. Transfer the solution to a 2 L thiocyanate, using ] rnL of feme ammonium sulfate solution R2
beaker, and add 200 mL of water R. Boil the solution gently as indicator, and shaking vigorously towards the end-point.
for 2 h, covering the beaker during the operation. Filter Carry out a blank titration.
through a sintered-glass filter (40) (2.1.2), wash the residue 1 mL of 0.1 wI silver nitrate is equivalent to 3.55 mg of CI.
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1-524 Chloral Hydrate 2022
Loss on drying (2.2.32) Chlorides (2.4.4)

Maximum 10 per cent, determined on 1.000 g by drying in Maximum 100 ppm.
an oven at 105°C. Dilute 5 mL of solution S to 15 mL with water R.
Sulfated ash (2.4.14) Non-volatile residue
Maximum 1.0 per cent, determined on 1.0 g. Maximwn 0.1 per cent.
STORAGE Evaporate 2.000 g on a water-bath. The residue weighs a
At a temperature of 2 °C to 8°C, protected from moisture maximum of 2 mg.
and light. ASSAY
LABELLING Dissolve 4.000 g in 10 mL of water R and add 40.0 mL of
The label states the nominal viscosity in millipascal seconds 1 M sodium hydroxide. Allow to stand for exactly 2 min and
for a 10 gIL solution in water R. titrate with 0.5 M sulfuric acid, using 0.1 mL of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEw phenolphthalein solution R as indicator. Titrate the neutralised
solution with 0.1 M silver niuxue, using 0.2 mL of potassium
chromate solurion R as indicator. Calculate the number of
millilitres of 1 M sodium hydroxide used by deducting from
the volume of 1 M sodium hydroxide, added at the beginning
Chloral Hydrate of the titration) the volume of 0.5 M sulfuric addused in the
1S[ titration and two-fifteenths of the volwne of 0.1 M silver
nitrate used in me 2nd titration.
I mL of 1 M sodium hydroxide is equivalent to 0.1654 g
of (;,H,CI,O,.
STORAGE
(;,H,CI,O, 165.4 302-17-0 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'hE"
Action and use

Hypnotic.
Preparation
Chloral Hydrate Oral Solution Chlorambucil
PhE" _ (ph. Eur. monograph 0137)
DEFINITION
~CO,H
-
2,2,2-Trichloroethane-l ,I-diol.
Content
Cl~NA,.)I
CHARACTERS CI
Appearance
Colourless, transparent crystals. 304.2 305-03-3
Solubllity
Verysoluble in water, freely soluble in ethanol (96 per cent). Action and use
Cytotoxicalkylating agent.
IDENTlFICATlON
Preparation
A. To 10 mL of solution S (see Tests) add 2 mL of dilute
Chlorambucil Tablets
sodium hydroxide solution R. The mixture becomes cloudy
and, when heated, gives off an odour of chloroform. PhE" _
B. To I mL of solution S add 2 mL of sodium sulfide
DEFINITION
solution R. A yellow colour develops whichquickly becomes
4-[4-[Bis(2-chloroethyl)amino]phenyl]butaooic acid.
reddish-brown. On standing for a short time, a red
precipitate may be formed. Content
TESTS
Soludon S CHARACTERS
Dissolve 3.0 g in carbor, dioxide-free waterR and dilute to Appearance
30 mL with the same solvent. White or almost white) crystalline powder.
Appearance of solution Solubllity
Solution S is clear (2.2.1) and colourless (2.2.2, Methodll). Practically insoluble in water, freely soluble in acetone and in
ethaool (96 per cent).
pH (2.2.3)
3.5 to 5.5 for solution S. IDENTIFICATION
Chloral alcoholate Infrared absorption spectrophotometry (2.2.24).
Warm 1.0 g with 10 mL of dilu" sodium hydroxide solution R, Comparison chlorambucil CRS.
filter the supernatant solution and add 0.051"/ iodine
dropwise until a yellow colour is obtained. Allow to stand for
1 h. No precipitate is formed.
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2022 Chlorambucil 1-525
TESTS Flow rate 0.8 mUmin.

Impurity G Detection Spectrophotometer at 260 om.
Liquid chromatography (2.2.2'l). The solutions are stable for Injection 10 ~L.
8 h at room temperature orfor 24 h at 4-8 DC; protect them from
light.
with chlorambudl for system suitability CRS and the
Test solution Dissolve 10 mg of the substance to be chromatogram obtained with reference solution (b) to
examined in methanol R and dilute to 20.0 mL with the same identify the peaks due to impurities Band E.
solvent.
Relative retention With reference to chlorambucil (retention
Reference solution (aJ Dilute 1.0 mL of the test solution to time == about 12 min): impurity B = about 0.5;
50.0 mL with the mobile phase. Dilute 2.0 mL or this
solution to 10.0 mL with me mobile phase.
=
System suitabl1ity Reference solution (b):
Reference solutio" (b) Dissolve 5 mg of chlorambucil with - resolution: minimum 5.0 between the peaks due to
impurity G CRS in methanol R and dilute to 10.0 mL with impurity B and chlorambucil.
the same solvent.
Limits:
Column: - impurity E: not more than 6 times the area of the principal
- size: /;;: 0.15 m, 0 :::: 3.9 mm; peak in the chromatogram obtained with reference
- stationary phase: phmylsilylsHica gelfor chromatography R solution (a) (0.6 per cent);
(5 1lJl1). - impun·ty B: not more than 4 times the area of the
Mobile phase methanolR, 1 per cent VIV solution of principal peak in the chromatogram obtained with
trilluoroocetic acidR (50:50 VIV). reference solution (a) (0.4 per cent);
Flow rate 1.8 mUmin. - unspecified impurities: for each impurityJ not more than the
Detection Spectrophotometer at 260 run. area of the principal peak in the chromatogram obtained
Injeaion 20 ~L. - total: not more than 10 times the area of the principal
Run time Twice the retention time of chlorambucil. peak in me chromatogram obtained with reference
Relative retention With reference to chlorambucil (retention solution (a) (1.0 per cent);
time > about II min): impurity G ::::: about 1.2. - disregard limit". 0.5 times the area of me principal peak in
System suitability Reference solution (b): the chromatogram obtained with reference solution (a)
- resolution: minimum 1.5 between the peaks due to (0.05 per cent).
chlorambucil and impurity G. Water (2.5.12)
Limit: Maximum 0.5 per cent, determined on 1.00 g.
- impun·ty G: not more than the area of the principal peak SuIrated ash (2.4.14)
in the chromatogram obtained with reference solution (a) Maximum 0.1 per cent, determined on 1.0 g.
(0.4 pet cent).
ASSAY
Related substances Dissolve 0.200 g in 10 mL of acetone R and add 10 mL or
Liquid cbsomatography (2.2.29). Prepare the solutions
water R. Titrate with 0.1 M sodium hydroxide, using 0.1 mL
immediately before use and protect from light of phenolphthalein solution R as indicator.
Solvent mixture 10.3 gIL solution of hydrochlori< acidR,
acetoniuile for chromatography R (10:90 VIV).
C 14H1.CI2N02 •
examined in the solvent mixture and dilute to 100.0 mL with STORAGE
the solvent mixture. Protected from light.
Reference solution (a) Dilute 1.0 mL of the test solution to IMPURITIES
100.0 mL with the solvent mixture. Dilute 1.0 mL or this Specified impurities B, E, G.
solution to 10.0 mL with the solvent mixture. Otherdetectable impurities (thefollowing substances would, if
Reference solution (b) Dissolve 5 mg or chlorambucil for system present at a safficiem level, be detected by oneor other of the tests
suitability CRS (containing impurities B and E) in the solvent in the monograph. They arelimited by the general aaeptatue
mixture and dilute to 20.0 mL with the solvent mixture. cruetion for other/unspecified impurities and/or by thegeneral
- size: 1= 0.25 m, 0 = 3.0 mm, therefore not necessary to idenu'fy these impun·lies for
- stationary phase: end-eapped octadtXYlsilyl silica gelfor demonstration ofcompliance. See also 5.10. Control of impun·lies
chromatography R (5 um). in substances for pharmaceutical uu) A, CJ D, F.
Mobilephase:
- mobile phase A: 1.9 gIL solution of ammonium tUetale R
adjusted to pH 3.9 with acetic acidR,
- mobile phaseB: acetonitrile for chromatography R;

0- 5 60 40 A. 4-[4-[(2-chloroethyl)(2-hydtoxyethyl)amino]phenyI]
5· 15 60-1'10 40 ..... 90 butanoic acid,
15 - 25 10 90
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1-526 Chloramphenicol 2022
~co,H
CI <:»"> N J V
Chloramphenicol
H
B. 4-[4-[(2-chloroethyl)amino]phenyl)butanoic acid, OH NO,
CI~O
CI~o"'P",O~N)V
~C~H
o
CI~ ..-IN'.
r -H W
(H
H OH
'"
#
~ (
CI
56-75-7
Action and use

C.4-[4-[[2-[[bis(2-chloroethoxy)phosphoryl]oxy]ethyl](2- Antibacterial.
chloroethyl)amino]phenyl)butanoic acid,
Preparations
Chloramphenicol Capsules
~O~CI
Chloramphenicol Ear Drops
CI~N~ (,
Chloramphenicol Eye Drops
CI
( Chloramphenicol Eye Ointment
PhE" _~ ~ _
D. 2-chloroethyl 4-[4-[bis(2-chloroethyl)amino] DEFINITION

phenyllbutancate, 2,2-Dichloro-N-[(IR,2R)-1.3-dihydroxy-I-(4-nitrophenyl)
propan-2-yl]acetarnide.
Content
CHARACTERS
Appearance
White, greyish-white or yellowish-white, fine, crystalline
powder or fine crystals, needles or elongated plates.
Solubility
Slightly soluble in warer, freely soluble in ethanol
(96 per cent) and in propylene glycol.
E. 4-[4- [[2-[[4-[4-[bis(2-chloroethyl)amino]phenyl] bu tanoyl] A solution in anhydrous ethanol is dextrorotatory and a
oxy]ethyl](2-chloroethyl)amino]phenyl]butanoic acid, solution in ethyl acetate is laevorotatory.
IDENflFICATION
C(i
o,H
First identificatWn: A.
Second identification: B.
""I
~ N~O~fCl A. Infrared absorption spectrophotometry (2.2.24).
~ 8 ~N Comparison chlorampheni<ol CRS.

B. Melting poinr (2.2.14).
CI~O.J Determination A Determine the melting point of the
CI 1 0 substance to be examined.
~N
Results A 149 'C to 153 'C.
Determination B Mix equalparts of the substance to be
•
Cl examined and chloramphenicol CRS and determine the
melting point of the mixture.
F. 4-[4-[[2-[[4-[4-[[2-[[4-[4-[bis(2-chloroethyl) Results B The absolute difference between the melting point
amino ]phenyl] butanoyl] oxy]ethyl] (2-chloroethyl) of the mixture and the value obtained in determination A is
amino]phenyl)butanoyl]oxy]ethyl] (2-chloroethyl) not greater than 2 "C.
amino]phenyl)butanoic acid,
TESTS
r"") To 0.1 g add 20 mL of carbon dioxide-free water R, shake and
C1 ............... N~C02H add 0.1 mL of bromothymol blue solution Rl. Not more than
0.1 mL of 0.02 M hydrochlmi< add or 0.02 M sodium
hydroxide is required to change the colour of the indicator.
•
Cl
+ 18.5 to + 20.5.
G. 4-[2-[bis(2-chloroethyl)amino]phenyl]butanoic acid or
Dissolve 1.50 g in anhydrous ethanol R and dilute to 25.0 mL
4-[3-[bis(2-chloroethyl)amino]phenyl)butanoic acid (meta
or ortho chlorambucil).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
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2022 Chloramphenicol 1-527
Related substances - unspecified impurities: for each impurity, maximum

Liquid chromatography (2.2.29). Prepare the solutions 0.10 per cent;
immediately before use. - total: maximum 0.5 per cent;
Solution A Dissolve 2.0 g of sodium heptanesulfonate R in - reporting threshold: 0.05 per cent.
900 mL of waterfor chromatography R. Add 6.8 g of potassium Chlorides (2.4.4)
dihydrogen phosphate Rand 5 mL of triethylamine R. Adjust to Maximum 100 ppm.
pH 2.5 with phosphoric acid R and dilute to 1000 mL with To 1.00 g add 10 mL of niuic acid Rand 20 mL of waterR
waterfor chromatography R. and shake for 5 min. Filter through a filter paper previously
Test solution (a) Dissolve 20.0 mg of the substance to be washed by filtering 5 mL portions of water R until 5 mL of
examined in 10 mL of methanol R and dilute to 200.0 mL filtrate no longer becomes opalescent on addition of 0.1 mL
with mobile phase A. of nitric add Rand 0.1 mL of silver nitrate solution Rl. 15 mL
Testsolution (b) Dissolve 25.0 mg of the substance to be of me filtrate complies with the test.
examined in 5 mL of methanol Rand dilute to 50.0 mL with Loss on drying (2.2.32)
mobile phase A. Maximum 0.5 per cent, determined on 1.000 g by drying in
Reference solution (a) Dissolve 20.0 mg of an oven at 105°C.
chloramphenicol CRS in 10 mL of methanol R and dilute to Sulfated ash (2.4.14)
200.0 mL with mobile phase A. Maximum 0.1 per cent, determined on 2.0 g.
ASSAY
solution to 10.0 mL with mobile phase A. Liquid chromatography (2.2.29) as described in the test for
Reference solution (c) Dissolve 12.5 mg of
4-nitrobenzaidehyde R (impurity B) in 2 mL of methanol Rand Mob,le phase Mobile phase A.
dilute to 50 mL with mobile phase A. Injection Test solution (a) and reference solution (a).
Reference solution (d) Dissolve 5 mg of chloramphenicol for Run time 1.5 times the reter:ttion time of chloramphenicol.
peak identification CRS (containing impurity A) in 1 mL of Calculate the percentage content of CllHIZClzNzOj taking
methanol R, add 1 mL of reference solution (c) and dilute to into account the assigned content of chloramphenicol CRS.
10 mL with mobile phase A. STORAGE
Column: Protectedfrom light. If the substance is sterile, the container
- size: 1= 0.25 m, 0 = 4.6 mm; is also sterile, airtight and tamper-evident.
- stationary phase: base-deactivated end-capped ocutdecy/siiyl
silica gelfor chromatography R (5 pm); IMPURITIES
- temperature: 25°C. Spedfied impurities A.
Mobile phase: Otherdetectable impurities (thefollowing substances would, if
- mobile phaseA: methanol R, solution A (32:68 VII'); present at a sufficient level, be detected by one or other of the tests
- mobile phaseB: methanol R; in the monograph. Theyare limited by thegeneral acceptance
criterion for otherhmspedfied impurities and/or by thegeneral
Tim. Mobile phase A MobUe phase B monograph Substan«s for pharmaceutical use (2034). It is
(min) (per cent VIJI) (per cent V/J.? therefore not necessary to identify these impurities for
0-13 100 0 demonstration of compliance. See also 5.10. Control of impuri,ties
13 - 25 100 -. 60 0-.40 in substances for phannaceutical use) B.
25 - 33 60 40
"
OH
W
NO,
Flow rate 1.0 mUmin. ", H I
H,N , "'"
Detection Spectrophotometer at 277 run. HI OH
In;jecuOn 10 I!L of test solution (b) and reference
solutions (b) and (d).
A. (IR,2R)-2-amino-I-(4-nitrophenyl)propane-I,3-diol,
Identification of impuriues Use the chromatogram supplied
with chloramphenicol for peak idenu"fication CRS and the
Relative retention With reference to chloramphenicol
(retention time = about 14 min): impurity A = about 0.7; B. 4-nitrobenzaldehyde.
impurity B = about 0.9. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEII
impurity B and chloramphenicol.
Cakulation of percentage contents;
- correction factor. multiply the peak area of impurity A by
0.7;
chloramphenicol in reference solution (b).
Limits:
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1-528 Chloramphenicol Palmitate 2022
Chloramphenicol Palmitate *** 2 min and add 1 g of urea R, 2 mL of strong sodium hydroxide
*** *** solution R and 1 mL of p-naphthol solution R. A red colour
(Ph. Eur. monograph 0473) *** develops.
o TESTS
Acidity
Dissolve 1.0 g in 5 mL of a mixture of equal volumes of
ethanol (96 per cen!> R and ether R, warming to 35 'C.
CH,
Add 0.2 mL of phenolphthalein solution R. Not more than
0.4 mL of 0.1 M sodium hydroxide is required to produce a
pink colour persisting for 30 s.
561.6 530-43-8 Specific optical rotation (2.2.7)
Dissolve 1.25 g in anhydrous elhanol R and dilute to 25.0 mL
Action and use with the same solvent. The specific optical rotation is
Antibacterial. + 22.5 to + 25.5.
PhE" _ Free chloramphenicol
Maximum 450 ppm. Dissolve 1.0 g, with gentle heating) in
DEFINITION 80 mL of xylene R. Cool and shake with 3 quantities) each of
Chloramphenicol palmitate contains not less than 15 mL, of water R. Dilute the combined aqueous extracts to
98.0 per cent and not more than the equivalent of 50 mL with waterR and shake with 10 mL of toluene R.
102.0 per cent of (2R,3R)-2-[(dichloroaceryl)amino]-3- Allow to separate and discard the toluene layer. Centrifuge a
hydroxy-3-(4-nitrophenyl)propyl hexadecanoate, calculated portion of the aqueous layer and measure the absorbance (A)
with reference to the dried substance. (2.2.25) at the maximum at 218 om using as the
Semi-synthetic product derived from a fermentation product. compensation liquid a blank solution having an absorbance
not greater than 0.05.
CHARACTERS
A white or almost white, fine, unctuous powder, practically Calculate the content of free chloramphenicol in parts per
insoluble in water, freely soluble in acetone, sparingly soluble million from the expression:
in ethanol (96 per cent), very slightly soluble in hexane.
AxlO'
It melts at 87°C to 95 "C,
5.96
It shows polymorphism (5.9). The thennodyoamicaUy stable
form has low bioavailability following oral administration. Related substances
IDENTIFICATION Examine by thin-layer chromatography (2.2.27), using siJka
A. Examine by thin-layer chromatography (2.2.27), using gel GF254 R as the coating substance.
TLC silonised silica gelplate R. Testsolution Dissolve 0.1 g of the substance to be examined
Testsolution Dissolve 50 mg of the substance to be in acetone R and dilute to 10 mL with the same solvent.
examined in a mixture of 1 mL of 1 M sodium hydroxide and Reference solution (a) Dissolve 20 mg of chloramphenicd
5 mL of acetone R and allow to stand for 30 min. palmitate isomer CRS in acetone R and dilute to 10 mL with
Add 1.1 mL of 1 M hydrochloric acid and 3 mL of cuetone R. the same solvent. Dilute 1 mL of this solution to 10 rnL with
Reference solution (a) Dissolve 10 mg of chloramphenicd CRS acetone R.
in acewne R and dilute to 5 mL with the same solvent. Reference solution (b) Dissolve 20 mg of chloramphenkol
Reference solution (b) Dissolve 10 mg of palmi.c acidR in dipalmiune CRS in acetone R and dilute to 10 mL with the
acewne R and dilute to 5 mL with the same solvent. same solvent. Dilute 1 mL of this solution to 10 mL with
acetOne R.
Reference soaaion (c) Dissolve 10 mg of the substance to be
examined in acewne R and dilute to 5 mL with the same Reference solution (c) Dissolve '5 mg of chloramphenkol CRS
solvent. in auwne R and dilute to 10 mL with the same solvent.
Dilute 1 mL of this solution to 10 mL with acetone R.
Apply to the plate 4 J1L of each solution. Develop over a
path of 15 cm using a mixture of 30 volwnes of a 100 gIL Apply to the plate 10 J.lL of each solution. Develop over a
solution of ammonium acetate Rand 70 volumes of ethanol path of 15 ern using a mixture of 10 volumes of methanol R,
(96 per cen!> R. Allow the plate to dry in air and spray with a 40 volumes of chloroform Rand 50 volumes of <ycIohexane R.
solution containing 0.2 gIL of dichlorofluorescein R and Allow the plate to dry in air and examine in ultraviolet light
0.1 gIL of rhodamine B R in elhanol (96 per cent) R. Allow the at 254 run. In the chromatogram obtained with the test
plate to dry in air and examine in ultraviolet light at 254 nm, solution, any spots due to chloramphenicol palmitate isomer
The chromatogram obtained with the test solution shows and Chloramphenicol dipalmitate are not more intense than
3 spots corresponding in position to the principal spots in the the corresponding spots in the chromatograms obtained with
chromatograms obtained with reference solutions (a), (b) and reference solutions (a) and (b) respectively (2.0 per cent) and
(c). any spot, apart from the principal spot and the Spots due to
chloramphenicol palmitate isomer and chloramphenicol
B. Dissolve 0.2 g in 2 mL of pyridine R, add 2 mL of a
dipalmitate, is not more intense than the principal spot in the
100 gIL solution of potassium hydroxide R and heat on a
water-bath. A red colour is produced.
(0.5 per cent),
C. Dissolve about 10 mg in 5 mL of ethanol (96 per cen!> R
and add 4.5 mL of dilute sulfuric acid Rand 50 mg of zinc
powder R. Allow to stand for 10 min and if necessary decant
vacuo at 80°C at a pressure not exceeding 0.1 kPa for 3 h.
the supernatant or filter. Cool the solution in iced water and
add 0.5 mL of sodium nitrite solution R. Allow to stand for
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2022 Chloramphenicol Sodium Succinate 1-529
Sulfated ash (2.4.14) [(dichloroacetyl)amino]-3-hydroxy-I-(4-nitrophenyl)propyl

Maximum 0.1 per cent) derennined on 1.0 g. butanedioate (l isomer).
ASSAY Semi-syntheticproduct derived from a fermentation product.
Dissolve 90.0 mg in ethanol (96 percenO R and dilute to Content
100.0 mL with the same solvent. Dilute 10.0 mL of this 98.0 per cent to 102.0 per cent (anhydrous substance).
solution to 250.0 mL with ethanol (96 percenO R. Measure
CHARACTERS
the absorbance (2.2.25) of the solution at the maximum at
Appearance
271 urn.
White or yellowish-white powder, hygroscopic.
Calculate the content of C27H42ClzN206taking the specific
Solubility
absorbance to be 178.
Verysoluble in water, freely soluble in ethanol (96 per cent).
STORAGE
IDENTIFICATION
IMPURITIES Testsolution Dissolve 20 mg of the substance to be
examined in 2 ml, of aUWHe R.
OH N0:2
Reference solution (oJ Dissolve 20 mg of chloramphenicol
CI....
Y'~
°SZP···HI"'
JL
.
. " ..# sodium succinate CRS in 2 mL of acetone R.
Reference solution (b) Dissolve 20 mg of chlorampheni<oI CRS
CH, .CI H 0
in 2 mL of aawne R.
o Piau TLC silica gel GF254 plate R.
A. (IR,2R)-2- [(dichloroacetyl)amino]-3-hydtoxy-I-(4- Mobile phase dilute acetic acid R, methanol R, chlor%nn R
nitrophenyl)propyl hexadecanoate (chloramphenicol (1:14:85 VIVW).
palmitate isomer), Application 2 ~L.
Development Over a path of 15 cm.
o
szp
.
Drying In air.
CH, Detection Examine in ultraviolet light at 254 run.
o oH "'" NO,
Results The 2 principal spots in the chromatogram obtained
CI ... ~. ,#
Y'~ . with the test solution are similar in position and size to the
CH, CI H 0 2 principal spots in the chromatogram obtained with
reference solution (a); their positions are different from that
o of me principal spot in the chromatogram obtained with
B. (I R,2R) -2-[(dichloroacetyl)amino]-1-(4-nitrophenyl)
propane-Lj-diyi bishexadecanoate (chloramphenicol B. Dissolve about 10 mg in 1 rnL of ethanol
dipalmitate). (50 percent VII-? R, add 3 mL of a 10 gIL solution of ca1dum
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "'Ell
chloride Rand 50 mg of zinc powder R and heat on a water-
bath for 10 min. Filter the hot solution and allow to cool.
Add 0.1 mL of benzoyl chlaride R and shake for I min.
Add 0.5 mL of ferric chloride solution Rl and 2 mL of
chlorokmn R and shake. The upperlayeris light violet-red or
Chloramphenicol Sodium purple.
Succinate C. Dissolve 50 mg in I mL of pyridine R. Add 0.5 mL of
dilute sodium hydroxide solution R and 1.5 mL of waterR. Heat
(ph. Eur. monograph 0709) in a water-bath for 3 min. A red colour develops. Add 2 mL
of nitric acidR and cool under running water. Add I mL of
0.1 Al silvernitrate. A white precipitate is formed slowly.
D. It gives reaction (a) of sodium (2.3.1).
TESTS
pH (2.2.3)
6.4 to 7.0.
Action and use + 5.0 to + 8.0 (anhydrous substance).
Antibacterial. Dissolve 0.50 g in water R and dilute to 10.0 mL with the
Preparation same solvent.
Chloramphenicol Sodium Succinate Injection Chloramphenicol and chloramphenicol dis odium
PhEll _ disuccinate
DEFINITION Test solution Dissolve 25.0 mg of the substance to be
Mixture in variable proportions of sodium (2R,3R)-2- examined in the mobile phase and dilute to 100.0 mL with
[(dichloroacetyl)arnino]-3-hydroxy- 3-(4-nitrophenyl)propyl the mobile phase.
butanedioate (3 isomer) and of sodium (lR,2R)-2-
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1-530 Chlorcyclizine Hydrochloride 2022
Reference solurion (0) Dissolve 10.0 mg of

chloramphenicol CRS in the mobile phase and dilute to
Chlorcyclizine Hydrochloride
100.0 mL with the mobile phase (solution A). Dilute 5.0 mL (ph. Bur. monograph 1086)
of this solution to 100.0 mL with me mobile phase.
Reference solution (b) Dissolve 10.0 mg of chlorampheniwl
disodium disucdnate CRS in the mobile phase and dilute to
100.0 mL with the mobile phase (solution B). Dilute 5.0 mL
of this solution to 100.0 mL with the mobile phase.
Reference solution (c) Dissolve 25 mg of the substance to be
examined in the mobile phase, add 5 mL of solution A and
5 mL of solution B and dilute to 100 mL with the mobile
phase. 337.3 14362-31-3
Column: Action and use
- size: / = 0.25 01, 0 = 4.6 mm; HistamineHI receptor antagonist; antihistamine.
- stationary phase: octadecylsi!Y/ silica gelfor chroma",graphy R
PhElI _
(5 urn).
Mobile phase 20 gIL solution of phosphoric acid R, DEFINmON
methanol R, water R (5:40:55 VIVIV). Chlorcyclizine hydrochloride contains not less than
Flow rate 1.0 mUmin. 99.0 per cent and not more than the equivalent of
Detection Spectrophotometer at 275 om. 101.0 per cent of (RSJ-I-[(4-chlorophenyl)phenylmethyl)-4-
Injection 20 pL methylpiperazine hydrochloride, calculated with reference to
the dried substance.
System suitabl1ity Reference solution (c):
- the 2 peaks corresponding to those in the chromatograms CHARACTERS
obtained with reference solutions (a) and (b) are clearly A white or ahnost white, crystalline powder, freely soluble in
separated from the peaks corresponding to the 2 principal water and in methylenechloride, soluble in alcohol.
peaks in the chromatogram obtained with the test IDENTIFICATION
solution, if necessary, adjust the methanol content of the First identification: B, D.
mobile phase.
Limits:
A. Dissolve 10.0 mg in a 5 gIL solution of sulfuric add Rand
- ch/orampheniwl: not more than the area of the principal
dilute (0 100.0 mL with the same acid. Dilute 10.0 mL of
the solution to 100.0 mL with a 5 gIL solution of sul/un'c
acid R. Examined between 215 nm and 300 nm (2.2.25), the
- chloramphenicol disodium disuccinate: not more than the
solution shows an absorption maximum at 231 run.
The specific absorbance at the maximwn is 475 to 525,
with reference solution (h) (2.0 per cent).
calculated with reference to the driedsubstance.
Water (2.5.12)
B. Examine by infrared absorption spectrophotometry
Pyrogen. (2.6.8) chlorcydjzine hydrochloride CRS. Examine the substances
If intended for use In the manufacture of parenteral prepared as discs.
preparations without a further appropriate procedure for C. Examine the chromatograms obtained in the test for
removal of pyrogens, it complieswith the test for pyrogens. related substances (see Tests). The principal spot in the
Inject per kilogram of the rabbit's mass 2.5 mL of a solution chromatogram obtained with test solution (b) is similar in
in waterfor infec.u'ons R containing 2 mg of the substance to position and size to the principal spot in the chromatogram
be examined per millilitre. obtained with reference solution (a).
ASSAY D. It gives reaction (a) of chlorides (2.3.1).
same solvent. Dilute 5.0 mL of this solution to 100.0 mL
TESTS
with waterR. Measure the absorbance (2.2.25) at the
absorption maximwn at 276 run. Dissolve 0.5 g in water R and dilute to 10 mL with the same
solvent. The solution is clear (2.2.1) and colourless (2.2.2,
Calculate the content of ClsH15Cl2N2NaOs, taking the Method II).
specific absorbance to be 220.
pH (2.2.3)
STORAGE Dissolve 0.10 g in carbon dioxide-free water R and dilute to
In an airtight container, protected from light. If the substance 10 mL with the same solvent.The pH of the solutionis
is sterile, store in a sterile, airtight, tamper-evident container, 5.0 to 6.0.
protected from light.
Related substances
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhElI
Examine by thin-layer chromatography (2.2.27), using a plate
coated with a suitable silica gel.
solvent.
Test solution (b) Dilute 5 mL of test solution (a) to 100 mL
with methanol R.
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2022 Chlordiazepoxide 1-531
*****
Reference solution (a) Dissolve 10 mg of chlorcydisine
Chlordiazepoxide
** **
same solvent. (Ph. Bur. monograph 0656) ***
Reference solution (b) Dissolve 5 mg of methylpiperazine R
in methanol R and dilute to 50 mL with the same solvent
Reference solution (c) Dilute I mL of test solution (b) to
Reference solution (d) Dissolve 10 mg of hydroxyzine
hydrochloride CRS and 10 mg of chiorcydizine
same solvent.
Apply separately to the plate 10 J-IL of each solution and
develop over a path of 15 em using a mixture of 2 volumes 299.8 58-25-3
of concemrated ammonia R, 13 volwnes of methanol Rand
85 volumes of methylene chloride R. Allow the plate to dry in Action and use
air and expose it to iodine vapour for 10 min. In the Benzodiazepine.
chromatogram obtained with [est solution (a): any spot PIIEII ~
corresponding to methylpiperazine is not more intense than

the spot in the chromatogram obtained with reference DEFINITION
solution (b) (0.5 per cent); any spot, apart from the principal 7-Chloro-N-methyl-5-phenyl-3H-I,4-benzodiazepin-2-amine
spot and any spot corresponding to methylpiperazine, is not 4-oxide.
more intense than the spot in the chromatogram obtained Content
with reference solution (c) (0.2 per cent). The test is not 99.0 per cent to 101.0 per cent (dried substance).
valid unless the chromatogram obtained with reference
solution.(d) shows two clearly separated spots, CHARACTERS
Appearance
Almost white or lightyellow, crystalline powder.
drying in an oven at 130°C. Solubility
Practically insoluble in water, sparingly soluble in ethanol
(96 per cent).
Not more than 0.1 per cent) determined on 1.0 g.
ASSAY
Dissolve 0.200 g in a mixture of I mL of 0.1 M hydrochkJric IDENTIFICATION
acid and 50 mL of methanol R. Carry out a potentiometric Infrared absorption spectrophotometry (2.2.24).
titration (2.2.20), using 0.1 M sodium hydroxide. Read Comparison chlordiazepoxide CRS.
the volume added between the two points of inflexion. If the spectra obtained in the solid state show differences,
1 mL of 0.1 M sodium hydroxide is equivalent to 33.73 mg of dissolve the substance to be examined and the reference
ClsH22C12N2' substance separately in m~hylene chloride R, evaporate to
STORAGE
Store protected from light. TESTS
Related substances
IMPURITIES
Liquid chromatography (2.2.29). Cany out the test protected
from bright light and prepare the solutions immediatelY before use.
the mobile phase.
A. N-methylpiperazine. Reference solution (aJ Dilute 1.0 mL of the test solution to
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEII 100.0 mL with the mobile phase. Dilute 2.0 mL of this
solutionto 10.0 mL with the mobilephase.
Reference solution (1)) Dissolve 5 mg of chlordiazepoxide
impurity A CRS in the mobile phase, add 25.0 mL of the test
solution and dilute to 100.0 mL with the mobile phase.
Dilute 2.0 mL of this solution to 50.0 mL with the mobile
phase.
Reference solutUm (c) Dissolve 4.0 mg of
aminochlorobensophenone R in .the mobile phase and dilute to
Column:
- size: I=. 0.15 m, 0 =. 4.6 nun,
- stationary phase: octadecylsilyl silica gelfor chromawgraphy R
(5 urn).
Mobile phase acewnirri/e R, mater R (50:50 VIV).
FIuw rate 1.0 mUmin.
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1-532 Chlordiazepoxide Hydrochloride 2022

Injection 10 ~L.
Run time 6 times the retention time of chlordiazepoxide.
Relattve retention With reference to chlordiazepoxide
(retention time = about 3.6 min): impurity A = about 0.7;
=
impurity B about 2.3; impurity C about 3.9.=
System suitability Reference solution (b): C. (2-amino-5-ehlorophenyl)phenylmethanone
- resolution: minimum 5.0 between the peaks due to (aminochlorobenzophenone).
impurity A and chlordiazepoxide. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE'"
Limits:
- impuritres A, B: for each impurity, not more than the area
of the principal peak in the chromatogram obtainedwith
- impun"ty C: not more than the area of the principal peak
Chlordiazepoxide Hydrochloride
in the chromatogram obtained with reference solution (e) (ph. Eur. monograph 0474)
(0.2 per cent),
- unspecified impurities: foreach impurity, Dot more than
0.5 times the area of the principal peak in me
(0.10 per cent),
peak in the chromatogram obtained withreference
solution (a) (0.5 per cent),
- disregard limil: 0.25 times the area of the principal peak in
(0.05 per cent). 336.2 438-41-5
Loss on drying (2.2.32) Action and use
Maximum 0.5 per cent, determined on 1.000 g by drying in Benzodiazepine.
an oven at 105 "C.
Preparations
Sulfated ash (2.4. f 4) Chlordiazepoxide Capsules
Maximum 0.1 per cent, determined on 1.0 g. Chlordiazepoxide Tablets
ASSAY PhE", ~ _
Dissolve 0.250 g, with heating if necessary, in 80 mL of
anhydrous acetic acid R. Titrate with 0.1 M perchloric acid DEFINITION
determining the end-point potentiometrically (2.2.2U). 7-Chloro-N-methyl-5-phenyl-3H-I,4-benzodiazepin-2-amine
I mL of O.f M perchlari< acid is equivalent to 29.98 mg 4-oxide hydrochloride.
of CI6H'4C1N,O. Content
STORAGE 99.0 per cent to 101.0 per cent (dried substance).
Protected from light. CHARACTERS
Specified impurilies A, B, C. White or slightly yellow, crystalline powder.
Solubility
Soluble in water, sparingly soluble in ethanol (96 per cent).
IDENIlFICATION
Comparison chlordiazepoxide hydrochloride CRS.
If the spectra obtainedin the solid state show differences,
dissolve 100 mg in 9 mL of water R and add I mL of dilure
A. 7-chloro-5-phenyl-1 ,3-dihydro-2H-I,4-benzodiazepin-2- sodium hydroxide solution R. Extract with 10 mL of methylene
one 4-oxide, chloride R in a separating funnel. Evaporate the organic layer
and dry the residue obtained at 100-105 "C. Proceed in the
~
Y'"'Cl same way with the reference substance. Record new spectra
I N using the residues.
CI A '0 B. Dissolve 50 mg in 5 mL of water R, add I mL of dilure
ammonia Rl, mix) aUow to stand for 5 min and filter. Acidify
I'" the filtrate with dilute nitric acidR. The solutiongives
'" reaction (a) of chlorides (2.3. f).
TESTS
B. 6-chloro-2-(chloromethyl)-4-phenylquinazoline 3-oxide,
than reference solution GY6 (2.2.2, J\llethod If).
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2022 Chlorhexidine Acetate 1-533
Dissolve 2.5 g in water R and dilute to 25 mL with the same I mL of 0.1 At! siloer nitrate is equivalent to 33.62 mg
solvent. of C 16H 15CI,NJO.
Liquid chromatography (2.2.29). Carry out thefollowing Protected from light.
operations protected from bright light and prepare the solutions
immediately before use. IMPURITIES
Specified impuruies A, B, C.
the mobile phase.
Reference sdution (b) Dissolve 5 mg of chlordiazepoxide
impurity A CRS in the mobile phase, add 25.0 mL of the test
solution and dilute to 100.0 mL with the mobile phase.
Dilute 2.0 mL of this solution to 50.0 mL with the mobile A. 7-chloro-5-phenyl-1,3-dihydro- 2H- 1,4-benzodiazepin-2-
phase. one d-oxide,
Reference sdution (c) Dissolve 4.0 mg of
aminochlorobenzophenone R in the mobile phase and dilute (0
Column:
=
- size: / 0.15 m, 0 ;;;; 4.6 mID)
- stationary phase: oclOdecylsi/y1 silica gelfor chromaUJgraphy R
(5 pm).
B. 6-ehloro-2-(chloromethyl)-4-phenylquinazoline 3-oxide,
Mobile phase aceUJnitrile R, waterR (50:50 VIV).
Flow rate 1.0 mllrnin.
~
N ",

CI
'" I 0
Injection 10 pL.
Run time 6 times the retention time of chlordiazepoxide.
ReJau've retention With reference to chlordiazepoxide
I'"
h'
= =
(retention time about 3.6 min): impurity A about 0.7;
impurity B = about 2.3; impurity C = about 3.9. C. (2-amino-5-chiorophenyl)phenylmethanone
System suitability Reference solution (b): (aminochlorobenzophenone).
- resolution: minimum 5.0 between the peaks due to _____ ~ PIIEII
impurity A and chlordiazepoxide.
Limits:
- impurities A, B: for each impurity, not more than the area
Chlorhexidine Acetate
- impurity C: not more than the area of the principal peak (Chlorhexidine DiacelOte, Ph. Eur. monograph 0657)
(0.2 per cent),
- unspecified impurities: for each impurity, not more man r""Y~y~y~)
CI
~ NH NH
(0.10 per cent), CI"Q
I '" NH NH
'
~A~A~
peak in the chromatogram obtained with reference h'
626 56-95-1
(0.05 per cent). Action and use
Loss on drying (2.2.32) Antiseptic.
Maximum 0.5 per cent, determined on 1.000 g by drying in Preparation
vacuo at 60 °C for 4 h. Chlorhexidine Irrigation Solution
PhE" _
ASSAY DEFINITION
Dissolve 0.250 g in 50 mL of water R. Titrate with N',N" -(Hexane-1,6-<1iyl)bis[N'-(4-chlorophenyl)
0.1 1\1 silver nitrate, determining the end-point imidodicarbonimidic diamide] diacetate.
potentiometrically (2.2. 2(J). Content
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1-534 Chlorhexidine Acetate 2022
CHARACTERS (96 per cent) Rj transfer each solutionquantitatively to a

Appearance volumetric flask, dilute to 50.0 mL with water Rand allow to
White or almost white, microcrystalline powder. stand for 30 min.
Solubility Measure the absorbance (2.2.25) of each reference solution
Sparingly soluble in water, soluble in ethanol (96 per cent), at 556 nm and plot a calibration curve,
slightly soluble in glycerol and in propylene glycol. Measure the absorbance (2.2.25) of the test solution at
IDENTIFICATION 556 nm. Determine the concentration of chloroaniline from
First identification: A. the calibration curve.
Second identification: B, G. Related substances
Liquid chromatography (2.2.29). Store the solutions at a
temperature notexceeding 12 °G.
Comparison chlorhexidine diacaote CRS. Ten solution Dissolve 0.140 g of the substance to be
B. Thin-layer chromatography (2.2.27). examined in mobile phase A and dilute to 100.0 mL with
Test solution Dissolve 5 mg of the substance to be examined mobile phase A.
in methanol R and dilute to 10 mL with the same solvent. Reference solution (a) Dissolve 5 mg of chlorhexidine for
Reference solution Dissolve 5 mg of chlorhexidine system su;tobihiy CRS (containing impurities A) B) HJ I) KJ N
diaceuue CRS in methanol R and dilute to 10 mL with the and 0) in 1.0 rnL of mobile phase A.
same solvent. Reference solution (b) Dilute 1.0 rnL of the test solution to
Plare TLC $17.00 gel Fm plate R. 100.0 rnL with mobile phase A.
Mobile phase anhydrous formic acidR, water R, ethanol Column:
(96 per <en!! R, melhykne chloride R (7: 10:40:50 VIVIV/JI). - size: 1= 0.25 m, 0 = 4.6 nun;
Application 5 ~ the volume may be adapted based on the - sta,ionary phase: end-copped o,tadecylsi/yl silica gelfor
type of plate used. chromatography R (5 ~m);
Deudopmenc Over 2/3 of the plate. - temperature: 30 "C.
Drying In air. MobIle phase:
- mobile phase A: mix 20 volumes of a 0.1 per cent VIV
Detection A Examine in ultraviolet light at 254 om.
solution of mjfuoroaceric add R in acetonitrile Rand
Results A The principal spot in the chromatogram obtained 80 volumes of a 0.1 per cent VIV solution of m"jluoroa«tic
with the test solution is similar in position and size to the acid R in waterfor chromatography R;
principal spot in the chromatogram obtained with the - mobile phase B: mix 10 volumes of a 0.1 per cent VIV
reference solution. solution of trijluoroaceu·c acid R in waterfor
Deteaion B Treat with copper sulfate solution R and examine chromatography Rand 90 volumes of a 0.1 per cent VIV
in daylight. solutionof tnjfuoroaceuc acid R in aceumitrile R;
with the test solution is similar in position) colour and size to Time MobUe phase A Mobile phase B
(min) (per cent V/V.) (per cent VII')
the principal spot in the chromatogram obtainedwith the
reference solution. 0-2 100 0
2 - 32 lOO --> 80 0-->20
C. It gives reaction (a) of acetates (2.3.1).
32 - 37 80 20
TESTS 37 - 47 80 --> 70 20 --> 30
Impurity P (chloroanllJne) 47 - 54 70 30
Maximum 500 ppm.
Tesrsolution Dissolve 0.20 gin 25 rnL of waterR with Flow rate 1.0 rnIlmin.
shaking if necessary. Add 1 rnL of hydrochloric acid Rand Detection Spectrophotometer at 254 nrn.
dilute to 30 rnL with waterR. Add rapidly and with thorough
mixing after each addition: 5 rnL of a 103 g/l., solution of
Injection 10 pL.
hydrochloric acid R, 0.35 rnL of sodium minte solution R, 2 mL Idendfiauion of impuruies Use the chromatogram supplied
of a 50 gIL solution of ammonium sulfamate R, 5 mL of a with chlarhexidine for system suitability CRS and the
I g/L solution of naphthykthylenediamine dihydrochkiri<k R and chromatogram obtainedwith reference solution (a) to
1 rnL of ethanol (96 per <en!! R; transfer quantitatively to a identify the peaksdue to impurities A) B) H) IJ KJ Nand O.
volumetric flask, dilute to 50.0 mL with waterR and allow to Relative retention With reference to chlorhexidine (retention
stand for 30 min. time = about 35 min): impurity N = about 0.35;
Reference solutions Prepare reference solutions representing = =
impurity B about 0.36; iropurity A about 0.6;
respectively 50 ppm, 100 ppm, 200 ppm, 500 ppm and impurity H = 0.85; iropurity 0 = about 0.90;
600 ppm of chlaroaniline R (impurity P) in the test sample as = =
impurity I about 0.91; impurity K about 1.4.
follows: dilute 1.0 mL, 2.0 rnL, 4.0 rnL, 10.0 rnL and SYStem suitability Reference solution (a):
12.0 rnL of a solution containing O.oIO g/L of chkiraaniline R - peah-to-vaJley ratio: miniroum 2.0, where Hp = height
(impurity P) in di1ure hydrochloric acidR to 20 rnL with above the baseline of the peakdue to impurity Band
waterR. Then, add 10 mL of waterR. Add rapidly and with H v = height above the baselineof the lowest point of the
thorough mixing after each addition: 5 rnL of a 103 w'L curve separating this peak from the peakdue to
solutionof hyc/rochlon·c acid RJ 0.35 mL of sodium nim·te impurity N.
solution R, 2 mL of a 50 gIL solution of ammonium
sulfamtue R) 5 mL of a 1 gIL solution of
naphthylethylenediamine dihydrochloride R and I rnL of ethanol
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2022 Chlorhexidine Acetate 1-535
Limits:
- sum of impurities [ and 0: not more than 0.4 times the
area of me principal peak in the chromatogram obtained
- impurity K: not more than 0.3 times me area of the
- impun"ties A, H, N: for each impurity, not more chan
0.15 times the area of the principal peak in the B. N-[N-[6-[[N-[N-(4-chlorophenyl)
chromatogram obtained with reference solution (b) carbamimidoyl]carbamimidoyl)amino)helC'jI)
(0.15 per cent); carbamimidoyljurea,
- unspecified impun"ties: for each impurity} not more than
(0.10 per cent);
solution (b) (0.8 per cent); E. N-(4-chlorophenyl)guanidine,
(0.05 per cent).
an oven at 105 "C. F. N-(4-chlorophenyl)urea,
ASSAY
Dissolve 0.140 g in 100 mL of anhydrous acetic acid Rand
titrate with 0.1 M perchlonc acid. Determine the end-point
porentiomelrically (2.2.2.11).
I mL of 0.1 M perchknic acid is equivalent to 15.64 mg
of C,.H3SCl,N,oO•.
G. N'-(6-aminohelC'jl)-N'-(4-ehlorophenyl)
IMPURITIES imidodicarbonimidic diamide,
Specified impurities A, H, I, K, N, 0, P.
Other detettable impurities (thefollowing substances would, if
in the monograph. They arelimiud by the general acceptance
criterion for otherlunspedfied impurities andlor by the general
therefore notnecessary «J identify these impurities for
demonstration of compliance. See olso 5.10. Controlofimpurilies
in substances for pharmaceutical use) B~ E, F, G~ M.
H. N',NI' -[azanediylbis(carbonintidoylazanediylhexane-6,1-
diyll]bis[N'-(4-ehlorophenyl)imidodicarbonimidic
diamide],
I. unknown structure,
A. N'-(4-chlorophenyl)-N'-[6-1(N-
cyanocarbamimidoyl)amino]helC'jI)imidodicarbonimidic
diarnlde,
K. N-(4-chlorophenyl)-N'-[N-[6-[[N-[N-(4-
chlorophenyl)carbamimidoyl]carbamimidoyl]amino]
hexyl)carbamimidoyl)urea,
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1-536 Chlorhexidine Gluconate Solution 2022
~~y~y~>
PhE/I ~ _
V NH NH DEFINITION
N',N" -(Hexane-I,6-<1iyl)bis[N3-(4-chlorophenyl)
c''('1 NH NH imidodicarbonimidic diamide] di-n-gluconate.
~~..-Il-~A~ Content
190 gIL to 210 gIL.
CHARACTERS
M.N'-[6-[[N-[N-(4-chlorophenyl)
Appearance
carbamirnidoyljcarbamirnidoyl]aminojhexylj-N3-
Almost colourless or pale-yellowish liquid.
phenylimidodlcarbonimidic diarnide,
Solubility
Miscible with water, with not more than 3 parts of acetone
and with not more than 5 pam of ethanol (96 per cent).
IDENTIFICATION
Second identijicarion: B~ C, D.
Preparation To 1 mL add 40 mL of water R, cool in iced
N. N'-[6-(carbamirnidoylamino)hexylj-N3-(4-chlorophenyl) water) make alkaline to titan yellow paper R by adding
imidodicarbonimidic diamide, dropwlse, and with stirring, strong sodium hydroxide solution R
and add 1 mL in excess. Filter, wash the precipitate with
water R until the washings are free from alkali and
recrystallise from ethanol (70 percent VII? R. Dry at
100-105 °C. Examine the residue.
Comparison chlorhexidine CRS.
Test solution Dilute 10.0 mL of the preparation to be
examined to 50 mL with water R.
O. N'-[6-[[N-[N-(2-ehlorop\1enyl)
Reference solution Dissolve 25 mg of calcium gluconate CRS
carbamimidoyljcarbamirnidoyljaminojhexylj-N'-(4-
in I mL of warer R.
chlorophenyl)imidodicarbonimidic diarnide,
Plate TI£ silica gelplate R.
CI'('1 Mobl7e phase concentrated ammonia R, ethyl acetate R,
waW R, ethanol (96 per <en!> R (10:10:30:50 VIVIVIV).
~NHz Application 5~.
P. 4-chloroaniline.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE/I
Drying At 100"C for 20 min and allow to cool.
Derecrion Spray with a solution containing 25 gIL of
ammonium molybdate R and 10 gIL of cerium sulfate R in dilute
sulfuric acid R, and heat at 110 "C for about 10 min.
Chlorhexidine Gluconate Solution Results The principal spot in the chromatogram obtained
with the test solution is similar in position, colourand size to
(Chlorhexidine Digluconote Solucion, Ph. Eur. the principal spot in the chromatogram obtained with the
monograph 0658) reference solution.
C. To 1 mL add 40 mL of water R, cool in iced water, make
alkaline to tiranyellow paper R by adding dropwise, and with
:~:.~~H
stirring, strong sodium hydroxide solution R and add I mL in
excess. Filter, wash the precipitate with water R until the
HO washings are free from alkali and recrystallise from ethanol
. H (70 per cent VII? R. Dry at 100-105 "C. The residue melts
OH (2.2.14) at 132 "C to 136 "C.
OH 2
D. To 0.05 mL add 5 mL of a 10 gIL solution of cemmide R,
I mL of scrong sodium hydroxide solution R and I mL of
898 18472-51-0 bromine water R; a deep red colouris produced.
Antiseptic. Relative density (2.2.5)
Preparations 1.06 to 1.07.
Chlorhexidine Gluconate Eye Drops pH (2.2.3)
Chlorhexidine GluconateDental Gel 5.5 to 7.0.
Chlorhexidine Irrigation Solution Dilute 5.0 mL to 100 mL with carbon dioxide-free waW R.
Chlorhexidine Mouthwash
Lidocaine and CWorhexidine Gel
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2022 Chlorhexidine Gluconate Solution 1-537
Impurity P (chloroaniline) Time MobUe phase A Mobile phase B

Maxlmum 500 ppm, calculated with reference to (min) (per cent VIV) (per cent VIJ1
chlorhexidine digluconate solution. 0-2 100 0
2·32 100 ---> 80 0--->20
Test solution Dilute 0.20 g of the preparation to be
32·37 80 20
examined to 30 mL with waterR. Add rapidly and with
37 - 47 80 ---> 70 20 ---> 30
thorough mixing after each addition: 5 mL of a 103 gIL
47·54 70 30
solution of hydrochlO1ic acid R, 0.35 mL of sodium min"1e
so/mum R, 2 mL of a 50 gIL solution of ammonium
sulfamate R, 5 mL of a 1 gIL solution of Flow rate 1.0 mUmin.
naphthylethylenediamine dihydrochloride Rand 1 mL of ethanol Detection Spectrophotometer at 254 run.
(96 per cent) R; transfer quantitatively to a volumetric flask, I njeaion 10 ~L.
dilute to 50.0 mL with water R and allow £0 stand for
30 min.
with chlorhexidine for system suilability GRS and the
Reference solutions Prepare reference solutions representing chromatogram obtained with reference solution (a) [Q
respectively 50 ppm, 100 ppm, 200 ppm, 500 ppm and identify the peaks due to impurities A, B, F, G, H, I, J, K, L,
600 ppm of chloroaniline R (impurity P) in the test sample as Nand O.
follows: dilute 1.0 mL, 2.0 rnl., 4.0 mL, 10.0 mL and
Relative retention With reference to chlorhexidine (retention
12.0 mL of a solution containing 0.010 gIL of chforoaniline R
time =: about 35 min): impurity L =: about 0.23j
(impurity P) in dilute hydrochloric acid R to 20 mL with
waterR. Then, add 10 mL of waterR. Add rapidly and with
= =
impurity Q about 0.24; impurity G about 0.25;
= =
impurity N about 0.35; impurity B about 0.36;
impurity F =: about O.5j impurity A = about 0.6j
solution of hydrochloric add R, 0.35 mL of sodium niuiu
impurity H =: about 0.85; impurity 0 =: about 0.90;
sulfamate R, 5 mL of a 1 gIL solution of
= =
impurity I about 0.91; impurity J about 0.96;
naphthylethy/enediomine dihydrochlon"de Rand 1 mL of ethanol
=
(96 per cent) R; transfer each solution quantitatively to a System suitabl7ity Reference solution (a):
volumetric flask, dilute to 50.0 mL with waterRand allow to - resolution: minimum 3.0 between the peaks due to
stand for 30 min. impurities Land G;
- peak-to-valley ratio: minimum 2.0, where H p ::: height
Measure the absorbance (2.2.25) of each reference solution
at 556 nm and plot a calibration curve.
H IJ =: height above the baseline of the lowest point of the
Measure the absorbance (2.2.25) of the test solution at curve separating this peak from the peak due to
556 run. Determine the concentration of chloroaniline from impurity N.
the calibration curve.
Limits:
Related substances - impurity N: not more than the area of the principal peak
Liquid chromatography (2.2.29). Store rhe solutions at a in the chromatogram obtained with reference solution (b)
temperature not exceeding 12 "C. (1.0 per cent);
Testsolution Dilute 1.0 mL of the preparation to be - impuniyH: not more than 0.5 times the area of the
examined to 100.0 mL with mobile phase A. principal peak in the chromatogram obtained with
Reference solution (0) Dissolve 5 mg of chlorhexidine for reference solution (b) (0.5 per cent);
system suitability CRS (containing impurities A, B, F, G, H, I, - impun'ties A) J, K: for each impurity, nor more than
J, K, 1., Nand 0) in 1.0 mL of mobile phase A. 0.4 times the area of the principal peak in the
(0.4 per cent);
- sum of impun'lies I and 0: not more than 0.4 times the
Golumn: area of the principal peak in the chromatogram obtained
- size: I=: 0.25 m, 0 =: 4.6 mm; with reference solution (b) (0.4 per cent);
- stationary phose: end-capped oClodecy/silyl silica gelfor - impun'ty G: not more than 0.3 times the area of the
chromatography R (5 urn), principal peak in the chromatogram obtained with
- temperature: 30°C. reference solution (b) (0.3 per cent),
J.Wobile phase: - impurities B, F, L, Q: for each impurity, not more than
- mobile phase A: mix 20 volumes of a 0.1 per cent VIV 0.2 times the area of the principaJ peak in the
solution of trifiuoroacetic acid R in acetonitrile R and chromatogram obtained with reference solution (b)
80 volumes of a 0.1 per cent VIV solution of tn'jluoroaatk (0.2 per cent);
acid R in waterR; - unspecified impuniies: for each impurity, not more than
- mobile phase B: mix 10 volumes of a 0.1 per cent VIV 0.1 rimes the area of the principal peak in the
solution of trfiuoroacetic acidR in waterR and 90 volumes chromatogram obtained with reference solution (b)
of a 0.1 per cent VIV solution of lnfiuoroaceuc acidR in (0.10 per cent);
acetonitrile R; - total: not more than 3 times the area of the principal peak
(3.0 per cent);
(0.05 per cent).
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1-538 Chlorhexidine Gluconate Solution 2022
ASSAY
Determine the density (2.2.5) of the preparation to be
examined. Transfer 1.00 g to a 250 rnL beaker and add
50 rnL of anhydrous acetic acidR. Titrate with 0.1 M
perchloric acid, determining the end-point potentiometrically
(2.2.20).
1 rnL of 0.1 M perchloric acid is equivalent to 22.44 mg
of C,.,Hs4CI,NlOOI4.
STORAGE H. Nt,N1/-[azanediylbis(carbonimidoylazanediylhexane-6,1-
Protected from light. diyl)]bis[N'-(4-chiorophenyl)imidodicatbonimidic
IMPURITIES diamide],
Specified impurities A, B, F, G, H, I, J, K, L, N, 0, P, Q. I. unknown structure,
Otherdew<table impurities (thefolloroing substances would, if
OH OH
present at a sufficient level, be deuaed by oneor other of the tests H .,
DI
in the monograph. They aretimiud by thegeneral acceptance N 'Y. N~
II 'I ; I OH
criten"on for otherlulISP«ified impurities and/or by the general Cl "'" NyN OH OH
monograph Subseances for pharmaceuti<al use (2034). It is
therefore notnecessary to identify these impurities for
demonstration 0/complianet. See also 5.10. Control of impun"ties
in subSlance.s for pharmaceutical use) E, M. C1U HN\
I NHNHj
-9 ~J--~J--~
J. 1-(4-chlorophenyl)-5-[6-[[4-[(4-chlorophenyl)amino]-6-
[(I S,2R,3R,4R)-1 ,2,3,4,5-pentahydroxypentyl]-1,3, 5-
triazin-2-yl]amino]hexyl]biguanide,
A. N'-(4-chlorophenyl)-N'-[6-[(N-cyanocarbamimidoyl)
amino]hexyl]imidodicarbonimidic diamide,
K. N-(4-chlorophenyl)-N'-[N-[6-[[N-[N-(4-chlorophenyl)
carbamimidoyl]carbamimidoyl]amino]hexylj
carbamimidoyl]urea,
B. N-[N-[6-[[N-[N-(4-chlorophenyl)carbamimidoyl]
carbamimidoyl]amino]hexyl]carbamimidoyl]urea,
E. N-(4-ehlorophenyl)guanidine,
L (5R,6S)-2-[(4-chlorophenyl)amino]-5-hydroxy-6-[(IR,2R)-
1,2,3-trihydroxypropyl]-5,6-dihydro-4H-l,3-oxazin-4-one,
F. N-(4-chlorophenyl)urea,
M.N1-[6-[[N-[N-(4-chlorophenyl)carbamimidoyl]
carbamimidoyl)amino]hexylj-N'-
phenylimidodicarbonimidic diamide,
G. N 1-(6-aminohexyl)-N'-(4-chlorophenyl)
imidodicarbonimidic diamide,
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2022 Chlorhexidine Hydrochloride 1-539

Comparison chlorhexidine dihydrochliJride CRS.
Testsolution Dissolve 5 mg of the substance to be examined
Reference solution Dissolve 5 mg of chlorhexidine
dihydrodlloride CRS in methanol R and dilute to 10 mL with
N. N'-[6-(carbamimidoylamino)hexyl]-N'-(4-chlorophenyl) the same solvent.
imidodicarbonimidic diamide, Plate TLC silica gelFm plat< R.
l\tIobi/e phase anhydrous formic acidR, waler R, ethanol
(96 per cenV R, methylene chloride R (7: 10:40:50 VIVIVIJI).
Application 5 ~L; the volume may be adapted based on the
type of plate used.
Drying In air"
Detection A Examine in ultraviolet light at 254 nm.
O. N'-[6-[[N-[N-(2-chlorophenyl)carbamimidoyl) Results A The principal spot in the chromatogram obtained
carbamimidoyl) amino]hexyl)-N'-(4-<:hlorophenyl) with the test solution is similar in position and size to the
imidodicarbonimidic diamide, principal spot in the chromatogram obtained with the
reference solution.
Detection B Treat with copper sulfate solution R and examine
in daylight.
P. 4-ehloroaniline, with the test solution is similar in position, colour and size to
Q. unknown structure.
reference solution.
__________________ ~ PhE"
TESTS
Impurity P (chIoroaniline)
Chlorhexidine Hydrochloride Maximum 300 ppm.
Testsolution To 0.20 g add 1 mL of hydrochloric acidR,
(ChliJrhexidine DihydrodlliJride, Ph. Bur. monograph shakefor about 30 s, dilute to 30 mL with water R and shake
0659) until a clear solution is obtained. Add rapidly and with
solution of hydrochloric acid R, 0.35 mL of sodium nimt<
sulfamate R, 5 mL of a 1 gIL solution of
. 2 HCI
naphthylethylenediamine dihydrochloride R and I mL of ethanol
(96 per cent) R; transfer quantitatively to a volumetric flask,
dilute to 50.0 mL with water R and allow to stand for
30 min.
Reference solutions Prepare reference solutions representing
578.4 3697-42-5
respectively 50 ppm, 100 ppm, 200 ppm, 300 ppm and
Action and use 500 ppm of chloromiline R (impurity P) in the test sample as
Antiseptic. follows: dilute 1.0 mL, 2.0 mL, 4.0 mL, 6.0 mL and
10.0 mL of a solution containing 0.010 gIL of chloroaniline R
PhE" _ (impurity P) in dilut< hydrochloric acidR to 20 mL with
wat<r R. Then, add 10 mL of wat<r R. Add rapidly and with
DEFINITION
N',N" -(Hexane-I,6-diyl)bis[N'-(4-chlorophenyl)
solution of hydrochloric acid R, 0.35 mL of sodium nimt<
imidodicarbonimidic diamide] dihydrochloride.
Content sulfamate R, 5 mL of a I gIL solution of
98.0 per cent to 101.0 per cent (dried substance). naphthylethylenediamine dihydrochloride Rand 1 mL of ethanol
CHARACfERS (96 per cent) R; transfer each solution quantitatively to a
Appearance volumetric flask, dilute to 50.0 mL with water R and allow to
White or almost white, crystalline powder. stand for 30 min.
Measure the absorbance (2.2.25) of each reference solution
Solubility
Very slightly soluble in water, slightly soluble in propylene at 556 nm and plot a calibration curve.
glycol, very slightly soluble in ethanol (96 per cent). Measure the absorbance (2.2.25) of the test solution at
556 om. Determine the concentration of chloroaniline from
IDENTIFICATION the calibration curve.
Firsr identification: A, C.
Second idemifiauion: B, C.
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1-540 Chlorhexidine Hydrochloride 2022
Related substances chromatogram obtained with. reference solution (b)

Liquid chromatography (2.2.29). Store the solutions al a (0.15 per cent);
temperature not exceeding 12 DC. - unspecified impurities: for each impurity, not more than
Test solution Dissolve 0.130 g of the substance to be 0"1 times the area of the principal peak in the
examined in mobile phase A and dilute to 100.0 mL with chromatogram obtained with reference solution (b)
mobile phase A. (0.10 per cent);
- total: not more than me area of the principal peak in the
Reference solution (aJ Dissolve 5 mg of chlorhexidine for
system suitability CRS (containing impurities A, B, H, r, K, N
(1.0 per cent);
and 0) in 1.0 mL of mobile phase A.
- disregard Iimir. 0.05 times the area of the principal peak in
Reference solution (b) Dilute 1.0 mL of the test solution to me chromatogram obtained with reference solution (b)
100.0 mL with mobile phase A. (0.05 per cent).
Co/umn:
- size: 1= 0.25 m, 0 = 4.6 nun; Maximum 1.0 per cent, determined on 1.000 g by drying in
-suuionatyphase: end-eapped <xradecylsi/yl silica gelfor an oven at 105 DC.
- temperature: 30°C. Sulfated ash (2.4.14)
Maximum 0"1 per cent, determined on 1.0 g.
Mobile phase:
- mobile phaseA: mix 20 volumes ofa 0.1 per cent VIV ASSAY
solution of mftuoroacetic acidR in acetonitrile R and Dissolve 100.0 mg in 5 mL of anhydrous formic acid Rand
80 volumes of a 0.1 per cent VIV solutionof trifluoroautic add 10 mL of acetic anhydride R. Titrate with 0.1 M perchloric
acidR in water for chromatography R; add, determining the end-point potentiometrically (2.2.20).
- mobile phase B: mix 10 volumes of a 0.1 per cent VIV 1 mL of 0.1 M perchlonc acid is equivalent to 14.46 mg
solution of tnjluoroautic add R in waterfor of C"H"CltN,o·
chromatography Rand 90 volumes of a 0.1 per cent VIV
solution of trijluuroauu·c acid R in acetonitrile R; IMPURITIES
Specified impundes A, H, 1, K, N, 0, P.
Time Mobile phase A Mobile phase B Otherdetectable impuniies (thefollowing subsranees would, if
(min) (per cent Vm (per cent Vm present at a sufficient level, be detected by oneor other of the tests
0-2 100 0 in the monograph. They aro limited by thegeneral acceptance
2 - 32 100 _ 80 0_ 20 cruerion for other/unspecified impurities and/or by the general
32 - 37 80 20 monograph Sub,ranees for pharmaceutical use (2034). It is
37 - 47 80 -+ 70 20 _ 30 therefore not neassary to identify these impurities for
47 ~ 54 70 30 demonstration of compliance. See also 5.10. Control of impun"ties
in substances for pharmaceutical use) B, E, F, G, M"
F/qw rate 1.0 mUmin.
Deteaion Spectrophotometer at 254 nm,
Injection 10 pL.
with chlorhexidine for system suitability CRS and the
identify me peaks due to impurities A, B, H, I, K, Nand O.
Relative retention With reference to chlorhexidine (retention
=
time about 35 min): imputity N about 0.35;= A. N'-(4-chIorophenyl)-N'-[6-[(N-
impurity B = about 0.36; impurity A = about 0.6; cyanocarbamimidoyl)amino]hexyl]imidodicarbonimidic
=
Impurity H about 0.85; impurity 0 =
about 0.90; diamide,
= =
imputity I about 0.91; Impurity K about 1.4.
- peak-to-valley ratio: minimum 2.0, where Hp = height
above the baseline of the peak due to impurity B and
Hf) = height above the baseline of the lowest point of the
curve separating this peak from me peakdue to
imputity N.
Limits:
- impun"ty H: not more than 0"5 times the area of the
principal peak in the chromatogram obtained with B. N-[N-[6-[[N-[N-(4-ehlorophenyl)
reference solution (b) (0.5 per cent); carbamimidoyl]carbamimidoyl]amino]hexyl]
- impurity K: not more than 0.4 timesthe area of the carbamimidoyl]urea,
- sum of impurities 1 and 0: not more than 0.4 times the
with. reference solution (b) (0.4 per cent);
- impurities A, N: for each impurity, not more than
0.15 times the area of the principal peak in the E. N-(4-chIoropheuyl)guanidine,
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2022 Chlorinated Lime 1-541
cx
"" I
~y~y~>
Cln
NH NH
Cl
F. N-(-l-chlorophenyljurea,
I"'" NH NH
-'" N
H
.Jl. HN.Jl. HN
O. N'-[6-[[N-[N-(2-chlorophenyI)
carbamimidoyl]carbamimidoyl]amino)hexyl)-N'-(4-
chlorophenyl)imidodicarbonimidic diarnide,
G. N'-(6-aminohexyl)-N'-(4-chlorophenyl)
imidodicarbonimidic diamide,
P. 4-chloroaniline.
_____ ~ f"""
Chlorinated Lime
Action and use
Disinfectant.
H. N1,NI /-[azanediylbis(carbonimidoylazanediylhexane-6,I- DEFINITION

diyl)] bis[N'-(4-<:hlorophenyl)imidodicarbonimidic Chlorinated Lime contains not less than 30.0% wlw of
diamlde], available chlorine, Cl.
I. unknown structure, CHARACTERISTICS
A dull white powder.
Partly soluble in wafer and in ethanol (96%).
IDENTIFICATION
A. Evolves chlorine copiously on the addition of
2M hydrochloric acid.
B. When shaken with water and filtered, the filtrate yields
reaction C characteristic of calcium salts and reaction A
characteristic of chlorides, Appendix VI.
K. N-(4-chlorophenyl)-N'-[N-[6-[[N-[N-(4- ASSAY
chlorophenyl)carbamimidoyl)carbamimidoyl)amino) Triturate 4 g with successive small quantities of water, dilute
hexyl]carbamimidoyl)urea, to 1000 mL with water and shake thoroughly. Mix 100 mL
of the resulting suspension with a solution containing 3 g of
porassium iodide in 100 mL of water, acidify with 5 mL of
6M acetic acid and titrate the liberated iodine with
O.IM sodium thiosulfate VS. Each mL of O.IM sodium
thiosulfate VS is equivalent to 3.545 mg of available chlorine,
Cl.
STORAGE
On exposure to air Chlorinated Lime becomes moist and
gradually decomposes, carbon dioxide beingabsorbed and
M.N1-[6-[[N-[N-(4-chlorophenyl)
chlorine evolved.
carbamimidoyl)carbamimidoyl]amino]hexyl]-N'-
phenylimidodicarbonimidic diarnide,
N. N 1_ [6-(carbamimidoylamino)hexyl)-N'-(-l-chlorophenyl)
imidodicarhonimidic diamide,
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1-542 Chlormadinone Acetate 2022
Chlormadinone Acetate *** Mobae phase:

** ** - mobile phaseA: waterRj
(ph. Eur. monograph 2702) ***** - mobile phase B: acetonitrile R;

0-8 60 '0
8 - 30 60 -> 5 40 ..... 95
30-33 5 95
o
CI
C"H"CIO, 404.9 302-22-7 Injection 10 ilL of test solution (a) and reference
Action and use ldentificatian of impuniies Use the chromatogram supplied
Progestogen. with chlonnadinone acetate for system suitability CRS and the
PhE" _ chromatogram obtained with reference solution (a) to
identify the peaks due to impurities A, B, E and K; use the
DEFINITION chromatogram obtained with reference solution (c) to identify
6-Chloro-3J20-dioxopregna-4,6-dien-17 -yl acetate. the peak due to impurity G.
Content Relative retention With reference to chlormadinone acetate
97.5 per cent to 102.0 per cent (dried substance). (retention time e about 15 min): impurity K;;;; about 0.75;
CHARACTERS =
impurity G about 0.80; impurity A about 0.95; =
impurity E = about 1.04; impurity B = about 1.2.
Appearance
White or almost white, crystalline powder. System suitability:
- signal-co-noise rauo: minimum 35 for the principal peak in
Solubility
the chromatogram obtained with reference solution (b);
Practically insoluble in water, soluble in acetonitrile, slightly
- peab-to-oalley ratio: minimum 1.6, where H p ;;;; height
above the baselineof the peakdue to impurity E and
IDENTIFICATION Hf) ;;;; heightabove the baseline of the lowest point of the
Infrared absorption spectrophotometry (2.2.24). curve separating this peak from the peakdue [Q
Comparison chlonnadinone acetate CRS. chlonnadinoneacetatein the chromatogram obtained
TESTS
- correction factor: multiply the peak area of impurity K by
-14.0 to -10.0 (dried substance).
1.7;
Dissolve 0.200 g in acetonitrile R and dilute to 10.0 mL with - for impurities E, Band K, use the concentration of
the same solvent. chlormadinone acetate in reference solution (b):
Related substances - for impurities other than E, B and K) use the
Liquid chromatography (2.2.29). concentration of impurity G in reference solution (c).
Test solution (a) Dissolve 20 mg of the substance to be Limits:
examined in mobile phase B and dilute to 10.0 mL with - ;mpun'ty B: maximum 0.2 per cent;
mobile phase B. - impurities A, EJ G, K: for each impurity, maximum
Test solution (b) Dissolve 10.0 mg of the substance to be 0.15 per cent;
examined in mobile phase B and dilute to 20.0 mL with - unspecified impurities: for each impurity, maximum
mobile phase B. 0.10 per cent;
Reference solution (a) Dissolve 4 mg of chlonnadinone acetate - total: maximum 0.5 per cent;
~ reporting threshold: 0.05 per cent.
for system suitability CRS (containing impurities A, B, E
and K) in mobile phase B and dilute to 2.0 mL with mobile Loss on drying (2.2.32)
phase B. Maximum 0.5 per cent, determined on 1.000 g by drying in
Reference solution (b) Dilute 1.0 mL of test solution (a) to an oven at 105 "C,
100.0 mL with mobile phase B. Dilute 1.0 mL of this Sulfated ash (2.4.14)
solution to 10.0 mL with mobile phase B. Maximum 0.1 per cent, determined on 1.0 g.
Reference solUlion (c) Dissolve 5.0 mg of chlonnadinone ASSAY
aatate impurity G CRS in mobile phase B and dilute to Liquid chromatography (2.2.29) as described in the test for
50.0 mL with mobile phase B. Dilute 1.0 mL of the solution related substances with the following modifications.
to 50.0 mL with mobile phase B.
MoMe phase Mobile phase B, mobile phase A (45:55 VII').
Reference solution (d) Dissolve 10.0 mg of chlormadinone Injection Test solution (b) and reference solution (d).
acetate CRS in mobile phase B and dilute to 20.0 mL with
mobile phase B. Run time Twice the retention time of chlormadinone
acetate.
Column:
- size: I;;;; 0.15 m, 0 ;;;; 3.0 mm; Retention time Chlonnadinone acetate e about 12 min.
- stationary phase: end..opped extra-dense bonded octadecylsi!YI Calculate the percentage content of C23H29CI04 taking into
silica gelfor chromatography R (3.5 pm). account the assigned content of chlonnadinone acetate CRS.
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2022 Chlormadinone Acetate 1-543
IMPURITIES
Specified impurities A, B, E, OJ K.
Otherdetectable impumies (thefollowing substances would, if
criterion forother/unspecified impurities and/or by the general
CH,
therefore not necessary to identify these impuniies for F. 6~-methyl-3,2o-dioxopregn-4-en-17-yl acetate,
demonstration of compliance. See also 5.10. Control ofimpurities
in substances for pharmaceutical use) C, D, F, H, I, J, L
o
o G. 3,20-dioxopregn-4-en-17-yl acetate,
A. ec-chtoro-3,20-dioxopregn-4-en-17-yl acetate,
Br
H. 3-melhoxy-20-oxopregna-3,5-dien-17-yl acetate,
o
o
CI CH,
B. 2_bromo_6_chloro-3,20-dioxopregna-l,4,6-trien-17-yl
acetate,
o
CI
I. 6-chloro-3-elhoxy-20-oxopregna-3,5-dien-17 -yl acetate,
CI
C. 6~_cWoro_2~_methyl_3,20_dioxopregn_4_en_17_yl acetate,
o o
CH,
CI
-.OyCH3
"/:"/:~ 0 J. 6-chloro_17_hydroxypregna_4,6-<liene_3,20-dione,
o
CI
D. 6-cWoro-3,20-dioxopregna~1,4)6-trien-17-yl acetate,
K. 3,20-dioxopregna-4,6-dien-17-yl acetate,
Br
E. 6_bromo_3,20_dioxopregna-4,6-dien-17-yl acetate,
L. 6!l-cWoro-3,20-dioxopregn-4-en-17-yl acetate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
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1-544 Chlorobutanol 2022
Reference solution (d) Dissolve 50.0 mg of 2-propanol R in

Chlorobutanol the solvent mixture and dilute to 50.0 mL with the solvent
Anhydrous Chlorobutanol mixture. Shake well. Mix 8.0 mL of the solution and 8.0 mL
of reference solution (b), dilute to 200.0 mL with the solvent
(Ph. Bur. monograph 0382) mixture and transfer 5.0 mL to an injection vial.
Colmnn:
- maten'al: fused silica;
- size: I = 30 m, 0 = 0.53 nun;
- stationary phase: cyanopropyl(3)phenyl(3)methyl(94)
CJi,CI,O 177.5 57-15-8 polysi1oxane R (film thickness 3 pm).
Conier gas nitrogen R.
Action and use
Disinfectantpreservative. Flow rate 1.7 mllmin.
PhE,; _
Splil ratio 1:3.
Static head-space conditions that may be used:
DEFINITION - eqUl1ibration temperature: 90 "C;
l,l,I-Trichloro-2-methylpropan-2-ol. - equilibration time: 20 min;
Content - pressurisation time: 30 s.
98.0 per cent to 101.0 per cent (anhydrous substance). Temperature:
CHARACTERS
Tim. Temperature
Appearance (min) ("C)
White or almost white, crystalline powder or colourless Column 0-25 40
crystals, sublimes readily. 25 - 37 40 ~ 220
Solubility 37 - 42 220
Slightly soluble in water, very soluble in ethanol Jnjection port 220
(96 per cent), soluble in glycerol (85 per cent). Detector 230
mp
About 95 "C (without previous drying). Detection Harne ionisation.
IDENTIFICATION Injection 1.0 mL of the test solution and reference
A. Infrared absorption spectrophotometry (2.2.24). solutions (a), (c) and (d).
Comparison chlorobutanol CRS. Identification 0/impurities Use the chromatogram obtained
with reference solution (a) to identify the peakdue to
impurity A; use the chromatogram obtained with reference
TESTS solution (c) to identify the peak due to impurity B; use the
Solution S chromatogram obtainedwith reference solution (d) to
Dissolve 5 g in ethanol (96 per ceno R and dilute to 10 mL identify the peak due to 2-propanol.
with the same solvent. Relativeretention With reference to impurity B (retention
Appearance of solution time = about 14 min): 2-propanol =about 1.1;
Solution S is not more opalescent than reference =
suspension II (2.2.1) and not more intensely coloured than System suitabili'y Reference solution (d):
reference solution BY5 (2.2.2, Method II). - resolution: minimum 1.5 between the peaks due to
Acidity impurity Band 2-propanol.
To 4 mL of solution S add 15 mL of ethanol (96 per cenO R Calculation of contents:
and 0.1 mL of bromothymol blue solution Rl. Not more than - for impurity A, use the concentration of impurity A in
1.0 mL of 0.01 M sodium hydroxide is required to change the reference solution (a);
colour of the indicator to blue. - for impurity B, use the concentration of impurity B in
Impurities A and B reference solution (c).
Head-space gas chromatography (2.2.28). Limas:
Solvent mixture waterR, dimethylformamide R (40:60 VIV). - impun'ty B: maximum 0.10 per cent;
Test solution Introduce 2.000 g of the substance to be - impurity A: maximum 60 ppm.
examined into a vial, add 5.0 mL of the solvent mixture and Chlorides (2.4.4)
close the vial immediately. Maximum 300 ppm.
Reference soltuion (a) Dissolve 60.0 mg of chloroform R Dissolve 0.17 g in 5 mL of ethanol (96 per ceno R and dilute
(impurity A) in the solvent mixture and dilute to 50.0 mL to 15 mL with waterR. When preparing the standard,
with the solvent mixture. Shake well. Dilute 4.0 mL of the replace the 5 mL of waterR by 5 mL of ethanol
solution to 200.0 mL with the solventmixture and transfer (96 per cent) R.
5.0 mL to an injection vial. Water (2.5.12)
Reference 'olulion (b) Dissolve 0.500 g of aceume R Maximum 1.0 per cent, determined on 2.00 g.
(impurity B) in the solvent mixture and dilute to 50.0 mL Sulfated ash (2.4.14)
with the solvent mixture. Shakewell. Maximum 0.1 per cent, determined on 1.0 g.
to 200.0 mL with the solvent mixture and transfer 5.0 mL to
an injection vial.
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2022 Chlorobutanol Hemihydrate 1-545
ASSAY TESTS
Dissolve 50.0 mg in 20 mL of ethanol (96 per cenV R in a Solution S
50 mL centrifuge tube. Add 10 mL of diltue sodium hydroxide Dissolve 5 g in ethanol (96 per cenV R and dilute to 10 mL
solution R, cap tightly and heat in a water-bath for 10 min. with the same solvent.
Cool and transfer quantitatively to a titration vessel using Appearance of solution
100 mL of waterR. Add 20 mL of dilute nitric acid Rand Solution S is not more opalescent thanreference
titrate with 0.1 M silvernitrate, detennining the end-point suspension II (2.2.1) and not more intensely coloured than
potentiometrically (2.2.20). reference solution BY, (2.2.2, Method II).
1 mL of 0.1 M sIlver nitrate is equivalent to 5.92 mg Acidity
of C.H,Cl,O. To 4 mL of solution S add IS mL of ethanol (96 per cenV R
STORAGE and 0.1 mL of bromothymol blue solution Rl. Not more than
In an airtight container. 1.0 mL of 0.01 M sodium hydroxide is required to change the
colour of the indicator to blue.
IMPURITIES
Specified impurities A, B. Impurities A and B
Head-space gas cbromatography (2.2.28).
Solvent mixture waterR, dimethylformamide R (40:60 VIV).
Test soluu'on Introduce 2.000 g of the substance to be
examined into a vial, add 5.0 mL of the solvent mixtureand
A. trichloromethane (chlorofonn), dose the vial immediately.
Reference solution (a) Dissolve 60.0 mg of chlorofOlm R
(impurity A) in the solvent mixture and dilute to 50.0 mL
with the solvent mixture. Shake weU. Dilute 4.0 mL of the
solution to 200.0 mL with the solventmixture and transfer
5.0 mL to an injection vial.
Reference solution (b) Dissolve 0.500 g of acetone R
B. propan-2-one (acetone), (impurity B) in the solvent mixture and dilute to 50.0 mL
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIEur with the solvent mixture. Shake well.
to 200.0 mL with the solvent mixture and transfer 5.0 mL to
an injectionvial.
Chlorobutanol Hemihydrate Reference solution (d) Dissolve 50.0 mg of 2.propanol R in
(Ph. Eur. managraph 0383) mixture. Shake weU. Mix 8.0 mL of the solution and 8.0 mL
NOTE: The name Chlorobutanol was /onnerly usedin the United
of reference solution (b), dilute to 200.0 mL with the solvent
mixture and transfer 5.0 mL to an injection vial.
Kingdom
Column:
- size: 1= 30 m, (2) = 0.53 nun;
- stationary phase: cyanopropyl(3)phenyl(3)methyl(94)
186.5 6001-64-5 poIysi/axane R (film thickness 3 urn).
Oanier gas nitrogen R.
Action and use Flow rate 1.7 mllmin.
Disinfectant preservative. Split ratio 1:3.
PIIEur _ _ ~ _ SUllie head-space conditions that may be used:
- equiJibrat;an temperature: 90 "C;
DEFINITION
- equilibration time: 20 min;
1,I,1-Trichloro-2-methylpropan-2-o1 hemihydrate.
- pressurization time: 30 s.
Content Temperature:
98.0 per cent to 101.0 per cent (anbydrous substance).
CHARACTERS Tim. Temperature
Appearance (min) CC)
White or almost white, crystalline powder or colourless Column 0-25 40
crystals, sublimes readily. 25·37 40 ...,. 220
37 - 42 220
SolublIlty
Injection port 220
Slightly soluble in water, verysoluble in ethanol
Detector 2'0
(96 per cent), soluble in glycerol (85 per cent).
mp
About 78 °C (without previous drying).
Injection 1.0 mL of the test solution and reference
IDENTIFICATION solutions (a), (c) and (d).
Identification 0/impuniies Use the chromatogram obtained
Comparison chlorobutanol hemihydrat< CRS. with reference solution (a) to identify the peakdue to
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1-546 Chlorocresol 2022
impurity A; use the chromatogram obtained with reference

Chlorocresol ***
solution (c) to identify the peak due to impurity B; use the *** ***
chromatogram obtained with reference solution (d) to (Ph. Eur. monograph 0384) ***
identify the peak due to 2-propanol.
Relativeretention With reference to impurity B (retention
. time = about 14 min): 2-propanol = about 1.1; ('1("
=
impurity A about 2.0. Q~
System suitability Reference solution (d): C",
impurity Band 2-propanol. C,H,ClO 142.6 59-5M
Calculation of contents:
- for impurity A, use the concentration of impurity A in Action and use
reference solution (a); Antiseptic; antimicrobial preservative.
- for impurity B, use the concentration of impurity B in
PhE" ~ __
Limits: DEFINITION
- impurity B: maximum 0.10 per cent; 4-Chloro-3-methylphenol.
- impurity A: maximum 60 ppm. Content
Chlorides (2.4.4) 98.0 per cent to 101.0 per cent.
Maximum 100 ppm. CHARACTERS
To I mL of solution S add 4 mL of ethanol (96 percenO R Appearance
and dilute (0 15 mL with water R. When preparing the White or almost white, crystalline powder or compacted
standard, replace the 5 mL of water R by 5 mL of ethanol crystalline masses supplied as pellets or colourless or white
(96 per cenO R. crystals,
Water (2.5.12) Solubility
4.5 per cent to 5.5 per cent, determined on 0.300 g. Slightly soluble in water" very soluble in ethanol
Sulfated ash (2.4.14) (96 per cent), freely soluble in fatty oils. It dissolves in
Maximum 0.1 per cent, determined on 1.0 g. solutions of alkali hydroxides.
ASSAY IDENfIFICATION
Dissolve 50.0 mg in 20 mL of ethanol (96 per <enO R in a A. Melting point (2.2.14): 64"C to 67 "C.
50 mL cenrrifuge tube. Add 10 mL of dilute sodium hydroxide B. To 0.1 g add 0.2 mL of benzoyl chloride Rand 0.5 mL of
solution R, cap tightly and heat in a water-bath for 10 min. dilute sodium hydroxide soluu·on R. Shake vigorously until a
Cool and transferquantitatively to a titration vessel using white, crystalline precipitate is formed. Add 5 mL of water R
100 mL of water R. Add 20 mL of dilute nioic acid R and and filter. The precipitate, recrystaUised from 5 mL of
titrate with 0.1 M silver nimue, determining the end-point methanol R and dried at 70 "C, melts (2.2.14) at 85"C to
potentiometrically (2.2.20). 88 "C.
1 mL of 0.1 M silvernitrate is equivalent to 5.92 mg C. To 5 mL of solution S (see Tests) add 0.1 mL of ferric
of C.H,CI,O. chloride solution RI. A bluish colour is produced.
STORAGE TESTS
In an airtightcontainer. Solution S
IMPURITIES To 3.0 g, finely powdered, add 60 mL of carbon dioxide-free
Specified impurities A, B. water R, shake for 2 min and filter.
than reference solutionBY6 (2.2.2, Method If).
Dissolve 1.25 g in ethanol (96 per <enO R and dilute to
A. trichloromethane (chloroform), 25 mL with the same solvent.
Acidity
To 10 mL of solution S add 0.1 mL of methylredsolution R.
The solution is orange or red. Not more man 0.2 mL of
0.01 M sodium hydroxide is required [0 produce a pure yellow
colour.
Related substances
Gas chromatography (2.2.28): use the normalisation
B. propan-2-one (acetone). procedure.
----~--------------- PhElJf Testsolution Dissolve 1.0 g of the substance to be examined
in aCiwne R and dilute to 100 mL with the same solvent.
100.0 mL with acetQ1le R. Dilute 5.0 mL of this solution to
100.0 mL with acetone R.
Column:
- material: glass;
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2022 Chloroquine Phosphate 1-547
- size: 1= 1.80 m, 0 == 3-4 mm; "'E" _

- stationary phase: silanised diatomaceous earth for gas DEFINITION
chromatography R impregnated with 3-5 per cent m/m of
Chloroquine phosphate contains not less than 98.5 per cent
phenyl(50)methy/(50)poIysiloxane R.
and not more than the equivalent of 101.0 per cent of N'-(7-
Carrier gas nitrogen for chromarography R. chloroquinolin-4-yl}-N1,NI-diethylpentane-l,4-diamine his
Flow rate 30 mUmin. (dihydrogen phosphate), calculated with reference to the
Temperature: dried substance.
- column: 125°C; CHARACTERS
- injeetion port: 210 "C; A white or almost white, crystalline powder, hygroscopic,
- detector: 230°C. freely soluble in water, very slightly soluble in alcohol and in
Detection Flame ionisation. methanol.
Run time 3 times the retention time of chlorocresol. It exists in 2 forms, one of which melts at about 195°C and
Retention time Chlorocresol = about 8 min. the other at about 218 "C.
Limits: IDENTIFICATION
- unspecified impurities: for each impurity, maximum First identification: B, D.
0.10 per cent; Second identification: A, CJ D.
- total: maximwn 1 per cent;
A. Dissolve 0.100 g in warer R and dilute to 100.0 mL with
- disregard limit. the area of the principal peak in the
the same solvent. Dilute 1.0 mL of this solution to 100.0 mL
chromatogram obtained with the reference solution
with waterR. Examined between 210 run and 370 run
(0.05 per cent).
(2.2.25), the solution shows absorption maxima at 220 run,
Non-volatile matter 235 nm, 256 nrn, 329 nm and 342 nm. The specific
Maximum 0.1 per cent. absorbances at the maxima are respectively 600 to 660,
Evaporate 2.0 g to dryness on a water-bath and dry the 350 to 390, 300 to 330, 325 to 355 and 360 to 390.
residue at 100-105 °C. The residue weighs not more than B. Examine by infrared absorption spectrophotometry
2 mg. (2.2.24), comparing with the spectrum obtained with the
ASSAY base isolated from chloroquine sulfate CRS. Record the spectra
In a ground-glass-stoppered flask, dissolve 70.0 mg in 30 mL using solutions prepared as follows: dissolve separately 0.1 g
of glacial acetic acidR. Add 25.0 mL of 0.0167 M potassium of the substance to be examined and 80 mg of the reference
bromate, 20 mL of a 15Q gIL solution of potassium bromide R substance in 10 mL of waterR, add 2 mL of dilute sodium
and 10 mL of hydrochloric acid R. Allow to stand protected hydroxide solution R and shake with 2 quantities, each of
from light for 15 min. Add 1 g of poeassium iodide Rand 20 nIL, of methylene chloride R; combine the organic layers,
100 mL of waterR. Titrate with 0.1 M sodium .hiosulfau, wash with water R, dry over anhydrous sodium sulfate R,
shaking vigorously and using 1 mL of starch solution R, added evaporate to dryness and dissolve the residues separately,
towards the end of the titration, as indicator. Carry out a each in 2 mL of methykne chloride R.
blank titration. C. Dissolve 25 mg in 20 mL of water R and add 8 mL of
I rnL of 0.0167 M potassium bromate is equivalent to pioic acid solution RI. The precipitate, washed with water R,
3565 mg of C,H,ClO. with alcohol R and finally with methylene chloride R, melts
(2.2.14) at 206-209 "C.
STORAGE
D. Dissolve 0.1 gin 10 mL of water R, add 2 mL of dilute
sodium hydroxide solution R and shake with 2 quantities, each
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "'E",
of 20 mL, of methylene chloride R. The aqueous layer,
acidified by the addition of nit"·c acid R, gives reaction (b) of
phosphates (2.3.1).
TESTS
Chloroquine Phosphate Solution S
(Ph. Eur. monograph 0544) Dissolve 2.5 g in carbon dioxide-free waterR and dilute to
and enanllomer Solution S is clear (2.2.1) and not more intensely coloured
than reference solution BYs or GY s (2.2.2, Method11).
pH (2.2.3)
Related substances
Examine by thin-layer chromatography (2.2.27), using SIlica
515.9 50-63-5 gel GF254 R as the coating substance.
Action and use examined in waterR and dilute to 10 rnL with the same
Antiprotozoal (malaria).
solvent.
Preparation Reference solution (a) Dilute 1 mL of the test solution to
Chloroquine Phosphate Tablets 100 mL with waterR.
Reference ,oIution (b) Dilute 5 mL of reference solution (a)
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1-548 Chloroquine Sulfate 2022
Apply to the plate 2 tAL of each solution. Develop over a 235 run, 256 run, 329 om and 342 run. The specific
path of 12 em using a mixture of 10 volwnes of absorbances at the maxima are respectively 730 to 810,
diethylamine R, 40 volwnes of cyclohexane Rand 50 volumes 430 to 470, 370 to 410, 400 to 440 and 430 to 470.
of chloroform R. Allow me plate to dry in air. Examine in B. Examine by infrared absorption spectrophotometry
ultraviolet light at 254 om. Any spot in the chromatogram (2.2.24), comparing with the spectrum obtained with the
obtained with the test solution, apart from the principal spot, base isolated from chloroquine sulfate CRS. Record the spectra
is not more intense than the spot in the chromatogram using solutions prepared as follows: dissolve separately 0.1 g
obtained with reference solution (a) (1.0 per cent) and not of the substance to be examined and of the reference
more than one such spot is more intense than the spot in the substance in 10 mL of water R, add 2 mL of dilute sodium
chromatogram obtained with reference solution (b) hydroxide solution R and shake with 2 quantities, each of
(0.5 per cent). 20 mL, of methylene chloride R; combine the organic layers,
Loss on drying (2.2.32) wash with water R, dry over anhydrous sodium sulfate R,
Maximwn 2.0 per cent, determined on 1.000 g by drying in evaporate to dryness and dissolve the residues separately each
an oven at 105 "C. in 2 mL of methylene chloride R.
ASSAY C. Dissolve 25 mg in 20 mL of water R and add 8 mL of
Dissolve 0.200 g In 50 mL of anhydrous acetic acid R. Titrate picn'c acid solution R1. The precipitate, washed with water R,
with 0.1 M perchkmc acid determining the end-point with erhanol (96 per unO R and finally with erher R, melts
potentiometrically (2.2.20). (2.2.14) at 206"C to 209 "C.
I mL of 0.1 M perchlori< acid is equivalent to 25.79 mg of D. It gives reaction (a) of sulfates (2.3.1).
ClsH32CIN30SP2' TESTS
STORAGE Solution S
In an airtight container, protected from light. Dissolve 2.0 g in carbon dioxide-free water R and dilute [Q
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
than reference solution BY j or GY j (2.2.2, Method Il).
Chloroquine Sulfate ***
*** ***
pH (2.2.3)
Chloroquine Sulphate *** Related substances
(Ph. Eur. monograph 0545) Examine by thin-layer chromatography (2.2.27), using silica
gel GF254 R as the coating substance.
and enanliomel
, H,:SO.. , ~o solvent.
Reference solution (a) Dilute 1 mL of the test solution to
100 mL with water R.
Reference solulion (b) Dilute 5 mL of reference solution (a)
C 18H"CIN,O.S,H,O 436.0
Apply separately to the plate 2 ~L of each solution. Develop
Action and use over a path of 12 em using a mixture of 10 volumes of
Antiprotozoal (malaria). diethylamine R, 40 volumes of cydohexane Rand 50 volumes
of methylene chloride R. Allow the plate to dry in air. Examine
Preparation
in ultraviolet light at 254 nm. Any spot in the chromatogram
Chloroquine Sulfate Tablets
obtained with the lest solution, apart from the principal spot,
PhE<I _ is not more intense than the spot in the chromatogram
obtained with reference solution (a) (1.0 per cent) and not
DEFINITION
more than one such spot is more intense than the spot in the
Chloroquine sulfate contains not less than 985 per cent and
not more than the equivalent of 101.0 per cent of N"-(7- (0.5 per cent).
cWoroquinolin-4-yl)-N1,NI-diethylpentane-I,4-diamine
sulfate, calculated with reference to the anhydrous substance. Water (2. 5. lZ)
CHARACTERS
A white or almost white, crystalline powder, freely soluble in
water and in methanol, very slightly soluble in ethanol
(96 per cent). ASSAY
It melts at about 208 °C (instantaneous method). Dissolve 0.400 gin 50 mL of anhydrous acetic acid R. Titrate
with 0.1 M perchlori< acid determining the end-point
IDENTIFICATION potentiometrically (2.2.20).
I mL of 0.1 M perchloric acid is equlvalenr tc 41.8 mg of
Secondidentification: A, C, D. C,aH,.CIN,O.S.
A. Dissolve 0.100 g in water R and dilute to 100.0 mL with
STORAGE
the same solvent. Dilute 1.0 mL of this solution to 100.0 mL
Store in an airtight container, protected from light.
with water R. Examined between 210 nm and 370 nm
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
(2.2.25), the solution shows absorption maxima at 220 run,
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J
2022 Chloroxylenol 1-549
Chloroxylenol Time Temperature Comment

(Minutes)
OH
0-4 70°
~
isothermal
4-5 70°-->210° linear increase
Me~Me 5 -15 210 isothermal
CI
15 -18 210°-->70° linear gradient
CsH.CIO 156.6 88-04-0 18 - 20 70° re-equilibralion
Action and use

Antiseptic. SYSTEM SUITABILITY
Preparation The test is not valid unless, in the chromatogram obtained
Chloroxylenol Solution with solution (2), the resolution between the peaks due to
tetrachloroethylene and the internal standard is at least 1.5.
DEFINITION
LIMITS
Chloroxylenol is 4-chloro-3,5-xylenol. It contains not less
than 98.0% and not more than 103.0% of CsH.CIO. In the chromatogram obtained withsolution (I), the ratio of
anypeak due to tetrachloroethylene to that of the internal
CHARACTERISTICS standard is not greater thanthe corresponding ratio obtained
White or cream crystals or crystalline powder. It is volatile in in the chromatogram obtained with solution (2) (0.4%).
steam.
Related substances
Very slightly soluble in water, freely soluble in ethanol (96%); Carry out the method for gas chromatography,
soluble in ether, in terpenes and in fixed oils. It dissolves in Appendix m B, using the following solutions in cbtorcform.
solutions of the alkali hydroxides.
(1) 2.0% wlv of the substance beingexamined.
IDENTIFICATION (2) 2.0% w/v of the substance being examined and
The infrared absorption spectrum, Appendix II A J is concordant 0.040% w/v of 4-rhloro-o-cresol (internal standard).
with the reference spectrum of chloroxylenol (RS 055).
TESTS
(a) Use a glass column (1.5 m x 4 mm) packed with acid-
Melting point washed diatomaceous support (80 to 100 mesh) coated with
114° to 116°, Appendix V A. 3% wlw of polyethyleoe glycol (Carbowax 20M is suitable),
Tetrachloroethylene (b) Use nitrogen as the carrier gas at 40 mL per minute.
Carry out the method for gas chromatography,
(c) Use isothermal conditions maintained at 160°.
Appendix ill B, using the following solutions. Prepare a
0.2% vtv solution of butanol (internal standard) in methanol (d) Use an inlet temperature of 200°.
(solution A). (e) Use a flameionisation detector at a temperature of 300°.
(I) To 4 g of the substance being examined, add 5 mL of (I) Inject I ~L of each solution.
solution A and dilute to 25 mL with methanol. SYSTEM SUITABILITY
(2) To 5 mL of a 0.2% vlv solution of terrachloroet/ollene in The test is not valid unless, in the chromatogram obtained
methanol, add 5 mL of solution A and dilute to 25 mL with with solution (2), the resolution between the peaks due to
methanol (equivalent to 0.06488% wlv of tetrachloroethylene in chloroxylenol and d-chloro-o-cresol is at least 1.5
methanol).
LIMITS
In the chromatogram obtained with solution (2) the sum of
(a) Use a fused silica capillary column (30 m x 0.53 mm) the areas of any secondary peaks is not greater than the area of
bonded with a I pm film of polyethylene glyrol 20,000 (RH- the peakdue to the internal standard.
Wax is suitable).
ASSAY
(b) Use hydrogen as the carrier gas at 2 mL per minute.
Dissolve 70 mg in 30 mL of glacial acetic acid, add 25 mL of
(c) Use the gradient conditions described below. 0.0167M potassium bromate VS, 20 mL of a 15% w/v solution
(d) Use an inlet temperature of 240°. of porassium bromide and 10 mL of hydrochloric acid, stopper
(e) Use a flameionisation detector at a temperature of 280°. the flask and allow to stand protected from lightfor
(I) Inject 0.5 ~L of each solution. 15 minutes. Add I g of porassimn iodide and 100 mL of warer
and titrate with O.IM sodimn thiosulfate VS, shaking vigorously
(g) Use a split ratio of 1:20.
and using 1 mL of starch solution, added towards the end of
the titration, as indicator. Repeat the operation without the
substance being examined. The difference between me
titrations represents the amount of potassiwn bromate
required. Each mL of O.0167M potassium bromate VS is
equivalent to 3.915 mg of CsH.CIO.
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1-550 Chlorphenamine Maleate 2022
Reference solution (e) Dissolve the contentsof a vial of

Chlorphenamine Maleate chlorphenamine impun·cy A CRS in 2 mL of the test solution.
(ph. Eur. monograph 0386) Sonicatefor 5 min.
Column:
- size: I ;;;; 0.30 m, (2) ;;;; 3.9 mm;
- srationary phase: o<lade<ylsilyl suica gel for chromawgraphy R
(10 urn).
Mobile phase Mix 20 volumes of aceronitrile Rand
80 volumes of a 8.57 gIL solution of ammonium dihydrogen
phosphate R previously adjusted to pH 3.0 with phosphoric
acid R.
390.9 113-92-8
Fluro rate 1.2 mUmin.
Action and use Deuaion Spectrophotometer at 225 om.
Histamine HI receptor antagonist; antihistamine. Injection 20 ~L
Preparations Rim time 3.5 times the retention time of chforphenemine.
Chlorphenamine Injection Relativeretention With reference to chlorphenamine
Chlorphenamine Oral Solution (retention time « about 11 min): maleic acid e about 0.2j
Chlorphenamine Tablets impurity A = about 0.3; impurity B = about 0.4;
impurity C = about 0.9; impurity D = about 3.0.
PIlE" _
System suilllbiJilY Reference solution (c):
DEFINITION - resolution: minimum 1.5 between the peaks due to
(3RS)-3-(4-Chlorophenyl)-N,N-dimethyl-3-(pyridin-2-yl) impurity C and chlorphenamine.
propan-l-amine hydrogen (Z)-butenedioate. Limits:
Content - correction lactim: for the calculation of contents, multiply
98.0 per cent to 101.0 per cent (dried substance). the peak areas of the following impurities by the
corresponding correction factor: impurity A ;;;; 1.5;
CHARACTERS
Appearance
=
impurity B 1.4;
- impuniy A: not more than 0.4 times the area of the
While or almostwhite, crystalline powder. principal peak in the chromatogram obtained with
Solubility reference solution (a) (0.2 per cent);
Freely soluble in water, soluble in ethanol (96 per cent). - impun·ties B, C, D: for each impurity) not more than
IDENTIFICATION 0.2 times the area of the principal peak in the
A. Melting point (2.2.14): 130°C to 135°C.
(0.1 per cent);
B. Infrared absorption spectrophotometry (2.2.24). - unspedfied impurities: for each impurity, not more than
Comparison chlorphenamine maleate CRS. 0.2 times the area of the principal peakin the
C. Optical rotation (see Tests). chromatogram obtained with reference solution (a)
(0.10 per cent);
TESTS
~ total: not more than the area of the principal peakin the
Solution S
(0.5 per cent);
same solvent.
- disregard limit: the area of the principal peakin the
Appearance of solution chromatogram obtained with reference solution (b)
Solution S is clear (2.2.1) and not moreintensely coloured (0.05 per cent); disregard the peak due to maleic aeid.
thanreference solution BY6 (2.2.2, JWetlwd II).
Loss on drying (2.2.3Z)
Optical rotation (2.2.7) Maximum 0.5 per cent, determined on 1.000 g by drying in
-0.10° to + 0.10 determined on solution S.
0
, an oven at 105°C for 4 h.
Related substances Sulfated ash '(2.4.14)
Testsolution Dissolve0.100 g of the substance £0 be ASSAY
the mobile phase.
with 0.1 M perchloTic add, determining the end-point
Reference solution (a) Dilute 0.5 mL of the test solution to potentiometrically (2.2.20).
Reference solUlwn (b) Dilute 1.0 mL of reference solution (a) of C:.oH2,CIN 20•.
STORAGE
Reference solution (c) Dissolve 5 mg of chlorphenamine
impurity C CRS in 5 mL of the test solution and dilute to
50.0 mL with the mobile phase. Dilute 2 mL of this solution IMPURITIES
to 20 mL with the mobile phase. Specified impurities A, B, C, D.
Reference solution (d) Dissolve 5 mg of 2,2'-d,pyridylamine R
(impurity B) in the mobile phase and dilute to 100 mL with
the mobile phase.
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2022 Chlorpromazine Hydrochloride 1-551
B. Complies with the test for identification ofphenothiazines,

Appendix III A, using chlorpromazine hydrochloride BPCRS to
prepare reference solution.
TESTS
Melting point
56° to 58°, Appendix V A.
A. 2-(4-chlorophenyl)-4-(dimethylamino)-2- [2- Related substances
(dimethylamino) ethyl)butanenitrile, Complies with the test for related substances in phenothiazine!,
Appendix III A, using mobile phase A.
Loss on drying
When dried to constant weightoverphosphorns pemoxide at a
pressure not exceeding 0.7 kPa, loses not more than 0.5% of
its weight. Use 1 g.
B. N-(pyridin-2-yl)pyridin-2-amine (2,2'-dipyridylamine), Sulfated ash
Not more than 0.1%, Appendix IXA.
ASSAY
Dissolve 0.8 g in 300 mL of acetone and carry out Method I
for non-aqu~us titration, Appendix VITI A, using 3 mL of a
saturated solution of methyl orange in acetone as indicator.
Each mL of O.lM perchloric acid VS is equivalent to 31.89 mg
C. (3RS)-3-(4-chlorophenyl)-N-methyl-3-(pyridin-2-yl) of C 17H19ClN2S.
propan-l-amine, STORAGE
Chlorpromazine should be protected from light.
Chlorpromazine Hydrochloride
D. (2RS)-2-( 4-chlorophenyl)-4-(dimethylamino)-2-(Pyridin-2-
yljburanenirrile.
____________________ ""f"
. Hel
Chlorpromazine
355.3 69-09-0
~NMe2
OCN
D Action and use
C' Dopamine receptor antagonist; neuroleptic.
~ I I Preparations
~ s "'"
Chlorpromazine Injection
318.9 50-53-3 Chlorpromazine Oral Solution
Chlorpromazine Tablets
Action and use ""fir ~ _
Preparation DEFINITION
Chlorpromazine Suppositories 3-(2-Chloro-IOH-phenothiazin-10-yl)-N,N-dimethylpropan-
I-amine hydrochloride.
DEFINITION Content
Chlorpromazine is [3-(2-chlorophenothiazin-lO-yl)propyl)- 99.0 per cent to 101.0 per cent (dried substance).
dimethylamine. It containsnot less than 99.0% and not more
than 101.0% ofC 17HI9CIN2S, calculated with reference to CHARACTERS
the dried substance. Appearance
CHARACTERISTICS
Solubility
A white or creamy white powder or waxy solid.
Verysoluble in water, freely soluble in ethanol (96 per cent).
Practically insoluble in water; freely soluble in ethanol (96%)
It decomposes on exposure to airand light.
and in ether.
Ir shows polymorphism (5.9).
IDENTIFICATION
A. The infrared absorption spectrum, Appendix IT A, is IDENTIFICATION
concordant with the reference spectrum of chlorpromazine First identification: A, C.
(RS 056). Second identification: B, C.
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1-552 Chlorpromazine Hydrochloride 2022
A. Infrared absorption spectrophotometry (2.2.24). with the mobile phase. Dilute 1.0 mL of the solution to
Preparation 60 WL solutions in methylene chloride R. 100.0 mL with the mobile phase.
Comparison chlorpromazine hydrochloride CRS. Reference solution (d) Dissolve 4 mg of promazine
B. Identification of phenothiazines by thin-layer hydrochloride CRS (impurity C) and 4.0 mg of chlorpromazin
chromatography (2.3.3): use chlorpromazine hydrochloride CRS impurity E CRS in the mobile phase and dilute to 100.0 mL
to prepare the reference solution. with the mobile phase. Dilute 1.0 mL of the solution to
C. Dissolve 20 mg in 2 mL of methanol R. The solution gives
reaction (a) of chlorides (2.3.1). Column:
- size: I = 0.25 m, 0 = 4.0 rom;
TESTS - stationary phase: base-deactivated oetylsilyl silica gelfor
pH (2.2.3) chromatography R (5 urn).
3.5 to 4.5. Cany outthe Wt proteaed from light and use freshly Mobile phase Mix 0.2 volumes of thicdiethylene glycol R,
prepared solutions. 50 volwnes of acetonimle Rand 50 volumes of a
Dissolve 1.0 g in carbon dioxide-free waterR and dilute to 0.5 per cent VIV solution of lrifiuoroautic acid R previously
10 mL with the same solvent. adjusted to pH 5.3 with tetramethylethylenediamine R.
Impurity F Flow rate 1.0 mUmln..
Thin-layer chromatography (2.2.27). Prepare the solutions Detection Spectrophotometer at 254 am.
immediately before use and proua from light.
Injection 10 ~L.
Solvent mixture dkthylamine R, methanol R (5:95 VIV).
Run time 4 times the retention time of chlorpromazine.
Identification of impurities Use the chromatogram obtained
impurity A; use the chromatogram obtainedwith reference
Reference solution (a) Dissolve the contents of a vialof solution (d) to identify the peaks due to impurities C and E;
chlorpromazine impun"ty F CRS in 2.0 mL of the solvent use the chromatogram obtained with reference solution (a) to
mixture. identify the peak due to impurity D.
Reference solution (b) Dilute 300 J.lL of reference solution (a) Relative retention With reference to chlorpromazine
to 10.0 mL with the solvent mixture. (retention time = about 8 min): impurity A = about 0.4;
Reference solution (c) Dissolve 0.10 g of the substance to be impurity B = about 0.5; impurity C = about 0.7;
examined in the solvent mixture, add 1 mL of reference impurity D = about 0.9; impurity E = about 3.4.
solution (a) and dilute to 5·mL with the solvent mixture. System suitability Reference solution (a):
Plate TLC silica gelF2S4 plate R. - resolution: minimum 2.0 between the peaksdue to
Mobile phase acetone R, diethylamine R, cydohexane R impurity D and chlorpromazine.
(10:10:80 VIVIV). Limits:
Applkation 10 J.lL of the test solution and reference - impurilies B, C~ D: for each impurity, not more than
solutions (b) and (c). 0.6 times the area of the principal peak in the
Development Over 314 of the plate. chromatogram obtained with reference solution (b)
(0.3 per cent);
Dry,;,g In air.
- impun'cy A: not more than 1.5 times the area of the
Detection Examinein ultraviolet light at 254 nm. corresponding peak in the chromatogram obtained with
Retardation factors Impurity F = about 0.5; reference solution (c) (0.15 per cent);
chlorpromazine = about 0.6. - impurity E: not more than 1.5 times the area of the
Systemsuirabilicy Reference solution (c): corresponding peak in the chromatogram obtained with
- the chromatogram shows 2 clearly separated spots due to reference solution (d) (0.15 per cent);
impurity F and chlorpromazine. - unspecified impun'cies: for each impurity, not more than
Limit: 0.2 times the area of the principal peak in the
- impurity F: any spot due to impurity F is not more intense chromatogram obtainedwith reference solution (b)
than the spot in the chromatogram obtained with (0.10 per cent);
reference solution (b) (0.15 per cent). - total: maximum 1.0 per cent;
Related substances the chromatogram obtained with reference solution (b)
Liquid chromatography (2.2.29). Prepare the solutions (0.05 per cent).
immediately before use and protect [rom light. '"
Test solution Dissolve 40.0 mg of the substance to be Maximum 0.5 per cent, determined on 1.000 g by drying in
examined in the mobile phase and dilute to 100.0 mL with an oven at 105°C.
the mobile phase.
Reference solution (a) Dissolve 4 mg of chlorpromazine
impurityD CRS in the mobile phase and dilute to 10 mL
with the mobile phase. To 1 mL of the solution add 1 mL of ASSAY
the test solution and dilute to 100 mL with the mobile phase. Dissolve 0.250 g in a mixture of 5.0 mL of 0.1 M hydrochloric
Reference solution (b) Dilute 1.0 mL of the test solution to acid and 50 mL of ethanol (96 per cenV R. Carry out a
20.0 mL with the mobile phase. Dilute 1.0 mL of this potentiometric titration (2.2. 20), using 0.1 frl sodium
solution to 10.0 mL with the mobilephase. hydroxide. Read the volume added between the 2 points of
inflexion.
Reference solution (c) Dissolve 4.0 rng of chlorpromazine
impurity A CRS in the mobile phase and dilute to 100.0 mL
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2022 Chlorprothixene Hydrochloride 1-553
1 mL of 0.1 1\II sodium hydroxide is equivalent to 35.53 mg of

CI1H20ChN2S.
Chlorprothixene Hydrochloride
STORAGE (Ph. Eur. monograph 0815)
IMPURITIES
Specified impurities A, B, C, D, E, F.
CI
352.3 6469-93-8
Action and use

A. 2-cWoro-10-[3-(dimethylamino)propyl]-5k4 -phenothiazin-
5(10H)-one (chlorpromazine sulfoxide), PU" _
DEFINITION
(3Z)-3-(2-CWoro-9H-thioxanthen-9-ylidene)-N,N-
dimethylpropan-I-amine hydrocWoride.
Content
CHARACTERS
Appearance
B. N'-[3-(2-cWoro-l OH-phenothiazin-IO-yl)propyl]-N' ,N',
White or almostwhite, crystalline powder.
N 3 -trimethylpropane-l,3-diamine,
Solubility
Soluble in water and in ethanol (96 per cent), slightly soluble
in methylenechloride.
mp
About 220 "C.
IDENTIFICATION
First identification: A, E.
C. N,N-dimethyl-3-(IOH-phenothiazin-10-yl)propan-l-amine Second identification: BJ C~ DJ E.
(promazine), A. Infrared absorption spectrophotometry (2.2.24).
Preparation Dissolve 25 mg in 1 mL of water R, add 0.1 mL
of dilute sodium hydroxt"tk sdution R and shake with 2 mL of
methylene chloride R; separate the organic layer and wash with
0.5 mL of water R; evaporate the organic layer to dryness
and dry the residue at 40-50 °C; examine me residue as a
disc.
Comparison Repeat the operations using 25 mg of
chlorprothixene hydrochloride CRS.
D. 3-(2-chloro-1 OH-phenothiazin-IO-yl)-N-methylpropan-I-
amine (demethylchlcrpromazine), B. Dissolve 0.2 g in a mixture of 5 mL of dioxan Rand
5 mL of a 1.5 gIL solution of sodium nitrite R. Add 0.8 mL of
nitric add R. After 10 min, add the solution to 20 mL of
('l water R. Filter the precipate formed after 1 h. The filtrate is
yNH
SO " ,I
CI
used immediately for identification test C. Dissolve the
precipitate by warming in about 15 mL of ethanol
(96 per cenlj R and add the solution to 10 mL of water R.
Filter and dry the precipitate at 100-105 "C for 2 h.
The melting point (2.2.14) is 152 "C to 154 "C.
E. 2-chloro-IOH-phenothiazine, C. To 1 mL of the filtrate obtained in identification test B,
add 0.2 mL of a suspension of 50 mg of/ast redBrait R in
I mL of ethanol (96 per cenlj R. Add I mL of 0.5 M alcoholic
potassium hydroxide. A dark red colour is produced. Carry out
a blank test.
D. Dissolve about 20 mg in 2 mL of nitric add R. A'red
colour is produced. Add 5 mL of water R and examine in.
ultraviolet light at 365 nm. The solution shows green
fluorescence.
F. 3-( -l-chloro-IOH-phenothiazin-l O-yl)-N,N- E. Dissolve 20 mg in 2 mL of water R, acidify with dilute
dimethylpropan-l-amine. nitric add R and allow to stand for 5 min. Centrifuge.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
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1-554 Chlorprothixene Hydrochloride 2022
The supernatant gives reaction (a) of chlorides (2.3.1) Sulfated ash (2.4.14)
1
starting from 'add 0.4 mL of silvernitrate solution RJ • Maximum 0.1 per cent, determined on 1.0 g.
TESTS ASSAY
Solution S Dissolve 0.300 g in a mixture of 5.0 mL of 0.01 M
Dissolve 0.25 g in carbon dioxide-free water R and dilute lO hydrochloric acid and 50 mL of ethanol (96 percenO R. Carry
25 mL with the same solvent. out a potentiometric titration (2.2.20)J using 0.1 M sodium
Appearance of solution hydroxide. Read the volume added between the 2 points of
Solution S is clear (2.2.1) and colourless (2.2.2, MethodIf). inflexion.
pH (2.2.3) I mL of 0.1 M sodium hydroxide is equivalent to 35.23 mg of
4.4 to 5.2 for solution S. C,aH'9 CI2 N S.
liquid chromatography (2.2.29). Cany out the testprouaed Protected from light.
from bright /jgh IMPURITIES
Test solution Dissolve 20.0 mg of the substance to be Specified impurities F.
examined in the mobile phase and dilute to 20.0 mL with Otherdete<table impurities (thefollowing substances would, if
the mobilephase. present at a sufficient levd~ be detected by oneor other of the tests
Reference solut;"n (a) Dilute 1.0 mL of the test solution to in the monograph. They are limited by thegeneral acceptance
100.0 mL with the mobile phase. Dilute 1.0 mL of this cntetion for other/unspecified impun"ties andlor by the general
solution to 10.0 mL with the mobile phase. monograph Substances for pharmaceutical use (2034). It is
Reference solution (b) Dissolve the contents of a vial of therefore not necessary to identify these impurities far
chlorprothixene for system suitability CRS (oontaining demonstration of compliance. See also 5.10. Control of impurities
imputities C and F) in 1 mL of the mobile phase. in substances far phannaceutical use) A B~ C~ D) E.
J
Golumn:
- size: 1 == 0.10 m, 0 = 4.0 mm;
- suuionary phase: base-deactivated end-capped octade<y/sily/
silica gelfor chromatography R (3 ~m).
Mobile phase Solution containing 6.0 gIL of potassium
dihydrogen phosphate R, 2.9 gIL of sodium lauri/sulfate Rand
9 gIL of teoabutylammonium bromide R in a mixture of
50 volumes of methanol R 400 volumes of acetonitrile Rand
J
A. (3RS)-2-chloro-9-[3-(dimethylamino)propyl)-9H-
550 volumes of waterfor chromatography R. thioxanrhen-e-ol,
Equilibnu;"n For about 30 min with the mobile phase.
Injection 20 ~L.
Run time Twice the retention time of chlorprothixene.
with chlorprothixene for system ,uirability CRS and the
chromatogram obtained with reference solution (b) to B. N,N-dimethyl-3-(9H-thioxanthen-9-ylidene)propan-l-
identify the peaks due to impurities C and F. amine,
Relative retention With reference to chlorprorhixene
(retention time = about 10 min): impurity C = about 1.25;
imputity F = about 1.33.
chlorprothixene and imputity C.
Limits:
- impurity F: not more than 5 times the area of the principal
peak in the chromatogram obtained with reference c. (3Z)-3-(2-chloro-9H-thioxanthen-9-ylidene)-N-
solution (a) (0.5 per cent); methylpropan-l-amine,
with reference solution (a) (0.10 percent);
- wtal: not more than 8 times the area of the principal peak
in me chromatogram obtained withreference solution (a)
(0.8 per cent);
- disregard l£mir. 0.5 times the area of the principal peak in
(0.05 per cent). D. (3Z)-3-(4-chloro-9H-thioxanthen-9-ylidene)-N,N-
Loss on drying (2.2.32) dimethylpropan-l-amine,
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2022 Chlortalidone 1-555
:%
s ""
0
I
TESTS
Acidity
Dissolve 1.0 g with heating in a mixture of 25 mL of
aatone Rand 25 mL of carbon dioxide-free water R. Cool.
Titrate with 0.1 1\1 sodium hydroxide, determining the
% Cl
end-point potentiometrically (2.2.20). Not more than
0.75 mL of 0.1 M sodium hydroxide is required.
E. 2-chloro-9H-thioxanlhen-9-one,
Related substances
Solvent mixture Mix 2 volumesof a 2 gIL solution of sodium
hydroxide R, 48 volumes of mobile phase Band 50 volumes
of mobilephase A.
Test solution (aJ Dissolve 50.0 mg of the substance to be
F. (3E)-3-(2-chloro-9H-thioxanthen-9-ylidene)-N,N- Testsolution (b) Dilute 10.0 mL of test solution (a) to
dimethylpropan-I-amine ((E)-chlorprothixene). 100.0 mL with the solvent mixture.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E.. Reference solution (a) Dilute 1.0 mL of test solution (a) to
chlorralidone for system suitablli1y CRS (containing impurities B
Chlortalidone and G) in I mL of the solvent mixture. Dilute 400 ~L of the
(Ph. Eur. monograph 0546) solution to 2 mL with the solventmixture.
~N
00..-8D/
••
I
0 HOo~
~
andenanliomer
Reference solution (c) Dissolve 50.0 mg of chlortalidone CRS
mixture. Dilute 10.0 mL of the solution to 100.0 mL with
me solvent mixture.
CI #"..,.Ii Column:
- size: I::::: 0.25 m, '" : : : 4.6 rnm;
- stationory phase: octylsi!yl suica gelfor chromatography R
338.8 77-35-1 (5 ~m);
Action and use
Thiazide-like diuretic. Mobue phase:
- mobile phase A: dissolve 1.32 g of ammonium p1wsphate R
Preparations in about 900 mL of waterfor chromatography R and adjust
Chlortalidone Tablets to pH 5.5 with duute phosphoric acid R; dilute to 1000 mL
Co-tenldone Tablets with waterfor chromatography R;
P1>E.. ~ _
DEFINITION Tim, Mobile phase A MobUe phase B

2-Chloro-5-[( I RS)-I-hydroxy-3-oxo-2,3-dihydro-IH-isoindol- (min) (per cent VJ1I) (per cent VIJr)
I-yl] benzenesulfonamide. 0- 16 65 35
16 - 21 65 ..... 50 35 ..... 50
Content
21 - 35 50 50
35 - 45 50 ..... 65 50 ---> 35
CHARACIERS
Appearance Flow rote 1.4 mUmin.
White or yellowish-white powder.
Detection Spectrophotometer at 220 ron.
Solubility
injection 20 J.!L of test solution (a) and reference
Very slightly soluble in water, soluble in acetone and in
solutions (a) and (h).
methanol, practically insoluble in methylene chloride.
It dissolves in dilute solutions of alkali hydroxides. Identification of impurities Use the chromatogram supplied
with chlorralidone for system mitabili1y CRS and the
IDENTIFICATION identify the peaks due to impurities B and G.
Infrared absorption spectrophotometry (2.2.24;. Relative retention Withreference to chlortalidone
Comparison chlorralidone CRS. (retention time = about 7 min): impurity B ::::: about 0.7;
U the spectra obtained in the solid state show differences, impurity G = about 6.
dissolve the substance to be examined and the reference System suiuzbi/ily Reference solution (b):
substance separately in methanol R, evaporate to dryness and - resolution: minimum 5.0 between the peaks due to
record new spectra using the residues. impurity Band chlortalidone.
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1-556 Chlortalidone 2022
Limits:
- impunOty B: not more than 7 times the area of the
- impun"ty G: not morethan twice the area of the principal
C. ethyl 2-(4-chloro-3-sulfamoylbenzoyl)benzoate,
area of the principal peak in me chromatogram obtained
with reference solution (a) (O.JO per cent);
peak in the chromatogram obtained withreference
D. 2-cWoro-5-[(1 RS)-I-ethoxy-3-oxo-2,3-dihydro-IH-
(0.05 per cent).
isoindoI-l-yI] benzenesulfonami de,
Chlorides (2.4.4)
Maximum 350 ppm.
o ~
'''XiO
0 H 0
Triturate 0.3 g finely, add 30 mL of water R, shake for 5 min s -,
and filter. 15 mL of the filtrate 'complies with the test. ~ '" I ~ =- and enanliomer
Prepare the standard using 10 mL of chloride standard solution
(5 ppm CO R. CI .&' "

Maximum 0.5 per cent, determined on 1.000 g by drying in E. 2-cWoro-5-[( IRS)-3-oxo-2,3-dihydro-IH-isoindol-l-yl]
an oven at 105°C. benzenesulfonamide,

ASSAY
Injection 20 J1L of test solution (b) and reference
F. (1 13 ,7 13 )_24,66-dichlora-I I ,7 1_dihydroxy_12,13, 72,73 _
solution (c).
tetrahydro-l'H,7'H-3'1.6,5'1.6-dithia-4-aza-I,7(1)-
Calculate the percentage content of C1.JIIlCIN2 0 4S taking diisoindola-2,6(l,3)-dibenzaheptaphan-1 3,3,3,5,5, 73 _
into account the assigned content of chlortalidone CRS. hexone,
IMPURITIES
HOO~ 0
Specified impurities B, G.
Otherdeuaable impun'ties (the following subsUlnces would, if CI~... andenantiomer
present at a sufficient level, be detected by oneor ocher of the tests
in the monograph. They aw limited by thegeneral acceptance
c,V t:
monograph SubsUlnces for pharmaceutical use (2034). It is
G. (3RS)-3-(3,4-dichlorophenyl)-3-hydroxy-2,3-dihydro-IH-
isoindol-l-one,
in substances for pharmaceutical use) A, C, D, E, F, H, J.
o C~H
HO,sxi'6""
I
CI
I
.# ~
andenanllomer
A. 2-(4-cWoro-3-suifobenzoyl)benzoic acid, H. 2-cWoro-5-[(1RS)- 3-oxo-l-[(propan-2-yl)oxy]-2,3-

dihydro-IH-isoindol-I-yl] benzenesulfonamide,
0 0yCH'
o
:d0
0 0
~ II
.... 8 ~ CH3
H,N I I
CI ..# ~
B. 2-(4-cWoro-3-sulfamoylbenzoyl)benzoic acid,
I. propan-2-yl 2-(4-cWoro-3-suifamoylbenzoy])benzoate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P1>E"
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2022 Chlortetracycline Hydrochloride 1-557
Chlortetracycline Hydrochloride *** acid R previously adjusted to pH 2 with concentrated

** ** ammonia R.
(Ph. Eur. monograph 0173) ***** Application I ~L.
Development Over3/4 of the plate.
Drying In air.
with the test solutionis similar in position and size (O the
solution (a).
Compound R Molecular fonnula M, B. To about 2 mg add 5 mL of sulfuric acid R. A deep blue
colour develops and becomes bluish-green. Add the solution
Chlcrreeecrcnoe hydrochloride CI CnH2oI.CI1NzOa 515.3
to 2.5 mL of water R. The colour becomes brownish.
Tetracycline hydrochloride H C:2ZH25ClNzOg 480.9
D. Liquid chromatography (2.2.29) as described in the test
Chlortetracyclinehydrochloride 64-72-2
for related substances with the following modification.
Tetracycline hydrochloride 64-75-5
Action and use Results The principal peak in the chromatogram obtained
Tetracyclineantibacterial. with the test solution is similar in retention time and size to
PhE" _
~

DEFINITION TESTS
Mixture of antibiotics, the main component being the pH (2.2.3)
hydrochloride of (4S,4aS,5aS,6S,12aS)-7-chloro-4- 2.3 to 3.3.
(dimethylamino)-3,6, I0, 12,12a-pentahydroxy-6-methyl-l,11- Dissolve 0.1 g in 10 mL of carbon dioxide-free water R,
dioxo-J,4,4a,5,53,6,11,12a-ocmhydrotetracene-2-carooxamide heating slightly.
(chlortetracycline hydrochloride), a substance produced by
the growth of certain strains of Streptomyces oureofacens or Specific optical rotation (2.2.7)
obtained by any other means.
Content
same solvent.
- chlonetracydine hydrochloride (C"H"CI2N20,): minimum
89.5 per cent (anhydrous substance); Absorbance (2.2.25)
- tetracydine hydrochloride (C 22H.,CIN20,): maximum Maximum 0.40 at 460 run.
6.0 per cent (anhydrous substance); Dissolve 0.125 g in waterR and dilute to 25.0 mL with the
- sum of the conrenrs of chlortetracydine hydrochloride and same solvent.
tetracydine hydrochloride: 94.5 per cent to 102.0 per cent Related substances
(anhydrous substance). Liquid chromatography (2.2.29). Prepare the solutions
CHARACTERS immediately before use.
Appearance Testsolution Dissolve 25.0 mg of the substance to be
Yellowpowder. examined in mobile phase B and dilute to 25.0 mL with
Solubility mobile phase B.
Slightly soluble in water and in ethanol (96 per cent). Reference solution (a) Dissolve 25.0 mg of chlortetracycline
It dissolves in solutions of alkali hydroxides and carbonates. hydrochloride CRS in mobile phase B and dilute to 25.0 mL
with mobile; phase B.
IDENTIFICATION
First identification: C~ D. Reference sotudon (b) Dilute 1.0 mL of the test solution to
100.0 mL with mobile phase B.
Second identification: A J B~ c.
to 10.0 mL with mobile phase B.
Reference solution (d) Dissolve 5 mg of chlortetracycline for
in methanol R and dilute {O 10 mL with the same solvent.
system suitability CRS (containing impurities AJ BJ D J EJ GJ
Reference solution (a) Dissolve 5 mg of chlortetracydine H, J, K and L) in mobile phase B and dilute to 5 mL with
hydrochloride CRS in methanolR and dilute to 10 mL with the mobile phase B.
same solvent.
Reference solution (e) Dissolve 25.0 mg of tetracydine
Reference solution (b) Dissolve 5 mg of chlortetracycline hydrochloride CRS in mobile phase B and dilute to 25.0 mL
hydrochloride CRS, 5 mg of demeclocydine hydrochloride Rand with mobile phase B. Dilute 5.0 mL of this solution to
5 mg of doxycycline R in methanol R and dilute {O 10 mL with 100.0 mL with mobile phase B.
the same solvent.
Column:
Plate TLC octadecylsilyl silica gel F254 plateR. - size: I = 0.075 m, 0 ;;;; 4.6 mm;
l"lobile phase Mix 20 volumes of acetonitrile RJ 20 volumes - stationary phase: end-capped octylsilyl silica gelfor
of methanol Rand 60 volumes of a 63 gILsolution of oxalic chromatography with embedded polargroups R (3.5 urn),
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1-558 Chlortetracycline Hydrochloride 2022
- temperature: 45°C. Water (2.5.12)

Mobile phase: Maximum 2.0 per cent, determined on 0.300 g.
- mobile phase A: to 725 mL of wacer for chromatography R Sulfated ash (2.4. If)
add 50 mL of perchtoric acid sdution R, shake and add Maximum 0.5 per cent, determined on 1.0 g.
225 mL of dimethyl sulfoxide R; Bacterial endotoxins (2.6. If)
- mobile phase B: to 250 rnL of waterfor chromatography R Less than 1 ill/mg, if intended for use in the manufacture of
add 50 mL of perchlom acid solution R, shake and add
700 mL of dimethyl sulfoxide R;
Time Mobile phase A MobUe phase B ASSAY
(min) (per cent VIJi) (per cent VIJI) Liquid chromatography (2.2.29) as described in the test for
0-46 100 --io 0 0 ..... 100 related substances with the following modification.
Injection 10 J.l.L of the test solution and reference
Flow race 0.4 mUmin. solutions (a) and (e).
Detection Spectrophotometer at 280 nrn. Calculate the percentage content of C22H24ChN20S using
Injection 20 ilL of the test solution and reference the chromatogram obtained with reference solution (a) and
solutions (h), (c) and (d). taking into account the assigned content of chlortetracycline
hydrochloride CRS. Calculate the percentage content of
ldenufication of impuniies Use the chromatogram supplied
C22H25C1NzOa using the chromatogram obtained with
with chlortetracycline for system SuiUlbility CRS and the
reference solution (e) and taking into account the assigned
content of tetracydine hydrochloride CRS.
identify the peaks due to impurities A, B, D, E, G, H, J, K
and L. STORAGE
Relauve retention With reference to chlortetracycline Protected from light. If the substance is sterile, store in a
(retention time = about 26 min): impurity D = about 0.5; sterile, airtight, tamper-evident container.
tetracycline = about 0.6; impurity E = about 0.7; IMPURITIES
impurity B = about 0.8, impurity A = about 0.86; Specified impun'ties A, B, D, E, G, H, J, K, L.
impurity G = about 0.9; impurity H = about 1.1; Other detectable impurities (thefollowing substances would, if
=
impurity J about 1.4, impurity K about 1.67; = present at a sufficient level, be detected by oneor other 0/the tests
impurity L = about 1.71.
in the monograph. They are limited by thegeneral a«tjJlance
System suitability Reference solution (d): cricerion for otherlunspecified impurities audJor ~ thegeneral
- resolution: minimum 1.5 between the peaks due to monograph Substances for pharmaceuti<al use (2034). It is
tetracycline and impurity E; minimum 1.5 between the therefore not netessary to identify these impuniies for
peaks due to impurities A and G; minimum 1.5 between demonstration of compliance. See also5.10. Control of impun·tie.s
the peaks due to impurities K and L; if necessary, adjust in substances forphannaaucical use) C, F, 1.
the concentration of dimethyl sulfoxide in mobile
phase A.
Limits:
corresponding correction factor: impurity G = 1.4;
= =
impurity J 0.3; impurity K 0.4; impurity L 0.4; =
- impun"ty A: not more than 4 times the area of the
principal peak in the chromatogram obtained with A. (4R,4aS,5aS,6S,12aSJ-7-chloro-4-(dimethylamino)-
reference solution (b) (4.0 per cent); 3,6,10,12, 12a-pentahydroxy-6-methyl-l, Ll-dioxo-
- impurities B, E: for each impurity, not more than the area J,4,4a,5,53,6,11,12a-octahydrotetracene-2-carboxamide
of the principal peak in the chromatogram obtained with (4-epichlortetracycline),
- impurity J: not more than 3 times the area of the principal
- impurities D J 0, H, L: for each impurity, not more than
twice the area of the principal peak in the chromatogram
obtainedwith reference solution (c) (0.2 per cent);
- impurity K: not more than 1.5 times the area of the
principal peak in the chromatogram obtained with B. (4S,4aS,5aS,6S,12aSJ-7-chloro-4-(dimethylamino)-
reference solution (c) (0.15 per cent); 3,6,10,12,12a-pentahydroxy-I,II-dioxo-
- unspecified impurities: for each impurity, not more than the 1,4,4a,5,53,6,II J Iza-octahydroretracene-2-carboxamide
area of the principal peak in the chromatogram obtained (demeclocycline),
- sum of impurities other than A: not more than twice the
whh reference solution (b) (2.0 per cent);
- disregard limit: 0.5 times the areaof the principal peak in
the chromatogram obtainedwith reference solution (c)
(0.05 per cent).
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2022 Chlortetracycline Hydrochloride 1-559
c. (4R,4aS,5aS,6S,12aS)-4-(dimethylamino)-3,6, I0,12, 12a- H. (4S,4aS,5aS,6S,12.S)-2-acetyl-7-chloro-4-

pentahydroxy-l,11-dioxo-l,4,4a,5,58,6,11,12a- (dimethylamino)-3,6,10,12, 12a-pentahydroxy-6-methyl-
octahydrotetracene-2-carboxamide 4a,5a,6,12a-tetrahydrotett8cene-l, 11(4H,5H)-dione
(4-epidemethyltelracycline), (2-acetyl-2-decarboxamidochlonettacycline),
D. (4R,4aS,5aS,6S,12aS)-4-(dimethyl amino)-3,6, I0,12, 12.- I. (4R,4aS, 12aS)-4-(dimethylamino)-3,1O,12,12a-

pentahydroxy-6-methyl-l, l1-dioxo-l,4,48,5,53,6,ll ,12a- tetrahydroxy-6-methyl-l, Il-dioxo-I,4,4a,5, II, 12a-
octahydrotetracene-2-carboxamide (4-epitetracycline), hexahydrotetracene-2-carboxamide
(4-epianhydrotetracycline),
E. (4R,4aS,5aS,6S, 12aS)-7-chloro-4-(dimethylamino)-
3,6,10,12, 12a-pentahydroxy-I, II-dioxo- J. (4S,4aS, 12aS)-4-(dimethylamino)-3,10,12,12a-
1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-earboxamide tetrahydroxy-fi-methyl-I, ll-dioxo-I ,4,4a,5, 11,12a-
(4-epidemethylcWortetracydine), hexahydrotettacene-2-carboxamide (anhydrotetracycline),
F. (4R,4aS,6S,8aS)-6-[(IR)-7-chloro-4-hydroxy-l-methyl-3- K. (4R,4aS,12aS)-7-chlo,o-4-(dimethylamino)-3,10,12,12a-
oxo-I,3-dihydro-2-benzofuran-I-yl]-4-(dimethylamino)- tetrahydroxy-6-methyl-l, ll-dioxo-I,4,4a,5, II, 12a-
3,8a-dihydroxy-l ,8-dioxo-f,4,4a,5,6, 7,8,8a- hexahydrotettacene-2-carboxamide
octahydronaphthalene-2-carboxanaide (4-epiaohydrochlonetracycline),
(4-epiisochlortetracycline),
L. (4S,4aS,12aS)-7-chloro-4-(dimethylamino)-3,10,12,12a-
tetrahydroxy-6-methyl-I,II-dioxo-I,4,4a,5, II,12a-
G. (4S,4aS,6S,8.S)-6-[(IR)-7-chloro-4-hydroxy-l-methyl-3- hexahydroterracene-2-carboxamide
oxo-I ,3-<lihydro-2-benzofuran-l-yl]-4-(dimethylamino)- (anhydrochlonelracycline).
3,8a-dihydroxy-l,8-dioxo-J ,4,4a,5,6,7,8,8a- _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
octahydronaphthalene-2-carboxamide
(isochlortetracycline),
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1-560 Cholesterol 2022
Acidity
Cholesterol Dissolve 1.0 g in 10 mL of ether R, add 10.0 mL of 0.1 AI
(Ph. Eur. monograph 0993) sodium hydroxide and shakefor about I min. Heat gently to
eliminate ether and then boil for 5 min. Cool) add 10 mL of
waterRand 0.1 mL of phenolphthalein solution R as indicator
and titrate with 0.1 M hydrochloric acid until the pink colour
CH, just disappears) stirring me solutionvigorously throughout
H,C
the titration. Carry out a blank titration. The difference
between the volumes of 0.1 M hydrochloric acid required to
H" change the colour of the indicator in the blank and in the test
HO is not more than 0.3 mL.
386.7 57-88-5 Maximum 0.3 per cent, determined on 1.000 g by drying in
vcuuo at 60 °C for 4 h.
Action and use
Excipient.
PhE" _
ASSAY
DEFINITION Gas chromatography (2.2.28).
Cholest-Scen-Sft-ol, Internal standard soluu·qn Dissolve 0.100 g of pregnenolone
Content isobutyrate CRS in heptane R and dilute to 100.0 mL with the
- cholesterol: minimum 95.0 per cent (dried substance); same solvent.
- total sterols: 97.0 per cent to 103.0 per cent (dried Test solution Dissolve 25.0 mg of the substance to be
substance). examined in me internal standard solution and dilute to
CHARACTERS
Appearance Reference solurian Dissolve 25.0 mg of cholesterol CRS in the
White or almost white, crystaUine powder. internal standard solution and dilute to 25.0 mL with the
same solution.
Solubility
Column:
Practically insoluble in water, sparingly soluble in acetone
and in ethanol (96 per cent).
- size: I;;;;; 30 m, 0 ::::: 0.25 mm;
It is sensitive to light. - suuionary phase: mechylpoly<17oxau, R (film thickness
IDENTIFICATION 0.25 um),
A. Melting point (2.2.14): 147 "C to 150 "C. Carrier gas helium for chromatography R.
B. Thin-layer chromatography (2.2.27). Prepare the solurions Flaw rale 2 m1Jmin.
immediately b<fore use. Split ratio 1:25.
Test solution Dissolve 10 mg of the substance to be Temperature:
examined in ethylene chloride R and dilute to 5 mL with the - column: 275 °C;
same solvent. - inJection pon: 285 °C;
Referenasolution Dissolve 10 mg of cholesterol CRS in - detector: 300 "C.
ethylene chlan"de R and dilute to 5 mL with the same solvent. Detection Flame ionisation.
Plate TLC silica gel G plate R. Injection 1.0 J-1L.
Mobile phase ethyl acetate R, toluene R (33:66 VIV). System suitability Reference solution:
Application 20 ~L. - resolution: minimum 10.0 between the peaks due to
Development Immediately, protected from light, over a path pregnenolone isobutyrate and cholesterol;
of 15 em. - symmetry factor. minimwn 0.6 for the peakdue to
Drying In air. cholesterol.
Detection Spray 3 times with antimony trichloride solution Rj Calculate the percentage content of cholesterol taking into
examine within 3-4 min. account the assigned content of cholesterol CRS. Calculate the
percentage content of total sterols by adding together the
Results The principal spot in the chromatogram obtained contents of cholesterol and other substances with a retention
time less man or equal to 1.5 times the retention time of
cholesterol. Disregard the peaks due to the internal standard
reference solution.
and the solvent.
C. Dissolve about 5 mg in 2 mL of methylene chloride R.
Add I mL of acetic anhydride R, om mL of sulfuric acidR STORAGE
and shake. A pink colouris produced which rapidly changes Protected from light.
to red) then to blue and finally to brilliant green. LABELLING
TESTS The label states the sourcematerial for the production of
Solubility In ethanol (96 per cent) cholesterol (for examplebovine brain and spinal cord, wool
In a stoppered flask) dissolve 0.5 g in 50 mL of elhanol fat or chicken eggs).
(96 percent) R at 50 "C. Allow to stand for 2 h. No deposit
or turbidity is formed.
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2022 Cholesterol for Parenteral Use 1-561
IMPURITIES B. Examine the chromatograms obtained in the assay.

CH,
reference solution.
H,C
C. Dissolve about 5 mg in 2 mL of muhylene chloride R.
Add I mL of acetic anhydride Rand 0.01 mL of su/fun'c
acid R and shake. A pink colour is produced which rapidly
changes to red, then to blue and finally to bright green.
A. 5~-cholest-7-en-3~-01 (larhosrerol),
TESTS
Solubility in ethanol (96 per cent)
In a stoppered flask, dissolve 0.5 g in 50 mL of ethanol
CH, (96 per cen!! R at 50 °C. Allow to stand for 2 h. The solution
is clear.
Acldlty
Dissolve 1.0 g in 10 mL of ether R, add 10.0 mL of O. 1 M
sodium hydroxide and shake for about I min. Heat gently to
eliminate the ether and then boil for 5 min. Cool, add
B. cholesta-5,24-dien-3~-01 (desmosterol), 10 mL of water Rand 0.1 mL of phetwlph'halein sdution R as
indicator and titrate with 0.1 wI hydrochloric acid until the
pink colour just disappears, stirring the solution vigorously
CH, throughout the titration. Carry out a blanktitration.
H,C
The difference between the volumes of 0.1 M hydrochloric
acid required to change the colour of the indicator in the
blank titration and in the test is not more than 0.1 ml.,
Peroxide value (2.5.5, Method A)
Maximum 10.
C. 5~-cholesta-7 ,24-dien-3fl-ol.
Other sterols
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
Gas chromatography (2.2.28): use the normalisation
procedure.
Internal standard solution Dissolve 0.100 g of pngnenolone
;"obutyrate CRS in heptane R and dilute to 100.0 mL with the
Cholesterol for Parenteral Use ***
***••**** same solvent.
examined in the internal standard solution and dilute to
Reference solution Dissolve 25.0 mg of cholesterol CRS in the
CH, internal standard solution and dilute to 25.0 mL with the
H,C same solution.
Column:
- size: 1= 30 m, 0 = 0.25 mm;
- stationary phase: melhyipo/ysiioxane R (Iilm thickness
386.7 " 57-88-5 0.25 urn).
Action and use Canier gas helium for chromatography R.
Excipient. Flow rate 2 mUmin.
PhE<r _ Splitra'w "I:25.
Temperature:
DEFINITION - column: 275 "C;
Cholest-5-en-3fl-ol obtained from Wool fa. (0134). - injection port: 285 "C;
Content - detector: 300 °C.
- cholesterol: 99.0 per cent to 101.0 per cent (dried Detection Flame ionisation.
substance). Injection 1.0 ~L.
CHARACTERS Relative retention With reference to cholesterol (retention
Appearance time = about 8.5 min): pregnenolone
White or almost white, crystalline powder. isobutyrate = about 0.8.
Solubility Syuem suitability Reference solution:
Practically insoluble in water, sparingly soluble in acetone - resolution: minimum 10.0 between me peaksdue to
and in ethanol (96 per cent). pregnenolone isobutyrate and cholesterol.
It is sensitive to light.
IDENTIFICATION
A. Melting point (2.2.14): 147 °C to 150 °C.
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1-562 Choline Salicylate Solution 2022
Limits: Limits:
- total of other substances with a retention time less than or equal - impun'cy A: not more than 0.5 times the area of the
to 1.5 rimes the retention time of cholesterol: maximum corresponding peak in the chromatogram obtained with
0.5 per cent, reference solution (c) (0.05 ppm);
- disregard limit: 0.05 per cent; disregard the peak due to the - impurity B: not more than 0.5 times the area of the
internal standard. corresponding peak in the chromatogram obtained with
Benzoyl ureas reference solution (c) (0.05 ppm).
Liquid chromatography (2.2.29). Loss on drying (2.2.32)
Test solution Dissolve 1.0 g of the substance [Q be examined Maximum 0.1 per cent, determined on 1.000 g by drying in
in 200 mL of heptane R using a magnetic stirrer and add vacuo at 60 °C for 4 h.
10 mL of ocetonitrile R. Shake and allow the layers to Sulfated ash (2.4.14)
separate. Isolate the lower layer (acetonitrile) and add 10 mL Maximum 0.1 per cent, determined on 1.0 g.
of cueumjtrik R to the heptane layerand extract again. Microbial contamination
Combine the lower layers and evaporate to dryness by TAMC: acceptance criterion 10' CFU/g (2.6.12).
suitable means. Add 0.5 mL of acetonitrile R then 05 mL of
water R to the residue. Suspend with the aid of ultrasound Bacterial endotoxlns (2.6.14)
for about 5 min. Centrifuge the suspension for 5 mln and Less than 0.1 ill/mg.
use the supernatant. ASSAY
Reference solution (a) Dissolve 10.0 mg of dijlubenzurcm R Gas chromatography (2.2. 21f) as described in the test for
(impurity A) and 10.0 mg of triflumuTOn R (impurity B) in othersterolswith the following modification:
acetonitrile R and dilute to 100.0 mL with the same solvent. Systemsuitability Reference solution:
Dilute 0.1 mL of the solution to 100.0 mL with acetonitrile R. - symmetry factor. minimum 0.6 for the peakdue to
Reference solution (b) Mix 0.5 mL of reference solution (a) cholesterol.
and 0.5 mL of waterR. Calculate the percentage content of C27H 46 0 taking into
Reference solution (e) Dissolve 1.0 g of the substance to be account the assigned content of cholesterol CRS.
examined in 200 mL of heptane R using a magnetic stirrer. STORAGE
Add 0.5 mL of reference solution (a) and 9.5 mL of Protected from light.
acetonitrile R. Shake and allow the layers to separate. Isolate
the lowerlayer (acetonitrile) and add 10 mL of acetonitrile R IMPURITIES
to the heptane layer and extract again. Combine the lower Specified impun'ties A, B.
layers and evaporate to dryness by suitable means.
ct
Add 0.5 mL of a"tenitrile R then 0.5 mL of water R to the
residue. Suspend with the aid of ultrasound for about 5 min.
Centrifuge the suspension for 5 min and use the supernatant. ~~)l~
F 00
O I""
#
Column: U F
- size: /= 0.25 m, 0 = 3 mmj
- stationary phase: base-deactivated end-eapped IXtadecylsilyl A. 1-(4-ehlorophenyl)-3-(2,6-difluorobenzoyl)urea
silica gd fur chromatography R (5 urn): (diflubenzuron),
Mobile phase:
- mobile phase A: a<etenitrile R, waterfur chromategraphy R
(50:50 VIII);

B. 1-(2-ehlorobenzoyl)-3-[(4-trifluoromethoxy)phenyl]urea
(triflumuron).
o
0-20
20 ~ 20.5
100
100 -+ 0 o -+ 100 _____________________ '''''11
20.5 ~ 30 o 100
After elution of the components, a gradient is applied to

prevent a strongdrifting baselinedue to cholesterol during Choline Salicylate Solution
the following run. Action and use
Flow rate 1 mUmin. Salicylate; non-selective cyclo-oxygenase inhibiror; analgesic;
Detection Spectrophotometer at 254 om. anti-inflammatory.
Injection 100 JIL of the test solution and reference Preparations
solutions (b) and (c). Choline Salicylate Ear Drops
Identification of impwities Use the chromatogram obtained Choline Salicylate Oromucosal Gel
impurities A and B. DEFINITION
Choline Salicylate Solution is an aqueous solutionof choline
Retention time Impurity A = about 10 min;
salicylate. It contains not less than 47.5% wlv and not more
impurity B = about 18 min.
than 52.5% wlv of choline salicylate, ClzH19N04. It may
contain a suitable antimicrobial preservative.
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2022 Choline Theophyllinate 1-563
CHARACTERISTICS CHARACTERISTICS
A clear, colourless liquid. A whire, crystalline powder. It melts between 187° and 192°,
IDENTIFICATION Appendix V A.
A. Mix 0.5 mL with 10 mL of methanol, dry with anhydro", Very soluble in water; soluble in ethanol (96%); veryslightly
sodium sulfate, filter and evaporate the methanol to dryness. soluble in ether.
The infrared absorption spectrum of the residue, Appendix Il A, IDENTIFICATION
is concordant with the reference spectrum of choline salicylate A. The light absorption, Appendix II B, in the range 230 to
(RS 059). 350 urn of a 0.002% wlv solution in O.OIM sodium hydroxide
B. Dilute 5 rnLto 25 mL with WQter. The resulting solution exhibits a maximum only at 275 run. The absorbance at
yields the reactions characteristic of salicylates, Appendix VI. 275 urn is about 0.83.
TESTS B. The infrared absorption spectrum, Appendix II A, is
Acidity concordant with the reference spectrum of choline
Dilute 4 mL to 20 mL with water and add 0.1 mL of phenol theophyllinate (RS 060).
red solution. The solution is yellow and not more than 0.4 mL TESTS
of O.lMsodium hydroxide VS is required to change the colour Clarity and colour of solution
of the solution to reddish violet. 50 mL of a 10% wlv solution is clear, Appendix IV A, and
Clarity and colour of soludon not more intensely coloured than reference solution GY4,
Dilute 1 volume of the solution to 5 volumes with water. Appendix IV B, Method I.
The resulting solution is clear, Appendix IV A, and colourless, Related substances
Appendix IV B, Method II. Carry out the method for thin-/ayer chromatography,
Weight per mL Appendix ill A, using the following solutions of the
1.070 to 1.110 g, Appendix V G. substance being examined in ethanol (96%).
Chloride (1) 1.0% w/v of tile substance being examined.
Mix 0.2 mL with 10 mL of water and add carefully, with (2) 0.010% wlv of the substance being examined.
mixing, 0.1 mL of a mixture of 10 volumes of silver nitrate CHROMATOGRAPHIC CONDITIONS
solution and J volume of nitric acid. The resulting solutionis
not more opalescent than a standard prepared by treating (a) Use as the coating silica gel HF'54'
10 mL of a 0.00164% wlv solution of sodium chloride in the (b) Use the mobile phase as described below.
same manner beginning at the words 'add carefully ... ' (c) Apply 5 IJL of each solution.
(0.1%). . (d) Develop the plate to 15 em.
ASSAY (e) Afterremoval of the plate, dry in air, and examineunder
To 1 g add 50 mL of 1,4-dioxan and 5 mL of acetic anhydride ultraviolet light (254 nm).
and carry out Method I for non-aquecus turation, MOBILE PHASE
Appendix VIII A, using 0.25 mL of methylorange-xylene
5 volumes of ethanol (96%) and 95 volumes of chlorofomt.
cyoool FF solution as indicator. Each mL of O.1M perchlorit
acid VS is equivalent to 24.13 mg of C. 2H ••NO•. Use the LIMITS
weight permL to calculate the percentage of C12HI~04' Any secondary spot in the chromatogram obtained with
weight in volume. solution (1) is not more intense than the spot in the
chromatogram obtained with solution (2) (1%).
Loss on drying
"When dried to constant weight at 105°, losesnot more than
Choline Theophyllinate 0.5% of its weight. Use 1 g.
Sulfated ash
Not more than 0.1 %, Appendix IX A.
+~OH ASSAY
Me,N
For choline
Dissolve 0.6 g in 50 mL of water and titrate with O.05M
sulfuric acid VS, using methyl red mixed solution as indicator,
283.3 4499-40-5 until a violet end point is obtained. Each mL of O. 05M
su/juri< acid VS is equivalent to 12.12 mg of choline,
Action and use CSH15N 0 2 ·
Non-selective phosphodiesterase inhibitor (xanthine), For tlwophyUine
treatment of reversible airways obstruction. To the solution obtained in the Assay for choline, add
Preparation 25 mL of O.lM silvernitrate VS and wann on a water bath for
Choline Theophyllinate Tablets 15 minutes. Cool in ice for 30 minutes, filter and wash the
residue with three 10 mL quantities of water. Titrate the
DEFINITION combined filtrate and washings with O.IM sodium hydroxide
Choline Theophyllinate is choline 1,2,3,6-tetrahydro-I,3- VS. Each mL of 0.1M sodium hydroxide VS is equivalent to
dimethyl-2,6-dioxo-7H-purin-7-ide. It containsnot less than 18.02 mg of theophylline, C,HsN 402 •
41.9% and not more than 43.6% of choline, C5HI5N02 , and STORAGE
not less than 61.7% and not more than 65.5% of
Choline Theophyllinate should be protected from light.
theophylline, C 7H9N402, each calculated with reference to
the driedsubstance.
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1-564 Chondroitin Sulfate Sodium 2022
pH (2.2.3)
Chondroitin Sulfate Sodium 5.5 £0 7.5 for solution 81.
Chondroitin Sulphate Sodium Specific optical rotation (2.2.7)
(Ph. Eur. monograph 2064) -20 to -30 (terrestrial origin) or -12 to -19 (matine origin)
(dried substance), determined on solution S1.
OR' Intrinsic viscosity
C0 RO CO 0.01 m'/kg to 0.15 m'lkg.
~
2Na
v;- OH R = S03Na and R''" H Test solUlion (a) Weigh 5.000 g (mOp) of the substance to be
or examined and add about 80 mL of an 11.7 gIL solution of
OH R =H andR' = SO:JNa
sodium chloride R at room temperature. Dissolve by shaking at
o H3C'w-"'NH
room temperature for 30 min. Dilute to 100.0 mL with an
H OH
oII 11.7 gIL solution of sodium chlond« R. Filter through a
membrane filter (nominalpore size 0.45 11m) and discard the
first 10 mL. The concentration of test solution (a) is only
H,O(C"H"NNa,O"S),
indicative and must be adjusted afteran initial measurement
Action and use of the Viscosity of test solution (a).
Acid mucopolysaccharide; treatment of osteoarthritis. Test solution (b) To 15.0 mL of test solution (a) add
5.0 mL of an 11.7 gIL solution of sodium chloride R.
Ph'" _
Testsolutio" (c) To 10.0 mL of test solution (a) add
DEFINITION 10.0 mL of an 11.7 gIL solution of sodium chloride R.
Natural copolymer based mainly on the 2 disaccharides: (4)- Test solution (d) To 5.0 mL of test solution (a) add
(~-D-g1ucopyranosyluronic acid)-(I ~ 3)-[2-(acetylamino)-2- 15.0 mL of an 11.7 gIL solution of sodium chloride R.
deoXY-~-D-galactopyranosyl 4-sulfate]-(1 ~) and [4)-(i!-o-
Determine the flow-time (2.2.9) for an 11.7 gIL solution of
g1ucopyranosyluronic acid)-(I ~3)-[2-(acetylamino)-2-deoxy
sodium chloride R (to) and the flow times for the 4 test
~-o-gaiactopyranosyl6-sulfate)-(l ~), sodium salr.
solutions (th 12, t3 and 4), at 25.00 ± 0.03 °C. Use an
On complete hydrolysis it liberates n-galacrosarnine,
appropriate suspended level viscometer (specifications:
n-glucuronic acid, acetic acid and sulfuric acid. It is obtained
viscometer constant = about 0.005 mm,z/s2, kinematic
from cartilage of horn terrestrial and marine origins.
viscosity range = 1-5 mm2/s, internal diameter of
Depending on the animal species of origin, it shows different
proportions of 4-sulfate 31)d 6-sulfate groups.
= =
tube R 0.53 mm, volume of bulb C 5.6 mL, internal
=
diameter of tube N 2.8-3.2 mm) with a funnel-shaped
Content lowercapillary end. Use the same viscometer for all
95 per cent to 105 per cent (dried substance). measurements; measure all outflow timesin triplicate.
PRODUCTION The test is not valid unless the results do not differ by more
The animals from which chondroitin sulfate sodium is than 0.35 percent from the mean and if the flow time tl is
derived must fulfil the requirements for the health of animals not less chan 1.6 x to and not more than 1.8 x to. If this is
suitable for human consumption. not the case, adjust the concentration of test solution (a) and
repeat the procedure.
CHARACTERS
Calculation of the relau'1Je viscosities
Appearance
White or almost white, hygroscopic powder. Since the densities of the chondroitin sulfate solutions and of
the solvent are almost equal, the relative viscosities 'lei (being
Solubility llrh TJr2, 'lr3 and 'lrl) can be calculated from the ratio of the
Freely soluble in water, practically insoluble in acetone and flow times for the respective solutions t;, (being th 12, t3 and
in ethanol (96 per cent). (4) to the flow time of the solvent to, but taking into account
IDENTIFICATION the kinetic energycorrection factor for the capillary
A. Infrared absorption spectrophotometry (2.2.24). (B = 30 800 s'), as shown below:
Preparation Discs of potassium bromide R. B
Comparison For chondroitin sulfate sodium of terrestrial £i -t?
originuse chondroitin sulfate sodium CRS and for chondroitin
sulfate sodium of marine origineuse chondroi£in sulJate sodium
-----H
to--
(marine) CRS. tJ
B. Solution SI (see Tests) gives reaction (b) of sodium Calculation oj the concentrations
(2.3.1). Calculate the concentration CI (expressed in kglm3 ) of
C. Examine the electropherograms obtained in the test for chondroitin sulfate sodium in test solution (a) using the
related substances. following expression:
Results The principal band in the electropherogram
obtained with the test solution is similar in position to the x 100-h
"lop x - x - - - x 10
principal band in the electropherogram obtained with 100 100
reference solution (a). x percentage content of chondroilio sulfate sodium as determined
in the assay;
TESTS h loss on drying as a percentage.
Solution SI
Dissolve 2.500 g in 50.0 mL of carbon dioxide-free waterR. Calculate the concentration C2 (expressed in kglm3) of
Solution S2 chondroitin sulfatesodium in test solution (b) using the
Dilute 1.0 mL of solution S I to 10.0 mL with water R. following expression:
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2022 Chondroitin Sulfate Sodium 1-565
Cr xO.75 electrophoresis buffer and the agarose gel. Perform the

Calculate the concentration '3 (expressed in kglmJ ) of electrophoresis under the following conditions: 75 mNgel,
chondroitin sulfate sodium in test solution (c) using the resulting in a voltage of 100-150 V (maximum 300-400 V)
following expression: for a gel of about 12 em x 10 ern. Carry out the
electrophoresis for 12 min. Place the gel in a mixture
c, x 0.50 consisting of 10 volumes of anhydrous ethanol Rand
90 volumes of buffer solution A for 2 min. Carry out the
Calculate the concentration C4 (expressed in kglm 3) of electrophoresis for 20 min. Place the gel in a mixture
chondroitin sulfate sodium in test solution Cd) using the consisting of 30 volumes of anhydrous ethanol Rand
following expression: 70 volumes of buffer solution A for 2 min. Carry out the
c, x 0.25 electrophoresis for 20 min. Stain the gel in the staining
solution for 10 min. Destain the gel for 15 min under
Calculation of the intrinsic viscosity running tap water followed by 10-15 min with warer R until
The specific viscosity 'lsi of the test solution (being rt,» 1152) the band in the eleetropherogram obtained with reference
'ls3 and '1s4) is calculated from the relative viscosities 'In solution (c) is visible. Allow the gel to dry.
(being Ilrt> 11r2' rl-a and 'lr,v according to the following System suiklbilily:
expression: - the electropherograrn obtained with reference solution (c)
shows a visible band;
tlri - 1 - the band in the eleetropherogram obtained with reference
The intrinsic viscosity [11], defined as solution (b) is clearly visible and similar in position to the
band in the eleetropherogram obtained with reference
[~I = lim ('!!.)
c.... o c
solution (a).
Remits Any secondary band in the electropherogram
is calculated by linear least-squares regression analysis using obtained with the test solution is not more intense than the
the following equation: band in the eleetropherogram obtained with reference
solution (b) (2 per cent).
~,; ~ c; X kH + I~J Protein (2.5.33, Melhod 2)
c;
Maximum 3.0 per cent (dried substance).
Co concentration of me substance to be examined expressed Test solurion Dilute 1.0 mL of solution Sl to 50.0 mL with
in kglm 3;
kH Huggins' constar-t.
0.1 M sodium hydroxide.
Reference solutions Dissolve about 0.100 g of bovine
Related substances albumin R, accurately weighed, in 0.1 M sodium hydroxide and
Electrophoresis (2.2.31). dilute to 50.0 mL with the same solvent. Carry out all
Buffer solution A (0.1 M barium acetate pH 5.0). Dissolve additional dilutions using 0.1 M sodium hydroxide.
25.54 g of barium a«tate R in 900 mL of water R. Adjust to Chlorides (2.4.4)
pH 5.0 with glacial acetic acidR and dilute to 1000.0 mL Maximum 0.5 per cent.
with waterR. Dilute I mL of solution S2 to 15 mL with water R. Do not
Buffer solution B (I M barium acetate pH 5.0). Dissolve add diluted nitric acid. Prepare the standard using 5 mL of
255.43 g of barium a«tate R in 900 mL of waterR. Adjust to chloride standardsolution (5 ppm Cl) Rand 10 mL of water R.
pH 5.0 with glacial acetic acid R and dilute to 1000.0 mL Loss on drying (2.2.32)
with water R. Maximum 12.0 per cent, determined on J .000 g by drying in
Staining sdution Dissolve 1.0 g of wlu;dine blue R and 2.0 g an oven at 105°C for 4 h.
of sodium chloride R in 1000 mL of 0.01 M hydrochloric acid. Microbial contamination
Filter. TAMC: acceptance criterion 10' CFU/g (2. 6. 12}.
Tess sohuion Prepare a 30 mg/mL solution of the substance TYMC: acceptance criterion 10' CFU/g (2.6. 12}.
[0 be examined in waterR.
Absence of SUlphylococcus aureus (2.6.13).
Reference solution (a) Prepare a 30 mg/ml; solution of
Absence of Pseudomonas aeruginosa (2.6.13).
chondroitin sulfate sodium CRS in water R.
Absence of Escherichia coli (2.6.J:f).
Reference sohuion (b) Dilute 2.0 mL of reference solution (a)
to 100.0 mL with waterR. Absence of Salmonella (2.6.13).
Reference solution (c) l\lix equal volumes of reference Absence of bile-tolerant gram-negative bacteria (2.6.13).
solution (b) and waterR. ASSAY
Procedure Allow the electrophoresis support to cool the plate Test solution (a) Weigh 0.100 g (m,) of the substance to be
to 10°C. Pre-equilibrate the agarosegel for 1 min in buffer examined, dissolve in waterR and dilute to 100.0 mL with
solution A. Remove excess liquid by careful decanting. the same solvent.
Dry the gel for approximately 5 min. Place 400 mL of buffer Test solution (b) Dilute 5.0 mL of test solution (a) to
solution B into each of the containers of the electrophoresis 50.0 mL with waterR.
equipment. Transfer 1 ul., of each solution to the slots of the Reference solution (a) Weigh 0.100 g (mo) of chondroitin
agarose gel. Pipette a few millilitres of a 50 per cent VIV sulfate sodium CRS, previously dried as described in the test
solution of glycerol R onto the cooled plate of the for Joss on drying, dissolve in water R and dilute EO 100.0 mL
electrophoresis equipment and place the gel in the middle of with the same solvent.
the ceramic plate. Place a wick, saturated with buffer
solution B, at the positive and negative sides of the agarose
gel. Ensure that there is good contact between the
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1-566 Chorionic Gonadotrophin 2022
Titrant solueum (a) Weigh 4.000 g of cerylpyridinium chloride CHARACTERS

monohydrate R and dilute to 1000 mL with waterR. Appearance
Titrant solution (b) Weigh 1.000 g of cerylpyn"dinium chloride White or yellowish-white, amorphous powder.
monohydrate R and dilute to 1000 mL with waterR. Solubility
Perform either visual or photometric titration as follows: Soluble in water.
Visual turation Titrate 40.0 mL of reference solution (a) IDENTIFICATION
and 40.0 mL of test solution (a) with titrant solution (a). When administered to immature rats as prescribed in the
The solution becomes turbid. At the end point, the liquid assay, it causes an increase in the mass of the seminal vesicles
appears clear, with an almost-white precipitate in suspension. and of the prostate gland.
The precipitate is more apparent if 0.1 mL of a 1 per cent
solution of methylene blueR is added before starting the TESTS
titration. The precipitated particles are more apparent against Water (2.5.32)
the blue background. Maximum 5.0 per cent.
Photometric titration Titrate 50.0 mL of reference Bacterial endotoxins (2.6.14)
solution (b) and 50.0 mL of test solution (b) with titrant Less than 0.02 IU per International Unit of chorionic
solution (b). To determine the end point, use a suitable gonadotrophin, if intended for use in the manufacture of
autotitrator equipped with a phototrode at a suitable parenteral preparations without a further appropriate
wavelength (none is critical) in the visible range. procedure for the removal of bacterial endoroxlns.
Calculate the percentage content of chondroitin sulfate ASSAY
sodium using the following expression: The potency of chorionic gonadotrophin is estimated by
comparing under given conditions its effect in increasing the
mass of the seminal vesicles (or the prostate gland) of
inunature rats with the same effect of the International
Standard of chorionic gonadotrophin or of a reference
Vo volwne or appropriate titrant solution when titrating the
epproprtate reference solution, in millilitres;
preparation calibrated in International Units.
VI volume or appropriate titrant sorulton when timuing the The International Unit is the activity contained in a stated
appropriate lest solution. in millilitres; amount of the International Standard, which consists of a
It loss on drying or the subsmnce to be examined, as a percentage;
mixture of a freeze-dried extract of chorionic gonadotrophin
Z percentage content or H20(CI~I~a2014S} .. in chondroitin
5luf(lfe 5«1/iw1 eRS. from the urine of pregnant women with lactose.
The equivalence in International Units of the International
STORAGE Standard is stated by the World Health Organization.
In an airtight container, protected from light. Use immature male rats of the same strain, 19-28 days old,
differing in age by not more than 3 days and having body
LABELLING masses such that the difference between the heaviest and the
The label states the origin of the substance (marine or lightest rat is not more than 109. Assign the rats at random
terrestrial). to 6 equal groups of at least 5 animals. If sets of 6 litter
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PflE"
mates are available, assign one litter mate from each set to
each group and mark according to litter.
Choose 3 doses of the reference preparation and 3 doses of
the preparation to be examined such that the smallest dose is
Chorionic Gonadotrophin sufficient to produce a positive response in some of the rats
and the largest dose does not produce a maximal response in
(ph. Bur. monograph 0498) all the rats. Use doses in geometric progression and as an
Action and use initial approximation total doses of 4 IV, 8 IU and 16 IU
Gonadotrophic hormone. may be tried although the dose will depend on the sensitivity
of the animals used, which may vary widely.
Preparation
Chorionic Gonadotrophin Injection Dissolve separately the total quantities of the preparation to
be examined and of the reference preparation corresponding
PflE" _ to the daily doses to be used in sufficient phosphate-albumin
buffered saline pH 7.2 R such that the daily dose is
DEFINITION
administered in a volume of about 0.5 mL. Add a suitable
Dried preparation containing placental g1ycoproteins obtained
antimicrobial preservative such as 4 gIL of phenol or
from the urine of pregnant women. It has luteinising activity.
0.02 gIL of thiomersal. Store the solutions at 5 ± 3 'C.
Potency
Inject subcutaneously into each rat the daily dose allocated to
Minimum 2500 IV/mg. its group, on 4 consecutive days at the same time each day.
PRODUCTION On the 5th day, about 2~ h after the last injection, euthanise
It is prepared by suitable collection and extraction procedures the rats and remove the seminal vesicles. Remove any
followed by purification steps. extraneous fluid and tissue and weigh the vesicles
The method of preparation includes steps that have been immediately. Calculate the results by the usual statistical
shown to remove and/or inactivate extraneous agents. methods (for example) 5.3) using the mass of the vesicles as
In addition, the process is designed to minimise microbial the response (a suitable correction of the organ mass with
contamination. reference to the body mass of the animal from which it was
taken may be applied; an analysis of covariance may be
It is either dried under reduced pressure or freeze-dried.
used).
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2022 Chymotrypsin 1-567
The estimated potency is not less than 80 per cent and not B. Dilute 0.5 mL of solution S to 5 mL with water R.
more than 125 per cent of the stated potency. Add 0.10 mL of a 20 gIL solution of
The confidence limits (P = 0.95) of the estimated potency tDSy/phenyialanykhloromelhane R in ethanol (96 per cent) R.
are not less than 64 per cent and not more than 156 per cent Adjust to pH 7.0 and shake for 2 h. In a depression in a
of the stated potency. white spot-plate, mix 0.05 mL of this solution with 0.2 mL
STORAGE of the substrate solution (see Identification test A). No colour
develops within 3 min of mixing.
In an airtight container, protected from light" at a
temperature of 2 °C to 8°C. If the substance is sterile, store TESTS
in a sterile, airtight, tamper-evident container. Solution S
LABELLING
10.0 mL with the same solvent,
The label Sial'S:
- the number of International Units per container; Appearance of solution
- the potency, in International Units per milligram. Solution S is not more opalescent than reference
____________________ "'E" suspension II (2.2.1).
pH (2.2.3)
Chymotrypsin 18.5 to 22.5, determined at the absorption maximum at
281 run; maximum 8, determined at the absorption
(Ph. Bur. monograph 0476) minimum at 250 nm.
9004-07-3 Dissolve 30.0 mg in 0.001 M hydrochloric acid and dilute to
Action and use Trypsin
Proteolyt~~ enzyme. Substrate solution To 98.5 mg of rosylarg;nine methylester
"'E" _ hydrochloride R, suitable for assaying trypsin, add 5 mL of
tris(hydroxymethyljaminom,ethane buffersolution pH 8.1 Rand
DEFINITION swirl to dissolve. Add 2.5 mL of methyl red mixed solution R
Chymotrypsin is a proteolytic enzyme obtained by the and dilute to 25.0 mL with water R.
activation of chrymotrypsinogen extracted from the pancreas Test sohuion Transfer to a depression in a white spot-plate
of beef (Bos taulUS L.). It has an activity of not less than 5.0 0.0 I mL of tris(hydroxymethyljamitwmethane buffer solution
microkatals per milligram. In solution it has maximal pH 8.1 Rand 0.1 mL of solution S. Add 0.2 mL of the
enzymic activity at about pH 8; the activity is reversibly substrate solution.
inhibited at pH 3, the pH at which it is most stable.
Reference so/uricm At the same time and in the same manner
PRODUCTION as for the test solution, prepare a solution using the
The animals from which chymotrypsin is derived must fulfil substance to be examined to which not more than
the requirements for the health of animals suitable for human 1 per cent mlm of trypsin BRP has been added.
consumption. Furthermore, the tissues used shall not include Start a timer. No colour appears in the test solution within
any specified risk material as defined by any relevant 3-5 min after the addition of the substrate solution. A purple
international or, where appropriate, national legislation. colour is produced in the control solution.
The method of manufacture is validated to demonstrate that Loss on drying (2.2.32)
the product, if tested, would comply wirh. the following test. Not more than 5.0 per cent, determined on 0.100 g by
Histamine (2.6.11J) drying at 60 °C at a pressure not exceeding 0.7 kPa for 2 h.
Not more than 1 J.tg (calculated as histamine base) per 5
ASSAY
microkatals of chymotrypsin activity. Before carrying out the
The activity of chymotrypsin is determined by comparing the
test, heat the solution of the substance to be examined on a
fate at which it hydrolyses acetyltyrosine ethylester R with the
water-bath for 30 min.
rate at which chymotrypsin BRP hydrolyses the same substrate
CHARACTERS under the same conditions.
Appearance Apparatus Use a reaction vessel of about 30 mL capacity
White or almost white, crystalline or amorphous powder, provided with:
hygroscopic if amorphous. - a device that will maintain a temperature of
Solubility 25.0 ± 0.1 °C;
Sparingly soluble in water. - a stirring device, for example a magnetic stirrer;
- a lid with holes for the insertion of electrodes, the tip of a
IDENTIFICATION
burette, a tube for the admission of nitrogen and the
A. Dilute I mL of solution S (see Tests) to 10 mL with
introduction of reagents.
water R. In a depression in a white spot-plate, mix 0.05 mL
of this solution with 0.2 mL of the substrate solution. An automatic or manual titration apparatus may be used.
A purple colour develops. For the latter, the burette is graduated in 0.005 mL and the
pH meter is provided with a wide-range scale and gJass-
Substrate solution To 24.0 mg of aceo"tyros;ne ethylester R
silver-silver chloride or other suitable electrodes.
add 0.2 mL of ethanol (96 per cent) R and swirl to dissolve.
Add 2.0 mL of 0.067 M phosphare buffer solution pH 7.0 R Test solution Dissolve 25.0 mg of the substance to be
and 1 mL of methylredmixed solution R and dilute to examined in 0.001 . .\1 hydrochloric acid and dilute to
10.0 mL with water R. 250.0 mL with the same acid.
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1-568 Ciclesonide 2022
Reference solution Dissolve 25.0 mg of chymotrypsin BRP in Content

0.001 M hydrochlori< acid and dilute to 250.0 rnL with the 98.0 per cent to 102.0 per cent (anhydrous substance).
same acid.
CHARACTERS
Store the solutions at 0-5 °C. Wann 1 mL of each solution Appearance
to about 25°C over 15 min and use 50 j.1L of each solution White Or yellowish-white, crystalline powder.
(corresponding to about 25 nanokatals) for each titration.
Solubility
Carry out the titration in an atmosphere of nitrogen. Transfer
Practically insoluble in water, freely soluble to soluble in
10.0 mL of 0.01 M calcium chloride solution R to the
reaction vessel and, while stirring, add 0.35 rnL of 0.2 M acetone and in anhydrous ethanol.
acetyltyrosJne ethylester R. When the temperature is steady at IDENTIFICATION
25.0 ± 0.1 °C (after about 5 min), adjust to pH 8.0 exactly A. Infrared absorption spectrophorcmerry (2.2.24).
with 0.02 M sodium hydroxide. Add 50 ~L of the test solution Comparison cidesonid» CRS.
(equivalent to about 5 ~g of the substance to be examined)
and start a timer. Maintain at pH 8.0 by the addition of
0.02 M sodium hydroxide, noting the volume added every Results The principal peakin the chromatogram obtained
30 s. Calculate the volume of 0.02 M sodium hydroxide used with the test solutionis similar in retention time and size to
per second between 30 sand 210 s. Carry out a titration in the principal peak in the chromatogram obtained with
the same manner using the reference solution and calculate reference solution (a).
the volwne of 0.02 M sodium hydroxide used per second. TESTS
Calculate the activity in microkatals per milligram using the Related substances
following expression: Liquid chromatography (2.2.29).
m'xV examined in anhydrous ethanol R and dilute to 50.0 mL with
--xA
mxV' the same solvent.
m massof the substance to be examined. in milligrams; Reference solution (a) Dissolve 50.0 mg of ciclesoniik CRS in
m' mass of dtym~in BRP, in milligrams; anhydrous ethanol R and dilute to 50.0 rnL with the same
V volumeof 0.02 M s«Ii/lm hydTOXide used per secondby the test
solution;
solvent.
V' volume of 0.02 M sodium hydroxide used per secondby the Reference solution (b) Dissolve 3 mg of ciclesoniik
reference solution; imptm'ty B CRS, 3 mg of cidesonide impurity C CRS and 5 mg
A activity of chymouypsin BRP. in microkalals pee milligram.
of cidesonide containing impuril)! A CRS in anhydrous ethanol R
and dilute to 10.0 mL with the same solvent.
STORAGE
Reference solution (c) Dissolve 50 mg of the substance to be
In an airtight container at 2 DC (Q 8 DC, protected from light.
examined in anhydrous ethanol R, add 1.0 mL of reference
LABELLING solution (b) and dilute to 50.0 rnL with anhydrous ethanol R.
The label states: Reference solution (d) Dilute 1.0 rnL of the test solution 10
- the quantity of chymotrypsin and the total activity in 100.0 rnL with anhydrous ethanol R. Dilute 1.0 rnL of this
microkatals per container; solution to 10.0 mL with anhydrous ethanol R.
- for the amorphous substance, mat it is hygroscopic. Column:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""E<I
- size: I = 0.25 m, " = 4.6 mm;
- stationary phase: phenylsilyl $Ilia. gelfor chromatography R
(5 um);
- temperamre: 60 °C.
Ciclesonide Mabile phase water R, anhydrous ethanol R (38:62 VIV).
F!= rate 1.0 mUmin.
Injection 20 r.tL of the test solution and reference
solutions (c) and (d).
Run time 2.2 times the retention time of cic1esonide.
impurities A, Band C.
Relauve retention With reference to ciclesonide (retention
impurity C = about 0.9; impurity A = about 1,4,
540.7 /26544-47-6 System suitability Reference solution (c):
Action and use impurity C and ciclesonide.
Glucocorticoid.
""E<I _ - for each impurity, use the concentration of ciclesonide in
DEFINITION
(2'R)-2 '-Cyclohexyl-II ~-hydroxy-3,20-<lioxo-I6~H-[l,3] Limits:
dioxolo[4',5': 16,17]pregna-1 ,4-<1ien-21-yl - impun·ty A: maximum 1.0 per cent;
2-methylpropanoate.
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2022 Ciclopirox 1-569
- impurities B, C: for each impurity, maximum

Ciclopirox ***
0.15 per cent; *** ***
0.10 per cent;
(Ph. Bur. monograph 1407) ***
uq
- UJlaJ of unspecified impurities: maximum 0.2 per cent;
- total: maximum 1.2 per cent; ,H
N 0
Water (2.5.12) 1#
cli,
207.3 29342-05-0
ASSAY
Liquid chromatography (2.2.29) as described in the test for Action and use
related substances with the following modifications. Antifungal.
PhE" _
Run time 1.6 times the .retention time of cic1esonide.
System sUliability Reference solution (a): DEFINITION
- symmetry factor'. maximum 2.2 for the peakdue to 6-Cyclohexyl-I-hydroxy-4-methylpycidin-2(J H)-one.
ciclesonide. Content
Calculate the percentage content of CJ2H.1407 taking into 98.0 per cent to 101.0 per cent (dried substance).
account the assigned content of cidesonidt CRS. CHARACTERS
Specified impurities A, B, C. White or yellowish-white, crystalline powder.
Solubility
Slightly soluble in water, freely soluble in anhydrous ethanol
and in methylene chloride.
IDENTIFICATION
Secondidentification: A, C.
o
Comparison ci<lopirox CRS.
A. (2'S)-2'-cyclohexyl-II P-hydroxy-3,20-dioxo-16PH-[1,3] C. Thin-layer chromatography (2.2.27).
dioxolo[4',5': 16,17]pregna-I,4-dien-21-yl Test sdution Dissolve 20 mg of the substance to be
2-methylpropanoate (S-epimer of ciclesonide), examined in methanol R and dilute to J0 mL with the same
solvent.
Reference solution Dissolve 20 mg of cidopirox CRS in
Piate TLC silica gelFm plate R.
Pretreatment Before use, predevelop with the mobile phase
until the solvent front has migrated to the top of the plate.
o Allow to dry in air for 5 min.
Mobile phase concentrated ammonia R, water R, ethanol
B. (2' R)-2' -cyciohexyi-Ilp,21-dihydroxy-16PH-[1,3]dioxolo (96 perun!> R (10:15:75 VIV/V).
[4',5': 16,17]pregna-I,4-diene-3,20-dione,
Applirotwn . 10 ~L.
H3C)-CH3
o 0-\.0 Detection A Examine in ultraviolet light at 254 nm.
CH, 0 H Results A The principal spot in the chromatogram obtained
"y
~o"O
and epimer at C' with the test solution is similar in position and size to the
reference solution.
o Detection B Spray with a 20 gIL solution of ferric chloride R
in anhydrous ethanol R.
C. (2'R)-2'-[(lRS)-cyclohex-3-enyl]-IIJl-hydroxy-3,20- Results B The principal spot in the chromatogram obtained
dioxo-16PH-[1,3]dioxolo [4',5': 16,17]pregna-1,4-dien-21- with the test solution is similar in position, colour and size to
yl 2-methylpropanoate. the principal spot in me chromatogram obtainedwith the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" reference solution.
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1-570 Ciclopirox 2022
TESTS - iff/pun'ties B, C at 298 nm: for each impurity, not more

Appearance of solution than the area of me peak due to impurity B in the
The solution is clear (2.2.1) and not more intensely coloured chromatogram obtained with reference solution (b)
than reference solution Y j (2.2.2, Method 11). (0.5 per cent);
Dissolve 2.0 g in methanol R and dilute to 10 mL with the - unspecified impumies at 298 nm: for each impurity, not
same solvent. more than the area of the peak due to impurity B in the
Related substances
(0.10 per cent);
Liquid chromatography (2.2.29). Carry ou' the ,es' avoiding
- sum of impun'ties other than B at 298 nm: not more than
exposure to actinic light. AU materials in direct cqntact with the
the area of the peak due to impurity B in the
substance to be examined like column materials, reagents, solvents,
esc. should contain only very /()W amounts of extractable metal (0.5 per cent);
cations.
- disregard limit at 298 nm: 0.5 times the area of the peak
Solven, mixture acetonitrile R, mobile phase (10:90 VIV). due to impurity B in the chromatogram obtained with
Testsolution Dissolve 30.0 mg of me substance to be reference solution (c) (0.05 per cent).
examined in 15 mL of the solvent mixture, using sonication Loss on drying (2.2.32)
if necessary, and dilute to 20.0 mL with the solventmixture. Maximum 1.5 per cent, determined on 1.000 g by drying in
Reference solulion (a) Dissolve 15.0 mg of cidopirox vacuo at 60°C.
impurityA CRS and 15.0 mg of cidopirox impurity B CRS in
the solvent mixture and dilute to 10.0 mL with the solvent Maximum 0.1 per cent) determined on 1.0 g.
mixture.
Reference sduuon (b) Dilute 1.0 mL of reference solution (a) ASSAY
to 200.0 mL with the solvent mixture. Dissolve 0.150 g in 20 mL of methanol R. Add 20 mL of
water R and titrate with 0.1 M sodium hydroxide, determining
the end-point potentiometrically (2.2.20). Carry out a blank
titration.
Reference solution (d) Mix 5 mL of reference solution (a)
and 5 mL of the test solution.
C12H17N02'
Column:
- size: I = 0.08 m, 0 = 4 mm; STORAGE
- stationary phase: diisopropylcyanosilyl silica gelfor Protected from light.
chromatography R (5 ~rh). IMPURITIES
In order to ensure desorption of interfering metal ions, every Specified impuritWs A, B, C.
new column is to be rinsed with the rinsing solutionover a
period of not less than 15 h and then with the mobile phase
for not less than 5 h at a flow rate of 0.2 mIJrnin.
Rinsing solution glacial acetic add R, acetylacetone R, andenanUomer
aawnitrilefor chromatography R, waterfor chromatography R
(0.1:0.1:50:50 VIVIVIV).
Mobilephase glacial acetic acidR, acetom·tri!e for
chromatography R, 0.96 gIL solution of sodium <derate R A. [(5RS)- 3-cyclohexyl-5-methyl-4,5-dihydco-I,2-oxazol-5-yl]
(0.01:23:71 VIVIV). acetic acid)
F/Qw rate 0.7 mUmin.
Detection Spectrophotometer at 220 nm and at 298 run.
Qo"",o
Injection 10 ~L of the test solution and reference
solutions (b), (c) and (d); inject the solvent mixture as a
blank.
y
CH,
Run lime 2.5 times the retention time of cicloplrox.
Retention lime Ciclopirox e 8 min to 11 min; if necessary
adjust the ratio of the 0.96 gIL solution of sodium edetate to B. 6-cyclohexyl-4-methyl-2H-pyran-2-one,
acetonitrile in the mobile phase.
Relative retention With reference (0 ciclopirox:
impurity A = about 0.5; impurity C = about 0,9j
System suirability At 298 nm:
- resoluu'on: minimum 2.0 between the peaks due to
ciclopirox and impurity B in the chromatogram obtained
with reference solution (d); C. 6-cyciohexyl-4-methyJpyridin-2(1H)-one.
- symmetry factor: maximum 2.0 for the principal peak in _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
the chromatogram obtainedwith the test solution.
Limits:
- impun"ty A at 220 nm: not more than the area of the
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2022 Ciclopirox Olamine 1-571
Ciclopirox Olamine *** principal spot in the chromatogram obtained with the
*** *** reference solution.
(ph. Eur. monograph 1302) *** Detection B Spray 1 plate with/em'c chloride solution R3.
av
,H with the test solution is similar in position, colourand size to
N 0 the principal spot in the chromatogram obtained with the
reference solution.
Detection C Spray the 2nd second plate with ninhydn'n
'""
CH, solution R. Heat at 110°C until the spots appear.
Results C The principal spot in the chromatogram obtained
268.4 41621-49-2 with the test solution is similar in position,colourand size to
Action and use reference solution.
Antifungal. TESTS
PhE" _ Appearance of solution
DEFINITION than reference solution BY7 (2.2.2., Method If).
6-Cyclohexyl-I-hydroxy-4-methylpyridin-2(1H)-one and
Dissolve 2.0 g in methanol R and dilute to 20 mL with the
2-aminoethanol.
same solvent.
Content
pH (2.2.3)
- cidopircx (C 12H17N02; iWr 207.3): 76.0 per cent to
8.0 to 9.0.
78.5 per cent (dried substance):
- 2-aminoethanol (C ZH7NO; lWr 61.1); 22.2 per cent to Dissolve 1.0 g in carbon dioxide-free warer R and dilute to
23.3 per cent (dried substance). 100 mL with the same solvent.
Related substances
CHARACTERS
Liquid chromatography (2.2.29). Carry out the test aw iding
Appearance
exposure to actinic light. All materials in direct contact with the
White or pale yellow, crystalline powder.
substance to be examined, such as column mauriats, reagents,
Solubility solvents, etc. should contain only smallamounts of extractable
Sparingly soluble in water, very soluble in ethanol metal CQIWns.
(96 per cent) and in methylene chloride, slightly soluble in Solvent mixture acetonitrile R, mobile phase (10:90 VIV).
ethyl acetate, practically insoluble in cyclohexane.
It shows polymorphism (5.9). examined (corresponding to about 30 mg of cidopirox) in a
IDENTIFICATION mixture of 20 ~L of anhydrous acetic acidR, 2 mL of
First idendfication: A. acetonitrile R, and 15 mL of the mobile phase, using
Second identification: B. sonication if necessary. Dilute the solution to 20.0 mL with
the mobile phase.
Reference solution (a) Dissolve 15.0 mg of cidopirox
Comparison cidopirox olamine CRS.
impurity A CRS and 15.0 mg of dcJopirox impuniy B CRS in a
If the spectra obtained in the solid stateshow differences) mixture of 1 mL of au"miuile Rand 7 mL of the mobile
dissolve the substance to be examined and the reference phase, and dilute to 10.0 mL with the mobile phase.
substance separately in the minimum volume of ethyl
aalllte R, evaporate to dryness on a water-bath and record
Reference solution (e) Dilute 2.0 mL of reference solution (b)
B. Thin-layer chromatograpby (2.2.27).
Reference solution (d) Mix 5 mL of reference solution (a)
and 5 mL of the test solution.
solvent.
Column:
Reference solution Dissolve 25 mg of cidopirox olamine CRS
- size: 1= 0.08 m, 0 = 4 mm;
- stationary phase: base-deaaiuaud end-capped cyanosilyl silita
Ptase TLC silica gel F m plale R. gel/or chromatography R (5 pm).
Pretreatment Beforeuse, predevelop 2 plates with the mobile In order to ensure desorption of interfering metal ions, every
phase until the solvent front has migrated to the top of the new column is to be rinsed with the rinsing solution over a
plates. Allow to dry in air for 5 min. period of not Jess than 15 h and then with the mobile phase
Mobile phase concentrated ammonia R, water R, anhydrow for not less than 5 h at a flow rateof 0.2 mUmin.
ethanol R (10:15:75 VIVIV). Rinsing solution acetylaceume R, anhydrous acetic acidR,
Application I 0 ~L. acetonitrile for chromatography R, waterfor chromatographY R
Development Over 213 of the plate. (0.1:0.1:50:50 VIVIVIV).
Drying In airfor 10 min. i\;lobile phase anhydrous acetic acid R, acetonitrile for
Detection A Examine in ultraviolet light at 254 om. chromatography R, 0.96 g/L solutionof sodium edetate R
(0.01:23:77 VIVIV).
Results A The principal spot in the chromatogram obtained
with the test solution is similar in positionand size to the Flew rale 0.7 mUmin.
Detection Spectrophotometer at 220 nm and at 298 run.
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1-572 Ciclosporin 2022
Injection 10 j.lL of the test solution and reference

Run time 2.5 times the retention lime of ciclopirox.
Retention time Cicfopirox e 8 min to 11 min; if necessary
adjust the ratio of the 0.96 gIL solution of sodium edetate to A. [(5RS) -j-cyclohexyl-f-methyl-d,5-dihydro-1,2-oxazol-5-yl]
acetonitrile in the mobile phase. acetic acid,
Relative retention With reference (0 ciclopirox:
impurity A == about 0.5; impurity C = about 0.9;
QO-...,pO
System suitability At 298 run:
- resolution: minimum of 2.0 between the peaks due to
impurity Band ciclopirox in the chromatogram obtained
Y CH,
with reference solution (d): B. 6-cyclohexyl-4-methyl-2H-pyran-2-one,

- symmetry [actor: maximum 2.0 for the principal peak in
Q~-...,pO
the chromatogram obtained with the test solution.
Limits:
- impurity A at 220 nm: not more than the area of the
- impurities B, Cat 298 nm: for each impurity, not more
Y CH,
than the area of me peak due to impurity B in the C.6-<:yclohexyl-4-methylpyridin-2(1H)-one.

chromatogram obtainedwith reference solution (b) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ fM"
(0.5 per cent);
- unspecified impurities at 298 nm: for each impurity, not
more than the area of the peak due to impurity B in the
Ciclosporin ***
(0.10 per cent); *** ***
- sum of impurities otherthan B at 298 nm: not more than (Ph. Eur. monograph 0994) ***
the area of the peak due to impurity B in the
-
(0.5 per cent);
disregard limit at 298 nm: 0.5 times the area of the peak
due [Q impurity B in the chromatogram obtained with
[ I
A1a -- o-AJa-- MeLeu-
MeLeu-Val-MeLeu-MeGly-AbU
II 7
-iX\H.,
MeLeu-- Mev al - N
,cH3
0HOH
i
H .CH:J
Loss on drying (2.2.32) CH,

VlUUO at a pressure not exceeding 0.1 kPa.
1203 59865-13-3
Maximum 0.1 per cent, determined on 1.0 g. Action and use
ASSAY
Calcineurin inhibitor; immunosuppressant.
2-AmlnoethanoI Preparations
Dissolve 0.250 g in 25 mL of anhydrous acetic acid R. Titrate Ciclosporin Capsules
with 0.1 M perchloric acid, detenuining the end-point Ciclosporin Eye Drops
potentiometrically (2.2.20). Ciclosporin Oral Solution
I mL of 0.1 M perchloric acid is equivalent to 6.108 mg of Ciclosporin Sterile Concentrate
CZH7NO.
!'hE" _
Clclopirox
Dissolve 0.200 g in 2 mL of methanol R. Add 38 mL of DEFINITION
water R, swirl and titrate immediately with 0.1 M sodium 1,II-Anbydro[L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-
hydroxide, determining the end-pointpotentiometrically L-Ieucyl-N-methyl-L-valyl-(ZS,3R,4R,6E)-3-hydroxy-4-
(2.2.2{}J. Carry out a blank titration. methyl-2-(methylamino)oct-6-enoyl-(2S)-2-aminobutanoyl-
Use 0.1 M sodium hydroxide, the titre of which has been N-methylglycyl-N-methyl-L-Ieucyl-L-valyl-N-methyl-L-
determined under the conditionsprescribed above using leucine] (ciclosporin A).
0.100 g of benzoic acid RV. Substance produced by Beauveria niuea (O.Rostr.) Arx
I mL of O. 1 M sodium hydroxide is equivalent to 20.73 mg of (Tolypodadium ;njlarum W.Gams) or related species.
C12H17NOz. Content
Speafred impuniies A, B, C. White or almost white powder.
Solubility
ethanol and in methylene chloride.
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2022 Ciclosporin 1-573
IDENTIFICATION - peak-to-valley ratio: minimum 2.0, where H p =

height
A. Infrared absorption spectrophotometry (2.2.24). above the baseline of the peak due to impurity D and
Comparison cidosporin CRS. H; = height above the baseline of the lowest point of the
ciclosporin; if necessary, adjust the ratio of
Results The principal peak in the chromatogram obtained Lf-dimethylethyl methyl ether to acetonitrile in the
with the test solution is similar in retention time to the mobile phase.
principal peak in the chromatogram obtained with reference
solution (a).
- for each impurity, use the concentration of ciclosporin in
TESTS reference solution (b).
Appearance of solution Limits:
The solution is clear (2.2.1) and not more intensely coloured - sum of impurities Band E: maximum 0.5 per cent;
than reference solution Yj ) BYj or R1 (2.2.2) Method If). - impun"ty G: maximum 0.4 per cent;
Dissolve 1.5 g in anhydrous ethanol R and dilute to IS mL - any otherimpun'ty: for each impurity, maximum
with me same solvent 0.3 per cent;
Specific optical rotation (2.2.7) - total: maximum 1.5 per cent;
-193 to -185 (dried substance). - reporting threshold: 0.05 per cent.
Dissolve 0.125 g in methanol R and dilute to 25.0 mL with Los. on drying (2.2.32)
the same solvent. Maximwn 2.0 per cent, determined on 1.000 g at 60 °C at a
pressure nor exceeding 0.6 kPa for 3 h.
Related substances
liquid chtomatography (2.2.29). Bacterial endotoxin. (2.6.14)
SoIwn, mixture acetonitrile R, waterR (50:50 VW). Less man 0.84 ID/mg, if intended for use in the manufacture
procedure for the removal of bacterial endotoxins. Dissolve
50 mg of the substance to be examined in a mixture of
280 mg of ethanol (96 per cenV Rand 650 mg of
Reference sdution (a) Dissolve 30.0 mg of cidasporin CRS in poIyoxyethylated castar ail R and dilute to the required
the solvent mixture and dilute to 25.0 mL with the solvent concentration using water for BET.
mixture.
ASSAY
Reference solutian (b) Dilute 2.0 mL of reference solution (a)
Reference sduiion (c) Dissolve the contents of a vial of
cidosporin for systemsuitability A CRS (containing
impurities B, D, E and G) in 5 mL of the mobile phase. System suitability Reference solution (a):
- repeatabilir;y: maximum relative standard deviation of
Column:
- size: 1= 0.25 m, 0 4.6 mm;= Calculate the percentage content of C62HIIINIIOl2 taking
- stationary phase: end-capped ac,adecylsilyl silica gelfor
chromatography R (3 ~m); into account the assigned content of cidosporin CRS.
- temperature: 80 "C: STORAGE
- pre-column heater: 80 °C; alternatively, the column is In an airtight container, protected from light. If the substance
connected to the injection port by a steel capillary rube is sterile, the container is also sterile and tamper-evident.
about 1 m long, having an internal diameter of 0.25 mm
and maintained at 80 "C; IMPURITIES
- post-column cooler: 25 "C. Specified impurities B, E, G.
Mobile phase phasphoric acid R, l,l-dimethylethyl methyl Otherdetectable impurities (thefollowing substances would, if
ether R, acetonitrile Rt, WQter for chromawgraphy R present at a sufficientleveJ, be detected by oneor otherof the tests
(0.1:5:43:52 VIVIVW). in the monograph. They are limited by the general acceptance
oitenonfar otherhmspecified impurities. It is therefore not
necessary to "identify these impun·ties for demonstration of
Detection Spectrophotometer at 210 om. complianu. See also 5.10. Conrrol of impurities in substances for
Injection 20 j.tL of the test solution and reference pharmaceutical use) C, D, F.
Run time 1.7 times the retention time of cic1osporin. pH3
[~a-o-Ala-Meleu-MeLeu-Meval-N ...HH ...CH3
ldenufication of impwides Use the chromatogram supplied
with ciclasparin for systemsuitability A CRS and the Meleu-val-Meleu-MeGly-AbuJ)(\ -
chromatogram obtained with reference solution (c) to identify II 7 ~HrOH~
the peaks due to impurities B + E, D and G.
CH,
Relativeretention With reference to ciclosporin (retention
= =
time about 26 min): impurity D about 0.9; impurities B
and E = about 1.3; impurity G about 104.= B. 1,II-aohydro[L-a1aoyl-D-a1aoyl-N-methyl-L-leucyl-N-
methyl-L-leucyl-N-methyl-L-valyl-(2S,3R,4R)-3-hydroxy-4-
methyl-2-(methylamino)octaooyl-(2S)-2-aminobutaooyl-
- retention time: ciclosporin = 25 min to 30 min;
N-methylglycyl-N-methyl-L-Ieucyl-L-valyl-N-methyt-L-
if necessary, adjust the ratio of acetonitrile to water in the
leucine] (dihydrociclosporin A),
mobile phase;
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1-574 Cilastatin Sodium 2022
H?H 3
Cilastatin Sodium
A1S_ f>AIa-- MeLeU- Met eu- M9Vsl$ O .
II H
[
Maleu-Val-MeLeu-MeGIy-Abu -; ",CH3 CH3 (ph. Eur. monograph 1408)
a ° H ~
C. 0'.1, ll-anhydro[(2S,3R,4R,6E)-3-hydroxy-4-methyl-2-
(methylamino)oet-6-enoyl-(2S)-2-aminobutanoyl-N-
.
HD,C'X"
,
HNH2
S
~co,NaCH3
HN HV-
If
..
CH,
methylglycyl-N-methyl-L-Ieucyl-L-valyl-N-methyl-L-leucyl-
L-a1anyl-o-alanyl-N-methyl-L-Ieucyl-N-methyl-L-leucyl-N-
°
methyl-L-valine] (isociclosporin A), 380.4 81129-83-1
pH) Action and use

r~a-O-Aia-MeLeu-MeLeu-Maval-N H c~ Dehydropeptidase-I inhibitor; inhibition of the renal
L Leu-Val-MeLeu-MeGIy -Abu
II 1
~'0HOH
/
metabolism of imipenem.
Preparation
Cilastatin and Imipenemfor Infusion
CH,
PhE" _
D.I,II-anhydro[L-a1anyl-o-a1anyl-N-methyl-L-leucyl-N- DEFINITION
methyl-L-leucyl-N-methyl-L-valyl-(2S,3R,4R,6E)-3- Sodium (Z}-7-[[(2R)-2-amino-2-carboxyethyl)sulfanyl]-2-
hydroxy-4-methyl-2-(methylamino)oct-6-enoyl-(2S)-2- [[[( IS)-2 ,2-dimethylcyclopropyl]carbonyl]amino]hept-2-
aminobutanoyl-N-methylglycyl-N-methyl-L-leucyl-L-valyl- enoate.
L-Ieucine] ([Leu1l]ciclosporin A, cic1osporin U), Content
pH)
Abu- o-AJa - MeLeU- Mel eU- MeV81- N HH CH, CHARACTERS
[ "'
MeLeu-Val-Meleu-MeGIy-Abu
~'f
/
Appearance
White or light yellow, amorphous, hygroscopic powder.
. 6 OH OH
Solubility
CH, Very soluble in waterand in methanol, slightly soluble in
anhydrous ethanol, very slightly soluble in dimethyl sulfoxide,
E. 1,II-anhydro[o-a1anyl-N-methyl-L-leucyl-N-methyl-L- practically insoluble in acetone and in methylene cWoride.
leucyl-N-methyl-L-valyl-(2S,3R,4R,6E)-3-hydroxy-4- IDENTIFICATION
methyl-2-(methylamino)oct-6-enoyl-(2S)-2- A. Specific optical rotation (see Tests).
aminobutanoyl-N-methylglycyl-N-methyl-L-leucyl-L-valyl-
N-methyl-L-Ieucyl-(2S)-2-aminobutanoic acid] ([Abu'] B. Infrared absorption spectrophotometry (2.2.24).
ciclosporin A, ciclosporin V), Comparison cilasrarin sodium CRS.
C. It gives reaction (a) of sodium (2.3.1).
pH)
Aia - o-AJa - MaLeu - MeLeu - Mev al - N HH .c~
TESTS
[ ,
MeLeu-- Abu-- Meleu -- MeGIy-- Abu
~.,
.I
Solution S
11 7 100 mL with the same solvent.
0HOH
CH, Appearance of solution

than reference solution Y6 (2.2.2, Metlwd II).
F. 1,ll-anhydro[L-a1anyl-o-a1anyl-N-methyl-L-leucyl-N-
methyl-L-leucyl-N-methyl-L-valyl-(2S,3R,4R,6E)-3- pH (2.2.3)
hydroxy-4-methyl-2-(methylamino)oet-6-enoyl-(2S)-2- 6.5 to 7.5 for solution S.
aminobutanoyl-N-methylglycyl-N-methyl-L-leucyl-(2S)-2- Specillc optical rotation (2.2.7)
aminobutanoyl-N-methyl-L-Ieucine] ([Abu'''] + 41.5 to + 44.5 (anhydrous substance).
ciclosporin A), Dissolve 0.250 g in a mixture of I volume of hydrochloric
, acid Rand 120 volumes of methanol R, then dilute to
Aia - o-AJa - MeLeu- MeLeu- Mev al - N
p"HH pH3
25.0 mL with the same mixture of solvents.
[ , ~., Related substances
MeLeu-Val-Meleu-MeGty-Val / Liquid chromatography (2.2.29). Prepare the solutions
11 1 OH OH 1 immediately beforeuse.
CH, Test solution Dissolve 32 mg of the substance to be
G. 1,II-anhydro[L-a1anyl-o-a1anyl-N-methyl-L-leucyl-N- solvent.
methyl-L-Ieucyl-N-methyl-L-valyl-(2S,3R,4R,6E)-3- Reference solution (a) Dilute 1.0 rnL of the test solution to
hydroxy-4-methyl-2-(methylamino)oet-6-enoyl-L-valyl-N- 100.0 mL with water R. Dilute 1.0 mL of this solution to
methylglycyl-N-methyl-L-Ieucyl-L-valyl-N-methyl-L- 10.0 rnL with water R.
leucine] ([Val7]ciclosporin A, ciclosporin D). Reference solution (b) Dissolve 3 mg of dlastatinfor system
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" suitability 1 CRS (containing impurities A, BJ E, F, G
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2022 Cilastatin Sodium 1-575
(epimer 2) and H) in water R and dilute to 2.0 mL with the =

impurity C 1.3; impurity E 3.3; impurity G =
same solvent. (epimer 1) and impurity G (epimer 2) = 1.6.
Reference sdution (c) Dissolve 3 mg of cilasuuin for system Limits:
suitability 2 CRS (containing impurities C and G (epimer 1)) - impurities A (sum of the epimers): maximum 0.5 per cent;
in waterR and dilute to 2.0 mL with the same solvent. - impurity C: maximum 0.4 per cent;
Reference solution (d) Dissolve 32 mg of mesityl oxide R - impurities E: maximum 0.3 per cent;
(impurity D) in 100.0 mL of water R. Dilute 1.0 mL of the - impun·u·es B, F, H: for each impurity, maximum
solution to 50.0 mL with water R. 0.1 per cent;
Column: - impun"ty G: for each epimer, maximum 0.1 per cent;
=
- size: 1= 0.25 rn, (2) 4.6 mmj - unspecified impurilies: for each impurity, maximum
- stationary phase: end-capped oetadecylsilyl Sl1ica gelfor 0.05 per cent;
chromatography compatible WJih lOOper centaqueous mobile - toUt!: maximum 1.0 per cent;
phases R (5 ~m); - reporting threshold: 0.03 per cent; disregard any peak due
- temperature: 50°C. to impurity D in the chromatogram obtained with
Mobile phase:
- mobile phase A: phosphate buffer solllOon pH 3.25 R; Impurity D, acetone and methanol
- mobile phase B: acetonitrile Rl, phosphate bufftrsolution Gas chromatography (2.2.28).
pH 3.25 R (50:50 VIII); Internal standard solution Dissolve 0.5 mL of propanol R in
water R and dilute to 1000 mL with the same solvent.
Time MobUe phose A Mobile pbBSC B
(min) (per cent JlIII) (per cent Vm examined in waterR, add 2.0 mL of the internal standard
0-3 100 0
solution and dilute to JO.O mL with water R.
3 - 28 100 ..... 90 0 ..... to
Reference solution Dissolve 2.0 mL of acelQne R, 0.5 mL of
28 - 38 90 10
10 ..... 50
methanol Rand 0.5 mL of mesityl oxide R (impurity D) in
38 - 63 90 ..... 50
63- 78 50 70
waler R and dilute to 1000 mL with the same solvent.
50 ..... 30 -->
To 2.0 mL of this solution add 2.0 mL of the internal
78·88 30 70
standard solution and dilute to 10.0 mL with Waler R. TIlls
solution contains 3J6 J.1g of acetone, 79 ug of methanol and
86 ~g of impurity D per millilitre.
Cdumn:
Injection 20~. - material: fused silica;
Identification of impurities Use the chromatogram supplied =
- size: I 30 m, 0 0.53 mm; =
with cilastatin for system suitability 1 CRS and the - stationary phase: macrogol20 000 R (film thickness
chromatogram obtained with reference solution (b) to 1.0 urn).
identify the peaks due to impurities A, B, E, F, G (epimer 2) Carrier gas helium for chromatography R.
and H; use the chromatogram supplied with cilastatin for
Flow rale 9 mUmin.
system suitability 2 CRS and the chromatogram obtained with
reference solution (c) to identify the peaks due to impurities Temperature:
C and G (epimer 1); use the chromatogram obtained with
reference solution (d) to identify the peak due to impurity D. Time Temperature
(min) Cq
Relative retention With reference to cilastatin (retention
Column 0-2.5 50
time = about 50 min): impurity E = about 0.2; impurity A 2.5 • 5 50 -> 70
(epimer 1) = about 0.60; impurity A
5 - 5.5 70
{epimer 2) = about 0.62; impurity D = about 0.9;
Injection port 160
impurity F = about 0.98; impurity G (epimer 1) = about
Detector 220
=
1.02; impurity G (epimer 2) about 1.05;
impurity H = about 1.06; impurity B = about 1.17;
=
impurity C about 1.23. Detection Flame ionisation.
System suitabIlity: Injection I~L.
=
- peak-to-oalley ratio: minimum 10, where Hp height Calculate the percentage contents of acetone, methanol and
above the baseline of the peak due to impurity F and impurity D using the following expression:
HI} = height above the baseline of the lowest point of the
curve separating this peak from the peak due to cilastatin
in the chromatogram obtained with reference solution (b);
=
- peak-to-vaOey ratrO: minimum 2.5, where Hp height
above the baseline of the peak due to impurity G C concentration of the solvent in the reference solution, in IlgfmL;
W quantity of cjfasrarinsodium in the test solution, in milligrams;
(epimer 1) and H; = height above the baseline of the R" ratio of the area of the solvent peak 10 the area of the propanol
lowest point of the curve separating this peak from the peak in the chromatogram obtained with the lest solution;
peak due to cilastatin in the chromatogram obtained with ~ ratio of the area of the solvent peak [0 the area of the propanol
reference solution (c). peak in the chromatogram obtained with the reference solution.

Limits:
- for aU impurities, use the concentration of cllastatin
- acetone: maximum 1.0 per cent mlm;
sodium in reference solution (a);
- methanol: maximum 0.5 per cent mtm;
- impun·'Y D: maximum 0.4 per cent mlm.
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1-576 Cilazapril 2022
Water (2.5.12)
ASSAY F. (Z)-7-[[(ZR)-Z-amino-Z-carboxyethyl]sulfanyl]-Z-[(Z,3-
Dissolve 0.100 g in 30 mL of melhanol R and add 5 mL of dimethylbut-3-enoyl)amino]hept-2-enoic acid,
water R. Add 0.1 M hydrochloric acid to a pH of about 3.0.
Carry our a potentiometric titration (2.2.20), using 0.1 M HO:!C~S~CO:!H
sodium hydroxide. Three jumps of potential are observed.
Read the volume added between the 1st and the 3rd point of
1\ 2
HNH - - H>v--'
"' N .
H
I"
CH
CH,
inflexion. and eplmerat C" 0
1 mL of 0.1 M sodium hydroxide is equivalent to 19.0Z mg
of C,Jl2,N2Na05S, G. (EJ-(ZRS)-7-[[(ZR)-Z-amino-Z-carboxyethyl]sulfanyI]-Z-
[[[(I S)-Z,Z-<limethylcyclopropyl]carbonyl] amino [hept-3-
STORAGE
enoic acid,
In an airtight container, at a temperature of 2 °C to 8°C.
If the substance is sterile, store in a sterile, airtight, tamper-
evidentcontainer.
IMPURITIES
Specified impuriues A, B, C, D, E, F, 0, H.
HO,c~S~co,H CH,
H. (Z)-7 -[(Z-aminoethyl)sulfanyl)-Z-[[[(IS)-2,2-
1\
H NH
It I ._
HN 20
H>v-- dimethylcyclopropyl]carbonyl)amino]hepr-2-enoic acid.
I' CH,
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
and eplmeral S 0
A. (Z)-7-[(RS)-[(ZR)-Z-amino-Z-carboxyethyl]sulfinyl]-Z-
[[[(1S)-Z, Z-<limethylcyclopropyl]carbonyl] amino] hept- Z- Cilazapril
enoic acid, .
H""C-.....,/"'S~co,H CH
1\
HNH
- - I
HN.
H>v--'
r4 :n" CH, 1<,0
H3C--{ H CH3 0
o and epln'l8r at C"
B. (Z)-7 -Il(ZR)-2-carboxy-2- [[( IRS)-I-methyl-3-oxobutyl] 92077-78-6

amino]ethyl]sulfanyl]-Z-[[[(IS)-Z,Z-<limethylcyclopropyl]
carbonyl]amino]hept-2-enoic acid, Action and use
Angiotensin converting enzyme inhibitor.
PbE<I _
DEFINITION
( IS,9S)-9-[[(IS) -l-(EthoxycarbonyI)-3-phenylpropyl]arnino]-
I 0-oxooetahydro-6H-pyridazino[I,2-a] [I ,Z]dia2Cpine-l-
carboxylic acid monohydrate.
C. (Z)-7 -[[(ZR)-Z-carboxy-Z-[( 1,I-<limethyl-3-oxobutyl) Content
amino]ethyl)sulfanyl]-Z-[[[(IS)-Z,Z-<limetbylcyclopropyl] 98.5 per cent to 101.5 per cent (anhydrous substance).
carbonyl]amino]hept-2-enoic acid,
CHARACTERS
Appearance
Solublliry
Slightly soluble in water, freely soluble in methanol and in
methylene chloride.
D. 4-methylpent-3-en-Z-one (mesityl oxide),
IDENTIFICATION
Ho,CTS~co,H
Comparison ci1azapril CRS.
H NI<, 0
B. Specific optical rotation (see Tests).
E. 7-[[(ZR)-Z-amino-Z-carboxyethyl]sulfanyl]-Z-oxoheptanoic TESTS
acid, Specific optical rotation (2.2.7)
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2022 CiiazapriI 1-577
Dissolve 0.200 gino. 067 M phosphate buffer solution - symmetry factor. maximum 3.0 for the peak due to
pH 7.0 R, with the aid of ultrasound if necessary J and dilute cilazapril.
to 50.0 mL with the same buffer solution. Carry out the Limits:
determination at 365 om. - impurityB: not more than the area of the principal peak in
Impurity A the chromatogram obtained with reference solution (a)
Thin-layer chromatography (2.2.27). (0.5 per cent);
Test solution Dissolve 0.20 g of the substance to be - impuniy D: not more than 0.4 times the area of the
examined in methanol R and dilute to 5.0 mL with the same principal peak in the chromatogram obtained with
solvent. reference solution (a) (0.2 per cent);
Reference solution (a) Dissolve 2 mg of cilazapril - impun·cy C: not more than 0.2 times the area of the
impun'ty A CRS in me/hanoi R and dilute to 50.0 mL with the
same solvent.
- unspecified impuniies: for each impurity, not more than
Reference solution (b) Dissolve 5 mg of cilazapri/ 0.2 times the area of the principal peak in the
impurity A CRS and 5 mg of the substance to be examined in chromatogram obtained with reference solution (a)
methanol R and dilute to 10.0 mL with the same solvent. (0.10 per cent);
Ptote TLC silica gelplateR. - total: not more than twice the area of the principal peak in
A10bile phase glacial acetic acidR, waterR, hexane R, the chromatogram obtained with reference solution (a)
me/hanoi R, ethyl a<e/ate R (5:5:15:15:60 VIVIVIVIJI). (I per cent);
Application 5 ~L. - disregard limit: 0.1 times the area of the principal peak in
(0.05 per cent), disregard any peak due [0 impurity A.
Drying In a current of cold air for 10 min.
Water (2.5.12)
Detection Spray with a freshly prepared mixture of 1 volume 3.5 per cent to 5.0 per cent, determined on 0.300 g.
of potassium iodobisnnuhate soluei<m R and 10 volumes of dilute
auric acidR and then with dilute hydrogen peroxide solution R. Sulfated ash (2.4.14)
- the chromatogram shows 2 dearly separated spots. ASSAY
Limit: Dissolve 0.300 g in 10 mL of anhydrous ethanol R and add
- impun·ty A: any spot due to impurity A is not more 50 mL of waterR. Titrate with 0.1 M sodium hydroxide,
intense than the corresponding spot in the chromatogram determining the end-point potentiometrically (2.2.21J). Carry
obtained with reference solution (a) (0.1 per cent). out a blank titration.
Related substances I mL of 0.1 M sodium hydroxide is equivalent to 41.75 mg of
Liquid chromatography (2.2.29). C22H31N305'
Test solution Dissolve 25.0 mg of the substance to be STORAGE

examined in the mobile phase and dilute to 50.0 mL with Protected from light.
Reference solution (aJ Dilute 1.0 mL of the test solution to Specified impurities A, B, C, D.
solution to 20.0 rnL with the mobile phase.
Reference solution (b) Dissolve 5.0 mg of Cl7azapril
impun·ty D CRS in the test solution and dilute to 10.0 mL
with the test solution.
Column:
- size: I;;;; 0.25 m, 0 ;;;; 4.6 nun;
- stationary phase: IXtade<y/silyl silica gelfor chromatography R A. I, I-dimethylethyl (IS,9S)-9-[[(S)-I-(ethoxycarbonyl)-3-
(5 urn). phenylpropyljaminoj-10-oxooctahydro-6H-pyridazino[I,2-
a) [1,2)diazepine-I-carboxylate,
JHobi/e phase Mix 10 volumes of tn"ethylamine Rand
750 volumes of waterR, adjust to pH 2.30 with phosphon'c
acid R, and add 200 volumes of tetrahydrofuran R.
Injection 20 ~L.
Run time Twice the retention time of cilazapri1; when
impurity A is present, it may be necessary to continue the B. (I S,9S)-9-[[(S)-I -carboxy-S-phenylpropyljamino)-10-
chromatography until it is eluted. oxooctahydro-6H-pyridazino[I,2-a) [1,2)diazepine-l-
carboxylic acid,
Rdatioe retention With reference to cilazapriJ:
impurity B = about 0.6; impurity D = about 0.9;
=
impurity C about 1.6; impurity A 4 to 5.=
impurity D and cilazapril;
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1-578 Cimetidine 2022

Comparison cimetidine CRS.
substance separately in 2-propanol R, evaporate to dryness
C. ethyl (lS,9S)-9-[[(S)-I-(ethoxycarbonyl)-3-phenylpropyIJ and recordnew spectra using the residues.
amino)-10-oxooctahydro-6H-pyridazino[I,2-aJ [1,2J C. Thin-layer chromatography (2.2.27).
diazepine-l-carboxylate, Test solution Dissolve 10 mg of the substance to be
solvent.
Reference solution Dissolve 10 rngof cimetidine CRS in
Plate TLC silica gel GFm plateR.
MobIle phase concentrated ammonia RJ methanol R, ethyl
D. (IS,9S)-9-[[(R)-I-(ethoxycarbonyl)-3-
aatate R (15:20:65 VIV/JI).
phenylpropylJamino)-10-oxooctahydro-6H-pyridazino[I,2-
aJ[1,2)diazepine-I-carboxylic acid. Application 5 ~L.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE.- Development Over 3/4 of the plate.
Drying In a current of cold ale.
Detection Expose to iodine vapour until maximum contrast
has been obtained and examine in ultraviolet light at
Cimetidine *** 254 nm.
*** *** Results The principal spot in the chromatogram obtained
(ph. Eur. monograph 0756) *** with the test solution is similar in positionand size to the
reference solution.
TESTS
than reference solution Y, (2.2.2, Me/hod If).
252.3 51481-61-9 Dissolve 3.0 g in 12 mL of 1 M hydrochloric acid and dilute
Action and use
HistamineH2 receptor antagonist; treatment of peptic Related substances
ulceration. Liquid chromatography (2.2.2!l).
Preparadons Test solution Dissolve 20 mg of the substance to be
Cimetidine Injection examined in mobile phaseA and dilute to 50.0 mL with
mobile phase A.
Cimetidine Oral Solution
Reference solutwn (a) Dilute 1.0 mL of the lest solution to
Cimetidine Oral Suspension
Cimetidine Tablets solution to 10.0 mL with mobile phase A.
PhE.- ~ _ Reference solution (b) Dissolve the contentsof a vial of
cimetidine for system suitability CRS (containing impurities B,
DEFINITION
C, D, E, G and H) in 1.0 mL of mobile phase A.
2-Cyano-I-methyl-3-[2-[[(5-methyl-IH-imidazol-4-yl)methyl]
sulfanyl]ethyl]guanidine. Reference solution (c) Dissolve 4 mg of cimetidine for peah
identification CRS (containing impurity F) in mobile phase A
Content and dilute to 10.0 mL with mobile phase A.
Golumn:
CHARACTERS - size: 1= 0.25 m, 0 = 4.6 rnm,
Appearance - statwnary phase: end-capped octode<ylsilyl silica gelfor
White or almost white powder. chromaUJgraphy rompatible with 100 per cent aquecus mobi/e
Solubillty phases R (5 J1IIl).
Slightly soluble in water, soluble in ethanol (96 per cent), Mobile phaseA Mix 0.4 volumes of diethylamine R and
practically insoluble in methylene chloride. It dissolves in 780 volumes of a l.l gIL solution of sodium
dilute mineral acids. hexanesulfonau R; adjust to pH 2.8 with phosphoric acid R;
It shows polymorphism (5.9). add 250 volumes of methanol R2;
Mobile phase B meshand R2;
IDENTIFICATION
First identification: B. Mobile phase A
Time Mobile phase B
Second identification: A, C. (min) (per cent VII') (per cent I'll")
A. Melting point (2.2.14): 139 °C to 144 °C. 0- 60 tOO 0
If necessary, dissolve the substance to be examined in 60 - 65 100 --+ 90 0-->10
2-propanol R, evaporate to dryness and determine the melting 65 - 120 90 to
point again.
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2022 Cimetidine 1-579

Injection SO ~1.
with cimetidine for system suitability CRS and the
chromatogram obtained with reference solution (b) to A. methyI3-cyano-I-[2-[[(S-methyl-IH-imidazol-4-yl)methyl]
identify me peaks due to impurities B, C, D, E, G and Hj sulfanyljethyljcarbamimidorhioate,
use the chromatogram supplied with cimetidine forpeak
CN
idenu]ication CRS and the chromatogram obtained with I
f=N N
reference solution (c) to identify the peak due to impurity F.
HN~S )lOCH
Relative retention With reference to cimetidine (retention U ...............N
H '
tlme e about 18 min): impurity G :::: about 0.2; H,C
impurity E :::: about 0.4; impurity D :::: about 1.5;
impurity C :::: about 1.6; impurity B :::: about 2.0; B. methyl 3-cyano-I-[2-[[(S-methyl-IH-imidazol-4-yl)methyl]
impurity H :::: about 23; impurity F :::: about 4.6. sulfanyl]ethyl]carbamimidate,
- resolution: minimum 1.5 between the peaks due [0
impurities D and C.
Limits:
- correaion factors: for the calculation of content, multiply
corresponding correction factor: impurity C = 2.5;
impurity D =3.3; impurity E =0.7; impurity G =0.6. C. 1-[(methylamino)[[2-[[(S-methyl-IH-imidazol-4-
- impuriues B, C, D, E, F, G, H: for each impurity, not yl)methyl] sulfanyl]ethyl]amino] methylidene] urea,
more than the area of the principal peak in the
chromatogram obtainedwith reference solution (a)
(0.2 per cent);
- unspecified impun'ties: for each impurity, not more than
(0.10 per cent); D.I-methyl-3-[2-[[(S-methyl-IH-imidazol-4-yl)methyl]
- total: not more than 5 times the area of the principal peak sulfanyl]ethyl]guanidine,
(1.0 per cent);
- disregard lim,i: 0.25 times the area of the principal peak in
(O.OS per cent).
Maximum 0.5 per cent, determined on 1,000 g by drying in E. 2-cyano-I-methyl-3[2-[[(S-methyl-1 H-imidazol-4-
an oven at 105 "C, yl)methyl]sulfinyl]ethyIJguanidine,
ASSAY
with 0.1 M perchlonc cum determining the end point
potentiometrically (2. 2.21!).
F. 2-cyano-I,3-bis[2-[[(S-methyl-lH-imidazol-4-yl)methyl]
1 mL of 0.1 M perchforU: acid is equivalent to 2S.23 mg of
sulfanyljethyljguanldine,
CIOH,oN,S
STORAGE
IMPURITIES
Specified impuruies BJ CJ DJ EJ F, G, H.
Other deuaable impurities (thefollowing substances would, if G" z-cyano-Lj-dimethylguanldine,
present at a sufficitmlevelJ be deteaed by one or other of the tests
in the manograph. They are limited by thegeneral acceptance
monograph Substances/or pharmaceutical use (2034). It is
therefore not neassary to identify these impurities for
demonstration of compliance. See also 5.10. Control 0/impun"ties
in subsrances for pharmaceutical use) AJ I, J.
H. 1,1'-(disulfanediyldiethylene)bis(2-cyano-3-
methylguanidine),
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1-580 Cimetidine Hydrochloride 2022
;=N Reference solution Dissolve 10 mg of cimetidine

HN~OH hydrochloride CRS in methanol R and dilute to 10 rnL with the
CH, same solvent.
Piau TLC silica gel GFm piau R.
1. (5-methyl-IH-imidazol-4-yl)methanol, Mobik phase concentrated ammonia RJ methanol RJ ethyl
aatau R (15:20:65 VIV/V).
;=N Application 5 flL.
HN,
r-l
CH,
<;»
_8
~NH, Development Over 3/4 of the plate.
Drying In a current of cold air
Detection Expose to iodine vapour until maximum contrast
J. 2-[[(5-methyl-IH-imidazol-4-yl)methyl) has been obtained and examine in ultraviolet light at
suUanyijethananline. 254 nm.
____________________ 1"''' Results The principal spot in the chromatogram obtained
reference solution.
Cimetidine Hydrochloride D. It gives reaction (a) of chlorides (2.3.1).
(Ph. Bur. monograph 1500) TESTS

than reference solution Ys (2.2.2, MethodIf).
, Hel Dissolve 3.0 g in 12 rnL of I M hydrochloric acid and dilute
pH (2.2.3)
4.0 '0 5.0.
288.8 70059-30-2 Dissolve 100 mg in carbon dioxide-jm water R and dilute to
Action and use
Histamine Hz receptor antagonist; treatment of peptic Related substances
ulceration. Liquid chromatography (2.2.29).
Preparation Test solution Dissolve 20 mg of the substance to be
CimetidineInjection examined in mobile phaseA and diluteto 50.0 mL with
mobile phase A.
PhE" ---------- Reference solution (a) Dilute 1.0 rnL of the test solution to
DEFINITION 100.0 rnL with mobile phase A. Dilute 2.0 rnL of this
2-Cyano-I-methyl-3-[2-[[(5-methyl-IH-imidazol-4-yl)methyl] solution to 10.0 mL with mobile phaseA.
sulfanyl]ethyl]guanidine hydrochloride, Reference solutian (b) Dissolve the contents of a vial of
Content cimetidine for system suitability CRS (containing impurities BJ
98.5 per cent to 101.5 per cent (dried substance). C, D, E, G and II) in 1.0 rnL of mobile phase A.
Reference solution (c) Dissolve 4 mg of cimetidine for peak
CHARACTERS
identificaliou CRS (containing impurity F) in mobile phase A
Appearance and dilute to 10.0 rnL with mobile phase A.
Column:
Solub111ty - size: 1= 0.25 m, 0 = 4.6 nun;
Freely soluble in water, sparingly soluble in anhydrous - stationary phase: end-copped octad«y/silyl silica gelfor
ethanol. chromatography compatible with 100 per centaqueous mobile
IDENTIFICATION phases R (5 pm).
First idemificauon: H, D. Mobile phaseA' Mix 0.4 volumes of diethylamine Rand
Second idenufication: A, C, D. 780 volumes of a 1.1 gIL solutionof sodium
A. Ultraviolet and visible absorption spectrophotometry hexanesulfonau R. Adjust to pH 2.8 with phosphoric acidR
(2.2.25).
and add 250 volumesof methanol R2;
Test solution Dissolve 70 mg in 0.2 M sulfuric acid and dilute Mobile phase B methanol R2;
to 100.0 rnL with the same acid. Dilute 2.0 rnL of this
TIme MobUe phase A Mobile phase B
solution to 100.0 rnL with 0.2 M sulfuric acid. (per cent VjJJ)
(min> (per cent VjJJ)
Specific absorbance at theabsorption maximum at 218 nm 0-60 100 0
650 to 705. 60 - 65 100 -+ 90 o -+ 10
B. Infrared absorption spectrophotometry (2.2.24). 65 - 120 90 10
Comparison dmetidine hydrochloride CRS.
C. Thin-layer chromatography (2.2.27). Flow rate 1.1 mUmin.
Test solution Dissolve 10 mg of the substance to be Detection Spectrophotometer at 220 nm.
examined in methanol R and dilute to 10 mL with the same Injection 50 flL.
solvent.
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2022 Cimetidine Hydrochloride 1-581

with cimetidine for system suitability CRS and the
chromatogram obtained withreference solution (b) (0
identify the peaks due to the impurities B, C, D J EJ G and
H; use the chromatogram supplied with cimetidine for peak
idemlfiauion CRS and the chromatogram obtained with A. methyI3-qano-I-[2-[[(5-methyl-IH-imidawl-4-yl)methyl]
reference solution (c) to identify the peakdue to impurity F. sulfanyl]ethyl]carbamimidothioate,
Relative retention With reference to cimetidine (retention
time = about 18 min): impurity G = about 0.2; CN
I
impurity E = about 0.4,; impurity D = about 1.5; ~N N
impurity C = about 1.6; impurity B = about 2.0; HN, ~J.s )l,
impurity H = about 2.3; impurity F = about 4.6.
H,C
t" '-"""" ~~ OCH,

impurities D and C. B. methyl 3-cyano-I-[2-[[(5-methyl-IH-imidazol-4-yl)methyl]
sulfanyl]ethyl]carbamimidate,
Limits:
- COtiUlIM factors: for the calculation of content, multiply
= =
impurity D 3.3; impurity E 0.1; impurity G 0.6; =
- impuniies B, C, D J E, F, 0, H: for each impurity, not
more than the area of the principal peakin the
(0.2 per cent); C. 1-[(methylamino)[[2-[[(5-methyl-lH-imidazol-4-
- unspecified impurities: for each impurity, not more than yl)methyl]sulfanyl]ethyl]amino]methylidene]urea,
0.5 times the area of the principal peakin me
~N NH
(0.10 per cent); HN,~ s~)l, ,CH,
- total: not more than 5 times the area of the principal peak ~c
T'-"""" ~ N
H
(1.0 per cent);
D.I-methyl-3-[2-[[(5-methyl-IH-imidazol-4-yl)methyl]
sulfanyl]ethyl]guanidine,
(0.05 per cent).
an oven at 105°C.
E. 2-cyano-I-methyl-3[2-[[(5-methyl-IH-imidazol-4-
ASSAY yl)methyl]sulfinyl]ethyl]guanidine,
Dissolve 0.200 g in a mixture of 5 mL of 0.01 M hydroch/ori<
acid and 50 mL of ethanol (96 percent) R. Carry out a
potentiometric titration (2.2.20), using 0.1 M sodium
inflexion.
I mL of 0.1 M sadium hydroxide is equivalent to 28.88 mg of
ClOH"CIN.S. F. 2-cyano-I,3-bis[2-[[(5-methyl-1 H-imidazol-4-yl)methyl]-
sulfanyl]ethyl]guanidine.
STORAGE
Protected from ligbt.
IMPURITIES
Specified impurities B, C, D, E, F, 0, H.
Otherdetectable impurities (thefol/awing substances would, if
present at a sufficient level, be deteaed by one or other of the tesu G.2-cyano-I,3-dimethylguanidine,
in the monograph. They are limited by the general acceptame
criterion for other/unspecified impun'ties and/or by the general CN
I
monograph Substances for pharmaceutical use (2034). I, is N
therefore not nuessary to idemify these impurities for
demonstration of compliance. See also 5.10. Conrrol of impurities H,C'~y~~s,s~~)l,~,CH,
in substances for phannaceutical use) A, I, J. N
I
CN
H. 1,1 '-(disulfanediyldiethylene)bis(2-cyano-3-
methylguanidine),
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1-582 Cinchocaine Hydrochloride 2022
j=N Reference solution Dissolve 20 mg of cinchocaine

HN~OH hydrochloride CRS in methanol R and dilute to 5 mL with the
CH, same solvent.
Plare TLC silica gelFZ34 plare R.
1. (5-methyl-IH-imidazol-4-yI)methanol, lYlob,le phase ammonia R, methanol R, acewne R, toluene R
(1:5:30:50 VIVIVIV).
j=N
r
HN,A
CH,
<:»:
_5
""""""'-'NH2
Appliratwn 5 ~L.
Drying In a current of warm air for 15 min.
J. 2-[[(5-methyl-IH-imidazol-4-yI)methyl] Detection Examine in ultraviolet light at 254 nm.
sulfanyqethanamine. Results The principal spot in the chromatogram obtained
____ ~ PIIE" with the test solution is similar in position and size to the
reference solution.
D. Dissolve 0.5 g in 5 mL of warer R. Add 1 mL of d.7ure
Cinchocaine Hydrochloride *** ammonia R2. A white precipitate is formed. Filter, wash the
*** *** precipitate with 5 quantities, each of 10 mL, of water Rand
(Ph. Bur. monograph 1088) *** dry in a desiccator. It melts at 64 'C to 66 'C (2.2.14).
E. It gives reaction (a) of chlorides (2.3.1).
TESTS
Solution S
HCI
Solution S is dear (2.2.1) and not more intensely coloured
C~30CIN30, 379.9 61-12-1 than reference solution Y6 (2.2.2, J'Aethod II).
Action and use pH (2.2.3)
Local anaesthetic. 5.0 to 6.0.
Dilute 10 mL of solution S to 50 mL with carbon dWxide-free
PIlE" ~ _
water R.
2-Butoxy-N-[2-(diethylamino)ethyl]quinoline-4-carboxamide Liquid chromatography (2.2.29).
hydrochloride. Solvent mixrure Mobile phase B, water R (10:90 VIV).
Content Test solution Dissolve 50.0 mg of the substance lO be
98.5 per cent to 101.0 per cent (dried substance). examined in the solvent mixture and dilute to 50.0 mL with
CHARACTERS the solventmixture.
Appearance Reference solution (a) Dilute 1.0 mL of the test solution to
White or almostwhite, crystalline powder or colourless 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
crystals, hygroscopic, very easily agglomerates. solution to 10.0 mL with the solvent mixture.
Solubillty Reference solution (b) Dissolve 2 mg of cinchocaine
Verysoluble in water, freely soluble in acetone, in ethanol impurity A CRS and 2 mg of cinrhocaine impuri.y B CRS in
(96 per cent) and in methylene chloride. the solventmixture and dilute to 100 mL with the solvent
mixture. Dilute 1 mL of the solution to 10 mL with the
IDENTIFICATION solventmixture.
Column:
Second idenujica/ion: A, C, D, E. - size: 1= 0.25 m, (2) = 3 rnrn,
A. Ultraviolet and visible absorption spectrophotometry - statwnary phase: end-capped cross-linked octaduylsiIyI silica
(2.2.25). gelfor chromatography R (5 urn);
Testsolution Dissolve 60.0 mg in a 103 gIL solution of - temperamre: 40 "C.
hydroch/orU; add R and dilute to 100 mL with the sarne acid Mobile phase:
solution. Dilute 2 mL of this solution to 100 mL with a - mobile phase A: to 950 mL of water for chroma.ography R
103 gIL solution of hydrochloric acidR. add 0.6 mL of phosphoric acid R, adjust to pH 2.0 with
Special range 220-350 urn. the same acid and dilute to 1000 mL with waterfor
Absorption maxima 246 nm and 319 nm. chromatography R;
- mobile phase B: acewnim7e for chromawgraphy R;
=
Absorbance ratio A,.. JA319 2.7 to 3.0.
Comparison cinchocaine hydrochloride CRS.
solvent.
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2022 Cineole 1-583

(min) (per cent VJlI) (per cent V/JI)
o-z 90 10
2 - 5.5 90 ...... 82 10 ..... 18
5.5 - 10.5 82 --> 50 18 --> 50
10,5 - 16.5 50 -+ 49 50 --> 51
B. 2-hydroxyquinoline-4-carboxylic acid,
16.5 - 19.5 49 --> 46 51 ..... 54
19.5 - 24.5 46 --> 30 54 --> 70
Flow rale 0.9 mUmin.

Inj«tion I0 ~L.
Relative retention With reference to cinchocaine (retention
tlme » about 10.5 min): impurity B = about 0.5;
impurity A = about 0.6. C. N-[2-(diethylamino)ethyl]-2-hydroxyquinoline-4-
carboxamide,
System suitabIlity Reference solution (b):
impurities B and A.
- for each impurity) use the concentration of cinchocaine
Limits: D. 2-hutoxyquinoline-4-carboxylic acid.
- unspecified impuniies: for each impurity, maximum _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
0.10 per cent;
Cineole ***
vaawat 60°C.
*** ***
Sulfated ash (2.4.1 f) .
Maximum 0.1 per cent, determined on 1.0 g. CH,
~CH'
ASSAY
hydrochloric acid and 50 mL of ethanol (96 per cent) R. Carry CH,
C,oH,sO 154.3 4711-82-6
inflexion.
PIlE" _
CzoH,oCIN,Oz. DEFINITION
STORAGE 1,3,3-Trimethyl-2-oxa bicycle[2.2.2] octane.
In an airtight container, protected from light. CHARACTERS
Otherdetectable impuricies (the following substanw would, if Clear colourless liquid.
present at a sufficient level, be delated by om or other of the tests Solubility
in the monograph. They a,. limited ~ the general acceptance Practically insoluble in water, miscible with alcohol and with
criterion Jorother/unspecified impurities and/or by thegeneral methylene chloride.
monograph Substanwfor pharmaceutical use (2034). It is It solidifies at about 0.5 °C.
IDENTIFICATION
demonstration of compliance. See also5.10. Control oj impuriues
in substances for pharmaceutical use) A, B, C, D. A. Refractive index (see Tests).
B. 'Thin-layer chromatography (2.2.27).
Testsolution Dilute 1 mL of solution S (see Tests) to
25 mL with akoholR.
Reference solution Mix 80 mg of cineole CRS with akohol R
and dilute to 10 mL with the same solvent.
Plate TLC silica gelplale R.
Mobile phase ethylacetale R, toluene R (10:90 VIV).
A. 2-chloro-N-[2-(diethylamino)ethyl]quinoline-4- Applkation 2~.
carboxamide, Development Over 213 of the plate.
Detection Spray with anisaldehyde solution R, heat at
100-105 °C for 5 min.
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1-584 Cinnamic Acid 2022
Results The principal spot in the chromatogram obtained £0 the internal standard, to the area of the peak due to
with the test solution is similar in position, colour and size to internal standard: this ratio is not greater than R
the principal spot in the chromatogram obtained with the (2 per cent),
reference solution. - disregard limir. 0.025 times the area of the principal peak
C. To 0.1 mL add 4 mL of su/furic acid R. An orange-red in the chromatogram obtained with reference solution (a)
colour develops. Add 0.2 mL of fonnoldehyde solution R. (0.05 per cent),
The colour changes to deep brown. Residue on evaporation
TESTS Maximum 0.1 per cent.
Solution S To 2.0 g add 5 mL of water R J evaporate to dryness on a
Dilute 2.00 g to 10.0 mL with alcohol R. water-bath and dry at 100-105 "C for I h. The residue
weighs a maximum of 2 mg.
Solution S is clear (2.2.1) and colourless (2.2.2, Method 1). STORAGE
Chlrallmpurlties In an airtight container, protected from light.
The optical rotation (2.2.7) of solution S is -0.10° to IMPURITIES
+ 0.10'.
Refractive index (2.2.6)
1.456 to 1.460.
Related substances
Gas chromatography (2.2.2lf).
Internal standardsolution Dissolve 1.0 g of camphor R in
heptane R and dilute to 200 mL with the same solvent.
Test solution (aJ Dissolve 2.5 g of the substance to he A. l-methyl-4-(I-methylethyl)-1-oxahicyclo[2.2.I]heptane
examined in heptane R and dilute to 25.0 mL with the same (1,4-cineole)..
solvent. _____________________ "'E"
Testsolution (b) Dissolve 2.5 g of the substance to be
examined in heptaneR, add 5.0 mL of the internal standard
solution and dilute to 25.0 mL with heptane R.
Reference solution (a) To 2.0 mL of test solution (a) add Cinnamic Acid
20.0 mL of the internal standard solution and dilute to
100.0 mL with heptane R. ~COOH
Reference solution (b) Dissolve 50 mg of 1,4~n",le Rand
SO mg of the substance to be examined in heptane Rand V
dilute to 50.0 mL with the same solvent.
Column: 148.2 621-81-9
- size: 1= 30 m, (2) = 0.25 mm,
- stationary phase: macrogol 20 000 R (film thickness
Action and use
Antimicrobial preservative; excipient.
0.25 pm).
Carrier gas heliumfor chromatography R. DEFINITION
Linearvelocity 45 em/s. Cinnamic Acid is (.E)-3-phenylprop-2-enoic acid. It contains
Split-ratw 1:70. not less than 99.0% and not more than 100.5% of CgH S02 ,
calculated with reference to the dried substance.
Temperature:
CHARACTERISTICS
Time Temperature Colourless crystals.
(min) Cq
Very slightly soluble in waler; freely soluble in ethanol (96%);
Column 0-10 50
10 - 35 50 - i 100 soluble in ether.
35 - 45 100 ..... 200
IDENfIFICATION
45·55 200
Injection port 220
A. The i'!frared absorption spearnm, Appendix II A, is
concordant with the reference spectrum of cinnamic acid
Detector 250
(RS 062).
B. The lightabsorption, Appendix IT B, in the range 230 to
350 urn of a 0.0010% wlv solution in O.lM sodium hydroxide
Injection I~. exhibits a maximum only at 267 run. The absorbance at
System suitability Reference solution (b): 261 urn is about 104.
- resolution: minimwn 10 between the peaks due to
TESTS
impurity A and to cineole.
Melting point
Limits: 132' to 134', Appendix V A.
- total: calculate the ratio (R) of the area of the peak due to
cineole to the area of the peak due (0 the internal Ethanol-insoluble matter
standard from the chromatogram obtained with reference A 10% wlv solution in ethanol (96%) is clear, Appendix N A.
solution (a); from the chromatogram obtained with test Related substances
solution (b), calculate the ratio of the swn of the areas of Carry out the method for thin-layer chromatography,
any peaks) apart from the principal peak and the peak due Appendix ill A, using the following solutions in methanol.
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2022 Cinnarizine 1-585
(I) 5.0% wrv of the substance being examined. IDENTIFICATION

(2) 0.025% w/v of the substance being examined. First identification: A.) B.
CHROMATOGRAPHIC CONDITIONS Second identification: A, C, D.
(a) Use as the coating silica gel GFZ54 ' A. Melting point (2.2.14): 118 -c to 122 'C.
(h) Use the mobile phase as described below. B. Infrared absorption spectrophotometry (2.2.24).
(c) Apply 5 ~L of each solution. Comparison cinnatizine CRS.
(d) Develop the plate to 15 ern. C. Thin-layer chromatography (2.2.27).
(e) After removal of the plate, dry in air and examine under Test solution Dissolve 10 mg of the substance to be
ultraviclet light (254 nm). examined in methanol R and dilute to 20 mL with the same
solvent.
MOBILE PHASE
Reference solution (a) Dissolve 10 mg of cinnanzine CRS in
10 volumes of glacial acetic acid and 90 volumesof toluene. methanol R and dilute to 20 mL with the same solvent.
LIMITS Reference solution (b) Dissolve 10 mg of cinnasisine CRS and
Any secondary spot in die chromatogram obtained with 10 mg ofjlunarizine dihydrochloride CRS in methanol Rand
solution (1) is not more intense than the spot in the dilute to 20 mL with the same solvent.
chromatogram obtained with solution (2). Piau TLC octad«y/silyl silica gel Fm piau R.
Loss on drying Mobile phase 58.4 gIL solution of sodium chloride R,
When dried to constant weight at 60° at a pressure not methanol R, tuetone R (20:30:50 VIVIV).
exceeding 0.7 kPa, loses not more than 1.0% of its weight.
Applicatwn 5 ~L.
Use I g.
Development In an unsaturated tank, over 3/4 of the plate.
Sulfated ash
Drying In air.
Nor more than 0.1 %, Appendix IX A.
ASSAY
System suiUtbiJity Reference solution (b):
Dissolve 0.5 gin 15 mL of ethanol (96%) previously - the chromatogram shows 2 clearly separated spots.
neutralised to phenol red solution and titrate with 0.1M sodium
hydroxide VS using phenol redsolution as indicator. Each mL Results The principal spot in the chromatogram obtained
of 0.1M sodium hydroxide VS is equivalent to 14.82 mg of with the test solution is similar in position and size to the
CgHsOzo
solution (a).
D. Dissolve 0.2 g of anhydrous citric acidR in 10 mL of acetic
anhydride R in a water-bath at 80°C and maintain the
temperature of the water-bath at 80 °C for 10 min.
Cinnarizine Add about 20 mg of the substance to be examined. A purple
(ph. Bur. monograph 0816) colourdevelops.
TESTS
than reference solution BY7 (2.2.2, Method II).
Dissolve 0.5 g in methylene chloride R and dilute to 20 mL
Acidity or alkalln1ty
368.5 298-57-7 Suspend 0.5 g in 15 mL of water R. Boil for 2 min. Cool and
filter. Dilute the filtrate to 20 mL with carbon dioxide-free
Action and use water R. To 10 mL of this solution add 0.1 mL of
Histamine HI receptor antagonist; antihistamine. phenolphthalein solution Rand 0.25 mL of 0.01 M sodium
hydroxide. The solution is pink. To 10 mL of the solution
Preparation
add 0.1 mL of methyl redsolution R and 0.25 mL of 0.01 M
Cinnarizine Tablets
hydrochloric acid. The solution is red.
PhEil _
Related substances
DEFINITION Liquid chromatography (2.2.29).
(E)-I-(D iphenylmethyl)-4-(3-phenylplOp-2-enyl)piperazine. Test solution Dissolve 25.0 mg of the substance to be
Content
99.0 per cent to 101.0 per cent (dried substance). solvent.
Reference solution (a) Dissolve 12.5 mg of cinnarizine CRS
CHARACTERS and 15.0 mg ofjlunarizine dihydrochloride CRS in methanol R
Appearance and dilute to 100.0 mL with the same solvent. Dilute-Lu mL
White or almost white powder. of thissolution to 20.0 mL with methanol R.
Solubility Reference solutwn (b) Dilute 1.0 mL of the test solution to
Practically insoluble in water) freely soluble in methylene 100.0 mL with methanol R. Dilute 5.0 mL of this solution to
chloride, soluble in acetone, slightly soluble in ethanol 20.0 mL with methanol R.
(96 per cent) and in methanol.
Column:
- size: 1== 0.1 m, 0 = 4.0 mm;
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1-586 Ciprofibrate 2022
- srationary phase: base-deactivated octadecylsilyl silica gelfor

chromatQgraphy R (3 urn).
Mobile phase:
- mobile phase A: 10 gIL solution of ammonium aatale R;
- mobile phase B: 0.2 per cent VIV solution of glacial acetic
acidR in acetonim"le Rl,
B. (Z)-I-(diphenylmethyl)-4-(3-phenylprop-2-enyl)piperazine,
Time MobUe phase A MobIle phBIJe B
(mln) {per cent Vm (per cent VIJI)
75 -+ 10 25 ...... 90
10 90
Flaw rate 1.5 mllmin.

Detection Spectrophotometer at 230 run. C. 4-(diphenylmethyl)-l, I-bis[(Ej -3-phenylprop-2-enyl]
pjperazinium chloride,
[nj«,;on I0 ~L.
Relative retention Withreference to cinnarizine (retention
=
time about II min): impurity A about 0.4;=
flunarizine = about 1.05; impurity B = about 1.1;
impurity C = about 1.2; impurity D = about 1.6;
=
- resolution: minimum 5.0 between thepeaks due to
cinnarizine and flunarizine. D.l-(diphenylmethyl)-4-[(IRS,3Ej-4-phenyl-l-[(Ej-2-
phenylethenyl]but-3-enyl]piperazine,
Limits:
- impurities A, H, C, D, E: for each impurity, not more than
- unspecified impurities: foreach impurity, not more than
(0.10 per cent);
- total: not more than twice the area of the principal peak in E. 1,4-bis(dipbeoyhnethyl)piperazine.
the chromatogram obtainedwith reference solution (b) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEur
(0.5 per cent);
- disregard lim,;: 0.2 times the area of the principal peak in
(0.05 per cent). Ciprofibrate ***
Loss on drying (2.2.32) *** ***
Maximum 0.5 per cent, determined on 1.000 g by drying in (Ph. Bur. monograph 2013) ***
an oven in vacuo at 60°C for 4 h.
Sulfuted ash (2.4. 14) I"""Y°XCo,H
Maximwn 0.1 per cent, determined on 1.0 g. CI'A"'V H,C CH, and enantiomer
ASSAY CI H
Dissolve 0.150 g in 50 mL of a mixture of! volume of
anhydrous acetic acid Rand 7 volumes of methylethyl kesone R. 289.2 52214-84-3
Titrate with 0.1 M perchwm acid, using 0.2 mL of
naphtho/benzein solution R as indicator. Acdon and use
I mL of O. I M perchlom acid is equivalentto 18.43 mg Fibrate; lipid-regulating drug.
of C,JI"N2 • PhEur _
STORAGE DEFINITION
Protected from light. 2-[4-[(IRSJ-2,2-Dichlorocyclopropyl)phenoxy]-2-
IMPURITIES methylpropanoic acid.
Specified impun'ties A, B, C, D, E. Content
CHARACTERS
Appearance
White or slighdy yellow, crystalline powder.
Solubility
A. l-(diphenylmethyl)piperazine, Practically insoluble in water, freely soluble in anhydrous
ethanol, soluble in toluene.
mp
About 115 "C.
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2022 Ciprofibrate 1-587
IDENTIFICATION - any other ;mpun"ty: for each impurity, not more than the
Infrared absorption spectrophotometry (2.2.24). area of the principal peak in the chromatogram obtained
Comparison ciprofibrate CRS. with reference solution (a) (0.1 per cent),
- total of other impurities: not more than 5 times the area of
TESTS the principal peak in the chromatogram obtained with
Appearance of solution reference solution (a) (0.5 per cent),
The solution is clear (2.2.1) and not more intensely coloured - disregard limit: 0.5 times the area of the principal peak in
than reference solution BY, (2.2.2, Method If). the chromatogram obtainedwith reference solution (a)
Dissolve 1.0 g in anhydrous ethanol R and dilute to 10.0 mL (0.05 per cent).
with the same solvent. Chlorides (2.4.4)
Related substances Maximum 350 ppm.
Liquid chromatography (2.2.29). To 0.190 g add 20 mL of water R and treat in an ultrasonic
Test solution Dissolve 0.125 g of the substance to be bath for 8 min. Filter. 15 mL of the filtrate complies with the
examined in a mixture of equal volumes of acetonim1e R and test.
water R and dilute to 50 mL with the same mixture of Water (2.5.I'l)
solvents. Maximum 0.5 per cent, determined on 1.000 g.
100.0 mL with a mixture of equal volumes of acetonitrile R
and water R. Dilute 1.0 mL of this solution to 10.0 mL with
a mixture of equal volumes of acetonitrile R and water R. ASSAY
Reference solution (b) Dissolve the contents of a vial of Dissolve 0.250 g in a mixture of 20 mL of water Rand
ciprofibrate for system ,uitability CRS in 2.0 mL of a mixture of 40 mL of anhydrous ethanol R. Titrate with 0.1 M sodium
equal volumes of acesonioile R and water R. hydroxide, determining the end-point potentiometrically
(1.2.10).
Column:
- size: 1= 0.15 ID, 0 == 4.6 mm, 1 mL of 0.1 M sodium hydroxide is equivalent to 28.92 mg
- 'tationaryphase: il<tylsilyl silica gelfor chromatography R of C 13H14CIz03 .
(5 urn). STORAGE
Mob,7e phase: In an airtight container, protected from light.
- mobile phaseA: 1.36 gIL solution of potassium dihydrogen IMPURITIES
phosphxue R adjusted ro pH 2.2 with phosphoric acidR, Specified impuri,Ws A, B, C, D, E.
- mobile phase B: acetonitrile R,

(min) (per cent V/l? (per cent JIm
• - 30 75 ---0 30 25 ---0 70
30 - 40 3. 70
A. 2-(4-ethenylphenoxy)-2-methylpropanoic acid,
40 - 42 30 ---0 75 70 ---0 25
r'1(0H
C1 "A.. V and eneoucmer
Injection 10 ~L. C, H
Identification of impun"u"es Use the chromatogram supplied

B. 4-[(1 R5)-2,2-dichlorocydopropyl)phenol,
with ciprofibrate for system ,"itab~i,y CRS to identify the peaks
due to impurities A, B, C, D and E.
Relative retention With reference to ciprofibrate (retention
=
time about 18 min): impurity A about 0.7; = andenantiomer
=
impurity B about 0.8; impurity C about 0.95; =
impurity D = about 1.3j impurity E = about 1.5.
System suitability Reference solution (b): C. 2- [4- [(1R5) -2,2-dichlorocydopropyl] phenoxy]-2-
- resolution: baseline separation between the peaksdue to methylpropan-l-ol,
impurity C and ciprofibrate.
Limits:
peak area of impurity A by 2.3,
- impurities A, C, D: for each impurity, not more than the
with reference solution (a) (0"1 per cent),
- ;mpm;ly B: not more than twice me area of the principal D. methyl 2-[4-[( IR5)-2,2-dichlorocydopropyl]phenoxy]-2-
merhylpropanoate,
- impun'ty E: not more than 8 times the area of the principal
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1-588 Ciprofloxacin 2022
Plate TLC silica gelFm plate R.

Mobile phase acetonitrile R, concentrated ammonia R,
methanol R, methylene chloride R (lO:20:40:40 VIVIVIV).
Applicatian 5 ~L.
Development At the bottom of a chromatographic tank,
E. ethyl 2-[4-[(IRS)-2,2-dichlorocydopropyl]phenoxy]-2- place an evaporating dish containing 50 mL of concentrated
methylpropanoate. ammonia R. Expose the plate £0 the ammonia vapour for
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE" 15 min in the closed tank. Withdraw the plate, transfer to a
2nd chromatographic tank and develop over 3/4 of the plate.
Drying In air.
Ciprofloxacin Limit:
- itnpun'ty A: any spot due to impurity A is not more
(ph. Eur. monograph 1089) intense than the principal spot in the chromatogram
obtainedwith the reference solution (0.2 per cent).
Related substances
Test solution To 25.0 mg of the substance to be examined
add 0.2 mL of dilute phosphoric acid R and dilute to 50.0 mL
with the mobile phase. Treat in an ultrasonic bath until a
clear solution is obtained.
331.4 85721-11-1 Reference solution (a) Dilute 1.0 mL of the lest solution to
Action and use solution to 5.0 mL with the mobile phase.
Fluoroquinolone antibacterial. Reference solution (b) Dissolve 2.5 mg of ciproflo.wdn
Preparations hydrochloride for peak identijication CRS (containing
Ciprofloxacin Eye Drops lmpunries B, C, D and E) in the mobile phase and dilute to
Ciproftoxacin Infusion 5.0 mL with the mobile phase.
Ciprofloxacin Oral Suspension Go/umn:
- size: / = 0.25 m, 0 = 4.6 mm;
PhE" _ - stationary phase: base-deaawoted end-eapped octadecylsi1yl
DEFINITION silica gelfor chromacagraphy R (5 ~m);
l-Cyclopropyl-t-fluoro-d-oxo-?-(pipernzin-I-yl)-I,4-
dihydroquinoline-3-carboxylic acid. Mobile phase Mix 13 volumes of acetonitrile Rand
87 volumes of a 2.45 gIL solution of phosphoric acid R,
Content
previously adjusted 10 pH 3.0 with triethylamine R.
Flora rale 1.5 mUmin.
CHARACTERS
Dueaion Spectrophotometer at 278 run.
Appearance
Almost white or pale yellow, crystalline powder, slightly Injection 50 IlL.
hygroscopic. Run time Twice the retention time of ciprofloxacin.
Solubility Identification of impurities Use the chromatogram supplied
Practically insoluble in water, veryslightly soluble in with ciprofloxacin hydrochloride for peak idenlijicaU'on CRS and
anhydrous ethanol and in methylene chloride. the chromatogram obtained with reference solution (b) to
identify the peaks due to impurities B, C, D and E.
IDENTIFICATION
Relative retention With reference to ciprofloxacin (retention
= =
time about 9 min): impurity E about 0.4;
Comparison ciprofloxacin CRS. impurity B = about 0.6; impurity C = about 0.7;
TESTS impurity D = about 1.2.
Appearance of solution System suitability Reference solution (b):
The solution is clear (2.2.1) and not more intensely coloured .L resolution: minimum 1.3 between the peaks due to
than reference solution GY, (2.2.2, Method If). impurities Band C.
Dissolve 0.25 g in a 10.3 gIL solution of hydrochlcric acid R Limits:
and dilute to 20 mL with the same solution. - correaion factors: for the calculation of content, multiply
Impurity A the peak areas of the following impurities by the
Thin-layer chromatogmphy (2.2.27). corresponding correction factor: impurity B = 0.7;
impurity C =0.6; impurity D = 1.4; impurity E = 6.7;
Test soluticn Dissolve 50 mg of the substance to be - impurities B, CJ DJ E: for each impurity, not more than
examined in dilute ammonia RJ and dilute to 5 mL with the
same solvent.
Reference solution Dissolve 10 mg of ciprofioxacin - unspeafied impurities: for each impurity, not more than
impurity A CRS in a mixture of 0.1 mL of dilute ammonia Rl 0.5 times the area of the principal peak in the
and 90 mL of water R and dilute [0 100 mL with water R.
Dilute 2 mL of the solution to 10 mL with water R.
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2022 Ciprofloxacin Hydrochloride 1-589

(0.10 per cent);
(0.05 per cent). D. 7-cll1oro-I-cyclopropyl-4-oKo-6-(piperazin-l-yl)-I,4-
Los. on drying (2.2.32) dihydroquinoline-3-carboxylic acid,
Maximum 1.0 per cent, determined on 1.000 g by drying
under vacuum at 120°C.
HNl ?
Sulfated ash (2.4.1 if)
Maximum 0.1 per cent, determined on 1.0 g in a platinum ~Nx:QN
I I
crucible. F "'-
ASSAY o
Dissolve 0.300 gin 80 mL of glacial acetic acid R. Titrate
with 0.1 1\1 perchloric acid, determining the end-point E. l-cyclopropyl-6-f1uoro-7 -(piperazin-I-yl)quinolin-4( 111)-
potentiometrically (2.2.2U). one (decarboxylated compound),
of C17HlSFN303' HNl ?
STORAGE
~N0N
I I
IMPURITffiS HO ' ~H
Specified impurities A, B. C, D, E. o
Otherdetectable impurities (thefollowing subsuznces would, if
F. l-cyclopropyl-6-hydro><y-4-oKo-7-(piperazin-l-yl)-I,4-
dihydroquinoline-3-carboxylic acid.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
criterion for other/unspecified impurities and/or ~ me general
monograph Substances for phannauuti<aJ use (2034). It is
therefore notnecessary to identify these impuriu'e.s for
demonstration ofcompliance. See also 5,10. Control of impurities
Ciprofloxacin Hydrochloride ****
***
in substances for pharmaceutical use) F.
*
(ph. Bur. monograph 0888) *** *
?
C10""
F
I I
"'-
N CO,H
HNl
~N0N
?
o I I
F ' C~H
A, 7-chloro-l-cyclopropyl-6-fluoro-4-oxo-l,4- o
dihydroquinoline-3-carboxylic acid (f1uoroquinolonic
acid), C 17H"ClFN,O,,xH,O 367.8
(anhydrous)
HNl ? Action and use

~NqN
I I
Fluoroquinolone antibacterial.
Preparations
"'- Co,H Ciprofloxacin Ear Drops
o Ciprofloxacin Hydrochloride Eye Drops
Ciproftoxacin Tablets
B. l-cyclopropyl-4-oKo-7-(piperazin-l-yl)-1,4-
dihydroquinoline-3-carboxylic acid (desfluoro compound), PIlE" _
DEFINITION
H ? I-Cyclopropyl-e-fluoro-d-oxo-?-(piperazin-I-yl)-I,4-
dlhydroqulnohne-o-carboxylic acid hydrochloride. It contains
H,N ~N0N
I I a variable quantity of water.
F "'- CO,H Content
o 98.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
C. 7- [(2-aminoethyl)aminoJ-1-cyclopropyl-6-f1uoro-4-oKo- Appearance
1,4-dihydroquinoline-3-carboxylic acid (ethylenediamine Pale yellow, crystalline, slightly hygroscopic powder.
compound),
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1-590 Ciprofloxacin Hydrochloride 2022
Solubility - stationary phase: base-deactivated end-eapped octadecylsilyl

Soluble in water, slightly soluble in methanol, veryslightly silica gelfor chromatography R (5 ~m);
soluble in anhydrous ethanol, practically insoluble in acetone, - temperature: 40 °C.
in ethyl acetate and in methylene chloride. Mobile phase Mix 13 volumes of acetonitrile R and
IDENTIFICATION 87 volumes of a 2.45 gIL solution of phosphoric acidR
A. Infrared absorption spectrophotometry (2.2.24). previously adjusted to pH 3.0 with In'ethylamine R.
Comparison ciprofloxacin hydrochiotide CRS. Flow rate 1.5 mUmin.
B. 0.1 g gives reaction (b) of chlorides (2.3.1). Detection Spectrophotometer at 278 om.
Injection 50 f.l.L of the test solution and reference
TESTS
Solution S
Dissolve 0.5 g in carbon dioxide-free waterR and dilute to Run time 2.3 times the retention time of ciprofloxacin.
20 mL with the same solvent. Identification of impunues Use the chromatogram supplied
with ciprofloxacin hydro<hlOlide for peak identification CRS and
the chromatogram obtained with reference solution (b) to
The solutionis clear (2.2.1) and not more intensely coloured
identify the peaks due to impurities B, C, D and E.
than reference solution GY, (2.2.2, Method 11).
Relativeretention With reference to ciproftoxacin (retention
time e about 9 min): impurity E ;;;; about 0.4;
water R.
impurity B ;;;; about 0.6,; impurity C ;;;; about0.7';
pH (2.2.3) =
Impurity A - resolution: minimum 1.3 between the peaks due to
Thin-layer chromatography (2.2.27). impurities Band C.
Test solution Dissolve 50 mg of the substance to be Limits:
examined in water R and dilute to 5 mL with the same - correction locum: for the calculation of content, multiply
solvent. the peak areas of the following impurities by the
Reference solution Dissolve 10 mg of ciprofloxacin corresponding correction factor: impurity B ;;;; 0.7;
impurity A CRS in a mixture of 0.1 mL of dilute ammonia Rl impurity C = 0.6; impurity D = 1.4; impurity E = 6.7;
and 90 mL of water R and dilute to 100 mL with water R. - impuniy E: not more than 1.5 times the area of the
Dilute 2 mL of the solution to 10 mL with waler R. principal peak in the chromatogram obtained with
Plate TLC silica gel F m 'p/ate R. reference solution (c) (0.3 per cent),
Mobile phase acetonitrile RJ concentrated ammonia R,
- impuruies B, C, D: for each impurity, not more than the
methanol R, me,hykne chloride R (10:20:40:40 VIVIV/Y). area of the principal peak in the chromatogram obtained
Application 5 IJL. - unspecified impurities: for each impurity, not more than
Deoelopment At the bottom of a chromatographic tank, 0.5 times the area of the principal peak in the
place an evaporating dish containing 50 mL of concentrated chromatogram obtained with reference solution (c)
ammonia R. Expose the plate to the ammonia vapour for (0.10 per cent);
15 min in the closed tank. Withdraw the plate, transfer to a - lOtal: not more than 2.5 times the area of the principal
2nd chromatographic tank and develop over 3/4 of the plate. peak in the chromatogram obtained with reference
Drying In air. solution (c) (0.5 per cent);
Detection Examine in ultraviolet light at 254 nm. - disregard limit: 0.25 times the area of the principal peak in
Lim;t:
(0.05 per cent).
- impurity A: any spot due to impurity A is not more
intense than the principal spot in the chromatogram Water (2.5.12)
obtained with the reference solution (0.2 per cent). Maximum 6.7 per cent, determined on 0.200 g.
Liquid chromatography (2.2.29). Maximum 0..1 per cent, determined on 1.0 g in a platinwn
Test sotution Dissolve 25.0 mg of the substance to be crucible.
examined in the mobile phase and dihne to 50.0 mL with ASSAY
the mobile phase. Liquid chromatography (2.2.29) as described in the test for
Reference solution (a) Dissolve 25.0 mg of ciprofloxacin related substanceswith the following modification.
hydrochloride CRS in the mobile phase and dilute to 50.0 mL Injection I0 ~ of the test solutionand reference
with the mobile phase. solution (a).
Reference solu,wn (b) Dissolve 2.5 mg of ciprofloxacin Calculate the percentage content of C17HI9CIFN303 taking
hydrochloride for peak identification CRS (containing into account the assigned content of ciprofloxacin
impurities B, C, D and E) in the mobile phase and dilute to hydrochloride CRS.
STORAGE
Reference solu,wn (c) Dilute 1.0 mL of the test solution to In an airtight container, protected from light.
solution to 10.0 mL with the mobile phase. IMPURITIES
Golumn: Specified impun"ties A, B, C, D, E.
- size: I ;;;; 0.25 m, 0 ;;;; 4.6 mm; Otherdetectable impurities (thefollowing substances would, if
present at a sufficient level, be delated by one or other of the tests
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2022 Cisatracurium Besilate 1-591

Cisatracurium Besilate ***
criterion for other/unspecified impuniies and/or by the general *** ***
monograph Subseances for phannaceutical use (2034). It is (Ph. Eur. monograph 2763) ***
therefore not necessary to identify these ;mpun'ties for
demonstration of compliance. See also5.10. Conaolof impurities
in substances for pharmaceuticaluse) F.
A. 7-chloro-I-cyc1opropyl-6-ftuoro-4-oxo-I,4-
dihydroquinoline-3-carboxylic acid (fluoroquinolonic
acid),
1243 96946-42-8
Action and use

Non-depolarizing neuromuscular blocker.
PIlE" _
B. l-cyc1opropyl-4-oxo-7-(piperazin-I-yl)-I,4-
dihydroquinoline-3-carboxylic acid (desftuoro compound), DEFINITION
2,2 t - [pentane-I,5-diylbis[oxy(3-oxopropane-3,I-diyl)II bis
[(IR,2R)-I-[(3,4-dlmethoxyphenyl)methyl]-6,7-dimethoxy-2-
methyl-l,2 J3,4-tettahydroisoquinolinium] dibenzenesulfonate.
Content
PRODUCTION
It is considered mat alkyl benzenesulfonate esters are
C. 7- [(2-aminoethyl) amino) -I-cyc1opropyl-6-f1uoro-4-oxo-
genotoxic and are potential impurities in cisatracurium
1,4-dihydroquinoline-3-carboxylic acid (ethylenediamine besilate. The manufacturing process should be developed
compound), taking into consideration the principles of quality risk
rN
HN~
c,x:v~
I
y
0
I
C0:2H
management, together with considerations of the quality of
starting materials, processcapability and validation.
The general method 2.5.41. Methyl, ethyland isopropyl
benzenesulfonate in aaiue substances is available to assist
manufacturers,
CHARACTERS
Appearance
D. 7-chloro-l-cyc1opropyl-4-oxo-6-(piperazin-I-yl)-1,4- Whiteor yellowish, hygroscopic powder.
dihydroquinoline-3-carboxylic acid, Solubility
Soluble in water, verysoluble in anhydrous ethanol) freely
HNl Y soluble in methylene chloride.
~Nx:QN
I I
IDENTIFICATION
F "" Comparison ciuuracunum besilate CRS.
o B. Examine the chromatograms obtained in the assay.
E. l-cyc1opropyl-6-ftuoro-7-(piperazin-I-yl)quinolin-4(1H)- Results The principal peakin the chromatogram obtained
one (decarboxyJated compound), with test solution (b) is similar in retention time and size to
TESTS
than reference solution Y7 (2.2.2, ll'1.etJwd II).
F. l-cyc1opropyl-6-hydroxy-4-oxo-7-(piperazin-I-yl)-I,4- same solvent.
dihydroquinoline-3-carboxylic acid.
_~ PIIEII
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1-592 Cisatracurium Besilate 2022
Diastereoisomers - stationary phase: end-capped octadecylsi/y/ silica gelfor

Liquid chromatography (2.2.29): use the normalisation chromatography compatible with 100 per centaqueous mobile
procedure. phases R (2 urn);
Testsolution Dissolve 0.100 g of the substance to be - temperature: 35 "C.
examined in a 0.1 per cent VIV solution of diethykzm;ne R in Mobile phase:
methanol R and dilute to 25.0 mL with the same solution. - mobile phase A: mix 5 volumesof methanol RJ 15 volumes
Heat in a water-bath at 60°C for 15 min then cool to room of acetonitrile Rand 80 volumes of a 6.8 gILsolution of
temperature. potassium dihydrogen phosphate R;
Reference solution Dissolve 8 mg of laudanosine CRS in a - mob.1e phase B: methanol R, acetonitrile R (30:70 Vflo');
0.1 per cent VIV solution of diethylamine R in methanol Rand
dilute to 2.0 mL with the same solution. Tim, Mobile phase A MobUe phase B
(min) (percent Vm (per cent I'm
Column:
0-1 92 -4 82 8 -4 18
- size: 1:::;; 0.25 m, 0 = 4.6 mm;
I -a 82 ---> 80 18 ..... 20
- stationary phase: amylose derivative of silico gelfor chiral
8 - 10 80 ..... 70 20 ..... 30
separation R (5 um).
10 - 12 70 ..... 45 30 ..... 55
Jw'obile phase propanol Rl, 0.1 per cent VIV solution of 12 - 20 45 55
diethylamine R in heptane R (15:85 VflI).
Flrtw rate 0.8 mUmin. Flow rate 0.5 rnUmin.
Detection Spectrophotometer at 285 nm. Detection spectrophotometer at 280 om"
Injection 10 ~L. AUUJSampler Set at 4 "C.
Run time 1.5 times the retention time of R-Iaudanosine. Injection 5 ~L of test solution (a) and reference
Elun"on order S-Iaudanosinej R-Iaudanosine. solutions (b), (c) and (d).
Retention time R-Iaudanosine = about 18 min. Identification of impurities use the chromatogram supplied
System smiabJ7ity Reference solution: with cisatraamum for system suitability CRS and the
- resohuion: minimum 2.0 between the peaks due to chromatogram obtainedwith reference solution (c) [Q identify
S-laudanosine and R-Iaudanosine. the peaks due to impurities D J F J H + T + V~ N~ 0, P, Q;
Calculate the percentage content of the sum of impurities S, use the chromatogram supplied with cisatracutium for peak
T, U and V by multiplying the percentage of S-Iaudanosine identification CRS and the chromatogram obtained with
by 2. reference solution (d) (Q identify the peaks due to impurities
Limit: A. C, 1 (epimer I) and I (epimer 2) + K + L + M.
- sum of impun"ties S, T, U, v: maximum 0.5 per cent; Relativeretention With reference to cisatracurium (retention
disregard any peak other than S~laudanosine and time =about 9 min): impurity A =about 0"3;
R-Iaudanosine" =
impurity C about 0.4; impurity D about 0.45;=
impurity F = about 0.55; impurities H, T
Related substances
= =
and V about 0.97; impurity I (epimer I) about 1.25;
impurities I (epimer 2), K, Land M = about 1.3;
procedure"
impurity N = about 1.4; impurity 0 = about 1.5;
So/vent mixture Mix 20 volumes of acetonitrile Rand
=
impurity P about 1.6; impurity Q about 1.65.=
Impurity M can coelute or be separated from impurities I
phosphate R previously adjusted to pH 3.1 with phospho";"
(epimer 2), K and L.
acid R.
- peak-to-<Jalley ratio: minimum 3, where Hp = height above
the baseline of the peakdue to impurity Hand/or T
and/orV and H; = heightabove the baseline of the lowest
Testsolution (b) Dilute 1.0 mL of test solution (a) to point of the curve separating this peak from the peak due
10.0 mL with the solvent mixture. to cisatracurium.
Reference solution (a) Dissolve 50.0 mg of ciseuracurium Limits:
besilate CRS in the solvent mixture and dilute to 25.0 mL - correction factor: multiply the peak area of impurity C by
with the solvent mixture. Dilute 1.0 mL of the solution to 0.5;
10.0 mL with the solvent mixture. - sum of impun"ries H~ T~ V: maximum 0.8 per cent;
Reference solution (b) Dilute 1.0 mL of lest solution (a) to - impun"ty Q: maximum 0.7 per cent;
100.0 mL with the solvent mixture. Dilute 1.0 mL of this - impurilies A, C~ F, OJ P. for each impurity, maximum
solution to 20.0 mL with the solvent mixture" 0.5 per cent;
Reference solution (c) Dissolve 2.0 mg of cisatracutium for - impun"ty D: maximwn 0.4 per cent;
system sw"tability CRS (containing impurities D, F~ N, OJ P, - impurity N: maximum 0.3 per cent;
Q and Hand/or T and/or V) in 1.0 mL of the solvent - sum of impurities I~ K~ L, 1\1: maximwn 0.3 per cent;
mixture" - unspecified impuruies: for each impurity, maximum
Reference solntum (d) Dissolve 2.0 mg of cisatreuurium for 0.10 per cent;
peak identification CRS (containing impurities AJ C, I (epimer - total: maximum 2.5 per cent;
I) and I (epimer 2) and/or K and/or Land/or M) in 1.0 mL - reporting threshold: 0.05 per cent (reference solution (b»).
of the solvent mixture. Water (2.5.1Z)
Column: Maximum 5.0 per cent, determined on 1.00 g.
- size: 1= 0.10 m, 0 = 3.0 nun;
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Sulfaled ash (2.4.14)

ASSAY
related substances with the following modifications:
Injection Test solution (b) and reference solution (a).
System suitability Reference solution (a): D. (IR,2R)-I-[(3,4-dimethoxyphenyi)methyl]-6,7-dimethoxy-
- symmetry faaor: maximum 2.5 for me principal peak. 2-(3-methoxy-3-oxopropyl)-2-methyl-I,2,3,4-
Calculate the percentage content of C65Hs2N201SS2 taking tetrahydroisoquinolinium,
into account the assigned content of cisatracurium
besilare CRS.
STORAGE
temperature of 2 °C to 8°C.
IMPURITIES
Test for diastereoisomers: S.
Test for related substances: A, B, C, D, H, F, GJ H, I, K,
E. (IR,2S)-I- [(3,4-dimethoxyphenyl)methyl)-2- [3-[(5-
L, M, N, 0, P, ~ R, 1', U V, w.:
J
hydroxypentyl)oxy)-3-oxopropyl]-6,7-dimethoxy-2-methyl-
Specified impurities A, C, D, P, H, I, ~ L, M, N, 0, P, Q, 1,Z,3,4-tetrahydroisoquinolinium,
S, T, V.
Otherde/mabie impurities (thefollowing substances wanld, if
critenon for otherlunspecified impurities andlor fry thegeneral
manograph Substances for phannaceulical use (2034). 11 is
therefore not necessary to idenuJy these impurities for
demonstration 0/compliana. Seealso 5.JO. Control of impurities
in substances for pharmaceutical use) B, E, 0, R, U, W.
F. (IR, 2R)-I-[(3,4-dimethoxyphenyl)methyl)-2- [3-[(5-
hydroxypentyl)oxy)-3-oxopropyl]-6,7-dimethoxy-2-methyl-
l,2,3,4-tetrahydroisoquinolinium,
A. (IR,2R)-2-(2-carboxyethyl)-I-[(3,4-
dimethoxypbenyl)methyl]-6,7-dimethoxy-2-methyl-
I,2,3 ,A-tetrahydroisoquinolinium,
G. 2,2'-[penlane-l,5-diylbis[oxy(3-oxopropane-3,I-diyI)JJbis
B. (IR)-I-[(3,4-dimethoxypbenyl)methyl)-6,7-dimethoxy-2,2- [(IR,2S)-I-[ (3,4-dimethoxyphenyl)methyl]-6,7-
dimethy1-1,2,3,4-tetrahydroisoquinolinium, dimethoxy-2-methyl-l,2,3,4-tetrahydroisoquinolinium),
C. (IR)-I-[(3,4-dimethoxyphenyl)methyl]-6,7-dimethoxy-2-
methyl-I,2,3,4-tetrahydroisoquinoline (lR-laudanosine),
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1-594 Cisatracurium Besilate 2022
H'COx:p
'% I N'~CH,
H:)CO \
H,COyY ~O
~ 0
I 0 0
H,CO H" }-OJO
H3CO ~ . .r:'
I N-CH,
H3CO '
H. (IR,I'R,2R,2'S)-2,2' -[pentane-I,5-diylbis[oxy(3- L. (I R,2R)-I- [(3,4-dimethoxyphenyl)methyl]-2- [3-[3- [[5-[[3-

oxopropane-3, I-diyl) I]bis [1-[(3,4- [(IR,2R)-I-[ (3,4-dimethoxyphenyl)methyl]-6,7-
dimethoxyphenyl)methyl]-6,7-dimethoxy-2-methyl- dimethoxy-2-methyl-3,4-dihydroisoquinolinium-2(1H)-yl)
1,2,3,4-tetrahydroisoquinoliniuml, propanoylloxy]pentyl]oxyl-3-oxopropoxy]-3-oxopropyl]-
6,7-dimelhoxy-2-methyl-l,2 J3,4-tetrahydroisoquinolinium,
I. 2,2'-[[(lRS)-I-methylpentane-I,5-diyl] bis[oxy(3-
oxopropane-3, I-diyl) I]bis [( lR,2R)-I- [(3,4-
dimethoxyphenyl)methyll-6,7-dimethoxy-2-methyl- M.2,2 '- [hexane-I,6-diylbis [oxy(3-oxopropane-3, I-diyl) I]bis
1,2,3,4-tetrahydroisoquinolinium], [(1R,2R)-I- [(3,4-dimethoxyphenyl)methyl]-6,7-
dimethoxy-2-methyl-l ,2,3,4-tetrahydroisoquinolinium],
N. (IR,2S)-I-[(3.4-dimethoxyphenyl)methyll-6,7-dimethoxy-
2-methyl-2-[3- [[5- (prop-2-enoyloxy)pentyll oxy1-3-
oxopropyl]-1,2,3,4-tetrahydroisoq uinclinium,
K. 2,2'-[(3-methylpentane-I,5-diyl)bis[oxy(3-oxopropane-
3,I-diyl)Ilbis [(lR,2R)-I-[(3,4-dimethoxyphenyl)methyl]-
6,7-dimethoxy-2-methyl-l,2,3,4-
tetrahydroisoquinolinium),
O. (IR,2R)-I-[(3,4-dimethoxyphenyl)methyll-6,7-dimethoxy-
2-methyl-2- [3-[[5-(prop-2-enoyloxy)pentylloxy]-3-
oxopropyl]-1,2,3,4-tetrahydroisoquinolinium,
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P. (IR,2SJ -1- [(3,4-dimethoxyphenyl)methyl)-2-[3- [[5- [[3- S. (lR,I'S,2R,2'SJ-2,2'-[pentane-I,5-diylbis[oxy(3-

[( IR)-I-[(3,4-dimethoxyphenyl)methyl]-6,7-dimethoxy- oxopropane-3, I-diyl)]] bis[1- [(3,4-
3,4-dihydroisoquinolin-2(1H)-yl)propanoyl]oxy]pentyl) dimethoxyphenyl)methyl]-6,7-dimethoxy-2-methyl-
oxy]-3-oxopropyl]-6,7-dimethoxy-2-melhyl-I,2,3,4- 1,2,3,4-tetrahydroisoquinoliniuml,
tetrahydroisoquinoliniwn,
T. (lR,1 'S,2R,2' R)-2,2'-[pentane-I,5-diylbis[oxy(3-

Q. (IR,2R)-I-[(3,4-dimethoxyphenyl)methyl]-2-[3-[[5-[[3- oxopropane-3, l-diyl)]]bis[I-[(3,4-
[(I R)-I-[ (3,4-dimethoxyphenyl)methyl]-6,7-dimethoxy- dimethoxyphenyl)melhyl]-6,7-dimelhoxy-2-methyl-
3,4-dihydroisoquinolin-2(1H)-yl)propanoyl]oxy]pentyl] 1,2,3,4-tetrahydroisoquinoliniwn],
oxy]-3-oxopropyl]-6,7-dimethoxy-2-methyl-I,2,3,4-
tettahydroisoquinolinium,
U. (IR,I' S,2S,2'R)-2,2' -[pentane-I,5-diylbis[oxy(3-

oxopropane-3, I-diyt)II bis[1-[(3,4-
R. pentane-I,5-diyl bis[3-[(lR)+[(3,4-dimethoxyphenyl) dimethoxyphenyl)methyl]-6,7-dimethoxy-2-methyl-
methyl]-6,7-dimethoxy-3, 4-dihydroisoquinolin-2(IH)-yl] 1,Z,3,4-tetrahydroisoquinolinium],
propanoate],
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1-596 Cisplatin 2022
IDENTIFICATION
Comparison cisplatin CRS.
Test solution Dilute 1 mL of solution82 (see Tests) {Q
10 mL with dimethylformamide R.
Reference solution Dissolve 10 mg of cisplatin CRS in 5 mL
of dimethylformamide R.
Plate cellulose for chromatography R1 as the coating
substance.
Pretreatment Activate the plate by heating at 150 °C for J h.
V. (IS,I' R,2S,2'SJ-2,2'-[pentane-I,5-diylbis[oxy(3-
oxopropane-3, I-diyl)[jbls [1-[(3,4- Mobile phase acetone R, dimethylformamide R (10:90 VflI).
dimethoxyphenyl)methylj-6,7-dimethoxy-2-methyl- Application 2 ur,
1,2,3,4-tetrahydroisoquinolinium], Development Over 2/3 of the plate.
a Drying In air.
}-CH, Detection Spray with a 50 gIL solution of stannous chloride R
in a mixture of equal volumes of dilute hydrochhm'c acid Rand
water R. Examine after 1 h.
H'CO&
I a ~,--Fa Results The principal spot in the chromatogram obtained
~
HCO.# 0 with the test solution is similar in position, colour and size to
H:CO "'" H.... the principal spot in the chromatogram obtained with the
I N-CH, reference solution.
~co ' C. Add 50 mg to 2 mL of dl1ute sodium hydroxide solution R in
a glass dish. Evaporate to dryness. Dissolve the residue in a
W. (I R,2R) -2-[3-[[5-(acetyloxy)pentyljoxylI-I-[(3,4- mixture of 0.5 mL of nimc acid Rand 1.5 mL of hydrochloric
dimethoxyphenyl)methylj-6,7-dimethoxy-2-methyl-3- acidR. Evaporate to dryness. The residue is orange. Dissolve
oxopropyl-l,2,3,4-tetrahydroisoquinoliniwn. the residue in 0.5 mL of waterR and add 0.5 mL of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PI>E.. ammonium chloride solution R. A yellow, crystalline precipitate
is formed.
TESTS
Solulion SI
Cisplatin ***
*** ***
Dissolve 25 mg in a 9 gILsolution of sodium chloride R in
carbon dioxide-free waterR and dilute to 25 mL with the same
(Ph. Bur. monograph 0599) *** solvent.
Solulion S2
Dissolve 0.20 g in dimethylformamide R and dilute to 10 mL
Appearance of solution 81
PtCI,(NH,h 300.0 1566J-27-1 Solution SI is clear (2.2.1) and not more intensely coloured
than reference solution GY, (2.2.2, Method Ii).
Action and use Appearance of solution 82
Platinum-containing cytotoxic. Solution S2 is clear (2.2.1).
Preparation pH (2.2.1)
Cisplatin Injection 45 to 6.0 for solution SI, measured immediately after
Pl>E.. _ preparation.
Related substances
DEFINITION
Liquid chromatography (2.2.29). Cany out the testprotected
cis-Diamminedich1oroplatinwn(ll). from Ught. Do not heator sonicate Q'tV p/atinum-amtaining
Content solution. AD solutions are to be used within 4 h.
97.0 per cent to 102.0 per cent. Test solution Dissolve 25.0 mg of the substance to be
CHARACfERS examined in 3 9.0 gIL solution of sodium chloride R and dilute
Appearance to 25.0 mL with the same solution.
Yellow powder, or yellow or orange-yellow crystals. Reference solution (a) Dissolve 25.0 mg of c~p1atin CRS in a
Solubility 9.0 gIL solution of sodium chloride R and dilute to 25.0 mL
Slightlysoluble in water, sparingly soluble in with the same solution.
dimethylformamide, practically insoluble in ethanol Reference solution (b) Dissolve 5.0 mg of cisplatin
(96 per cent). impwity A CRS in a 9.0 gIL solution of sodium chloride Rand
Cany out identification test B, the tests (except that for silver) dilute to 50.0 mL with the same solution.
and the assayprotected from light.
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2022 Cisplatin 1-597
Reference solution (c) Dissolve 5.6 mg of cisplatin Test solution Dissolve 0.100 g in 15 mL of nitric acid R)
impurity B CRS in a 9.0 gIL solution of sodium chloride Rand heating to 80 "C. Cool and dilute to 25.0 mL with water R"
dilute to 100.0 mL with the same solution. Reference solutions To suitable volumes (10 mL to 30 mL)
Reference solution (d) Mix 0.05 mL of the test solution with of silver standard solution (5 ppm Ag) R add 50 mL of nitric
5.0 mL of reference solution (b) and 5.0 mL of reference acid R and dilute to 100.0 mL with waterR.
solution (c) and dilute to 25.0 mL with a 9.0 gIL solution of Source Silverhollow-cathode lamp, preferably using a
sodium chloride R. transmission band of 0.5 om.
Reference sdudon (e) Dilute 5.0 mL of reference solution (d) Wavelenth 328 nm.
to 20.0 mL with a 9.0 gIL solution of sodium chloride R.
Atomiuuion device Fuel-lean air-acetylene flame.
Blanh solution 9.0 gIL solution of sodium chloride R.
Carry out a blank determination.
Column:
- size: I;;;; 0.25 m, 0 = 4.0 mm; ASSAY
- stationary phase: base-deactivated octy1sl1yl silica gelfor Liquid chromatography (2.2.29) as described in the test for
chromalography R (4 urn); related substances with the following modification.
- temperature: 30 DC. Injection 10 J.lL of the test solution and reference
J.l1obi/e phase Dissolve 1.08 g of sodium oaanesulfimate R, solution (a).
1.10 g of tetrabutylammonium hydrogen sulfate Rand 2.12 g of Calculate the percentage content of PtC12 (NH3h from the
potassium d;hydrogen phosphate R in waterfor chromatagraphy R sum of the areas of the peaks due to cisplatin and cisplatin
and dilute to 950 mL with the same solvent. Adjust (0 aquo complex and from the declared content of
pH 5.9 with 1 M sodium hydroxide and dilute to 1000 mL cisp/atin CRS.
with waterfor chromatagraphy R. STORAGE
Flow rate 1.0 mUmin. In an airtight container, protected from light"
Detection Spectrophotometer at 210 run. IMPURITIES
Injection 20,..r.. of the test solution, reference solutions (d) Specified imputities A, B.
and (e), and the blank solution.
Other detectable impun",ies (me following substances would, if
Run time 7 times the retention time of cisplatin. present at a sufficient level, be detected by one or other of the tests
The displacement peakis the latesteluting peak of the group in the monograph. They are limited by the general acceptance
of injection peaks in the chromatogram obtained with the cruetion for other/unspecified impurities and/or by thegeneral
blank solution. monograph Substances for phannaceutical use (2034). 1t is
Identification of cisplatin aquocomplex Use the chromatogram therefore not necessary UJ identify these impun'cies for
supplied with cisp/atin CRS and the chromatogram obtained demonstration of compliance. S~ also 5.10. Control of impurities
with reference solution (a) to identify the peak due to in substances for phannaceutical use) c.
cisplatin aquo complex.
Relativeretention With reference to cisplatin (retention
time :::: about 3.8 min): displacement peak :::: about0.5;
impurity A =: about 0.6; impurity B ::::: about 0.7; cisplatin
aquo ccmplex e about 1.2.
A. trans-diamminedichloroplatinum(m (transplatin),
System suitabl7ity Reference solution (d):
impurities A and B, the displacement peak and the peak
due to impurity A are well separated.
Limits:
- impun"ty A: not more than the area of the corresponding
peak in the chromatogram obtained with reference B. anuninetrichloroplatinate(-),
solution (d) (2.0 per cent);
- impurity B: not more than the area of the corresponding 2-
CI" ......Cl
solution (d) (1.0 per cent); [ CI/Pl'CI ]
- unspecified impuruies: for each impurity, not more than

0.5 times the area of the peakdue to cisplatin in the
chromatogram obtained with reference solution (d) C. tetrachloroplarinatelz-).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
(0.10 per cent);
- sum of impurities other than A and B: not more than
2.5 times the area of the peak due to cisplatin in the
chromatogram obtained with reference solution (d)
(0.5 per cent);
- disregard limit: the area of the peak due to cisplatin in the
chromatogram obtainedwith reference solution (e)
(0.05 per cent). Disregard any peak due to the cisplatin
aqua complex.
Silver
Maximum 250 ppm.
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1-598 Citalopram Hydrobromide 2022
- mobile phaseB: dissolve 1.58 g of ammonium formate R in

Citalopram Hydrobromide 500 mL of a mixture of 32 volumes of waterfor
(ph. Eur. monograph 2288) chromatography Rand 68 volumes of acetonitrile for
chromatography R;
F
'1 Time MobUe phase A Mobile phase B
11
and eoeotomer
(min) (per cent VIII) (per cent VIII)
CH • H81 0-2 100 0
?' "'~N"" 3 2 - 25 100 ---> 40 0-060
I 0 CH J 25 - 30 40 60
NC '"
FIuw rate 1.0 mllmin.

CwH"BrFN,O 405.3 59729-32-7
Detection Spectrophotometer at 230 runand, for
Action and use impurity G, at 254 am.
Selective serotonin reuptake inhibitor; antidepressant. Injection 40 ~L.
Preparation Identification of impun'ties Use the chromatogram supplied
Citalopram Tablets with citalopram for system suitability CRS and the
PhE" ---------- identify the peaks due to impurities B, D and G.
DEFINITION Relative retention With reference to citalopram (retention
(IRS)-I- [3- (Dimethylamino)propyl)-I-(4-tluorophenyl)-1,3- =
time about 19 min): impurity G about 0.5; =
dihydroisobenzofuran-5-carbonitrile hydrobromide. =
impurity B about 0.7; impurity D about 0.9. =
Content Syttem suitability Reference solution (b):
99.0 per cent to 101.5 per cent (dried substance). - resolution: minimum 1.5 between the peaks due to
impurity D and ciralopram at 230 am.
CHARACTERS
Limits:
Appearance
- correaion factor. for the calculation of content, multiply the
White or almost white, crystalline powder. peak area of impurity G by 0.6;
Solubility - impun"r;y D: not more than twicethe area of the principal
Sparingly soluble in water and in anhydrous ethanol. peak in the chromatogram obtained with reference
IDENTIFICATION solution (a) (0.2 per cent);
A. Optical rotation (see Tests). - impun·ty B: not more than 1.5 times the area of the
Comparison citalopram hydrobramide CRS. -impun·", Gat 254 nm: not more than 1.5 times the area of
C.lt gives reaction (a) of bromides (2.3.1). the principal peak in the chromatogram obtained with
TESTS reference solution (a) (0.15 per cent);
- Unspecified impun"ties: for each impurity, not more than the
-0.10' to + 0.10'.
Dissolve 1.0 g in methanol R and dilute to 20 mL with the - sum 0/ impuniies other than G: not morethan 5 times the
same solvent. area of the principal peak in the chromatogram obtained
Related substances with reference solution (a) (0.5 per cent);
Liquid chromatography (2.2.29). - disregard limit: 0.5 times the area of the principal peakin
Testsolution Dissolve 50 mg of the substance to be the chromatogram obtained with reference solution (a)
examined in mobile phase A and dilute 100.0 mL with '0 (0.05 per cent).
mobile phase A. Loss on drying (2.2.32)
Reference solution (a) Dilute 1.0 mL of the test solution to Maximum 05 per cent, determined on 1.000 g by drying in
100.0 mL with mobile phase A (solution A). Dilute 1.0 mL an oven at 105°C for 4 h.
of solution A to 10.0 mL with mobile phase A. Sulfated ash (2.4.14)
Reference solution (b). Dissolve the contents of a vial of Maximum 0.1 per cent, determined on l.0 g in a platinum
citalopram for system suitabil1"t.y CRS (containing impurities BJ crucible.
D and G) in 1.0 mL of solution A.
ASSAY
Column: Dissolve 0.300 g in 50 mL of ethanol (96 per unO R and add
- size: 1::::: 0.25 m, 0 = 4.6 mm; 0.5 mL of 0.1 M hydrochloric acid. Carry out a potentiometric
- stationary phase: end-capped octadecylsilyl Sl1ica g,ljor titration (2.2.20), using 0.1 M sodium hydroxide. Read the
chromatography compatible with 100 per centaqueous mobile volume added between the 2 pointsof inflexion.
phases R (4 pm),
C,oH"BrFN,O.
Mobile phase:
- mobile phase A: dissolve 1.58 g of ammonium formate R in IMPURITIES
500 mL of a mixture of 4 volumes of acewnitriie for Specified impurities B, D, G.
chromatography R, 32 volumes of methanol Rl and Other deteaable impm;,ies (thefollowing substances uxndd, if
64 volumes of water for chromatography Rj present at a sufficient level, be detected by one or other 0/ the tests
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2022 Citalopram Hydrochloride 1-599

criterion for other/unspecified impurities andlor by thegeneral
there/ore not neussary to identify these impurities for
in suhstanCiS for phannaceurical use) A, G, E, F.
F F. 3-[(1 RS)-5-bromo-l-(4-f1uorophenyl)-1,3-
dihydroisobenzofuran-I-yl]-N,N-dimethylpropan-I-amine,
andenanOOmar F
"'~N/CH3
o
,
CH,
H,N
"-~N/CH3 and enantiomer
o I
o CH 3
H3 C ' N / " / ' ~./
A. (lRS)-I-[3-(dimethylamino)propyl]-I-(4-f1uorophenyl)- I
],3-dihydroisobenzofuran-5-carboxamide, CH, o
Ci.4-(dimethylamino)-I-[(IRS)-I-[3-(dimethylamino)propyl]-
1-(4-f1uorophenyl)-1,3-dihydroisobenzofuran-5-yl]butan-l-
one.
ns eplrner at c- _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'1>81
and !heIrenanliomers
Citalopram Hydrochloride *****

B. 1-[3-(dimethylamino)propyl]-I-(4-f1uorophenyl)-3- ** **
hydroxy-l,3dihydroisobenzofuran-S-carbonitrile, (Ph. Eur. monograph 2203) ***
~
F
F 'nd eneouomer
. Hel
and enanllorne( "-~N ....CH3
~ I
lOCH,
NC ""
NC
o
360.9 851/8-27-0
C. (3RS)-6-cyano-3-[3-(dimethylamino)propyl]-3-(4- Action and use
f1uorophenyl)isobenzofuran-1 (3H)-one, Selective serotonin reuptake inhibitor; antidepressant.
Preparation
Citalopram Oral Drops
I'I>E" ~ _
DEFINITION
(1RS)-l-[3-(Dimethylamino)propyl]-I-(4-f1uorophenyl)-1,3-
dihydroisobenzofuran-5-carbonitrile hydrochloride.
Content .
D. (IRS)-I-(4-f1uorophenyl)-I-[3-(methylamino)propyl]-1,3- 99.0 per cent to 101.5 per cent (dried substance).
dihydroisobenzofuran-5-carbonitrile,
CHARACfERS
F Appearance
"'~N_CH,
Solubility
, enc enentcmer Very soluble in water, freely soluble in anhydrous ethanol.
)~} I o I
CH, IDENTIFICATION
CI -~ A. Optical rotation (see Tests).
E. 3-[(1 RS)-5-chloro-l-(4-f1uorophenyl)-1,3- Comparison citalopram hydrochlon'de CRS.
dihydroisobenzofuran-I-yl]-N,N-dimethylpropan-I-amine,
TESTS
Solution S
Dissolve 1.0 g in me/hanoi R and dilute to 20 mL with the
same solvent.
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1-600 Citalopram Hydrochloride 2022
Appearance of solution - disregard limit: 0.5 times the area of the principal peak in
Solution SJ examined immediately afterpreparation, is clear the chromatogram obtained with reference solution (a)
(2.2.1) and not more intensely coloured than reference (0.05 per cent).
solution Yo (2.2.2, MethodIf). Loss on drying (2.2.32)
Optical rotation (2.2.7) Maximum 0.5 per cent, determined on 1.000 g by drying in
-0.10° to + 0.10°, determined on solution S. an oven at 105°C for 4 h.
Liquid chromatography (2.2.29). Maximum 0.1 per cent, determined on 1.0 g in a platinum
Test solution Dissolve SO mg of the substance to he crucible.
examined in mobile phase A and dilute to 100.0 mL with ASSAY
mobile phase A. Dissolve 0.250 g in 50 mL of ethanol (96 per cent) R and add
Reference solution (a) Dilute 1.0 mL of me test solution to 0.5 mL of 0.1 M hydrochloric acid. Carry out a potentiometric
100.0 mL with mobile phase A (solution A). Dilute 1.0 mL titration (2.2.20), using 0.1 M sodium hydroxide. Read the
of solution A to 10.0 mL with mobile phase A. volume added between the 2 points of inflexion.
Reference solution (b) Dissolve the contents of a vialof 1 mL of 0.1 M sodium hydroxitk is equivalent to 36.09 mg of
citalopram for sysrem suitability CRS (impurities B and D) in C2oH"ClFN,O.
1.0 mL of solution A.
IMPURITffiS
Column: Spe<ified impurities B.
- size: 1= 0.25 m, 0 -= 4.6 mm;
Otherdete<table impurities (thefollowing substances would, if
- sra,wnary phase: end-capped oaad«ylsilyl silica gelfor
present at a sufficient level, be detected Iry one or otherof the tests
chromatography compatible with 100 per centaqueous mobil.
in themonograph. They are limited by thegeneral acceptance
phases R (4 1lJIl);
- tempera..,,: 40 °C.
Mobile phase: therefore not necessary to identify these impurities for
- mobile phase A: dissolve 1.58 g of ammonium formate R in demonstration 0/compliance. See also 5.10. Control of impurities
500 mL of a mixture of 4 volumes of acetonitrile for in substances for pharmaceutical use) A~ C, D~ E, F.
chromatography R, 32 volwnes of methanol Rl and
64 volumes of waterfor chromatography R; F
- mobile phase B: dissolve 1.58 g of ammonium formate R in
500 mL of a mixture of 32 volumes of water for
chromawgraphy Rand 68 volumes of acetonitrile for andenanUomer
chromatography Rj
H,N
(min) (per cent V/J? (pel' cent VIII) o
0-2 100 0
2 - 25 100 ---> 40 0--->60 A. (IRSJ-I-[3-(dimethylamino)propyl]-I-(4-f1uorophenyl)-
25 - 30 40 60 1,3-dihydroisobenzofuran-5-carboxamide,

Injection 40 ur, itsepimer at c'
and theirenanUomers
with citalopram for system suitabilizy CRS and the
identify the peaks due to impurities Band D.
Relative retention With reference to citalopram (retention B. 1-[3-(dimethylamino)propyl)-I-(4-f1uorophenyl)-3-
time = about 19 min): impurity B = about 0.7j hydroxy-I,3-dihydroisobenzofuran-5-carbonitrile,
System suitability Reference solution (b): F
impurity D and citalopram.
Limits: andenanllorner
- impurity B: not more than 1.5 times the area of the
NC
reference solution (a) (0.15 per cent); o
- unspecified impurities: for each impurity, not more thanthe
area of the principal peak in the chromatogram obtained C. (3RSJ-6<yano-3-[3-(dimethylamino)propyl]-3-(4-
with reference solution (a) (0.10 per cent); f1uorophenyl)isobenzofuran-l (3H)-one,
(0.2 per cent);
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2022 Citric Acid 1-601

Preparation Dry the substance to be examined and the
reference substance at lOS ± 2 °C for 2 h.
andenantiomer
Comparison anhydrous citric acid CRS.
OC. Add about 5 mg to a mixture of I mL of acetic
anhydride Rand 3 mL of pyridine R. A red colour develops.
D. (I RS)-l-(4-11uorophenyl)-I-[3-(methylamino)propyl)-1,3-
D. Dissolve 0.5 g in 5 mL of water R, neutralise using 1 M
dihydroisobenzofuran-5-carbonitrile, sodium hydroxide (about 7 mL), add 10 mL of calcium chloride
solution R and heat to boiling. A white precipitate is formed.
E. Water (see Tests).O
TESTS
and enantiomer
The solution is clear (2.2.1) and colourless or not more
intensely coloured than reference solution Y1, BY? or GY1
(2.2.2, Method II).
E. 3-[(IRS)-5-chloro-l-(4-11uorophenyl)-1,3- solvent.
dihydroisobenzofuran-I-yl)-N,N-dimethylpropan-I-amine,
Readily carbonlsable substances
To 1.0 g in a cleaned test tube add 10 mL of sulfuric acid R
and immediately heat the mixture in a water-bath at
90 ± I °C for 60 min. Cool rapidly immedistely afterwards.
The solution is not more intensely coloured than a mixture
of 1 mL of red primary solutionand 9 mL of yellow primary
solution (2.2.2, Method I).
Oxalic acid
Maximum 360 ppm, calculated as anhydrous oxalic acid.
F. 3-[(IRS)-5-bromo-l-(4-11uorophenyl)-1,3-
dihydroisobenzofuran-I-yl)-N,N-dimethylpropan-I-amine. Dissolve 0.80 g in 4 mL of water R. Add 3 mL of hydrochloric
_ _ _ _ _ _ _ _-,- I'IIEII acidR and 1 g of zinc R in granules. Boil for 1 min. Allow to
stand for 2 min. Transfer the supernatant to a test-tube
containing 0.25 mL ofa 10 gIL solution of phenylhydrazine
hydrochloride R and heat to boiling. Cool rapidly, transfer to a
graduated cylinder and add an equal volume of hydrochloric
Citric Acid 1 ***
** ** acidR and 0.25 mL of a 50 gIL solution of potassium
ferricyanide R. Shake and allow to stand for 30 min. Any pink
Anhydrous Citric Acid *****
colour in the solution is not more intensethan that in a
(ph. Eur. monograph 0455) standard prepared at the same time in the same manner
using 4 mL of a 0.1 gIL solution of oxalic acid R.
Sullate. (2.4.13)
Maximum 150 ppm.
CoB.O, 192.1 77-92-9 Dissolve 2.0 g in distilled water R and dilute to 30 mL with
I'IIEII _ the same solvent.
Aluminium (2.4.17)
DEFINITION
Maximum 0.2 ppm, if intended for use in the manufacture of
2-Hydroxypropane-t,2,3-tricarboxyIic acid.
dialysis solutions.
Content Prescribed solution Dissolve 20 g in 100 mL of water Rand
99.5 per cent to 100.5 per cent (anhydrous substance). add 10 mL of aceta" buffer solution pH 6.0 R.
+CHARACTERS Reference solution Mix 2 mL of aluminium standardsolution
Appearance (2 ppm AQ R, 10 mL of aceta" blf!fer solution pH 6.0 Rand
White or almost white, crystalline powder, colourless crystals 98 mL of woterR.
or granules. Blonk solution Mix 10 mL of aceta" buffer solution pH 6.0 R
Solubility and 100 mL of waterR.
Very soluble in water, freely soluble in ethanol (96 per cent). Water (2.5.12)
mp Maximum 1.0 per cent, determined on 2.000 g.
About 153°C, with decomposition.• Sulfated ash (2.4.14)
IDENTIFICATION Maximum 0.1 per cent, determined on 1.0 g.
First ickmijication: B,OE. +Bacterial endotoxin. (2.6.14)
Second identification: A, C, D, E. Less than 0.5 IV/mg, if intended for use in the manufacture
A. Dissolve 1 gin 10 mL of water R. The solution is strongly of parenteral preparations without a further appropriate
acidic (2.2.4).0 procedure for the removal of bacterial endotoxins.•
I This monograph hasundergone phamuuopoeial hannonisaeion.

Seechapter 5.8 Pharmacopoeial harmonisation:
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1-602 Citric Acid 2022
ASSAY 90 ± 1 °C for 60 min. Cool rapidly immediately afterwards.

Dissolve 0.550 g in 50 mL of wale/" R. Titrate with 1 M The solution is not more intensely coloured than a mixture
sodium hydroxide, using 0.5 mL of phenolphthalein solution R as of 1 mL of red primary solution and 9 mL of yellow primary
indicator. solution (2.2.2, Me/hod/).
I mL of 1 M sodium hydroxide is equivalent to 64.03 mg Oxalic acid
ofC.HaO,. Maximum 360 ppm, calculated as anhydrous oxalic acid.
tLABELLING Dissolve 0.80 g in 4 mL of wale/" R. Add 3 mL of hydrochlaric
The label states, where applicable, that the substance is acid Rand 1 g of zinc R in granules. Boil for 1 min. Allow to
intended for use in me manufacture of dialysis solutions.• stand for 2 min. Transfer me supernatant to a test-tube
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" containing 0.25 mL of a 10 gIL solution of ph"!Ylhydrazine
hydrochloride R and heat to boiling. Cool rapidly, transfer to a
graduated cylinder and add an equal volume of hydrochloric
acidRand 0.25 mL of a 50 gIL solution of potassium
/erricyanide R. Shake and allow to stand for 30 min. Any pink
Citric Acid Monohydrate 1 colour in the solution is not more intense than mat in a
standard prepared at the same time in the same manner
using 4 mL of a 0.1 gIL solution of oxalic acid R.
Sulfates (2.4.13)
Maximum 150 ppm.
Dissolve 2.0 g in distilled waterR and dilute to 30 mL with
CJfgO,,H,O 210.1 5949-29-1 the same solvent.
PIlE" _ A1urnlnJum (2.4.17)
Maximum 0.2 ppm, if intended for use in the manufacture of
DEFINITION
dialysis solutions.
2-Hydroxypropane-I,2,3-tricarboxylic acid monohydrate.
Prescribed solution Dissolve 20 g in 100 mL of water Rand
Content add 10 mL of acetate buffer solution pH 6.0 R.
Reference solution Mix 2 mL of aluminium sumdard solution
tCHARACTERS (2 ppm AQ R, 10 mL of acetate buffer solution pH 6.0 Rand
Appearance 98 mL of wale/" R.
White or almost white, crystalline powder, colourless crystals Blank sohnion Mix 10 mL of acetate buffersolution pH 6.0 R
or granules, efflorescent. and 100 mL of wale/" R.
Solubility Water (2.5.12)
Very soluble in water, freely soluble in ethanol (96 per cent). 7.5 per cent to 9.0 per cent, determined on 0.500 g.
t
IDENTIFICATION Maximum 0.1 per cent, determined on 1.0 g.
Firs' identification: B,OE. tBacteriai endotoxins (2.6.14)
Second identification: A, C, D, E. Less than 0.5 ill/mg, if intended for use in the manufacture
A. Dissolve 1 g in 10 mL of water R. The solution is strongly of parenteral preparations without a further appropriate
acidic (2.2.4).0 procedure for me removal of bacterial endotoxins.•
B. Infrared absorption spectrophotometry (2.2.24). ASSAY
Preparation Dry the substance to be examined and the Dissolve 0.550 gin 50 mL of wale/" R. Titrate with 1 M
reference substance at 105 ± 2°C for 2 h. sodium hydroxide, using 0.5 mL of phenolphthalein sohuion R as
Comparison citn'c acid monohydrate CRS. indicator.
OC. Add about 5 mg to a mixture of 1 mL of acetic I mL of 1 M sodium hydroxide is equivalent to 64.03 mg
anhydride Rand 3 mL of pyridine R. A red colour develops. ofC.HaO,.
D. Dissolve 0.5 g in 5 mL of water R, neutralise using 1 M tSTORAGE.
sodium hydroxide (about 7 mL), add 10 mL of calcium chloride In an airtight container.
solution R and heat to boiling. A white precipitate is fanned.
LABELLING
E. Water (see Tests).O The label states, where applicable, that the substance is
TESTS intended for use in the manufacture of dialysis solutions.•
Appearance of solution _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE"
The solution is clear (2.2.1) and colourless or not more
intensely coloured than reference solution Y1, BY7 or GY7
(2.2.2, Me/hod I/).
Dissolve 2.0 g in wa,ter R and diluteto 10 mL with the same
solvent.
Readily carbonisable substances
To 1.0 g in a cleaned test tube add 10 mL of suljuric acidR
and immediately heat the mixture in a water-bath at
l This m()1/()graph has undergone pharmaropoeial harmonisation,

Seechapter 5.8 Pharmacopoeial harmonisation.
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2022 Cladribine 1-603
Reference solution (b) Dissolve 10.0 mg of 2-droxy-o-n'bose R

Cladribine (impurity E) in dimethylformamide R and dilute to 5.0 mL
(ph. Eur. monograph 2174) with me same solvent. Mix 9 volumes of this solution with
1 volume of me test solution.
Plate TI£ stlica gelFm plateR.
Mobilephase concentrated ammonia R, ethanol (96 per unO R,
ethyl acetate R (20:40:40 VIVIV).
Application 5 ~L as bands of 10 mm; thoroughly dry the
points of application in a current of warm air.
Deuelcpment Over 2/3 of the plate.
Drying In air, then heat at 45 °C for 10 min.
Detection Spray with a solution containing0.5 g of thymol R
in a mixture of 5 mL of sulfuric acid Rand 95 mL of ethanol
285.7 4291-63-8 (96 per cent) R; heat at J10 °C for 20 min or until the spots
appear.
Action and use System suitability Referencesolution (b):
Purine analogue; cytostatic. - the chromatogram shows 2 clearly separated spots.
PhE" ~
Limit:
- impun"IY E: any spot due to impurity E is not more intense
DEFINlTION than the spot in the chromatogram obtained with
2-CWoro-9-(2-deoxy-ll-n-eryll",>-pentofuranosyl)-9H-purin-6- reference solution (a) (0.3 per cent).
amine.
Related substances
Content Liquid chromatography (2.2.29).
Solvent mixture acewnim7e R, waterR (10:90 VIV).
CHARACTERS Testsolution (aJ Dissolve 25.0 mg of the substance to be
Appearance examined in the solvent mixture and dilute to 5.0 mL with
White or almost white, crystalline powder. the solvent mixture.
Solubility Testsolution (bJ Dissolve 20.0 mg of the substance to be
Slightly soluble in water, soluble in dimethyl sulfoxide, examined in the solvent mixture and dilute to 100.0 mL with
slightly soluble in methanol, practically insoluble in the solvent mixture.
acetonitrile.
Reference solution (a) Dissolve 20.0 mg of dadribine CRS in
It shows polymorphism (5.9). the solvent mixture and dilute to 100.0 mL with the solvent
IDENTIFICATION mixture.
A. Specific optical rotation (see Tests). Reference solution (b) Dilute 1.0 mL oftest solution (a) to
B. Infrared absorption spectrophotometry (2.2.24). 100.0 mL with the solvent mixture.
Comparison dadribine CRS. Reference solution (c) Dilute 1.0 mL of reference solution (b)
If the spectra obtained in the solid stateshow differences,
dissolve the substance to be examined in the minimum Reference solution (d) Dissolve 1.0 mg of dadribine
volwne of melhanol R and evaporate to dryness. DIY the impurityC CRS in reference solution (b) and dilute to
precipitate at 100°C for 2 h and record a new spectrum 25.0 mL with the same solution.
using the residue. Reference solution (e) Dilute 5.0 mL of reference solution (c)
TESTS
Appearance of solution Reference solution (j) Dissolve 3 mg of cladribine for peak
The solution is clear (2.2.1) and colourless (2.2.2, identification CRS (containing impurities A, B, C and D) in
MethodIi). 2 mL of the solvent mixture.
Disperse 0.15 g in water R, dilute to 50 mL with the same Column:
solvent and sonicate until dissolution is complete. - size: I;;;; 0.25 m, 0 = 4.6 mm;
- stationary phase: base-deactivated oclylsi/yl silica gelfor
Specific optical rotation (2.2.7) chromarography R (5 pm).
-21'.0 to -27.0 (anhydrous substance).
Mobile phase:
Dissolve 0.25 g in dimethyl sulfoxide R and dilute to 25.0 mL - mobile phaseA: waterfor chromatagraphy R;
with the same solvent. - mobile phase B: acetonitrile for chromatography R;
ImpurityE - mobile phase C: 50 gIL solution of phosphoric acid R in
Thin-layer chromatography (2.2.27). waterfor chromatography R;
examined in dimethylfONnamide R and dilute to 2.0 mL with Time Mobile phase A MobUe phase B Mobile phase C
(min) (per cent VIV) (per cent VIV) (per cent V/J1
the same solvent.
0-10 80 -+ 70 10 -+ 20 10
Reference solution (a) Dissolve 5.0 mg of 2-droxy-o-ribose R
10 ~ 25 70 -+ 20 20 -+ 70 10
(impurity E) in dimethy/formamide R and dilute to 25.0 mL
25 - 30 20 70 10
with the same solvent. Dilute 3.0 mL of this solution to
10.0 mL with dimethylformamide R.
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1-604 Cladribine 2022
Detection Spectrophotometer at 252 run. demonstration of compliance. See also 5.10. Control of impurities
Injection 20 ilL of test solution (a) and reference in substances for pharmaceutical use) F, G.
solutions (c), (d), (e) and (I).
:x >
NH
Identification of impundes Use the chromatogram supplied ' N
with clodribine for peak idemlficaiion CRS and the
N I
~0
chromatogram obtained with reference solution (1) to identify
the peaks due to impurities A, B, C and D.
Relativeretention With reference (Q cladribine (retention
=
impurity D about 0.92. OH
- resolution: minimum 4.5 between me peaks due to A. 9-(2-deoxy-p-o-erythm-pentofuranosyl)-9H-purin-2,6-
impurity C and c1adribine. diamine,
Limits:
- correction factors: for the calculation of content) multiply
the peak areas of me following impurities by the
=
corresponding correction factor: impurity B 1.7,;
=
impurity C 0.8;
- impumies A, C: for each impurity, not more than 3 times
obtained with reference solution (c) (0.3 per cent);
- impun'ties B, D: for each impurity, not more than twice
obtained with reference solution (c) (0.2 per cent); B. 9-(2-deoxy-p-o-erythm-pentofuranosyl)-2-methoxy-9H-
- unspecified impurities: for each impurity, not more than the purin-ti-amine,
chromatogram obtained with reference solution (e) C. 2-chloro-7H-purin-6-amine (2-chloroadenine),
(0.05 per cent).
HO~X;NYCl
Water (2.5.32)
OH ( N
Less than 3 IU/mg, if intended for use in the manufacture of N A
procedure for the removal of bacterial endotoxins. Nfl,
ASSAY D. 2-chloro-9-(2-deoxy-<t-D-erythm-pentofuranosyl)-9H-purin-
Liquid chromatography (2.2.29) as described in the test for ri-amine,
lnj"tion Test solution (b) and reference solution (a).
HO\---O~
Calculate the percentage content of ClOH12CINS03 from the
\--/ OH
declared content of c1adn"bine CRS.
STORAGE OH
Protected from light, at a temperature of 2 °C to 8 °C. If the
substance is sterile, store in a sterile, airtight, tamper-evident E. 2-deoxy-o-erythro-pentofuranose (z-deoxy-n-ribose),
container.
LABELLING
preparations.
IMPURITIES F. 4-methylbenzamide,
Specified impun'oo A, B, C, D, E.
cruetion for other/unspeajied impurities andlor by the general
monograph Substances for pharmaceutical use (2034). I, is
G. methyl 4-methylbenzoate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEt6
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2022 Clarithromycin 1-605
Reference solurion (a) Dissolve 75.0 mg of

Clarithromycin claruhromycin CRS in 25 mL of acetonitrile RJ and dilute to
(Ph. Eur: monograph J65J) 50.0 mL with waler R.
o to 100.0 mL with a mixture of equal volumes of
H,C
acetonitrile RJ and water R.
H--
H,C Reference solution (c) Dilute 1.0 mL of reference solution (b)
CH3 H3CO-- to 10.0 mL with a mixture of equalvolumes of acetonitrile RJ
1- oo----!--.. and water R.
H3C03 'j Ho~H~ Reference solution (d) Dissolve 3.0 mg of daiithromycin for
'L-...(
OH
CH3
OCH 3
.{
H
peak identification CRS in 1.0 mL of tuetonitrile RJ and dilute
Blank sohuion Dilute 25.0 mL of acetonitrile RJ to 50.0 mL
CH, with water R and mix.
Column:
748 8JJ03-lJ-9 - size: I;;: 0.10 m, 0 ;;: 4.6 mm;
- stationary phase: octadecylsi/yl silica gelfor chromatography R
Action and use (3.5 um);
Macrolide antibacterial. - temperature: 40 "C.
Preparations Mobile phase:
Clarithromycin Granules for Oral Suspension - mobile phase A: a 4.76 gIL solution of patassium dihydrogen
Clarirhromycin for Infusion phosphare R adjusted to pH 4.4 with dilure phosphoric
Clarithromycin Tablets acid R or a 45 gIL solution of pomssium hydroxide R,
filtered through a CI8 filtration kit;
Clarithromycin Prolonged-release Tablets
- mob-ile phaseB: aceronitriJe Rl,
PhE" _
DEFINITION (min) (per cent V/l? (per cent II/l?
(3R,4S,5S,6R, 7R,9R,IIR,12R, 13S, 14R)-4-[(2,6-Dideoxy-3- 0-32 75 _ 40 25 ...... 60
C-methyl-3-O-methyl-«-L-rib<>-hexopyranosyl)oxy]-I4-ethyl- 32·34 40 60
12, 13-dihydroxy-7-metl-ioxy-3,5,7,9,11,13-hexamethyl-6-
[[3,4,6-trideoxy-3-(dimethylamino)-P-D-.ryI<>-hexopyranosyl)
oxy]oxacyclotetradecane-2,IO-dione (6-0-
methylerythromycin A). Detection Spectrophotometer at 205 om.
Semi-synthetic product derived from a fermentation product. Injection 10 f.lL of the blanksolution, the test solution and
reference solutions (b), (c) and (d).
Content
96.0 per cent to 102.0 per cent (anhydrous substance). Relative retention r (not rG) with reference to c1arithromycin
(retention time ;;: about 11 min): impurity I ;;: about 0.38;
CHARACTERS impurity A = about 0.42; impurity J = about 0.63;
Appearance impurity L = about 0.74; impurity B = about 0.79;
White or almost while) crystalline powder. impurity M = =
about 0.81; impurity C about 0.89;
Solubility impurity D = about 0.96; impurity N = about 1.15;
PracticaUy insoluble in water) soluble in acetone and in impurity E =about 1.27; impurity F =about 1.33;
methylenechloride, slightly soluble in methanol. impurity P = about 1.35; impurity 0 = about 1.41;
impurity K =about 1.59; impurity G =about 1.72;
IDENTIFICATION
impurity H = about 1.82.
System sur·lability:
Comparison: clarithromycin CRS. - symmetry factor. maximum 1.7 for the peak due to
TESTS clarithrcmycin in the chromatogram obtainedwith
Solution S reference solution (b);
Dissolve 0.500 g in methylene chloride R and dilute to - peak-to-ualley ratio: minimum 3.0, where Hp = height
50.0 mL with the same solvent. above the baselineof the peak due to impurity D and
H v ;;: height above the baselineof the lowest point of the
Solution S is clear or not more opalescent than reference CUlVe separating this peak from the peak due to
suspension II (2.2.1) and not more intensely coloured than c1arithromycin in the chromatogram obtained with
reference solution Y7 (1.2.2, Method11). reference solution (d).
Limits:
- correction factors: for the calculation of contents) multiply
-102 to -94 (anhydrous substance), determined on
solution S.
corresponding correction factor: impurity G ;;: 0.27;
Related substances impurity H ::;; 0.15; use the chromatogram supplied with
Liquid chromatography (2.2.29). clarithromycin for peak identification CRS to identify the
Test solution Dissolve 75.0 mg of the substance to be peaks;
examined in 25 mL of acetomttile RJ and dilute to 50.0 mL - any impmity: not more than twice the area of the principal
with water R. peak in the chromatogram obtained with reference
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1-606 Clarithromycin 2022
solution (c) (1.0 per cent), and not more man 4 such o
H,C
peaks have an area greater than 0.8 times the area of the H-···
principal peak in the chromatogram obtained with H,C
reference solution (c) (0.4 per cent); CH3 HJCO'
- wtal: not more than 7 times the area of the principal peak } -O O ,
~NH Ho~Ho
H'C<r..) ,
(3.5 per cent); CH3 Hi\O
- disregard limi,: 0.2 times the area of the principal peak in OH OCH:J H CH3
(0.1 per cent); disregard the peaks eluting before CH,
impurity I and after impurity H.
Water (2.5.12) D. 3"-N-demethyl-6-()..methylerythromycin A,
o
Sulfated ash (2.4.14) H,C
Maximwn 0.2 per cent, determined on 0.5 g. H····
H,C
ASSAY CHJ H3CO
hoo ,
related substances with the following modifications. H,C<qN
...
CHJ
H'
Injection Test solution and reference solution (3). HO~O'
CH3 H 0
Calculate the percentage content of C3sH69N013 taking into OH OCHJ H CH3
account the assigned content of dan·thromydn CRS.
IMPURITlliS CH,
Specified impurities A, B, C, D, E, F, GJ H J 1, J, X, L, M,
N,O,P. B. 6,1l-di-()..methylerythromycin A,
o o
H,C H,C .CHJ
H-·_· H··· ; H
H,C H,C pCH J
CH3 H3CO··· CH3 H 3CO -··· CH,
L -0 0-'-----,;'-.., ~OO , ····H
H3C~3)J H' H'c<q~3 H' CH,
"--{ HO~H,
0 0 Hi '. HO~O
CH Hl\O
3
OH OCH3 H i OH OCH3 H CHJ
HO
CH, CH,
A. 2-<1emethyl-2-(hydroxymethyl)-6-()..methylerythromycin A F. 6,12-di-()..methylerythromycin A,
(clarithromycin F)J
o H,CO'N
H,C H,C
H···· H···
H,C H,C
CH, H,CO ... CH3 H3CO ' "
I 00 ; ho o ,
H'c<q~3 H·
CH3
H,C<qN
... H'
HO~O
CH3 H'.. HO~O
CH3 HI
OH OCH3 . OH OCHJ H
CH, CH,
B. 6-()..methyl-15-norerythromycin A, G. e-Ccmethylerythromycin A (E)-9-«)..methyloxime),

HO.... o
N
H,C H,C
H··· H ....
H,C H,C
CH3 HJCO··· CH:J H3CO····
I 00 . hoo---f..-..
H'c<q~J OHC(~ ...... CHJ ') HI
yH~01Hi
HI
HO~O'
OH
CH H/\O
J
OCH3 . H CHJ OH ? ".
H CH 3
0
CH, CH,
C. ri-Cxmethylerythromycin A (E)-9-oxime, H. 3"-N-demethyl-3'-N-formyl-6-0-methylerythromycin A,
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2022 Clebopride Malate 1-607
o o
CH, H,C
: H H--
.oH H,C
• CH3 CH3 H3CO--
"H ~oo
CH3
.
CH, H,CfQN/ H'
OH
HO~O
CH3
OCH3
H:
H CH3
0
I. 3-0-decladinosyl-6-0-methylerythromycin A, CH,
HO'N N. (10E}-IO,II-didehydro-ll-deoxy-6-0-
H,C
methylerythromycin A,
H
H,C
OCH3
CH3 HO···· N ....
I oo---f-... H,C CH,
H3C~3"j H/ H··· .. H
'L-( H~01H/
H,C OH
OH?
CHl H3CO· ..· CH,
I 00 . ····H
H'CfQ~3 Hi CH,
CH,
HO~O'
CH3 H"\ °
OH OCH3 H CH 3
J. erythromycin A (E)-9-oxime,
CH, CH,
O. 6-0-methylerythromycin A (Z)-9-(O-methyloxime),
o
H,C CH,
H···· i H
H,C OH
CH3 H3CO'" CH,
~OO
CH
• ····H
K. (IS,2R,5R,6S, 7S,8R,9R,11 Z)-2-ethyl-6-hydroxy-9- H,C
..... f Q
3 H' CH,
methoxy-I,5,7,9, II, 13-hexamethyl-8-[[3,4,6-tlideoxy-3- N CH30~0
CH3
•
H'\O
(dimethylamino)-~-D-xyh>-hexopyranosylloxyJ-3,15 OH OCH3 H CH3
dioxabicyclo[l 0.2.1 ]pentadeca-II,13-dien-4-one (3-D-
decladinosyl-8,9:IO,II-dianhydro-6-D- CH,
methylerythromycin A-9,12-hemiketal),
OH P. 4',6-di-D-methylerythromycin A.
N ....
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
H,C ,CH3
H··· : H
H,C OH
CH3 ~co···· / CH3
~oo / "'H
Clebopride Malate
..... C~
H,CfQN H CH,
HO~O'
CH3 H/"" O (ph. Eur. monograph 1303)
OH ocs, H CH3
- 0 (~~ HOXCO,H
Cl~)...J V
CH,
L. 6-D-methylerythromycin A (Z)-9-oxime, AJl~~

~N ~ OCH3
. H> HO,C
and eoantlomer
HO'N
H,C CH, 508.0 57645-91-7
H···· i H
H,C OH Acdon and use
CH3 H3CO" CH, Dopamine receptor antagonist; antiprotozoal (veterinary).
H'C~)---O0 I
··H
CH, PIlE" _
~NH Ho~Ho
CH
3 H/\O DEFINITION
OH OCH3 H CH3 4-Amioo-N- (l-benxylpiperidin-4-yl)-5-chloro-2-
methoxybenzamide acid (RS}-2-hydroxybutanedio.te.
CH,
Content
M.3"-N-demethyl-6-0-methylerythromycin A (E}-9-oxime,
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1-608 Clebopride Malate 2022

Appearance Liquid chromatography (2.2.29).
White or almost white, crystalline powder. Test solution Dissolve 0.100 g of the substance to be
Solubility examined in the mobile phase and dilute to 100.0 mL with
Sparingly soluble in water and in methanol) slightly soluble in the mobile phase.
anhydrous ethanol, practically insoluble in methylene Reference souuion (a) Dilute 1.0 mL of the test solution to
chloride. 100.0 mL with the mobile phase. Dilute 1.0 mL of this
mp solution to 10.0 mL with the mobile phase.
About 164 °C, with decomposition. Reference solution (b) Dissolve 10 mg of the substance to be
examined and 10 mg of metodopromide hydrochloride CRS in
IDENTIFICATION
Second identification: A, C, D. mobile phase.
(2.2.25). - size: 1= 0.12 m, 0 = 4.0 mm;
Test solution Dissolve 20.0 mg in waterR and dilute to - stationary phase: oCladecylsilyl silica gelfor chromatography R
100.0 mL with the same solvent. Dilute 10.0 mL of the (5 urn).
solution to 100.0 mL with water R. Ivfobile phase Mix 20 volwnes of acetonitrile RJ and
Spearal range 230-350 om. 80 volumes of a 1 gIL solution of sodium heptanesulfonate R
Absorption maxima At 270 nm and 307 run. adjusted to pH 2.5 with phosphori< acidR.
Specific absorbance at the absorption maxima: FloW rate 1 mllmin.
- at 270 om: 252 to 278; Detection Spectrophotometer at 215 nm.
- at 307 om: 204 to 226. Inje<tion 20 ~L.
B. Infrared absorption spectrophotometry (2.2.24). Run time Twice the retention time of c1ebopride.
Comparison ckbopride malate CRS. Relative retention With reference to clebopride (retention
C. Dissolve 20 mg in I mL of sulfunc acid R, add I mL of =
time about 15 min): metoclopramide = about 0.45.
p-naphthol solution Rl and mix. The solution examined in System suitability Reference solution (b):
daylight is yellow with blue fluorescence. - resolution: minimum 5.0 berween the peaks due to
D. Thin-layer chromatography (2.2.27). metcclopramide and clebopride.
Test so/mum Dissolve 5 mg of the substance to be examined Limits:
in anhydrous ethanol R and dilute to 10 mL with the same - unspecified impurities: for each impurity, not more than the
solvent. area of the principal peak in the chromatogram obtained
Reference solution (a) Dissolve 5 mg of debopnde malare CRS with reference solution (a) (0.10 per cent);
in anhydrous ethanolR and dilute to 10 mL with the same - total: not more than 3 times the area of the principal peak
solvent. in the chromatogram obtained with reference solution (a)
Reference solution (b) Dissolve 5 mg of ekbopride mokue CRS (0.3 pet cent);
and 5 mg of metoclopmmide hydro<hloride CRS in anhydrous - disregard limit: 0.5 times the area of the principal peak in
ethanol R and dilute to 10 mL with the same solvent. the chromatogram obtained with reference solution (a)
Ptate TLC silica gel F,,.,
pkue R.
(0.05 per cent); disregard the 2 peaks eluting within the
first 2 min.
Mobile phase concentrated ammonia R, QCtrone R, methanol R J
Chlorides
toluene R (2:14:14:70 VIVIV/V).
Maximum 100 ppm.
Apphcalwn 51JL as bands of 10 mm by 3 mm.
Prepare the solutions at the same time.
Testsolution Dissolve 0.530 g in 20.0 mL of anhydrous acetic
Drying In air. acidR, add 6 mL of dilute nitric acid R and dilute to 50.0 mL
Detection Examine in ultraviolet light at 254 nm. with water R.
System suitability Reference solution (b): Reference solutio" To 1.5 mL of o. (J(J1 M hydrochloric acid
- the chromatogram shows 2 clearly separated zones. add 20.0 mL of anhydrous acetic acid Rand 6 mL of dilute
Results The principal zone in the chromatogram obtained niln'c acid R and dilute to 50.0 mL with water R.
with the test solution is similar in position and size to the Transfer both recently prepared solutions to separate test-
principal zone in 'the chromatogram obtained with reference tubes. Add to each tube 1 mL of silver nitrate solution R2.
solution (a). Allow to stand for 5 min protected from light. Examine the
TESTS tubes laterally against a black background. Any opalescence
Solution S in the test solution is not more intense than that in the
Dissolve 1.0 g in carbon dioxide-free waterR and dilute to reference solution.
100 mL with the same solvent. Sulfates
Appearance of solution Maxiinum 100 ppm.
Solution S, examined immediately after preparation, is clear Prepare the soluUons at the same time.
(2.2.1) and colourless (2.2.2, Method 1). Testsolution Dissolve 3.00 g in 20.0 mL of glacial aua"c
pH (2.2.3) acidR, heating gently if necessary. Allow to cool and dilute
3.8 to 4.2 for solution S. to 50.0 mL with water R.
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2022 Clemastine Fumarate 1-609
Reference solution To 9 mL of slIljate standard solution

(10 ppm SO"; R1 add 6 mL of glacial acetic acidR.
Clemastine Fumarate
Into 2 test-tubes introduce 1.5 rnL of sulfale standard solution Clemastine Hydrogen Fumarate
(10 ppm SO"; R1 and add I mL of a 250 gIL solution of (Ph. Eur. monograph 1190)
barium chloride R. Shake and allow to stand for 1 min.
To one of the tubes add 15 mL of the test solution and to
the otheradd 15 mL of the reference solution. After 5 min,
any opalescence in the tube containing the test solution is not
more intense than that in the tube containing the reference
solution.
CI
)}
~
-
I
.CH3
.
0
~ .Ho,c~ co2
H CH,
H

an oven at 105°C. 460.0 14976-57-9
Sulfated ash (2.4.14) Action and use
Maximum 0.1 per cent, determined on 1.0 g. Histamine HI receptor antagonist; antihistamine.
ASSAY Preparation
Dissolve 0.400 g in 50 mL of anhydrous acetic acidR. Titrate Clemastine Tablets
with 0.1 M perchloric acid, determining the end-point
Pl>E" _
I mL of 0.1 M perchlari< acid is equivalent to 50.80 mg DEFINITION
of C 2. H 30ClN,O,. (2R) -2-[2- [( I R)-I-(4-Chlorophenyl)-I-phenylethoxy]ethyl]-I-
methylpyrrolidine hydrogen (E)-bUlenedioate.
STORAGE
Protected from ligbr. Content
IMPURITIES
Other deteaable impurities (thefollowing substances would, if CHARACTERS
present at a sufficient Ieoel, be detected by oneor other of the tests Appearance
in the monograph. They are limited by me general acceptance White or almost white, crystalline powder.
cruetion for other/unspecified impurities andlor by thegeneral Solubility
monog7aph Substances for pharmaceutical use (2034). II is Very slightly soluble in water, sparingly soluble in ethanol
therefore not necessary to identify these impwities for (70 per cent VIJI), practically insoluble in heptane.
demonstration of compliance. See also 5.10. Control of impumies
IDENTIFICATION
in substances for pharmaceutical use) A, B, C.
Comparison demastme fumarate CRS.
A. 4-amino-5-chJoro-2-methoxybenzoic acid, C. Thin-layer chromatography (2.2.27).
solvent.
Reference solution Dissolve 20.0 mg of demastine
B. I-benzylpiperidin-4-amine, fumanue CRS in methanol R and dilute to 10.0 mL with the
same solvent.
Mobile phase concentrated ammonia R) mahand R,
te<rahydrofuran R (1:20:80 VIVIJI).
Applicaliim 5 ~L.
Deudopmem Over 2/3 of the plate.
C. 4-amino-N-( I-benzylpiperidin-4-yl)-2-methoxybemamide. Drying In a current of cold airfor 5 min.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ pu"
Deuaion Spray with a freshly prepared mixture of 1 volume
of potassium iodobismuthate solmion Rand 10 volumes of dilute
acetic acid R and then with dilute hydrogen peroxide solution R;
cover the plate immediately with a glassplate of the same
size and examine the chromatograms after 2 min.
the principal Spotin the chromatogram obtained with the
reference solution.
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1-610 Clemastine Fumarate 2022
Test solution Dissolve 40 mg of the substance to be Tim, Mobile phase A Mobile phase B
(min) (per cent VIP) (per cent I'll')
solvent. 0-3 '5 55
3 - 23 45 ..... 5 55 ..... 95
Reference solution Dissolve 50 mg ofjumarie acid CRS in
23·26 5 95
ethanol (96 per cenl) R and dilute to 10.0 mL with the same
solvent.
Plate TLC silica gelplate R. FWw rate 0.8 mllmin.
Mobile phase walerR, anhydrous lonnie acidR, di-isopropyl Detection Spectrophotometer at 225 nm,
etherR (5:25:70 VIV/lI). Injection 90 ~L.
Applicalwn 5 ~L. Identification of impun'ties Use the chromatogram supplied
Development Over 213 of the plate. with clemastine for system suitability CRS and the
Drying At 100-105 °C for 30 min and allow to cool. identify the peak due to impurity B.
•." Detection Spray with a 16 gIL solution of PDULSsium Relative retention With reference to clemastine (retention
permanganate R and examine in daylight.
=
time about 17 min): fumaric acid = about 0.1;
Results The principal spot with the highest R p value in the impurity B = about 0.9.
chromatogram obtained with the test solution is similar in
position, colour and size to the principal spot in the
- resolution: minimum 2.0 between the peaks due £0
chromatogram obtained with the reference solution.
impurity Band c1emastine.
TESTS Calculation of percentage contents:
Solution S - for each impurity, use the concentration of clemastine
Dissolve 0.500 g in methanolR and dilute to 50.0 mL with fumarate in reference solution (a).
the same solvent. Limits:
Appearance of solution - unspecified impurities: for each impurity, maximum
Solution S is clear (2.2.1) and not more intensely coloured 0.10 per cent;
than reference solution BY7 (2.2.2, Method 11). - total: maximum 0.2 per cent;
pH (2.2.3) - reporting threshold: 0.05 per cent; disregard the peak due to
3.2 to 4.2. fumaric acid.
Suspend 1.0 g in 10 mL of carbon dioxide-free wow R. Loss on drying (2.2.32)
Specific optical rotation '(2.2. 7) Maximum 0.5 per cent, determined on 1.000 g by drying in
+ 15.0 to + 18.0 (dried substance), determined on
solution S. Snlfated ash (2.4.14)
Related substances
Liquid chromatography (2.2.29). Prepare the so/ww", ASSAY
immedial<!y before use. Dissolve 0.350 g in 60 mL of anhydrous acetic acid R. Titrate
Phosphate buffersduuon pH 7.1 Mix 1.9 volumes of a with 0.1 M perch/on'c acid, determining the end-point
138 gIL solution of sodium dilrydrogen phosphate potentiometrically (2.2.2U).
monohydrate R, 6.8 volumes of an 89 gIL solution of disodium I mL of 0.1 M perchlori< acid is equivalent to 46.00 mg
hydrogen phosphate dihydrate Rand 91.3 volumes of waterfor of C"H30CINO,.
chromarography R.
IMPURlTIES
Solvent mixture acetonitrile R1, waterfor chromatography R Other detectable impurities (the fo/lowing substances would, if
(20:80 VW). present at a sufficient level, be detected by one or otherof the teus
Test solution Dissolve 10 mg of the substance to be in the monograph. They are limited by the general acceptance
examined in 30 mL of the solvent mixture with the aid of criterion for other/unspecified impurities and/or by the general
ultrasound and dilute to 50.0 mL with the solvent mixture. monograph Substances for phormaceuucal use (2034). II is
Reference so/ution (a) Dilute 1.0 mL of the test solution to therefore not neteSsory to identify these impurities for
100.0 mL with the solvent mixture. Dilute 1.0 mL of this demonstration ofcompliance. See also5.10. Control of impwities
solution to 10.0 mL with the solvent mixture. in substances for pharmaceutical use) A, B, C.
f}
clemastine for system suilability CRS (containing impurity B) in ~
1.0 mL of the solvent mixture with the aid of ultrasound for
about 5 min. ,y-
-
:
CH
O~N;
3 ~-o andepimeralN
Co/umn: I it CH,
- size: 1 = 0.15 m, 0 = 4.6 mm; CI "'"
- suuionasy phose: end-eapped ethylene-bridged polar-embedded
octadecy/silyl silica gelfor chromarography (hybrid material) R A. (IRS,2R)-2-[2-[(lR}-I-( 4-cWorophenyl)-I-
(3.5 um); phenylethoxy] ethyI]-I-methylpyrrotidine l-oxide,
Mabile phose:
- mabile phoseA: phosphate buffer solution pH 7.1;
- mobile phase B: phosphate buffer solution pH 7.1,
acetonitrile Rl (40:60 VIII);
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2022 Clenbuterol Hydrochloride 1-611
Reference solution Dissolve 10 mg of clenbuterol

hydrochloride CRS in 10 mL of methanol R.
Plate TLC silica gel FZ54 pkue R.
Mobile phase ammonia R, anhydrous ethanol R, toluene R
(0.15:10:15 VIVIV).
Application I0 ~L.
B. 4-[ (IR)-I-(4-chlorophenyl)-I-phenylethoxy]-I-
methylazepane, Development Over a path of 10 em.
Drying In air.
Detection Spray with a 10 gIL solution of sodium min'te R in
C,
;}
"'" I
"-
- '. ,
.
CH
OH
andenanliorner
1 M hydrochlori< acid and dip after 10 min in a 4 gIL solution
of naphthylethY/enediamine dihydrochloride R in methanol R.
Allow to dry in air.
with the test solution is similar in position, colourand size to
C. (IRS)-I-( 4-chlorophenyl)-I-phenylethanol. the principal spot in the chromatogram obtained with the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ !'hE" reference solution.
TESTS
Soludon S
Dissolve 0.5 g in 10 mL of carbon dioxide-free water R.
Clenbuterol Hydrochloride Appearance of solution
(ph. Bur. monograph 1409) Solution S is not more opalescent than reference
c'0"
suspension IT (2.2.1) and not more intenselycoloured than
H OH reference solution Y6 (2.2.2, Method II).
"- I
~)(CH,
H,C CH,
and enenuoner • Hel
pH (2.2.3)
H,N
Optical rntatlon (2.2.7)
C,
-0.10° to + 0.10°.
313.7 21898-19-1
same solvent. Filter if necessary.
Action and use Related substances
Betaj-adrenoceptor agonist; bronchodilator. Liquid chromatography (2.2.29).
Preparations Test solution Disperse 100.0 mg of the substance to be
Clenhuterol Granules (VEl) examined in the mobile phase and dilute to 50.0 mL with
Clenbuterol Injection (VEl) the mobile phase.
Clenbuterol Oral Solution (VEl) Reference solution (a) Dilute 0.1 mL of the test solutionto
100.0 mL with warer R.
!'hE" _
Reference solutWn (b) Dissolve 5 mg of clenbuterol
DEFINITION impurity B CRS in 10 mL of the mohile phase, add 2.5 mL
( I RS)-I-(4-Amino- 3,5-dichlorophenyl) -2- [(I, I-dimethylethyl) of the test solution and dilute to 25.0 mL with the mobile
amino]ethanol hydrochloride. phase.
Golumn:
Content
- size: 1= 0.125 m, 0 = 4 mm,
- stationary phase: end-capped octadecy/siryl silica gelfor
CHARACTERS chromatography R (5 urn),
Appearance - temperature: 40°C.
White or almost white, crystalline powder. Mobile phase Mix 200 volumes of acetonitrile R, 200 volumes
Solubility of methanol Rand 600 volumes of a solution prepared as
Soluble in waterand in ethanol (96 per cent), slightly soluble follows: dissolve 3.0 g of sodium decanesulfonau Rand 5.0 g
in acetone. of potassium dihydrogen phosphate R in 900 mL of warer R,
mp adjust to pH 3.0 with dilure phosphoric acid R and dilute to
About 173 °C, with decomposition. 1000 mL with warer R.
IDENTIFICATION
First identification: A, C. Detection Spectrophotometer at 215 om.
Second identification: B, C. Injection 5 ~L.
A. Infrared absorption spectrophotometry (2.2.2,,>. Run time 1.5 times the retention time of clenbuterol.
Comparison clenbuterol hydrochloride CRS. Retention time Clenbuterol = about 29 min.
B. Thin-layer chromatography (2.2.27). System suitability Reference solution (b):
Test solution Dissolve 10 mg of the substance [0 be
impurity Band clenbuterol.
examined in 10 mL of methanol R.
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1-612 Clindamycin Hydrochloride 2022
Limits:
obtained with referencesolution (a) (0.1 per cent),
- any other impun",y: for each impurity, not more than the
with reference solution (a) (0.1 per cent), F. (IRS)-I-(4-amino-3-bromo-5-cWorophenyl)-2-[(I,I-
- taal: not more than twice the area of the principal peak in dimethylethyl)amino]ethanol.
the chromatogram obtained with reference solution (a) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 1'1>£<1
(0.2 per cent),
the chromatogram obtained with reference solution (3)
(0.05 per cent).
Clindamycin Hydrochloride
Water (1.5.11)
Maximum 1.0 per cent, determined on 0.500 g. (Ph. Ellr. monograph 0582)
ASSAY
Dissolve 0.250 gin 50 mL of ethanol (96 per cenV R and add • HCl
5.0 mL of 0.01 M hydro<h1ori1: acid. Titrate with 0.1 M
potentiometrically (2.2.21J). Read the volume added between
the 2 points of inflexion.
1 mL of 0.1 M sodium hydroxide is equivalent to 31.37 mg of 461.5 21462-39-5
C 1zHI 9C1,NzO.
IMPURITIES Action and use
Lincosamide antibacterial.
Specified impuniies A, B, C, D, E, F.
Preparations
C'qCHO
. I
Clindamycin Capsules
Clindamycin Chewable Tablets
H,N '"
Clindamycin Tablets
CI
1'1>£<1 _
A. 4-amino-3,5-dichlorobenzaldehyde,
DEFINTI10N
H,N
c'¢1
,
o
~XCH3
H3C CH3
Methyl 7-chloro-6,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpyrrolidin-2-yl)carbonyl]amino]-I-thio-L-thre<>-«-D-
galau.o-octopyranoside hydrochloride. It contains a variable
quantity of water.
Cl Semi-synthetic product derived from a fermentation product.
Content
B. 1-(4-amino-3,5-dicWorophenyl)-2-[(I, 1- 92.0 per cent to 102.0 per cent (anhydrous substance).
dimethylethyl)amino]ethanone,
CHARACTERS
Appearance
White or almost white, crystalline powder, slightly
hygroscopic.
Solubility .
Very soluble in water, slightly soluble in ethanol
(96 per cent).
C. 1-(4-amino-3,5-dicWorophenyl)ethanone,
IDENTIFICATION
First identification: A, D.
Second identijicatwn: B, C, D.
Comparison clindamycin hydrochlotide CRS.
D. 1-(4-aminophenyl)ethanone,
Test solution Dissolve 10 mg of the substance to he
solvent.
Reference solutio" (a) Dissolve 10 mg of dindamycin
same solvent.
E. 1-(4-amino-3,5-dicWorophenyl)-2-bromoethanone,
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2022 Clindamycin Hydrochloride 1-613
Reference solution (b) Dissolve 10 mg of dindamycin Systemsuitability Reference solution (a):

hydrochloride GRS and 10 mg of lincomycin hydrochloride GRS - resolution: minimum 3.0 between the peaks due to
in methanol R and dilute to 10 mL with the same solvent. impurities Band C; minimum 2.0 between the peaks due
Plate TLG silica gel G plate R. to impuriry C and c1indamycin.
Mobile phase Mix 19 volwnes of 2-propanol R, 38 volwnes Calculation of percentage contents:
of a 150 gIL solution of ammonium autau R adjusted to - for each impurity, use the concentration of clindamycin
pH 9.6 with ammonia R, and 43 volumes of emyl acetate R. hydrochloride in reference solution (b).
Use the upper layer. Limits:
Application 5 ~L. - impurity C: maximum 4.0 per cent;
- impun·'Y B: maximum 2.0 per cent,
- attYother impurity: for each impurity) maximum
Drying In air. 0.5 per cent;
Detection Spray with a I gIL.solution of potassium - total: maximwn 6.0 per cent;
pennanganate R. - reponing threshold: 0.05 per cent.
System suitability The chromatogram obtained with Water (2.5.12)
reference solution (b) shows 2 clearly separated spots. 3.0 per cent (Q 6.0 per cent) determined on 0.500 g.
Results The principal spot in the chromatogram obtained Sulfated ash (2.4. H)
with the test solution is similar in position, colour and size to Maximum 0.5 per cent, determined on 1.0 g.
. the principal spot in the chromatogram obtained with
reference solution (a). ASSAY
C. Dissolve about 10 mg in 2 mL of dilute hydrochloric acidR Liquid cbromatography (2.2.29) as described in the test for
and heat on a water-bath for 3 min. Add 3 mL of sodium related substances with the following modification.
carbonate solution Rand 1 mL of a 20 gIL solution of sodium Injection Test solution and reference solution (a).
nitroprusside R. A violet-red colour develops. Calculate the percentage content of CIsH34CI;zN205S taking
D. Dissolve 0.1 gin warer R and dilute to 10 mL with the into account the assigned content of dindamycin
same solvent. The solution gives reaction (a) of chlorides hydrochloride GRS.
(2.3.1). STORAGE
TESTS In an airtight container.
pH (2.2.3) IMPURITIES
3.0 to 5.0. Specified impuriries B, G.
Dissolve 1.0 g in carbon dioxide-free water R and dilute to Otherdetectable impurities (the fo/Wwing substances would, if
10 mL with the same solvent. present at a sufficient leveJ~ be detected by oneor other of the tests
Specific opdcal rotadon (2.2.7) in the monogmph. They arelimited by thegeneral acceptance
+ 135 to + 150 (anhydrous substance). criterion for olherlunspecrJied impurities. It is therefore not
Dissolve 1.000 g in water R and dilute to 25.0 mL with the neussary to identify these impurities for demonstration of
same solvent. compliance. See also 5.10. Control of impun·ties in substances for
pharmaceutical use) A, D, E, F.
Related substances
the mobile phase.
Reference solution (aJ Dissolve 50.0 mg of cJindamycin
hydrochloride GRS in the mobile phase and dilute to 50.0 mL
100.0 mL with the mobile phase. A. methyl 6,8-dideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpytroJidin-2-yl]carbonyl] amino ]-I-thio-n-erythro-«-
Go/umn:
- size: 1;;;; 0.25 01) 0 ;;:: 4.6 mm; D-galacco-octopyranoside (lincomycin),
- stationary phase: end-capped oaadeqlsilyl silica gelfor
chramawgmphy R (5 J1Ifl).
.t\.'. tH\CICH,
Mobile phase Mix 45 volumes of acewnitrile Rl and
55 volwnes of a 6.8 gIL solution of potassium dihydrogetl
phosphate R previously adjusted to pH 7.5 with a 250 giL
solution of potassium hydroxide R.
P
H····
CH,
(HO
o
~....
OH
0
OH
S,
CHJ
lnjection 20 jJL. B. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2S,4R}-4-ethyl-l-
methylpyrroJidin-2-yl]carbonyl] amino] -f-thio-r.-threo-o-o-
Rim time Twice me retention time of c1indamycin.
galact<J-octopyranoside (clindamycin B),
Relative retention With reference to c1indamycin (retention
time = about 10 min): impurity B = about 0.7i
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1-614 Clindamycin Phosphate 2022
Preparations
Benzoyl Peroxide and Clindamycin Gel
Clindamycin Gel
Clindamycin Injection
Clindamycin Lotion
Clindamycin Solution
Clindamycin Vaginal Cream
C. methyl 7-chlor<>-6,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
propyIpyrrolidin-2-yl]carbonyl] amino)-I-thio-D-erylhn>-«- Sodium FusidateTablets
D-galacto-octopyranoside (7-epiclindamycin), PIlE" _
DEFINITION
Methyl 7-chloro-S,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpyrrolidin-2-yl]carbonyl]amino)-2-0-phosphono-l-
thio-L-I1nw-«-D-galacto-octopyranoside.
Content
96.0 per cent 10 102.0 per cent (anhydrous substance),
D. methyl 6,8-dideoxy-6-[[[(2S.4R)-I-methyl-4- CHARACTERS
propylpyrrolidin-2-yl] carbonyl] amino)-I-thio-L-lhreo-«-D- Appearance
galacco-octopyranoside (7-epilincomycin), WhIte or almost white, slightly hygroscopic powder.
Solubll1ly
Freelysoluble in water, vecyslightly soluble in ethanol
IDENTIFICATION
Firstidentification: A, D.
Second identificarion: B, C, D.
E. methyl (5R)-5-[ (IS,2S)-2-chloro-I-[[(4Z)-I-methyl-4- A. Infrared absorption spectrophotometry (2.2.24).
propylidene-L-prolyl)amino)propyl)-I-thio-fl-L- Comparison dindamycin phosphate CRS.
arabinopyranoside, If the spectra obtained in the solid state show differences,
dissolve 50 mg of the substance to be examined and the
reference substance separately in 0.2 mL of waterR and heat
until completelydissolved. Evaporate to dryness under
reduced pressure, dry the residues at 100-105 °C for 2 hand
F. methyI7-chloro-6,7,8-trideoxy-6-[[[(2R,4R)-I-methyl-4- solvent.
propyIpyrrolidin-2-yl]carbonyl] amino)-1-thio-L-lhre<>-a:-0- Reference solulion (a) Dissolve 20 mg of cUndamycin
galaclo-ocwpyranoside. phosphate CRS in methatwlRand dllure to 10 mL with the
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" same solvent.
Reference solUlion (b) Dissolve 10 mg of lincomycin
hydrochloride CRS in 5 mL of reference solution (a).
Plate TLC silica gelplateR.
Clindamycin Phosphate lWobile phase glacial acetic acid R, waterR, buuznol R
(20:20:60 VIV/V).
Applicarion 5 IlL.
Deodopment Over 2/3 of the plate.
Drying At 100-105 "C for 30 min.
Detection Spray with a 1 gIL solutionof potassium
permanganate R.
- the chromatogram shows 2 principal spots.
505.0 24729-96-2
Action and use C. Dissolve ahout 10 mg in 2 mL of dilute hydrochloric acidR
Lincosamide antibacterial. and heat in a water-bath for 3 min. Add 4 mL of sodium
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2022 Clindamycin Phosphate 1-615
carbonate solution Rand 1 mL of a 20 gIL solution of sodium Relativeretention With reference to clindamycin phosphate
nitroprusside R. Prepare a standard in the same manner using (retention time = about 12 min): impurity F = abcur 0.15j
c/indamydn phosphate CRS. The colour of the test solution impurity G = about 0.19; impurity I = about 0.34;
corresponds to that of the standard. impurity B = about 0.45; impurity L =about 0.64;
D. Boil 0.1 g undera reflux condenser with a mixture of impurity J = about 1.20j impurity E = about 1.73;
5 mL of strong sodium hydroxide solution Rand 5 mL of =
water R for 90 min. Cool and add 5 mL of m"uU, acid R. System suitability Reference solution (c):
Extract with 3 quantities, each of 15 mI..., of methylene - resolution: minimum 2.0 between the peaksdue to
chloride R and discard the extracts. Filter the upper layer impurities F and Go
through a paper filter. The fih.rate gives reaction (b) of Calculation of percentage contents:
phosphates (2.3.1). - for each impurity, use the concentration of c1indamycin
TESTS phosphate in reference solution (b);
Solution S Limits:
Dissolve 1.00 g in carbon dioxide-free water R. Heat gently if - impurities B, L: for each impurity, maximum 1.0 per cent;
necessary. Cool and dilute to 25.0 mL with carbon dioxide-free - impuruies E~ F: for each impurity, maximum 0.5 per cent;
water R. - impun'ties G, I, J, K: for each impurity, maximum
Appearance of the solution 0.2 per cent;
Solution S is clear (2.2.1) and colourless (2.2.2, Me/hod 11). - unspecified impurities: for each impurity, maximum
0010 per cent;
Related substances - Ictal: maximum 2.0 per cent;
Liquid chromatography (2.2.29). _ reporting threshold: 0.05 per cent.
Water (2.5.12)
mobile phase A.
Reference solution (a) Dissolve 30.0 mg of clindamycin
Less than 0.6 IU/mgJ if intended for use in the manufacture
phosphate CRS in mobile phase A and dilute to 10.0 mL with
mobile phaseA.
20.0 mL with mobile phase A. Dilute 1.0 mL of this solution ASSAY
to 10.0 mL with mobile phase A. Liquid chromatography (2.2.29) as described in the test for
Reference solution (c:) Dissolve 3.0 mg of dindamycin related substances with the following modification.
phosphate for system suitability CRS (containing impurities B, lnjution Test solution and reference solution (a).
E, F, G, IJJJ K and L) in mobile phase A and dilute to Systemsuitability Reference solution (a):
1.0 mL with mobile phase A. - symmetry factor: maximum 3.0 for the peakdue to
Column: clindamycin phosphate.
- size: / =0.15 m, 0 = 4.6 mm; Calculate me percentage content of ClsH34CIN20sPS raking
- stationary phase: end-capped octadocylsiiyl silica gelfor into account the assigned content of dindamycin
chromatography R (5 ~m); phosphate CRS.
- temperature'; 30°C. STORAGE
Mobile phase: In an airtight container. If the substance is sterile, the
- mobile phase A: mix 21 volumesof acetonitrile Rl and container is also sterile and tamper-evident.
phosphate R previously adjusted to pH 6.0 with a 450 gIL IMPURITIES
solutionof potassium hydroxide R; Specified impurities B, E, FJ G, I, J, ~ L.
- mobile phase B: mix 40 volumes of a 13.6 gIL solution of Other detectable impurities (the following substances would, if
potassium dihydrogen phosphate R previously adjusted to present at a sufficient lwd, be detected by oneor other 0/the tests
pH 6.0 with a 450 gIL solution of potassium hydroxide R, in the monograph. They a.. limited by thegeneral acceptance
and 60 volumes of acetonitrile Rl, ctitenon for other/unspecified impwines and/or by thegeneral
monograph Substances for pharmaceutical use (2034). 1, is
Time Mobile phase A Mobile phase B there/ore not nuessary to identify these impun'ties for
(min) {per cent Vm (per cent I'm demonstration of wmplianc:e. See also 5.10. Control of impunues
0·13 100 0 in substances for pharmaceutical use) A, C~ D, H.
13 - 18 100 ---> 50 0--->50
18·39 50 50
FlOOJ rate 1.1 mUmin.

Injection 20 J.tL of the test solution and reference
with dindamycin phosphate for system suitability CRS and the A. methyl 6,8-dideoxy-6-[[[(2S,4R)-I-methyl-4-
chromatogram obtained with reference solution (c) to identify propylpyrrolidin-2-yl]carbonyl)amino]-1- thi<>-D-erythro-«-
the peaks due to impurities B, E, F, G, I, J, K and L. D-galacto-octopyranoside (lincomycin),
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1-616 Clindamycin Phosphate 2022
,yH, ~--,~H'Cl ,yH,",H ~---~HO"

H
P
H--
CH,
.H
o
(HO
HO
OH
•
0
0
CH,
S,
CH3
H"
f
H,C HO'!
(0
o
p---o
OH
0
CH,
S'CH,
'p'
HO 0
,~ o"
G. methyl 6,8-dideoxy-2,4-0-(hydroxyphosphoryl)-6-
B. methyl 7-cWoro-6,7,8-trideoxy-6-[[[(2S,4R)-4-ethyl-l- [[[(2S, 4R)-I-methyl-4-propylpyrrolidin-2-yljcarbonyl]
methylpyrrolidin-2-yl)carbonyl]amino)-2-Q--phosphono-l- amino]-I-thio-D-erymro-«-D-galaclO-ocropyranoside (2,4-
thio-L-threo-«-D-galaClO-octopyranoside (c1indamycin B phosphatidyllincomycin),
2-phosphate),
C. methyl 7-chloro-e,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4- H. methyl 7-chloro-e,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-

propylpyrrolidin-2-yljcarbonyl)aminol-3-Q--phosphono-l- propylpyrrolidin-2-yl)carbonyljamino)-2,3-di-Q--
thio-L-threo-ct-D-galacr.o-octopyranoside (clindamycin phosphono-l-thio-L-threo-ct.-D-galacto-octopyranoside
3-phosphate), (dindamycin 2,3-bisphosphate),
D. methyl 7-cWoro-6,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpyrrolidin-2-yljcarbonyl)aminoj-4-Q--phosphono-l- L methyl 7-cbloro-S,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
thio-L-threo-«-D-galaceo-octopyranoside (clindamycin propylpyrrolidin-2-yl)carbonyl)aminoj-2,4-di-Q--
4-phosphate), phosphono-l-thio-L-thre.o-a~D~galacto-octopyranoside
(dindamycin 2,4-bisphosphate),
E. methyl 7-chloro-o,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpyrrclidin-2-yl]carbonyl] aminoj-l-thiO-L-threo-«-D-
galaceo-octopyranoside (clindamycin), J. methyI7-cWoro-6,7,8-trideoxy-6-[[[(2S)-I-methyl-4-
propylidenepyrrolidin-2-yijcarbonyijaminoj-2-Q--
phosphono-l-thio-L-threo-<X-D-galacto-octopyranoside
(propylidene analog of clindamycin 2-phosphate),
F. methyI6,8-dideoxy-6-[[[(2S,4R)-I-methyl-4-
propylpyrrolidin-2-yljcarbonyljamino)-2-Q--phosphono-l-
thio-D-erythro-<t-D-galaceo-octopyranoside (lincomycin
K. 2,2'-oxybis(hydroxyphosphoryl)bis[methyl 7-cWoro-6,7,8-
2-phosphate),
trideoxy-6- [[ [(2S, 4R)-I-methyl-4-propylpyrrolidin-2-yIJ
carbonyl)amino)-I-thio-L-threo-<X-D-galaceo-octopyranoside)
(didindamycin pyrophosphate),
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2022 ClioquinoI 1-617

Preparation Discs of potassium bromide R.
Comparison clioquinol CRS.
C. Whenheated, violet fumes are produced.
D. Dissolve about I mg in 5 mL of ethanol (96 per un\! R.
Add 0.05 mL offerne chloride solution Rl. A dark green
colourdevelops.
TESTS
L. methyl 7-chloro-6,7,8-trideoxy-6-[[[(2S,4R)-I-methyl-4- Acidity or alkalinity
propylpyrrolidin-2-yl]carbonyl)amino)-2-D-phosphono-l- Shake 0.5 g with 10 mL of carbon dioxide-free water Rand
thio-D-erythro-<.J.-D-ga«Uto-octopyranoside filter. To the filtrate add 0.2 mL of phenolphthalein solution R.
(7-epiclindamycin z-phosphate). The solution is colourless. Not more than 0.5 mL of 0.01 i\tI
__________________ ~PhE«
sodium hydroxide is required to change the colourof the
indicator to pink.
Related substances
Clioquinol Test solution Dissolve 50.0 mg of the substance to be
solvent) heating gently if necessary. Dilute 10.0 mL of the
'hN~ Reference solution (a) Dissolve 20.0 mg of 5-<:hloroquinolin-B-

01 R, 10.0 mg of 5,7-dichloroquinolin-B-oI R, 5 mg of the
yv CI
substance to be examined and 10.0 mg of 5,7-diiodoquinolin-
8-01 R in methanol R, heatinggently if necessary and dilute to
C,H,CIINO 305.5 130-26-7 Reference solution (b) Dilute 1.0 mL of reference solution (a)
Action and use
Anrlbacreriaf anriprotozoal, Reference solution (c) Dilute 1.0 mL of the test solution to
Preparations
solution to 20.0 mL with me mobile phase.
Betamethasone and Clioquinol Cream
Column:
Betamethasone and Clioquinol Ointment
- size: 1= 0.15 m, 0 = 3.9 mm,
Flumetasone and Clioquinol EarDrops - stationary phase: octylsilyl silica gelfor chromatography R
Hydrocortisone and Clioquinol Cream (5 1UJl).
Hydrocortisone and Clioquinol Ointment Mobile phase Dissolve 0.50 g of sodium edeuue R in 350 mL
PhE« _ of water R, add 4.0 mL of hexylamine R and mix. Adjust to
pH 3.0 with phospho";'; acidR. Add 600 mL of methanol R
DEFINITION and dilute to 1000 mL with waterR.
s-Chloro-?-iodoquinclin-B-ol, Flow rate 1.3 mlJmin.
Content Detection Spectrophotometer at 254 om.
98.0 per cent to 102.0 per cent (dried substance). Injmiot. 20 ~L.
CHARACTERS Run time 4 times the retention time of clioquinoI.
Appearance Relative retention With reference to clioquinol (retention
Almost white, light yellow, brownish-yellow or yellowish-grey time = about 10 min): impurity A =about 0.4;
powder. impurity B = about 0.7; impurity C = about 1.3.
Solubility System suitability Reference solution (a):
Practically insoluble in water, sparingly soluble in methylene - resolution: minimum 3.0 between the peaks due to
chloride, very slightly soluble or slightly soluble in ethanol clioquinol and impurity C.
(96 per cent). Limits:
IDENTIFICATION - impurity A: not more than the area of the corresponding
Firsl idendfication: B. peak in the chromatogram obtained with reference
solution (b) (2.0 per cent),
Second identification: A, CJ D.
- impun'ty B: not more than the area of the corresponding
A. Dissolve 40.0 mg in methanol R and dilute to 100.0 mL
with the same solvent. Dilute 10.0 mL to 100.0 mL with solution (b) (1.0 per cent),
methanol R (solution A). Examined between 280 nm and - impun'ty C: not more than the area of the corresponding
350 run (2.2.25), solution A shows an absorption maximum peak in the chromatogram obtained with reference
at 321 nm. Dilute 10.0 mL of solution A to 100.0 mL with solution (b) (1.0 per cent),
methanol R (solution B). Examined between 230 nm and - unspecified impun'ties: for each impurity, not more than
280 run, solutionB shows an absorption maximum at twice the area of the principal peak in the chromatogram
255 run. The specific absorbance at this absorption obtained with reference solution (c) (0.10 per cent),
maximum is 1530 to 1660.
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1-618 Clobazam 2022
- total of me nominal contents of ;mpun"ties A, B, C and

Clobazam ***
unspecified impumies: maximum 3.0 per cent,
disregard limit: the area of the principal peak in me
*** ***
- (Ph. Eur. monograph 1974) ***
(0.05 per cent).
Halides
Maximum 140 ppm, expressed as chlorides.
Shake 0.5 g with 25 mL of water R for I min and filter.
To the filtrate add 0.5 mL of dilute nitn' acid Rand 0.5 mL
of silver nitrate solution RZ. Allow to standfor 5 min.
Any opalescence is not more intense than that in a standard
prepared at the same time by adding 0.5 mL of silvernitrate
solution R2 to 25 mL of water R containing 0.2 mL of O. 01 M
300.7 22316-47-8
hydrochloric acid and 0.5 mL of dilute nitric acid R.
Loss on drying (2.2.32) Action and use
Maximum 0.5 per cent, determined on 1.000 g by drying in Benzodiazepine.
a desiccator at a pressure not exceeding 0.7 kPa for 24 h. Preparations
Sulfated ash (2.4.14) Clobazam Oral Suspension
Maximum 0.1 per cent, determined on 1.0 g. Clobazam Tablets
ASSAY Pflf(r _
Dissolve 0.200 g in 20 mL of acetic anhydride R and add
30 mL of glacial acetic acid R. Titrate with 0.1 M perch/oric DEFINITION
add, determining the end-point potentiometrically (2.2.20). 7-Chioro-I-methyl-5-phenyl-I,5-dihydro-3H-I ,5-
I mL of 0.1 M pcrchlaric acid is equivalent to 30.55 mg of benzodiazepine-2,4-dione.
total quinolines, calculated as clioquinot. Content
Specified impurities A, B; G.
Solubility
Slighdy soluble in water, freely soluble in methylene chloride,
sparingly soluble in alcohol.
IDENTIFICATION
Gomparison Ph. Eur. reference spectrum of ciobazam.
A. 5-cWoroquinolin-8-o1, TESTS
Related substances
Test solution Dissolve 10.0 mg of me substance to be
examined in the mobile phase and dilute to 50.0 rnL with
the mobilephase.
Reference solution (a) Dissolve 5.0 mg of clabazam
impurily A CRS in the mobile phase and dilute to 50.0 mL
B. 5,7-dichloroquinolin-8-o1, with the mobile phase. Dilute 1.0 mL of the solution to
OH Reference solution (b) Dissolve 5 mg of chlordiazepoxide CRS
''¢O I
and 5 mg of donczepam GRS in the mobile phase and dilute
to 50 mL with the mobile phase. Dilute I mL of the solution
to 100 mL with the mobile phase.
Reference sdsuion (c) Dilute 1.0 mL of the test solution to
C.5,7-dtiodoquinolin-8-01. Golumn:
_____ ~ ~ PhE" - size: 1= 0.25 m, 0 = 4.6 mm,
- stationary phase: octad«y/si/yl silica gelfor chromatography R
(5 um),
Mobile phase acetonim7e R, water R (40:60 VIV).
Flaw rau I rnUmin.
lnjection 20 ~L.
Run time 5 times the retention time of clobazam,
Retention rime Clobazam:::: about 15 min.
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2022 Clobetasol Propionate 1-619
System suitabilisy Reference solution (b):

- resolution: minimum 1.3 between the peaks due (0
chlordiazepoxide and clonazepam.
Limits:
- impun"ty A: not more than me area of the principal peak
(0.5 per cent),
- any other impun"ty; not more than 0.4 times the area of the
principal peak in the chromatogram obtained with D. 7-chloro-l,3,3-trimethyl-5-phenyl-1,5-dihydro-3H-I ,5-
reference solution (e) (0.2 per cent), benzodiazepine-2,4-d.ione,
- total of other impurities: not more than twice the area of the
reference solution (e) (1.0 per cent),
(0.05 per cent).
an oven at 105 DC.
Sulfated ash (2.4.14) E. N-[4-ehloro-2-(phenylamino)phenyl]-N-methylace,amide,
Maximum 0.1 per cent, determined on me residue obtained
in the test for loss on drying.
ASSAY
Dissolve 50.0 mg in alcohol R and dilute to 100.0 mL with
the same solvent. Dilute 2.0 mL of the solution to 250.0 mL
with akoho! R. Measure the absorbance (2.2.25) at the
maximwn at 232 nm.
Calculate the content of Cl~13CIN202 taking the specific
absorbance [Q he 1380. F. methyl 3-[[4-chloro-2-(phenylamino)phenyl]
IMPURITIES methylarnino]-3-oxopropanoate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PbE"
Clobetasol Propionate ****

** *
(ph. Eur. monograph 2127) *****
A. 7-chloro-5-phenyl-I,5-dihydro-3H-I,5-benzodiazepine-
2,4-dione,
C"H32CIFO, 467.0 25122-46-7
Action and use

Glucocorticoid.
B. l-methyl-5-phenyl-I,5-dihydro-3H-I,5-benzodiazepine- Preparations
2,4-dione, Clobetasol Cream
Clobetaaol Cutaneous Foam
Cloberasol Ointment
Clobetasol Scalp Application
Clobetasol Shampoo
andenanliomer
PbE" --'---_ _
DEFINITION
21-Chloro-9-fluoro-ll ~-hydroxy-16Il-methyl-3,20
dioxopregna-l,4-dien-17-yl propanoate,
C. (3RS)-7-ehloro-l,3-dimethyl-5-phenyl-I,5-dihydro-3H-
l,5-benzodiazepine-2,4-dione, Content
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1-620 ClobetasoI Propionate 2022
CHARACTERS System suitability Reference solution (b):

Appearance - resolution: minimum 2.0 between the peaks due to
White or almost white, crystalline powder. clobetasol propionate and impurity J.
Solubility Calculation ofpercemage rontents:
Practically insoluble in water, freely soluble in acetone, - correction factor. multiply the peak area of impurity B by
sparingly soluble in ethanol (96 per cent). 0.6;
- for each impurity, use the concentration of c1obetasol
IDENTIFICATION propionate in reference solution (d).
Limits:
Comparison clobetasol propionate CRS. - impuniies BJ E: foreach impurity, maximum 0.3 per cent;
TESTS - impwilies A, D: for each impurity, maximum 0.2 per cent;
Specific optical rotation (2.2.7) - unspecified impurities: for each impurity) maximum
+ 112 to + 118 (dried substance). 0.10 per cent;
Dissolve 0.250 g in ace«me R and dilute to 25.0 mL with the - tola/: maximum 1.0 per cent;
same solvent. - reporting threshold: 0.05 per cent.
Liquid chromatography (2.2.29). Maximum 0.5 per cent, determined on 1.000 g by drying in
Test solution (aJ Dissolve 20.0 mg of the substance to be
examined in the mobile phase and dilute to 20.0 mL with Sulfated ash (2.4.14)
the mobile phase. Maximum 0.1 per cent) determined on 1.0 g.
Test schuion (b) Dissolve 20.0 mg of the substance to he ASSAY
examined in the mobilephase and dilute to JOO.O mL with Liquid chromatography (2.2.29) as described in the test for
the mobile phase. related substances with the following modification.
Reference solution (a) Dissolve 20.0 mg of dobetasol InJection Test solution (b) and reference solution (a).
propionate CRS in the mobile phase and dilute to 100.0 mL Calculate the percentage content of C2sH32ClFOs taking into
with the mobile phase. account the assigned content of clobetasol propitmau CRS.
STORAGE
clobetasol impurityJ CRS in 2 mL of the mobile phase.
To 0.5 mL of the solution add 0.5 mL of test solution (b)
and dilute to 20 mL with the mobile phase. IMPURITIES
Reference solution (c) Dissolve the contents of a vial of Specified impurisies A, B, D, E.
dobetasol propionate for peak identification CRS (containing Ocher detectable impun'ries (the following substances would, if
impurities A, B, D and E) in 2 mL of the mobile phase. present at a sufficient level, be deucud by oneor other of the tests
Reference solution (d) Dilute 1.0 mL of test solution (a) to in themonograph. They are limited by the gtneralacceptance
100.0 mL with the mobile phase. Dilute 1.0 mL of this criterion for ocherlunspecified impurities and/or by thegeneral
solutionto 10.0 mL with the mobilephase. monograph Substances for pharmaceuti<a1 use (2034). It is
Column:
demonstration of camp/iance. See also 5.10. Control of impurities
- size: 1= 0.15 m, 0 = 4.6 mm;
in substances for pharmaceutical use) CJ FJ G, H) I, J, K.
- stalionary phase: end-capped ocradecy/silyl silica gelfor
chromatography R (5 J1III); OH
- temperature: 30 "C. o
Mobile pkase Mix 10 volumes of methanol R 1, 42.5 volumes ...',/'oo.,_ O~CH3
of a 7.85 gIL solution of sodium dihydrogen phosphate
\ CH3
monohydrate R adjusted to pH 5.5 with a 100 gIL solution of ,p""- II -"'" 'A"-./ H
sodium hydroxide R and 47.5 volumes of acetonitrile for
chromatography R. o
Detection Spectrophotometer at 240 nm. A. 9-lIuoro-11p,2I-<1ihydroxy-16p-methyl-3,20-dioxopregna-
IA-dien-17-yl propanoate (betamethasone 17-propionate»)
Injection 10 J,tL of test solution (a) and reference
Run lime 3 times the retention time of c1obetasol
propionate.
Identification of impun·ties Use the chromatogram obtained
withreference solution (b) to identify the peakdue to
impurity J; use the chromatogram supplied with cIobetasol
propionare for peak identification CRS and the chromatogram o
obtained with reference solution (c) to identify the peaks due
to impurities A) B) D and E. B. 21-chloro-9-lIuoro-ll p-hydcoxy-16-methylpregna-l,4, 16-
Rdative retention With reference to c1obetasol propionate triene-3)20-dione,
(retention time = about 11 min): impurity A = about 0.4;
impurity B = about 0.6; impurity J = about 1.1;
= =
impurity D about 1.2; impurity E about 2.1.
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2022 Clobetasone Butyrate 1-621
o
C. 21-chloro-9-lIuoro-11 p-hydroxy-I6a-methyl-3,2o-
dioxopregna-I,4-dien-17-yl propanoate, I. 9-lIuoro-11 p-hydroxy-16p-methyl-21-
[(methanesulfonyl)oxyJ-3,20-dioxopregna-1 ,4-dien-17-yl
pmpanoate,
CI
D. 21-chloro-9-lIuoro-ll p-hydroxy-16p-methyl-3,20-
dioxopregn-4-en-17-yl propanoate (l,2-dihydroclobetasol o
17-propionate),
J. (17 R)-4'-chloro-5'-ethyl-9-lIuoro-ll/}-hydroxy-16P-
o CI methylsplro]androsta-I ,4-diene-17,2'-furan]-3,3'-dione
o (17a-spiro compound),
CH 3 o~CH3
', CH 3
o
H
O~CH3
o
E. 21-chloro-16/}-methyl-3,20-dioxopregna-I,4-dien-17-yl
propancate,
o
K. 9-lIuoro-11 p, 17-dihydroxy-16p-methyl-3,20-dioxopregna-
l,4-dien-21-yl propanoate (betamethasone 21-propionate).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
F. 9_lIuoro_llp_hydroxy_16/}-methyl_3_oxopregna_I,4,17
***
(20)-trien-21-oic acid, Clobetasone Butyrate •• *•
CI
*****
G. 21-chloro-9-lIuoro-ll p, 17-dihydroxy-16p-methylpregna- o
1,4-d.iene-3,20-dione (clobetasol),
C,J!3,CIFO, 479.0 25122-57-0
Action and use

Glucocorticoid.
Preparations
Clobetasone Cream
o Cloberasone Ointment
PhE" _
H. 9-lIuoro-11 p-hydroxy-16p-methyl-3,20-dioxopregna-I,4-
dien-I? -yl propanoate, DEFINITION
21-Chloro-9-lIuoro-16p-methyl-3, II ,20-trioxopregna-I,4-
dien-17-yJ butanoate.
Content
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1-622 Clobetasone Butyrate 2022
CHARACTERS Limits:
Appearance - unspecified impurities: for each impurity, not more than the
White or almost white powder. area of the principal peak in the chromatogram obtained
Solubillty with reference solution (b) (0.10 per cent);
Practically insoluble in water, freely soluble in acetone and in - total: not more than 5 times the area of the principal peak
methylene chloride, slightly soluble in ethanol (96 per cent), in the chromatogram obtained with reference solution (b)
(0.5 per cent);
mp - disregard limit: 0.5 times the area of the principal peak in
About 178 'C. the chromatogram obtained with reference solution (b)
IDENTIFICATION (0.05 per cent).
Infrared absorption spectrophotometry (2.2.24). Loss on drying (2.2.32)
Comparison clobeUlSone butyrate CRS. Maximum 0.5 per cent, determined on 1.000 g by drying in
TESTS an oven at 105°C.
Specific optical rotation (2.2.7) ASSAY
+ 131 to + 138 (dried substance). Dissolve 20.0 mg in ethanol (96 per cen'" R and dilute to
Dissolve 0.250 g in ethanol R1 and dilute to 25.0 mL with 100.0 mL with the same solvent. Dilute 5.0 mL of the
the same solvent. solution to 50.0 mL with ethanol (96 per <en'" R. Measure the
absorbance (2.2.25) at the absorption maximum at 235 om.
Related substances
Liquid chromatography (2.2.29). Prepare the solutions Calcuiate the content of C,.H,.CIFO" taking the specific
immediately before use. absorbance to be 327.
Solvent mixture anhydrous formic acid R, aaumilrile R, STORAGE
waterR (0.1:43:57 VIVIV). Protected from light.
Test solution Dissolve 65 mg of the substance to be IMPURITIES
examined in 5.0 mL of aceu>nitrile R and dilute to 25.0 mL Other detectable impurities (thefollowing substances would, if
with the solvent mixture. present at a sufficient level.. be detected by one or other 0/ the tests
Reference solution (a) Dissolve 13 mg of dobetasone butyrate in the monograph. They areli'mired by the general acceptance
for system suitability CRS (containing impurity F) in 1 mL of criterion for otherlunspecified impurities and/or ry the general
acetonitrile R and dilute to 5.0 mL with the solvent mixture, monograph Substances for pharmaceutical use (2034). It is
Reference solution (b) Dilute 1.0 mL of the test solution to therefore notnecessary to identify these ;mpuniies for
100.0 mL with the solveni mixture. Dilute 1.0 mL of this demonstration of compliance. See auo 5.10. Conrro/ of impuniies
solution to 10.0 mL with the solvent mixture. in substances for phannauutiall use) A J CJ D J E.. F.. G.. H.. 1.
Column:
- size: 1= 0.15 m, 0 = 4.6 mm;
- stationary phase: end-capped oetade<y/silyl silica gelfor
chromategraphy R (3.5 um),
Mobile phase:
- mobile phase A: anhydrous form;'; acid R, water R o
(0.1:99.9 VII');
-r- mobile phase B: anhydrous form;'; acid R, acetenitrik R A. 21-chloro-9-f1uoro-17-hydroxy-Ifijl-methylpregna-Ld-
(0.1:99.9 VII'); . diene-3,1l,20-trione (clobetasone),
'ome Mobile phase A MobUe phase B

57 43
57 -0 43 43 -0 57

o
Detection Spectrophotometer at 241 run. H
Injection 10 ~L.
C. 21-chloro-9-f1uoro-16jl-methyl-3,ll,20-tnoxopregn-l-en-
Identificotion of impurities Use the chromatogram supplied
l7-yl butanoate (4,5-dihydroclobetasone butyrate),
with dobetasone butyrate for system suitability CRS and the
chromatogram obtainedwith reference solution (a) to CI
identify the peak due to impurity F. o
Relative retention With reference to c1obetasone butyrate
(retention time = about 14 min): impurity F = about 0.9.
"'O~CH3
\ CH3
System suitability: H
impurity F and c1obetasone butyrate in the chromatogram o
obtained with reference solution(a);
- signal-to-noise ratio: minimum 10 for the principal peakin D. 2-bromo-21-cWoro-9-f1uoro-16jl-methyl-3,11,20-
the chromatogram obtained with reference solution (b). trioxopregna-Lq-dien- 17-yl butanoate
(2-bromoclobetasone butyrate),
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2022 Clofazimine 1-623
Clofazimine ***
*** ***
E. 2 l-chloro-9-f1uoro-16Il-methyl-3, 11,20-trioxopregn-4-en-
17-yl butanoate (l,2-dihydroclobetasone butyrate),
473.4 2030-63-9
o Action and use
Antileprosy drug.
F. zf-chloro-s-Suoro-tec-methvl- 3,11,20-trioxopregna-I,4- Preparation
dien-17-yl butanoate (l6a,:'methyl c1obetasone butyrate),
Clofazimine Capsules
PhE<r _
DEFINITION
N,5-Bis(4-chlorophenyl)-3-[(I-methylethyl)imino]-3,5-
dihydrophenazin-2-amine.
Content
99.0 per cent £0 101.0 per cent (dried substance).
o CHARACTERS
Appearance
G. 9-fiuoro-16~-methyl-3,11,20-trioxo-21 Reddish-brown, fine powder.
(propanoyloxy)pregna-I,4-dien-17-yl butanoate, SolublIlty
Practically insoluble in water, soluble in methylene chloride,
CI
o very slightly soluble in ethanol (96 per cent).
O~CH3
....
, CH3 IDENTIFICATION
",-,....-"-'I/'--'- 'H First identification: A.
Second idendficosion: B, C.
o
H. 21-chloro-9-fiuoro-I6~-methyl-3,11,20-trioxopregna-I,4 Comparison cWfazimine CRS.
dien-17-yl propanoate (17-0-propionyl c1obetasone), If the spectra obtained in the solid stateshow differences,
substance separately in methylene chloride R, evaporate to
examined in -methylene chloride R and dilute to 10 mL with
o the same solvent.
Reference solution Dissolve 10 mg of dofazimine CRS in
I. zt-chloro-s-ffuoro-Iep-methyl-3,11,20-trioxopregna-I,4- methylene chloride R and dilute to 10 mL with the same
dien-17-yI2-methylpropanoate (17-0-isobutyryl solvent.
clobetasone). Pial< TLC silica gel GFm pkue R.
- ~ PhE<r Mobile phase propanol R, methylene chloride R (6:85 VIV).
Applicatwn 5 JJL.
First development Over 2/3 of the plate.
Drying Horizontally in airfor 5 min.
Second deoelopment Over 213 of the plate.
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1-624 Clofibrate 2022
principal spot in the chromatogram obtained with the ASSAY

reference solution. Dissolve 0.400 gin 5 mL of methyl. ..e chloride R and add
C. Dissolve 2 mg in 3 mL of acetone R and add 0.1 mL of 20 mL of acetone R and 5 mL of anhydrous acetic acid R.
hydrochloric acid R. An intenseviolet colour is produced. Titratewith 0.1 M perchioric acid, determining the end-point
Add 0.5 mL of a 200 gIL solution of sodium hydroxide R; the potentiometrically (2.2.20).
colour changes to orange-red. I mL of 0.1 M perchlotic acid is equivalent to 47.34 mg
TESTS of C27H22C12N4"
Liquid chromatography (2.2.29). Prepare the solution' Specified impun'ties A, B.
TtSt solution Dissolve 50 mg of the substance to be
examined in the mobile phase and dilute to 100 mL with the
mobile phase.
Reference solution (b) Dissolve 5.0 mg of cIofazimine for
system suitability CRS in the mobile phase and dilute to A. N,5-bis(4-chlorophenyl)-3-imino-3,5-dihydrophenazin-2-
10.0 mL with the mobile phase. amine,
Golumn:
- size: / = 0.25 m, 0 = 4.6 mm,
- stationary phase: lXtylsi/y1 silica gelfor chromawgraphy R
(5 urn}.
Mobik phase Dissolve 2.25 g of sodium laurilsulfate R, 0.85 g
of tetrabutylammomum hydrogen sulfate Rand 0.885 g of
disodium hydrogen phosphate dolkcaitydrate R in waterR.
Adjust to pH 3.0 with dilute phosphoric acid R and dilute to
500 mL with water R. Mix 35 volumes of this solution and
65 volumes of acetonitrile R.
Flow rate 1 mUmin. B. 5-(4-chlorophenyl)-3-[(I-methylethyl)iminol-N-phenyl-
Detection Spectrophotometer at 280 nm. 3,5-d.ihydrophenazin-2-amine.
Injection 20~. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ IMm
Run time 3 times the retention time of clofazimine.
with dofazjmjnefor system suitability GRS to identify the peak
due to impurity B. Clofibrate
Relauve retention With reference to clofazimine (retention
time > about 15 min): impurity A;;;: about 0.7j (ph. Eur. monograph 01/8)
=
- resolution: baseline separation between the peaksdue to
impurity B and c1ofazimine.
Limits:
- impun"ty A: not more than the area of the principal peak 242.7 617-07-0
(0.1 pet cent), Acdon and use
- impurity B: not more than 3 times the area of the Fibrate; lipid-regulating drug.
principal peak in the chromatogram obtained with PhEm _
- any other impuniy: for each impurity, not more than the DEFINITION
area of the principal peak in the chromatogram obtained EthyI2-(4-chlorophenoxy)-2-metbylpropionate.
with reference solution (a) (0.1 per cent), CHARACTERS
Appearance
Clear, almostcolourless liquid.
(0.5 per cent),
- disregard limit: 0.5 times the area of the principal peak in Solubility
the chromatogram obtainedwith reference solution (a) Very slightly soluble in water, miscible with ethanol
(0.05 per cent). (96 per cent).
Loss on drying (2.2.12) IDENTIFICATION
Maximum 0.5 per cent, determined on 1.000 g by drying in A. Infrared absorption spectrophotometry (2.2.24).
an oven at 105 DC. Comparison clofibrate CRS.
Sulfated ash (2.4.14) B. Ultraviolet and visible absorption spectrophotometry
Maximwn 0.1 per cent, determined on 1.0 g. (2.2.25).
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2022 Clomifene Citrate 1-625
Test solution (a) Dissolve 0.10 g in methanol R and dilute to Limit:

100.0 mL with the same solvent. Dilute 10.0 mL of this - total: not more than 10 times the area of the peak due to
solution to 100.0 mL with methanol R. clofibrate in the chromatogram obtained with reference
Test soluuon (b) Dilute 10.0 mL of test solution (a) to solution (a) (0.1 per cent).
100.0 mL with methanol R. 4-Chlorophenol
Spectral range 250-350 nm for test solution (a); 220-250 nm Gas chromatography (2.2.28) as described in the test for
for test solution (b). volatile related substances with the following modifications.
Absorption maxima At 280 nm and 288 nm for test Test solution Shake the aqueous layer reserved in the test for
solution (a); at 226 run for test solution (b). volatile related substances with 2 quantities, each of 5 mL, of
Specific absorbanus at the absorption maxima: chlorofonn R and discard the organic layers. Acidify the
- at 226 run: about 460 for test solution (b); aqueous layer by the dropwise addition of hydrochloric addR.
- at 280 om: about 44 for test solution (a); Shake with 3 quantities, each of 3 ml., of chlorofonn R.
- at 288 run: about 31 for test solution (a). Combine the organic layers and dilute to 10.0 mL with
chloroform R.
TESTS
Reference solution Dissolve 0.25 g of chlorophenol R in
chlorofonn R and dilute to 100.0 mL with the same solvent.
1.138 to 1.147. Dilute 1.0 mL of this solution to 100.0 mL with
Refractive index (2.2.6) chloroform R.
1.500 to 1.505. Limit.
Acidity - 4-ch/Qrapheno/: not morethan the area of the peak due to
To 1.0 g add 10 mL of anhydrous emanol Rand 0.1 mL of 4-cWorophenol in the chromatogram obtained with the
phenol red solution R. Not more than 1.0 mL of 0.01 M reference solution (25 ppm).
sodium hydroxide is required to change the colourof the ____________________ 1'1I,,,
indicator.
Volatile related substances
Test solution To 10.0 g of the substance to be examined add Clomifene Citrate
a mixture of 10 mL of dilutesodium hydroxide solution Rand
10 mL of waterR. Shake, separate the lower (organic) layer, (ph. Eur. monograph 0997)
wash with 5 mL of water R and add the washings to the
aqueous layer. Dry the organic layer with anhydrous sodium
sulfate R and use as the test solution. Reserve the aqueous
layer for the test for 4-chlorophenol.
Reference solution (a) Dissolve 0.12 g of the substance to be
(o;,:'H
(OH
examined in chlorofonn R and dilute to 100.0 mL with the
CD,H
same solvent. Dilute 1.0 mL of thissolution to 10.0 mL with
chloroform R. and(l}-isomer
Reference soluticn (b) Dissolve 0.12 g of methyl 2-(4-
chlorophenoxy)-2-methylpropionate CRS in the substance to be C"H,.CINO, 598.1 50-41-9
examinedand dilute to 10.0 mL with the same solvent.
Dilute 1.0 mL of the solution to 10.0 mL with the substance Action and use
to be examined. Dilute 1.0 mL of this solutionto 10.0 mL Estrogen receptor modulator.
with the substance to be examined. Preparation
Column: Clomlfene Tablets
- size: 1= 1.5 m, 0 = 4 mm;
- stationary phase: tilanised diatomaceous earth for gas
I'll'" _
chromatography R (250-420 urn) impregnated with DEFINITION
30 per cent mlm of methylpolysi!oxane R; or silanised 2-[4-[(lEZ)~2-CWoro_l,2-diphenylethen-l-yllphenoxyl-N,N
diatomaceous earthfor gas chromatography R (150-180 urn) diethylethan-l-amine dihydrogen 2-hydroxypropane-I,2,3-
impregnated with 10 per cent mlm of methylpolysiloxane R; tricasboxylate.
Content
Carrier gas nitrogen for chromawgraphy R. - clomifene citrate: 98.0 per cent to 101.0 per cent
Detection Flame ionisation. (anhydrous substance);
Injection 2 ~L. - (Z)-isomer. 30.0 per cent to 50.0 per cent.
System suitability Reference solution (b): CHARACTERS
- peak-to-valley ratio: minimum 4, where Hp = height above Appearance
the baseline of the peak due to methyl 2-(4- Whiteor pale yellow, crystalline powder.
chlorophenoxy)-2-methylpropionate and HfI = height
Solubility
above the baseline of the lowest point of the curve
Slightly soluble in water, sparingly soluble in ethanol
separating this peak from the peak due to clofibrate.
(96 per cent).
IDENTIFICATION
Comparison clomifeue citrate for lD and assayCRS.
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1-626 Clomifene Citrate 2022
B. Dissolve about 5 mg in 5 mL of a mixture of 1 volume of - for impurity A) use the concentration of clomifene citrate
acetic anhydn"de R and 5 volumes of pyridine R, thenheat in a in reference solution (b);
water-bath. A deep red colour is produced. - for impurities other than A, use the concentration of
clomifene citrate in reference solution (c).
TESTS
Prepare the solutions protected from lighl in broom-glass vesseu. Limits:
Ensure minimum exposure of thesolutions to daylight until they - impun'IY A: maximum 1.0 per cent;
are required for chromatography. - impurities CJ D: for each impurity) maximum 0.3 per cent;
Related substances - sum of impurities G and H: maximum 0.15 per cent;
Liquid chromatography (2.2.29). - unspecified impwitie.s: for each impurity, maximum
Testsolution Dissolve 12.5 mg of the substance to be 0.10 per cent;
examined in the mobilephase and dilute to 10.0 mL with - cotal: maximum 2.0 per cent;
the mobile phase. - reporting threshold: 0.05 per cent; disregard the peakdue to
Ref....ce sdiaion (a) Dissolve 12.5 mg of domifene for SYSl<m the citrate ion.
suitabUity CRS (containing impurities A and C) in the mobile Water (2.5.12)
phase and dilute to 10 mL with the mobile phase. Maximum 1.0 per cent) determined on 1.000 g.
ASSAY
Clomifene citrate
Ref....cesolurion (c) Dilute 1.0 mL of reference solution (b) Dissolve 0.500 g in 50 mL of anhydrous acetic acidR. Titrate
to 10.0 mL with the mobile phase. with 0.1 M perchkJric add) determining the end-point
Reference solutWn (d) Dissolve 12.5 mg of c10mifene forpeak potentiometrically (2.2.20).
identi/icarion CRS (containing impurities B and D) in the I mL of 0.1 M perchloric acid is equivalent to 59.81 mg
mobile phase and dilute to 10 mL with the mobile phase. of C,2H"CINOa.
Ref....cesolutWn (e) Dissolve 12.5 mg of clomifene for
(Z)-Isomer
impurities G and H identification CRS in the mobile phase and
Liquid chromatography (2.2.29). Prepare the solutWns protected
dilute to 10 mL with the mobile phase. from light in brown-glass vessels. Ensure minimum exposure of the
Column: solutwns to daylight until they are required for chromatography.
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped burylsi/yl silica gelfor
Mobile phase Mix 400 mL of acetonitrile for chromatography R with the mobile phase.
and 600 mL of water for chromatography R and add 8.0 mL
Reference solution Dissolve 25.0 mg of cIomifene citrate for ID
of dielhylamine R; adjust to pH 6.2 with 1-2 mL of phosphoric
and assayCRS in the mobile phase and dilute to 50.0 mL
acidR. with the mobile phase. Dilute 1.0 mL of the solution to
Flow ral< 1.2 mIlmin. 10.0 mL with the mobile phase.
Detection Spectrophotometer at 233 run. Column:
Injection 10 ~L - size: 1= 0.25 m, 0 = 4.6 mID;
Runtime 4 times the retention time of clomifene. - staMnaryphase: end-capped burylsi/yl silica gelfor
ldendficadon of impurities Use the chromatogram supplied chromatography R (5 um),
with clomifene for system suitability CRS and the chromatogram Mobile phase Mix 0.3 volumes of triethylamine R,
obtained with reference solution (a) to identify the peaks due 45 volumes of waterfor chromatography Rand 55 volumes of
to impurities A and C; use the chromatogram supplied with methanol RI; adjust to pH 2.5 with phosphoric acidR.
clomifene for peak idenrijicatWn CRS and the chromatogram Flow rate 1.0 mUmin.
obtained with reference solution (d) to identify the peaks due Detection Spectrophotometer at 233 om.
to impurities Band D; use the chromatogram supplied with
Injection 50 ~L.
clomifene for impurities G and H idenrijicarion CRS and the
chromatogram obtained with reference solution (e) to identify Run time 1.5 times the retention time of clomifene
the peaks due to impurities G and H. (Z)-isomer. .
Relauve retention With reference to clomifene (retention Idenli/icatWn of peaks Use the chromatogram supplied with
time =about 14 min): citrate = about 0.16; clomifene citrate for ID and (illay CRS and the chromatogram
= =
impurity B about 0.28; impurity D about 0.34; obtained with the reference solution to identify the peaks due
= =
impurity C about 0.5; impurity A about 0.9; impurity G to both isomers of clomifene.
= =
or H about 1.44; impurity G or H about 1.49. Relative retention Withreference to clomifene (Z)-isomer
System suitabibiy Reference solution (a): (retention time = about 13 min): clomifene
- ptak-to-valIey ratio: minimum J5) where Hp = height =
(E)-isomer about 1.2..
above the baseline of the peak due to impurity A and System suitability Reference solution:
H; = heightabove the baseline of the lowest point of the - resolution: minimum 1.5 between the peaks due to
curve separating this peak from the peakdue to clomifene (Z)-isomer and clomlfene (E)-isomer.
clomifene. Measure the area of the peak due to (Z)-isomer in the
Calcularion of percentage contents: chromatograms obtained with the test solution and the
(correction factor multiply the peak area of impurity B by reference solution. Calculate the contentof the (Z)-isomer, as
1.9; a percentage of the total clomifene citrate present) taking into
account the assigned content of (Z)-isomer in clomifene citrate
for ID and (illay CRS.
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2022 Clomipramine Hydrochloride 1-627
CI
STORAGE
IMPURITIES
Specified impurities A, B, C, D, G, H.
Otherdeuaable impurities (the following mbsrances would, if CI

criterion for otherlunspedfied impurities and/or by thegeneral and(l}-isomer
monograph Subsrances for pharmaceutical use (2014). II is
therefore not necessary to idemify these impurities for F. 2- [4- [( IEZ) - [2-chloro-2-(4-ch1orophenyl)-I-phenylethen-
demonstration of compliance. See also5.10. Control oj impurities I-yl]phenoxy]-N,N-diethylethan-I-amine,
in subsronces for pharmaceutical use) E, F.
and(Z}-Isomer
Of' (Z}-Isomer
A. 2-[4-[(IEZ)-I,2-diphenylethen-I-yl]phenoxy]-N,N-
diethylethan-l-amine, GH. 2- [2-chloro-4-[(13)-2-chloro-1,2-diphenylethen-I-yl]
phenoxy]-N,N-diethylethan-I-amine (G. higher-melting-point
isomer; H. lower-melting-point isomer).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Clomipramine Hydrochloride
B. [4-[2-(diethylamino)ethoxy]phenyl]phenyhnethanone,
o O~N-""'CH,
CI
"CH,
andenanUomer . Hel
C. (2RS)-2-[4- [2-(diethylamino)ethoxy]phenyl)-I,2-
diphenylethan-l-one,
351.3 17121-77-6
Action and use

Monoamine reuptake inhibitor; tricyclic antidepressant.
Preparations
Clomipramine Capsules
Clomipramine Prolonged-release Tablets
PIlE" _ _-'--- _
DEFINITION
D. 2,2-bis[4-[2-(d iethyJamino)ethoxy] phenyl]-I,2- 3-(3-Chloro-1 0, II-dihydro-5H-dibenzo[bJ]azepin-5-yl)-N,N-
diphenylethan-l-one, dimethylpropan-l-amine hydrochloride.
Cl
Content
CHARACTERS
Appearance
White or slightly yellow, crystalline powder, slightly
hygroscopic.
Solubllity
Freelysoluble in water and in methylene chloride, soluble in
and(Z}isomer
ethanol (96 per cent).
CI
E. 2-[4-[ (1EZ)-[ I,2-bis(4-chlorophenyl)ethen-I-yl)phenoxy]- IDENTIFICATION
N,N-dlethylethan_l_amine, A. Infrared absorption spectrophotometry (2.2.24).
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1-628 Clomipramine Hydrochloride 2022
Comparison clomipramine hydrochloride CRS. Flow rate 1.5 m1Jmin.

If the spectra obtained show differences, dissolve the Detection Spectrophotometer at 254 run.
substance to be examined and the reference substance Injection 20 ~L.
separately in 2-propanol R, evaporate to dryness and record Relativeretention With reference to clomipramine (retention
new spectra using the residues. time = about 8 min): impurity B = about 0.7;
B. Dissolve about 50 mg in 5 mL of water R and add I mL impurity C = about 0.9; impurity D = about 1.7;
of dilute ammonia Rl. Mix, allow to stand for 5 min and impurity F =about 3.4.
filter. Acidify the filtrate with dilute nilric add R. The solution System suitability Reference solution (c):
gives reaction (a) of chlorides (2.3.1). - resolution: minimum 3.0 between the peaksdue to
TESTS clomipramine and impurity C.
Soludon S Limits:
Dissolve 2.0 g in carbon dioxide-free waterR and dilute to - impun"ty B: not more than the area of the corresponding
20 mL with the same solvent. peak in the chromatogram obtained with reference
Appearance of solution solution (a) (1.0 per cent);
Solution S is clear (2.2.1) and not more intensely coloured - impurities C, D: for each impurity, not more than the area
than reference solution Yj (2.2.2, Method l). of the corresponding peak in the chromatogram obtained
with reference solution (a) (002 per cent);
pH (2.2.3)
- impun'ty F: not more than the area of the corresponding
Related substances solution (a) (0.1 per cent);
Liquid chromatography (2.2.29). Prepani the solUlions - unspecified impun'ties: for each impurity, not more than
immediately befoni use and protected from light. 0.1 times the area of the principal peak in the
Test solution Dissolve 20.0 mg of me substance to be chromatogram obtained with reference solution (b)
examined in a mixture of 25 volumes of mobile phaseBand (0.10 per cent);
75 volumes of mobile phase A and dilute to 10.0 mLwith - sumof unspecified impurities: not more than 0.2 times the
the same mixture of mobile phases. area of the principal peak in the chromatogram obtained
Ref....cesolution (a) Dissolve 22.6 mg of imipramine with reference solution (b) (0.2 per cent);
hydrochloride CRS (impurity B). 4.0 mg of clomipramine - total: not more than the area of the principal peak in the
impurity C CRS, 4.0 mg of clomipramine impurity D CRS and chromatogram obtained with reference solution (b)
2.0 mg of clomipramine imppriiy F CRS in a mixture of (1.0 per cent);
25 volumes of mobile phase Band 75 volumes of mobile - disregard limir. 0.05 times the area of the principal peak in
phase A and dilute to 100.0 mL with the same mixture of the chromatogram obtained with reference solution (b)
mobile phases. Dilute 1.0 mL of this solution to 10.0 mL (0.05 per cent).
with a mixture of 25 volumes of mobile phase Band Loss on drying (2.2.32)
75 volumes of mobile phase A. Maximum 0.5 per cent, determined on 1.000 g by drying in
Reference solution (b) Dilute 1.0 mL of the test solution to an oven at 105 "C.
100.0 mL with a mixture of 25 volumes of mobile phase B Sulfated ash (2.4.14)
and 75 volumes of mobile phase A. Maximum 0.1 per cent, determined on 1.0 g.
Ref....ce solutio" (c) Dissolve 10.0 mg of clomipramine ASSAY
hydrochloride CRS and 3.0 mg of clomipramine impurity C CRS
Dissolve 0.250 g in 50 mL of etha"ot (96 per cent) R and add
in a mixtureof 25 volumes of mobile phase Band
5.0 mL of 0.01 M hydrochloric acid. Carry out a
75.volumes of mobile pbase A and dilute to 20.0 mL with
potentiometric titration (2.2.20), using 0.1 M sodium
the same mixture of mobile phases. Dilute 1.0 mL of this
solution to 10.0 mL with a mixture of25 volumes of mobile
inflexion.
phase Band 75 volumes of mobile phase A.
Column:
C,.,H,.CI,N2 •
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: base-deactivated end-capped cyanopropylsilyl STORAGE
,,1ica gelfor chromatography R (5 um), In an airtight container, protected from light.
- temperature: 30°C. IMPURlTIES
Mobile phase: Specified impurities B, C, D, F.
- mobile phase A: dissolve 1.2 g of sodium dihydrogen Otherdetectable impun'ties (thefollowing substa"ces would, if
phosphate R in water R, add 1.1 mL of .,my/amineR, present at a sufficient level, bedeuaed by one Or other of the tests
adjust to pH 3.0 with phosphoric acidR and dilute to
in the monograph. They ani limited by thegeneral acceptance
1000 mL with water R; criterion for otherlunsprcijied impurities andlor by the general
- mobile phase B: aceumitrile R; monograph Substances for pharmaceutical use (2034). It is
(min) (per cent VIP) (per cent VIJI) demonstration of compliance. See also 5.10. Control of impurities
0-10 7S 25
in substances for pharmaceudcal use) A, E, G.
10·20 75 -+ 65 25 ...... 35
20 - 32 65 35
32 - 34 65 ...... 75 35 -> 25
34 - 44 75 25
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2022 Clonazepam 1-629
cr Cl
N -<:»> N~N"CH3
, ,
CH3 CHJ
A. N-[3-(3-chloro-10, 11-<1ihydr0-5H-dibenzo[bJ]azepin-5-yl} G. 3-chlor0-5-(prop-2-en-l-yl)-10,ll-dihydro-5H-dibenzo[b,

propyl]-N,N',N'-trimelhylpropane-I,3-diamine, j)azepine.
--- ""EII
••••
Clonazepam ••* *
* •••
B. 3-(10,11-dihydro-5H-dibenzo[bJ]azepin-5-yl)-N,N-
o,N~~
I ~Jo
dimethylpropan-l-amine (imipramine),
cr
1 "" Cl
---"
315.7 1622-61-3
Action and use

C. 3-(3-chloro-5H-dibenzo[bJ]azepin-5-yJ}-N,N- Benzodiazepine.
dimethylpropan-l-amine, Preparations
cr
Clonazepam Injection
Clonazepam Oral Suspension
Clonazepam Tablers
""Ell _ _ ~ _
DEFINITION
5-(2-Chlorophenyl)-7-nltro-I ,3-dihydro-2H-1 ,4-
benzodiazepin-2-one.
D. 3-(3,7-dichloro-10,ll-dihydr0-5H-dibenzo[bJ]azepin-5- Content
yl}-N,N-dimethylpropan-l-amine, 99.0 per cent to 101.0 per cent (dried substance).
CHARACTERS
Appearance
Slightly yellowish, crystalline powder.
Solubility
Practically insoluble in water, slightly soluble in alcohol and
in methanol.
mp
E. 10, ll-dihydro-5H-dibenzo[bJ]azepine (iminodibenzyl), About 239 'C.
IDENTIFlCATIO!'l
Cl
Comparison Ph. Bur: reference spectrum of donazepam.
TESTS
Related substances
Liquid chromatography (2.2.29). Carry out the rest protected
from light and prepare the solutions immediately before use.
Soioen: mixture tetrahydro/uran R, methanol R, water R
F. 3-<:IlIoro-l 0, ll-dihydr0-5H-dibenzo[bJ] azepine, (10:42:48 VIVJII).
solvent. Dilute 1.0 mL to 10.0 mL with the solvent mixture.
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1-630 Clonidine Hydrochloride 2022
~
Reference solution (a) Dilute 1.0 mL of the test solution to N H'
100.0 mL with the solvent mixture. Dilute 1.0 mL of the

solution to 10.0 mL with the solventmixture. o,N
"'" I 0
Reference solution (b) Dissolve 5 mg of the substance to be "" CI

examined and 5 mg ofjlunitrazepam R in the solvent mixture 1#
and dilute to 100.0 mL with the solventmixture.
Reference solution (c) Dissolve 1.0 mg of donazepam A. (2-amino-5-nitrophenyl)(2-ebloropbenyl)methanone,
impunty B CRS in the solvent mixture and dilute to 20.0 mL
~
with the solvent mixture. Dilute 1.0 mL of the solution to ~ 0
Column: o,N "'" I # NH,
- size: 1-= 0.15 m, 0 ;;;; 4.6 D1ITI,
- stationary phase: end-copped octy!si/yl silica gelfur "" cr
chromarography R (5 urn). 1#
Mobile phase Mix 10 volumes of tetrahydrofuran R,
42 volumes of methanol Rand 48 volumes of a 6.6 gIL B. 3-amino-4-(2-chlorophenyl)-6-nitroquinolin-2(1H)-one.
solution of ammonium phosphate R previously adjusted to _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r
pH 8.0 with a 40 gIL solution of sodium hydroxide R or dilute
phosphotic acidR.
Flowrate 1.0 mUmin.
Detection Spectrophotometer at 254 nm, Clonidine Hydrochloride
Injection I0 ~L.
Run lime 3 times the retention time of c1onazepam.
Relative retention With reference to clonazepam (retention
• Hct
impurity A = about 2.4-
flunitrazepam and to clonazepam, 266.6 4205-91-8
limits:
- impurity A: not more than the area of the principal peak Action and use
in the chromatogram obtained with reference solution (a) Alpha2-adrenoceptor agonist; treatment of hypertension.
(0.1 per cent), Preparations
- impun'ty B: not more than the area of the principal peak in Clonidine Injection
the chromatogram obtained with reference solution (c) Clonidine Tablets
(0.1 per cent)
PhE<r _
- attV other impurity: for each impurity, not more than the
area of the principal peak in the chromatogram obtained DEFINITION
with reference solution (a) (0.1 per cent), 2,6-Dicbloro-N-(intidazolidin-2-ylidene)aniline hydrocbloride.
- total: not more than twice the area of the principal peakin
the chromatogram obtained with reference solution (a) Content
(0.2 per cent), 98.5 per cent to 101.0 per cent (dried substance).
- disregard [ittn!: 0.5 timesthe area of the principal peak in CHARACTERS
the chromatogram obtained withreference solution (a) Appearance
(0.05 per cent). White or almost while, crystalline powder.
Loss on drying (2.2.32) Solubility
Maximum 0.5 per cent, determined on 1.000 g by drying in Soluble in water and in anhydrous ethanol.
an oven at lOS °C for 4 h.
IDENTIFICATION
Sulfated ash (2.4.14) First identification: B, D.
Second identification: A J CJ D.
ASSAY A. Ultraviolet and visible absorption spectrophotometry
Dissolve 0.275 g in 50 mL of acetic anhydride R. Titrate with (2.2.25).
0.1 M perchlotic acid, determining the end-point Test solution Dissolve 30.0 mg in 0.01 M hydrochloric acid
I mL of 0.1 M perchioric acid is equivalent to 31.57 mg Spearal rallge 245-350 nm.
of ClsHlOC1N303.
Absorption maxima At 272 nm and 279 nm.
STORAGE Point of inflexioN At 265 nm.
Specific absatixmce at the absorption maxima:
IMPURITIES - at 272 om: about 18;
S]Je<ified impurities A, B. - at 279 nm: about 16.
Comparison clonidine hydrochloride CRS.
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2022 Clonidine Hydrochloride 1-631
C. Thin-layer chromatography (2.2.27). Flow rate 1.5 mUmlri.

Test solution Dissolve 5 mg of the substance to be examined Detection Spectrophotometer at 210 nm.
in methanol R and dilute to 5 mL with the same solvent. Injection 5 ~L.
Reference solution Dissolve 5 mg of clonidine System suitability Reference solution (b):
hydrochloride CRS in methanol R and dilute to 5 mL with the - resolution: minimum 5.0 between the peaks due to
same solvent. clonidine and impurity B.
Plate TLC silica gel G plate R. Limits:
MobIle phase glacial acetic acid R, butanol R, water R - unspecified impun·Ues;, for each impurity, not more than the
(10:40:50 VIV/JI); allow to separate, filter the upper layer area of the principal peak in the chromatogram obtained
and use the filtrate. with reference solution (a) (0.10 per cent);
Application I 0 ~L. - total: not more than twice the area of the principal peak in
Development Over 2/3 of the plate. the chromatogram obtained with reference solution (a)
(0.2 per cent);
Drying In air. - disregard limit: 0.5 times the area of the principal peak in
Detection Spray with potassium iodobismuthate solution R2. the chromatogram obtained with reference solution (a)
Allow to dry in airfor 1 h. Sprayagainwith potassium (0.05 per cent).
iodobismuthate solution R2 and then immediately spray with a
50 gIL solution of sodium nitrite R.
Results The principal spot in the chromatogram obtained an oven at 105°C.
the principal SPO[ in the chromatogram obtained with the
reference solution. Maximum 0.1 per cent, determined on 1.0 g.
D. It gives reaction (a) of chlorides (2.3.1). ASSAY
Dissolve 0.200 g in 70 mL of ethanol (96 per renO R. Titrate
TESTS with 0.1 M ethanolic sodium hydroxide determining the
Solution S end-point potentiometrically (2.2.20).
Dissolve 1.25 g in carbon dioxide-free water Rand dilute [0
25 mL with the same solvent, I mL of 0.1 lW ethanofic sodium hydroxide is equivalent to
26.66 mg of C.HlOCl,N,.
Solution S is clear (2.2.1) and not more intensely coloured IMPURITIES
than reference solution Y7 (2.2.2, MethodII). Otherdaeaable impurities (thefollowing substances would, if
present Ql. a sufficient level, be detected by oneor other of the tests
pH (2.2.3)
Related substances monograph Substances for phannaceutical use (2034). It is
liquid chromatography (2.2.29). therefore not necessary 10 identify these impurities for
Test solution Dissolve 50 mg of the substance to be demonstration of compliance. See also 5.10. Control of impurities
examined in mobile phase A and dilute to 50 mL with in substanus for pharmaceutical use) A, B, C.
mobile phase A.
Reference solution (b) Dissolve 5 mg of clonidine
impurity B CRS in 2 mL of acetonini/e Rand dilute [0 5 mL
with mobile phase A. To 1 mL of this solution, add I mL of A. l-acetylimidazoildin-2-one,
the test solution and dilute to 10 mL with mobilephase A.
Column:
- size: 1= 0.15 m, 0 = 3.0 mm;
- stationary phase: propylsilyl silica gelfor chromawgraphy R
(5 pm),
Mobile phase: B. l-acetyl-2-[(2,6-dichlorophenyl)amino]-4,5-dihydro-IH-
- mobile phaseA: dissolve 4 g of potassium dihydrogen imidazole,
phosphate R in 1000 mL of water for chromatography R,
and adjust to pH 4.0 with phosphoric acid R;
- mobile phase B: mobile phase A, acetonitrile R1
(25:75 V/JI);
Thn. MobUe phase A MobUe phase B

(min) (per cent VIP) (per cent VIJ1
C. 2,6-dichloroaniline.
0 90 10
o~ 15 90 --J 30 10 ..... 70 PhE"
15 - 15.1 30 --J 90 70 --J 10
15.1 - 20 90 10
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1-632 Clopamide 2022

Clopamide - mobile phase C: waterR, tetrahydrofuran for
(ph. Bur. monograph 1747) chromarography R (20:80 V/II); this mobile phase aUows
adequate rinsing of the system;
Time Mobile phase A Mobile phase B Mobile phase C

(min) (per cent VIP) (per cent VIJI) (per cent VIJI)
0·35 95 -> 75 5 -> 25 0
35 - 45 75 --0 35 25 --0 65 0
45 - 50 35 -> 30 65 -> 0 0->70
50 - 60 30 0 70
345.8 636-54-4
Action and use Flow rale 0.4 mUmin.

Thiazide-like diuretic. Deeaion Spectrophotometer at 235 run.
1'1181 _ Injection I0 ~.
DEFINlTION with clopamide for system suitabl7ity CRS and the
4-Chloro-N-[(2RS,6SR)-2,6-dimethylpiperidin-I-yl)-3- chromatogram obtained with reference solution (a) to
sulfamoylbenzamide. identify the peaks due to impurities B, C and H.
Content RelatifJe retention With reference to clopamide (retention
99.0 per cent tn 101.0 per cent (dried substance). time = about 33 min): impurity C = about 0.8;
PRODUCTION
impurity H = about 1.2; impurity B = about 1.4.
The production method is evaluated to determine the System suitability Reference solution (a):
potential for formation of an N-nitroso compound (cir-2,6- - resohuion: minimum 3 between the peaks due to
dimethyl-l-nitrosopiperidine). Where necessary, me impurity C and clopamide.
production method is validated to demonstrate that the Limits:
N-nitroso compound is absent in the final product. - correction factors: for the calculation of content, multiply
CHARACTERS
corresponding correction factor: impurity B = 0.5;
Appearance
White or almost white, hygroscopic, crystalline powder.
impurity H =0.4;
- impurities B, C, H: for each impurity, not more than twice
Solublllty the area of the principal peak in the chromatogram
Slightly soluble in water and in anhydrous ethanol, sparingly obtained with reference solution (b) (0.2 per cent);
soluble in methanol. - unspecified impurities: for each impurity, not more than the
It shows polymorphism (5.9). area of the principal peak in the chromatogram obtained
IDENTIFICATION
- wud: not more than 10 times the area of theprincipal
Comparison clopamide CRS. solution (b) (1.0 per cent);
If the spectra obtained in the solid stateshow differences, - disregard limir. 0.5 times the area of the principal peak in
dissolve the substance to he examined and the reference the chromatogram obtained with reference solution (b)
substance separately in the minimum volume of methanol R, (0.05 per cent).
evaporate to dryness on a water-bath andrecord new spectra Loss on drying (2.2.32)
usingthe residues. Maximum 2.5 per cent, detennined on 1.000 g by drying in
TESTS an oven at 105 "C.
Test solution Dissolve 100 mg of the substance to be ASSAY
examined in methanol R and dilute to 10.0 mL with the same Dissolve 0.280 g in 70 mL of anhydrous acetic add R. Titrate
solvent. with 0.1 M perchloric acid, determining the end-point
Reference solution (a) Dissolve 10 mg of clopamide for syuem potentiometrically (2.2.20).
suitability CRS (containing impurities B, C and H) iIi 1.0 mL
of methanol R.
of C J4H,oCIN,O,S.
100.0 mL with methanol R. Dilute 2.0 mL of this solution to STORAGE
40.0 mL with methanol R. In an airtight container, protected from light.
Column: IMPURITIES
- size: 1= 0.15 m, (2) = 4.6 mrn; Specified impurities B, C, H.
- stationary phase: end-capped octy/silyl silica gelfor Otherdetectable impurities (thefollowing substances would, if
chromarography R (5 urn). present at a sufficient level, be detected by oneor other of the tests
Mobile phase: in the monograph. They are limited by the general a«eptance
- mobile phase A: dissolve 1.0 g of ammonium acetate R in criterion for other/unst=ified impuniies and/or by thegeneral
950 mL of waterR, adjust to pH 2.0 with phosphoric monograph Substances for pharmaceuti<:o1 use (2034). It is
acidR and dilute to 1000 mL with waterR; therefore not necessary to identify these impurities for
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2022 Clopidogrel Besilate 1-633
demonstration of compliance. See also 5.10. Control of impurities PRODUCTION

in substances for pharmaceutical use) A, G. It is considered that alkyl benzenesulfonate esters are
genoroxic and arepotential impurities in clopidogrel besilate.
,slY .
H,C
The manufacturing process should be developed taking into
\\ 0
0 Ii OHXI"
N
and enantiomet together with considerations of the quality of starting
H,N I ~ ..
R H CH, materials, process capability and validation. The general
CI method 2.5.41. Methyl, ethyland isopropyl beneenesulfonau in
active substances is not suitable for c1opidogrel besilate since it
A. 4-cWoro-N-[(2RS,6RS)-2,6-dimethylpiperidin-l-yl)-3- was observed thatmethyl benzenesulfonate was obtained
sulfamoylbenzamide (trans-dopamide), during the gas chromatography analysis as an artefact
originating from degradation. Another suitable and validated
~Co,H method should be used. The content of each alkyl
benzenesulfonate is not more than 3 ppm.
Cl.AvJ
CHARACTERS
B. 4-chlorohenzoic acid, Appearance
o 0
Solublllty
H,N
"'" I
,SOCo,H Practically insoluble in water, freely soluble in anhydrous
CI R
IDENTIFICATION
C. 4-chloro-3-sulfamoylbenzoic acid, Carry out either testsAJ BJ D or testsBJ CJ D.
A. Specific optical rotation (2.2.7): + 47.0 to + 51.0
H,C
. 00 OHt)' Dissolve 0.250 g in methanol R and dilute to 25.0 mL with
H,N ~'iXi'''''
I ~ ,N
and enanliomer
the same solvent.
CI
Comporison clopidogrel besilate CRS.
G.4-cWoro-N-[(2RS)-2-methylpiperidin-l-yl]-3- C. Enantiomeric purity (see Tests).
sulfamoylbenzamide, D. Liquld chromatography (2.2.29) as described in the lest
for related substances with the following modification.
Injection Test solution and reference solution (d).
Results:
- thepeak due to besllate in the chromatogram obtained
with the test solution is similar in retention time to the
corresponding peak in the chromatogram obtained
H.4-cWoro-3-[(E)-[(dimethylamino)methylene]sulfamoyl]-N- with reference solution (d);
[(2RS,6SKJ-2,6-<1imethylpiperidin-I-yl)benzamide. - the ratio of the area of the peak due to the besilate to
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE" the area of the peak due to c1opidogrel in the
chromatogram obtained with the test solution is
minimum 0.1.
TESTS
Clopidogrel Besilate ***** Enantiomeric purity
** *
*** *
Liquld chromatography (2.2.29).
(ph. Bur. monograph 2790) Testsolution (a) Dissolve 46.0 mg of the substance to be
examined in 10.0 mL of anhydrous ethanol R and dilute to
38
CI \,. 20.0 mL with heptane R.
rr~H\
Test solution (b) Dilute 1.0 mL oftest solution (a) to
OCH, 10.0 mL with heptane R.
\-V 0
Reference solution (a) Dissolve 10 mg of dopidogrel for system
suitabilisy CRS (containing impnrities B and C) in 2.5 mL of
anhydrous ethanol R and dilute to 5.0 mL with heptane R.
C"H"CINO,S, 480.0 744256-69-7
Reference solution (b) Dilute 1.0 mL of test solution (a) to
Action and use 100.0 mL with heptane R. Dilute 1.5 mL of this solution to
Inhibitor of ADP-mediated platelet aggregation. 10.0 mL with heptane R.
Reference solution (c) Dissolve 34.0 mg of dopidogrel
PIlE" _
hydrochloride CRS in 10.0 mL of anhydrous ethanol Rand
DEFINlllON dilute to 20.0 mL with heptane R. Dilute 1.0 mL of the
Methyl (2S)-2-(2-chlorophenyl)-2-[6,7-dihydrothieno[3,2- solution to 10.0 mL with heptane R.
c]pyridin-5(4H)-yI]aceta te benzenesulfonare. Column:
Content - size: 1= 0.25 IDJ 0 = 4.6 mm;
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1-634 Clopidogrel Besilate 2022
- stationary phase: cellulose derivative of silica gelfor chiral lnjution 10 ~ of tile test solution and reference
separation R (10 urn). solutions (b) and (c).
Mobile phase anhydrous erhanol R, heptane R (15:85 VIV). Identification of inrpun"eies Use the chromatogram supplied
Flow rate 0.8 mUmin. with dopidogrel for system sm'tabih'ty CRS and the
Detection Spectrophotometer at 220 run. chromatogram obtained with reference solution (b) to
Injection 10 J1L of test solution (a) and reference
ReJat1ve retention With reference to c1opidogrel (retention
time = about 27 min): besilate = about 0.05j
Run time 1.25 times the retention time of clopidogrel. impurity A =about 0.4; impurity B =about 1.1.
Identification of impurities Use the chromatogram supplied System suitability Reference solution (b):
with c/qpidogrel for system suitability CRS and the - peak-to"!/alley ratio: minimum I 0, where Hp = height
chromatogram obtained with reference solution (a) to above the baseline of the peak due to impurity Band
identify the peaks due to impurities B and C. H l1 = height above the baseline of the lowest point of the
Relative retention With reference to clopidogrel (retention curveseparating this peak from the peakdue to
time = about 18 min): impurity C = about 0.6; clopldogrel.
=
impurity B about 0.7. Calculation of percentage contents:
System suitability Reference solution (a): - for each impurity, use the concentration of clopidogrel
- resolution: minimum 2.0 between the peaks due to besilate in reference solution (c).
impurities C and B. Limits:
Calculation of percentage content: - impurities A, B: for each impurity, maximum
-. for impurity C, use the concentration of c1opidogrel 0.15 per cent;
besilate in reference solution (b). - unspecified impurities: for each impurity, maximum
Limit: 0.10 per cent;
- impun"ty C: maximwn 0.15 per cent. - total: maximum 0.4 per cent;
Related substances - reporting threshold: 0.05 per cent; disregard the peak due to
Liquid chromatography (2.2.29). the besilate.
Solvent mixture Mobile phase A, acetonitrile RI (40:60 VIV). Water (2.5.12)
examined in the solvent mixture and dilute to 10.0 mL with Sulfuted ash (2.4.14)
the solvent mixture. Maximum 0.1 per cent, determined on 1.0 g.
Reference solution (a) Dissolve 5 mg of dopidogrel ASSAY
impurity A CRS in the solvent mixture and dilute to 25.0 rnL Liquid chromatography (2.2.29) as described in the test for
with the solvent mixture. enantiomeric purity with the following modification.
Reference solution (b) Dissolve 32 mg of dopidogrel for system Inj'ection Test solution (b) and reference solution (c).
suitability CRS (containing impurities B and C) in the solvent Calculate the percentage content of Y2H22ClN05S2 taking
mixture, add 0.5 mL of reference solution (a) and dilute to into account the assigned content of dopidogrel
5.0 rnL with the solvent mixture. hydrochloride CRS, and a conversion factor of 1.34.
Reference solUlion (c) Dilute 1.0 rnL of the test solution to
100.0 rnL with the solvent mixture. Dilute 1.0 rnL of this
STORAGE
Reference solution (d) Dissolve 25 mg of sodinm IMPURITIES
bmeenesulfonote R in the solventmixture and dilute to Specified impnrities A, B, C.
10.0 rnL with the solvent mixture. Otherdete<table impurities (rhe following substances would, if
Column: present at a sufficient level, be dete<ted by oneor other of the tests
- size: 1= 0.15 m, 0 = 3.9 mm, in the monograph. They a.. limited by thegeneral acceptance
- stationary phase: end-capped octad«y/siJy1 silica gelfor criterion for orherlunspecified impuniies andlor by thegeneral
chromatography R (5 urn); monograph Substances for pharmaceuti<aI use (2034). It is
- temperatu..: 30 °C. therefore not necessary to identify these impurities for
Mobile phase: demonstration of compliance. See also 5.10. Control of impurities
- mobile phaseA: mix 5 volumes of methanol R2 and in substances for pharmaceutical use) ,D, E, F, G.
95 volumes of a 0.96 gIL solution of sodium
pentanesulfonate monohydrate R adjusted to pH 2.5 with
phospho';'; acid R;
- mobile phase B: methanol R2, acetonitrile RI (5:95 VIV);

(min) (per cent VIP) (percent VIP)
0-3 89.5 10.5 A. (2S)-2-(2-ch1orophenyl)-2-[6,7-dihydrothieno[3,2-
3 - 48 89.5 ...... 31.5 10.5 ...... 68.5 c]pyridin-5(4H)-yl]acetic acid,
48 - 68 31.5 68.5
Flow rate I. 0 mUmin.

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2022 Clopidogrel Hydrochloride 1-635
""E" _
DEFINITION
Methyl (2S)-2-(2-cWorophenyl)-2-[6,7-dihydrothieno[3,2-
c]pyrldin-5(4H)-yl]acetate hydrochloride.
Content
B. methyl (2S)-2-(2-cWorophenyl)-2-[4,7-dihydrothieno[2,3- CHARACTERS
c]pyrid in-6 (5H)-yl] acetate, Appearance
White or yellowishpowder.
Solubility
Practically insoluble in water, very soluble in anhydrous
IDENTIFICATION
Carry out either tests A, B, D or tests B, C, D.
C. methyl (2R)-2-(2-cWorophenyl)-2-[6,7-dihydrothieno[3,2- A. Specific optical rotation (2.2.7): + 65.0 to + 69.0
c]pyridin-5( 4H) -yl]acetate, (anhydrous substance).
the same solvent.
Comparison c1opidogrel hydrochlonae CRS.
TESTS
D. (lR)-I-(2-cWorophenyl)-2-methoxy-2-oxoethyl (2S)-2-(2- Enantiomerlc purity
chlorophenyl) -2-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)- Liquid chromatography (2.2.29).
yl)acerate,
examinedin 10.0 mL of anhydrous ethanol R and dilute to
andenantiomer
Testsolution (b) Dilute 1.0 mL of test solution (a> to
Reference solution (a) Dissolve 10 mg of chpichgre1 for syuem
suitability CRS (containing impurities B and C) in 2.5 mL of
E. (2RS)-2-(2-chlorophenyl)-2-[6,7-dihydrothieno[3,2- anhydrous ethanolR and dilute to 5.0 mL with heptane R.
c]pyridin -5(4H)-yl] acetamide, Reference solution (b) Dilute 1.0 mL of test solution (a) to
100.0 mL with heptane R. Dilute 1.5 mL of this solution to
Reference solution (c) Dissolve 34.0 mg of c1opidogre1
hydrochkJride CRS in 10.0 mL of anhydrous ethanol Rand
dilute to 20.0 mL with heptane R. Dilute 1.0 mL of the
solution to 10.0 mL with heptane R.
F. methyl (2S)-2-(2-cWorophenyl)-2-[[2-(thiophen-2-yl) Column:

- size: 1= 0.25 m, 0 = 4.6 mm;
ethyl]amino]acetate)
- stationary phase: cellulose derivauve of silica gelfor chiral
G. unknown structure. separation R (10 /lID).
___________________ ""E" Mobile phase anhydrous ethanol R, heptane R (15:85 VIV).
Clopidogrel Hydrochloride **** Inj«uCm 10 ilL of test solution (a) and reference
** * solutions (a) and (b).
(ph. Eur. monograph 2791) ***** Run time 1.25 times the retention time of clopidogrel,
Identification of impuriues Use the chromatogram supplied
with dopichgre1 for system suitabi7ity CRS and the
chromatogram obtainedwith reference solution (a) to
• HCl identify the peaks due to impurities Band C.
Relative retention With reference to clopidogrel (retention
=
358.3 120202-65-5 System suitability Reference solution (a):
Action and use - resolution: minimwn 2.0 between the peaks due to
Inhibitor of ADP-mediated platelet aggregation. impurities C and B.
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1-636 CJopidogrel Hydrochloride 2022
Calculation ofpercentage content: - unspecified impurities: for each impurity, maximum

- for impurity C, use the concentration of clopidogrel 0.10 per cent;
hydrochloride in reference solution (b). - total: maximum 0.4 per cent;
Limit: - reponing threshold: 0.05 per cent.
- impurity C: maximwn 0.15 per cent. Water (2.5.12)
Liquid chromatography (2.2.29). Replace the solvent after each titration.
Solvent mixture Mobile phase A, aceumilri/e Rl (40:60 V/II). Sulfated ash (2.4.14)
Test solution Dissolve 55 mg of the substance to be Maximum 0.1 per cent, determined on 1.0 g.
examined in the solvent mixture and dilute to 10.0 mL with ASSAY
thesolvent mixture. Liquid chromatography (2.2.29) as described in the test for
Reference solution (a) Dissolve 5 mg of clopidogrel enantiomeric purity with the following modification.
impun"Cy A CRS in the solvent mixture and dilute to 25.0 mL Injection Test solution (b) and reference solution (c).
Calculate the percentage content of C,,;H17CI,NO,S taking
Reference solution (b) Dissolve 32 mg of clopidogre1 for system into account the assigned content of dopidogrel
suitability CRS (containing impurities Band C) in the solvent hydrochloride CRS.
mixture, add 0.5 mL of reference solution (a) and dilute to
5.0 mL with the solvent mixture. STORAGE
100.0 mL with the solvent mixture. Dilute 1.0 mL of this IMPURITIES
solution to 10.0 mL with the solvent mixture. Specified impumies A, B~ C.
Column: Otherdtte<table impuriti<s (thefollowing substances uotdd, if
- size: 1:;;; 0.15 m, 0 ::::; 3.9 mm; present at a sufficient level, be detected fry one or other of the tests
- stationary phase: end-eapped oaade<ylsilyl silica gelfor in the monograph. Theyart limited by thegeneral acceptance
chromatography R (5 um); aitesion for otherlunspecrfied impun·tie.s and/or by thegeneml
- temperature: 30 DC. monograph Substances for pharmaceutical use (2034). It is
Mobile phase: therefore nornecessary 10 identify these impurities for
- mobae phaseA: mix 5 volumes of methanol R2 and demonstration of compliance. See also 5.10. Control of impurities
95 volumes of a 0.96 gIL solution of sodium in substances for pharmaceutical use) DJ E, F, G.
pentanesulfonate monohydrate R adjusted to pH 2.5 with
phosphoric acidR;
- mobile phaseB: methanol R2, aceumilrile Rl (5:95 VII');
Tim. Mobile phose A Moblle phase B

(min) (per cent 1'/11) (per cent VIJI)
0-3 89.5 10.5
3 - 48 89.5 -> 31.5 10.5 -> 68.5
A. (2S)-2-(2-cWorophenyl)-2-[6,7-dihydtothieno[3,2-
48-68 31.5 68.5
cJpyridin-5(4H)-yIJacetic acid,
Flow rau 1.0 mUmin.
& C~~H'
Injection 10 J1L of the test solution and reference
Identification of impurities Use the chromatogram supplied '" 0
with clopUWgrel for system suitability CRS and the S
B. methyl (2S)-2-(2-cWorophenyl)-2-[4,7-dihydsothieno[2,3-
Relativeretention With reference to c1opidogrel (retention c)pyridin-6(5H)-yl)acetate,
- penk-w-valley ratio: minimum 10, where Hp =height
H; = height above the baselineof the lowest point of the
curve separating this peak from the peakdue to
c1opidogrel.
C. methyl (2R)-2-(2-cWorophenyl)-2-[6,7-dihydsothieno[3,2-
Calculation of percentage contents: cj pyridin-5(4H) -yllacetate,
- for each impurity, use the concentration of clopidogrel
hydrochloride in reference solution (c).
Limits:
- impuritres A, B: for each impurity, maximum
0.15 per cent;
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2022 Clopidogrel Hydrogen Sulfate 1-637
A. Specific optical rotation (2.2.7): + 54.0 to + 58.0

the same solvent.
Comparison cbJpidogrel hydrogen sulfate CRS.
D. (IR}-I-(2-chlorophenyJ}-2-methoxy-2-oxoethyJ (2S}-2-(2-
chlorophenyl}-2-[6,7-dihydrothieno[3,2-<]pyridin-5(4H)-
separately in anhydrous ethanol R, evaporate (0 dryness and
yl]acetate,
record new spectra using the residues (the substance may
stick to the surface of the recipient used).
O:CI . H
D. Ir gives reaction (a> of sulfates (2.3.I}.
rf~XyNH' andenanUomar
TESTS
\AJ 0 Appearance of solution
E. (2RS}-2-(2-chlorophenyl)-2-[6,7-dihydrothieno[3,2- than reference solution Y6 (2.2.2-, Method I).
c]pyrid in-5(4H)-yl] acetamide, Dissolve 1.0 g in methanol R and dilute to 20.0 mL with the
same solvent.
Enantiomeric purity
procedure.
Test solution Dissolve 0.1 g of the substance to be examined
in 25.0 mL of anhydrous ethanol R and dilute to 50.0 mL
with heptane R.
F. methyl (2S}-2-(2-chlorophenyl)-2-[[2-(thiophen-2-yl) Reference solution Dissolve 10 mg of clapidogrel for system
ethyl]amino]aceta te, suitability CRS (containing impurities B and C) in 2.5 mL of
G. unknown structure. anhydrons ethanol R and dilute to 5 mL with heptane R.
~~~ -'---_ _~ ~ Phf" Column:
- size: 1 = 0.25 rn, {2) =: 4.6 mrn;
- stationary phase: cellulose derivative of silica gelfor chiral
.............. separation R (10 urn).

Clopidogrel Hydrogen Sulfate
.......
Mobile phase anhydrous ethanol R, heptane R (15:85 VIII).
Flow rate 0.8 mUmin .
CI Injection 10 ~L.
:i?-DCH,
Run time 1.25 times the retention time of clopidogrel.
rf~H.. Identification of impurities Use the chromatogram supplied
with cIopidogrel for system suitability CRS and the
\AJ 0 chromatogram obtained with the reference solution to
identify the peaks due to impurities Band C.
419.9 120202-66-6 Relative retention With reference to clopidogrel (retention
= =
time about 18 min): impurity C about 0.6;
Action and use =
Inhibitor of ADP-mediated platelet aggregation. System suu!lbility Reference solution:
Phf" _ _ ~~~~ ~ ~_
impurities C and Br
DEFINITION - signal-to-noise ratio: minimum 20 for the peak due to
Methyl (2 S)-(2-chlorophenyl) [6,7-dihydrothieno[3 ,2-e] impurity C.
pyridin-5(4H)-yl]acetate sulfate. Limit:
Content - impuniy C: maximum 0.5 per cent.
99.0 per cent to 101.0 per cent (anhydrous substance). Related substances
CHARACTERS Liquid chromatography (2.2.29).
Appearance Solvent mIXture Mobile phase A, acetonitrile R (40:60 VIII).
White or almost white powder. Test solution Dissolve 65 mg of the substance to be
Solubility examined in the solvent mixture and dilute to 10.0 mL with
Freely soluble in water and in methanol, practically insoluble the solvent mixture.
in cyclohexane. Reference solution (a) Dissolve 5 mg of c/opi<Iogrel
It shows polymorphism (5.9). impurity A CRS in the solvent mixture and dilute to 25 mL
IDENTIFICATION with the solvent mixture.
Carry out either tests A, B, D or tests B, C, D.
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1-638 Clopidogrel Hydrogen Sulfate 2022
Reference solution (b) Dissolve 32 mg of clopidogre/ for sysrem ASSAY

suitability GRS (containing impurities B and C) in the solvent Dissolve 0"160 g in a mixture of 10 mL of acetone R) JO mL
mixture, add 0.5 mL of reference solution (a) and dilute to of methanol Rand 30 mL of water R. Titrate with 0.1 J\1
5 mL with the solvent mixture. sodium hydroxide, determining the end-point
Reference solution (c) Dilute 1.0 mL of the test solution to potentiometrically (2.2.20). A precipitate may be formed
100.0 mL with the solvent mixture. Dilute 1.0 mL of this duringthe titration"
solution to 10.0 mL with the solventmixture. I mL of 0.1 M sodium hydroxide is equivalent to 20.99 mg of
Column: C 16H1SCINO.S,.
- size: I::: 0.15 m, 0 ::: 3.9 mm; STORAGE
- sratlimary phase: end-capped octadecy/silyl SIlica gelfor Protected from light.
- tempera..re: 30 "C. IMPURITIES
Specifi.dimpurities A, B, C.
MoM. phase:
- mobile phase A: mix 5 volumes of methanol RJ and Otherdetectable impurities (the following subsMnces would, if
95 volumes of a 0.96 gIL solution of sodium present at a sufficient level, be detected by oneor other of the teus
pentanesulfonme monohydrate R adjusted to pH 2.5 with in the monograph. They am limitedby thegeneral acceptance
phosphoric acid R; aiunon for other/unspecified impurities and/or by thegeneral
- mobile phaseB: methanol RJ, acetonitrile for monograph Substances for pharmaceutical use (2034). I, is
chromatography R (5:95 VII'); there/ore not neussary to identify these impurities for
Time MobUe phase A MobUe phase B in substances for pharmaceutical use) D.
(min) (per cent VA? (per cent I'll?
0·3 89.5 105
3 - 48 89.5 ---> 31.5 10.5 ---> 68.5
48 - 68 31.5 68.5

Detection Spectrophotometer at 220 nm. A. (2S)-(2-cWorophenyl) [6,7-dihydrothieno[3,2-e]pyridin-5
Injection 10 J.lL of the test solution and reference (4H)-ylJacetic acid,
Identification of impun"ties Use the chromatogram supplied
with cIopidogre/ for system suiU1hiJity CRS and the
Relative retention With reference to clopidogrel (retention
=
time about 25 min): impurity A about 0.4; = B. methyl (2S)-(2-cWorophenyl)[4,7-dihydrothieno[2,3-e]
pyridin-6(5H)-yl]acetate,
- peak-ro-valky ratio: minimum 10) where Hp :;;: height
CUIVe separating this peak from the peak due to
clopidogrel.
Limits:
- impun"ty B: not more than 3 times the area of the C. methyl (2R)-(2-chlorophenyl) [6,7-dihydrothieno[3,2-c]
principal peak in the chromatogram obtained with pyridin-5(4H)-yl] acetate,
- impun"ty A: not more than twice the area of the principal
- unspedjied impurities: for each impurity, not more than the
in the chromatogram obtained with reference solution (c) D. methyl (2R)-(2-cWorophenyl)[(2S)-(2-cWorophenyl)[6,7-
(0.5 per cent); dihydrothieno[3,2-c]pyridin-5(4H)-yl]acetyloxy]acetate.
- disregard limit: 0.5 times the area of the principal peak in _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
the chromatogram obtainedwith reference solution (c)
(0.05 per cent).
Water (2.5.12)
Replacethe solvent aftereach titration"
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2022 Clotrimazole 1-639
Clotrimazole *** Test solution Dissolve 50.0 mg of the substance to be

*** *** examined in acetonitrile R and dilute to 50.0 mL with the
(Ph. Eur. monograph 0757) *** same solvent.
Reference solurion (aJ Dilute 1.0 mL of the test solution to
100.0 mL with acet<mitrik R. Dilute 1.0 mL of this solution
9?0
CI (N
L NIJ
to 10.0 mL with aceton/oile R.
clotrimazole for peak identification CRS (containing
impurities A, Band F) in I mL of acetonitrile R and dilute to
Reference solution (c) Dissolve 5.0 mg of imidazole CRS
CnH 17CIN2 344.8 23593-75-/ (impurity D) and 5.0 mg of c/otrimazole impurity E CRS in
acetonioile R and dilute to 100.0 mL with the same solvent.
Action and use Dilute 1.0 mL of the solution to 25.0 mL with acetonioile R.
Antifungal. Colmnn:
Preparations - size: /;;;;: 0.15 m, "";;;;: 4.6 mm;
Clotrimazole and Hydrocortisone Acetate Cream - stationary phase: end-capped extra-dense bonded octylsilyl silica
gelfor chromatography R (5 urn);
Clotrimazole Cream
Clotrimazole Eye Drops
Mobile phase:
Clotrimazole Pessaries - mobile phase A: dissolve 1.0 g of potassium dihydrogen
Clotrimazole Vaginal Tablets phosphate Rand 0.5 g of tetrabutylammonium hydrogen
""Ell _~ _ sulfate R/ in water for chromatography R and dilute to
1000 mL with the same solvent;
DEFINITION - mobile phase B: acetonirriJe Rl;
1-[(2-Chlorophenyl)diphenylmethyl]-IH-imidazole.
Time Moblle phase A Moblle phase B
Content
(min) (per cent JIm (per cent Vm
0-3 75 25
CHARACTERS 3 ~ 25 75 --> 20 25 --> 80
Appearance 25 - 30 20 80
White or pale yellow, crystalline powder.
Solubility Flow rate 1.0 mUmin.
Practically insoluble in water, soluble in ethanol (96 per cent) Detection Spectrophotometer at 210 run.
and in methylene chloride.
/nje<:tion I 0 ~.
IDENTIFICATION Identification of impunties Use the chromatogram supplied
Pirst identification: B. with dotrimazole for peak identification CRS and the
Second identification: A, C. chromatogram obtained with reference solution (b) to
A. Melting point (2.2./4): 141 "C to 145 "C. identify the peaks due to impurities A, Band Fi use the
B. Infrared absorption spectrophotometry (2.2.24). chromatogram obtainedwith reference solution (c) to identify
the peaks due to impurities D and E.
Comparison ckmimazo/e CRS.
Relativeretention With reference co c1otrimazole (retention
C. Thin-layer chromatography (2.2.27). time = about 12 min): impurity D = about 0.1;
Test solution Dissolve 50 mg of the substance to be impurity F;;;;: about 0.9; impurity B = about 1.1;
examined in ethanol (96 per cent) R and dilute to 5 mL with =
impurity E about 1.5; impurity A about 1.8. =
the same solvent.
Reference solution Dissolve 50 mg of cJom"mazoie CRS in - resolution: minimum 4.0 between the peaks due to
ethanol (96 per cent) R and dilute to 5 mL with the same impurity F and clotrimazole.
solvent.
Limits:
Plate TLC silica gelF254 plate R. - impurities A, B: for each impurity, not more than twice the
JWobile phase concentrated ammonia Rl, propanol R, tduene R area of the principal peak in the chromatogram obtained
(0.5: 10:90 VIVIV). with reference solution (a) (0.2 per cent);
Applicadon I0 ~. - impurities D, E: for each impurity, not more than the area
Devdopmem Over 2/3 of the plate. of the corresponding peak in the chromatogram obtained
Drying In air. - impun'ty F: not more than the area of the principal peak in
Detection Examine in ultraviolet light at 254 nrn. the chromatogram obtained with reference solution (a)
Results The principal spot in the chromatogram obtained (0.1 per cent);
with the test solutionis similar in position and size to the - unspecified impurities: for each impurity, not more than the
principal spot in the chromatogram obtained with the areaof the principal peak in the chromatogram obtained
reference solution. with reference solution (a) (0.10 per cent);
TESTS - total: not more than 5 times the area of the principal peak
Related substances
(0.5 per cent);
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1-640 Cloxacillin Sodium 2022

(0.05 per cent).
an oven at 105 "C.
Maximum 0.1 per cent, determined on 1.0 g. F. I-(triphenylmethyl)-IH-imidazole (dechloroclotrimazole).
ASSAY
___________________ Ph,,,
Dissolve 0.300 g in 80 mL of anhydrous acetic add R. Using
0.3 mL of naphtholbenze-in solution R as indicator, titrate with
O. J M perchloric acid until the colour changes from brownish-
yellow to green. Cloxacillin Sodium
I mL of 0.1 M perchloric add is equivalent to 34.48 mg (ph. Bur. monograph 0661)
of C22H17ClN2.
STORAGE
IMPURITIES
Specified impurities A, B, D, E, F.
."'0
present at a sufficient level, be detected by oneor other 0/the tests
in the mmograph. They are limited ~ thegeneral acceptance C 19H17CIN,NaO,S,H,O 475.9 7081-44-9
eriterim for otherfumpedjied impurities and/or ~ the general
monograph Substances for pharmaceutical use (2034). It is Action and use
therefore not n«essary to identify these impun"ties far Penicillin antibacterial.
demonstrau'on 0/compliance. See also 5.10. Control of impmilies
in substances for phannaceutical use) C. Ph,,, _
DEFINITION
9i6
Sodium (2S,5R,6R)-6-[[[3-(2-chlorophenyl)-5-methyl-I,2-
I"" oxawl-4-yllcarbonyl]amino]-3,3-dimethyl-7-oxo-4-thia-l-
~
azabicyclo[3.2.0]heptane-2<arboxylate monohydrate.
: OH Semi-synthetic productderived from a fermentation product.
Content
A. (2-chlorophenyl)diphenylmethanol, 95.0 per cent to 102.0 per cent (anhydrous substance).
CHARACTERS
Appearance
While or almost white, hygroscopic, crystalline powder.
Solubility
Freelysoluble in waterand in methanol, soluble in ethanol
(96 per cent).
IDENflFICATION
B. 1-[(4<hlorophenyl)diphenylmethyl]-1 H-imidazole,
Second identification: BJ C, D.
W?oCI CI
C. l-chloro-2-(chlorodiphenylmethyl)benzene,
Preparation Discs.
Comparison c/oxacillin sodium CRS.
Reference solutwn (a) Dissolve 25 mg of c/oxaciUin
sodium CRS in 5 mL of water R.
Reference solution (b) Dissolve 25 mg of cloxaciOin
D. IH-imidazole (imidazole), sodium CRS, 25 mg of di<loxaciIIin sodium CRS and 25 mg of
f/uc/oxaciOin sodium CRS in 5 mL of water R.
Plate TLC sifanised silica gelplateR.
Mobil, phase Mix 30 volumes of acetone Rand 70 volumes
of a 154 gIL solution of ammonium acetate R, then adjust to
pH 5.0 with glacial acetic add R.
E. (2-chlorophenyl)phenylmethanone Application I ~L.
(2-chlorobenzophenone), Development Over a path of 15 em.
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2022 Cloxacillin Sodium 1-641
Drying In air. Limits:

Detection Expose to iodine vapour until the spots appear; - atl)' impun·ty: not more than the area of the principal peak
examine in daylight. in the chromatogram obtained with reference solution (b)
System suitability Reference solution (b): (1.0 per cent);
Results The principal spot in the chromatogram obtained (5.0 per cent);
with the test solution is similar in position, colour and size to - disregard limit. 0.05 times the area of the principal peak in
the principal spot in the chromatogram obtained widl the chromatogram obtained with reference solution (b)
reference solution (a). (0.05 per cent).
C. Place about 2 mg In a test-tube about 150 mm long and
N,N-Dlmethylanillne (2.4.26, Method B)
15 mm in diameter. Moisten with 0.05 mL ofwattr Rand
Maximum 20 ppm.
add 2 mL of su!fi<ric acid-formaldehyde reagent R. Mix the
contents of the tube by swirling; the solution is slightly 2-Ethylhexanoic acid (2.4.28)
greenish-yellow. Place the test-tube in a water-bath for Maximum 0.8 per cent mlm.
1 min; me solution becomes yellow. Water (2.5.12)
D. It gives reaction (a) of sodium (2.3.1). 3.0 per cent to 4.5 per cent, determined on 0.300 g.
TESTS Bacterial endotoxins (2.6.14)
Solution S Less than 0.20 IU/mg, if intended for use in the manufacture
Dissolve 2.50 g in carbon dioxide-free waterR and dilute to of parenteral preparations without a further appropriate
25.0 mL with me same solvent. procedure for the removal of bacterial endotoxins,
Appearance of solution ASSAY
Solution S is clear (2.2.1) and its absorbance (2.2.25) at liquid chromatography (2.2.29) as described in the 'est for
430 run is not greater than 0.04. related substances with the following modifications.
pH (2.2.3) Injection Test solution (b) and reference solution (a).
5.0 to 7.0 for solution S. System suitability:
Specific optical rotation (2.2.7) - repeatability: maximum relative standard deviation of
+ 160 rc + 169 (anhydrous substance). 1.0 per cent after 6 injections of reference solution (a).
Dissolve 0.250 g in water R and dilute to 25.0 mL with the Calculate the percentage content of CI9H17CINJNa05S from
same solvent. the declared content of cloxadllin sodium CRS.
liquid chromatography (2.2.29). In an airtight container, at a temperature not exceeding
Test solution (a) Dissolve 50.0 mg of the substance to he 25°C. If the substance is sterile, store in a sterile, airtight,
examined in the mobile phase and dilute to 50.0 mL with tamper-evident container,
rest solutUm (b) Dilute 5.0 mL of test solution (a) to
Reference solution (a) Dissolve 50.0 mg of c/oxaciOin
the mobile phase. Dilute 5.0 mL of this solution to 50.0 mL
A. (4S) -2-[carboxy [[[3-(2-chlorophenyl)-5-methyl-1,2-oxazol-
Reference solution (c) Dissolve 5 mg ofjlucloxaciOin
4-yljcarbonyl] aminojmethyl]-5,5-dimethyl-1 ,3-
sodium CRS and 5 mg of c/oxaciOin sodium CRS in the mobile
thiazolidine-4-carboxylic acid (penicilloic acid of
cloxacillin),
Column:
- size: 1= 0.25 m, 0 = 4 mm;
- stationary phase: octade<yfsilyl silica gelfor chromatography R
(5 urn),
Mobile phase Mix 25 volumes of autonitrik Rand
phosphate R adjusted to pH 5.0 with dilute sodium hydroxide
solution R.
Flow rate 1.0 mUmin. B. (2RS,4S)-2-[[[[3-(2-chlorophenyl)-5-methyl-I,2-oxazol-4-
Detection Spectrophotometer at 225 run. yljcarbonyljamino]methyl]-5,5-dirnethyl-1,3-thiazolidine-
Injection 20 ~L of test solution (a) and reference 4-carboxylic acid (penilloic acid of cloxacillin),
Run rime 5 times the retention time of cloxacillin.
cloxacillin (I'" peak) and flucloxacillin (2nd peak).
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1-642 Clozapine 2022
Solubility
Practically insoluble in water, freely soluble in methylene
chloride, soluble in ethanol (96 per cent). It dissolvesin
dilute acetic acid.
IDENTIFICATION
C. (2S,5R,6R)-6-amino- 3,3-dimethyl-7-oxo-4-thia-l- A. Melting point (2.2.14): 182 "C to 186'C.
(6-aminopenicillanic acid),
Comparison doeapine CRS.
TESTS
Related substances
C1!.'
N_
,
o "" CO,H
So/vent mixture waterR, methanol R2 (20:80 VII').
Sduuon A Dissolve 2.04 g of potassium dihydrogen
CH, phosphate R in 1000 mL of waterR and adjust to
pH 2.4 ± 0.05 with dilute phosphoric. acidR.
D.3-(2-chlorophenyl)-5-methyl-I,2-oxazole-4-carboxylic
Test solution Dissolve75 mg of the substance to be
acid, examined in 80 mL of methanol R2 and dilute to 100 mL
with water R.
o H~.
GO,H Reference solution (a) Dilute 1.0 mL of the test solution to
Clfr
, 0
0 NH#S
~H HH
~~: 10.0 mL with the solvent mixture. Dilute 1.0 mL of this
Reference sdution (b) Dissolve the contents of a vial of
N_ N CH3
dozapine for peak identification CRS (containing
""" NH"#S
H H
CH, impurities A, B, C and D) in 1.0 mL of the solvent mixture.
CH3 0 Column:
- size: I:::;; 0.125 m, 0 :::;; 4.6 mm;
E. (2S,5R,6R)-6-[[[(2S,5R,6R)-6-[[[3-(2-chlorophenyl)-5- - stationary phase: end-capped octadecylsilYl silica gelfor
methyl-I,2-oxazol-4-yllcarbonyl] amino]-3,3-dimethyl-7- chromatography R (5 J1IIl).
oxo-4-thia-I-azabicyclo[3.2.0] hept-2-yl]carbonyl] amino]- Mobile phase:
3,3-dimethyl-7-oxo-4-thia-I-azabicyclo[3.2.0] heptane-2- - mobile phase A: acetonitrile for chromatography R,
carboxylic acid (6-APA cloxacillin amide). methanol R2, solution A (1:1:8 VIVII');
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ POE" - mobile phase B: acetonilrile for chromasography R,
methanol R2, solution A (4:4:2 VIVII');
Time Mobile phsse A Mobile phase B

(min) (per cent VIV) (per cent VIJI)
Clozapine 0-4 100 0
4 - 24 100 ..... 0 0 ..... 100
24 - 29 0 100
cr r N .... CH3
Q-oN,>
HN
N~
f'
Floro rate 1.2 mIlmin.
Deuaion Spectrophotometer at 257 nm.
Injection 20 ~L.
~
with clozapine for peak identification CRS and the
326.8 5786-21-0 identify the peaks due to impurities A, B, C and D.
Action and use Relativeretention With reference to clozapine (retention
Dopamine 1?4 receptor antagonist; neuroleptic. time = about II min): impurity C = about 0.9;
impurity D:::;; about 1.1; impurity A = about 1.6;
Preparation
Clozapine Oral Suspension
impurity B =about 1.7.
SystemsU1iabjJity Reference solution (b):
POE" ~ _ - resolution: minimum 2.5 between thepeaks due to
impurity C and dozapine;
DEFINITION
- the chromatogram obtained with reference solution (b) is
8-Chloro-ll-(4-methylpiperazin-I-yl)-5H-dibenzo[b,e]
similar to the chromatogram supplied with dosapine for
[1,4]diazepine.
peak identlfiauion CRS.
Content Limits:
99.0 per cent to 101.0 per cent (dried substance). - correction factor: for the calculation of content, multiply the
CHARACTERS peak area of impurity 0 by 2.7;
Appearance
Yellow, crystalline powder.
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2022 Cocaine 1-643
- impun"lY A: not more than the area of the principal peak

in the chromatogram obtained with referencesolution (a)
(0.1 per cent);
- impun"ties B. D: for each impurity, not more than twice
- impun"ty C: not more than 3 times me area of the
principal peak in the chromatogram obtained with D. 1-[2-[(2-amino-4-chlorophenyl)amino)benzoyl]-4-
reference solution (a) (0.3 per cent); merhylpiperazine.
- unspecified impurities: for each impurity, not more than the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I
in me chromatogram obtained with reference solution (a)
(0.6 per cent);
Cocaine
the chromatogram obtained with referencesolution (a)
(0.05 per cent).
an oven at 105°C.
ASSAY.
Dissolve 0.100 g in 50 mL of anhydrous acetic add R. Titrate
with 0.1 M perchkmc acid, determining the end-point
potentiometrically (2.2.2fJ).
I rnL of 0.1 M perchloric acid is equivalent '0
16.34 mg
303.4 5tJ..36-2
of ClsH19ClN4. Action and use
IMPURITIES Local anaesthetic.
Specified impurities A, B, C) D.
DEFINITION
Cocaine is methyl (IR,2R,3S,5S)-3-(benzoyloxy)-8-methyl-8-
azabicyclo[3.2.I]oetane-2-earboxylate and may be obtained
from the leaves of Erythroxylum coca Lam. and other species
of Erythroxylum or by synthesis. It contains not less than
98.0% and not more than 101.0% ofC 17 H 21 N Ov calculated
with reference to the dried substance.
A. 8-ehloro-5, lO-dihydro-llH-dibenzo[b,e) [1,4]diazepin-ll- CHARACTERISTICS

Colourless crystals or a white, crystalline powder. Slightly
one,
volatile.
PracricaUy insoluble in Waler, freely soluble in ethanol (96%)
and in ether, soluble in arachis oil; slightly soluble in liquid
paraffin.
IDENTIFICATION
The infrared absorption spearum, Appendix II A, is concordant
with the reference spectrum of cocaine (RS 071).
TESTS
Melting point
96° to 98°, Appendix V A.
B. II, II' -(piperazine-I,4-diyl)bis(8-chloro-5H-dibenzo[b,e] Specific optical rotation
[1,4]diazepine), In a 2.4% wlv solution in O.IM hydrochloric acid, -79 to -81,
calculated with reference to the dried substance,
Appendix V F.
Related substances
Carry out the method for liquidchromatography,
Appendix ill D, using the following solutions.
(1) 0.05% wlv of the substance being examined in the mobile
phase
C. 8-chloro-II-(piperazin-I-y1)-5H-dibenzo[b,e] [1,4] (2) Dilute I volume of solurion (I) to 50 volumes with the
diazepine, mobile phase, dilute 5.0 rnL of this solurion to 100.0 mL
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1-644 Cocaine Hydrochloride 2022
(3) Dissolve 25 mg of the substance being examined in

O.OIM sodium hydroxide and dilute to 100.0 mL with the
same solvent. Allow the solution to standfor 15 minutes.
(a) Use a stainless steel colwnn (15 em x 4.6 mm) packed
with base-deactivated octadetylsilyl silica gelfor chromatography
(5 um) (Wate", Symmetry is suitable). B. bis[(IR,2R,3S,5S)-2-(methoxycarbonyl)-8-methyl-8-
azabicyclo[3.2.I)oct-3-yl) (1.,2<,3/,4/)-2,4-
(b) Use isocratic elution and the mobilephase described
diphenylcyclobutane-I,3-dicarboxylate (c-rruxilllne),
below.
(c) Use a flow rate of 1 mL per minute.
(d) Use a colwnn temperature of 35'.
(e) Use a detection wavelength of216 run.
(I) Inject 20 ~L of each solution.
MOBILE PHASE
I volume of In'elhylamine, 200 volumes of tetrahydrofuran,
860 volumes of acetonitrile and 959 volumes of water. C. bis[(IR,2R,3S,5S)-2-(methoxycarbonyl)-8-methyl-8-
SYSTEM SUITABILITY azabicyclo[3.2.1]oet-3-yl) (I r,2<,3t,4')-3,4-
diphenylcyclobutane-I,2-dicarboxylate (~-truxilline).
The test is not valid unless, in the chromatogram obtained
with solution (3), the resolution factor between the peaks due
to cocaine (retention time, about 7 minutes) and the
degradation product is at least 5.0.
LIMITS
Cocaine Hydrochloride
In the chromatogram obtainedwith solution (1): (ph. Bur. monograph 0073)
the area of any peakeluting after the principal peakis not
greater than the area of the peak.in the chromatogram
obtained with solution (2) (0.1%);
the sum of the areas of any seamdasy peaks is not greater than • HCI
5 times the area of the principal peak in the chromatogram
obtained with solution (2) (0.5%).
Disregard any peak with an area less than 0.5 times the area
of the principal peak in the chromatogram obtained with 339.8 53-21-4
solution (2) (0.05%).
Loss on drying Action and use
When dried. to constant weight at 80°, loses not more than Local anaesthetic.
0.5% of its weight, Appendix IX D. Use I g. Preparations
Sulfated ash Adrenaline and Cocaine Intranasal Solution
Not more than 0.1 %, Appendix IX A. Cocaine Eye Drops
ASSAY Coca.ine Paste
Carry out Method I for non-aqueous titration, PhE", _
Appendix VIII A, using 0.7 g dissolved in 50 mL of
l,4-dioxan and crystal violet solution as indicator. Each mL of DEFINITION
O.IM perchloric acid VS is equivalent to 30.34 mg of Methyl (IR,2R,3S,5S)-3-(benzoyloxy)-8-methyl-8-azabicyclo
C 17H21N04 · [3.2.lloctane-2-carboxylate hydrochloride.
STORAGE Content
Cocaine should be stored protected from light. 98.5 per cent to 101.0 per cent (dried substance).
IMPURITIES CHARACTERS
Appearance
crystals.
Solubility
Very soluble .in water, freely soluble in alcohol, slightly
soluble .in methylene chloride.
mp
o About 197°C, with decomposition.
R'O~Ar IDENTIFICATION
A. methyl (JR,2R,3S,5S)-8-methyl-3-[[(E)-3- Second identification: A, C, D, E.
phenylpropenoyl]oxy]-8-azabicyclo[3.2.1]octane-2- A. Dissolve 20.0 mg in 0.01 M hydrochlonc acid and dilute to
carboxylate (cinnamoyfcocaine), 100.0 mL with the same acid. Dilute 5.0 mL of the solution
to 50.0 mL with 0.01 M hydrochloric acid. Examined between
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2022 Cocaine Hydrochloride 1-645
220 run and 350 om (2.2.25), the solution shows Limits:

2 absorption maxima, at 233 om and 273 run. The specific - atry impun·ty eluting after thepn"ncipal peak: not more than
absorbance at 233 nm is 378 to 402. the area of the principal peak in the chromatogram
B. Infrared absorption spectrophotometry (2.2.24). obtained with reference solution (a) (0.1 per cent),
Comparison Ph. Eur. reference spectrum of cocaine - ectal: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution(a)
hydrochloride.
(0.5 per cent),
C. Dissolve 0.1 ginS mL of water R and add I mL of dilute - disregard limit: 0.5 times the area of the principal peak in
ammonia R2. A white precipitate is formed. Initiate the chromatogram obtained with reference solution(a)
crystallisation by scratching the wall of the tube with a glass (0.05 per cent).
rod. The crystals, washed with water R and dried in VlUUJ),
melt (2.2.14) at 96 'C to 99 'C. Loss on drying (2.2.32)
an oven at 105°C.
E. It gives the reaction of alkaloids (2.3.1).
TESTS Maximum 0.1 per cent, determined on the residue from me
Solution S test for loss on drying.
Dissolve 0.5 g in water Rand dilute to 25 mL with the same
ASSAY
solvent.
Appearance of solution hydrochloric acid and 50 mL of alcohol R. Catty out a
Solution S is clear (2.2.1}"and colourless (2.2.2, Method II). potentiometric titration (2.2.20), using 0.1 M sodium
Acidity hydroxide. Read the volume added between the 2 points of
To 10 mL of solution S add 0.05 mL of methyl red solution R. inflexion.
Not more than 0.2 mL of 0.02 M sodium hydroxide is I mL of 0.1 M sodium hydroxide is equivalent to 33.98 mg of
required to change the colourof the indicator. C 17H22CINO•.
Specific optical rotation (2.2.7) STORAGE
-70 to -73 (dried substance). Protected from light.
IMPURITIES
same solvent.
Readily carbonisable substances
To 0.2 g add 2 mL of sulfuric acid R. After 15 min, the
solution is not moreintensely coloured thanreference
solution BY, (2.2.2, Method I).
Related substances
Exarnine by liquid chromatography (2.2.29).
Testsolution Dissolve25.0 mg of the substance to be A. methyl (IR,2R,3S,5S)-8-methyl-3-[[(E)-3-
examined in the mobilephase and dilute to 50.0 mL with phenylpropenoylj oxyj-8-azabicyclo [3.2.1 joctane-2-
the mobile phase. carboxylate (cinnamoylcocaine),
Reference sdiuion (aJ Dilute 1.0 mL of the test solution to
solution to 100.0 mL with the mobile phase. . ~~- oc:;o ~ I
o
__ H)
0 ,OCH,
: H O==' H H
examined in 0.01 M sodium hydroxide and dilute to 10.0 mL
with the same solvent. Dilute 1.0 mL of the solution to H
0
[~,c_~~ -- H ' g rH'
,.-
H"
".
08\
;
N-CH
H 3
10.0 mL with 0.01 M sodium hydro.<ide. Allow the solution to ~ 0 t
stand for 15 min. I H
#
Column:
- size: 1= 0.15 m, {2) = 4.6 rom) B. bis[(IR,2R,3S,5S)-2-(methoxycarbony1)-8-methyl-8-
- stadonory phase: end-tapped octadecyIsi/y1 silica gelfor azabicyclo[3.2.1 joct-3-yl] (I r,2<,3,,41)-2,4-
chromatography R (5 pm) with a specific surface area of diphenylcyclobutane-I,3-dicarboxylate (c-rruxilline),
335 m2/g, a pore size of 10 run and a carbon loading of
19.1 per cent,
Mobile phase triethylamine R, tetrahydrofuran R, aceumitri1e R,
lOater R (0.5:100:430:479.5 VIVIVIV).
Flow rate I mUmin.
Iniection 20 f1L.
Relative retention With reference to cocaine (retention
time = about 7.4 min): degradation product = about 0.7.
C. bls[(IR,2R,3S,5S)-2-(methoxycarbonyl)-8-methyl-8-
System suitability Reference solution (b): azabicyclo[3.2.1joct-3-yl] (I r,2c,31,4')-3,4-
- resolution: minimum of 5 between the peaks due to diphenylcyclobutane-I,2-dicarboxylate (~-rruxilline).
cocaineand to the degradation product. _----'- PhE"
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1-646 Cochineal 2022
Refractive index
Cochineal About 1.449, determined at 40 "C.
DEFINITION
IDENTIFICATION
Cochineal is the dried female insect, Dactylopius coccus Costa,
A. Melting point (see Tests).
containing eggs and larvae.
B. Composition of fatty acids (see Tests).
CHARACTERISTICS
TESTS
Odour, characteristic.
Melting point (2.2. 14)
Macroscqpical Purplish black or purplish grey; about 3.5 to
23 °C to 26 °C.
5.5 mm long and 3 to 4.5 mm wide, plano-convex and
somewhat oval in outline; the convex dorsal surface is Acid value (2.5. I)
transversely wrinkled and shows about 11 segments; the flat Maximum 0.5, determined on 20.0 g.
or slightly concave ventral surface carries upon the anterior Peroxide value (2.5.5, Method A)
part two seven-jointed straight antennae, three pairs of short Maximum 5.0.
legs, each terminating in a single claw, and a mouth from Unsaponlfiable matter (2.5.7)
which projects the remains of a long filifonn proboscis; these Maximum 1.0 per cent, determined on 5.0 g.
appendages are frequently more Or less broken. Easily
A1)<alIne Impurities (2.4. I 'J)
reduced to powder, which is dark red or puce.
Microscopical Scattered irregularly over the whole dermis are
numerous solitary and grouped, short, tubular wax glands; Composition of fatty acids (2.4.22, MethodB)
within each insect are found numerous larvae, which are Refined coconut oil is melted under gentle heating to a
characterised by their proboscides appearing as two circular homogeneous liquid prior to sampling.
coils. Reference solution Dissolve 15.0 mg of tn'caprm'n CRS,
80.0 mg of'ristearin CRS, 0.150 g of tncaprin CRS, 0.200 g
TESTS
of 'ricaprylin CRS, 0.450 g of trimyris,in CRS and 1.25 g of
Colour value
m·lam;n CRS in a mixture of 2 volumes of methylene
To 0.5 g in moderately fine puroder add 60 mL of phosphate
chloride Rand 8 volumes of heptane R, then dilute to 50 mL
bufferpH 8.0 and heat on a water bath for 30 minutes. Cool,
with the same mixture of solvents heating at 45-50 °C.
add sufficient phosphate buffer pH 8.0 to produce 100 mL and
Transfer 2 mL of this mixture to a 10 mL centrifuge rube
filter. Dilute 5 mL of the filtrate to 100 mL with phosphate
with a screw cap and evaporate the solvent in a current of
briffer pH 8.0. The absorbance of the resulting solution at the nitrogen R. Dissolve with I mL of heptane R and I mL of
maximum at 530 run is not less than 0.25, Appendix II B. dimethyl carbonate R and mix vigorously under gentle heating
Foreign matter (50-60 'c). Add, while still wann, I mL of a 12 gIL solution
Complies with the test for foreign matter, Appendix XI D. of sodium R in anhydrous methanol R, prepared with the
Water-insoluble matter necessary precautions} and mix vigorously for about 5 min.
When the insects are placed in water, no insoluble powder Add 3 mL of distilled water R and mix vigorously for about
separates. 30 s. Centrifugefor 15 min at 1500 g. Inject I ~L of the
organic phase.
Ash
Not more than 7.0%, Appendix XI J. Calculate the percentage content of each fatty acid using the
I g is free from Escherichia cob; 109 is free from Salmonella,
Appendix XVI B I. ::t x
LJ x,s,C
100 per cent m/m
A x ,.I, c is the corrected peak area of each fatty acid in the test
solution:
Coconut Oil
(RefinedCoconut Oil, Ph. Eur. monograph 1410)
8001-31-8 R is the relative correction factor:
Action and use mx,r xA1,r

Excipient. Ax,rxml,r
PhElI _
for the peaks due to caproic} caprylic, capric, lauric and
DEFINITION myristic acid methyl esters.
Fatty oil obtained from the dried, solid part of the
endosperm of Cocos nudfera L., then refined. m..,._ mass oftricaproin, tricaprylin, tricaprin, trilaurin or trirnyrisrin
in the reference solution, in milligrams;
CHARACTERS ml.... mass of tristearin in the reference solution. in milligrams:
Appearance Az.>' area of the peaks due to caproic. caprylic,capric, lauric- and
myristic acid methyl esters in the reference solution;
White or almost white, unctuous mass. AI.... area of the peak due [0 stearic acid merhgl ester in the
Solubility reference solution;
Practically insoluble in water, freely soluble in methylene Ax,> = area of the peaks due to any specifiedor unspecified fatty acid
methyl esters;
chloride and in light petroleum (bp: 65-70 "C), very slightly ~ 1 for the peaks due to each of the remainingspecifiedflltty acid
soluble in ethanol (96 per cent). methyl esters or any unspecifiedfatty acid methyl ester.
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2022 Codeine 1-647
Composition of thefauy-acid fraction of the oil: Saponification value (2.5.6)

- caproic acid (RRt 0.11): maximum 1.5 per cent, 160 to 173.
- caprylic acid (RR, 0.21): 5.0 per cent to 11.0 per cent, Composition of fatty acids and fatty alcohols (2.4.22,
- capric acid (RRt 0.56): 4.0 per cent to 9.0 per cent, MethodC)
- lauric acid (RR, 0.75): 40.0 per cent to 50.0 per cent, Use the chromatogram obtained with the following reference
- myristic acid (RRI 0.85): 15.0 per cent to 20.0 per cent, solution for identification of the peaks due to the fatty
- palmiticacid (RR' 0.93): 7.0 per cent to 12.0 per cent, alcohols.
- stearic acid (RRt 1.00): 1.5 per cent to 5.0 per cent,
Reference solution Dissolve the amounts of the substances
- oleic add (RR, J.OJ): 4.0 per cent to 10.0 per cent,
listed in the following table in 10 mL of heptane R.
- linoleic acid (RR, 1.03): 1.0 per cent to 3.0 per cent,
- linolenk add (RR! 1.06): maximum 0.2 per cent,
Substance Amount (mg)
- arachidic acid (RR, 1.10): maximum 0.2 per cent)
Mtl1r~ caproafe R 10
- eicosenoic acid (RRI 1.11): maximum 0.2 per cent.
Methyl caprylate R 90
Water (2.5.32) Muh!l~ deconoue R 50
Maximum 0.1 per cent, derermlned on 1.00 g. MelhyllaurareR 20
STORAGE Melhyl IIJ,Yri5tate R 10
In a well-filled container, protected from light. Methylpalmi/ate R 10
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE.. Muhyl stearate R 10
DeamolR 10
uJUryl alcoholR 100
M)-rislyl alcohol R '0
Cet;yl alwholCRS 30
Cocoyl Caprylocaprate Sfe(1)'lokoho/ CRS 20
(ph. Bur. monograph J4JJ)

Consider the sum of the areas of the peaks due to the fatty
Action and use adds listed below to be equal to 100 and the sum of the
Excipient. areas of the peaks due to the fatty alcohols listed below to be
PhE.. _ equal to 100.
Composition of thefa~ acidfraction of the substance:
DEFINlTION - caproic acid: maximum 2.0 per cent,
.Mixture of esters of saturated C 12 - CIS alcohols with - caprylic acid: 50.0 per cent to 80.0 per cent)
caprylic (octanoic) and capric (decanoic) acids obtained by - capn·c acid: 20.0 per cent 10 50.0 per cent,
the reaction of these acids with vegetable saturated fatty - lauric acid: maximwn 3.0 per cent,
alcohols. - myristic acid: maximum 2.0 per cent.
CHARACTERS Composition of thefOllY akoholfraaion of the substance:
Appearance - capric a/rohol: maximum 3.0 per cent,
Slightly yellowish liquid. - laurylalcohol: 48.0 per cent to 63.0 per cent,
Solubility - myrislyl akohol: J8.0 per cent 10 27.0 per cent,
Practically insoluble in water, miscible with ethanol - cetyl a/rohol: 6.0 per cent to 13.0 per cent)
(96 per cent) and with liquid paraffin. - stearyl akohol: 9.0 per cent to 16.0 per cent.
Relative density Water (2.5.12)
About 0.86. Maximum 0.1 per cent) determined on 5.00 g.
Refractive index Total ash (2.4.J6)
About 1.445. Maximum 0.1 per cent, determined on 1.0 g.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE..
Viscosity
About II ml'a-s.
IDENTIFICATION
A. Freezing point (2.2.J8): maximum 15 'C.
Codeine Monohydrate
Comparison cocoyl caprylocaprate CRS. (Ph. Bur. monograph 0076)
C. Composition of fatty acids and fatty alcohols (see Tests).
TESTS
Appearance
The substance to be examined is not more intensely coloured
than reference solution Y j (2.2.2" klethod l).
Acid value (2.5. J)
M.aximum 0.5, determined on 5.00 g.
Maximum 5.0, determined on 4.0 g. Add 5.0 mL of 317.4 6059-47-8
acetylating reagent.
Action and use
Iodine value (2.5.4, MethodA) Opioid receptor agonist; analgesic.
Maximnrn 1.0.
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1-648 Codeine 2022
PhE" _ Time Mobile phase A Mobile phase B

(min) (per cent V/V) (per cent VIJI)
DEFINITION 0-5 o
'00
4,5a-Epoxy-3-methoxy-17-methyl-7,8-<!idehydromorphinan- 5 -7.33 100 --> 93 0~7
6ct-ol monohydrate. 7.33 - 10.33 93 --> 67 7 -0 33
Content 10.33 - 12 67 33
CHARACfERS Flow rale 1.0 mUmin.
Appearance Detection Spectrophotometer at 280 ron.
Whiteor almost white, crystalline powder or colourless Injection 3 ~L.
crystals. Identification of impuniies Use the chromatogram supplied
Solubility with codeine for system suitab11ity CRS and the chromatogram
Slightly soluble in water, freely soluble in ethanol obtainedwith reference solution (b) to identify the peaks due
(96 per cent). to impurities A, B, C, D, E, F, G, H and I.
IDENTIFICATION Relative retention Wirh reference to codeine (retention
First ickmificatitm: A, B, E. time :: about 6 min): impurity B == about 0.3;
Second identification: A, G, D, E. impurity E == about 0.4; impurity F == about 0.8;
impurity H = about 0.9; impurity C == about 1.2;
A. Melting point (2.2.14): 155. °C to 159 °C.
=
impurity I about 1.4; impurity D about 1.45; =
B. Infrared absorption spectrophotometry (2.2.24). =
impurity A about 1.5; impurity G about 1.6. =
Comparison codeine CRS. System suitability Reference solution (b):
C. To about 10 mg add I mL of sulfuric acid Rand 0.05 mL - resolution: minimum 2.5 between the peaksdue to
offerri< chloride solutian R2 and heat on a water-bath. A blue impurities F and H; minimum 1.5 between the peaks due
colour develops. Add 0.05 mL of nitric acid R. The colour to impurities D and A.
changes to red. Calculation of percentage contents:
D. It gives the reaction of alkaloids (2.3.1). - correction factors: multiply the peak areas of the following
E. Loss on drying (see Tests). impurities by the corresponding correction factor:
impurity C = 0.7; impurity G = 0.2; impurity I = 1.3;
TESTS
- for each impurity, use the concentration of codeine
Appearance of solution monohydrate in reference solution (a).
The solution is clear (2.2.i) and colourless (2.2.2,
Limits:
Method II).
Dissolve 50 mg in water R and dilute to 10.0 mL with the - impurity H: maximum 0.25 per cent;
same solvent. - impurities C, D, E: for each impurity, maximum
Specific optical rotation (2.2.7) 0.2 per cent;
-146 to -142 (dried substance). - impun·ties B, F, 0, I: for each impurity, maximum
Dissolve 0.50 g in ethanol (96 percen!! R and dilute to 0.15 per cent;
25.0 mL with the same solvent. - unspecified impurities: for each impurity, maximum
0.10 per cent;
Related substances
Solution A 0.5 per cent VIV solution of phosphoric acidR.
Loss on drying (2.Z.32)
4.0 per cent to 6.0 per cenr, determined on 1.000 g by
examined in solutionA and dilute to 50.0 mL with
drying in an oven at 105°C.
solution A.
Sulfated ash (Z.4.14)
Reference solutian (a) Dilute 2.0 mL of the test solution to
100.0 mL with solution A. Dilute 1.0 mL oftltis solution to
10.0 mL with solution A. ASSAY
Reference solutian (b) Dissolve 3 mg of codeine for system Dissolve 0.250 g in 50 mL of anhydrous acetic acidR. Titrate
suitabl1iiy CRS (containing impurities A, B, C, D J E, F, G, H with 0.1 M perchiotic acid, determining the end-point
and I) in 1 mL of solution A. potentiometrically (2.2.20).
Column: I mL of 0.1 M perchlonc acid is equivalent to 29.94 rng
- size: 1= 0.075 m, 0 = 3.0 mm; of C,sH21N03'
- statianary phase: end-capped octadecy/silyl mul,;-Iayered STORAGE
organo,ilica polymer for chroma/Qgraphy R (1.9 urn). Protected from light.
IMPURITIES
Mobile phase:
- mobile phase A: mix 4 volumes of acetonitrile Rand Specified impurities A, B, C, D, E, F, GJ H, 1.
96 volumes of a 20 gIL solution of glacial acetic acidR Other deuaabie impurities (the following sub,tances would, if
previously adjusted to pH 4.5 with a 500 gIL solution of present at a sufficient level, be detected by oneor other oj the tests
sodium hydroxide R; in the monograph. They are limited by thegeneral acceptance
- mobile phase B: acetonitrile R; criterion for other/unspecified impurities and/or by thegeneral
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2022 Codeine 1-649

in substances for pharmaceutical use) JJ K, L, 1'1'1
F. 4,5a-epoxy-3-methoxy-l7-methyl-7,8-
didehydromorphinan-6a,14-dio~
A. 4,5a-epoxy-3,6a-dimethoxy-17-methyl-7,8-
didehydromorphinan (methylcodeine),
,
CH,
H N
@
HO o: H H OH
G. 4,5a-epoxy-3,6-dimethoxy-17-methyl-6,7,8,14-
tetradehydromorphinan (thebaine),
B. 4,5a-epoxy-17-methyl-7,8-didehydromorphinan-3,6a-diol
(morphine),
H. 4,5a-epoxy-3-methoxy-7,8-didehydromorphinan-6a-ol
(norcodelne),
N H
I
CH,
C. 4,5C(:4',5'a.-diepoxy-3,3'-dimelhoxy-17,17' -dirnethyl-
7,7',8,8 1-tetradehydro-2,2'-bimorphinan-eo.ti'a-diol
(codeine dimer),
,
CH,
H N 1. 4,5a-epoxy-3-methoxy-17-methyl-7,8-
_o~
didehydromorphinan-6-one (codeinone),
HOHH. O
~ 0, O"'H H
O
'
OH
~lV CH
3
N H
I
CH,
D. 4,5a-epoxy-2-[(4,5a-epoxy-6a-hydroxy-17-methyl-7,8- J. (J 7RS)-4,5a-epoxy-6a-hydroxy-3-methoxy-17-methyl-7,8-
didehydromorphinan-3-yl)oxy]-3-methoxy-17-methyl-7,8- didehydromorphinan-17 -oxide (codeine N-oxide),
didehydromorphinan-6a-ol (3-0-(codein-2-yl)morphine),
K. 4,5a-epoxy-14-hydroxy-3-methoxy-17-methyl-7,8-
E. 4,5a-epoxy-3-methoxy-17-methyl-7,8- didehydromorphinan-6-one (J4-hydroxycodeinone),
didehydromorphinan-fic, JOS-diol,
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1-650 Codeine Hydrochloride 2022
water R and initiate crystallisation, if necessary, by scratching

the wall of the tube with a glass rod and cooling in iced
water. Wash the precipitate with water R and dry at
100-105 "C. It melts (2.2.15) at 155"C to 159 "C.
C. To about 10 mg add I mL of su/furic acid Rand 0.05 mL
ofJenU chloride sdution R2 and heat on a water-bath. A blue
colourdevelops. Add 0.05 mL of niuic acid R. The colour
L. 4,5~-epoxy-6-methoxy-17-methyl-6, 7,8,14- changes to red.
tetradehydromorphinan-S-ol (oripavine), D. Solution S gives reaction (a) of chlorides (2.3.1).
E. It gives the reacrion of alkaloids (2.3.1).
,
CH,
F. Water (see Tests).
ri\l", H N
HO H...o _ OCH, TESTS
~o~
H3CO '=lV O....H OH
Solution S
Dissolve 2.00 g in carbon dioxide-free water R, prepared from
dun·Oed waterR, and dilute to 50.0 mL with the same solvent.
,
N H Appearance of solution
SolutionS is clear (2.2.1) and not moreintensely coloured
CH,
than reference solution Y6 (2.2.2J Method II).
M. 7,7'-oxybis(4,5~-epoxy-3-methoxy-17-methyl-6,7,8,14- Acidity or alkalinity
tetradebydromorphinan-ri-ol) (7,7'-oxybis(6-0- To.5 mL of solution S add 5 mL of carbon dioxide-free
demethylrhebainej), water R. Add 0.05 mL of methyl red solu,;on Rand 0.2 mL of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<r 0.02 M hydrochkJri< acid; the solution is red. Add 0.4 mL of
0.02 M sodium hydroxidej the solution becomes yellow.
Codeine Hydrochloride Dilute 5.0 mL of solution S to 10.0 mL with water R.
Related substances
(Codeine Hydrochloride Dihydrate, Ph. Eur.
monograph 1412)
So/utWn A 0.5 per cent VIV solution of phosphori< acidR.
,
CH, Test solution Dissolve 0.190 g of the substance to be
Hco~t
examined in solution A and dilute to 50.0 mL with
solution A.
Hel . 2~O
Reference so/utWn (a) Dilute 2.0 mL of the test solution to
a H H 10.0 mL with solution A.
Reference solution (b) Dissolve 3 mg of codeine for system
C18HnClNO,,2H20 371.9 5916-29-0 suitabiJii)' CRS (containing impurities A, B, C, D, E, FJ G, H
and I) in I mL of solution A.
Opioid receptor agonist; analgesic. - size: I;;;; 0.075 m, 0 = 3.0 mm;
PIlE" _ - statWnary phase: end-capped octade<y/silyl mul,;-Iayered
organosilica polymer for chromaUJgraphy R (1.9 pm).
DEFINITION - tempemture: 40 °C.
4,5~-Epoxy-3-methoxy-17-methyl-7,8-didehydromorphinan
Mob"ephase:
6~-01 hydrochloride dihydrate. - mobile phase A: mix 4 volumes of autonjtn·le Rand
Content 96 volumes of a 20 gIL solution of glacial acetic acidR
99.0 per cent to 101.0 per cent (anhydrous substance). previously adjusted to pH 4.5 with a 500 gIL solution of
CHARACTERS
sodium hydroxide R;
Appearance
White or almost white, crystalline powder or smaU, colourless
Time Moblle phase A Mobile phase B
crystals. (min) (per cent VIV) (per cent VIJI)
Solubility 0-5 \00 0
Soluble in water, slightly soluble in ethanol (96 per cent), s -7.33 100 --> 93 o~ 7
practically insoluble in cyclohexane. 7.33·10.33 93 --> 67 7 --> 33
IDENTIFICATION 10.33 - 12 67 3J
First identification: A" D, F.
Second identification: B, C, D, E, F. Flow rate 1.0 mUmin.
A. Infrared absorption spectrophotometry (2.2.24). Detection Spectrophotometer at 280 nm.
Comparison codeine hydfYXhkJride dihydrate CRS. Injection 3 ~L.
B. To 5 mL of solution S (see Tests) add I mL of a mixture Identification oj impurities Use the chromatogram supplied
of equal volwnes of strong sodium hydroxide solution Rand with codeine for system suiulbiJity CRS and the chromatogram
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2022 Codeine Hydrochloride 1-651
CH,
obtained with referencesolution (b) to identify the peaks due I
to impurities A, B, CJ D, E, F, G, Hand 1. H N
Q3lL
Relative retention With reference to codeine (retention
impurity E = about 0.4; impurity F = about 0.8;
impurity H = about 0.9; impurity C = about 1.2; HCO
a H H
impurity I = about 1.4; impurity D = about 1.45;
impurity A = about 1.5; impurity G = about 1.6. A. 4,51X-epoxy-3,61X-dimethoxy-17-methyl-7,8-
System suitability Reference solution (b): didehydromorphinan (methylcodeine),
impurities F and H; minimum 1.5 between the peaks due
to impurities D and A.
Calculation of percenUlge contents:
impurity C = 0.7; impurity G = 0.2; impurity I = 1.3;
hydrochloride dihydrate in reference solution (a).
Limits: B. 4,51X-epoxy-17-methyl-7,8-didehydromorphinan-3,61X-diol
- impurity A: maximum 1.0 per cent; (morphine),
- impun"ty H: maximwn 0.25 per cent;
- impurities C~ D~ E: for each impurity, maximum ,
CH,
N
0.2 per cent;
- impuruies B~ F~ G, I: for each impurity, maximum HO...
0.15 per cent;
0.10 per cent;
\
,
N H
CH,
Sulfates (2.4.13)
Maximum 0.1 per cent. C. 4,51X:4',5'lX-diepoxy-3,3'-dimethoxy-17,17' -dimethyl-
Dilute 5 mL of solution S to 20 mL with disriUed warer R. 7)7' ~8~8'-tetradehydro-2,2' -bimorphinan-oo,6'ee-dlol
Water (2.5.12) (codeine dimer),
ASSAY
,
CH,
Dissolve 0.300 g in a mixrure of 5 mL of 0.01 M hydrochloric

acid and 50 mL of ethanol (96 per cen!i R. Titrate with 0.1 M
H H 0 o~H\N
potentiometrically (2.2.20). Read the volume added between HO.~
...... - _ .., ~"OH
the 2 points of inflexion. \. j 0, o' H H

H CH3
1 mL of 0.1 M sodium hydroxide is equivalent to 33.59 mg
of C,.H"CINO,. \
STORAGE
,
N H
CH,
IMPURITIES D. 4,51X-epoxy-2-[(4,51X-epoxy-6IX-hydroxy-17-methyl-7,8-
Specified impurities A, B~ C~ D~ E~ F~ G~ H~ 1. didehydromorphinan-3-yl)oxy]-3-methoxy-17-methyl-7,8-
Other detectable impurities (thefollowing substances would~ If didehydromorphlnan-tic-ol (3-D-(codein-2-yl)morphine),
present at a sujficiem leve/~ be detected by Qtle or other of the tests
in themonograph. They are limited by the general acceptance yH3
HO H N
criterion for other/unspecified impun'M and/or by thegeneral
J~o"
monograph Sub,umces for pharmaceutical use (2034). It is
demonstration of compliaru:e. See also 5.10. Control of impurities
in substances for pharmaceutical use) J~ 1(, L~ M.
E. 4,51X-epoxy-3-methoxy-17-methyl-7,8-
didehydromorphinan-6IX,I01;-diol,
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1-652 Codeine Phosphate 2022
,
CH,
H N
~~
F. 4,5~-epoxy-3-methoxy-17-methyl-7,8 L. 4,5a-epoxy-6-methoxy-17-methyl-6,7,8,14-
didehydromorphinan-oc.Id-dlol, tetradehydromorphinan-3-ol (oripavine),
G. 4,S~-epoxy-3,6-dimethoxy-17-methyl-6,7,8,14
M. 7,7'-oxybis( 4,5~-epoxy-3-methoxy-17 -methyl-6,7,8,14-
tetradehydromorphinan-ri-ol) (7,7'-oxybis(6-0-
demethylrhebainej).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Codeine Phosphate ***

H. 4,S~-epoxy- 3-methoxy-7,8-didehydromorphinan-00-ol *** *•*
(norcodeine), (Codeine Phosphate Heminydmte, Ph. Eur. monograph *.*
0074)
CH3
I
N
I. 4,S~-epoxy-3-methoxy-17-methyl-7,8
406.4 41444-62-6
Action and use

Opioid receptor agonist; analgesic.
Preparations
Co-codamol Capsules
Co-codamol Tablets
J. (17 RS)-4,5~-epoxy-00-hydroxy-3-methoxy-17 -methyl-7 ,8-
Co-codamol Effervescent Tablets
didehydromorphinan 17-oxide (codeine N-oxide), Co-codaprin Tablets
Co-codaprin Dispersible Tablets
Codeine Phosphate Injection
Codeine Phosphate Oral Solution
Codeine Phosphate Tablets
Paracetamol, Codeine Phosphate and Caffeine Capsules
Paraceramol, Codeine Phosphate and Caffeine Tablets
PIlE" _
K. 4,5~-epoxy-14-hydroxy- 3-methoxy-17-methyl-7,8- DEFINITION

didehydromorphinan-6-one (14-hydroxycodeinone), 4,S,,-Epoxy-3-methoxy-17-methyl-7 ,8-didehydromorphinan-
00-01 phosphate hemihydrate.
Content
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2022 Codeine Phosphate 1-653
CHARACTERS previously adjusted to pH 4.5 with a 500 gIL solution of

Appearance sodium hydroxide R;
White or almost white, crystalline powder or small, colourless - mobile phase B: acetonitrile R;
crystals.
Solubility Thn. MobUe phase A Mobile phase B
Freely soluble in water, slightly soluble or very slightly
0-5 100 0
5 - 7.33 100 ---> 93 O~7
IDENTIFICATION 7.33 • 10.33 93 ..... 67 7 ---> 33

Fine identification: A, D, E. 10.33 - 12 67 33
Second identification: B, C, D, E, F.
A. Infrared absorption spectrophotometry (2.2.24). Flaw rate 1.0 mUmin.
Preparation Dissolve 6.20 g in 4 mL of warer R. Add I mL Detection Spectrophotometer at 280 nm.
of a mixture of equal volumes of strong sodium hydroxide l,y«tion 3 ~L.
solution R and waterR and initiate crystallisation, if necessary, Identification of impurities Use the chromatogram supplied
by scratching the wall of the tube with a glass rod and with codeine for system suitability CRS and the chromatogram
cooling in iced water. Wash the precipitate with water Rand obtained with reference solution (b) to identify the peaks due
dry at 100-105 'C. to impurities A, B, C, D, E, F, G, H and I.
Comparaison codeine CRS. Relative retention With reference to codeine (retention
B. Dissolve 0.20 g in 4 mL of walei' R. Add I mL of a time = about 6 min): impurity B = about 0.3;
mixture of equal volumes of strong sodium hj'<iroxide solution R =
impurity E about 0.4; impurity F about 0.8; =
and water R and initiate crystallisation, if necessary, by impurity H = about 0.9; impurity C = about 1.2;
scratching the wall of the tube with a glass rod and cooling in impurity I = about 1.4; impurity D = about 1.45;
iced water. The precipitate, washed with water R and dried at =
impurity A about 1.5; impurity G about 1.6. =
100-105 'C, melts (2.2.14) at 155 'C to 159 'C. System suitability Reference solution (b):
C. To about 10 mg add I mL of sulfuri< acid Rand 0.05 mL - resolution: minimum 2.5 between the peaks due to
of ferric chloride solution R2 and heat on a water-bath. A blue impurities F and H; minimum 15 between the peaks due
colour develops. Add 0.05 mL of nitric acidR. The colour to impurities D and A.
changes to red. Calculation of percentage contents:
D. Loss on drying (see Tests). - correction factors: multiply the peak areas of the following
E. Solution S gives reaction (a) of phosphates (2.3.1). impurities by the corresponding correction factor:
F. It gives the reaction of alkaloids (2.3.1). = =
impurity C 0.7; impurity G 0.2; impurity I 1.3; =
TESTS phosphate hemihyd.rate in reference solution (a).
Solution S
Limits:
Dissolve 1.00 g in carbon dioxide-free waterR prepared from
- impun'ty A: maximum 1.0 per cent;
distilled water R and dilute to 25.0 mL with the same solvent. - impurity H: maximum 0.25 per cent;
pH (2.2.3) - impurities C, D, E: for each impurity, maximum
4.0 to 5.0 for solution S. 0.2 per cent;
Specific optical rotation (2.2.7) - impun·ties B, F, G, I: for each impurity, maximum
-102 to -98 (dried substance). 0.15 per cent;
Dilute 5.0 mL of solution S to 10.0 mL with walei' R.
- unspecified ;mpun·,ies: for each impurity, maximum
0.10 per cent;
Related substances - totol: maximum 15 per cent;
Liquid chromatography (2.2.29). - reporting threshold: 0.05 per cent.
SoIurwn A 0.5 per cent VIV solution of phosphoric acid R. Sulfates (2.4.13)
Test solution Dissolve 0.190 g of the substance to be Maximum 0.1 per cent.
Dilute 5 mL of solution S to 20 mL with distiUed warer R.
solution A.
Loss an drying (2.2.32)
drying in an oven at 105°C.
10.0.mL with solution A.
Reference solution (b) Dissolve 3 mg of codeine for system ASSAY
suitability CRS (containing impurities A, B, C, D, E, F, G, H Dissolve 0.350 g in 50 mL of anhydrous acetic acid R. Titrate
and I) in 1 mL of solution A. with 0.1 M perchloric add, determining the end-point
Column:
- size: / = 0.075 m, {2) = 3.0 mm; 1 mL of 0.1 M perchlotic acid is equivalent to 39.74 mg
- stationary phase: end-capped octade<y/silyl multi-layered of C,sH"N07P.
organosilica polymer for chromatography R (1.9 pm). STORAGE
- temperature: 40 "C. Protected from light.
Mob_ephase: IMPURITIES
- mobile phase A: mix 4 volumes of acetonitrile Rand
Specified impurities A, B, C, D, E, F, G, H, 1.
96 volumes of a 20 gIL solution of glacial acetic add R
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1-654 Codeine Phosphate 2022
Otherdetectable impuriues (thefollowing substances would, if

in the monograph. Theyare limited by the general acceptanu
there/ore not necessary to identify these impurities for
in substanus forphannaceutical use) J, K, L, M.
E. 4,5~-epoxy- 3-methoxy-17-methyl-7,8-
didehydromorphinan-oc.Ino-diol,
,
CH,
H N
A.
-~-
4,5~-epoxy- 3,oo-dimethoxy-17-methyl-7,8-
didehydromorphlnan (methylcodeine),
F. 4,5~-epoxy- 3-methoxy-17-methyl-7,8-
didehydromorphinan-Sc, 14-diol,
,
CH,
H N
B.
"~OO
4,5~-epoxy-17 -methyl-7,8-didehydromOlphinan-3,oo-diol
(morphine),
G. 4,5~-epoxy- 3,6-dimethoxy-17-methyl-6,7,8,14-
H. 4,5~-epoxy-3-methoxy-7,8-didehydromorphinan-oo-ol
C. 4,5«:4',5'ct-diepoxy-3,3 /-dimedtoxy-17,17'-dimethyl- (norcodeine),
7,7',8,8'-tetradehydro-2,2'-bimorphinan-6a,6'et-diol
(codeine dimer), ,
CH,
~co
~ 0 H 0
1. 4,5~-epoxy- 3-methoxy-17-methyl-7,8-
D. 4,5~-epoxy-2-[(4,5~-epoxy-oo-hydroxy-17-methyl-7,8
didehydromorphinan-3-yl)oxy]-3-methoxy-17-methyl-7,8-
didehydromorphinan-tic-ol (3-0-(codein-2-yl)morphine),
J. (17 RS)-4,5~-epoxy-oo-hydroxy-3-methoxy-17-methyl-7,8-
didehydromorphinan-17-oxide (codeine N-oxide),
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2022 Codeine Phosphate Sesquihydrate 1-655
CHARACTERS
Appearance
White or almost white, crystalline powder or small,colourless
crystals.
Solubility
Freelysoluble in water, slightly soluble in ethanol
(96 per cent).
K. 4,5~-epoxy-14-hydroxy- 3-methoxy-17-methyl-7,8- IDENTIFICATION
didehydromorphinan-6-one (14-hydroxycodeinone), First identification: A, D, E.
Second identification: B, C, D, E.. F.
Preparation Dissolve 0.20 g in 4 mL of water R. Add I mL
of a mixture of equal volumes of strong sodium hydroxide
solution R and water R and initiate crystallisation, if necessary,
by scratching the wall of the tube with a glass rod and
cooling in iced water. Wash the precipitate with water Rand
dry at 100-105 "C. Examine the dried precipitate prepared as
L. 4,5lX-epoxy-6-methoxy-17-methyl-6,7,8,14- discs using potassium bromide R.
tetradehydromorphinan-3-o1 (oripavine), Comparison codeine CRS dried at 105 "C.
B. Dissolve 0.20 g in 4 mL of waterR. Add I mL of a
,
CH,
mixture of equal volumes of strong sodium hydroxide sdution R
H N and water R and initiate crystallisation, if necessary, by
~1~ HO H....O OCH, scratching the wall of the tube with a glass rod and cooling in
~~
""lV
H,CO - O/H OH 0
iced water. The precipitate, washed with water R and dried at
100-105 "C, melts (2.2.14) at 155"C to 159 "C.
C. To about 10 mg add I mL of sulfuric acidRand 0.05 mL
N H offeme chloride sdution R2 and heat on a water-bath. A blue
I
CH, colour develops. Add 0.05 mL of nitric acid R. The colour
changes to red.
M. 7,7'-oxybis(4,5~-epoxy-3-methoxy-17-methyl-6,7,8,14- D. Loss on drying (see Tests).
terradehydromorphinan-ti-ol) (7,7'-oxybis(6-D- E. Solution S gives reaction (a) of phosphates (2.3.1).
demethylthebaine)). F. It gives the reaction of alkalnids (2.3.1).
____________________ "'E"
TESTS
Soludon S
Codeine Phosphate Sesquihydrate pH (2.2.3)
(Ph. Bur. monograph 0075) 4.0 to 5.0 for solution S.
Specific optical rotadon (2.2.7)
Dilute 5.0 mL of solution S to 10.0 mL with water R.
Related substances
Sokuion A 0.5 pet cent VIV solution of phosphoric acidR.
Test solutio!) Dissolve 0.190 g of the substance to be
424.4 59/3-76-8 solution A.
Reference sdution (a) Dilute 2.0 mL of the test solution to
Action and use 100.0 mL with solution A. Dilute 1.0 mL of this solution to
Opioid receptor agonist; analgesic. 10.0 mL with solution A.
Preparadons Reference solution (b) Dissolve 3 mg of codeine for system
Codeine Phosphate Oral Solution suitabilily CRS (containing impurities AJ B, C, D, EJ F, G, H
Codeine Phosphate Tablets and I) in I mL of solution A.
PhE" _
Column:
- size: 1= 0.075 m, 0 = 3.0 rnrn;
DEFINITION - stationary phase: end-capped octadecyIsi/y1 multi-layered
4,5~-Epoxy-3-methoxy-17-methyl-7,8-<lidehydromorphinan- organon/ira polymer for chroma,ography R (1.9 pm).
6~-01 phosphate sesquihydrate. - temperamre: 40 °C.
Content Mobile phase:
99.0 per cent to 101.0 per cent (dried substance). - mobile phaseA: mix 4 volumesof acetonitrile Rand
96 volumes of a 20 gIL solution of glacial aulic acidR
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1-656 Codeine Phosphate Sesquihydrate 2022
previously adjusted to pH 4.5 with a 500 gIL solution of Otherdetectable impurities (thefollowing substances uoutd, if
sodium hydroxide R; present at a sufficient level, be detected by one or other of the tests
- mobile phase B: acetonitrile Rj in the monograph. They are limited by thegeneral acceptance
criterion for othertunspecified impurities and/or by thegeneral
Time Mobile phase A MobUe phase B monograph Substances for pharmaceutical use (2034). 1, is
(min) (per cent VIP) (per cent JIll? therefore not neussary to identify these impurities for
0-5 100 demonstration o/compliance. See also 5.10. Control of impurities
5 -7.33 100 -+ 93 in substances for pharmaceutical use) J, K, L, M.
7.33-10.33 93 -+ 67
10.33 - 12 67
FWw rate 1.0 mUmin.

lnjeaion 3 ~L.
ldemification of impurities Use the chromatogram supplied
with codeine for system suitability CRS and the chromatogram
A. 4,5cr-epoxy-3,6<x-dimethoxy-17-methyl-7,S-
obtainedwith reference solution (b) to identify the peaks due
to impurities A, B, C, D, E, F, G, H and I. didehydromorphinan (methylcodeine),
Relative retention With reference to codeine (retention ,
CH,
~t
time == about 6.0 min): impurity B == about 0.3; _
impurity E == about 0.4; impurity F ;;;; about 0.8;
impurity H == about 0.9; impurity C == about 1.2;
impurity I = about 1.4; impurity D == about 1.45;
impurity A =about loS; impurity G =about 1.6. HO 0 H H
System miUlbililY Reference solution (b):
- resolution: minimum 2.5 between the peaksdue to B. 4,Scr-epoxy-17-methyl-7,S-didehydromorphinan-3,6<x-diol
impurities F and H; minimum 1.5 between the peaks due (morphine),
to impurities D and A.
CH,
Calculation of percentage contents: H N
I
- comaion faaon: multiply the peak areas of the following

impurity C = 0.7; impurity G = 0.2; impurity 1= 1.3;
phosphate sesquihydrate in reference solution (a).
Limits:
- impun°'Y A: maximum 1.0 per cent;
- impurity H: maximum 0.25 per cent;
- impurities C, D, E: for each impurity, maximum Co 4,5a::4' ,5'a-diepoxy-3,3 /-dimethoxy-17,17 /-dimethyl-
0.2 per cent; 7)7',8,8' -retradehydrc-z.z"-bimorphinan-6a,6' e-diol
- impurities B, F, G, I: for each impurity, maximum (codeine dimec),
0.15 per cent;
0010 per cent;
Sulfates (2.4.13)
Dilute S mL of solution S to 20 mL with distilled warer R.
5.0 per cent [0 7.5 per cent, determined on 0.500 g by
drying in an oven at lOS "C.
ASSAY D. 4,Sa-epoxy-2-[(4,Sa-epoxy-6<x-hydroxy-17-methyl-7,S-
didehydromorphinan-3-yl)oxy]-3-methoxy-17-methyl-7,S-
Dissolve 00365 g in 50 mL of anhydrous acetic acid R. Titrate
didehydromorphinan-tic-ol (3-D-(codein-2-yl)morphine),
with 0.1 M perchlorie acid, determining the end-point
potentiome meally (2. 2.21f) .
I mL of 0.1 M perchloric acid is equivalent to 39.74 rng
of ClSH,¢W,P.
STORAGE
IMPURITIES
Speafiedimpurities A, BJ C, D, EJ F, GJ H, 1.
E. 4,Sa-epoxy-3-methoxy-l7-methyl-7,S-
dldehydromorphinan-ee.Iuc-diot,
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2022 Codergocrine Mesilate 1-657
,
CH,
H N
-~~
F. 4,SG<-epoxy-3-methoxy-17-methyl-7,8- L. 4,SG<-epoxy-6-methoxy-17-methyl-6,7,8,14-
didehydromorphinan-6cx,14-diol, tetradehydromorphinan-3-o1 (oripavine),
,
CH,
H3CO
~ i8=>
r_
H N
.•
o/H
-- ° -- \,.
OH
HO H...o
N H
\
OCH,
I
G. 4,SG<-epoxy-3,6-dimethoxy-17-methyl-6,7,8,14- CH,
.M.7J7'-oxybis(4,5et.-epoxy-3-methoxy-17-methyl-6,7,8,14-
tetradehydromorphinan-6-o1) (7,7'-oxybis(6-D-
~~
demethylthebainej).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
H3CO °HH
Codergocrine Mesilate ***

H. 4,SG<-epoxy-3-methoxy-7,8-didehydromorphinan-oo-ol *** ***
(norcodeine), (Ph. Bur. monograph 2060) ***
SO,"
• I
CH,
R
Name Mol. Formula Mr I
I. 4,SG<-epoxy-3-methoxy-17-methyl-7,8-
dlhydroergocomine
mesllale
<"H.,N,OsS 680
H 3C
yCH 3
dihydroergocrisline
mesllale
~~N508S 108
P CH,
u-dihydroergoayptioe ~~1N508S 61. ('CH'

meeuate
J. (17RS)-4,SG<-epoxy-oo-hydroxy-3-methoxy-17-methyl-7,8- CH,
didehydromorphinan 17-oxide (codeine N-oxide), p-dihyclroergocrypline
mesaete
CnH47 N!>OaS 61.
~'S(
8067-24-1
Action and use

Vasodilator.
PhE" _
K. 4,SG<-epoxy-14-hydroxy-3-methoxy-17-methyl-7,8- DEFINITION
didehydromorphinan-6-one (14-hydroxycodeinone), A mixture of:
- (6aR,9R,IOaR)-N-[(2R,SS,IOaS,IObS)-IOb-hydroxy-2,5-
bis(I-methylethyl)-3,6-dioxooClahydro-8H-oxazolo [3,2-01
pyrrolo[2,I-e1pyrazin-2-yl]-7-methyl-4,6,6a,7,8, 9,I0, I Oa-
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1-658 Codergocrine Mesilate 2022
oetahydroindolo[4,3-fg]quinoline-9<arboxamide principal spot in the chromatogram obtained with the

methanesulfonate (dihydroergocomine mesilate); reference solution.
- (6aR,9R, I OaR)-N-[(2R,5S, IOaS,I0bS)-5-benzyl-1 Ob- B. Examine the chromatograms obtained in the test for
hydroxy-2-(I-methylethyl)-3,6-dioxooetahydro-8H-oxazolo composition.
[3,2-a] pyrrolo[2,1-,] pyrazin-2-yl]-7-methyl- Results The 4 principal peaks in the chromatogram obtained
4,6,6.,7,8,9, I 0, IOa-octahydroindolo [4,3-fg] quinoline-9- with the test solution are similar in retention time to the
carboxamide methanesulfonate (dihydroergocristine 4 principal peaks in the chromatogram obtained with the
mesilate);
reference solution.
- (6aR,9R, I OaR)-N-[(2R,5S, lOaS,IObS)-1 Ob-hydroxy-2-(l-
methylethyl)-5-(2-methylpropyl)-3,6-dioxooetahydro-8H- TESTS
oxazolo[3,2-a]pyrrolo[2, 1-,] pyrazin-2-yl]- 7-methyl- pH (2.2.3)
4,6,6a,7,8, 9, 10, I Oa-octahydroindclo[4,3-jg] quinoline-9- 4.2 to 5.2.
carboxamide methanesulfonate (ce-dihydroergocryptine Dissolve 0.10 g in ,arbon dioxide-free waterR and dilute to
mesilate); 20 mL with the same solvent.
- (6aR,9R,IOaR)-N-[(2R,5S,IOaS,10bS)-IOb-hydroxy-2-(l- ComposItion
methylethyl)-5-[(1RS)-I-methylpropyl]-3,6- Liquid chromatography (2.2.29): use the normalisation
dioxooctahydro-8H-oxazolo[3,2-a]pyrrolo[2,1-,]pyrazin-2- procedure.
yl]-7-methyl-4,6,6a,7,8,9, I 0, 1Oa-oc tahydroindolo [4,3-fg]
quinoline-9-carboxamide methanesulfonate
examined in a mixture of 1 volume of anhydrous ethanol R
(p-dihydroergocryptine mesilate or epicriptine mesilate).
and 2 volumes of a 10 gIL solution of tartaric acid Rand
Content dilute to 10 mL with the same mixture of solvents.
Reference solution Dissolve 20 mg of codergoaine
PRODUCTION mesilate CRS in a mixture of I volume of anhydrous ethanol R
It is considered that alkylsulfonate esters are genotoxic and and 2 volumes of a 10 gIL solution of tartan', acid Rand
are potential impurities in codergocrine mesilate. dilute to 10 mL with the same mixture of solvents.
The manufacturing process should he developed taking into Column:
consideration the principles of quality risk management, ~ size: 1= 0.15 m, 0 = 4.6 nun;
together with considerations of the quality of starting - stationaryphase: octade<y/silyl silica gelfor chromatography R
materials, processcapability and validation. The general (5 urn).
methods 2.5.37. Methyl, ethyl and isopropyl metharwsulfonate in
Mobile phase tn'ethyfamine R, acetonitrile RJ water R
methanesuJfoni< acid, 2.5.38. Methyl, ethyl and isopropyl (2.5:25:75 VIVIV).
methanesulfonaze in acn've substances and
F/Qw rau 1.5 mUrnin.
2.5.39. Methanesulfonyl chloride in methanesulfoni, acid are
available to assistmanufacturers. Detection Spectrophotometer at 280 nm.
CHARACTERS Injection 20 1'1-.
Appearance Run time 20 min.
White or yellowish powder. Elution order Dihydroergocornine, c-dihydrcergccryptine,
Solublllty dihydroergocristine, p-dihydroergo,ryptine.
Sparingly soluble in water, sparingly soluble to soluble in System suitability Test solution:
ethanol (96 per cent), slightly soluble in methylene chloride. ~ resolution: minimum 3 between any 2 consecutive principal
peaks.
IDENTIFICATION
A. Thin-layer chromatography (2.2.27). Composition:
~ dihydroergocornine: 30.0 per cent to 35.0 per cent;
Test solution Dissolve 0.20 g of the substance to be - a-dihydroergocryptine: 20.0 per cent to 25.0 per cent;
examined in a mixture of 1 volume of methanol Rand - dihydroergocristine: 30.0 per cent to 35.0 per cent;
9 volumes of methylene chloride R and dilute to 5 mL with the - p-dihydroergocryptine: 10.0 per cent to 13.0 per cent,
same mixture of solvents. - disregard limit: 1.0 per cent.
Reference sol",wn Dissolve 0.20 g of methanesulfoni< acid R in
Related substances
a mixture of I volume of methanolRand 9 volumes of
Thin-layer chromatography (2.2.27). Perform the testas rapidly
methylene ,hloride R and dilute to 5 mL with the same
as possible and prouaed from dina light. Prepare the lestsolution
mixture of solvents.
las, and immediately before application on the plate.
Mobile phase water R, concentrated ammonia R, butanol R, examined in a mixture of I volume of methanol Rand
acetone R (5:10:20:65 VIVIVII'). 9 volumes of methylene chloride R and dilute to 5.0 mL with
Application 10 ~L. the same mixture of solvents.
Development Over 2/3 of the plate. Reference solution (aJ Dissolve 40 mg of dihydroergocristine
Drying In a current of cold airfor not more than 1 min. mesilate CRS in a mixture of I volumeof methanol Rand
Detection Spray with a I gIL solution of bromocresol purple R 9 volumes of methylene chloride R and dilute to 10.0 mL with
in methanol RJ adjusted to a violet-red colour with 0.05 mL the same mixtureof solvents. Dilute 3.0 mL of the solution
of diluu ammonia RJ. to 50.0 mL with a mixture of 1 volume of methanol R and
9 volumes of methylene chloride R.
Drying In a current of hot airat 100 "C.
Reference solution (b) To 2.0 mL of reference solution (a),
add 1.0 mL of a mixture of 1 volume of methanol Rand
with the test solutionis similar in positionand colour to the
9 volumes of methylene chloride R.
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2022 Cod-liver Oil 1-659
Reference solution (c) To 1.0 mL of reference solution (a), Content

add 2.0 mL of a mixture of 1 volume of methanolRand - sum 0/ the contents 0/EPA and DBA (expressed as
9 volumes of methylene chloride R. triglycerides): 10.0 per cent to 28.0 per cent;
Reference solution (d) To 1.0 mL of reference solution (a), - vitamin A: 50 ill (15 pg) [0 500 ill (150 ~g) per gram;
add 5.0 mL of a mixture of I volume of methanol Rand - vitamin D3 : maximum 50 IU (1.3 ug) per gram.
9 volumes of methylene chloride R. A suitable antioxidant may be added.
Piau TLC silicagel piau R. PRODUCTION
Mobilephase concentrated ammonia R, melhanolR) ethyl The fish shall only be given feed with a composition that is
acetauR, methylene chrome R (1:3:50:50 VIVIVIV). in accordance with the relevant European Union or other
Application I0 ~L. applicable regulations.
Drying In the dark for 2 min after the application of the last The content of dioxins and dioxin-like PCBs
solution. (polychlorinated biphenyls) is controlled using methods and
First development In an unsaturated tank, over 2/3 of the limits in accordance with the requirements set in the
plate. European Union or other applicable regulations.
Drying In a current of cold air for not more than 1 min. CHARACTERS
Second development In an unsaturated tank, over 2/3 of the Appearance
plate; use freshly prepared mobile phase. Clear, pale yellowish liquid.
Drying In a current of cold air for not more than J min. Solubility
Practically insoluble in water, miscible with light petroleum,
Detection Spray thoroughly with dimezhy/aminobenzaldehyde
solution R 7 and dry in a current of hot air until the spot in slightly soluble in ethanol (96 per cent).
the chromatogram obtained with reference solution (d) is IDENTIFICATION
clearly visible. A. Examine the 13C NMR spectra obtained in the test for
System suitability Test solution: positional distribution (~(2)-acyJ) offatty acids (see Tests).
- the chromatogram shows at least 3 separated secondary The spectra contain peaks between 172 ppm and 173 ppm
spots. with shifts similar to those in the spectrum shown in
Limits: Figure 2398.-1.
- any impurity: any spots, apart from the principal spot, are The positional distribution (~(2)-acyl) for cervonic
not more intense than the spot in the chromatogram (docosahexaenoic) acid (C22:6 n-3; DHA), timnodonic
obtained with reference solution (a) (0.3 per cent); not (eicosapentaenoic) acid (C20:5 n-3; EPA) and moroctic acid
more than 4 such spots are more intense than the spot in (CI8:4 n-3) complies with the limits.
the chromatogram obtained with reference solution (e) B. Linoleic acid (see Tests).
(0.1 per cent) and 2 of these may be more intense than
TESTS
the spot in the chromatogram obtained with reference
Acid value (2.5.1)
Maximum 2.0.
Anisidine value (2.5.36)
Maximum 5.0 per cent, determined on 0.500 g by drying at
Maximum 10.0.
120 'C under high vacuum.
Peroxide value (2.5.5, M"hod B)
ASSAY Maximum 5.0.
Dissolve 0.500 g in 60 mL of pyridine R. Pass a stream of
nitrogen R over the surface of the solution and titrate with Unsaponifiable matter (2.5.7)
0.1 M tetrabutylamrnonium hydroxide, determining the Maximum 1.5 per cent, determined on 2.0 g, and extracting
end-point potentiometrically (2.2. 2ff). with 3 quantities, each of 50 mL, of peroxide--free ether R.
1 mL of 0.1 M tetrabutylarnrnonium hydroxide is equivalent Stearin
to 68.04 mg of codergocrine mesilate (average Me = 680). Heat at least 10 mL to 60-90 °C then allow to cool for 3 h
in a bath of iced water or a thennostatically controlled bath
STORAGE at 0 ± 0.5 °C. If necessary, to eliminate insoluble matter,
Protected from light. filter the sample after heating. The sample remains clear.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Positional distribution (~(2)-acyl) of fatty acids
Nuclear magnetic resonance spectrometry (2.2.33).
Test solution Dissolve 190-210 mg of the substance to be
examined in 500 ul, of deuterated chloroform R. Prepare at
Farmed Cod-liver Oil least 3 samples and examine within 3 days.
Apparatus High-resolution Ff-NMR spectrometer
operating at minimum 300 MHz.
Action and use Acquisition 0/ "c NMR spectra The following parameters
Source of vitamins A and D. may be used:
PhE" ~ _ - sweep width: 200 ppm (-5 ppm to 195 ppm);
- irradiation frequency offset: 95 ppm;
DEFINITION - time domain: 64 K;
Purified fatty oil obtained from the fresh livers offanned cod, - pulse delay: 2 s;
Gadus morhua L., solid substances being removed by cooling - pulseprogram: zgig 30 (inverse gated, 30° excitation pulse);
and filtering. - dummy scans: 4;
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1-660 Cod-liver Oil 2022
2
4
173.4 173,2 173.0 172.8 172.6 172.4 172.2 172.0 171.8 ppm
I. Cl C18:4 2. a: EPA 3. ~ C18:4 4. PEPA 5.aDHA
Figure 2398.-1. - BC NMR spectrum: carbonyl region offanned cod-liver oil
- number of scans: 4096. IOOxp

Processing andplotting The following parameters maybe " +P
used: a peakarea of the corresponding a-carbonyl peak;
- size: 64 K (zero-filling); fJ peak area of p-carbonyl peakfrom C22:6 n-j, C20:5 0-3 or
- window multiplkation: exponential; C18:4 n-S, respectively.
- Lorenteian broadening faaor: 0.2 Hz.
Use the CDClj signal for shift referencing. The shift of the Limits:
central peak of the I: I: I triplet is set to 77 .16 ppm. - positional distribution (P(2)-acyl):
- eervonie (docosahexoenoic) acid (C22:6 n-3; DHA):
Plot the speetral region B 171.5-173.5 ppm. Compare the 71 per cent to 81 per cent;
spectrum with the spectrum shown in Figure 2398.-1. - timnodonie (eirosapentaenoic) acid (C20:5 n-3 EPA):
The shift values lie within the ranges given in Table 2398.-1.
32 per cent to 40 per cent;
Table 2398.-1. - Shift values - mOTOCtk acid (C18:4 n-3): 28 per cent to 38 per cent.
Signal ShIft range (ppm) Composltlon offatty acids (2.4.29)

For identification of the peaks, see the chromatogram shown
pDHA 172.05- 172.09
172.43 . 172.47
in Figure 2398.-2.
.DHA
PEPA 172.52 - 172.56 The 24 largest peaks of the methyl esters account for more
a EPA 172.90 - 172.94 than 90 per cent of the total area (thesecorrespond to, in
p CIS:. 172.56 - 172.60 common elution order: 14:0, 15:0, 16:0, 16:1 0-7, 16:40-1,
Cl C18;4 172.95·172.99 18:0,18:1 n-c, 18:1 n-7, 18:2 n-6, 18:3 n-3, 18:4 n-3,
20:1 n-II, 20:1 n-c, 20:1 n-7, 20:2 n-6, 20:4 n-6, 20:3 n-3,
20:4 n-3, 20:5 n-3, 22:1 n-II, 22:1 n-9, 21:5 n-S, 22:5 n-3,
System suitability:
22:6 n-3).
- signal-to-noise ratio: minimum 5 for the smallest relevant
peak corresponding to " C 18:4 signal (in the Linoleic acid (2.4.29)
range B 172.95-172.99 ppm); 3.0 per cent to 11.0 per cent.
- peak width at hal}heighr. maximum 0.02 ppm for the ASSAY
central CDCI, signal (at B 77.16 ppm). EPA and DBA (2.4.29)
Calculation of positional distribution (P(2)-acyO Use the See the chromatogram shown in Figure 2398.-2.
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2022 Cod-liver Oil 1-661
3 7
--=::
~
::
~
~ 24
-:::= 19
--=
~
~
~ 4
9
1 1
~
~
~
-:::
-= 6
--=::
::: 13
....::
1
~
~
1011 20 23
1
--=
}
~=
11,j
en
U
l!f1Ill~I' !~1I11
i I
,II.
" ,
4 \5
, " ,.
16 ,1f
'117 21
,
)2
I II
". I'll;
,
II 1II1I 1" 111 II II I' 1111"111"11111"1""111"1' 11111111111"111"111111""1111111"'1'1111'1"111'11""1"'
6 8 10 12 14 16 18 20 22 24 26 min
1. CJ4:0 5. G16:4 n-I 9. C18:2 n-6 13. e20:! 0-9 17. C20:3 0-3 21. e22:1 0-9
2. C15:0 6. C18:0 to CIS:3 0-3 14. C20:1 0-7 18. C20:4 0-3 22. C21:5 0-3
3. C16:0 7. CIS:! 0-9 11. C18:4 0-3 15. C20:2 0-6 19. C20:5 n-S 23. C22:5 n- J
4. GI6:) n-7 8. C18:1 0-7 12. C20:1 n-II 16. e20:4 n-6 20. e22:l 0-11 24. C22:6 0-3
Figure 2398.-2. - Chromawgram for the USt for composition of fatlY acids of fanned cod-liver oil
Vitamin A matched 1 em cells, using 2-propano/ RJ as the compensation

Cany out the test as rapidly as possible, avoiding exposure to liquid.
actinic lightand air, oxidising agents, oxidation catalysts (for Calculate the content of vitamin AJ as aU-trans-retinol, in
example, copper and iron) and adds. International Units per gram, using the following expression:
Use method A. If method A is found not to be valid, use
method B. 1821
A 325 x - - X V
METHOD A 100m
Ultraviolet absorption spectrophotometry (2.2.25). Aj 2 3" absorbance at 325 nm;
Testsolution To 1.00 g in a round-bottomed flask, add m mass of LIte substance 10be examined, in grams;
V "total volume of solution containing 11).15 IV of vitamin A per
3 mL of a freshly prepared 50 per cent mlm solution of rnillilitre;
potassium hydroxide Rand 30 mL of anhydrous ethanol R. Boil 1821 conversionfactor for the specific absorbanceof aU-tnJJIS-retinol,
under reflux in a current of nitrogen R for 30 min. Cool in International Units.
rapidly and add 30 mL of waterR. Extract with 50 mL of
ether R. Repeat the extraction 3 times and discard the lower The above expression can be used only if A 325 has a value
layer after complete separation. Wash the combined upper not greater than A 3 25 ,corr 10.970, where A 325,oorr is the
layers with 4 quantities, each of 50 mL, of water R, and corrected absorbance at 325 run and is given by the following
evaporate to dryness under a gentle current of nitrogen R at a equation:
temperature not exceeding 30°C or in a rotary evaporator at
a temperature not exceeding 30 °C under reduced pressure A 325 , <orr = 6.815A 325 - 2.555A 3I o - 4.260A 334
(water ejector). Dissolve the residue in sufficient
2-propanol Rl to give an expected concentration of vitamin A A designates the absorbance at the wavelength indicated by
equivalent to 10-15 IV/mL. the subscript.
Measure the absorbances of the solution at 300 run, 310 nrn, If A 32 5 has a value greater than A325,corr 10.970, calculate the
325 om and 334 om and at the wavelength of maximum content of vitamin A using the following expression:
absorption with a suitable spectrophotometer in specially
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1-662 Cod-liver Oil 2022
1821
A 325 ccrr X - - X V The exact concentration of reference solution (b) is assessed
. 100m by ultraviolet absorption spectrophotometry (2.2.25). Dilute
reference solution (b) with 2-propanol Rl to a presumed aU-
The assay is not valid unless:
trans-retinol concentration of 10-15 IU/mL and measure the
- the wavelength of maximum absorption lies between
absorbance at 325 om in matched 1 ern cells using
323 nrn and 327 nrn;
2-propano/ RJ as the compensation liquid.
- the absorbance at 300 nm relative to that at 325 om is at
most 0.73. Calculate the content of all-trans-retinol in International
Units per millilitre of reference solution (b), using the
METHODB following expression:
Test solution Prepare duplicates. To 2.00 g in a round- A 1821 x V,
bottomed flask, add 5 mL of a freshly prepared 100 gIL 325 X 100x V4
solution of ascorbic add R, 10 mL of a freshly prepared
A J2 5 absorbance at 325 noli
800 gIL solution of potassium hydroxide R and 100 mL of VJ volume of the diluted solution;
anhydrous ethanol R. Boil under a refluxcondenser on a V"' volume of reference solution (b) used;
water-bath for 15 min. Add 100 mL ofa 10 gIL solution of l82l conversion factor for the specific absorbance of all-ln1ll$-relinol,
in International Units.
sodium chloride R and cool. Transfer the solution to a 500 mL
separating funnel, rinsing the round-bonomed flask with
Column:
about 75 mL ofa 10 gIL solution of sodium chloride Rand
then with 150 mL of a mixture of equal volumes of ether R
- size: I = 0.25 m, 0 = 4.6 mm;
- stationary phase: ocrade<y/silyl SIlica gel for chromatography R
and ligh. petroleum RI. Shake for 1 min. When the layers
(5-10 pm),
have separated completely, discard the lower layer and wash
the upper layer, first with 50 mL of a 30 gIL solution of Mobile phase water R, me<hanol R (3:97 VIV).
potassium hydroxide R in a 10 per cent VIV solution of Flow rate 1 ml.Jmin.
anhydrous ethanol R and then wilh 3 quantities, each of Detection Spectrophotometer at 325 om.
50 mL, of a 10 gIL solution of sodium chloride R. Filter the Injection 10 f.tL; inject in triplicate the test solution and
upper layer through 5 g of anhydrous sodium sulfate R on a reference solution (b).
fast filter paper into a 250 mL flask suitable for a rotary
evaporator. Wash the funnel with 10 mL of fresh extraction
Retention time All-trans-retinol = 5 ± 1 min.
mixture, filter and combine the upper layers. Distil them at a System suitability:
temperature not exceeding 30 °C under reduced pressure - the chromatogram obtained with the test solution shows a
(water ejector) and fill with nitrogen R when evaporation is peak corresponding to the peak due to aU-trans-retinol in
completed. Alternatively, evaporate the solvent under a gentle the chromatogram obtained with reference solution (b);
current of nitrogen R at a temperature not exceeding 30 °C. - the results obtained with the duplicate test solutions do
Dissolve the residue in 2-propanol R, transfer to a 25 mL not differ by more than 5 per cent;
volumetric flask and dilure to 25 mL with 2-propanol R. - the recovery of aU-tram-retinol in reference solution (b) as
Gentle heating in an ultrasonic bath may be required. A large assessed by direct absorption spectrophotometry is greater
fraction of the while residue is cholesterol, constituting than 95 per cent.
approximately 50 per cent mlm of the unsaponifiable matterof Calculate the content of vitamin A using the following
cod-liver 00. expression:
Reference solution (a) Prepare a solution of retinol Cx V I
acetate CRS in 2-propanol Rl so that 1 mL contains about A,x--x-
1000 ill of aU-trans-retinol.
A2 m
The exact concentration of reference solution (a) is assessed Al area of the peak due to aU-ln1m-retinol in the chromatogram
obtained with the lest solution;
by ultraviolet absorption spectrophotometry (2.2.25). Dilute
A2 area of the peak due to aU-ln1ns-retinol in the chromatogram
reference solution (a) with 2-propanol RJ to a presumed obtained with reference solution (b);
concentration of 10-15 IU/mL and measure the absorbance C concentration of retinolaceroll CRS in reference solution (a) as
at 326 nm in matched 1 em cells using 2-propanol Rl as the assessed prior to the saponifical:ion, in International Units per
milliliue (= 1000 IU/mL);
compensation liquid.
V volume of reference solution (a) treated (2.00 mL);
Calculate the content of vitamin A in International Units per nI mass of the substance to be examined in the test solution
millilitre of reference solution (a) using the foUowing (2.00 g).
expression, taking into account the assigned content of retinol
autate CRS: Vitamin D,
Liquid chromatography (2.2.29). Cany OIl' <he assayas rapidly
A 1900x V2 as possible, avoiding exposure UJ aainic lightand air.
X
326 100x VL lmemal standard solution Dissolve 0.50 mg of
ergocaki/erol CRS in 100 mL of anhydrous e<hanol R.
A J26 absorbance at 326 noli
VI volume of reference solution (a) used; Testsolution (aJ To 4.00 g in a round-bottomed flask, add
V2 volume of the diluted solution; 5 mL of a freshly prepared 100 gIL solution of ascorbic
1900 convenion factor for the specific absorbance of retinol acid R, 10 mL of a freshly prepared 800 gIL solution of
aUUJ.le CRS, in International Units. potassium hydroxide R and 100 mL of anhydrous ethanol R.
Boil under a reflux condenser on a water-bath for 30 min.
Reference solution (b) Proceed as described for the test
Add 100 mL of a 10 gIL solution of sodium chloride Rand
solution but using 2.00 mL of reference solution (a) in place
cool the solution to room temperature. Transfer the solution
of the substance to be examined.
to a 500 mL separating funnel, rinsing rhe round-bottomed
flask with about 75 mL of a 10 gIL solution of sodium
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2022 Cod-liver Oil (Type A) 1-663
chloride R and then with 150 mL of a mixture of equal Calculate the content of vitamin D'l in International Units
volumes of ether R and light petroleum R1. Shake for 1 min. per gram using the following expression, taking into account
When the layers have separated completely, discard the lower the assigned content of cholecalciferol CRS;
layer and wash the upper layer, first with 50 mL of a 30 gIL
solution of potassium hydroxide R in a 10 per cent VfV Az A) m2 Vz 4
-x x-x-xO
solution of anhydrous ethanol R, and then with 3 quantities, A6 A [As] A
4- - X 2
mL VI
each of 50 rnl., of a 10 gIL solution of sodium chloride R. Al
Filter the upper layer through 5 g of anhydrous sodium
ml mass of the sample in test solution (b), in grams]
sulfate R on a fast filter paper into a 250 mL flask suitable for mz total mass of chol,,",kiJerol eNS used for the preparation of
a rotary evaporator. Wash the funnel with 10 mL of fresh reference solution (a), in micrograms (500 Il&l;
extraction mixture, filter and combine the upper layers. Distil AI area (or height) of the peak due to cholecalciferol in the
them at a temperature not exceeding 30 °C under reduced chromatogram obtained with lest solution (a);
pressure (water ejector) and fill with nitrogen R when A, area (or height) of the peak due to cholecalciferol in the
chromatogram obtained with test solution (b);
evaporation is completed. Alternatively, evaporate the solvent Aj area (or heighl) of the peak due 10 ergocalciferol in the
under a gentle current of nitrogen R at a temperature not chromatogram obtained wi!h reference solution (b);
exceeding 30 "C. Dissolve the residue in 1.5 mL of the At area (or height) of the peak due to ergocalciferol in the
mobile phase described under Purification. Gentle heating in chromatogram obtained with lest solution (b);
A~ area (or height) of a possible peak in the chromatogram
an ultrasonic bath may be required. A large fraction of the obtained with test solution (a) with the same retention time as
white residue is cholesterol, constituting approximauly 50 per cent lhe peak co-eluting with ergocalciferol in test solution (b);
mlm of the unsafH»lijiabfe matterof cod-lwer oil. A6 area (or height) of the peak due 10 cholecalciferol in the
Test solution (b) Prepare duplicates. To 4.00 g add 2.0 mL VI total volume of reference solution (a) (lOll mL)j
of the internal standard solution and proceed as described for V, volume of reference solution (a) used for preparing reference
test solution (a). solution (b) (2.0 mL).
Reference solution (a) Dissolve 0.50 mg of cholecalciferol CRS
in 100.0 mL of anhydrous ethanol R. STORAGE
Reference solution (b) In a round-bottomed flask, add In an airtight and well-filled container, protected from light
2.0 mL of reference solution (a) and 2.0 mL of the internal If no antioxidant is added, store under an inert gas.
standard solution and proceed as described for test Once the container has been opened, its contents are used as
solution (a). soon as possible and any part of the contents not used at
PURIFICATION once is protected by an atmosphere of inert gas.
Column: LABELLING
- size: I ;;;; 0.25 rn, 0 ;;;; 4.6 mm; The label states:
- swtil)1lary phase: nitrile silica gel for chromatography R - the concentration of EPA and DHA, expressed as
(10 urn). triglycerides;
Mobile phase isoamyl alcohol R, hexane R (1.6:98.4 VIII). - the number of International Units of vitamin A per gram;
Flow Tau 1.1 mUmin. - the number of International Units of vitamin D, per
gram.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIlE"
Injection 350 ~L of reference solution (b) and test
solutions (a) and (b). Collect each eluate from 2 min before
until 2 min after the retention time of cholecalciferol, in a
ground-glass-stoppered tube containing I mL of a I gIL
solution of butyihydroxytoluene R in hexane R. Evaporate Cod-liver Oil (Type A)
separately to dryness at a temperature not exceeding 30 °C
under a gentle current of nitrogen R. Dissolve each residue in (Ph. Bur. monograph 1192)
1.5 mL of acetonitrik R. Action and use
DETERMINATION Source of vitamins A and D.
Column: Each IV ofvitamin D, is equivalent to 0.025 ug of
=
- size: l 0.15 m, 0;;;; 4.6 mm; colecalciferol.
- stationary phase: octadecylsi/yl silica gelfor chromatography R PIlElr _
(5 IlID).
Mobile phase phosphoric acidR, 96 per cent VW solution of DEFINITION
acetonitrile R (0.2:99.8 VIV). Purified fatty oil obtained from the fresh livers of wild cod,
Flow rate 1.0 mUmin. Gadusmorhua L. and other species of Gadidae, solid
substances being removed by cooling and filtering. A suitable
antioxidant may be added.
Injeaion 2 quantities not exceeding 200 j!L of each of the
Content
3 solutions obtained under Purification.
- oitamin A: 600 IU (180 Ilg) to 2500 IU (750 Ilg) per
Systemsuiwbility; gram;
- resolution: minimum 1.4 between the peaks due to - vitamin D 3 : 60 IV (1.5 ~g) to 250 ill (6.25 ug) per gram.
ergocalciferol and cholecalciferol in the chromatogram
obtained with reference solution (b); PRODUCTION
- the results obtained with the test solution (b) duplicates The content of dioxins and dioxin-like PCBs
do not differ by more than 5 per cent. (polychlorinated biphenyls) is controlled using methods and
limits in accordance with the requirements set in the
European Union or other applicable regulations.
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1-664 Cod-liver Oil (Type A) 2022
CHARACTERS Trivial name of fatty Nomenclature Lower llmlt Upper llmit

acid area (per cent) area
Appearance (per cent)
Clear, yellowish liquid.
Samrottd faeryacids:
Solubility Myristic acid 14:0 2.0 6.0
Practically insoluble in water, miscible with light petroleum, Palmitic acid 16:0 7.0 14.0
slightly soluble in ethanol (96 per cent). Stearic acid 18:0 1.0 4.0
MOIIQ-lUlSalUTattd Jauy acids:
IDENTIFICATION
Palmitoleic acid 16:10·7 4.5 11.5
First identification: A J B.. C. cs-vaccenk acid 18:1 0·7 2.0 7.0
Second idenrificalion: C.. D. Oleic acid 18:1 n-9 12.0 21.0
llidoleic acid. 20:1 n-l l 1.0 5.5
A. In the assay for vitamin A using method A, the test Gondoic acid 20:1 n-s 5.0 17.0
solution shows an absorption maximum (2.2.25) at Erucic acid 22:1 0-9 0 1.5
325 ± 2 run. In the assay for vitamin A using method B, the Cetcleic acid (22:1 22:1 n-Il+13 5.0 12.0
chromatogram obtained with the test solution shows a peak 0-11)
corresponding to the peak due to aU-trans-retinol in the PoIy-unsaturaudJatry acds:
chromatogram obtained with the reference solution. Linoleic acid 18:2 n-6 0.5 3.0
«-Lieolenic acid 18:30-3 0 2.0
B. In the assay for vitamin D 3) the chromatogram obtained MOI'ocOc acid 18:40·3 0.5 4.5
with test solution (a) shows a peak corresponding to the peak Tinmodonic 20:50-3 7.0 16.0
due to cholecalciferol in the chromatogram obtained with (ekosapentaenolc) acid
reference solution (b). (EPA)
Cersonic 22:60-3 6.0 18.0
(docOsahexaenoic) acid
D. To 0.1 g add 0.5 mL of metll)lfene chloride R and I mL of (DHAl
antimony trichWride solution R. Mix. A deep blue colour
develops in about lOs. Test solution Dilute about 0.45 g of the substance to be
TESTS examined (0 10.0 mL with a 50 mWI.solution of
Appearance butyihydroxytoluene R in m'methy/pentane R. Transfer 2.0 mL
The substance to be examined is not more intensely coloured of this solution to a quartz tube and evaporate the solvent
than a reference solution prepared as follows: to 3.0 mL of with a gentle current of nitrogen R. Add 1.5 mL of a 20 WI.
red primary solution add 25.0 mL of yellow primary solution solution of sodium hydroxide R in methanol R, cover with
and dilute (0 50.0 mL with a 10 WI. solution of hydrochloric nitrogen R, cap tightly with a polytetrafluoroethylene-lined
acid R (2.2.2, Method II). cap, mix and heat on a water-bath for 7 min. Cool) add
2 mL of boron trichlorid~methanol solUlwn R, cover with
nitrogen R, cap tightly, mix and heat on a water-bath for
0.917 to 0.930.
30 min. Cool to 40-50 °C, add I mL of m'methy!pentane R,
Refractive index (2.2.6) cap and vortex or shake vigorously for at least 30 s.
1.477 to 1.484. Immediately add 5 mL of saturatulsodium chloride solution R,
Acid value (2.5.1) cover with nitrogen R, cap and vortex or shake vigorously for
Maximum 2.0, determined on 20.00 g. at least 15 s. Allow the upper layer to become clear and
Anisld1ne value (2.5.36) transfer it to a separate tube. Shake the methanol layer once
Maximum 30.0. more with I mL of trimethy!pentane R and combine the
trimel:hylpentane extracts. Wash the combined extracts with
Iodine value (2.5.4, Method B) 2 quantities, each of 1 mL, of water R and dry over anhydrous
150 to 180, determined on 0.14 g to 0.16 g. sodium sulfate R. Prepare 2 solutions for each sample.
Use starch solution R2. Reference solution Dissolve 0.300 g of methylarachidate R,
Peroxide value (2.5.5, Method B) 0.300 g of metll)ll behenau R, 0.300 g of metll)ll palmitote R
Maximum 10.0. and 0.300 g of metlryl stearaU R in a 50 mWI.solution of
Unsaponifiable matter (2.5.7) butyihydroxytoluene R in trimethylpentane R and dilute to
Maximum 1.5 per cent, determined on 5.0 g. 10.0 mL with the same solution.
Stearin Column:
Heat at least 10 mL to 60-90 °C and cool to room - material: fused silica;
temperature. Immerse for 3 h in a bath of iced water or a - size: 1== 30 rn, 0 = 0.25 mm;
thennostaticaUy controlled bath at 0 ± 0.5 °C. If necessary, - stationary phase: macrogol 20 000 R (film thickness
to eliminate insoluble matter, filter the sample after heating. 0.25 pm).
The sample remains clear. Canier gas hydrogen for chromatograplry R or helium for
chromatography R, with an oxygen scrubber.
Composition of fatty acids
Split ratio 1:200.
Temperature:
Time Temperature
(min) ("C)
Column 0-55 170 ..... 225
55 - 75 225
lniecticn port 250
Detector 2SO
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10 20 30 40 50 60 min
Figure 1192.-1. ~ Chromatogram for the testfor composition 01latty acids 01cod-/iver oil (rypt A)
Detection Flame ionisation. - the 24 largest peaks of the methyl esters account for more
Inj«tion I ~L, twice. than 90 per cent of the total area (these correspond to, in
common elution order: 14:0, 15:0, 16:0, 16:1 n-7,
System suitability:
16:4 n-L, 18:0, 18:1 n-9, 18:1 n-7, 18:2 n-6, 18:3 n-3,
- the 15 fatty acids to be tested are satisfactorily identified
18:4 n-3, 20:1 n-lI, 20:1 n-9, 20:1 n-7, 20:2 n-6, 20:4
from the chromatogram shown in Figure 1192.-1;
n-6, 20:3 0-3, 20:4 n-3, 20:5 n-3, 22:1 n-lI, 22:1 n-c,
- in the chromatogram obtained with the reference solution,
21:5 n-3, 22:5 n-3, 22:6 n-3).
multiply the areas of the peaks due to methyl palmitate,
methyl stearate, methyl arachidate and methyl behenate ASSAY
by the corresponding response factors in Table 1192.-1; VltamlnA
normalise the corrected areas of the peaks due to the fatty Cany out the test as rapidly as possible} avoiding exposure to
acid methyl esters to a sum of 100 per cent; the actinit. lightand air} oxidising agents, oxidation catalysts (for
normalised area percentage of each fany acid methyl ester example, copper and iron) and acids.
is to be within ± 1.0 percentage unit of the Use method A. If method A is found not to be valid, use
corresponding weight percentage; method B.
Table 1192.-1. MBTHODA
Ultraviolet absorption spectrophotometry (2.2.25).
Fatty acid methyl ester Theoretical response factor
Melbyl palmitate 1.049
Test solution To 1.00 g in a round-bottomed flask, add
Methyl stearate 1.029
3 mL of a freshly prepared 50 per cent mlm solution of
Methyl arachldate 1.013
potassium hydroxide Rand 30 mL of anhydrous ethanol R. Boil
under reflux in a current of nitrogen R for 30 min, Cool
Methyl benenate 1.000
rapidly and add 30 mL of water R. Extract with 50 mL of
- resolution: in the chromatogram obtained with. the test ether R. Repeat the extraction 3 times and discard the lower
layer after complete separation. Wash the combined upper
solution:
layers with 4: quantities, each of 50 ml., of water R, and
- minimum 1.3 between the peaks due to methyl oleate
evaporate to dryness under a gentle current of nitrogen R at a
and methyl es-vaccenare; the resolution between the
temperature not exceeding 30°C or by other suitable means
peaks due to methyl gadoleate and methyl gondoate is
at a temperature not exceeding 30 DC WIder reduced
sufficient for purposes of identification and area
pressure. Dissolve the residue in sufficient 2-propalWl RI to
measurement.
give an expected concentration of vitamin A equivalent to
Calculate the area per cent for each fatty acid methyl ester 10-15 IUlmL.
using the following expression:
Measure the absorbances of the solution at 300 nm, 310 om)
A, 325 run and 334 run and at the wavelength of maximum
A, x 100 absorption with a suitable spectrophotometer in specially
matched 1 em cells, using 2-propanol RJ as the compensation
A" peak area of Iarry acid Xj
A, swn of the peak areas (up to C22:6 n-3).
liquid.
Calculate the content of vitamin A, as ajl-ecas-retincl, in
The calculation is not valid unless: International Units per gram, using the following expression:
- the total area is based only on peaks due solely to fatty
/821
acid methyl esters; A 325 X -00 x V
- the number of fatty acid methyl ester peaks exceeding I "'
0.05 per cent of the total area is at least 24; An, absorbance at 325 nntj
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1-666 Cod-liver Oil (Type A) 2022
". mass of me substance to be examined. in grams; expression, taking into account the assigned content of retinol
V total volume ofsolulion containing 10-15 IU of vitamin A per
milhlitre;
acelate CRS:
1821 conversion factor for me specific absorbance ofall-mms-rerinol.
A 1900x V,
X
326 100xVI
The above expression can be used only if A 32 5 has a value
A l16 absorbance at 326 nm;
not greater than A 3 2 5,corIO.970) where A 3 2 5 ,COIT is the
VI volume of reference solution (a) used;
corrected absorbance at 325 run and is given by the following V2 volume of the diluted solution;
equation: 1900 conversion factor for the specific absorbance of retillol
acetare CRS, in International Units.
A"',mIT = 6.815A", - 2.555A 31O - 4,260A",
A designates the absorbance at the wavelength indicated by solution but using 2,00 mL of reference solution (a) in place
the subscript.
of the substance to be examined.
If A 325 has a value greater than A 325,COrr!O.970J calculate the
The exact concentration of reference solution (b) is assessed
content of vitamin A using the following expression:
1821 reference solution (b) with 2-propanof Rl to a presumed all-
A325 ,corr x 100m x V trans-retinol concentration of 10-15 IV/mL and measure the
absorbance at 325 run in matched 1 em cells using
The assay is not valid unless: 2-propanol Rl as the compensation liquid.
- the wavelength of the maximum absorption lies between
Calculate the content of aU-trans-retinol in International
323 nrn and 327 nrn;
Units per millilitre of reference solution (b), using the
- the absorbance at 300 urn relative to that at 325 om is at
most 0.73.
METHODB A 1821 X V,
X
Liquid chromatography (2.2.29). 325 100xV4
Testsolution Prepare duplicates. To 2.00 g of the substance An' absorbance at 325 run;
to be examined in a round-bottomed flask, add 5 mL of a V) volume of the diluted solution;
fresWy prepared 100 f'IL solution of aswrbic acid R, 10 mL of V~ volwne of reference solution (b) used;
a freshly prepared 800 f'IL solution of potassium hydroxitkR 1821 conversion factor for the specific absorbance of all-trons-relinol,
and 100 mL of anhydrous ethanolR. Boil under a reflux
condenser on a water-bath' for 15 min. Add 100 mL of a
Column:
10 gIL solution of sodium chloride R and cool. Transfer the
- size: 1= 0.25 m, 0 = 4.6 mm;
solution to a 500 mL separating funnel, rinsing the round-
- stationary phase: lXtadecyfsifyl silica gelfor chromalagraphy R
bottomed flask with about 75 mL of a 10 f'IL solution of
(5-10 um),
sodium chloride R and then with 150 mL of a mixture of
equal volumes of ether R and lightpetroleum RI. Shake for Mobile phase waterfor chromawgraphy R, methanol R
1 min. When the layers have separated completely, discard (3:97 VIV).
me lower layer and wash the upper layer, first with 50 mL of Flow rate 1 mIlrnin,
a 30 f'IL solution of potassium hydroxitk R in a Detection Spectrophotometer at 325 run.
10 per cent VIV solution of anlrydrous ethanol R and then Injection 10 f.lL; inject in triplicate the test solution and
with 3 quantities, each of 50 mL, of a 10 f'IL solution of reference solution (b).
sodium chloride R. Filter the upper layer through 5 g of
anhydrous sodium sulfate R on a fast filter paper into a Retention time All-trans-retinol = 5 ± 1 min.
250 mL flask. Wash the funnel with 10 mL of fresh System suitability:
extraction mixture, filter and combine the upper layers. Distil - the chromatogram obtained with the test solution shows a
them at a temperature not exceeding 30°C under reduced peak corresponding to the peak due to all-tram-retinol in
pressure and fill with nitrogen R when evaporation is the chromatogram obtained with reference solution (b);
completed. Alternatively, evaporate the solvent under a gentle - the results obtained with the duplicate test solutions do
current of nitrogen R at a temperature not exceeding 30°C. not differ by more than 5 per cent,
Dissolve the residue in 2-propanol RI, transfer to a 25 mL - the recovery of all-tram-retinol in reference solution (b) as
volumetric flask and dilute to 25 mL with 2-propalWl RI. assessed by direct absorption spectrophotometry is greater
Gentle heating in an ultrasonic bath may be required. A farge than 95 per cent.
fraction of the white residue is cholesterol, constituting Calculate the content of vitamin A using the following
approximately 50 per C<1lt mlm of the unsaponijiabfe matterof expression:
cod-liver oil
Cx V 1
Reference solution (a) Prepare a solution of retinol A,x--x-
acetate CRS in 2-propanol RJ so that 1 mL contains about A, m
1000 IV of all-trans-retinol. AI area of the peak due to aU-trons-relinoi in the chromatogram
The exact concentration of reference solution (a) is assessed obtained with the test solution;
A2 area of the peak due to all-trant-retinol in the chromatogram
reference solution (a) with 2-propanol RJ to a presumed C concentration of retind acetateCRS in reference solution (a) as
concentration of 10-15 IV/mL and measure the absorbance assessed prior to the saponification, in International Units per
at 326 run in matched 1 em cells using 2-propanol Rl as the millilitre (= 1000 JU/mL);
compensation liquid. V volume of reference solution (a) treated (2,00 mL);
m mass of the substance to be examined. in the test solution
Calculate the content of vitamin A in International Units per ('.00 g).
millilitre of reference solution (a) using the following
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Vitamin D 3 DETERMINATION
Liquid chromatography (2.2.29). Carty out the assqyas rapidly Column:
as possible, avoiding exposure to actinic lightand air. - size: 1= 0.15 m, 0 = 4.6 mm;
Internalstandard solution Dissolve 0.50 mg of - stationary phase: octadecylsilyl silica gelfor chromatography R
ergocalciferol CRS in anhydrous ethanol R and dilute to (5 urn),
100.0 mL with the same solvent. Mobile phase phosphoric acid R) 96 per cent VIV solution of
Testsolution (a) To 4.00 g of the substance to be examined acetonitrile R (0.2:99.8 VII').
in a round-bottomed flask, add 5 mL of a freshly prepared Flow rate 1.0 mllmin.
100 gIL solution of ascorbic acid R, 10 mL of a freshly Detection Spectrophotometer at 265 om.
prepared 800 gIL solution of potassium hydroxiik Rand
Injection 2 quantities not exceeding 200 J1L of each of the
100 mL of anhydrous ethanolR. Boil under a reflux
3 solutions obtained under Purification.
condenser on a water-bath for 30 min. Add 100 mL of a
10 gIL solution of sodium chloride R and cool the solution to Sys..m suitability:
room temperature. Transfer the solution to a 500 mL - resolution: minimum 1.4 between the peaks due to
separating funnel, rinsing the round-bottomed flask with ergocalciferol and cholecalciferol in the chromatogram
about 15 mL of a 10 gIL solution of sodium chloride Rand obtained with reference solution (b);
then with 150 mL of a mixture of equal volumes of ether R - the results obtained with the test solution (b) duplicates
and light petroleum Rl. Shake for 1 min. When the layers do not differ by more than 5 per cent.
have separated completely, discard the lower layerand wash Calculate the content of vitamin D 3 in International Units
the upper layer, first with 50 mL of a 30 gIL solution of per gram using the following expression) taking into account
potassium hydroxide R in a 10 per cent VIV solution of the assigned content of cholecalciferol CRS:
anhydrous ethanol R, and then with 3 quantities, each of
50 mL, of a 10 gIL solution of sodium chloride R. Filter the A2 x A3 x m2 x V2 x40
- JX A2
A, A 4 - [A 5 VI
upper layer through 5 g of anhydrous sodium su/fa.. R on a ml
fast filter paper into a 250 mL flask. Wash the funnel with AI
10 mL of fresh extraction mixture) filter and combine the In. mass of the substance to be examined in test solution (b). in
upper layers. Distil them at a temperature not exceeding grams;
30 °C under reduced pressure and fill with nitrogen R when 1n2 total mass of cho1«aki/erol CRS used for lhe preparation of
reference solution (a). in micrograms (500 J1g};
evaporation is completed. Alternatively, evaporate the solvent
AI area (or height) of the peak due to cholecakiferol in the
under a gentle current of nitrogen R at a temperature not chromatogram cbrelned with test solution (I);
exceeding 30 °C. Dissolve the residue in J.5 mL of the A2 area (or height) of the peak due to cholecalciferol in the
mobile phase described under Purification. Gentle heating in chromatogram obtained with test solution (b)j
AJ area (or height) of the peak due to ergocalciferol in the
an ultrasonic bath may be required. A large fractio» of the
white residue is cholesterol.. constituting approximattly A4 area (or heigh!) of the peak due to ergocalciferol in the
50 per centmlm of 'he unsaponifiable matterof cod-/iver oil; chromatogram obtained with test solution (b);
Testsdution (b) Prepare duplicates. To 4.00 g of the A, area (or height) of a possible peak in the chromatogram
obtained wiih test solution (a) with the same retention lime as
substance to be examined add 2.0 mL of the internal the peak co-eluting with ergoca.lciferol in test solution (b)j
standard solution and proceed as described for test A6 area (or height) of the peak due to cholecakiferol in the
solution (a). chromatogram obtained with reference solution (b);
VI total volume of reference solution (a) (100 mL);
Reference sduuon (a) Dissolve 0.50 mg of cholecalciferol CRS V2 volume of reference solution (a) used to prepare reference
in anhydrous ethanol R and dilute to 100.0 mL with the same solution (b) (2.0 mL).
solvent.
Reference solution (b) Into a round-bottomed flask, add STORAGE
2.0 mL of reference solution (a) and 2.0 mL of the intemal In an airtight and well-filled container) protected from light.
standard solution and proceed as described for test H no antioxidant is added, store under an inert gas,
solution (a). Once the container has been opened) its contents are used as
PURIFICATION soon as possible and any part of the contents not used at
Column: once is protected by an atmosphere of inert gas.
- size: I ;;;; 0,25 m, 0 = 4.6 mrn;
LABELLING
- stationary phase: nitrile silica gelfor chromatography R
The label states:
(10 pm).
- the number of International Units of vitamin A per gram;
Mobile phase isoamyl akohol R, hexaneR (1.6:98.4 VII'). - the number of International Units of vitamin D 3 per
FWw rate 1.1 mllmin. gram.
Detection Spectrophotometer at 265 om. ________ ~ PhE..
Injection 350 ~L of reference solution (b) and test

until 2 min after the retention time of cholecalciferol) in a
ground-glass-stoppered tube containing 1 mL of a J gIL
solution of butylhydroxyteluene R in hexaneR. Evaporate
separately to dryness at a temperature not exceeding 30 °C
under a gentle current of nitrogen R. Dissolve each residue in
1.5 mL of acetonitrile R.
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1-668 Cod-liver Oil (Type B) 2022
Use starch solution R2.

Cod-liver Oil (Type B)
Peroxide value (2.S.5. Method B)
(Ph. Eur. monograph 1193) Maximum 10.0.
Action and use Unsaponifiable matter (2.5.7)
Source of vitamins A and D. Maximum 1.5 per cent. determined on 5.0 g.
Each ill of vitamin D] is equivalent to 0.025 I!g of Stearin
colecalciferol. Heat at least 10 mL to 60-90 °C and cool to room
temperature. Immerse for 3 h in a bath of iced water or a
Ph€1r _
thermostatically controUed bath at 0 ± 0.5 "C. If necessary.
DEFINITION to eliminate insoluble matter. filter the sample after heating.
Purified fatty oil obtained from the fresh livers of wild cod. The sample remains clear.
Gadus morhua L. and other species of Gadidae. solid Composition of fatty acids
substances being removed by cooling and filtering. A suitable Gas chromatography (2.2.28).
antioxidant may be added.
Content Trl\'i.aI name of Nomenelatun: Lewer limlt Upper 1lmIt
fatty acid area area (per cent)
- vitamin A: 600 ill (l80 1Jg) to 2500 ill (750 I!g) per (per cent)
gram;
SalurauJ jatlY acitb:
- tHtamin D~: 60 ill (1.5 ug) to 250 ill (6.25 ug) per gram.
Myristic acid 14:0 2.0 6.0
PRODUCTION Palmitic add 16:0 7.0 .... 0
Stearic acid 18:0 1.0 4.0
The content of dioxins and dioxin-like PCBs
Mono-unsalldduJjally adds:
(polychlorinated biphenyls) is controUed using methods and
Palmltoleic acid 16:1 n-7 4.5 11.5
limits in accordance with the requirements set in the
as- Varo:nic acid 18:1 0-7 2.0 7.0
European Union or other applicable regulations. Oleic acid 18:10-9 12.0 21.0
Gadoleic acid 20:1 noll 1.0 5.5
CHARACTERS
Goodoic acid 20:10-9 5.0 17.0
Appearance Erucic acid 22:10-9 0 1.5
Clear. yellowish liquid. Celoleic acid (22: I 22:1 n-II+l3 5.0 12.0
Solubility n-I l)
PoI~unsal'lTtl,1Jd jalty acids:
Practically insoluble in water. miscible with light petroleum.
slightly soluble in ethanol ~96 per cent). linoleic acid 18:20-6 0.5 3.0
a-linolenic acid 18:30-3 0 2.0
IDENTIFICATION Moroctic acid 18:40-3 0.5 4.5
First identificatWn: A, B. C. Timnodonjc 20:5 D·3 7.0 16.0
(ejcosapenraenoie)
Second identi./icalion: C. D. acid (EPA)
A. In the assay for vitamin A using method A, the test Cervonic 22:6 n-3 6.0 18.0
solution shows an absorption maximum (2.2.25) at (docosahexaenoic)
325 ± 2 run. In the assay for vitamin A using method B. the acid (OHA)
chromatogram obtained with the test solution shows a peak

corresponding to the peak due to aU-trans-retinol in the Test solution Dilute about 0.45 g of the substance to be
chromatogram obtained with the reference solution. examined to 10.0 mL with a 50 mgIL solution of
B. In the assay for vitamin D 3 • the chromatogram obtained butylhydroxywluene R in nimethylpentane R. Transfer 2.0 mL
with test solution (a) shows a peak corresponding to the peak of this solution to a quartz tube and evaporate the solvent
due to cholecalciferol in the chromatogram obtained with with a gentle current of nitrogen R. Add 1.5 mL of a 20 gIL
reference solution (b). solution of sodium hydroxide R in methanol R, cover with
nitrogen R, cap tightly with a polytetrafluoroethylene-lined
cap, mix and heat on a water-bath for 7 min. Cool, add
D. To 0.1 g add 0.5 mL of methy/emchloride Rand 1 mL of 2 mL of boron trichloride-methanol solution R. cover with
antimonytn'chloride solution R. Mix. A deep blue colour nitrogen R. cap tightly, mix and heat on a water-bath for
develops in about lOs. 30 min. Cool to 40-50 °C, add 1 mL of trimethylpenUine R,
TESTS cap and vortex: or shake vigorously for at least 30 s.
Appearance Immediately add 5 mL of saturated sodium chloride solution R.
The substance to be examined is not more intensely coloured cover with nitrogen R. cap and vortex or shake thoroughly for
than a reference solution prepared as follows: to 3.0 mL of at least 15 s. Anow the upper layer to become clear and
red primary solution add 25.0 mL of yellow primary solution transfer to a separate tube. Shake the methanol layer once
and dilute to 50.0 mL with a 10 gIL solution of hydrochlmic more with 1 mL of m'methy/pentane R and combine the
acidR (2.2.2. Method 11). rrimethylpentane extracts. Wash the combined extracts with
2 quantities, each of 1 mL, of water R and dry over anhydrous
sodium sulfate R. Prepare 2 solutions for each sample.
0.917 to 0.930.
Reference solution Dissolve 0.300 g of methylarachidase R.
0.300 g of merhyl behenate R. 0.300 g of methylpalmitate R
1.477 to 1.484.
and 0.300 g of methyl stearate R in a 50 mgIL solution of
Acid value (2.5.1) butylhydroxywluene R in m'memylpenUine Rand dllute to
Maximum 2.0. determined on 20.00 g. 10.0 mL with the same solution.
Iodine value (2.5.4. jHethod B) Column:
150 to 180. determined on 0.14 g to 0.16 g. - material: fused silica;
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2022 Cod-liver Oil (Type B) 1-669
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~ n • 0
t;o
N "u
N ,;
~ 7i~ri
n U
m ~ N U
~
I , , u N
I ~
u
• •
~
I
, I I I I
10 20 30 40 50 60 min
Figure 1193.-1. - Chromatogram for the test for composition of fatty acids of cod-liver oil (type B)
- size: / = 30 m, 0 = 0.25 mm; -

minimum 1.3 between the peaks due to methyl oleate
- stationary phase: mcu.rogol 20 000 R (film thickness and methylru-vaccenatei the resolution between the
0.25 urn). peaks due to methyl gadoleare and methyl gondoate is
Carrier gas hydrogen for chromatography R or helium for sufficient for purposes of identification and area
chromalOgraphy R, with an oxygen scrubber. measurement.
Split ratio 1:200. Calculate the area per cent for each fatty acid methylester
using the following expression:
Temperature:
Time
(min)
Temperature
rq
~: x 100
Column 170 -i 225
A~ peakareaof falty acid X;
225 A, sum of me peak areas (up to C22:6 n-3).
Injection port 250
Detector 280 The calculation is not valid unless:
- the total area is based only on peaks due solely to fatty
Dueaion Flame ionisation. acid methyl esters;
- the number of fatty acid methylesterpeaks exceeding
Injection 1 ~L, twice.
0.05 per cent of the total area is at least 24;
System suitabil,;y: - the 24 largest peaks of the methyl esters account for more
- the 15 fatty acids to he tested are satisfactorily identified than 90 per cent of the total area (thesecorrespond to, in
from the chromatogram shown in Figure 1193.:-1 j common elution order: 14:0, 15:0, 16:0, 16:1 0-7,
- in the chromatogram obtained with the reference solution, 16:4 n-I, 18:0, 18:1 n-9, 18:1 n-7, 18:2 n-6, 18:3 n-S,
multiply the areas of the peaks due to methyl palmitate, 18:4 n-3, 20:1 n-11, 20:1 n-9, 20:1 n-7, 20:2 n-e, 20:4
methyl stearate, methyl arachidate and methyl behenate n-6, 20:3 n-3, 20:4 n-3, 20:5 n-3, 22:10-11,22:1 n-9,
by the corresponding response factors in Table 1193.-1; 21:5 n-S, 22:5 n-3, 22:6 n-3).
normalise the corrected areas of the peaks due to the fatty
acid methyl esters to a sum of 100 per cent; the ASSAY
normalised area percentage of each fatty acid methylester Vitamin A
is to be within ± 1.0 percentage unit of the Cany out the test as rapidly as possible, awiding exposure to
corresponding weight percentage; actinic light and air, oxidising agents.. oxidation catalysts (for
example, <»J>Per and iron) and acids.
Table 1193.-1. Use method A. If method A is found not to be valid, use
Fatty acid methyl ester Theoretical response factor method B.
Melbyl palmitate 1.049 METHOD A
Methyl stearate 1.029 Ultraviolet absorption spectrophotometry (2.2.25).
Melbyl arachidate 1.0B
Test,oIut.,n To 1.00 g in a round-bottomed flask, add
Melbyl behenete l.000
3 mL of a freshly prepared 50 per cent mlm solution of
potassium hydroxide Rand 30 mL of anhydrous ethanol R. Boil
- resolution: in the chromatogram obtained with the test under reflux in a current of nitrogen R for 30 min. Cool
solution: rapidly and add 30 mL of wat<r R. Extract with 50 mL of
ether R. Repeat the extraction 3 times and discard the lower
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1-670 Cod-liver Oil (Type B) 2022
layer after complete separation. Wash the combined upper and fill with nitrogen R when evaporation is completed.
layers with 4 quantities, each of 50 mL, of water Rand Alternatively, evaporate the solvent under a gentle current of
evaporate to dryness under a gentle current of nitrogen R at a nitrogen R at a temperature not exceeding 30 °C. Dissolve the
temperature not exceeding 30°C or by other suitable means residue in 2-propanol Rl, transfer to a 25 mL volwnetric flask
at a temperature not exceeding 30°C under reduced and dilute to 25 mL with 2-propanol RI. Gentle heating in an
pressure. Dissolve the residue in sufficient 2-propanol Rl to ultrasonic bath may be required. A large fraction of the white
give an expected concentration of vitamin A equivalent to residue is cholesterol, constiuuing approximately 50 per cent mlm
10-15 IU/mL. of the umaponifiable mauer of cod-liver ail.
Measure the absorbances of the solution at 300 run, 310 run, .Reference solution (a) Prepare a solution of retinol
325 run and 334 run and at the wavelength of maximum acetate CRS in 2-propano! Rl so that 1 mL contains about
absorption with a suitable spectrophotometer in specially 1000 IU of atl-tram-retinol.
matched 1 em cells, using 2-propano! Rl as the compensation The exact concentration of reference solution (a) is assessed
liquid. by ultraviolet absorption spectrophotometry (2.2.25). Dilute
Calculate the content of vitamin A, as all-trans-retinol, in reference solution (a) with 2-propano! RJ to a presumed
International Units per gram using the following expression: concentration of 10-15 IU/mL and measure the absorbance
at 326 om in matched 1 cm ceUs using 2-propano/ Rt as the
1821
A 325 X - - xV compensation liquid.
100m
Calculate the content of vitamin A in International Units per
Al H absorbance at 325 nm, millilitre of reference solution (a) using the following
m to
mass of the substance be examined. in grams; expression, taking into account the assigned content of retinol
V total volume of solution containing 10-15 ill ofvitunin A per
millilitre;
aatate CRS:
1821 conversion factor foe the specific absorbance of aU-tmns-retinol,
in International Units. A 1900xV,
326 x 100x VI
The above expression can be used only if A 325 has a value II,,. absorbance at 326 run;
not greater than A 32 5•con!O.970 where A 32 5,corr is the V, volume of reference solution (a) used;
corrected absorbance at 325 run and is given by the following V, volume of the dihned solution;
equation: 1900 conversion factor foe the specific absorbance of retinol
autatt CRS. in International Ullin.
A"'.<QIT = 6.815A'25 - 2.555A3Io - 4.260A'34
A designates the absorbance at the wavelength indicated by solution but using 2.00 mL of reference solution (a) in place
the subscript. of the substance to be examined.
If A 32 5 has a value greater than A.325,con!0.970, calculate the The exact concentration of reference solution (b) is assessed
content of vitamin A using the foUowing expression: by ultraviolet absorption spectrophotometry (2.2.25). Dilute
reference solution (b) with 2-propanol RI to a presumed
1821
A.325,coIT x 100m x V concentration of 10-15 IU/mL of aU-trans-retinol and
measure the absorbance at 325 om in matched 1 em cells
The assay is not valid unless: using 2-propanol RJ as the compensation liquid.
- the wavelength of maximum absorption lies between Calculate the content of aU-trans-retinol in International
323 run and 327 nm; Units per millilitre of reference solution (b) using the
- the absorbance at 300 run relative to that at 325 run is at following expression:
most 0.73.
A 1821 X V,
METHODB 325 X 100x V4
AJ 2S absorbance at 325 nm;
Test solution Prepare duplicates. To 2.00 g of the substance VJ volume of the diluted solution;
to be examined in a round-bottomed flask, add 5 mL of a Vol volume of reference solution (b) used;
fresWy prepared 100 gIL solution of ascm-bic acid R, 10 mL of 1821 conversion factor for the specific absorbance ofaU-mms-retinol.
a freshly prepared 800 gIL solution of potassium hydroxide R in International Units.
and 100 mL of anhydrous ethanol R. Boil under a reflux
condenser on a water-bath for 15 min. Add 100 mL of a Column:
10 gIL solution.of sodium chloride R and cool. Transfer the - size: 1:::::: 0.25 m, 0 : : : 4.6 mm;
solution to a 500 mL separating funnel, rinsing the round- - suuionaty phase: octade<ylsilyl silica gelfor chromOlography R
bottomed flask with about 75 mL of a 10 gIL solution of (5-10 pm).
sodium chloride R and then with ISO mL of a mixture of Mobile phase warer for chromatography R, methanol R
equal volumes of ether R and lightperroleum RI. Shake for (3:97 VIV).
I min. When the layers have separated completely, discard FWtiJ rate I mLImin.
the lower layer and wash the upper layer, first with 50 mL of Detection Spectrophotometer at 325 run.
a 30 gIL solution of potassium hydroxUk R in a
10 per cent VIV solution of anhydrous ethanol R and then
Injection 10).11..; inject in triplicate the test solution and
with 3 quantities, each of 50 mL, of a 10 gIL solution of
sodium chloride R. Filter the upper layer through 5 g of Reuntion time AlI-trans-retinol:::::: 5 ± 1 min.
anhydrous sodium sulfate R on a fast filter paper into a Sysum suitability:
250 mL flask.Wash the funnel with 10 mL of fresh extraction - the chromatogram obtained with the test solution shows a
mixture, filter and combine the upper layers. Distil them at a peak corresponding to the peak due to ell-neas-retinol in
temperature not exceeding 30 °C under reduced pressure the chromatogram obtained with reference solution (b);
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2022 Cod-liver Oil (Type B) 1-671
- the results obtained with the duplicate test solutions do PURIFICATION

not differ by more than 5 per cent; Column:
- the recovery of all-trans-retinol in reference solution (b) as - size: 1= 0.25 m, 0 = 4.6 rnm;
assessed by direct absorption spectrophotometry is greater - stationary phase: nirn'le silica gelfor chromatography R
than 95 per cent. (10 IJm).
Calculate the content of vitamin A using the following Mobile phase isoamyl akoho/ R, hexane R (1.6:98.4 VIV).
expression: Flow rale 1.1 mUmin.
c« V 1 Detection Spectrophotometer at 265 run.
Alx~-x-
Az m Injection 350 J.lL of reference solution (b) and test
A, area of the peak due to aU·tmns-~tinol in the chromatogram until 2 min after the retention time of cholecalciferol, in a
obtained with the test solutionj
A2 area of the peak due 10 aU-Iram-retinol in the chroma togram
ground-glass-stoppered tube containing 1 mL of a 1 gIL
obtained with reference solution (b); solution of butylhydroxytoluene R in hexane R. Evaporate
C concentration of reti1lol aceuue CRS in reference solution <a) as separately to dryness at a temperature not exceeding 30 °C
assessed prior to the sapooification, in International Units per under a gentle current of nitrogen R. Dissolve each residue in
rnilJilitre (= 1000 IUfmL);
1.5 mL of acetonitrile R.
V volume of reference solution (a) Treated (2.00 mL);
til mass of the substance to be examined in the test solution DETERMINATION
(2.00 g). Column:
- size: 1= 0.15 m, 0 4.6 mm;=
Vitamin D) - stationary phase: octadecylsilyl sitica gelfor chromatography R
Liquid chromatography (2.2.29). Carryout the assay as rapidly . (5 urn),
as possible, avoiding exposure to actinic light and air. Mobile phase phosphoric acidR, 96 per cent VIV solution of
Internal standard solution Dissolve 0.50 mg of aatonitri/eR (0.2:99.8 VIV).
ergocalciferol CRS in anhydrous ethanol R and dilute to Flaw rate 1.0 mUmin.
Testsolwion (a) To 4.00 g of the substance to be examined
in a round-bottomed flask, add 5 mL of a freshly prepared Injection 2 quantities not exceeding 200 I!L of each of the
100 gIL solution of ascorbic acid R, 10 mL of a freshly 3 solutions obtained under Purification.
prepared 800 gil.. solution of potassium hydroxide Rand System suitability:
100 mL of anhydrous ethanolR. Boil under a reflux - resolution: minimum 1.4 between the peaks due to
condenser on a water-bath for 30 min. Add 100 mL of a ergocalciferol and cholecalciferol in the chromatogram
10 gIL solution of sodium chloride R and cool the solution to obtained with reference solution (b);
room temperature. Transfer the solution to a 500 mL - the results obtained with the test solution (b) duplicates
separating funnel, rinsing the round-bottomed flask with do not differ by more than 5 per cent.
about 75 mL of a to gIL. solution of sodium chloride Rand Calculate the content of vitamin D 3 in International Units
then with 150 mL of a mixture of equal volumes of ether R per gram using the following expression, taking into account
and Tight petroleum R 1. Shake for 1 min. When the layers the assigned content of cholecalciferol CRS:
have separated completely, discard the lower layer and wash
the upper layer, first with 50 mL of a 30 gil.. solution of A2 x A) x m2 x V2 x 40
potassium hydroxide R in a 10 per cent VIV solution of A6 A4 _ [~~] xA2 ml VI
anhydrous ethanol R. and then with 3 quantities, each of
50 ml., of a 10 gil.. solution of sodium chloride R. Filter rue
upper layer through 5 g of anhydrous sodium sulfate R on a mI mass of the substance to be examined in tes! solution (b), in
grams;
fast filter paper into a 250 mL flask.Wash the funnel with
m, lolal mass of droleaJ1cijerol CRS used for the preparation of
10 mL of fresh extraction mixture, filler and combine the reference solution (8), in micrograms (500 "g);
upper layers. Distil them at a temperature not exceeding A, area (or height) of the peak due 10 cholecalciferol in the
30 °C under reduced pressure and fill with nitrogen R when chromatogram obtained with test solution (a);
evaporation is completed. Alternatively, evaporate the solvent A, area (or height) of the peak due to cholecalciferol in the
chromatogram obtained with lest solution (b);
under a gentle current of nitrogen R at a temperature not A} = area (or height) of the peak due EO ergocalciferol in the
exceeding 30 °C. Dissolve the residue in 1.5 mL of the chromatogram obtained with reference solunon (b);
mobile phase described under Purification. Gentle heating in A4 area (or height) of the peak due to ergocakilerol in the
an ultrasonic bath may be required. A Targe fraction of the chromatogram obtained with lest solution (b);
A; area (or height) of a possible peak in the chromatogram
white residue is cholesterol, constituting approximately obtained with lest solulion (a) with the same retention time as
50 per cent mfm of the unsaponifiable matur of cod-liver oil. the peak co-eluting with ergocalciferol in lest solution (b)j
Testsolution (b) Prepare duplicates. To 4.00 g of rue A6 area (or height) of the peak due to cholecalciferol in the
chromatogram obtained with reference solution (bl;
substance to be examined add 2.0 mL of the internal VI total volume of reference solution (a) (100 rnL);
standard solution and proceed as described for test V, volume of reference solution (a) used to prepare reference
solution (a). solution (b) (2.0 rnL),
Reference solution (a) Dissolve 0.50 mg of choTeca/aleroi CRS
in anhydrous ethanol R and dilute to 100.0 mL with the same STORAGE
solvent. In an airtight and well-filled container, protected from light.
Reference solution (b) In a round-bottomed flask, add If no antioxidant is added, store under an inert gas.
2.0 mL of reference solution (a) and 2.0 mL of the internal Once the container has been opened, its contents are used as
standard solution and proceed as described for test soon as possible and any part of the contents not used at
solution (a). once is protected by an atmosphere of inert gas.
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1-672 Colchicine 2022
LABELLING TESTS
The label states: Soludon S
- me number of International Units of vitamin A per gram; Dissolve 0.10 g in carbon dioxide-free water R and dilute to
- the number of International Units of vitamin D J per 20 mL with the Same solvent.
gram. Appearance of solution
_ _ _~ ~~~~~ PfJEI¥ Solution S is clear (2.2.1) and not more intensely coloured
than reference solution GY 3 (2.2.2, Method II).
Colchicine solution R J. Either the solution does not change colour or it
becomes green. Not more than 0.1 mL of 0.01 M sodium
(Ph. Eur. monograph 0758) hydroxide is required to change the colour of the indicator to
blue.
H3CO~H
••N
y CH ] Specific optical rotation (2.2.7)
...", H -250 to -235 (anhydrous substance).
H]CO ~ a
H3CO Dissolve 50.0 mg in ethanol (96 per cent) R and dilute to
~ a 10.0 mL with the same solvent.
OCH 3 Related substances
399.4 64-86-8 Solvent mixture methanol R, water R (50:50 VIP).
Testsdution Dissolve 20.0 mg of the substance to be
Action and use examined in the solvent mixture and dilute to 20.0 mL with
Used in treatment of gout. the solvent mixture.
Preparation Reference solution (a) Dissolve 5 mg of cokhicine for system
Colchicine Tablets suitability A CRS (containing impurities A, E and G) in the
solvent mixture and dilute to 5.0 mL with the solvent
PhEtr _ _ ~~ ~ ~_
mixture.
DEFINITION Reference solution (b) Dilute 1.0 mL of the test solution to
N-l(7S,12aM)-1,2,3,1 0-tetramethoxy-9-oxo-5,6,7,9- 100.0 mL with the solvent mixture.
tetrahydrobenzo[alheptalen-7-yll acetamide. Column:
Content - size: 1= 0.25 m, 0 =4.6 mm;
97.0 per cent to 102.0 per cent (anhydrous substance). - stationary phase: octy/sl"lyl silica gel for chromatography RJ
(5 J.lm)j
CHARACTERS
Appearance
Yellowish-white, amorphous or crystalline powder. Mobile phase Mix 450 volumes ofa 6.8 gIL solution of
potassium dihydrogen phosphate R and 530 volumes of
Solubility methanol R. After cooling to room temperature, adjust the
Very soluble in water, rapidly recrystaUising from volume to 1000 mL with methanol R. Adjust the apparent pH
concentrated solutions as me sesquihydrate, freely soluble in to 5.5 with dilute Phosphoric acid R.
ethanol (96 per cent), practically insoluble in cyclohexane.
Flow rate 1 mUmin.
IDENTIFICATION Detection Spectrophotometer at 254 run.
Injution 20 J.!L.
Run time 3 times the retention time of colchicine.
Identificau01l of impurities Use the chromatogram supplied
(2.2.25).
with colchicine for system suitability A CRS and the
Test solutwn Dissolve 5 mg in ethanol (96 per cent) R and chromatogram obtained with reference solution (a) to
dilute to 100.0 mL with the same solvent. Dilute 5.0 mL of identify the peaks due to impurities A, B, E and G.
this solution to 25.0 mL with ethanol (96 percent) R. (Impurity B is the conformational isomer of colchicine, which
Spectral range 230-400 om. is formed insitu in solution).
Absorption maxima 243 om and 350 run. Relative retention With reference to colchicine (retention
Absorbance ratio A 24JA350 = 1.7 to 1.9. time = about 7 min): impurity E = about 0.6;
B. Infrared absorption spectrophotometry (2.2.24). = =
impurity B about 0.9; impurity A about 0.94j
Comparison colchicine CRS.
=
impurity G about 1.4.
C. To 0.5 mL of solution S (see Tests) add 0.5 mL of dilute =
- peak-IO-valley ratio: minimum 2, where Hp height above
hydrochloric acid Rand 0.15 mL of ferric chloride solution RI. the baseline of the peak due to impurity A and
The solution is yelIow and becomes dark green on boiling for
30 s. Cool, add 2 mL of methylene chloride R and shake.
=
H" height above the baseline of the lowest point of the
The organic layer is greenish-yelIow.
colchicine; minimum 2, where Hp = height above the
D. Dissolve about 30 mg in I mL of ethanol (96 per cent) R =
baseline of the peak due to impurity Band H" height
and add 0.15 mL offeme chloride solution RI. A brownish-red above me baseline of the lowest point of the curve
colour develops. separating this peak from the peak due to impurity A.
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2022 Colchicine 1-673

- correction factor: multiply the peak area of impurity G by
1.6;
- for each impurity, use the concenrration of colchicine in
reference solution (b). o
Limits: OCH3
- impurjry A: maximum 3.0 per cent;
B. N-[(7S, 12aP)-l,2,3,1 0-tetramethoxy-9-oxo-5,6,7,9-
- impurity G: maximum 0.25 per cent;
tetrahydrobenzo(a]heptalen-7-yl]acetamide
- impurity E: maximum 0.2 per cent;
(conformational isomer),
0.10 per cent; HaCO
- reponing threshold: 0.05 per cent; disregard the peak due to
impurity B.
Impurity F
o
Maximum 0.2 per cent. OCHa
Dissolve 50 mg in water R and dilute to 5 mL with the same
solvent. Add 0.1 mL offmie chloride sdution Rl, C. N- [(7S, 7bR, lOaS)-l ,2,3, 9-tetramethoxy-8-oxo-
The solution is not more intensely coloured than a mixture 5,6,7,7b,8,1 Oa-hexahydrobenzo[a]cycl openta[3,4]
of I mL of red primary solution, 2 mL of yellow primary cyclobuta[I,2-c]cyclohepten-7-yl]acetamide
solution and 2 mL of blue primary solution (2.2.2, (P-Iumicolchicine),
:~:
Method If).
Ethyl acetate (2.4.24)
Maximum 6.0 per cent m/m.
Water (2.5.12)
Maximum 2.0 per cent, determined on 0.500 g. OH H3 CO
Sulfated ash (2.4.14) o
Maximum 0.1 per cent, determined on 0.5 g. OCH 3
ASSAY D. N-[(7 S, 12aM)-3-(p-n-gjucopyranosyloxy)-1 ,2,1 0-
Dissolve 0.250 g with gentle heating in a mixture of 20 mL trimethoxy-9-oxo-5,6,7,9-terrahydrobenzo[a]heptalen-7-yl]
of acetic anhydride R and 40 mL of toluene R. Titrate with acetamide (cokhicoside),
0.1 M perchloric add, determining the end-point
HO
1 mL of 0.1 M perch/orit add is equivalent to 39.94 mg of
C22H2~06'
STORAGE
OCH3
°
IMPURITIES
E. N- (7S, 12aM)-3-hydroxy-I,2, I 0-trimethoxy-9-oxo-
Spedfied impurities A, E, F, G.
5,6,7,9-tetrahydrobenzo[a]heptalen-7-yl]acetamide (3-0-
Other detectable impurities (the following substonces would, if demethylcolchicine),
present at a sufficient level, be detected by one or otherof the tests
C
H3 °5Q:
1 .':yCH
in the monograph. They all!limited by the general acceptance l
monograph Substances for phannaceutical use (2034). It is HaCO ,0
thell!fore not necessary to identify these impurities for HaCO
demonstration of compliance. See also 5.10. Control of impurities =- 0

in substances forpharmaceutical use) B, C, D. OH
F. N-[(7S,12aM)-1 D-hydroxy-l,2,3-trimethoxy-9-oxo-
5,6,7,9-tetrahydrobenzo[a]heptalen-7-yl]acetamide
(colchiceine),
H3CO
o
A. N- [(7S, 12aM)-I,2,3,lO-tetramethoxy-9-oxo-5,6,7,9-
tetrahydrobenzo[a]hep13len-7-yl]formamide (N-deacetyl- OCH3
N-formyfcolcb.icine),
G. N-[ (7S, 7bS, IOaR)-1,2,3,9-tetramethoxy-8-oxo-
5,6,7,7 b,8, 10a-hexahydrobenzo(a]cyclopenta[3,4]
cyclobuta[1,2-c]cydohepten-7-yl]acetamide
(y-Iumicolchicine).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE/J{
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1-674 Colecalciferol 2022
corecarciferol ****••
*
Related substances
(Chol«a1ciftrol, Ph. Eur. monograph 0072)
**.** immediately before use, avoiding exposure to actinic light and air.
examined in trimethylpentane R without heating and dilute [0
Reference solution (a) Dissolve 10.0 mg of cholecalciferol CRS
in trimethylpentane R without heating and dilute to 10.0 mL
Reference solution (b) Dilute 1.0 mL of cholaalciferol for
syuem suitability CRS (containing impurity A) to 5.0 mL with
the mobile phase. Heat in a water-bath at 90°C under a
reflux condenser for 45 min and cool (formation of pre-
cholecalciferol).
Reference solution (c) Dilute 10.0 mL of reference
384.6 67-97-0 solution (a) to 100.0 mL with the mobile phase. Dilute
1.0 mL of this solution to 100.0 mL with me mobile phase.
Action and use
Column:
Vitamin D3 analogue.
- size: 1= 0.25 rn, 0 = 4.6 rom;
Preparations - stationary phase: silka gelfor chromatography R (5 J.lI1l).
AIendronic Acid and Colecalciferol Tablets
Mobile phase pentanol R, hexaneR (0.3:99.7 VIV).
Calcium and Colecalciferol Tablets
Flow rate 2 mUmin.
Calcium and Colecalciferol Chewable Tablets
Detection Spectrophotometer at 265 nm,
Colecalciferol Injection
Colecalciferol Tablets solutions (b) and (c).
Paediatric Vitamins A, C and D Oral Drops Run time Twice the retention time of cholecalciferol.
When cholecalciferol or vitamin D3 is prescribed or Relative retention With reference to cholecalciferol (retention
demanded, Colecalciferol shall be dispensed or supplied.
When calciferol or vitamin D is prescribed or demanded,
= =
time about 19 min): pre-cholecalciferol about 0.5;
Colecalciferol or Ergocalciferol shall be dispensed or
supplied.
- resolution: minimum 1.5 between the peaks due to pre-
PlJEu _ cholecalciferol and impurity A.
Limits:
DEFINITION
- impuriIJI A: not more than the area of the principal peak
(5Z,7E)-9, I o-Secocholesta-5,7,1 0(l9)-trien-3Jl-ol.
Content (0.1 per cent);
97.0 per cent to 102.0 per cent. - unspedfied impurities: for each impurity, not more than the
A reversible isomerisation to pre-cholecalciferol takes place in area of the principal peak in the chromatogram obtained
solution, depending on temperature and time. The activity is with reference solution (c) (0.10 per cent);
due to both compounds (see Assay). - total: not more than 10 times the area of the principal
I mg of cholecalciferol is equivalent to 40 000 IU of peak in the chromatogram obtained with reference
antirachitic activity (vitamin D) in rats. solution (c) (1.0 per cent);
CHARACTERS the chromatogram obtained with reference solution (c)
Appearance (0.05 per cent); disregard the peak due to pre-
White or almost white crystals. cholecalciferol.
Solubility
ASSAY
(96 per cent), soluble in trimethylpentane and in fatty oils.
It is sensitive to air, heat and light. Solutions in solvents
Injeaion Test solution and reference solution (a).
without an antioxidant are unstable and are to be used
immediately. Calculate the percentage content of G.l1H440 taking into
account the assigned content of chole«Jlciferol CRS and, if
IDENTIFICATION necessary, the peak due to pre-cholecalciferol.
STORAGE
Comparison cholcrolciferol CRS.
Under nitrogen, in an airtight container, protected from light,
TESTS at a temperature of 2 DC to 8°C.
Specific optical rotation (2.2.7) The contents of an opened container are to be used
+ 105 to + 112, determined within 30 min of preparing the immediately.
solution.
IMPURITIES
Dissolve 0.200 g rapidly in aldehyde-free alcohol R without
Specified ;mpun'ties A.
heating and dilute to 25.0 mL with the same solvent.
Otherdetectable impun'ties (th« following substances umdd, if
present at a rufficiem level, be detected by ane or other vf the tests
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2022 Colecalciferol 1-675

criterion for otherJunsp&::lfied impurities and/or by the general
Colecalciferol Concentrate (Oily
monograph Substances for pharmaceutical use (2034). It is Form)
therefore not necessary to identify these impun'ties for
(Cholecalciferol Concentrate (Oily Form), Ph, Eur.
monograph 0575)
in substances for pharmaceutical use) B, C, D, E.
Action and use
H)C-, H
Vitamin D analogue (Vitamin D).
CH3
Phfll _
DEFINITION
Solution of ChokcaJcijerol (0072) in a suitable vegetable fatty
oil, authorised by the competent authority.
Content
90.0 per cent to 110.0 per cent of the cholecalciferol content
" OH
H stated on the label, which is not less than 500 000 IU/g.
It may contain suitable stabilisers such as antioxidants.
A. (5E,7 E) -9, I0-secocholesta-5,7,1 0(l9)-trien-3p-ol (trans-
cholecalciferol, trans-vitamin D 3 ) , CHARACTERS
Appearance
Clear, yellow liquid.
Solubility
Practically insoluble in water, slightly soluble in anhydrous
ethanol, miscible with solvents of fats,
HO :
Partial solidification may occur, depending on the
H temperature.
IDENTIFICATION
B. cholesta-5,7-dien-3 /l-ol (7,8-didehydrocholesterol,
provitamin D 3 ) , A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Test solution Prepare a solution in cycJohexane R containing
the equivalent of about 400 IU/mL.
Speetral range 250-300 run.
Absorption maximum At 267 run.
HO : B. Examine the chromatograms obtained in the assay.
H Results The principal peak in the chromatogram obtained
with the test solution is similar in retention time to the
C. 9~, I 0a-cholesta-5,7 -dien-Bjl-ol (lumisrerok),
principal peak in the chromatogram obtained with reference
solution (a).
TESTS
Acid value (2.5.1)
Maximum 2.0.
Dissolve 5.0 g in 25 mL of the prescribed mixture of
solvents.
Peroxide value (2.5.5, Method A)
Maximum 20.
Related substances
D. (6E)-9,1 O-secocholesta-5(1 0),6,8(1 4)-rrien-3p-ol (iso- (Table 2034.-1) in the general monograph Substances for
tachysterok), pharmaceutical use (2034) do not apply.
ASSAY
Cany out the assayas rapidly as possible, awiding exposure to
actinic light and air.
Test solution Dissolve a quantity of the preparation to be
examined, weighed with an accuracy of 0.1 per cent,
equivalent to about 400000 ill, in 10.0 mL of toluene Rand
Reference solution (a) Dissolve 10.0 mg of ehokrolciferol CRS
without heating in 10.0 rnL of toluene R and dilute to
E. (6E)-9,1 0-secocholesta-5(10),6,8-trien-31>-01 100.0 rnL with the mobile phase.
(tachysterolj). Reference solution (b) Dilute 1.0 mL of eholecakiferolfor
___________ ~ PhEw system suitability CRS to 5.0 mL with the mobile phase. Heat
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1-676 Colecalciferol 2022
in a water-bath at 90°C undera reflux condenser for 45 min STORAGE

and cool. In an airtight, well-filled container, protected from light.
Reference sohnion (c) Dissolve 0.10 g of cholecalciferol CRS The contents of an opened container areto be used as soon
without heating in toluene R and dilute to 100.0 mL with the as possible; any unused part is to be protected by an
same solvent. atmosphere of nitrogen.
Reference solution (d) Dilute 5.0 mL of reference solution (c) LABELLING
to 50.0 mL with the mobile phase. Keep the solution in iced The label states:
water. - the number of International Units per gram;
Reference solution (e) Place 5.0 mL of reference solution (c) - the method of restoring the solution if partial solidification
in a volumetric flask, add about 10 mg of occurs.
butylhydroxywlutne R and displace air from the flask with __ ~ PhE",
nilrOgen R. Heat in a water-bath at 90°C undera reflux
condenser protected from light and under nitrogen R for
45 min. Cool and dilute to 50.0 mL with the mobile phase.
Column:
- size: 1 = 0.25 rn, 0 = 4.6 rom;
Colecalciferol Concentrate ......"""......
- stationary phase: s«ka gelfor chromawgraphy R (5 urn). (Powder Form) .... "
Moinle phase pentanol R, hexaneR (3:997 VIV). (Cholecakllerol Concentrate (powder Form), Ph. Bur.
Flow rate 2 mUmin. monograph 0574)
Detection Spectrophotometer at 254 nm, Action and use
Injection The chosen volume of each solution(the same Vitamin D analogue (Vitamin D3) .
volume for reference solution (a) and for the test solution); PhE" _
automatic injection device or sample loop recommended.
Relative retention With reference to cholecalciferol (retention DEFINITION
time = about 19 min): pre-cholecalciferol := about0.4; trans- Powder concentrate obtained by dispersing an oily solution of
cholecalciferol := about 0.5. Cholecalcllerol (0072) in an appropriate matrix, which is
usually based on a combination of gelatin and carbohydrates
System suiwb«ity Reference solution (b):
of suitable quality, authorised by the competent authority.
- resolution: minimum 1.0 betweenthe peaks due to pre-
cholecalciferol and lJ'ans-cholecaJciferolj if necessary adjust Content
the proportions of the constituents and the flow rate of 90.0 per cent to 110.0 per cent of the cholecalciferol content
the mobilephase to obtain this resolution; stated on the label, which is not less than 100000 IV/g.
- repeatability: maximum relative standard deviation of It may contain suitable stabilisers such as antioxidants.
1.0 per cent for the peak due to cholecalciferol after CHARACTERS
6 injections.
Appearance
Calculate the conversion factor (j) usingthe following White or yellowish-white, small particles.
expression:
Solubility
K-L Practically insoluble, swells, or forms a dispersion in water,
M depending on the formulation.
K area(or height) of me peakdue to cholecalciferol in me

IDENTIFICATION
chromatogram obtained with reference solution(d); First idemification: A, C.
L area(or height) of the peak. due to cholecalciferol in the Second identification: A, B.
chromatogram obtained. with reference solution(e);
M area(or height) of the peakdue to pre-cholecalciferol in the A. Thin-layer chromatography (2.2.27). Prepare the solutions
chromatogram obtainedwith reference solution(e). immediately before use.
Test solution Place 10.0 mL of the testsolutionprepared for
The value off determined in duplicate on different days may the assay in a suitable flask and evaporate to dryness under
be used during the entire procedure. reduced pressure by swirling in a water-bath at 40°C. Cool
Calculate the content of cholecalciferol in International Units under running water and restore atmospheric pressure with
per gram using the following expression: nitrogen R. Dissolve the residue immediately in 0.4 mL of
ethylene chloride R containing 10 gIL of squalane Rand
m' x~x SD+(jXS,) x 40000 x 1000 0.1 gIL of butylhydroxyto/uene R.
V' m S"o Reference solution (a) Dissolve 10 mg of cholecalciferol CRS
in ethylene chloride R containing 10 gIL of squalane R and
m mass of the preparation to beexaminedin the test solution, in
milligrams; 0.1 gIL of butylhydroxyrolueue R and dilute to 4 mL with the
m' mass of choka1lri/trol CRS in reference solution (a), in same solution.
milligrams;
V volume of the test solution (100 mL);
Reference solutio.. (b) Dissolve 10 mg of ergocakjJeroi CRS in
Vi volume of reference solution (a) (100 mL); ethylene chloride R containing 10 gIL of squalane Rand·
SD area(or height) of me peak due LO cholecalciferol in the 0.1 gIL of butylhydroxywluene R and dilute to 4 mL with the
chromatogram obtained with the test solution; same solution.
S'D area. (or height) of the peakdue to cholecalciferol in the
chromatogram obtainedwith reference solution(a); Plate TLC silica gel G plateR.
Sp area (or height) of the peak. due to pre-cholecalciferol in the Mob«ephase A 0.1 gIL solution of butylhydroxywluene R in a
chromatogram obtainedwith the test solution; mixture of equal volumes of cyciohexane R and peroxide-free
f conversion factor.
ether R.
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2022 Colecalciferol 1-677
Application 20 ~L. the washings have an approximate pH of 7 to 8) using a pH

Development Immediately, protected from light, over a path indicator strip R. Transfer the washed pentane extract to a
of 15 em. ground-glass-stoppered flask. Evaporate the contents of the
flask to dryness under reduced pressure by swirling in a
Drying In air.
water-bath at 40°C. Cool under running water and restore
Detection Spray with sulfuric acid R. atmospheric pressure with nurogen R. Dissolve the residue
Results The chromatogram obtained with the test solution immediately in 5.0 mL of toluene R and add 20.0 mL of the
shows immediately a bright yellow principal spot, which mobile phase [0 obtain a solution containing about
rapidly becomes orange-brown, then gradually greenish-grey, 4000 IU/mL.
remaining so for 10 min. This spot is similar in position, Reference solution (a) Dissolve 10.0 mg of cholecalciferol CRS,
colour and size to the spot in the chromatogram obtained
without heating, in 10.0 mL of taluene R and dilute to
with reference solution (a). The chromatogram obtained with 100.0 mL with the mobile phase.
reference solution (b) shows immediately at the same level an
orange principal spot, which gradually becomes reddish-
Reference solution (b) Dilute 1.0 mL of chole<alciferol for
brown and remains so for 10 min.
sysum suitability CRS to 5.0 mL with the mobile phase. Heat
in a water-bath at 90°C under a reflux condenser for 45 min
E. Ultraviolet and visible absorption spectrophotometry
and cool.
(2.2.25).
Reference solution (c) Dissolve 0.10 g of chole<alciferol CRS,
Testsdiuion Place 5.0 mL of me test solution prepared for without heating, in toluene R and dilute to 100.0 mL with the
the assay in a suitable flask and evaporate to dryness under same solvent.
reduced pressure by swirling in a water-bath at 40 °C. Cool
under running water and restore atmospheric pressure with
Reference solution (d) Dilute 5.0 mL of reference solution (c)
nitrogen R. Dissolve the residue immediately in 50.0 mL of to 50.0 mL with the mobile phase. Keep me soludon in iced
cydohexane R. water.
Spearal range 250-300 om. Reference solution (e) Place 5.0 mL of reference solution (c)
in a volumetric flask, add about 10 mg of
Absorption maximum At 265 om. butylhydroxytaluene R and displace the air from the flask with
C. Examine me chromatograms obtained in the assay. nitrogen R. Heat in a water-bath at 90°C under a reflux
Results The principal peak in me chromatogram obtained condenser, protected from light and under nitrogen R) for
with the test solution is similar in retention time to the 45 min. Cool and dilute to 50.0 mL with the mobile phase.
principal peak in the chromatogram obtained with reference Column:
solution (a). - size: I = 0.25 m, 0 = 4.6 mm;
TESTS - stationary phase: silica gelfor chromalOgraphy R (5 urn).
Related substances Mobile phase pentanol R, hexaneR (3:997 VIV).
The thresholds indicated under Related substances Flow rale 2 mUmin.
(Table 2034.-1) in the general monograph Subsranasfor Detection Spectrophotometer at 254 om.
phannaceutical use (2034) do not apply.
Injection The chosen volume of each solution (the same
ASSAY volume for reference solution (a) and for the test solution);
Cany out the assayas rapidly as possible, avoiding exposure to automatic injection device or sample loop recommended.
actinic light and air. Relative retention With reference to cholecalciferol: pre-
Liquid chromatography (2.2.29). cholecalciferol = about O.4j trans-cholecalciferol = about 0.5.
Testsolution Introduce into a saponification flask a quantity System suitability Reference solution (b):
of the preparation to be examined) weighed with an accuracy - resolution: minimum 1.0 between the peaks due to pre-
of 0.1 per cent, equivalent to about 100 000 IU. Add 5 mL cholecalciferol and lrans-cholecalciferol; if necessary,
of water R) 20 mL of anhydrous ethanol R) 1 mL of sodium adjust the proportions of the constituents and the flow
ascorbate solution Rand 3 mL of a freshly prepared rate of me mobile phase to obtain this resolution;
50 per cent mlm solution of potassium hydroxide R. Heat in a - repeatability: maximum relative standard deviation of
water-bath under a reflux condenser for 30 min. Cool rapidly 1.0 per cent for the peak due to cholecalciferol after
under running water. Transfer the liquid to a separating 6 injections.
funnel with the aid of 2 quantities, each of 15 ml; of Calculate the conversion factor (f) using the following
walei' R, 1 quantity of 10 mL of ethanol (96 per cen!! Rand expression:
2 quantities, each of 50 ml., of pentane R. Shake vigorously
for 30 s. Allow to stand until the 2 layers are clear. Transfer K-L
the lower aqueous-alcoholic layer to a 2 nd separating funnel M
and shake with a mixture of 10 mL of ethanol (96 percen!! R
and 50 mL of pentane R. After separation, transfer the K area (or height) of the peak due to cholecalciferol in the
chromatogram obtained with reference solution (d)j
aqueous-alcoholic layer to a 3 rd separating funnel and the
L area (or height) of the peak due to cholecalciferol in the
pentane layer to the 1st separating funnel, washing the chromatogram obtained with reference solution (e);
2 nd separating funnel with 2 quantities) each of 10 ml., of M area (or height) of the peak due to pre-cholecalciferol in the
pentane R and adding the washings to the 1st separating chromatogram obtained with reference solution (e).
funnel. Shake the aqueous-alcoholic layer with 50 mL of
pentane R and add the pentane layer to the I" funnel. Wash The value off determined in duplicate on different days may
the pentane layer with 2 quantities, each of 50 ml., of a be used during the entire procedure.
freshly prepared 30 giL solution of potassium hydroxide R in Calculate the content of cholecalciferol in International Units
ethanol (10 percent VII-? R, shaking vigorously, then wash per gram using the following expression:
with successive quantities, each of 50 rnl., of water R until
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1-678 Colestyramine 2022
m' V SD + if x S } inflated diameter of 3-6 ern (flat width of 5-9 em) in ssater R
V' x;; X S/ P x40000x 1000
D to hydrate until pliable, appropriately sealing one end.
Introduce 2.0 g of the substance to be examined into the
'n mass of me preparation [0 be examined in me test solution,
in milligrams; tube and add 10 mL of water R. Seal the tube and
m' mass of ,hoI=l.;iJero/ eM in reference solution (a), in completely immerse it in 100 mL of water R in a suitable
mmigr.uns; vessel and stir the liquid for 16 h to effect dialysis. Use the
V volume of the test solution (25 mL); dialysate as test solution.
V' volume of reference solution (a) (100 mL);
So area (or height) of the peak due to cholecalciferol in the Reference solution Prepare the reference solution in a similar
chromatogram obtained with me rest solution; manner but using 10 mL of a freshly prepared 0.1 gIL
S' o area (or height) of me peak due to cholecalciferol in the solution of benzyltrimethylammonium chloride R instead of the
chromatogram obtained wim reference solution (a);
Sp area (or height) of the peak due to pre-choleealejfeeol in the
chromatogram obtained with the tell solution; Transfer 5.0 mL of the test solution to a separating funnel
f conversion factor. and add 5 mL of a 3.8 gIL solution of disodiwn tetraborate R,
1 mL of a solution containing 1.5 gIL of bromo thymol blue R
STORAGE and 4.05 gIL of sodium carbonate Rand 10 mL of
In an airtight, well-filled container, protected from light. chloroform R. Shake the mixture vigourously for 1 min, allow
The contents of an opened container are to be used as soon the phases to separate and transfer the clear organic layer to
as possible; any unused part is to be protected by an a 25 mL volumetric flask. Repeat the extraction with a
atmosphere of nitrogen. further 10 mL of chl()l'()joTm R, combine the organic layers
LABELLING and dilute to 25 mL with chlorofann R. Measure the
absorbance (1.2.25) of the solution at the absorption
The label states the number of International Unitsper gram.
maximum at 420 nrn, using as compensation liquid a
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph Ell
solution prepared in the same manner but using 5.0 mL of
water R instead of the test solution.
Repeat the operation using 5.0 mL of the reference solution.
Colestyramine **** The absorbance obtained with the test solution is not greater
** * than that obtained with the reference solution.
(ph. Bur. monograph 1775) ***** Impurity A
11041-12-6
Testsolution Shake 5.0 g with 10 mL of aawne R for
Action and use 30 min. Centrifuge and use the supernatant.
Lipid-regulating drug. Reference sduuon (a) Dissolve 5 mg of styrene R in acetone R
Preparation and dilute to 100.0 mL with the same solvent. Dilute 1.0 mL
Colestyramine Oral Powder of the solution to 100.0 mL with acetone R.
PIlE" _
Re/erenu solution (b) Dissolve 0.35 mL of styrene R in
acetone R and dilute to 100.0 mL with the same solvent.
DEFINITION Dilute 1.0 mL of the solution to 100.0 mL with aatone R.
Strongly basic anion-exchange resin in chloride form, Reference solution (e) Dissolve 0.35 mL of toluene R in
consisting of styrene-divinylbenzene copolymer with aatQne R and dilute to 100.0 mL with the same solvent.
quaternary ammonium groups. Re/erena solution (d) Mix 1.0 mL of reference solution (b)
Nominalexchang« capacity 1.8 g to 2.2 g of sodium and 1.0 mL of reference solution (c) with acetone Rand
glycocholate per gram (dried substance). dilute to 100.0 mL with the same solvent.
CHARACTERS Column:
Appearance - size: I:::::: 0.30 m, 0 : : : 3.9 mrn,
White or almost white, fine powder, hygroscopic. - stationary phase: oetadety/silyl silica gelfor chromawgraphy R
(10 urn) with a specific surface area of 330 m 2/g and a
Solubility
pore size of 12.5 nm.
Insoluble in water, in methylene chloride and in ethanol
(96 per cent). lHobi/e phase auwnitrile R, waterR (50:50 VIV).
Flow rate 2.0 mIJmin.
IDENTIFICATION
Comparison colestyramine CRS. Inj«tiotl 20 ~L of test solution and reference solutions (a)
and (d).
B. Chloride (see Tests).
TESTS - resolution: minimum 1.5 between the peaks due to
pH (2.2.3) impurity A and toluene.
4.0 to 6.0. Limit:
Suspend 0.100 g in 10 mL of warer R and allow to stand for - impurityA: Dot more than the area of the principal peak
10 min. in the chromatogram obtained with reference solution (a)
DiaIysable quaternary amines (1 ppm).
Maximum 500 ppm, expressed as benzyltrimethylarnmonium Chloride
chloride. 13.0 per cent to 17.0 per cent (dried substance).
Test selution Place a 25 em piece of cellulose dialysis tubing
having a molecular weight cut-off of 12 000-14 000 and an
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2022 Colistirnethate Sodium 1-679
To 0.2 g add 100 mL of waterR and so mg of potassium

nitrate R. Add, with stirring, 2 mL of niuic acid R and titrate Colistimethate Sodium
with 0.1 M silver nitrate, determining the end-point (Ph. Bur. monograph 0319)
potentiorne trically (2.2.20).
I mL of 0.1 M silvernitrate is equivalent to 3.55 mg of Cl.
Maximum 12 pee cent, determined on 1.000 g by drying in
vacuo at 70 °C at a pressure not exceeding 7 kPa for 16 h.
Na03S 1 r: S03 Na
ASSAY • 1i'\l1i'
Exchange capacity L-OAB=L-OAB or ~-OAB
Liquid chromatography (2.2.29). DAB =2.4-dlamlnobulanoic acid
Solution A Dissolve 1.500 g of sodium glycocholate R in a Polymyxin E1 deriva&le: R = CH3
solution containing 4 gIL of potassium dihydrogen phosphate R Polymyxin E2 derivalive: R =H
and 12 gIL of dipotassium hydrogen phosphate R and dilute to
Between 2 and 5 of the L-OAa residues are disubs~luled al N'
CMS E1ASM8: principal polymyxin E1 wilh 4 disubslitoled residues
Test so/mum Add 20.0 mL of solution A to a quantity of the
substance to be examined equivalent to about 0.100 g of the CMS E1ASM6: princlpal poIymyxill El wilh 3 dlsubsliluled residues
dried substance. Shake mechanically for 2 h and centrifuge CMS E1ASM4: principal polymyxin E1 wiIh 2 disubsUluled residues
for 15 min. Dilute 5.0 mL of the supernatant to 50.0 mL CMS E2ASM8: priodpal polymyxin E2 wiIh 4 disubslilUled residues
with water R.
CMS E2ASM6: principal polymyxin E2 with 3 disubsliluted residues
Reference solution (a) Dilute 4.0 mL of solution A to
CMS E2ASM4: principal polymyxin E2 wilh 2 dlsubsliluled residues
Reference solution (b) Dissolve 60 rng of sodium g/yctxholate R
and 30 mg of sodium taurodeoxychokue R in water Rand 8068-28-8
dilu re to 100 mL with the same solvent. Dilute 1 mL of the
solution to 10 mL with water R. Action and use
Antibacterial.
Column: .
- size; I;;;;; 0.25 m, 0 ;;;;; 4.6 mm, Preparations
- stationary phase: octadecylsilyl silica gel for chromatography R Colistimethate Inhalation Powder, hard capsule
(5 urn). Colistimethate Sodium for Injection
Mobile phase MiK 35 volumes of acetonitrile Rand Colistimethate Sodium Powder for Nebuliser Solution
65 volumes of a 10.9 gIL solution of potassium dihydrogen PhEIK _
phosphate R adjusted to pH 3.0 with phosphoric acid R.
Flow rate 1.5 mUmin. DEFINITION
Deteaion Spectrophotometer at 214 om. Colistimethate sodium is prepared from colistin by the action
of formaldehyde and sodium hydrogen sulfite to form a
Injeaion 50 ~L. mixture of di to penta bis-sulfornethylated primary amine
Run time Twice the retention time of glycocholate. derivatives, mainly polymyxins EI and E2.
System suitability Reference solution (b): Semi-synthetic product derived from a fermentation product.
Content
glycocholate and taurodeoxycholate,
Minimum 11 500 IU/mg (dried substance).
Calculate the nominal exchange capacity using the following
expression: PRODUCTION
The composition and purity of the colistin starting material
(2.5 Al - A 2 ) x m. x 1.2 used in the synthesis of colistimethate sodium are equivalent
12.5xA. xm2 to that described in the monograph Colistin sulfate (0320).
Al area of the peak due 10 glycocholate in the chromatogram
In addition, the colistin starting material must have a
obtained with reference solution (a), polymyxin EI content between 50 per cent and 75 per cent
A. area of the peak due to glycocholate in the chromatogram and a polymyxin E2 content between 5 per cent and
obtained with the test solution, 20 per cent. in order to comply with the composition limits
m. mass, in miUigrams, of sodium g/yeoduNal4 R used in !he
for colistirnethate sodium.
preparation of solution A,
"'. mass, in milligrams, of the dried substance 10 be examined used CHARACTERS
in the prepara Lion of the test solution,
1.2 correction factor 10 convert the true exchange capacity to the
Appearance
conventionally used nominal exchange capacity. White or almost white, hygroscopic powder.
Solubility
STORAGE Very soluble in water. slightly soluble in ethanol
In an airtight container. (96 per cent), practically insoluble in acetone.
IMPURITIES IDENTIFICATION
Specified impuniies A. A. Examine the chromatograms obtained in the test for
A. styrene. composition.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PfrEur
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1-680 Colistimethate Sodium 2022
0.100-.---
0.090
0.080
0.070
0.060 ' 2 4
0.050 e
0.040
0.030
0.020, 6
3
0.010 :
0.000'
0.010
a 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 36 40 42 min
I. CMS E2t\SM8 3. CMS E2ASM6 5. CMS E2t\SM4

2. CMS Elt\SM8 4. CMS Elt\SM6 6. CMS EIASM4
Figure 0319.-1. - Chromatogram for the lestfor composuion of coIislimethau sodium: reference solution (a)
Results The peaks due to CMS EIASM8, CMS EIASM6, Reference solution (a) Dissolve 10 mg of colislimethate sodium
CMS EJASM4, CMS E2ASM8, CMS E2ASM6 and for PMk identification CRS in 0.25 mL of waterR and dilute
CMS E2ASM4 in the chromatogram obtained with the test to 5.0 mL with methanol R.
solution are similar in retention time to the corresponding Reference solution (b) Dissolve 2 mg of HI colistimethau
peaks in the chromatogram obtained with reference sodium for peak identification CRS in 0.25 mL of waterR and
solution (a). dilute (0 2.0 mL with methanol R.
B. It gives reaction (b) of sodium (2.3.1). Reference solution (c) Dissolve 1.5 mg of £2 coIistimethate
TESTS sodium for PMk identification CRS in 0.25 mL of water Rand
Appearance of solution dilute to 5.0 mL with methanol R.
The solution is clear (2.2.1). Reference solution (d) Dilute 1.5 mL of reference solution (a)
Dissolve 0.16 g in 10 mL of waterR. to 25.0 mL with methanolR.
pH (2.2.3) Precolumn:
6.5 to 8.5. - size: 1= 5 mm, 0 = 2.1 mm;
- stationary phase: end-capped oaadecy/silyl silica gelfor
chromatography R (1.7 11m).
10 mL with the same solvent. Measure after 30 min.
Column:
Free colIstln - size: 1= 0.15 m, 0 = 2.1 rom;
Dissolve 80 rng in 3 mL of waterR. Add 0.1 mL of a - stationary phase: end-eapped actadecy/si/yl silica gelfor
100 gfL solution of silicotrmgstic acidR; 10-20 s after addition chromatography R (1.7 urn),
of the reagent, the solution is not more opalescent than - temperature: 30 DC.
reference suspension Il (2.2.1).
Mobile phase:
Composition - mobile phase A: acetonitrile RI, buffer solution
Liquid chromatography (2.2.29): use the normalisation (25:475 V/JI);
procedure. - mobile phaseB: acetonitrile Rl, buffer solution
Buffersolution 7.8 gIL solution of sodium dihydrogen (250:250 VIJI);
pho$jJhate R adjusted to pH 6.5 with I M sodium hydroxide.
Test solution Dissolve 20 mg of the substance to be Time Mobile phase A Moblle phase B
(mln) (per cent VIP) (per cent VIP)
examined in 0.5 mL of waw Rand dilute to 10.0 mL with
methanol R. 0·10 80 .... 68 20 .... 32
10 - 35 68 .... 53 32 .... 47
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2022 Colistimethate Sodium 1-681
0.070;-
0.065'
0.060
0.055
0.050
0.045
0.040
0.035
0.030
0.025
0.020'
0.Q15
0.010'
:::!-1 U~~~_, "'- . . '- - ', ......,_~.-J

-o.OOS:
'-...-1
-0.010:
a 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 min
Figure 0319.-2. - Chromatogram for the tes: for composition of colisrimethate sodium: reference solution (b)
0.034'
0.032 1
0.030'
0.026
0.026:
0.024;
0.022:
0.020:
0.018
0.016'
0.014
0.0121
0.010;
0.006i
0.006,
0.004;
0.002
0.000
-0.002
-0.004
-0.006'
-0.008
------ ---:
-0.010' 42
o 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40' min
Figure 0319.-3. - Chromategram for the ten for composition of eolistimethaie sodium: reference solution (c)
Flow rate 0.30 mUmin. Jdentification of peaks Use the chromatogram supplied with
Detection Spectrophotometer at 210 run. colistimethate sodium for peak idemification CRS and the
Autosamplet Set at 5°C. identify the peaks due to CMS EIASM8, CMS EIASM6,
Injection 2.0 1-lL.
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1-682 Colistin Sulfate 2022
CMS EIASM4, CMS E2ASM8, CMS E2ASM6 and CMS Pyrogens (2.6.8)
E2ASM4 (see Figure 0319.-1). If intended for use in the manufacture of parenteral
The peak corresponding to the most abundant compound in preparations without a further appropriate procedure for
the range of 11.0-14.5 min (CMS EIASM6) in the removal of pyrogens, it complies with the test. Inject, per
chromatogram obtained with reference solution (a) is set as kilogram of the rabbit's mass, 1 mL of a solution in waterfor
the identification reference peak (relative retention 1.00). injections R containing 2.5 mg of the substance to be
examined per millilitre.
ldemijicatiml of peaks related eo CMS E1 and CMS E2 Use
the chromatogram supplied with E1 colisrimethate sodium for ASSAY
peak identificatWn CRS and E2 colistimethate sodium for peak Carry out the microbiological assay of antibiotics (2. 7.2).
identijication CRS, and the chromatograms obtained with Use colistimethate sodium CRS as the chemical reference
reference solutions (b) and (c) to identify all peaks related to standard.
CMS EI and CMS E2 in the chromatogram obtained with
STORAGE
the test solution (see Figures 0319.-2 and 0319.-3).
In an airtight container, protected from light. If the substance
Relative retention With reference to CMS BIASM6 is sterile the container is also sterile and tamper-evident
(retention time = about 13 min): CMS E2ASM8 = about PhEIl
=
0.22, CMS EIASM8 about 0.39; CMS E2ASM6 about = ~
=
0.71; CMS E2ASM4 about 1.77; eMS
=
EIASM4 about 2.35.
Integrate all peaks above 0.05 per cent to establish the total
area. Colistin Sulfate
Systemsuitability: Colistin Sulphate
the difference in the retention times of CMS E 1ASM6 in
-
2 consecutive injections of reference solution (a) is less
than 0.1 min; the drift in the retention time of CMS
EIASM6 from the beginning to the end of the sequence °
}-L.DAB- L-Thr- L-DAB- L·DAB- L-DAB- o-Leo- X J
is less than 0.5 min; R lL-Thr-L.DAB -L.DAB
- peak-eo-valley ratio: minimum 1.2, where H p = height
above the baseline of the peak with a relative retention of
=
about 2.37 and H" height above the baseline of the
/=~3 • R2 DAB =2,4-dlamlnobutanolc acid
lowest point of the curve separating this peak from the R4 '"R1
peak due to CMS BIASM4 in the chromatogram H
obtained with reference solution (a); polymyxin X Rl R2 R3 R4 Mol Formula M,
- number of theoretU:al plates: minimum 50 000, calculated
E1 l·Leu CH3 CH3 H H C53H,ooN1S013 1169
for the peak due to CMS EIASM6 in me chromatogram L-Leu H H
E2 CH3 H C52HgaNI60'3 1155
obtained with reference solution (a), E3 L-leu H CH3 H H C52HgeN16013 1155
- signal-eo-noise ratio: minimum 50 for the peak due to E4 t-Lsu H H H H Cs 1HgeN160'3 1141
CMS EIASM6 in the chromatogram obtained with E6 l-Leu CH3 C~ H OH C53HU:ON10014 1185
reference solution (d). E1·7MOA L-leu H CH3 CH3 H C53 HiooN,S013 1169
EH L·lle CH3 CH3 H H C53HIOON,s013 1169
Limits:
E1·Nva L-Nva CH3 C~ H H C 52HgeNl0013 1155
- CMS E1ASM8: 5.0 per cent to 9.5 per cent; E2·1 L-lie CH3 H H H C52~N,sOI3 1155
l
- CMS E1ASM6: 6.5 per cent to 9.5 per cent; E2-Val L-Val CH3 H H H Cs1HgeN'6013 1141
- CMS E1ASM4: 2.0 per cent to 5.0 per cent;
- CMS ElASM8: 05 per cent to 2.0 per cent,
_2'_3~_e_h_yd_ro_nE_l
X Mol. F()(mula M,
- CMS ElASM6: 0.5 per cent to 2.5 per cent,
- eMS E2ASM4: maximwn 1.5 per cent;
rI: 1167
_
CH3
- sum oj thepeaks related to CMS E1 and CMS E2:
minimwn 77.0 per cent; ····CH
H 3
- disregard limir. 0.50 per cent.
Related substances 1264-72-8
composition. Action and use
Limits: Antibacterial.
- any other impun'ty (any peak not related to CMS EI or Preparation
CMS E2): for each impurity, maximum 1.5 per cent; Colistin Tablets
- sum of impurities (sum of all peaks not related to CMS EI
PhEtI _
or CMS E2): maximum 5.5 per cent; _~_~ ~
- disregard limit: 0.50 per cent. DEFINITION

Loss on drying (2.2.32) A mixture of the sulfates of polypeptides produced by certain
Maximum 5.0 per cent, determined on 1.000 g by drying in strains of Bacillus polymyxa var. colistinus.
'Vacuo at 60°C at a pressure not exceeding 0.7 kPa for 3 h.
Content
Sulfated ash (2.4.14) Minimum 19 000 IU/mg (dried substance).
16 per cent to 21 per cent, determined on 0.50 g.
CHARACTERS
Appearance
While or almost white, hygroscopic powder.
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2022 Colistin Sulfate 1-683
Solubility Test solurion Dissolve 5.0 mg of the substance to be

Freely soluble in water, practically insoluble in acetone and examined in 8 mL of water R and dilute to 10.0 mL with
in ethanol (96 per cent). acetonitrile R.
IDENTIFICATION Reference solution (a) Dissolve 5.0 mg of colistin for system
First identification: B, E. suitability CRS in 8 mL of waterR and dilute to 10.0 mL
with acetonitrile R.
to 100.0 mL with a mixture of 20 volumes of acetonitrile R
Testsolution Dissolve 5 mg of the substance to be examined and 80 volumes of waterR.
in 1 mL of a mixture of equal volumes of hydroth/on"c acid R
Column:
and water R. Heat at 135°C in a sealed rube for 5 h.
- size: I ::::: 0.25 m, 0 = 4.6 nun;
Evaporate to dryness on a water-bath and continue the
- stationary phase: end-capped actadecy/silyl silica gelfor
heating until moistened blue litmus paperR does not tum red.
chromatography R (3.0 1"0);
Dissolve the residue in 0.5 mL of water R.
Reference soltuion (a) Dissolve 20 mg of leucine R in water R
l\tlobile phase Mix 22 volumes of acetonitrile Rl and
and dilute to 10 mL with the same solvent.
78 volumes of a solutionprepared as follows: dissolve 4.46 g
Reference solution (b) Dissolve 20 mg of threonine R in of anhydrous sodium su!fa,. R in 900 mL of waterfor
water R and dilute to 10 mL with the same solvent. chromatography R, adjust to pH 2.4 with dilu" phosphori<
Reference solution (c) Dissolve 20 mg of phel()'lalanine R in acid R and dilute to 1000 mL with waterfor
water R and dilute to 10 mL with the same solvent. chromatography R.
Reference sdution (d) Dissolve 20 mg of serine R in water R Flow rate 1.0 mUmin.
and dilute to 10 mL with the same solvent. Detection Spectrophotometer at 215 om.
PIa,. TLC silica gelplateR. Injection 20 IlL.
Cany out thefollowing procedures pro,.cled from light. Run time 1.5 times the retention time of polymyxin El.
MobIle phase waterR, phenol R (25:75 VIV). Identification of peaks Use the chromatogram supplied with
Application 5 ~L as bands of 10 mm, then place the plate in colistin for system suitability CRS to identify the peaks due to
the chromatographic tankso that it is not in contact with the polymyxins EI, E2, E3, E4, E6, EI-7MOA, EI-I, EI-Nva,
mobile phase, and allow it to become impregnated with the E2-I, E2-Val, 2,3-dehydro EI and due to impurities A
vapour of the mobile phase for at least 12 h. and B.
Development Over halfof the plate. Relative retention With reference to polymyxin HI (retention
Drying At 105 'C. time = about 21 min): polymyxin. E4 and
Detection Spray with ninhydrin solution Rl and heat at E2-Val = about 0.28; polymyxin E6 = about 0.39; polymyxin
110 -c for 5 min. E2-I = about 0.42; polymyxin E2 = about 0.50;
Results The chromatogram obtained with the test solution
= =
impurity A about 0.53; polymyxin E3 about 0.56;
polymyxin EI-Nva = about 0.59; polymyxin
shows zones corresponding to those in the chromatograms
obtained with reference solutions (a) and (b), but showsno
= =
EI-I about 0.82; polymyxin 2,3-<1ehydro EI about 0.90;
zones corresponding to those in the chromatograms obtained
=
polymyxin EI-7MOA about 1.1; impurity B about 1.3. =
with reference solutions (c) and (d); the chromatogram System suitability:
obtained with the test solution also shows a zone with a very - resolution: minimum 2.0 between me peaks due to
low R p value (2,4-diaminobutyric acid). polymyxin E6 and polymyxin E2-I, and minimum 3.0
between the peaks due to polymyxin 2,3-<1ehydro EI and
polymyxin El in the chromatogram obtained with
composition.
reference solution (a);
Results The peaks due to polymyxin El and polymyxin E2 - signal-to-noise ratio: minimum 10 for me peakdue to
in the chromatogram obtainedwith the test solution are polymyxin EI in the chromatogram obtained with
similar in retention time to the corresponding peaks in the reference solution (b);
chromatogram obtained with reference solution (a). - penk-UJ-vaJley ratio: minimum 1.1, where Hp = height
C. Dissolve about 5 mg in 3 mL of wa,.r R. Add 3 mL of above the baseline of the peak due to impurity A and
duu,. sodium hydroxide solution R. Shake and add 0.5 mL of a H; : : : height above the baseline of the lowestpoint of the
10 gIL solution of copper suffa,. pencahydra,. R. A violet curve separating this peak from the peak due to
colour is produced. polymyxin HZ in the chromatogram obtained with
D. Dissolve about 50 mg in I mL of 1 M hydrachlori< acid reference solution (a).
and add 05 mL of 0.01 M iodine. The solution remains Limits:
coloured. - correction factor: for the calculation of content, multiply the
E. It gives reaction (a) of sulfates (2.3.1). peak area of polymyxin 2,3-dehydro EI by 0.3;
. - polymyxin EI-I: maximum 8.5 per cent;
TESTS
- polymyxin E3: maximum 5.5 per cent;
pH (2.2.3) - polymyxin El-7MOA: maximum 5.0 per cent;
4.0 to 6.0. - polymyxin £6: maximum 4.5 per cent;
Dissolve 0.1 g in carbon dioxide-free waterR and dilute to - polymyxin El-Nua: maximum 4.5 per cent;
10 mL with the same solvent. - sum of poIymyxins E4 and E2-Val: maximum 3.0 per cent;
Composition - polymyxin £2-/: maximum 2.5 per cent;
Liquid chromatography (2.2.29): use the normalisation - polymyxin 2,3-dehydro EI: maximum 1.5 per cent;
procedure.
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1-684 Copovidone 2022
sum of polymyxim nt, E2, E3, E4, E6, Hl-7MOA, HI-I.

-
E1-Nva, E2-1. E2-Val and 2,3-Jehydro E1: minimum Copovidone1
86.0 per cent; (ph. Eur. monograph 0891)
- disregard limir: 0.35 per cent.
Related substances
composition.
Limits:
- any otherimpurily: for each impurity, maximum n= 102m
25 per cent, and not more than 4 such impurities exceed
1.0 per cent;
Mr (1l1.1)~ + (86.l)m 2508~-9
- ICtal: maximum 11.0 per cent;
- disregard limit: 0.35 per cent. Action and use
Sulfate Excipient in pharmaceutical products.
16.0 per cent to 18.0 per cent (dried substance). PhCII _
Dissolve 0.250 g in 100 mL of water R and adjust to pH 11
with concemrated ammonia R. Add 10.0 mL of 0.1 M barium DEFINITION
chloride and about 0.5 mg of phlhalein purple R. Titrate with Copolymer of 1-ethenylpyrrolidin-2-one and ethenyl acetate
0.1 M sodium edetate, adding 50 mL of ethanol (96 per cent) R in the mass proportion 3:2.
when the colour of the solution begins to change and Content
continuing me titration unti.l the violet-blue colour - nitrogen (N; A r 14.01): 7.0 per cent to 8.0 per cent (dried
disappears. substance);
1 mL of 0.1 M barium chloride is equivalent to 9.606 mg - elhenyl acetate (C 4H6 0 2; M r 86.10): 35.3 per cent to
of 804 . 42.0 per cent (dried substance).
Loss on drying (2.2.32) +CHARACTERS
Maximum 3.5 per cent, determined on 1.000 g by drying in Appearance
vacuo at 60 °C at a pressure not exceeding 0.1 kPa for 3 h. White or yellowish-white hygroscopic powder or flakes.
Sulfated ash (2.4.14) Solubility
Maximum 1.0 per cent, determined on l.0 g. Freely soluble in water, in ethanol (96 per cent) and in
ASSAY methylene chloride. +
Carry out the microbiological assay of antibiotics (2.7.2). IDENTIFICATION
Use cdistin sul/ate for microbic/ogical assay CRS as the First identification: A.
chemical reference substance. OSewndidentification: B, C.O
STORAGE A. Infrared absorption spectrophotometry (2.2.24).
In an airtight container, protected from light. Preparation Dry the substance to be examined and the
IMPURITIES reference substance at lOS °C for 3 h.
Specified impuniies B. Comparison copooidone CRS.
Otherdetectable impun'ties (the following substance« would, if {lB. To 1 mL of solution S (see Tests) add 5 mL of water R
present at a sufficient level, be detected by one or other0/ the leStf and 0.2 mL of 0.05 M iodine. A red colour appears.
in the monograph. They arelimited by thegeneral acceptance C. Dissolve 0.1 g of hydroxy/amine hydrochloride R in to mL
criterion for other/unspecified impurities. It is therefore not of methanol R, add 20 mL of a 40 gIL solution of sodium
necessary to identifythese impurities for demonstration of hydroxide R and filter if necessary. To 5 mL of the solution
compliance. See also 5./0. Control 0/ impuritief in subuances for add 0.1 g of the substance [0 be examined and boil for
phamulCeutical use) A. 2 min. Transfer 50 j.tL to a filter paper and add 0.1 mL of a
A. unknown structure, mixture of equal volumes of/emc chloride solution RI and
hydrochlorU:. acid R. A violet colour appears.e
O~_DAB_l_Thr_L_DAB_L_DABQAB_D-LeU-l_LeUl TESTS
ll-T/V ---t-DAB -L-DAB...J Solution S
CH3 ,. Dissolve 10.0 g in water R and dilute to 100.0 mL with the
"·'CH same solvent. Add the substance to be examined to the
H 3
water R in small portions with constant stirring.
Solution 8 is not more opalescent than reference
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIciI'
suspension ill (2.2.1) and not more intensely coloured than
reference solution B5 , R.; or BY 5 (2.2.2, Method11).
pH (2.2.3)
3.0 [0 7.0 for solution S.
I Thismonograph has undergone pharmacopoeia} harmoniuuion;
Seechapt8 5.8. Pharmawpoeial harmonisation:
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2022 Copovidone 1-685
ViscosIty) expressed as K..value trichloride-sulfuric acid reagent R J mix and allow to stand for
Dilute 5.0 mL of solution S to 50.0 mL with waterR. Allow 30 min. The absorbance (2.2.25) of the solution. measured
to stand for I hand determine the viscosity (2.2.9) of the at 405 nm using a mixture of 25.0 mL of the stock solution
solution at 25 ± 0.1 °C. Calculate the K-value using the and 2.0 mL of a J 3 per cent VIV solution of sulfuric acid R as
following expression: the compensation liquid, is not greater than 0.35.
Hydrazlne
1.510g'~1 - I V300clog.", + (c + 1.5c1og .",)' Thin-layer chromatography (2.2.27). UsefreshIY prepared
3 c'
0.15 + 0.003c + -'--------OO:-;.1c:5:-;c-:-+-::0'-;.0C:0:::-' -- - solutions.
c concentration of the substance to be examined (dried
Tesl solution Dissolve a quantity of the substance to be
substance). lIt gmms per 100 m£.; examined equivalent to 2.5 g of the dried substance in
lInf kinematic viscosity of the solunon relative [0 that of flHlter R. 25 mL of waterR and mix. Add 0.5 mL of a 50 gIL solution
of salicylaldehyde R in methanol R, mix and heat in a water-
The K-value is 90.0 per cent to 110.0 per cent of the bath at 60°C for 15 min. Allow to cool, add 2.0 mL of
nominal K-value. toluene R, stopper tightly, shake vigorously for 2 min and
Aldehydes centrifuge. Use the upper layer.
Maximum 500 ppm) expressed as acetaldehyde. Reference solution Dissolve 90 mg of salicylaldehyde azine R
Test solution Dissolve 1.0 g of the substance to be examined in toluene R and dilute to 100.0 mL with the same solvent.
in phosphate buffersolution pH 9.0 R and dilute to 100.0 mL Dilute 1.0 mL of the solution to 100.0 mL with toluene R.
with the same solvent. Stopper the flask tightly and heat at Plate TLC silanised SIlica gel F m plate R.
60°C for 1 h. AUow to cool to room temperature. Mobile phase waterR, methanol R (1:2 Vfl').
Reference solution Dissolve 0.140 g of acetaldehyde ammonia Application 10 ~L.
trimer trihydrate R in waterR and dilute to 200.0 mL with the Development Over 3/4 of the plate.
same solvent. Dilute 1.0 mL of the solution (0 100.0 mL
Drying In air.
with phosphate buffersolution pH 9. 0 R.
Into 3 identical spectrophotometric cells with a path length
Deteaion Examine in ultraviolet light at 365 om.
of 1 em, introduce separately 0.5 mL of the test solution, Retardation factor Salicylaldehyde azine = about 0.3.
0.5 mL of the reference solution and 0.5 mL of water R Limti:
(blank). To each cell add 2.5 mL of phosphate buffer solution - hydrazine: any spot due to salicylaldehyde azine is not
pH 9.0 Rand 0.2 mL of nicotinamide-adenine dinucleotide more intense than the spot in the chromatogram obtained
solution R. Mix and stopper tightly. Allow to stand at with the reference solution (l ppm).
22 ± 2°C for 2-3 min and measure the absorbance (2.2.25) ImpurltyA
of each solution at 340 nm, using water R as the Liquid chromatography (2.2.29).
compensation liquid. To each cell, add 0.05 mL of aldehyde
dehydrogenase solution R, mix and stopper tightly. Allow to
examined in 5 mL of methanol R. Sonicate until dissolution is
stand at 22 ± 2 °C for 5 min. Measure the absorbance of
complete and dilute to 100.0 mL with water R.
each solution at 340 om using water R as the compensation
liquid. Reference solntum Dissolve 0.150 g of 2-pyrrolidDne R
(impurity A) in the mobile phase and dilute to 100.0 mL
Calculate the content of aldehydes, expressed as
with the mobile phase. Dilute 3.0 mL of the solution to
acetaldehyde, in parts per million, using the following
expression:
Precolumn:
f(A:;-,,'------=::Ac:-,'~)_--7(Ac:;=-,,_-....:A~b"f') x 100 000 x C - size: 1:;;; 0.010 m, 0 :;;; 4 mm;
(A" - A,,) - (Ab> AbI) m - stationary phase: base-deactivated end-eapped octatlecylsilyl
silica gelfor ehromawgraphy R (5 urn).
All absorbance of the test solution before the addition of aldehyde
dehydrogenase;
Column:
Aa absorbance of the test solution after the addition of aldehyde =
- size: / 0.15 m, 0 :;;; 4.6 rnm;
dehydrogenase; - stationary phase: base-deactivated end-eapped octadecylsilyl
A'l absorbance of the reference solution before the addition of silica gelfor chromawgraphy R (5 urn),
aldehyde dehydrogenase;
A,z absorbance of the reference solution after the addition of
aldehyde dehydrogenase; Mobile phase methanol R2, waterfor chromawgraphy R
A bl absorbance of the blank before the addition of aldehyde (5:95 VIV).
dehydrogenase;
An absorbance of the blank after the addition of aldehyde FWto rate 0.8 mllmin.
dehydrogenasej Detection Spectrophotometer at 205 nm.
m mass of the substance to be examined (dried substance) in the
test solution, in gnunSj Injution 20 pL. After each injection of the test solution,
C concentration of acetaldehyde in the reference solution. elute and wash away the remaining sample by passing the
calculated from the mass of acetaldehyde ammonia trimer mobile phase through the column backwards for about
trihydrnte and by multiplying by a factor of 0.72. in milligrams
30 min at the same flow rate as applied in the test. This
per millilitte.
process may be replaced by washing the precolumn only.
Peroxides Run lime 4 times the retention time of impurity A.
Maximum 400 ppm, expressed as H,O,. ldendficosion of impurities Use the chromatogram obtained
Dissolve a quantity of the substance to be examined with the reference solution to identify the peak due to
equivalent to 4.0 g of me dried substance in water Rand impurity A.
dilute to 100.0 mL with the same solvent (stock solution). Retention lime Impurity A :;;; about 7 min.
To 25.0 mL of the stock solution add 2 mL of titanium
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1-686 Copovidone 2022
Systemsuitability Reference solution: - repeatability: maximum relative standard deviation of

- symmetry factor: maximwn 1.5 for the peak due to 2.0 per cent for the areas of the peaks due to impurities B
impurity A; and C determined on 6 injections. Use separate vials for
- repeatability: maximum relative standard deviation of each replicate injection.
2.0 per cent determined on 6 injections. Calculate the content of impurity B in parts per million using
Calculate me percentage content of impurity A using the the following expression:
Alxm2x50
A2 mj
Al area of the peak due to impurity B in the chromatogram

AI area of the peak due to impurity A in lite chromatogram
A2 area of the peak due to impurity B in rhe chromatogram
obtained with the reference solution;
A2 area of the peak due to impurity A in the chromatogram
>HI mass of the substance to be examined (dried substance) in the
obtained with the reference solution; test solution, in grams;
nIl mass of the substance to be examined (dried substance) in the
m2 mass of impurity B in the reference solution, in grams.
test solution, in grams;
nI2 mass of impurity A in the reference solution, in grams.
Calculate the content of impurity C in pans per million using
Limit: the following expression:
- impurity A: maximum 0.5 per cent
Impurities Band C
Liquid chromatography (2.2.29). Store the test solution and the
reference solution at a temperature not exceeding 10 °C and use A, area of the peak due to impuril}' C in the chromatogram
them within 8 h.
At area of the peak due to impurity C in the chromatogram
Test solution Dissolve 0.250 g of the substance to be obtained with the reference solution;
examined in the mobile phase and dilute to 10.0 mL with IN, mass of lhe substance to be examined (dried subs tance) in the
the mobile phase. test solution, in grams;
m2 mass ofimpurity C in the reference solution, in grams.
Reference solution Dissolve 50.0 mg of 1-'lJiny/pyrroridin-2-
one R (impurity B) and 50.0 mg of vinyl acetate R Limits:
(impurity C) in methanol R and dilute to 100.0 mL with the - impurities B, C: for each impurity, maximum 10 ppm.
with methanol R. Dilute 5.0 mL of this solution to 100.0 roL
Maximwn 5.0 per cent, determined on 0.500 g by drying in
Precolumn:
- size: / =0.033 m, 0 = 4.0 mrn;
- stationary phase: base-deactivated end-tapped Octadecy/silyl
silica gel for chromarography R (5 IJm). ASSAY
Column: Ethenyl acetate
- size: 1= 0.25 rn, 0 = 4.0 mm; Determine the saponification value (2.5.6) on 2.00 g of the
- stationary phase: base-deactivated end-eapped ocuulecylsilyl substance to be examined. Multiply the result obtained by
silica gel for chromatography R (5 IJm); 0.1534 to obtain the percentage content of the ethenyl
- temperature: 40 °C. acetate component.
Mobile phase acetonilrile R1, waterfor chromatography R Nitrogen
(8:92 VIV). Place 0.100 g of the substance to be examined (m mg) in a
Flow rate 1.0 mUmin. combustion flask and add 5 g of a mixture of 1 g of copper
sulfate pentahydmte R, 1 g of tiumium dioxide R and 33 g of
Detection Spectrophotometer at 205 om for impurity C and
dipotassium sulfau R. Wash any adhering particles from the
at 235 nm for impurity B.
neck into the flask with a small quantity of water R.
Injection 20 IJL. Mer each injection of the test solution, Add 7 mL of sulfuric acidR, allowing it to run down the
elute and wash away the remaining sample by passing the insides of the flask. Heat the flask gradually until the solution
mobile phase through the column backwards for about has a clear, yellowish-green colour, and me inside wall of the
30 min at the same flow rate as applied in the test. This flask is free from any carbonised material, and then heat for a
process may be replaced by washing the precolumn only. further 45 lnin. After cooling, add cautiously 20 mL of
Store and inject the solutions at a temperature not exceeding water R, and connect the flask to the distillation apparatus,
5 °C and use them within 8 h. Use a cooled autosarnpler, which has been previously washed by passing steam through
Run time Twice the retention time of impurity C. it. To the absorption flask add 30 mL of a 40 gIL solution of
Idendfication oj impurities Use the chromatogram obtained boric acid RJ 3 drops of bromocresol green-merhyl redsdution R
with the reference solution to identify the peaks due to and sufficient water R to immerse the lower end of the
impurities B and C. condenser tube. Add 30 mL of strong sodium hydroxide .
Retention time Impurity B = about 17 min; soluuon R through the funnel, rinse the funnel cautiously with
impurity C = about 22 min. 10 mL of water R, immediately close the clamp on the
rubber tube, then stan distillation with steam to obtain
80-100 mL of distillate. Remove the absorption flask from
- rew/ution: minimum 2.0 between the peaks due to
the lower end of the condenser tube, rinsing the end part
impurities Band C when the chromatogram is recorded
with a small quantity of water RJ and titrate the distillate with
at 205 nm;
0.025 M sulfuric acid until the colour of the solution changes
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2022 Copper Sulfate 1-687
from green through pale greyish-blue to pale greyish reddish-

purple. Carry out a blank determination,
Copper Sulfate
1 mL of 0.025 M sulfuric acid is equivalent to 0.700 mg ofN. Anhydrous Copper Sulfate
+STORAGE Anhydrous Copper Sulphate
In an airtight container.e (Ph. Eur. monograph 0893)
LABELLING CUS04 159.6 7758-98-7
The label states the nominal K-value.
Action and use
IMPURITIES Used in treatment of copper deficiency.
Specified impun'lies A, B, C.
PhErr _
DEFINITION
Content
A. pyrrolidin-2-one (2-pyrrolidone),
CHARACTERS
(CH2 Appearance
Greenish-grey powder, very hygroscopic.
(yo Solubility
Freely soluble in water, slightly soluble in methanol,
B. l-ethenylpyrrolidin-z-one (l-vinylpyrrolidin-2-one),
IDENfIFICATION
A. Add several drops of d.1ure ammonia R2 to I mL of
solution S (see Tests). A blue precipitate is formed.
On further addition of dilute ammonia R2 the precipitate
C. ethenyl acetate (vinyl acetate). dissolves and a dark blue colour is produced.
OFUNCTIONALITY-RELATED CHARACTERISTICS B. Loss on drying (see Tests).
This section provides infonnatUJn on characteristia that aw C. Dilute I mL of solution S to 5 mL with waterR.
recognised as being relevant control parameters for one or mow The solution gives reaction (a) of sulfates (2.3.1).
functions of the substance'when used as an excipient (see chapler TESTS
5.15). Some of the charactetistics descn'bed in the Functionality- Soludon S
maud characteristics section me{}' alsobe presen: in the mandatory Dissolve 1.6 g in water R and dilute to 50 mL with the same
part of the monograph since they also represen: mandawry quality solvent.
criteria; In such cases, a cross-reference to the tests descnoed in the
mandawrypart is included in the Funeticnality-relaud Appearance of solution
charaaeristics section. Controlof the characteristics can contribute Solution S is clear (2.2.1).
to the qualityof a medicinal product by improving the ccnsistency Chlorides (2.4.4)
of the manufactun'ng process and the perfonnana ofthe medicinal Maximum 150 ppm.
product during use. Whew control methods aw cited, they aw Dilute 10 mL of solution S to 15 mL with water R.
wcognised as being suiwblefor the purpose, but othermethods can Iron
alsobe used. Wherever results for a particular characteristic aw Maximum 150 ppm.
reported, the control methodmust be indicated.
The folktwing characieristics may be relevant for copovidone used
as binderin tablets and granules. Test solution Dissolve 0.32 g in 10 mL of waterR, add
2.5 mL of lead-free nitric acid R and dilute to 25.0 mL with
Viscosity (2.2.9) water R.
Determine the dynamic viscosity using a capillary viscometer
Referenu solutions Prepare the reference solutions using iron
on a 10 per cent solution (dried substance) or on a
standard solution (20 ppm Fe) R, adding 2.5 mL of lead-free
20 per cent solution (dried substance) at 25°C. It is typically
nitric acid R and diluting to 25.0 mL with water R.
about 8 ml'a-s or about 23 rnl'a-s, respectively.
Particle-size distribution (2.9.31 or 2.9.38)
Bulk and tapped density (2.9.14)
The following characteristic may be relevant for eopooidone used
as film former in coaleddosage forms and in aerosols. C!WJer m~ form explosive ace{Yfides with acetylene, Therifore,
clean the burnerthoroughly before any residues become dry.
Viscosity (2.2.9)
See above.O Lead
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph€Uf Maximum 5 ppm.
Atomic absorption spectrometry (2.2.23, Method 1)..
Test solution Dissolve 1.6 gin 10 mL of waterR, add
2.5 mL of lead-free nitric acidR and dilute to 25.0 mL with
warer R.
Reference solutions Prepare me reference solutions using lead
standard solution (100 ppm Ph) R, adding 2.5 mL of lead-free
nitric acid R and diluting to 25.0 mL with warer R.
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1-688 Copper Sulfate 2022
Source Lead hollow-cathode lamp. Testsolution Dissolve 0.5 gin 10 mL of waterR, add
Wavelengrh 217.0 nm. 2.5 mL of lead-fre« nitric acidR and dilute to 25.0 mL with
Atomisotion device Air-acetylene flame. waterR.
Copper me{}' form explosive aatylides with acetylene. Therifore, Referenu solutions Prepare the reference solutions using iron
clean the burner thoroughly before att}' residues become dry. standard solution (20 ppm Fe) R, adding 2.5 mL of lead-free
nitric acidR and diluting to 25.0 mL with waterR.
an oven at 250 ± 10°C. Wavelength 248.3 nm.
Awmuation device Air-acetylene flame.
ASSAY
Dissolve 0.125 g in 50 mL of water R. Add 2 mL of sulfurU; Copper mqy form explosive acetylides with acetylene. Therefore,
acid Rand 3 g of potassium iodide R. Titrate with 0.1 M clean the burner thoroughly before any residues become dry.
sodium thiosulfate, using 1 mL of starch solution R, added Lead
towards the end of the titration. Maximum 50 ppm.
1 mL of 0.1 M sodium thiosulfate is equivalent to 15.96 mg of Atomic absorption spectrometry (2.2.23, Method 1).
CuSO". Testsolution Dissolve 2.5 gin 10 mL of waterR, add
STORAGE 2.5 mL of lead-free nitric acidR and dilute to 25.0 mL with
In an airtight container. waterR.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _-PflEIl Referetlusolutions Prepare the reference solutions using lead
standard solution (100 ppm Ph) R, adding 2.5 mL of lead-free
nioic acidR and diluting to 25.0 mL with waterR.
Source Lead hoUow-cathode lamp.
Copper Sulfate Pentahydrate Wavelength 217.0 nm.
Copper Sulphate Pentahydrate Copper may form explosive autylideswith acetylene. Therefore,
(Ph. Bur. monograph 0894) clean the burner thoroughly before any residues become dry.
CuS04)5H20 249.7 7758-99-8 Loss on drying (2.2.32)
Action and use drying in an oven at 250 ± 10 °C.
Used in treatment of copper 'deficiency.
ASSAY
PhEtr _
Dissolve 0.200 g in 50 rnL of waterR. Add 2 mL of sulfuric
DEFINTIlON acid Rand 3 g of potassium iodide R. Titrate with 0.1 M
Content sodium thiosulfate, adding 1 mL of starch solution R towards
99.0 per cent to 101.0 per cent. the end of the titration.
I rnL 0.1 M sodium thiosulfate is equivalent to 24.97 mg of
CHARACTERS CuS04,5H20.
Appearance _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEm
Blue, crystalline powder or transparent, blue crystals.
Solubility
Freely soluble in water, soluble in methanol, practically
insoluble in ethanol (96 per cent).
Cortisone Acetate •••
* **
IDENTIFICATION ***••11
A. Add several drops of dilute ammonia R2 to 1 mL of (ph. Bur. monograph 0321)
solution S (see Tests). A blue precipitate is formed.
On further addition of dilute ammonia R2 the precipitate
dissolves and a dark blue colour is produced.
B. Loss on drying (see Tests).
C. Dilute 1 mL of solution S [Q 5 mL with water R.
The solution gives reaction (a) of sulfates (2.3.1).
TESTS
Solution S 402,5 50-04-4
Dissolve 5 g in waterR and dilute to 100 mL with the same
solvent. Action and use
Appearance of solution Corticosteroid.
Solution S is clear (2.2.1). Preparation
Chlorides (2.4.4) Cortisone Tablets
Maximum 100 ppm. PflEtr _
Dilute 10 mL of solution S to 15 mL with waw R.
DEFINITION
Iron
17-Hydroxy-3, 11,20-trioxopregn-4-en-21-yl acetate.
Maximum 100 ppm.
Atomic absorption spectrometry (2.2.23, Me/hod 1). Content
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2022 Cortisone Acetate 1-689
CHARACTERS Reference solution (a) Dissolve 25 mg of cortisone acetate CRS

Appearance in methanol R with gentle heating and dilute [0 5 mL with
White or almost white, crystalline powder. the same solvent (solution B). Dilute 2 mL of this solutionto
Solubillty 10 mL with methylene chloride R.
Practically insoluble in water, freely soluble in methylene Reference solution (b) Transfer 2 mL of solution B to a
chloride, soluble in dioxan, sparingly soluble in acetone, 15 mL glass tube with a ground-glass stopper or
slightly soluble in ethanol (96 per cent) and in methanol. a poljtetrafluoroethylene cap. Add 10 mL of saturated
It shows polymorphism (5.9). methanolic potassium hydrogen carbonate solution Rand
immediately pass a stream of nitrogen R briskly through the
IDENTIFICATION solutionfor 5 min. Stopper the tube. Heat in a water-bath at
First identification: A, B. 45 °C protected from lighr for 2.5 h. Allow to cool.
Secondidentification: CJ D, E. Plate TLC silica gel F254 plate R.
A. Infrared absorption spectrophotometry (2.2.24). Mobile phase Add a mixture of 1.2 volumes of water Rand
Comparison cortisone acetate CRS. 8 volumes of methanol R to a mixture of 15 volumes of
If the spectra obtained in the solid stateshow differences, erher Rand 77 volumes of methylene chloride R.
record new spectra using 50 gIL solutions in methylene Application 5 tJL.
chloride R in a 0.2 mm cell. Development Over a path of J5 em.
B. Thin-layer chromatography (2.2.27). Drying In air.
Solvem mixture methanol R, melhylene chlon'de R (1:9 V/II). Detection A Examine in ultraviolet light at 254 nm.
Test solution Dissolve 10 mg of the substance to be Results A The principal spot in each of the chromatograms
examined in the solventmixture and dilute £0 10 mL with obtainedwith the test solutions is similar in position and size
the solvent mixture. to the principal spot in the chromatogram obtainedwith the
corresponding reference solution.
Reference solution (a) Dissolve 20 mg of cortisone acetate CRS
in the solvent mixture and dilute to 20 mL with the solvent Detection B Spray with akoholi< solution of sulfurU; acid R and
mixture. heat at 120°C for 10 min or until the spots appear. AlJow to
Reference solution (b) Dissolve 10 mg of hydrocortisone cool. Examine in daylight and in ultraviolet light at 365 run.
acetate R in reference solution (a) and dilute to 10 mL with Results B The principal spot in each of the chromatograms
reference solution (a). obtainedwith the test solutions is similar in position, colour
Plare ttc silica gel F m plate R. in daylight, fluorescence in ultraviolet light at 365 nm and
size to the principal spot in the chromatogram obtained with
Mobilephase Add a mixture of 1.2 volumesof water Rand the corresponding reference solution. The principal spots in
8 volumes of methanol R to a mixture of IS volumes of the chromatograms obtainedwith test solution (b) and
etherRand 77 volumes of methylene chloride R. reference solution (b) have an Rp valuedistinctly lower than
Appli<ation 5 ~L. that of the principal spots in the chromatograms obtained
Development Overa path of 15 em. with test solution (a) and reference solution (a).
Drying In air. D. Add about 2 mg to 2 mL of sulfuric acid R and shake to
Detecdon A Examine in ultraviolet light at 254 nm. dissolve. Within 5 min, a faint yellow colour develops.
Results A The principal spot in the chromatogram obtained Add this solution to 10 mL of water R and mix. The colour
with the test solution is similar in positionand size to the is discharged and a clear solution remains.
principal spot in the chromatogram obtained with reference E. About 10 mg gives the reaction of acetyl (2.3.1).
solution (a).
TESTS
Detection B Spray with akoholi< sDiU/ion of sulfurU; acidR. Specific optical rotadon (2.2.7)
Heat at 120 °C for 10 min or until the spots appear. Allow + 211 10 + 220 (dried substance).
to cool. Examine in daylight and in ultraviolet light at
Dissolve 0.250 gin diaxan R and dilute to 25.0 mL with the
365 om.
same solvent.
with the test solution is similar in position, colour in daylight, Related substances
fluorescence in ultraviolet light at 365 nm and size to the Liquid chromatography (2.2.29). Prepare rhe solutions
principal spot in the chromatogram obtained with reference immediauly before use.
solution (a). Test solution Dissolve 25.0 mg of the substance to be
System suitability Reference solution (b): examined in acetonitrile R and dilute to 10.0 mL with the
- the chromatogram shows 2 clearly separated spots. same solvent.
C. Thin-layer chromatography (2.2.27). Reference solution (a) Dissolve 2 mg of cortisone acetate CRS
and 2 mg of hydrocortisone acetate CRS (impurity A) in
Test solution (a) Dissolve 25 mg of the substance to be
aawnitrile R and dilute to 100.0 mL with the same solvent.
examined in methanol R with gentleheating and dilute to
5 mL with the same solvent (solution A). Dilute 2 mL of this Reference solution (b) Dilute 1.0 mL of the test solution to
solution to 10 mL with methylene chlon·de R. 100.0 mL with acetonitrile R.
Test solution (b) Transfer 2 mL of solution A to a 15 mL Column:
glass tube with a ground-glass stopper or - size: I;;;; 0.25 m, 0 = 4.6 mm;
a polytetrafluoroethylene cap. Add 10 mL of saturated - stationary phase: octade<y/silyl silica gelfor chromatagraphy R
methanolic potassium hydrogen carbonate solution Rand (5 urn).
immediately pass a stream of mirogen R briskly through the Moinle phase In a 1000 mL volumetric flask mix 400 mL of
solution for 5 min. Stopper the tube. Heat in a water-bath at acetonitrile R with 550 mL of water R and allow to
45 °C protected from light for 2.5 h. Allow to cool. equilibrate; dilute to 1000 mL with warer R and mix again.
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1-690 Cottonseed Oil 2022
Flow rate 1 mUmin. Solubility

Detection Spectrophotometer at 254 nm. Practically insoluble in water, freely soluble in methylene
Equilibratwn With the mobile phase for about 30 min. chloride and in toluene, very slightly soluble in ethanol
Injection 20 ~L; inject acetonitrile R as a blank. (96 per cent).
Run time Twice the retention time of cortisone acetate. IDENTIFICATION
Retention lime Impurity A = about 10 min; cortisone A. Melting point (see Tests).
=
acetate about 12 min. B. Composition of fatty acids (see Tests).
System suitability Reference solution (a): TESTS
- resolution: minimum 4.2 between the peaks due to Melting point (2.2.14)
impurity A and cortisone acetate; if necessary, adjust the 57 QC to 70 °C.
concentration of acetonitrile in the mobile phase.
Acid value (2.5.1)
Limits:
Maximum 0.5.
- impUlity A: not more than 0.5 times the area of the
principal peak in the chromatogram obtained with Dissolve 10.0 g in 50 mL of a hot mixture of equal volumes
reference solution (b) (0.5 per cent); of ethanol (96 per cent) R and toluene R, previously neutralised
- total: not more than 1.5 times the area of the principal with 0.1 M potassium hydroxide using 0.5 mL of
peak in the chromatogram obtained with reference phenolphthalein solution R1 as indicator. Titrate the solution
solution (b) (1.5 per cent); immediately while still hot.
- disregard limit: 0.05 times the area of the principal peak in Peroxide value (2.5.5, M~thod A)
the chromatogram obtained with reference solution (b) Maximum 5.0.
(0.05 per cent). Unsaponlfiable matter (2.5.7)
Loss on drying (2.2.32) Maximum 1.0 per cent, determined on 5.0 g.
Maximum 05 per cent, determined on 0500 g by drying in Alkallne impurities
an oven at 105 °C. Dissolve with gentle heating 2.0 g in a mixture of 1.5 mL of
ASSAY ethanol (96 per cent) R and 3 mL of toluene R. Add 0.05 mL
Dissolve 0.100 g in ethanol(96 per cent) R and dilute to of a 0.4 gIL solution of bromophenol blue R in ethanol
100.0 rnL with the same solvent. Dilute 2.0 mL of this (96 per cent) R. Not more than 0.4 mL of 0.01 M hydrochloric
solution to 100.0 mL with ethanol (96 per cent) R. Measure arid is required to change the colour to yellow.
the absorbance (2.2.25) at the absorption maximum at Composition of fatty acids
237 nm. Gas chromatography (2.4.22, Method A) with the following
Calculate the content of C:BH30 0 6 taking the specific modifications. Use the mixture of calibrating substances in
absorbance to be 395. Table 2.4.22.-3.
STORAGE Column:
Protected from light. - material: fused silica;
IMPURITIES =
- size: 1= 25 m, 0 0.25 mm;
- stationaryphase: cyanopropylpofysiwxane R (film thickness
0.2 lim).
Carner gas helium for chromatography R.
Split ratio 1:100.
Temperature:
- column: 180 QC for 35 min;
o - injection pon and detector. 250 °C.
A. 1113,17-dihydroxy-3,20-dioxopregn-4-en-21-yl acetate Detection Flame ionisation.
(hydrocortisone acetate). Composition of thefatty-acid fraction of the substance:
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PllEl.r
- saturatedfauy acidsof chain length less than C 14: maximum
0.2 per cent;
- myristic acid: maximum 1.0 per cent;
Hydrogenated Cottonseed Oil *** - palmitic acid: 19.0 per cent to 26.0 per cent;
*** *** - stearic acid: 68.0 per cent to 80.0 per cent;
(ph. Eur. monograph 1305) *** - oleic acid and isomers: maximum 4.0 per cent;
- linoleic acidand isomers: maximum 1.0 per cent;
Ph€1I' _
- arachidic acid: maximum 1.0 per cent;
DEFINITION - behenic acid: maximum 1.0 per cent;
Product obtained by refining and hydrogenation of oil -lignocenc acid: maximum 0.5 per cent.
obtained from seeds of cultivated plants of various varieties of STORAGE
Gossypium hirsutum L. or of other species of Gossypium. Protected from light.
The product consists mainly of triglyceridesof palmitic and __________ ~ PIIEtT
stearic acids.
CHARACTERS
Appearance
White or almost white mass or powder which melts to a
clear, pale yellow liquid when heated.
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2022 Crude Cresol 1-691
VS is equivalent to 0.080 mL of volatile bases. Not more

Cresol than 0.15% v/v of volatile bases is present.
Action and use Hydrocarbons and volatile bases
Antiseptic; antimicrobial preservative. The sum of the contents of hydrocarbon oil and volatile
bases, as determined in the tests for Hydrocarbons and for
DEFlNITION
Volatile bases, does not exceed 0.25% vlv.
Cresol is a mixture of cresols and other phenols obtained
from coal tar. Sulfur compounds
Place 20 mL in a small conical flask and over the mouth of
CHARACTERISTICS the flask fix a piece of filter paper moistened with a 10% wlv
An almost colourless to pale brownish yellow liquid. solution of leadttt) acetate. Heat the flask on a water bath for
Almost completely soluble in 50 volumes of water; freely 5 minutes. Not more than a light yellow colour is produced
soluble in ethanol (96%), in ether and in fixed and volatile on the filter paper.
oils. Non-volatile matter
IDENTIFICATION When evaporated on a water bath and dried at 105 0 , leaves
Shake 0.5 mL with 300 mL of water and filter. The filtrate not more than 0.1 % wtv of residue.
complies with the following tests. STORAGE
A. Add irrm(lll) chloride solution R1. A transient blue colour is Cresol should be protected from light. It darkens with age or
produced. on exposure to light.
B. Add bromine water. A pale yellow flocculent precipitate is
produced.
TESTS
Acidity Crude Cresol
A 2.0% wlv solution is neutral to bromocresol purple rolution.
Distillation range
Not more than 2% v/v distils below 1880 and not less than
80% vlv distils between 195 0 and 205 0 , Appendix V C.
Weight per mL
1.029 to 1.044 g, Appendix V G.
Hydrocarbons 108.1
Place 50 mL in a 500 mL round-bottomed flask, add
150 mL of 5M sodium hydroxide and 30 mL of waterand mix Action and use
thoroughly. Connect the flask to a splash-bulb and air- Antiseptic.
condenser about 60 ern long, with the end of the air- PhEII _
condenser fitting closely into the neck of a 250 ml, pear-
shaped separating funnel and passing well into the separating DEFINITION
funnel, which has a cylindrical graduated portion above the Mixture of 2-, 3- and 4-methylphenol.
stopcock. Fill the graduated portion of the separating funnel CHARACTERS
with water. Distil rapidly until 75 ml, of distillate has been Appearance
collected, cooling the separating funnel in running water if Colourless or pale brown liquid.
necessary. Allow the separating funnel to stand in a vertical
Solubility
position until separation is complete and draw off the
Sparingly soluble in water, miscible with alcohol arid with
aqueous liquid into a titration flask for use in the test for
methylene chloride,
Volatile bases.
AUow the separating funnel to stand for a few minutes, IDENTIFICATION
measure the volume of hydrocarbon oil in the graduated A. To 0.5 mL add 300 ml, of water R, mix and filter.
portion and wann, if necessary, to keep the oil in the liquid To 10 mL of the filtrate add I mL ofleme chloride
state. Subtract the volume of volatile bases in the solution R 1. A blue colour is produced.
hydrocarbon oil, as determined in the following test. B. To 10 mL of the filtrate obtained in identification test A,
Not more than 0.15% vlv of hydrocarbon oil is present. add I mL of bromine waterR. A pale yellow flocculent
Volatile bases precipitate is produced.
To the aqueous liquid reserved in the test for Hydrocarbons C. Relative density (see Tests).
add any aqueous liquid still remaining in the separating TESTS
funnel and neutralise, if necessary, with a.1M hydrochlaric add
Solution S
using phenolphthalein solution R1 as indicator. Titrate with 1M
To 2.5 g of the substance to be examined add 50 mL of
hydrochloric acid VS using methylorange solution as indicator. water R, shake for 1 min and filter through a moistened filter.
Wash the oil from the separating funnel into the titration
flask with water and again titrate with 1M hydrochloric add VS. Acidity or alkalinity
From the volume of additional 1M hydrochloric acid VS, To 10 mL of solution S add 0.1 mL of methylredsOlUtion R
calculate the proportion of volatile bases in the hydrocarbon and 0.2 rnl, of 0.01 M sodium hydroxide. The solution is
oil. From the total volume of 1M hydrochloric acid VS used in yellow. Add 0.3 mL of 0.01 M hydrtXhforU: acid. The solution
both titrations calculate the volume of volatile bases in the is red.
substance being examined. Each mL of 1M hydrtXhloric acid Relative density (2.2.5)
1.029 to 1.044.
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1-692 Croscarmellose Sodium 2022
Distillation range (2.2.11) Sodium chloride Place 5.00 g in a 250 mL conical flask, add
A maximum of 2.0 per cent VIV distils below 188°C and a 50 mL of waterRand 5 mL of strong hydrogen peroxide
minimum of 80 per cent VIV distils between 195°C and solution R and heat on a water-bath for 20 min, stirring
205 °C. occasionally to ensure total hydration. Cool and add 100 mL
Sulfur compounds of water Rand 10 mL of nitricacid R. Titrate with 0.05 M
Place 20 mL in a small conical flask. Over the mouth of the silver nitrate, determining the end-point potentiometrically
flask fix a piece of filter paper moistened with leadacetate (2.2.20) using a silver indicator electrode and a double-
solution R. Heat on a water-bath for 5 min. Not more than a junction reference electrode containing a 100 gIL solution of
light yellow colour is produced on the filler paper. pctassium nitrate R in the outer jacket and a standard filling
solution in the inner jacket, and stirring constantly,
Maximwn 0.1 per cent, I mL of 0.05 M silvernarate is equivalent to 2.922 mg of
NaCI.
Evaporate 2.0 g to dryness on a water-bath and dry at
100-105 °C for 1 h. The residue weighs not more than 2 mg. Sodiumglycclate Place a quantity of the substance to be
examined equivalent to 0.500 g of the dried substance in a
STORAGE 100 mL beaker. Add 5 mL of glacial acetic acid Rand 5 mL
Protected from light. of water R and stir to ensure total hydration (about 15 min).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEu- Add 50 mL of acetone R and I g of sodium chloride R. Stir for
several minutes to ensure complete precipitation of the
carboxymethylcellulose. Filter through a fast filter paper
impregnated with acetone R into a volumetric flask, rinse the
Croscarmellose Sodium 1 beaker and the filter with 30 mL of acewne R and dilute the
filtrate to 100.0 mL with the same solvent. Allow to stand for
(ph. Eur. monograph 0985) 24 h without shaking. Use the clear supernatant to prepare
the test solution.
Action and use
Prepare the reference solutions as follows: in a 100 mL
Excipient.
volumetric flask, dissolve 0.100 g of glycolic acid R, previously
PhEu- _ dried in a desiccator (2.2.32) at room temperature overnight,
in water R and dilute to 100.0 mL with the same solvent; use
DEFINITION
the solution within 30 days; transfer 1.0 mL, 2.0 mL,
Cross-linked sodium carboxyrnethylcellulose.
3.0 mL and 4.0 mL of the solution to separate volumetric
Sodium salt of a cross-linked, partly O-carboxymethylated flasks, dilute the contents of each flask 10 5.0 mL with
cellulose. waterR, add 5 mL of g/ado,l acetic add R, dilute to 100.0 mL
tCHARACTERS with acetone R and mix.
Appearance Transfer 2.0 mL of the test solution and 2.0 mL of each of
White or greyish-white, hygroscopic powder. the reference solutions to separate 25 mL volumetric flasks.
Solubility Heat the uncovered flasks for 20 min on a water-bath to
Practically insoluble in acetone, in anhydrous ethanol and in e1iminate acetone. Allow to cool and add 5.0 mL of
toluene.• 2,7-dihydrqxynaphthalene solution R to each "ask. Mix, add a
further 15.0 mL of 2,7-dihydroxynaphtha1ene solution Rand
IDENTIFICATION mix again. Close the flasks with aluminium foil and heat on a
A. Mix I g with 100 mL of a solution containing 4 ppm of water-bam for 20 min. Cool and dilute to 25.0 mL with
methylene blue R, stir the mixture and allow it to settle. ruifunc acid R.
The substance to be examined absorbs the methylene blue Measure the absorbance (2.2.25) of each solution at 540 om.
and settles as a blue, fibrous mass. Prepare a blank using 2.0 mL of a solution containing
B. Mix I g with 50 mL of water R. Transfer 1 mL of the 5 per cent VIV each of glacial acetic acid R and water R in
mixture to a small test-tube and add 1 mL of waterR and acewne R. Prepare a standard curve using the absorbances
0.05 mL of a freshly prepared 40 gIL solution of obtained with the reference solutions. From the standard
a-naphthol R in methanol R. Incline the test-tube and carefully curve and the absorbance of the test solution, determine the
add 2 mL of sulfuric acidR down the side so that it forms a mass (a) of glycolic add in the substance to be examined, in
lower layer. A reddish-violet colour develops at the interface. milligrams, and calculate the content of sodium glycolate
+C. To the residue obtained in the test for sulfated ash add using the following expression:
1 mL of hydrochloric add R and evaporate on a water-bath.
Take up the residue in 20 mL of waler R. The solution gives 10)( 1.29 x a
reaction (a) of sodium (2.3.1).+ (100- b)m
TESTS 1.29 factor converting glycolic acid 10 sodium glycolate;
pH (2.2.3) b loss on drying as a percentage;
5.0 to 7.0 for the suspension. m mass of me subseance [0 be examined, in grams.
Shake I g with 100 mL of carbon dioxide-free waterR for

5 min. Water-soluble substances
+Sodium chloride and sodium glycolate
Disperse 10.00 g in 800.0 mL of waterR and stir for 1 min
Maximum 0.5 per cenr (dried substance) for the sum of the
every 10 min during the first 30 min. Allow to stand for I h
percentage contents.
and centrifuge if necessary. Decant 200.0 mL of the
supernatant liquid onto a fast filter paper in a vacuum
I This monograph has undergone phann~qpceial harmonisation.
See chaptt:r 5.8 PhamraaJPOeial harmonisation. filtration funnel, apply vacuum and collect 150.0 mL of the
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2022 Crospovidone 1-693
filtrate. Evaporate to dryness and dry the residue at (162+58A)C

100-105 °C for 4 h. t (7102 - SOC)
The degree of substitution is the sum of A and S.
M.aximum 10.0 per cent, determined on 1.000 g by drying in
an oven at 105 °C for 6 h. Particle size distribution (2.9.31 or 2.9.38)
Sulfated ash (2.4.14) Hausner ratio (2.9.36)
_ _- - PIIEII
14.0 per cent to 28.0 per cent (dried substance), determined
on 1.0 g.
tMicrobial contamination
TAt\1C: acceptance criterion 103 CFU/g (2.6.12).
TYMC: acceptance criterion 10 2 CFU/g (2.6.12).
Crospovidone1
Absence of EscherUhi4 coli (2.6.13).• (ph. Eur. monograph 0892)
FUNCTIONAUTY-RELATED CHARACTERISTICS
This section provides information on characteristics that are
recognised as being releuant centro! parameters for one or more
functions oj the substance when usedas an excipient (see chapter
5.15). Some of the choraaeristia described it. the Functionality-
related charaaenstics section may also be present in the mandatory
part of the monograph since they also represent mandatory quality
criteria. In such cases, a cross-reference to the tests desaibed in the M r (111.1).. 9003-39-8
mandatory pan is included in the Functionality-related
characteristics section. Control of the characteristics can contn'bute Action and use
to the quality of a medicinal product by improving the consistency Excipient in pharmaceutical products.
oj the manufacturing process and the perfonnance of the medicinal 1fIElr _
product during use. U7here control methods arecited, they are
recognised as being suitable for the purpose, but othermethods can DEFINITION
also be used. Wherever results for a particular characteristic are Cross-linked homopolymer of l-ethenylpyrrolidin-2-one.
reported, the control methodmust be indicated. Content
Thefollowing charaaeristia may be relevant jor croscarmellose 11.0 per cent to 12.8 per cent of N (Ar 14.01) (dried
sodium used as disintegratll. substance).
Settling volume 2 types of crospovidone are available, depending on the
Place 75 mL of water R in a 100 mL graduated cylinder and particle size: type A and type B.
add 1.5 g of the substance to be examined in 0.5 g portions, tCHARACTERS
shaking vigorously after each addition. Dilute to 100.0 mL
Appearance
with waler R and shake again until the substance is
Hygroscopic, white or yellowish-white powder or flakes.
homogeneously distributed. Allow to stand for 4 h.
The settling volume is between 10.0 mL and 30.0 mL. Solubility
Practically insoluble in water, in ethanol 96 per cent and in
Degree of substitution
methylene chloride.•
0.60 to 0.85 (dried substance).
Place 1.000 g in a 500 mL conical flask, add 300 mL of a IDENTIFICATION
100 gIL solution of sodium chloride Rand 25.0 mL of 0.1 M .A. Infrared absorption spectrophotometry (2.2.24).
sodium hydroxide, stopper the flask and allow to stand for Comparison crospooidone CRS.•
5 min, shaking occasionally. Add 0.25 mL of m-cresol purple B. Suspend 1 g in 10 mL of water R, add 0.1 mL of 0.05 M
solution R and about 15 mL of 0.1 M hydrochloric acid from a iodine and shake for 30 s, Add 1 mL of stardl solution Rand
burette. Insert the stopper and shake. If the solution is violet, shake. No blue colour develops within 30 s.
add 0.1 M hydrochloric add in 1 mL portions until the C. To 10 mL of waterR, add 0.1 g and shake. A suspension
solution becomes yellow, shaking after each addition. Titrate is formed and no clear solution is obtained within 15 min.
with 0.1 M sodium hydroxide until the colour turns to violet.
D. The analytical sieves must be clean and dry. For thispurpose
Calculate the number of milliequivalents (M) of base the sieves are washed in hot waterand allowed to dry O'IJeI7light in
required to neutralise the equivalent of 1 g of dried a drying cabinet at 105 °C.
substance.
Place 20 g (dried substance) in a 1000 mL conical flask, add
Calculate the degree of acid carboxymethyl substitution (A) 500 mL of waterR and shake the suspension for 30 min.
using the following expression: Pour the suspension through a 63 11m analytical sieve,
previously tared, and rinse the sieve with water R until the
1150M
filtrate is clear. Dl'Y the sieve and sample residue at 105°C
(7102 - 412M - 80C) for 5 h in a drying cabinet without circulating air. Cool in a
desiccator for 30 min and weigh.
C sulfated ash as a percentage.
Calculate the percentage sieving residue (fraction of sample
Calculate the degree of sodium carboxymethyl particles having a diameter of more than 63 11m), using the
substitution (S) using the following expression: following expression:
I This monograph has ulldergone. pharmacopoeial harmonisation:
See chapltr5.8 Pharma.opoeial harmonisation.
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1-694 Crospovidone 2022
Mobile phase aauminile for chromatography R, waterfor

chromatography R (10:90 VIV).
mass of the sieve and sample residue, after drying for 5 h, in Flow rate 1 mUmin.
grams;
initial mass of the sample. in grams;
mass of the sieve, in grams. Injection 50 ~L. After each injection of the lest solution,
wash the precolumn by passing the mobile phase backwards,
If the sieving residue fraction is more than 15 per cent, the at the same flow rate as applied in the test, for 30 min.
substance is classified as type A; if the sieving residue fraction System suiwbi/icy:
is less than or equal to 15 per cent, the substance is classified - resolution: minimum 2.0 between the peaks due to
as type B. impurity A and vinyl acetate in the chromatogram
TESTS obtained with reference solution (b);
Peroxides - reptalabilicy: maximum relative standard deviation of
Type A: maximum 400 ppm expressed as H20~ type B: 2.0 per cent determined on 6 injections of reference
maximum 1000 ppm expressed as H 202 . solution (a).
Suspend 2.0 g in 50 mL of water R. To 25 mL of this Calculation of percentage content:
suspension add 2 mL of titanium trichloride-sulfun', acid - for impurity A. use the concentration of impurity A in
reagent R. Allow to stand for 30 min and filter. reference solution (a).
The absorbance (2.2.25) of the filtrate, measured at 405 nm Limit:
using a mixture of 25 mL of a filtered 40 gIL suspension of - impurityA: maximum 10 ppm.
the substance to be examined and 2 mL of a 13 per cent VIV Loss on drying (2.2.32)
solution of sulfuric acid R as the compensation liquid. has a Maximum 5.0 per cent. determined on 0.500 g by drying in
maximum of 0.35. an oven at 105 "C,
For type B use 10 mL of the suspension and dilute to 25 mL Sulfated ash (2.4.14)
with water R for the test. Maximum 0.1 per cent, determined on 1.0 g.
Water-soluble substances
ASSAY
Place 0.100 g of the substance to be examined (m mg) in a
Place 25.0 g in a 400 mL beaker. add 200 mL of water R combustion flask and add 5 g of a mixture of I g of copper
and stir for 1 h using a magnetic stirrer. Transfer the sulfate pentahydrate R. 1 g of titanium dioxide Rand 33 g of
suspension to a 250.0 mL volumetric flask. rinsing with dipolaSS1'um sulfate R. and 3 glass beads. Wash any adhering
water R. and dilute to volume with the same solvent. Allow particles from the neck into the flask with a small quantity of
the bulk of the solids to settle. Filter about 100 mL of the water R. Add 7 mL of sulfuric acid R. allowing it to run down
almost clear supernatant through a membrane filter (nominal the inside wall of the flask. Gradually heat the flask until the
pore size 0.45 J,Un), protected by superimposing a membrane solution has a clear. yellowish-green colour. and the inside
filter (nominal pore size 3 um), While filtering. stir the liquid wall of the flask is free from carbonised material. and then
above the membrane filter manua1Jy or by means of a heat for a further 45 min. After cooling, cautiously add
mechanical stirrer. taking care not to damage the membrane 20 mL of water R. and connect the flask to the distillation
filter. Transfer 50.0 mL of the clear filtrate to a tared apparatus, which has been previously washed by passing
100 mL beaker. evaporate to dryness and dry at 105-110 "C steam through it. To the absorption flask add 30 mL of a
for 3 h. The residue weighs a maximum of75 mg. 40 gIL solution of boric acid R, O. I 5 mL of bromocresol green-
Impurity A methyl red soluuon R and sufficient water R to immerse the
Liquid chromatography (2.2.29). lower end of the condenser tube. Add 30 mL of strong sodium
Test solution Suspend 1.250 g in 50.0 mL ofmelhanol Rand hydroxick solution R through a funnel. cautiously rinse the
shake for 60 min. Leave the bulk to settle and filter through funnel with 10 mL of water R. immediately close the clamp
a membrane filter (nominal pore size 0.2 um), artached to the rubber tube. then start the distillation with
Reference solution (a) Dissolve 50 mg of l-vinylpyrrolidin-2- steam to obtain 80-100 mL of distillate. Remove the
one R (impurity A) in methanol R and dilute to 100.0 mL absorption flask from the lower end of the condenser tube,
with the same solvent. Dilute 1.0 mL of the solution to rinsing the end part with a small quantity of water R. and
100.0 mL with methanol R. Dilute 5.0 mL of this solution to titrate the distillate with 0.025 M sulfun"c acid until the colour
100.0 mL with the mobile phase. of the solution changes from green through pale greyish-blue
to pale greyish red-purple. Carry out a blank determination
Reference solution (b) Dissolve 10 mg of l-'Uinylpyn-olidin-2- and make any necessary correction.
on'e R (impurity A) and 0.50 g of vinyl aceuue R in
methanol R and dilute to 100 mL with the same solvent. 1 mL of 0.025 M sulfuric acid is equivalent to 0.700 mg of N.
Dilute 1 mL of the solution to 100 mL with the mobile STORAGE
phase. In an airtight container.
Precolumn: +LABELLING
- size: 1= 0.025 m, 0 ::: 4 mrn; The label states the type of crospovidone (type A or type B).
- stationary phase: end-capped oClad«y/siJy1 silitagelfor
chromatography R (5 um). •
Column:
- size: 1 = 0.25 m, 0 ::: 4 rnm;
- stationary phase: polarend-capped octaduylsilyl silica gelfor
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2022 Crotarniton 1-695
PhEIr _
IMPURITIES _~
DEFINITION
N-Ethyl-N-(2-methylphenyl)but-2-enamide.
Content
- sum of the (B)- and (Z)-isomers: 96.0 per cent to
102.0 per cent;
A. l-ethenylpyrrolidin-2-one (l-vinylpyrrolidin-2-one). - (Z)-isomer: maximum 15.0 per cent.
FUNCTIONALITY-RELATED CHARACTERISTICS CHARACTERS
This section provides infonnation on characteristics that are Appearance
recogmsed as being relevant centrol parameters for one ormore Colourless or pale yellow, oily liquid.
functions of the substance when used as at' excipient (see chapter Solubility
5.15). Some of the characteristics described in the Functionality- Slightly soluble in water, miscible with ethanol (96 per cent).
related characteristics section may also be present in the mandatory At low temperatures it may partly or completely solidify.
part of the monograph since they also represent mandatory qualil)J
criteria. In such cases, a cross-reference to the tests described in the IDENTIFICATION
mandatory part is included in the Functionality-related First identification: B.
characteristics section. Control of the characteristics can contn'bute Second identlfication: A, C, D.
to the qualityof a medicinal product by improving the consistency A. Ultraviolet and visible absorption spectrophotometry
of the mamifactun'ng process and the peifonnance of the medicinal (2.2.25).
product during use. Where control methods are cited, they are Test solution Dissolve 25.0 mg in cyclohexane R and dilute to
recognised as being suitable for the purpose, but othermethods can
also be used. Wherever restdts for a particular characteristic are solution to 10.0 mL with cydohexane R.
reported, the control methodmust be indica led.
Spectralrange 220-300 run.
The following characteristics may be relevant for crospovidone used
as disimegrant.
Absorption maximum At 242 om.
Hydration capacity
Introduce 2.0 g into a 100 mL centrifuge tube and add B. Infrared absorption spectrophotometry (2.2.24).
40 mL of water R. Shake vigorously until a suspension is Comparison crotamiton CRS.
obtained. Shake again 5 min and 10 min later, then C. Thin-layer chromatography (2.2.27).
centrifuge for 15 min at 750 g. Decant the supernatant and Test solution Dissolve 25 mg of the substance to be
weigh the residue. The hydration capacity is the ratio of the examined in anhydrous ethanol R and dilute to 10 mL with
mass of the residue to the initial mass of the sample. It is the same solvent.
typically 3 to 9.
Reference solution Dissolve 25 mg of crotamiton CRS in
Particle-size distribution (2.9.31) anhydrous ethanolR and dilute to 10 mL with me same
Powder flow (2.9.36) solvent.
The following characteristic may be relevant for crospcvidone used Plate TLC silica gel F25 4 plate R.
as suspension swbiliser. Mobilephase Shake 98 volumes of methylene chloride R with
Settling volume 2 volumes of concentrated ammonia R, dry over anhydrous
Introduce 10 g into a 100 mL graduated cylinder and add sodium sulfate R, filter and mix 97 volumes of the filtrate with
90 mL of water R. Shake vigorously. Dilute to 100 mL with 3 volumes of 2-propanol R.
waterR, washing me powder residues from me walls of the Application 5 j.lL.
cylinder. Allow to stand for 24 h, men read me volume of
Development Over a 2/3 of the plate.
the sediment. It is typically greater than 60 mL.
________________ ~~~_ PlJEIJ1
Drying In air.
Crotamiton reference solution.
(ph. Bur. monograph 1194) D. To 10 mL of a saturated solution add a few drops of a
3 gIL solution of potassium permanganate R. A brown colour
is produced and a brown precipitate is formed on standing.
and (Z}-isomer TESTS
1.006 to 1.011.
203.3 483-63-6 1.540 to 1.542.
Action and use Free amines

Acaricide. Maximum 500 ppm, expressed as ethylaminotoluene.
Dissolve 5.00 g in 16 mL of methylene chloride R and add
Preparations
4.0 mL of glacial acetic acid R. Add 0.1 mL of metani/yellow
Crotamiton Cream
Crotamiton Lotion
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1-696 Cyanocobalamin 2022
soludon R and 1.0 mL of 0.02 M perch/oric acid. The solution the chromatogram obtained with reference solution (c)
is red-violet. (1.0 per cent);
Chlorides - disregard limit: 0.02 times the sum of the areas of the
Maximum 100 ppm. peaks due to the (Z)- and (E)-isomers in the
Boil 5.0 g under a reflux condenser for 1 h with 25 mL of
(0.02 per cent).
ethanol (96 per cent) Rand 5 mL of a 200 gIL solution of
sodium hydroxide R. Cool, add 5 mL of uuuer R and shake Sulfated ash (2.4.14)
with 25 mL of ether R. Dilute the lower layer to 20 mL with Maximum 0.1 per cent, determined on 1.0 g.
waterR; add 5 mL of nitric add R, dilute to 50 mL with ASSAY
water R and add 1 mL of a freshly prepared 50 gIL solution Liquid chromatography (2.2.29) as described in the test for
of silver nitrate R. Any opalescence in the solution is not more related substances with the following modification.
intense than that in a mixture of 1 mL of a freshly prepared
50 gIL solution of silver nitrate R and a solution prepared by
diluting 5 mL of a 200 gIL solution of sodium hydroxide R to Calculate the percentage content of C 13 H I 7NO from the sum
20 mL with water R and adding 1.5 mL of 0.01 M . of the areas of the peaks due to the (Z)- and (E)-isomers in
hydrochloric acid, 5 mL of nitric add R and diluting to 50 mL the chromatograms obtained. Calculate the content of the
with water R. (Z)-isomer, as a percentage of the total content of the (E)-
and (Z)-isomers. from the chromatogram obtained with test
Related substances solution (b).
Testsolution (a) Dissolve 50,0 rngof the substance to be STORAGE
examined in the mobile phase and dilute to 100.0 mL with Protected from light.
Testsolution (b) Dilute 1.0 mL of test solution (a) to Specified impuniies A.
Referena solution (a) Dissolve 50.0 mg of crotamuon CRS in
phase. Dilute 1.0 mL of this solution to 20.0 mL with the
mobile phase.
Re/erena sdution (b) Dissolve 15.0 mg of crotamiton
impun'ty A CRS in the mobile phase and dilute to 20.0 mL A. N-ethyl-N-(2-methylphenyl)but-3-enamide.
with the mobile phase. Dilute 1.0 mL of this solution to ___________________ ~ PhEll
Referenc« solution (c) Dilute 1.0 mL of test solution (a) to
Referena solution (d) Dissolve 15 mg of crotamium Cyanocobalamin ***
impun'ty A CRS in the mobile phase and dilute to 100.0 mL ** **
with the mobile phase. Dilute 1.0 mL of this solution to (ph. Bur. monograph 0547) **.**
10.0 mL with test solution (a).
Column:
- size: I;;;;; 0.25 m, 0 ;;;;; 4 mrn;
- stationary phase: silica gelfor chromatography R (5 urn),
Mobile phase tetrahydrojuratl R, cydohexane R (8:92 VIV).
Flow rate 1.0 mLlmin.
Inject;on 20 J.lL of test solution (a) and reference
Run time 2.5 times the retention time of the (E)-isomer.
Relative retention With reference to the (E)-isomer:
(Z)-isomer ;;;;; about 0.5; impurity A ;;;;; about 0.8.
System suiUJbiJity Reference solution (d): ';
impurity A and the (E)-isomer.
Limus:
- impun'ty A: not more than the area of the corresponding
- unspecified impurities: for each impurity, not more than 1355 68-19-9
0.1 times the sum of the areas of the peaks due to the
Action and use
(Z)- and (E)- isomers in the chromatogram obtained with
Vitamin B 12 analogue.
- sum of impurities other than A: not more than the sum of Preparations
the areas of the peaks due to the (Z)- and (E)-isomers in Cyanocobalamin Oral Solution
Cyanocobalamin Tablets
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2022 Cyanocobalamin 1-697
PhE<I ~--_------- Time Moblle phase A Moblle phase B

(min) (per cent VIJI) {per cent V/Vl
DEFINITION 0- I 90 10
Crxt-[~-(5,6-Dimethy1benzimidazo1yl)I-Coll_cyanocobamide. I • 16 90 --+ 80 JO -lo 20
Content 16 - 23 80 --+ 60 20 --+ 40
This monograph applies to cyanocobalamin produced by Flow rate 0.4 mUmin.
fermentation. Detection Spectrophotometer at 361 nm.
CHARACTERS Injedon 3 ~L.
Appearance Identification of impurities Use the chromatogram supplied .
Dark red, crystalline powder or dark red crystals. with cyanocobalamin for system suitability CRS and the
Solubility chromatogram obtained with reference solution (b) to
Sparingly soluble in water and in ethanol (96 per cent), identify me peaks due (Q impurities A, C, E and F; use me
practically insoluble in acetone. chromatogram supplied with cyanocobalamin for peak
The anhydrous substance is very hygroscopic. identification CRS and the chromatogram obtained with
reference solution (d) to identify the peaks due to
IDENTIFICATION impurities Band D.
A. Ultraviolet and visible absorption spectrophotometry Relativeretention WiLh reference to cyanocobalamin
(2.2.25).
=
(retention time about 10 min): impurity F about 1.06;=
Test solution Dissolve 2.5 mg in waterR and dilute to impurity D = about 1.18j impurity C = about 1.23;
100.0 mL with the same solvent. =
impurity A about 1.26; impurity E about 1.33; =
Spectral range 260-610 om. impurity B = about 1.45.
Absorption maxima 278 om, 361 om and 547-559 nm. System suitabuity Reference solution (b):
Absorbance raWs: - resolution: minimum 1.4 between the peaks due to
- A:J61 I A547.S59 = 3.15 to 3.45; impurities C and A;
- A'61 I A27S = 1.70 to 1.90. - peak-co-'Valley ratio: minimum 2.5, where H p = height
B. Examine the chromatograms obtained in the test for above the baseline of the peak due to impurity F and
Hv = height above me baseline of the lowest point of me
related substances.
Results The principal peak in the chromatogram obtained cyanocobalamin.
with the test solution is.similarin retention time and size to
the principal peak in the chromatogram obtained with Limits:
reference solution (c). - impuniy C: maximum 1.5 per cent;
- impwlty A: maximum 0.7 per cent;
TESTS - impurities B;, D, EJ F: for each impurity, maximum
Related substances 0.5 per centj
Liquid chromatography (2.2.29): use the normalisation - unspecified impun·ties: for each impurity, maximum
procedure. Store the solutions at 2-8 °c;, protected from light;, 0.2 per cent;
and use them within 24 h. - total: maximwn 3.0 per cent;
Test solution Dissolve 25.0 mg of the substance to be - reporting threshold: 0.10 per cent (reference solution (ej).
examined in water R and dilute to 50.0 mL with the same The thresholds indicated under Related substances
solvent. (Table 2034.-1) in the general monograph Substancesfor
Reference solutWn (a) Dilute 1.0 mL of the test solution to pharmaceutical use (2034) do not apply.
100.0 mL with water R. Dilute 1.0 mL of this solution to Loss on drying (2.2.32)
10.0 mL with water R. Maximwn 12.0 per cent, determined on 0.400 g by drying in
Reference solutWn (b) Dissolve 5 mg of cyanrxobalamin for vacuo at 105°C for 2 h.
system suitability CRS (containing impurities A, C, E and F) ASSAY
in water R and dilute to 10 mL with the same solvent.
Reference solution (c) Dissolve 5.0 mg of same solvent, Dilute 25.0 mL of the solution to 200.0 mL
cyanocobalamin CRS in water R and dilute to 10.0 mL with with water R. Measure the absorbance (2.2.25) at the
the same solvent. absorption maximwn at 361 run.
Reference solutWn (d) Dissolve 5 mg of cyanocobalamin for Calculate the content of C63HssCoN14014P taking the
peak identification CRS (containing impurities B and D) in specific absorbance to be 207.
water R and dilute £0 10 mL with the same solvent.
STORAGE
Column:
- size: 1= 0.15 IJl, {£) = 2.1 mm;
- stationary phase: end-eapped elhylene-bridged rxty/silyl silica IMPURITIES
gelfor chromatography (hybridmaurial) R (I. 7 pm): Specified impuniies A, B, CJ D, E, F.
- temperature: 60°C. Otherdetectable impurities (the following substances would;, if
Mobue phase: present at a sufficientlevel be detected by one or otherof the tests
J
- mobile phase A: 1.0 gIL solution of ammonium formate R in the monograph. They are limited by the general acceptance
adjusted to pH 3.5 with anhydrous formic acid R; criterion for other/unspecified impurities. It is therefore not
- mobile phase E: methanclR; necessary to idemify these impurities for demonstration of
pharmaceutical use) G, H.
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1-698 Cyanocobalamin 2022
A. Ct:n.-[a:-(5,6-dimethylbenzimidazolyl))-Co~-cyano-8 D. Coa-[a:-(S,6-dimethylbenzimidazolyl)]-Co~-cyanocobamic
hydroxycobarnic o,b,d,e,g-pentaamide ,,8-lactone a,c,d,e,g-pentaamide (cyanocobalamin b-monocarboxylic
(cyanocobalamin e,8-lactone, cyanocobalamin 7~,8~ acid. 32-carboxycyanocobalamin)
lactone),
E. (BR)-Coa-[Ct-(S,6-dimethylbenzimidazolyl)]-CoP-
B. CtXJ.-[a:-(S,6-dimethylbenzimidazolyl»)-Cop-cyanocobamic
cyanocobamide (8-epi-cyanocobalamin»
a,b,e,d,g-pentaamide (cyanocobalamin e-rnonocarboxylic
acid, SO-carboxycyanocobalamin) F. unknown structure (cyanocobalamin isomer).
o H2N o ~
HN----f H2N----f
C~ \......... H CH:\
He!
3
\.......... H
OHC '
a .... "'"
.'
CH
3~
0
°HC \
3 ....
... NH
Hi'! CN H 2
N , N_
'C::
/,",
N : N
f ~ CHa
~N ~ C~
(~)"'CHa! C~ H "r:
H3C~f-? 0 #~--(YCHa
O;;p~CH'
HO
C. Coa-[a:-(5,6-dimethylbenzirnidazolyl)]-Cop-cyano-b-N- G. Coa:-[a:-(5,6-dimethylbenzimidazolyl)]-Cop-cyano-e-N-
methylcobamide (b-N-methylcyanocobalamin, methylcobamide (e-N-methylcyanocobaJamin~
34-methylcyanocobalamin) 50-methylcyanocobalamin),
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2022 Cyclizine 1-699
Chloride
Dissolve 0.20 g in 2 mL of methanol and dilute to 30 mL
with 2M nitric acid. 15 mL of the resulting solution complies
with the limit test for chlorides, Appendix VII (500 ppm).
Related substances
Carry out the method for gas chromawgraphy,
Appendix 1lI B, using the following solutions in methanol
prepared immediately before use.
(1) 0.5% w/v of the substance being examined.
(2) Dilute I volume of solution (1) to 100 volumes and
further dilute 1 volume of the resulting solution to
10 volumes.
(3) 0.0025% wlv of cydieine hydrochloride BPCRS,
0.0025% wlv of I-methy/piperazine BPCRS (impurity A) and
0.0025% wlv of diphenylmelhanol BPCRS (impurity B).
(a) Use a fused silica column (25 m x 0.33 mm) coated
with a 0.5-J.lm film of phenyl(5)merhyl(95)polysiloxane (HP-5
H. C()(J.-[o:-(5,6-dimethylbenzimidazolyl)]-Cojl- is suitable).
hydroxocobamide (hydroxocobalamin).
(b) Use helium as the carrier gas at a flow rate of 1 mL per
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Ph€/I
minute.
(c) Use the gradient conditions described below.
(d) Use a split injection ratio of 1:25.
(e) Use a flame ionisation detector at 290°.
Cyclizine
(f) Inject 1 J.lLof each solution.
(g) The peaks elute in the order: methanol,
l-methylpiperazine, diphenylmethanol, cyclizine.
Time Temperature Comment

(minutes)
O--?14 100° --?240· linear gradient

266.4 82-92-8 14-?16 240° --?270° linear gradient
Action and use 16--?30 270· isocratic
Histamine Hi receptor antagonist; antihistamine.
Preparation
Cyclizine Injection SYSTEM SUITABILITY
Inject solution (3) 6 times. The relative standard deviation of
DEFINITION each of the areas of the 3 principal peaks is not more than
Cyclizine is l-benzhydryl-d-methylpiperazine, It contains not 5.0%.
less than 98.5% and not more than 101.0% of ClsH22N2, The test is not valid unless, in the chromatogram obtained
calculated with reference to the dried substance. with solution (3);
CHARACTERISTICS the peak-to-1Jalley ratio between methanol and
A white or creamy white, crystalline powder. l-methylplperazine (impurity A) is at least 50;
Practically insoluble in water. It dissolves in most organic the resolutwn factor between diphenylmethanol (impurity B)
solvents and in dilute acids. and cyclizine is at least 18.
IDENTIFICATION LIMITS
A. The infrared absorption spectrum, Appendix II A, is In the chromatogram obtained with solution (1):
concordant with the reference spectrum of cyclizine (RS 075). the area of the peak corresponding to l-methylpiperazine
B. Melting poim, about 1070, Appendix V A. (impurity A) is not greater than the peak corresponding to
l-methylpiperazine in solution (3) (0.5%)j
TESTS
Alkalinity the area of the peak corresponding to diphenylmethanol
Shake 1 g with 25 mL of carbon dioxide-free water for (inJpurity B) is not greater than the peak corresponding to
5 minutes and filter. The pH of the filtrate is 7.6 to 8.6, diphenylmetbanol in solution (3) (O.5%)j
Appendix V L. the area of any other secondary peak is not greater than the
Clarity of solution area of the principal peak in the chromatogram obtained with
A 1.0% wlv solution in ether and a 1.0% wlv solution in solution (2) (0.1%);
2M hydrochloric acid are clear, Appendix IV A. the sum of the areas of aU secondary peaks is not greater than
10 times the area of the principal peak in the chromatogram
obtained with solution (2) (1.0%).
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1-700 Cyclizine Hydrochloride 2022
Disregard any peak with an area less than 0.5 times that of B. Infrared absorption spectrophotometry (2.2.24).
the principal peak in the chromatogram obtained with Comparison cydizine h)'Cirochloride CRS.
solution (2) (0.05%).
Loss on drying Testsolution Dissolve 10 mg of me substance to be
Q
When dried to constant weight at 80 , loses not more than examined in methanol R and dilute to 10 mL with me same
1.0% of its weight. Use 1 g. solvent.
Sulfated ash Reference solution Dissolve 10 mg of cydizine
Not more than 0.1 %, Appendix IX A. hydrochloride CRS in methanol R and dilute to 10 mL with the
ASSAY same solvent.
Carry out Method I for non-aqueous tierauOn. Piau TLC silica gel GF254 piau R.
Appendix VIII A, using 0.1 g and determining the end point Mobile phase concentnued ammonia R, methanol R. methylene
potentiometrically. Each mL of 0.1M perchloric acid VS is chloride R (2: 13:85 VIVIJI).
equivalent to 13.32 mg of C u Jl nN2 • Application 20 ~L.
Cyclizine Hydrochloride *** Deuction Expose to iodine vapour for 10 min.
*** *** Results The principal spot in the chromatogram obtained
(ph. Bur. monograph 1092) *** with the test solution is similar in position, colour and size to
reference solution.
D. Dissolve 0.5 gin 10 mL of ethanol (60 percent V/~ R,
• Hel heating if necessary. Cool in iced water. Add 1 mL of diluu
sodium hydroxide so/utUm Rand 10 mL of waUl' R. Filter,
wash the precipitate with uuue» R and dry at 60°C at a
pressure not exceeding 0.7 kPa for 2 h. The melting point
(2.2.14) is 105 QC to 108 QC.
302.8 303-25-3 E. It gives reaction (a) of chlorides (2.3.1).
Histamine HI receptor antagonist, antihistamine. pH (2.2.3)
Preparation 4.5 to 5.5.
Cyclizine Tablets Dissolve 0.5 g in a mixture of 40 volumes of ethanol
(96 per cent) R and 60 volumes of carbon dibxide-jree waterR
PIIEII _
and dilute to 25 mL with the same mixture of solvents.
1-(Diphenylmethyl)-4-methylpiperazine hydrochloride. Gas chromatography (2.2.28). Prepare the solutions immediately
Content before use.
98.5 per cent to 101.0 per cent (dried substance). Test sdution Dissolve 0.250 g of the substance to be
examined in 4.0 mL of medianol R and dilute to 5.0 mL with
CHARACTERS
1 M sodium hydroxide.
Appearance
White or almost white, crystalline powder. Reference solution (a) Dissolve 25 mg of the substance to be
examined in 10.0 mL of methanol R. Dilute 1.0 mL of this
Solubility solution to 50.0 mL with methatwl R.
IDENfIFICATION examined, 5.0 mg of cydizine impurity A CRS and 5.0 mg of
Piru identification: BJ E. cyclizine impurity B CRS in methanol R and dilute to 20.0 mL
Second identification: A, ,CJ D, B. with the same solvent.
(2.2.25). - material: fused silica;
Test solution (a) Dissolve 20.0 mg in a 5 gIL solution of = =
- size: I 25 m, 0 0.33 mm;
sulftmc acid R and dilute to 100.0 mL with the same acid - sratronary phase: phenyl(5)methyl(95)poiysiloxane R (film
solution. thickness 0.50 urn),
Test solution (b) Dilute 10.0 mL of test solution (a) to Camer gas helium for chromatography R.
100.0 mL with a 5 gIL solution of sulfuric acid R. Flow rate 1.0 mIlmin.
Spectral range 240-350 run for test solution (a); 210-240 run Split ratio 1:25.
for test solution (b).
Resolution (2.2.25): minimum 1.7.
Absorption maxima At 258 run and 262 run for test
solution (a); at 225 nm for test solution (b).
Absorbance rauo A26?!A258 = 1.0 to 1.1.
Specific absorbance at the absorption maximum at 225 nm
370 to 410 for test solution (b).
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2022 Cyclopenthiazide 1-701
Temperature:
Cyclopenthiazide
Tbn. Temperature
(min) Cq o 0 0 0
*/1 \\1/
Column 0- 14
J4·16
100 -+ 240
240 -e 270
H2 N , S Y y S ' N H r-,
Injectionport
16 - 30 270
250
CI~N~
H
Detector 290
379.9 742-21J-1
Injection I ~L. Action and use
Relative retention With reference to cyclizine (retention Thiazide-diuretic.
impurity B = about 0.7. DEFINITION
System suitabt7ity Reference solution (b): Cyclopenthiazide is 6-chloro-3-eyclopentylmethyl-3,4-
- peak-to-vaUey ratio: minimum 50, where Hp = height dihydro-I, 2,4-benzothiadiazine-7-sulfonamide I, I-dioxide.
above the baseline of the peak due to impurity A and It contains not less than 98.0% and not more than 102.0%
HI.> = height above the baseline of the lowest point of the of C13HlSCINJ04S2, calculated with reference to the dried
curve separating this peak from the peak due to methanol. substance.
Limits: CHARACTERISTICS
- impurities A, B: for each impurity, not more than the area A white powder.
of the corresponding peak in the chromatogram obtained PracticaUy insoluble in water; soluble in acetone and in erhanol
with reference solution (b) (0.5 per cent); (96%); very slightly soluble in ether.
area of the principal peak ln. the chromatogram obtained IDENTIFICATION
with reference solution (a) (0.10 per cent); A. The infrared absorption spectrum, Appendix II A, is
- total: not more than 10 times the area of the principal concordant with the reference. spectrum of cyclopenthiazide
peak in the chromatogram obtained with reference (RS 077).
solution (a) (1.0 per cent); B. The light absorption, Appendix II B, in the range 230 10
- disregard limit: 0.5 times the area of the principal peak in 350 run of a 0.002% wlv solution in O.OIM sodium hydroxide
the chromatogram obtainedwith reference solution (a) exhibits two maxima, at 273 run and 320 run. The absorbance
(0.05 per cent). at 273 run is about 0.88 and at 320 run is about 0.12.
Loss on drying (2.2.32) C. Carry OUI the method for thin-I~ chromatography,
Maximum 1.0 per cent, determined on 1.000 g by drying in Appendix ill A, using silica gel GF2 '54 as the coating
an oven at 130°C. substance and ethylacetate as the mobilephase. Apply
Sulfated ash (2.4.14) separately to the plate 5 ul, of each of two solutions in
Maximwn 0.1 per cent, determined on 1.0 g. acetone containing (1) 0.1 % wlv of the substance being
examined and (2) 0.1% wlv of eycIopenthiazide BPCRS. After
ASSAY removal of the plate, dry it in a current of air, examine under
In orderto awid overheating in the reaction medium, mix ultraviolet light (254 nm) and then reveal the SPOlS by
thoroughly throughoul and stopthe titration immediately after'he Method I. By each method of visualisation the principal spot
end-point has been reached. in the chromatogram obtainedwith solution (1) corresponds
Dissolve 0.120 g in IS mL of anhydrous formic acid Rand in colour and intensity to mat in the chromatogram obtained
add 40 mL of acetic anhydride R. Titrate with 0.1 M perchleric with solution (2).
acid, determining the end-pointpotentiometrically (2.2.2(/).
TESTS
I mL of 0.1 M perchlmic acid is equivalent 10 15.14 mg Related substances
of CI!~H2JCIN2' Carry out the method for thin-layer chromatagraphy,
STORAGE Appendix ill A, using silic.a gel G as the coating substance
Protected from light. and ethyl acetate as the mobile phase. Apply separately to the
IMPURITIES plate 5 IlL of each of two solutions of the substance being
Specified ;mpun'lies A, B. examined in acetane containing (I) 0.50% wlv and (2)
0.0050% wlv. Afterremoval of the plate) dryit in a current
of air and reveal the spots by Method 1. Any secondory spo' in
the chromatogram obtainedwith solution (1) is not more
A. l-methylpiperazine, solution (2).
Loss on drying
When dried to constant weight at 105°, loses not mere than
Sulfated ash
Not more than 0.1 %, Appendix IX. A.
ASSAY
B. diphenylmethanol (benzhydrcl). Dissolve 0.5 g in 50 mL of butylamine and carry out
________ ~ PhE" Method IT for non-aqueous tiaauon, Appendix VIII A, using
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1-702 Cyclopentolate Hydrochloride 2022
a.1M.. tetrabutylammonium hydroxide VS as titrant and Delation Spray with akoholic solution of sulfuric acid Rand
magn~san solutionas indicator; titrate to a pure blue end heat at J20 °C for 30 min; examine in ultraviolet light at
point. Each mL of O.1M tetrabutylammonium hydroxide VS is 365 om.
equivalent to 18.99 mg OfCI3HISClNJ04S2. Result The principal spot in the chromatogram obtained
with the test solution is similar in position, fluorescence and
size to the principal spot in the chromatogram obtained with
the reference solution.
Cyclopentolate Hydrochloride ***
*** *** D. It gives reaction (a) of chlorides (2.3.1).
(ph. Eur. monograph 1093) *** TESTS
pH (2.2.3)
4.5 to 5.5.
and enantiomer , HCI 20 mL with the same solvent.
Related substances
Liquid chromatography (2.2.29). Prepare the sohuions
327.8 5870-29-1 Test solution Dissolve 20 mg of the substance to be
Action and use solvent.
Anticholinergic.
Preparation 100.0 mL with water R. Dilute 5.0 mL of this solution to
Cyclopentolare Eye Drops 10.0 mL with water R.
PhE" ~ _ Reference solulian (b) Dissolve 10 mg of cydopemokue for
system suitabililjl CRS (containing impurity C) in water Rand
DEFINITION dilute to io.o mL with the same solvent.
2-(Dimethylamino)ethyl (2R5)-(I-
Column:
hydroxycyclopentyl)(phenyl)acetate hydrochloride.
- size: / =: 0.125 m, 0 =: 4.0 nun;
Content - stationary phase: spherical end-<:apped hexylsilyl Sl7lea gelfor
98.5 per cent to IOI.5 per cent (dried substance). chromatography R (5 1'1II).
CHARACTERS Mobile phase Dissolve 0.66 g of ammoniumphosphate R in
Appearance water R, adjust to pH 3.0 with phosphoric acid R and dilute to
White or almost white, crystalline powder. 1000 mL with water R; mix and filter; mix 55 volumes of this
Solubility solution and 45 volumes of acetonitrile RI.
Very soluble in water, freely soluble in ethanol (96 per cent). Flow rare 1.0 mUmin.
It shows polymorphism (5.9). Detection Spectrophotometer at 220 run.
IDENTIFICATION lnje<rion 20~.
First identification: B, D. Run time 2.5 times the retention time of cyclopentolate.
Second identification: A, C, D. JdentijicatUm of impun'ties Use the chromatogram supplied
with cycIcpentoiate for system suitabililjl CRS and the
A. Melting point (2.2.14): 135°C to 141°C.
B. Infrared absorption spectrophotometry (2.2.24). identify the peak due 10 impurity C.
Preparation Discs of potassium chloride R. Relative retention With reference to cyclopentolate (retention
Comporison cyclopenwlote hydrochloride CRS. =
time about 4 min): impurity C about 0.9.=
If me spectra obtained show differences, dissolve the System suitability Reference solution (b):
substance to be examined and the reference substance - peak-to-valley ratio: minimwn 6, where Hp =: height above
separately in ethanol (96 per cent) R, evaporate to dryness and the baseline-of the peak due to impurity C and
record new spectra using the residues. HI} =: height above the baseline of the lowest point of the
C. Thin-layer chromatography (2.2.27). curve separating this peak from the peak due to
Test solution Dissolve 10 mg of the substance to be cyclopentolate.
examined in 5 mL of ethanol (96 per cent) R. Limits:
Reference solution Dissolve 10 mg of cydqpentolate - correction factor: for the calculation of content, multiply the
hydrochloride CRS in ethanol (96 per cenO R and dilute to peak area of impurity C by 2.0;
5 mL with the same solvent. - impun'ty C: not more than me area of the principal peak
(0.5 per cent);
Alobile phase concentrated ammonia R, waterR, butyl - unspecified impurities: for each impurity, not more than
acetate R, 2-propatlol R (5:15:30:50 VIVIV/V). 0.2 times the area of the principal peak in the .
Appliwtian 10 ~L. chromatogram obtained with reference solution (a)
Development Over 2/3 of the plate. (0.10 per cent);
Drying In air. - total: not more than twice the area of the principal peak in
(1.0 per cent);
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2022 Cyclophosphamide 1-703
- disregard limit: 0.1 times the area of the principal peak in PhE" _
the chromatogram obtained with reference solution (a) DEFINITION
(0.05 per cent).
Cyclophosphamide contains not less than 98.0 per cent and
Loss on drying (2.2.32) not more than the equivalent of 102.0 per cent of (2RS)-N,
Maximum 0.5 per cent, determined on 1.000 g by drying in N-bis(2-chloroethyl)tetrahydro-2H-I,3,2-oxazaphosphorin-2-
an oven at lOS °C for 4 h. amine z-oxide, calculated with reference to the anhydrous
Sulfated ash (2.4.14) substance.
Maximum 0.1 per cent, determined on 1.0 g. CHARACTERS
ASSAY A white or almost white, crystalline powder, soluble in water,
Dissolve 0.250 g in a mixture of 1.0 mL of 0.1 M hydTOChlori< freely soluble in alcohol.
acid and 50 mL of ethanol (96 per cen!! R. Carry out a IDENTIFICATION
potentiometric titration (2.2.20)J using 0.1 M sodium First identification: B.
inflexion.
A. Determine the meltingpoint (2.2.14) of the substance to
be examined. .Mix equal pans of the substance to be
C17H26CIN03- examined and cyclophosphamide CRS and determine the
IMPURITIES melting point of the mixture. The difference between the
Specified impuniles C. melting points (which are about 51 °C) is not greater than
Otherdet«eable impurities (the following substances would, if 2°C.
present at a sufficient level, be detected by one or other of the tests B. Examine by infrared absorption spectrophotometry
in the monograph. They are limiud by thegeneral acceptance (2.2.24), comparing with the spectrum obtained with
cruetion for other/unspecified impurities and/or by thegeneral cyclophosphamide CRS.
monograph Subsran", for pharmaceutical use (2034). It is C. Examine the chromatograms obtained in the test for
therefore not neussary W identify these impurities for related substances. The principal spot in the chromatogram
demonstration of compliance. See also 5.10. Control of impurities obtainedwith test solution (b) is similar in position, colour
in substances for pharmaceutical we) A, B. and size to the principal spot in the chromatogram obtained
OvCo,H
HOOH andenentcmer
D. Dissolve 0.1 gin 10 mL of water R and add 5 mL of
silvernitrate solution Rt, the solution remains clear. Boil, a
white precipitate is formed whichdissolves in concentrated
ammonia R and is reprecipitated on the addition of dilute
nitne acid R.
A. (2RS)-(I-hydroxycyclopentyl)(phenyl)acetic acid,
B. phenylacetic acid,
o; TESTS
Solution S
('I yH3 9 than reference solution Y. (2.2.2, Method11).
V00~N"'CH3 pH (2.2.3)
The pH of solution S is 4.0 to 6.0, determined immediately
C. 2-(dimethylamino)ethyl phenylacetate.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
after preparation of the solution.
Related substances
Examine by thin-layer chromatography (2.2.27)) using silica
gel GRas the coating substance.
Cyclophosphamide *** Test solution (a) Dissolve 0.10 g of the substance to be
*** *** examinedin akohol R and dilute to 5 mL with the same
(Ph. Eur. monograph 0711) *** solvent.
Tess solution (b) Dilute I mL of test solution (a) to 10 mL
with alcohd R.
• H,O Reference solution (a) Dissolve 10 mg of
cyclophosphamide CRS in alcohol R and dilute to 5 mL with
the same solvent.
Reference solmion (b) Dilute 0.1 mL of test solution (a) (0
C,H15Cl,N,O,P,H2 0 279.1 6055-19-2 10 mL with akohol R.
Apply separately to the plate 10 JAL of each solution: Develop
Action and use
over a path of 15 em usinga mixture of 2 volumes of
Cytotoxicalkylating agent.
anhydrous formic add R, 4 volumes of aceume RJ 12 volumes
Preparations of waterR and 80 volumes of methyl ethylketone R. Dry the
Cyclophosphamide Injection plate in a current of wann airand heat at 110 °C for 10 min.
Cyclophosphamide Oral Solution At the bottom of a chromatographic tank, place an
Cyclophosphamide Tablets evaporating dish containing a 50 gIL solution of potassium
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1-704 Cyproheptadine Hydrochloride 2022
permanganate R and add an equalvolumeof hydrochloric Content

acid R. Place the plate whilst still hot in the tank and close 98.5 per cent to 101.0 per cent (anhydrous substance).
the tank. Leave the plate in contactwith the chlorine gas for
CHARACTERS
2 min. Withdraw the plate and place it in a current of cold
Appearance
airuntil the excess of chlorine is removed and an area of
White or slightly yellow, crystalline powder.
coatingbelow the points of application gives at most a very
faint blue colour with a drop of potassium iodide and starch Solublllty
solution R. Avoid prolonged exposure to cold air. Spray with Slightly soluble in water, freely soluble in methanol, sparingly
potassium iodide and starch solution R and allow to stand for soluble in ethanol (96 per cent).
5 min. Any spot in the chromatogram obtained with test IDENTIFICATION
solution (a), apart from the principal spot, is not more A. Infrared absorption spectrophotometry (2.2.24).
Comparison cyproheptadine hydrochloride CRS.
reference solution (b) (1.0 per cent). Disregard any spot
remaining at the point of application. B. Dissolve 20 mg in 2 mL of methanol R. The solution gives
reaction (a) of chlorides (2.3.1).
Chlorides (2.4.4)
Dissolve 0.15 g in water R and dilute to 15 mL with the TESTS
same solvent. The freshly prepared solution complies with Acidity
the limit test for chlorides (330 ppm). Dissolve 0.10 g in water R and dilute to 25 mL with the
Phosphates (2.4.11) same solvent. Add 0.1 mL of methyl red solution R. Not more
than 0.15 mL of a 0.40 gIL solution of sodium hydroxide R is
required to change the colour of the indicator.
same solvent. The solution complies with the limit test for
phosphates (100 ppm). Related. substances
Water (2.5.12)
6.0 per cent fa 7.0 per cent, determined on 0.300 g by the Buffersolution Dissolve 6.12 g of patassium dihydrogen
semi-micro determination of water. phosphate R in 900 mL of waterfor chromatography R, adjust
to pH 4.5 with phosphoric acidR and dilute to 1000 mL with
ASSAY
waterfor chromatography R.
Dissolve 0.100 g in 50 mL of a I gIL solution of sodium
hydroxide R in ethylene glycol R and boil under a reflux
condenser for 30 min. Allow to cool and rinse the condenser
mobile phase A.
with 25 mL ofwaterR. Add.75 mL of 2-propanol R, 15 mL
of dilute ni,ricacid R, 10.0 mL of 0.1 M silver nitrate and Reference solution (aJ Dilute 1.0 mL of the test solution to
2.0 mL ofjerrk ammonium sulfare solution RZ and titrate with 100.0 mL with mobile phase A. Dilute 1.0 mL of this
0.1 1\-1 ammonium thiocyanate. solution [0 10.0 mL with mobile phase A.
1 mL of 0./ M st7ver nitrate is equivalent to 13.05 mg of Reference solution (b) Dissolve 2.0 mg of
C,H 15CI,N,O,P. dibenzocycJoheptene CRS (impurity A), 2.0 mg of
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""Em dibenzosuberone CRS (impurity B) and 2.0 mg of
cyprohep'adine impurity C CRS in mobile phase A, add
1.0 mL of the test solution and dilute to 100.0 mL with
mobile phase A.
Cyproheptadine Hydrochloride to 10.0 mL with mobile phase A.
Cyproheptadine Hydrochloride Sesquihydrate Column:
- size: 1::: 0.25 m, 0 = 4.6 mm;
(Cyprohep,adine Hydrochloride IS-Hydrate, Ph. Eur.
monograph 0817) - stationary phase: end-copped oe':ftsi/yl silica gelfor
Mobile phase:
- mo/n7e phase A: acetonitrile for chromatography R, buffer
solution (40:60 VIV);
- mobile phase B: buffer solution, acetonitrile for
chromatography R (40:60 VIV);

(min) (per cent VIV) (per cent VIV)
350.9 41354-29-4
0- 10.0 100 0
Action and use to.O - 10.1 100 --> 0 0-->100
Histamine HI receptor antagonist; antihistamine. 10.1 -.35 0 100
Preparation
Cyproheptadine Tablets Flow rale 1.0 mUmin.
""Em _ Detection Spectrophotometer at 230 nrn.
Injection 10 ~L.
DEFINITION Identification oj impurities Use the chromatogram obtained
4-( 5H-Dibenzo [a,d] [7Jannulen-5-yJidene) -I-methylpiperidine with reference solution (b) to identify the peaks due to
hydrochloride 1.5-hydrate. impurities A, Band C.
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2022 Cyproterone Acetate 1-705
Relativeretention With reference to cyproheptadine

(retention time = about 8 min): impurity C:::; about 0.7;
Cyproterone Acetate
impurity B :;;; about 2.6; impurity A = about 3.9. (Ph. Bur. monograph 1094)
impurity C and cyproheptadine.
Limits:
- impurities A, B, C: for each impurity, not more than
(0.15 per cent); CI
area of the principal peak in the chromatogram obtained 416.9 427-51-0
- total: not more than 5 times the area of the principal peak Action and use
in the chromatogram obtainedwith reference solution (a) Antiandrogen.
(0.5 per cent);
Preparations
- disregard limit: 0.5 times the areaof the principal peak in
Co-cyprindiol Tablets
(0.05 per cent). Cyproterone Tablets
Water (2.5.12) I'I>E<I _

DEFINITION
Sulfated ash (2.4.14) 6-Chloro-3,20-dioxo-1 ~,2~-dihydro-3'H-cyclopropa[1,2]
Maximum 0.1 per cent, determined on 1.0 g. pregna-l,4,6-uien-17-yl acetate.
ASSAY Content
Dissolve 0.250 g in a mixture of 5.0 mL of 0.01 M 97.0 per cent to 103.0 per cent (dried substance).
hydrochlorU: acid and 50 mL of ethanol (96 percem) R. Carry
CHARACTERS
Appearance
inflexion.
I mL of 0.1 M sodium hydroxide is equivalent to 32.39 mg of Solubility
Practically insoluble in water, very soluble in methylene
C2.H"CIN.
chloride, freely soluble in acetone, soluble in methanol,
STORAGE sparingly soluble in anhydrous ethanol.
Protected from light. mp
IMPURITIES About 210 "C.
Specified impun'ties A, B, C. IDENTIFICATION
Firs, identification: A.
Second j'dentificalion: BJ C, D, E.
Comparison cyproterone cuetale CRS.
B. TItin-layer chromatography (2.2.27).
A. 5H-dibenzo[a,d] [7]annuiene (dibenzocycloheptene), examined in methylene chloride R and dilute to 10 mL with
the same solvent.
Reference solution Dissolve 10 mg of cyproterone acetate CRS
in methyle1ie chloride R and dilute to 5 mL with the same
solvent.
Pia,. TLC silica gelFm pia,. R.
MohOe phase cyckJhexane R, ethylaceta,. R (50:50 VW).
Application 5 flL.
B. 10,II-dihydro-5H-dibenzo[a,d] [7]annulen-5-one
(dibenzosuberone), Development Twice over 3/4 of the plate; dry in air between
the 2 developments.
Drying In air.
with the test solution is similarin position and size to the
referencesolution.
C. 5-( l-methylpiperidin-4-yl)-5H-dibenzo[a,d] [7]annuien-5- C. To about I mg add 2 mL of sulfUric add R and heat on a
01. water-bath for 2 min. A red colour develops. Cool. Add this
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'I>E<I
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1-706 Cyproterone Acetate 2022
solution cautiously to 4 mL of water R and shake. - impurities B, C) G: for each impurity, not more than
The solution becomes violet. 0.15 times the area of the principaJ peak in the
D. Incinerate about 30 mg with 0.3 g of anhydrous sodium chromatogram obtained withreference solution (a)
carbonate R over a naked flame for about 10 min. Cool and (0.15 per cent);
dissolve the residue in 5 mL of dilute nitric add R. Filter. - unspecified impurities: for each impurity) not more than
To I mL of the filtrate add I mL of waterR. The solution 0.1 times the area of the principal peak in the
gives reaction (a) of chlorides (2.3.1). chromatogram obtained with reference solution (a)
E. It gives the reaction of acetyl (2.3.1). (0.10 per cent);
TESTS peak in the chromatogram obtained with reference
Specific optical rotation (2.2.7) solution (a) (0.5 per cent);
+ 152 to + 157 (dried substance). - disregard limit: 0.05 times the area of the principal peak in
Dissolve 0.25 g in acetone R and dilute to 25.0 mL with the the chromatogram obtained with reference solution (a)
same solvent. (0.05 per cent).
Liquid chromatography (2.2.29). Maximum 0.5 per cent, determined on 1.000 g by drying at
Test solution Dissolve 10 rng of the substance to be 80 °C at a pressure not exceeding 0.7 kPa.
examined in acetonitrile Rand dilute to 10.0 rnL with the Sulfated ash (2.4.14)
same solvent. Maximum 0.1 per cent) determined on 1.0 g.
Reference solution (aJ Dilute 1.0 mL of me test solutionto ASSAY
100.0 mL with acetonitrile R. Dissolve 50.0 mg in methanol R and dilute to 50.0 mL with
Reference solution (b) Dissolve the contents of a vial of the same solvent. Dilute 1.0 mL of the solution to 100.0 mL
cypro,.rone impurity mixtureCRS (impurities F and I) in with methanol R. Measure the absorbance (2.2.25) at the
1.0 mL of the test solution. absorption maximum at 282 run.
Reference solution (c) Dissolve 2 mg of cyproterone acetate for Calculate the content of C24H29CI04 taking the specific
peak identification CRS (containing impurities B, C, E and G) absorbance to be 414.
in 2.0 mL of acetonitrile R.
STORAGE
Co/umn: Protected from light.
- size: 1= 0.125 m, 0 = 4.6 mm;
- stationary phase: end-<appe4 octadecylsilY/ silica gelfor IMPURITIES
chromatography R (3 urn). Specified impurities B, C, E, F, G.
Mob.e phase ace"'nitrile R, waterR (40:60 VIII). Otherdetectable impurities (thefollowing substances would, if
F/mo rare 1.5 mUmin. present at a sufficient level) be detected by oneor other 0/the tests
in the monograph. They are limited by the general ac.eeptonce
Deuction Spectrophotometer at 254 run. oitenon for otherlunspeclfied impurities andlor by thegeneral
Inje<tion 20 ~L. monograph Substances for phormaceutical US< (2034). It is
Run time Twice the retention time of cyproterone acetate. there/ore not n«essary to identify these impun'tks for
Identification of impurities Use the chromatogram supplied demonsrasion ofcamp/ian«. See also 5.10. Control of impurities
with cyproterone impun'ty mixture CRS and the chromatogram in substances for pharmaceutical use) A) D) JL 1) J.
to impurities F and I; use the chromatogram supplied with
cyproterone aceta,.for peak identification CRS and the
the peaks due to impurities B, C, E and G.
Relativeretention With reference to cyproterone acetate
= =
(retention time about 22 min): impurity E about 0.27;
impurity G = about 0.3; impurity F = about 0.5;
= =
impurity B about 0.7; impurity I about 0.9; A. 3,20-woxo-Ip',2p-dihydro-3'H-cyclopropa [1,2]pregna-
J)4)6-trien-17 -yl acetate)
- resolution: minimum 1.5 between the peaks due [Q
l impurity I and cyproterone acetate.
Limits:
- coreaion facton: for the calculation of content, multiply
the peakareas of the following impurities by the
=
impurity E 0.7;
OCH,
- impuniJI F: not more than 0.4 times the area of the

principal peakin the chromatogram obtained with B. 6-methoxy-3,20-woxo-1 P,2ll-dihydro-3'H-cyclopropa
reference solution (a) (0.4 per cent); [l,2]pregna-IA)6-trien- J7-yJ acetate)
- impurity E: not more than 0.2 times the area of the
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2022 Cysteine Hydrochloride 1-707
o
CI CI
C. 6-cWoro-la-(cWoromethyl)-3,20-dioxopregna-4,6-dien-17- I. 6-cWoro-3,20-dioxopregna-lA,6-trien-17-yl acetate

yl acetate, (delmadinone acetate),
D. lct.-( chloromethyl)-3,6,20-trioxopregn-4-en-17-yl acetate, J. 6a, 7a-epoxy-3,20-dioxo-l p,2P-dihydro-3'H-cyclopropa

[1,2]pregna-I,4-dien-17-yl acetate.
---' PhE"
Cysteine Hydrochloride
o (Cysteine Hydrochloride Monohydrare, Ph. Eur.
monograph 0895)
E. 3,6,20-trioxo-1 P,2Il-dihydro-3'H-cyclopropa [I,2]pregna-
J,4-dien-17-yl acetate,
C,H,CINO,S,H,O 175.6 7048-04-6
H Action and use

Amino acid.
o
PhE" _
CI
DEFINITION
F. 6-cWoro-I7-hydroxy-1 p,2P-dihydro-3'H-cyclopropa (2R)-2-Amino-3-sulfanylpropanoic acid hydrochloride
[l ,2] pregna -1,4,6-triene-3,20-dione, monohydrate.
Product of fermentation or of protein hydrolysis.
Content
CHARACTERS
Appearance
White or almost white, crystalline powderor colourless
crystals.
Solubility
G. 6p-cWoro-7a-hydroxy-3,20-dioxo-1 p,2ll-dihydro-3'H- Freelysoluble in water) slightly soluble in ethanol
cyclopropa[I,2]pregna-I,4-dien-17-yl acetate, (96 per cent).
IDENTIFICATION
First idenufication: A, B, B.
Comparison cysteine hydrochloride monohydrate CRS.
H. 3,20-dioxopregna-l,4-dien-17-yl acetate, C. Thin-layer chromatography (2.2.27).
examined in water Rand dilute to 10 mL with the same
solvent. Add 10 mL of a 40 gIL solution of
N-ethylmaleimide Rin ethanol (96 per emf) R. Allow to stand
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1-708 Cysteine Hydrochloride 2022
for 5 min. Dilute 2 mL of the solution to 10 mL with Reference solution (b) Dissolve 30.0 mg of L-cystine R
water R. (impurity A) in solution A and dilute to 100.0 mL with
Reference solution Dissolve 20 mg of cysteine hydrochloride solution A. Dilute 1.0 mL of the solution to 250.0 mL with
monohydrate CRS in water R and dilute to 10 mL with the solution A.
same solvent. Add 10 ml, of a 40 gIL solution of Reference solution (c) Dissolve 30.0 mg of proline R in
N-ethylmaleimide R in ethanol (96 percenV R. Allow to stand solution A and dilute to 100.0 mL with solution A. Dilute
for 5 min. Dilute 2 mL of the solution to 10 mL with 1.0 mL of the solution to 250.0 mL with solution A.
water R. Reference solution (d) Dilute 6.0 mL of ammonium standard
Plate TLC silica gelplateR. solution (l00 ppm NH<J R to 50.0 mL with solution A. Dilute
kfobl1e phase glacial acetic acidR, waterR, butanol R 1.0 mL of this solution to 100.0 mL with solution A.
(20:20:60 VIVIJI). Reference solution (e) Dissolve 30 mg of isoleucine Rand
Application 5 ~L. 30 mg of leucine R in solution A and dilute to 50 mL with
solution A. Dilute I mL of the solution to 200 mL with
solution A.
Drying At 80 °C for 30 min.
Blank sdution Solution A.
Detection Spray with ninhydrin solution R and heat at 105°C
for 15 min.
Results The principal spot in the chromatogram obtained suitable for the determination of physiological amino acids.
with the test solution is similar. in position, colour and size to
Systemsuitability Reference solution (e):
reference solution.
isoleucine and leucine.
D. Dissolve about 5 mg in 1 mL of dilute sodium hydroxide
solution R. Add I mt. of a 30 gIL solution of sodium
- for impurity A, use the concentration of impurity A in
nitroprusside R. An intense violet colour develops which
becomes brownish-red and then orange. Add 1 mL of
- for any ninhydrin-positive substance detected at 570 om,
hydrochlaric acid R. The solution becomes green.
use the concentration of cysteine hydrochloride
E. Dissolve about 50 mg in 5 mL of water R. Heat to about monohydrate in reference solution (a);
60°C on a water-bath and carefully add, dropwise, 5 mL of - for any ninhydrin-positive substance detected at 440 nm,
strong hydrogen peroxide solution R. Heat the water-bath to use the concentration of proline in reference solution (c);
boiling and maintain the sample on the water-bath for 1 h. if a peak is above the reporting threshold at both
After cooling to room temperature reconstitute the sample to wavelengths, use the result obtained at 570 nm for
10 mL with water R. 2 mL of the solution gives reaction (a) quantification.
of chlorides (2.3.1).
Limits:
TESTS - impun·ty A at 570 nm: maximum 0.5 per cent;
Solution S - any ninhydrin-positive substance: for each impurity,
Dissolve 2.5 g in disu1led waterR and dilute to 50 mL with maximum 0.2 per cent;
the same solvent. - touz1: maximum 1.0 per cent;
Appearance of soludon - reporting threshold: 0.05 per cent.
The solution is clear (2.2.1) and not more intensely coloured The thresholds indicated under Related substances
than reference solution BY6 (2.2.2, Method II). (fable 2034.-1) in the general monograph Substances for
Dilute 10 mL of solution S to 20 mL with water R. pharmaceutical use (2034) do not apply.
Specific optical rotation (2.2.7) Sulfates (2.4.13)
+ 5.5 to + 7.0 (dried substance). Maximum 300 ppm.
Dissolve 2.00 g in hydrochlori< acidR1 and dilute to 25.0 mL Dilute 10 mL of solution S to 15 mL with distilled water R.
with the same acid. Ammonium
Ninhydrin-positive substances Amino acid analysis (2.2.56) as described in the test for
Amino acid analysis (2.2.56). For analysis, use Method 1. ninhydrin-positive substances with the following
Prepare the solutions immediately before use. modifications.
The concentrations of the test solution and the reference Inieaion Test solution, reference solution (d) and blank
solutions may be adapted according to the SQlsitivi£y of the solution.
equipment used. The concentrations of all solutions are Limit:
adjusted so that the system suitability requirements described - ammonium ar 570 nm: not more than the area of the
in general chapter 2.2.46 are fulfilled, keeping the ratios of corresponding peak in the chromatogram obtained with
concentrations between all solutions as described. reference solution (d) (0.02 per cent), taking into account
SouuionA dilute hydrochlorie add Rl or a sample preparation the peak due to ammonium in the chromatogram
buffer suitable for the apparatus used. obtained with the blank. solution.
Test solution Dissolve 30.0 mg of the substance to be Iron (2.4.9)
examined in solution A and dilute to 50.0 mL with Maximum 20 ppm.
solution A. In a separating funnel, dissolve 0.50 g in 10 mL of dilute
Reference solution (a) Dilute 1.0 mL of the test solution to hydrochlori< acidR. Shake with 3 quantities, each of 10 mL,
100.0 mL with solution A. Dilute 2.0 mL of this solution to of methylisobutyl ketone R1, shaking for 3 min each time.
10.0 mL with solution A. To the combined organic layers add 10 mL of water Rand
shake for 3 min. Use the aqueous layer.
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2022 Cystine 1-709
Loss on drying (2.2.32) CHARACTERS

8.0 per cent to 12.0 per cent, determined on 1.000 g by Appearance
drying at a pressure not exceeding 0.7 kPa for 24 h. White or almost white, crystalline powder.
Sulfated ash (2.4.14) Solubillty
Maximum 0.1 per cent, determined on 1.0 g. Practically insoluble in water and in ethanol (96 per cent).
ASSAY It dissolves in dilute solutions of alkali hydroxides.
In a ground-glass stoppered flask dissolve 0.300 g of the IDENTIFICATION
substance to be examined and 4 g of potassium iodide R in First identification: A, B.
20 mL of waterR. Cool the solution in iced water and add Second identification: A, C, D.
3 mL of hydrochloric acidRI and 25.0 mL of O. 05 M iodine.
Stopper the flask and allow to stand in the dark for 20 min.
Titrate with 0.1 Ai sodium thiosulfate using 3 mL of starch B. Infrared absorption spectrophotometry (2.2.24).
solution R, added towards the end of the titration, as Comparison cystine CRS.
indicator. Carry out a blank titration. C. Thin-layer chromatography (2.2.27).
I mL of 0.05 M iodine is equivalentto 15.76 mg of Test solution Dissolve 10 mg of the substance [Q be
C,H.CINO,S. examined in 1 mL of a 103 gIL solution of hydrochloric acidR
STORAGE and dilute to 50 mL with waterR.
Protected from light. Reference solution Dissolve 10 mg of cystine CRS in 1 mL of
a 103 gIL solution of hydrochlcric acidR and dilute to 50 mL
IMPURITIES with water R.
Otherdetectable impun'ties (the following substances would, if
Mobile phase concentrated ammonia R, 2-propanol R
present at a su.fficierll level, be detected by oneor other of the tests
(30:70 VIV).
criterion for other/unspecified impun"ties. It is there/ore not Applicalion 5 ~L.
necessary to idemify these impw;ties for demonstration of Deudopmenc Over 2/3 of the plate.
compliance. See also 5.10. Control of impurities in substances for Drying In air.
pharmaceutical use) B. Detection Spray with ninhydrin solution R and heat at 105°C
for 15 min.
H NH2
Hn~c' sV Results The principal spot in the chromatogram obtained
-.rl ~s/ ............... C~H with the test solution is similar in position, colour and size [0
H Nt<,
the principal spot: in the chromatogram obtained with the
reference solution.
A. (2R,2'R)-3,3'-disulfaoediylbis(2-aminopropaooic acid)
D. To 0.1 g carefully add I mL of slTOIIg hydrogen peroxide
(cystine),
solution Rand 0.1 mL of/em. chloride sduuon RI. Allow to
H NH2 cool. Add I mL of dilute hydrochloric acid Rand 5 mL of
HO V waterR. Add I mL of barium chloride solution RI. Turbidity
~Co,H or a white precipitate develops within 3 min.
B. (2S)-2-amino-3-hydroxypropaooic acid (serine). TESTS

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhEll Appearance of solution
than reference solution Y, (2.2.2, Metlwd II).
Dissolve 1.0 g in dilute hydrochloric acidR and dilute to
10 mL with the same acid.
Cystine Specific optical rotation (2.2.7)
(Ph. Bur. monograph 0998) -224 to -218 (dried substance).
Dissolve 0.50 g in a 103 gIL solution of hydrochloric acid R
Amino acid analysis (2.2.56). For analysis, use Method 1.
The concentrations of the test solution and the reference
CJf12N,O,S, 240.3 56-89-3 solutions may be adapted according [0 the sensitivity of the
equipment used. The concentrations of all solutions are
Action and use adjusted so that the system suitability requirements described
Amino acid. in general chapter 2.2.46 are fulfilled, keeping the ratios of
PhEIl _ concentrations between all solutions as described.
Solulion A A 10.3 gIL solution of hydrochloric acid Ror a
DEFINITION sample preparation buffer suitable for the apparatus used.
L-Cystine (3,3'-disulfaoediylbis[(2R)-2-aminopropaooic
acid]).
Product of fermentation or of protein hydrolysis. solution A.
Content
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1-710 Cystine 2022
Reference solution (a) Dilute 1.0 mL of the test solution to me peak due to anunonium in the chromatogram
100.0 mL with solution A. Dilute 2.0 mL of thissolutionto obtainedwith the blank solution.
10.0 mL with solution A. Iron (2.4.9)
Reference solution (b) Dissolve 30.0 mg of proline R in Maximum 10 ppm.
solution A and dilute to 100.0 mL with solution A. Dilute In a separating funnel, dissolve 1.0 g in 10 mL of dl1ute
1.0 mL of the solution to 250.0 mL with solution A. hydrochloric add R. Shake with 3 quantities, each of 10 mL,
Reference solution (c) Dilute 6.0 mL of ammonium standard of methylisobutyl ketone RJ, shaking for 3 min each time.
solution (l00 ppm NH.J R to 50.0 mL with solution A. Dilute To the combined organic layers add 10 mL of water Rand
1.0 mL of this solution to 100.0 mL with solution A. shake for 3 min. Use the aqueous layer.
Reference solution (d) Dissolve 30 mg of isoleucine Rand Loss on drying (2.2.32)
30 mg of leucine R in solution A and dilute to 50.0 mL with Maximwn 0.5 per cent) determined on 1.000 g by drying in
solution A. Dilute 1.0 mL of the solution to 200.0 mL with an oven at 105 "C.
solution A. Sulfated ash (2.4.14)
Blank solution Solution A. Maximum 0.1 per cent, determined on 1.0 g.
ASSAY
solutions (a), (b) and (d) into the amino acid analyser. Run a
In a flask with a ground-glass stopper, dissolve 0.100 g in a
program suitable for the determination of physiological
mixture of 2 mL of dilute sodium hydroxide solution Rand
amino acids.
10 mL of water R. Add 10 mL of a 200 gIL solution of
System suitability Reference solution (d): potassium bromide R, 50.0 mL of O. 0167 M potassium bromate
- resolution: minimum 1.5 between the peaks due to and 15 mL of dihue hydrochloric acidR. Stopper the flask and
isoleucineand leucine. cool in iced water. Allow to stand in Ute dark for 10 min.
Calculau"on of percentage contents: Add 1.5 g of potassium iodide R. After 1 mini titrate with
- for anyninhydrin-positive substance detected at 570 nm, 0.1 M sodium thiosulfate, using 2 mL of starch solution R,
use the concentration of cystine in reference solution (a); added towards the end-point, as indicator. Carry out a blank
- for any ninhydrin-positive substance detectedat 440 run, titration.
use the concentration of proline in reference solution (b); 1 mL of 0.0167 M potassium bromate is equivalent to
if a peak is above the reporting threshold at both 2.403 mg ofC.H. 2N 20.S2 •
wavelengths, use the result obtained at 570 nm for
quantification. STORAGE
Limits:
- any ninhydn'n-positive substance: for each impurity, IMPURITIES
maximum0.2 per cent; Otherdetutable impurities (thefollowing substances would, if
- ICtal: maximum 0.5 per cent; present at a sufficient level) be detected by one or other of the tests
- reponing threshold: 0.05 per cent. in the monograph. They are limited by the general acceptance
The thresholds indicated underRelated substances crililnan for atherlunspedfied impurities. It is therefore nat
(Table 2034.-1) in the general monograph Substances for n«essary W identify these impuniies for demonstration 0/
pharmaceutical use (2034) do not apply. compliance. See also5.10. Confl'Ol of impurities in substances for
pharmaceutical use) A.
Chlorides
Maximum 200 ppm.
Dissolve 0.5 g in 5 mL of dilute nitn·c acid R and dilute to
10 mL with the same solvent. Add 10 mL of strong hydrogen
peroxide solution R and heat on a water-bath for 30 min. Cool
and dilute to 50 mL with water R. Add 1 mL of silver nitrate
A. L-tyrosine «2S)-2-amino-3-(4-hydroxyphenyl)propanoic
solution R2 and mix. Allow to stand for 5 min protected from
light. Any opalescence in the solution is not more intense acid).
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
than that in a standard prepared at the same time and in the
same manner using 2 mL of chloride standard solution (50 ppm
CD R. Examine the tubes laterally against a black
background.
Sulfates (2.4.13)
Maximum 300 ppm.
Dissolve 0.5 gin 5 mL of dilutehydrochloric acidR and dilute
to 15 mL with distilkd water R.
Ammonium
Amino acid analysis (2.2.56) as described in the test for
modifications.
Injuuf:m Test solution, reference solution (c) and blank
solution.
Limit.
- ammonium at 570 nm: not more than the area of the
reference solution (c) (0.02 per cent), taking into account
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2022 Cytarabine 1-711
Cytarabine *** Reference solution (b) Dissolve 2 mg of uridine R

*** *** (impurity B) and 2.5 mg of me substance to be examined in
(ph. Bur. monograph 0760) *** water R and dilute to 100 mL with the same solvent.
Column:
- size: 1= 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped IXtadecylsi1YI silica gelfor
Mobile phase:
- mobile phase A: methanol R, buffer solution (2:98 VIr);
- mobile phase B: methanol R, buffer solution (30:70 VIr);

(mJn) (per cent VIJI) (per cent VIJI)
243.2 147-94-4
0-10 100 0
Action and use 10 - 20 100 -+ 0 0-+100
Pyrimidine analogue; cytotoxic. 20·25 0 100
Preparation
Cytarabine Injection FWw rate 1.0 mk/mln.
PhE" _
Injection 20 ~L.
DEFINITION Identification of impun"ties Use the chromatogram obtained
4-Amino-I-~D-arabinofuranosylpyrimidin-2(1H)-one. with reference solution (a) to identify the peak due to
Content impurity A; use the chromatogram obtained with reference
99.0 per cent to 101.0 per cent (dried substance). solution (b) to identify the peak due to impurity B.
CHARACTERS Relatwe retention With reference (Q cytarabine (retention
Appearance time = about 9 min): impurity B ;:;;: about 1.2;
impurity A ;:;;: about 1.7.
Solubility
Freelysoluble in water, practically insoluble in ethanol
cytarabine and impurity B.
Calculation of percentage "",tents:
mp
- for each impurity, use the concentration of impurity A in
About 215 "C. reference solution (a).
Infrared absorption spectrophotometry (2.2.24). - impun'ty A: maximum 0.15 per cent;
Comparison cytarabine CRS. - unspecified impurities: for each impurity, maximum
0.05 per cent;
TESTS
than reference solution v, (2.2.2, Method If). Loss on drying (2.2.32)
Dissolve 1.0 g in water Rand dllute to 10 mL with the same
V"'UQ at 60 °C at a pressure of 0.2-0.7 kPa for 3 h.
solvent.
Dissolve 0.250 g in water R and dilute to 25.0 mL with the ASSAY
same solvent. Dissolve 0.200 g in 60 mL of anhydrous acetic acidR,
warming if necessary. Titratewith 0.1 M perchloric acid,
Related substances
determining the end-point potentiometrically (2.2.20).
I mL of 0.1 M perchlori<: acid is equivalent to 24.32 mg of
Buffer so/mum Dissolve 1.38 g of sodium dihydrogen phosphate
monohydrate Rand 1.42 g of anhydrous disodium hydrogen C9H13N305'
phosphate R in 950 mL of waterfor chroma",graphy R, adjust STORAGE
to pH 7 with a 4 gIL solution of sodium hydroxide Rand In an airtight container, protected from light.
dilute to 1000 mL with waterfor chroma"'graphy R. IMPURITIES
Testsolution Dissolve 25.0 mg of the substance to be Specified imputities A.
examined in waler R and dilute to 10.0 mL with the same
Otherdetectable impun'/ies (thefollowing substances would, if
solvent.
present at a sufficientlwel, be detected by oneor other of the tests
Reference solution (a) Dissolve 5.0 mg of uracil in the monograph. They are limited by thegeneral acceptance
arabinoside CRS (impurity A) in water R and dilute to cruenon for otherhmspecified impuriti<s and/or by thegeneral
100.0 mL with the same solvent. Dilute 0.5 mL of the monograph Substances for pharmaceutical use (2034). 11 is
solution to 10.0 mL with water R. therefore not necessary to identify these impurities for
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1-712 Dacarbazine 2022
demonstration of compliance. See also 5.10. Control of impurities NH,

AvcH'
0'0
in substances for pharmaceutical use) B} CJ D, E, F, G, H, J.
N.)
o
HNJ
HOV~~ OH OH
POH G.4-amino-5-methyl-I-I3-D-ribofuranosylpyrimidin-2(1H)-
one (5-methylcytidine),
A. 1-~-D-arabinofuranosylpyrimidine-2,4(IH,3H)-dione
NH
(uracil arabinoside),
))
HHt-t H
HO~O
H ''-OH
H. (2R,3R,3aS,9aR)-2-(hydroxymethyl)-6-imino-2,3,3a,9a-
tetrabydro-6H-fum [2',3 ':4,5) [1,3] oxazolo[3,2-0)
pyrimidin-3-o1,
B. 1-l3-o-ribofuranosylpyrimidine-2,4(IH,3H)-dione NH,
(uridine),
~CH,
N.)
HOVQ
POH
C. 4-aminopyrimidin-2(1H)-one (cytosine),
1. 4-amino-I-l3-o-arabinofuranosyl-5-methylpyrimidin-2
(lH)-one.
~ ~ Ph&
D. pyrimidin-2,4(IH,3H)-dione (uracil), Dacarbazine

~~ OH OH
182.2 4342-03-4
E. 4-amino-I-~-D-ribofuranosylpyrimidin-2(1H)-one
(cytidine), ,,
Action and use
Cytotoxic alkylating agent.
PhEw _
DEFINITION
5-[(1 B)-3,3-D imethyltriaz-I-en-I-yl]-I H-imidazole-4-
carboxamide.
Content
CHARACTERS
F. 4-amino-I-(2,5-anhydro-~-o-arabinofuranosyl)pyrimidin-2 Appearance
(IH)-one, White or slightly yellowish, crystalline powder.
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2022 Dacarbazine 1-713
Solubility Reference solution (d) Dissolve 5.0 mg of dacarbazine

Slightly soluble in water and in anhydrous ethanol, practically impurity B GRS in distilled wow R and dilute to 50.0 mL
insoluble in methylene chloride. with the same solvent.
IDENllFICATION Reference solution (e) Dilute 1.0 mL of reference solution (d)
First identification: B. to 10.0 mL with distilled waterR.
Second identification: A, C. Reference solution (f) Mix I mL of reference solution (a) and
I mL of reference solution (d) and dilute to 10 mL with
distilled wow R.
(2.2.25).
Column:
Test solution Dissolve 15.0 mg in a 10.3 gIL solution of
hydrochloric acidR and dilute to 100.0 mL with the same
=
- size: 1= 0.25 m, 0 4.6 mm;
- stationary phase: end-capped ocradecylsilyl silica gelfor
solution. Dilute 5.0 mL of the solution to 100.0 mL with a
10.3 gIL solution of hydrochloric acid R.
Mobile phase 15.63 gIL solution of glacial acetic acidR
Spectral range 200-400 mo.
containing 2.33 gIL of sodium diocty/ sdfosuccinate R.
Absorption maximum 323 om.
Shoulder 275 nm.
Injection 25 ul, of the test solution and reference
B. Infrared absorption spectrophotometry (2.2.24). solution (c).
Comparison dacarbazine CRS. Rim time 3 times the retention time of impurity A.
C. Thin-layer chromatography (2.2.27). identification of impurities Use the chromatogram obtained
Test solution Dissolve 2.0 mg of the substance to be with reference solution (c) to identify the peak due to
examined in methanol R and dilute to 5.0 mL with the same impurity A.
solvent. Retention time Impurity A = about 3 min.
Reference solution Dissolve 2.0 mg of dacarbazine CRS in Limits:
methanol R and dilute to 5.0 mL with the same solvent. - impurity A: not more than the area of the
Plow TLC silica gelF254 plow R. corresponding peak in the chromatogram obtained
Mobile phase glacial acetic acid R, walerR, butanol R with reference solution (c) (0.2 per cent);
(10:20:50 VIVIV). - unspecified impurities eluting afterimpun'cy A: for each
Application 10 ~L. impurity, not more than 0.5 times the area of the
Drying In air.
B. Liquid chromatography (2.2.29) as described in test A for
Detection Examine in ultraviolet light at 254 run. related substances with the following modifications.
Results The principal spot in the chromatogram obtained Mobile phase Mix 45 volwnes ofa 15.63 gIL solution of
with the test solution is similar in position and size to the glacial acetic acid R containing 2.33 gIL of sodium dioctyJ
principal spot in the chromatogram obtained with the sulfosuccinate Rand 55 volumes of methanol R.
reference solution.
Injection 10).lL of the test solution and reference
TESTS solutions (b), (e) and (I).
Appearance of solution Run time Twice the retention time of dacarbazine.
Identification 0/ impurities Use the chromatogram obtained
than reference solution BY6 (2.2.2, Method II).
with reference solution (e) to identify the peak due to
Dissolve 0.25 g in a 210 gIL solution of dtn', add impurity B.
monohydrate R and dilute to 25.0 mL with the same solution. Relative retention With reference to dacarbazine (retention
Related substances time = about 27 min): citric acid = about 0.05;
A. Liquid chromatography (2.2.29). Use freshly prepared impurity B = about 0.8.
sdutions and protect themfrom light. Use freshly prepared mobile System suiuwl1ity Reference solution (0:
phaseas it contains sodium diocty/ sulfosucdnate. - resolution: minimum 1.5 between the peaks due to
Flush the column with a mixture of equal volumes of impurity Band dacarbazine.
methanol R and waterfor chromatography R for at least 2 h at Limits:
the end of each day or after all tests have been completed. - impurity B: not more than the area of the
Test solution Dissolve 50.0 mg of the substance to be corresponding peak in the chromatogram obtained
examined and 75 mg of citric add monohydrate R in distilled with reference solution (e) (0.1 per cent);
waterR and dilute to 5.0 mL with the same solvent. - unspecified impurities: for each impurity, not more than
Reference sdution (a) Dilute 1.0 mL of the test solution to the area of the peak due to dacarbazine in the
100.0 mL with distilled warer R. chromatogram obtained with reference solution (b)
Reference solution (b) Dilute 1.0 mL of reference solution (a) (0.10 per cent);
to 10.0 mL with distilled waterR. - total: not more than 5 times the area of the peak due
to dacarbazine in the chromatogram obtained with
Reference sduuon (c) Dissolve 5.0 mg of dcuarbazine
impurity A GRS in distilled wow R and dilute to 50.0 mL
- disregard limt"t: 0.5 times the area of me peak due to
with the same solvent. Dilute 5.0 mL of this solution to
dacarbazine in the chromatogram obtained with
25.0 mL with distilled wow R.
reference solution (b) (0.05 per cent); disregard the
peak due to citric acid.
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1-714 Dalteparin Sodium 2022
Impurity D demonstration of compliance. See also 5.10. Control of impurities

Gas chromatography (2.2.28). Prepare the solutions immediately in substances for pharmaceutical use) C.
before use.
Test solution Suspend 0.500 g of the substance to he
examined in 5.0 mL of ethyl acetate Rl. Filter through a
membrane filter (nominal pore size 0.45 ....m).
Reference solution Dilute 1.250 g of a 40 per cent mlm
solution of dimethylamine R (equivalent to 0.500 g of
A. 3,7-dihydro-4H-imidazo[4,5-d] [1,2,3]triazin-4-one
impurity D) to 100.0 mL with ethyl ace""e RI. Dilute
1.0 mL of the solution to 100.0 mL with ethyl acetate RI. (2-azahypoxanthine),
Column:
- size: 1= 30.0 m, 0 = 0.53 mm;
- stationary phase: base-deactivated macrogol R (film thickness
1.0 urn).
Cartiergas helium for chromatography R. B. 5-amino-1H-imidazole-4-carboxamide,
Flow rale 3.0 mUmin. ;=N
Split ratio 1:10. HNyYN~
Temperature:
HN",N 0
Time Temperature
(min) CC) C. 5-diazenyl-1H-imidazole-4-carboxamide,
Column O· I 40
I ·6 40 50
6 - 12 50 200
12 - 22 200
Injection port 180
D. N-methylmethanamine (dimethylamine).
~ ''llE1l
Detector 220

Injection 1.0 ~L.
Retention time Impurity D = about 3 min.
Dalteparin Sodium
System suitability Reference solution: (Ph. Bur. monograph J/95)
'O~
5.0 per cent for the areaof the peak due to impurity D
determined on 6 injections.
NaOsS °
R2 HO_
Calculation of percentage content.
- use the concentration of impurity D in the reference o OH
o
solution.
Limit:
- impun"ty D: maximum 0.05 per cent.
Water (2.5.12)
Maximum 0.5 per cent, determined on 1.00 g. O .... sosNa
Sulfated ash (2.4.14) n"'3Io20. R"'HorSOsNa. R''''SOsNaorCo-CHs
Maximum 0"1 per cent, determined on 1.0 g. R2 .: H and R3 '" C0 2Na Of R2 =C<>,:Na andR3 '" H
ASSAY
Action and use
with 0.1 M perchloric acid, determining the end-point Low molecular weightheparin.
potentiometrically (2.2. 2tJ). Preparation
1 mL of 0.1 M perchlotic acid is equivalent to 18.22 mg Dalteparin Sodium Injection
of Co;H,oN.O. PhEIl _
STORAGE DEFINITION
Protected from light, at a temperature of 2 °C to 8°C. Dalteparin sodium is the sodium salt of a low-molecular-
IMPURITIES mass heparin that is obtained by nitrous acid
Specified impurities A, B, D. depolymerisation of heparin from porcine intestinal mucosa.
Otherdetectable impurities (the folWwing sttbstances could, if The majority of the components have a 2-o-sulfo-«-L-
present at a sufficient level, be detected by oneor other of the tests idopyranosuronic acid structure at the non-reducing end and
in the monograph. They are limitedby thegeneral acceptance a 6-0-sulfo-2,5-anhydro-o-mannitol structure at the reducing
criterion for othemmspedfiedimpurities andlor by the general end of theirchain.
monograph Substances for pharmaceutical use (2034). It is Dalteparin sodium complies with the monograph Low-mol«Uiar-
therefore not necessary to identify these imptmues for mass heparins (0828) with the modifications and additional
requirements below.
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2022 Dalteparin Sodium 1-715
The mass-average relative molecular mass ranges between - as mobile phase at a flow rate of 1.0 mUmin a solution
5600 and 6400, with a characteristic value of about 6000. consisting of 13.61 g of sodium acetare R dissolved in
The degree of sulfataticn is 2.0 to 2.5 per disaccharide unit. water R, adjusted to pH 4.3 with phosphoric add Rand
diluted to 1000 mL with water R;
The potency is not less than 110 ill and not more than
- 3S detector an appropriate electrochemical device with the
210 ill of anti-factor Xa activity per milligram, calculated
following characteristics and settings: a suitable working
with reference to the dried substance. The anti-factorTIa
electrode, a detector potential of + 1.00 V versus AglAgCl
activity is not less than 35 IU/mg and not more than
reference electrode and a detector sensitivity of 0.1 J1A full
100 ill/mg, calculated with reference to the dried substance.
scale.
The ratio of anti-factor Xa activity to anti-factor IIa activity is
between 1.9 and 3.2. inject 100 ~L of reference solution (d). When the
chromatograms are recorded in the prescribed conditions, the
PRODUCTION retention time for nitrite is 3.3 to 4.0 min. The test is not
Dalteparin sodium is produced by a validated manufacturing valid unless:
and purification procedure under conditions designed to - the number of theoretical plates calculated for the nitrite
minimise the presence ofN-NO groups. peak is at least 7000 per metre per column (dalteparin
The manufacturing procedure must have been shown to sodium will block the binding sites of the stationary
reduce any contamination by N-NO groups to approved phase, which will cause shorter retention times and lower
limits using an appropriate, validated quantification method. separation efficiency for the analyte; the initial
IDENTIFICATION performance of the column may be partially restored
using a 58 gIL solution of sodium chloride R at a flow rate
Carry out identification test A as described in the monograph
of 1.0 mlJmin for I h; after regeneration the column is
Low-mo/ecular-mass heparins (0828) using da/teparin
rinsed with 200 mL to 400 mL of waterR);
sodium CRS.
- the symmetry factor for the nitrite peak is less than 3;
Carry out identification test C as described in the monograph - the relative standard deviation of the peak area for nitrite
Low-mokcular-mass heporins (0828). The following obtained from 6 injections is less than 3.0 per cent.
requirements apply.
inject 100 ~L each of reference solutions (c) and (e).
The mass-average relative molecular mass ranges
The test is not valid unless:
between 5600 and 6400. The mass percentage of chains - the correlation factor for a linear relationship between
lower than 3000 is not more than 13.0 per cent. The mass concentration and response for reference solutions (c), (d)
percentage of chains higher than 8000 ranges between and (e) is at least 0.995;
15.0 per cent and 25.0 per cent. - the signal-to-noise ratio for reference solution (c) is not
TESTS less than 5 (if the noise level is too high, electrode
Appearance of solution recalibration is recommended);
Dissolve 1 gin 10 mL of waterR. The solution is clear - a blank injection of waterR does not give rise to spurious
(2.2.1) and not more intensely coloured than intensity 5 of peaks.
the range of reference solutions of the most appropriate Inject 100 J,lL of the test solution. Calculate the content of
colour (2.2.2, Method If). nitrite from the peak areas in the chromatogram obtained
Nitrite with reference solutions (c), (d) and (e).
Not more than 5 ppm. Examine by liquid chromatography Boron
(2.2.29). Rinse aU 'lIOiumetricf/asks at least three times with Not more than 1 ppm, determined by inductively coupled
waterR before the preparation of the solutions. plasma atomic emission spectroscopy.
Testsolution Dissolve 80.0 mg of the substance to be Boron is determined by measurement of the emission from
examined in water R and dilute to 10.0 mL with the same an inductively coupled plasma (ICP) at a wavelength specific
solvent. Allow to stand for at least 30 min. to boron. The emission line at 249.733 nm is used. Use an
Reference solution (aJ Dissolve 60.0 mg of sodium nitrite R in appropriate apparatus, whose settings have been optimised as
water R and dilute to 1000.0 mL with the same solvent. directed by the manufacturer.
Forthe preparation of reference solutiDn (b), use a piPe/ie Test solution Dissolve 0.2500 g of the substance to be
prevUiusly rinsed with reference solution (a). examined in about 2 mL of waterfor chromatography R, add
Reference solution (b) Dilute 1.00 mL of reference 100 ~L of nitric acid R and dilute to 10.00 mL with the same
solution (a) to 50.0 mL with walerR. solvent.
Before preparing reference solutions (c), (d) and (e), rinse all Reference solution (aJ Prepare a 1 per cent VIV solution of
pipeltes with reference solution (b). nitric acidR in waterfor chromawgraphy R (blank).
Reference solutiDn (c) Dilute 1.00 mL of reference Reference solutio" (b) Prepare a 11.4 ~g/mL solution of boric
solution (b) to 100.0 mL with waterR (corresponding to add R in a I per cent VIV solution of nitric acid R in water for
I ppm of nitrite in the test sample). chromawgraphy R (STD,.,).
Reference solution (d) Dilute 3.00 mL of reference Reference solution (c) Dissolve 0.2500 g of a reference
solution (b) co 100.0 mL with Waler R (corresponding to dalteparin sodium with no detectable boron in about 2 mL of
3 ppm of nitrite in the test sample). waterfor chromatIJgraphy R, add 100 ~L of nitric acid Rand
dilute to 10.00 mL with the same solvent (STOo).
Reference sdution (e) Dilute 5.00 mL of reference
solution (b) to 100.0 mL with waterR (corresponding to Reference solution (d) Dissolve 0.2500 g of a reference
5 ppm of nitrite in the test sample). dalteparin sodium with no boron detected in about 2 mL of
a 1 per cent V/V solution of nitric acidR in waterfor
The chromatographic procedure may be carried out using:
chromatography R, add 10 ~L of a 5.7 mg/mL solution of
- a column 0.125 m long and 4.3 mm in internal diameter
packed with a strong anion-exchange resin;
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1-716 Danaparoid Sodium 2022
boric acidR and dilute to 10.00 mL with the same soJvent IDENTIFICATION
(STD I ) . This solution contains 1 ~g/mL of boron. A. The ratio of anti-factor Xa activity [0 anti-factor lIa
Calculate the content of boron in the substance to be activity, determined as described under Assay and Tests
examined, using the following correction factor: respectively, is not less than 22.
B. Molecular mass distribution (see Tests): the mass-average
f= (STD I-STDo)x2 relative molecular mass ranges between 4000 and 7000.
(STD o" - blank) TESTS
pH (2.2.3)
5.5 to 7.0.
drying in an oven at 60°C over diphosphorus pemoxide R at a Dissolve 0.5 g of the dried substance to be examined in
pressure not exceeding 670 Pa for 3 h. carbon dioxide-free waterR and dilute to 50 mL with the same
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Pl>E<r solvent.
Antl-faceor IIa activity
Maximum 0.5 units per milligram (dried substance).
Cany out the testat room temperature. Human thrombin
Danaparoid Sodium sdution Rl must be kept on ice until use.
Test solutwtlS Prepare 2 independent series of dilutions in
(Ph. Eur. monograph 2090) geometric progression of the substance to be examined in
phosphate buffer solution pH 6.5 R and in.the concentration
Chondroitin sutrala and Heparan suflale famlfy range of 0.0005-0.005 units of anti-factor Ila activity per
dermatan sultate family OR. millilitre.
~H
OR1
Reference solutions Prepare 2 independent series of dilutions
c~~'fl H
CD,Na
00
0
0
H
in geometric progression of danaparoid sodium CRS in
phosphate buffer solution pH 6.5 R and in the concentration
range of 0.0005-0.005 units of anti-factor Ila activity per
OH OH RS.... NH
0yNH HO rnillilitre.
OR3 CH3 OR6
Transfer 50 ~ of each solution into the wells of a 96-well
microtitre plate. To each well add 50 ~L of antithrombin III
R1, R2, R3, R4, R6 =H or SO~a solution R3 and 50 ~ of human thrombin solution RI. Shake
R5 =CG-CH; or S03Na the microtitre plate but do not allow bubbles to fonn.
Incubate for 75 min. To each well add 50 JlL of chromogenic
substrate R4. Shake the microtitre plate. Measure the
Action and use absorbance at 405 run (2.2.25) using a suitable reading
Heparinoid; prevention of deep vein thrombosis. device, exactly 2 min after the addition of the chromogenic
substrate. Measure the absorbance again a' 405 run (2.2.25),
Pl>E<r _
exactly 22 min after the addition of the chromogenic
DEFINITION substrate. Use M (A22 - A 2 ) for the calculation of the anti-
Preparation containing the sodium salts of a mixture of factor IIa activity. Determine the blank amidolytic activity in
sulfated g1ycosaminoglycans present in porcine tissues. a similar manner, using phosphate buffersolution pH 6.5 R as
Danaparoid sodium is prepared from the intestinalmucosa of the blank solution (minimum 8 blanks per microtitre plate).
pigs. Its major constituents are heparan sulfate and dermatan Calculate the activity of the substance to be examined
sulfate. On complete hydrolysis it liberates n-glucosamlne, in units of anti-factor Ila activity per milligram using a
n-galaetosamine, n-glucuronic acid, L-iduronic acid, acetic suitable statistical method, for example the parallel-line assay
(5.3).
acid and sulfuric acid. It has the characteristic property of
enhancing the inactivation of activated factor X (factor Xa) Chondroitin sulfate and dermatan sulfate
by antithrombin. It has a negligible effect on the inactivation Chondroitin sulfate: maximum 8.5 per cent (dried
rate of thrombin by antithrombin. substance); dermatan sulfate: 8.0 per cent to 16.0 per cent
Potency (dried substance).
11.0 to 19.0 anti-factor Xa units per milligram (dried Determine by selective enzymatic degradation.
substance). Test solutions Dry the substance to be examined at 60 °C
PRODUCTION over diphosphorus pentoxide R at a pressure of about 670 Pa
for 3 h. Dissolve 0.200 g of the dried substance in 10.0 mL
The animals from which danaparoid sodium is derived must
of water R. Dilute this solution as necessary to obtain 3 test
fulfil the requirements for the health of animals suitable for
solutions containing 20 mg/mL, 10 mg/mL and 5 mglmL of
human consumption. Danaparoid sodium is prepared using a
the dried, substance to be examined in waterR.
. process that ensures that the relative proportion of active
sulfated giycosaminoglycans is consistent. It is produced by Chondroitin sulfate reference solutions Dry chondroitin sulfate
methods of manufacnuing designed [0 minimise or eliminate sodium CRS over diphospho7US pemoxide R at room
endotoxins and hypotensive substances. temperature at a pressure of about 670 Pa for 16 h. Prep-are
solutions containing 1 mg/mL, 2 mg/mL and 3 mg/mL of
CHARACTERS dried chondroitin sulfate sodium CRS in waterR.
Appearance
Dermaton sulfate reference solutions Dry dermaran sulfate CRS
White or almost white, hygroscopic powder.
over diphosphorus pemoxide R at room temperature at a
Solubility pressure of about 670 Pa for 16 h. Prepare solutions
Freely soluble in water.
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2022 Danaparoid Sodium 1-717
containing I mglmL, 2 mglmL and 3 mglmL of dried I) y-intercept of the curve obtained with the chondroitin sulfate
reference solutions with chondroitinase ACj
dermtuan sulfate CRS in water R. /2 y-Intercepr of the curve obtained with the cbondroitin sulfate
Chondroitinase ABC solution Dissolve chondroitinase ABC R reference solutions with cbondroitinase ABC;
in ens-sodium acetate-sodium chloride buffer solution pH 8.0 R to /) y-intercept of the curve obtained with the dermatan sulfate
reference solutions with chendrohinase ABC.
obtain an activity of 0.5-1.0 units per millilitre.
Chondroitinase AC solution Dissolve chondroitinase AC R in Calculate the average percentage content of chondroitin
iris-sodium aceuue-sodiem chloride bufftrsolution pH 7.4 R to sulfate in the test solutions for aU tested concentrations using
obtain an activity of 1.0-2.0 units per millilitre. the following expression:
Procedure:
- Degradation with chondroitinase ABC. Label 2 sets of
10 tubes in triplicate: TI, T2 and T3 for the test
solutions; SD 1, SD2 and SD3 for the derrnatan sulfate
reference solutions; SC 1, SC2 and SC3 for the Molecular mass distribution
chondroitin sulfate reference solutions; and B for the Size-exclusion chromatography (2.2.30).
blank (waler R). To each tube add 1.25 mL of Iris-sodium Test solution Dissolve 10 mg of the substance to be
acetale buffer solution pH 8.0 Rand 150. ~L of the lest examined in 2 mL of the mobile phase.
solutions, dermatan sulfate reference solutions,
Reference solution Dissolve 10 mg of danaparoid sodium GRS
chondroitin sulfate reference solutions or water R.
in 2 mL of the mobile phase.
To each tube in I set of tubes add 75 ~L of
chondroitinase ABC solution. To determine the blank Column:
response level, add 75 !!L of tns-sodium acetate-sodium =
- size: 1= 0.60 m, 0 7.5 mm;
chloride buffer solution pH 8.0 R to each tube in the other - stationary phase: hydrophilic silica gelfor chromatography R
set of tubes. Mix the contents of the tubes using a vortex (10 IlID) with a fractionation range for proteins with a
mixer, cover with appropriate stoppers and incubate at relative molecular mass of approximately 5000 to
37°C for at least 24 h. 100000;
- Degradation with chondroitinase AG. Label 7 tubes in - temperature: 30°C.
triplicate: TI, T2 and T3 for the test solutions; SCI, SC2 Mobil, phase 28.4 gIL solution of anhydrous sodium sulfate R
and SC3 for the chondroitin sulfate reference solutions; adjusted 10 pH 5.0 with dilule sulfuric acidR.
and B for the blank (water 1<). To each tube add 1.25 mL Flow rate 0.9 mUmin ± 2 per cent.
of tris-sodium acetate buffer solution pH 7.4 R and ISO ilL Detection Spectrophotometer at 210 om.
of the test solutions, '
Injmion I00 ~L.
chondroitin sulfate reference solutions or water R.
Add 75 ~ of chondroitinase AC solution to each tube. Run time For a period of time ensuring complete elution of
Mix the contents of the tubes using a vortex mixer, cover sample and solvent peaks (about 40 min).
with appropriate stoppers and incubate at 37°C for at Systemsuitability Inject me reference solution twice; the
least 24 h. difference between the retention times corresponding to the
After the incubation period mix the contents of the tubes maxima of the peaks is not more than 5 s.
using a vortex mixer and dilure to 12 times with water R. Calihratitm Calibration is achieved by taking the relevant
Measure the absorbances (2.2.25) of the diluted solutions at part of the chromatogram obtained with the reference
234 om against water R using a suitable spectrophotometer. solution, i.e. excluding the sharp peak at the end of the
Calculaticm Calculate the mean blank absorbance of each chromatogram, and matching the chromatogram obtained
reference solution, i.e. the mean of the ebsorbances of the with the test solution with the calibration table obtained with
reference solutions to which no chondroitinase ABC has been the reference solution. From the calibration curve obtained,
added. Subtract the mean blank absorbance value from the determine the molecular mass distribution of the sample.
individual absorbance of each reference solution. Calculate A calibration table is supplied with danaparoid sodium GRS.
linear regression curves for the chondroitin sulfate reference Limits:
solutions and the dermatan sulfate reference solutions by - chains with a relative molecular mass less than 2000:
plotting the blank-corrected absorbances against the maximum 13 per cent;
concentrations. - chains with a relative molecular mass less than 4000:
Calculate the average percentage content of dermatan sulfate maximum 39 per cent;
in the test solutions for all tested concentrations using me - chains with a relative molecular mass between 4000 and 8000:
following expression: minimum SO per cent;
- chains with a relative., molecular mass higher than 8000:
CA, -AI -I.) xB, maximum 19 per cent;
A, -AI - chains with a relative molecular mass higher than 10000:
BI.
maximum 11 per cent.
B, xG
Nitrogen (2.5.9)
AI blank absorbance of rite test solution; 2.4 per cent to 3.0 per cent (dried substance).
Az absorbance of the test solution with chondroitinase ABC;
A) absorbance of the test solution with chondroitinase AC; Nucleic acids
BI gradient of the curve obtained with the chondroitin sulfate Maximum 0.5 per cent (dried substance).
reference solutions with cbondroitinase ACj
Bz gradient of the curve obtained with the cbondroitin sulfate
Test solution Weigh about SO mg of the dried substance to
reference solutions with cbondroitinase ABC; be examined into a centrifuge tube and dissolve in 200 JlL of
8) gradient of the curve obtained with the dennatan sulfate waterR.
reference solutions with chondroiunase ABC;
Reference solution Dissolve about SO mg of ribonucleic
C concentration of the test solution, in milligrams per minilitrei
acidGRS in 5 mL of 0.1 M sodium hydroxide and dilute to
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1-718 Dantrolene Sodium 2022
20.0 mL with water R. Transfer 200 ~ of the solution into a bubbles to form. Add 40 llL of bovine factor Xa solution Rl to
centrifuge tube. eachwell. Exactly 2 min after the addition of the factor Xa
Add 4.0 mL of a 50 f!L solution of trichloroacetic acid R to solution, add 80 llL of chromogenic substrate R5. Measure the
each tube and mix. Place aU tubes in boiling water for absorbance at 405 nm (2.2.25) using a suitable reading
30 min. Allow to cool to room temperature. Add again device, exactly 4 min after the addition of the factor Xa
4.0 mL of a 50 f!L solution of trichloroacetic acid R to each solution. Measure the absorbance again at 405 nm (2.2.25),
tube and mix. If any of the test solutions is not clear, exactly 14 min after the addition of the factor Xa solution.
sonicate aU the tubes in an ultrasonic hath for 10 min and Use M (A,. - A,.) for the calculation of the anti-factor Xa
centrifuge at 1500 g for 15 min. Dilute 1.0 mLofthe clear activity. Determine the blank amidolytic activity in the same
supernatant to 4.0 mL with water R. Measure the manner, using tm(hydroxymeehyljaminomerhane BDTA buffer
absorbances of the diluted reference and test solutions at solution pH 8.4 R as the blank (minimum 8 blanks per
265 nm (2.2.25) against a blank solution prepared in the microtitre plate). Calculate the potency of the substance to
same manner, and calculate the percentage nucleic acid be examined in units of anti-factor Xa activity permilligram
content of the sample. usinga suitable statistical method, for example the parallel-
line assay (5.1).
Total protein (2.5.31, Method 2)
Maximum 0.5 per cent. STORAGE
Dissolve the substance to be examined in distilled water R. In an airtight container. If the substance is sterile, the
Use bovine albumin RJ as me reference substance. container is also sterile and tamper-evident.
Adjust the concentration of the diluted LABELLING
phosphomolybdorungstic reagent so that the pH in the The label states the number of units of anti-factor Xa activity
reaction mixture is between 10.0 and 10.5. per milligram.
Sodium _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PlJE<r

examined in 100.0 mL of a 1.27 mglmLsolution of caesium Dantrolene Sodium
chloride R in 0.1 M hydrochloric acid.
Reference solutions Prepare reference solutions containing
50 ppm, 100 ppm and 150 ppm of Na by diluting sodium NO.
standard solution (1000 ppm "Na) R with a 1.27 mg/mL
solution of caesium chloride R in 0.1 M hydrochloric acid.
Source Sodium hollow-cathode lamp.
Atomisation derm:e Air-acetylene flame.
399.3 24868-20-0
Maximum 5.0 per cent, determined on 0.500 g by drying in Action and use
an oven at 60°C over diphosphoms pentoxide' R at a pressure Skeletal muscle relaxant.
of 670 Pa for 3 h.
Preparation
Bacterial endotoxin. (2.6.14) Dantrolene Oral Suspension
Less than 0.02 ill per unit of anti-factor Xa activity, if
intended for use in the manufacture of parenteral DEFINITION
preparations without a further appropriate procedure for the Dantrolene Sodium is 1-(5-p-nitropheny1furfurylideneamino)
removal of bacterial endotoxins. hydantoin sodium. It contains not less than 98.0% and not
ASSAY more than 102.0% of C 1. H , N ..NaO" calculated with
Cany out the assay at room temperature. reference to the anhydrous substance.
The anticoagulant activity of danaparoid sodium is CHARACTERISTICS
determined in viero by an assaywhich determines its ability to A yellowish-orange to orange crystalline powder.
accelerate the inhibition of factor Xa by antithrombin ill Very slightly soluble in wa'er, slighdy soluble in ethanol
(anti-factor Xa assay). (96%); sparingly soluble in methanol; practically insoluble in
Test solutWnl Prepare 2 independent series of dilutions in acetone.
geometric progression of the substance to be examined in IDENTIFICATION
tris(hydroxymethyl)aminomethane BDTA buffer ",lution
A. The infrared absorption spectrum, Appendix Il AJ is
pH 8.4 R and in the concentration range of 0.1-0.32 units of concordant with the reference spectrum of dantrolene sodiwn
anti-factor Xa activity per millilitre.
(RS 422).
Reference solutions Prepare 2 independent series of dilutions
B. In the Assay, the chromatogram obtained withsolution
in geometric progression of danaparojd sodium CRS in
(1) shows a peakwith the same retention time as the
tris(hydroxymethyl)aminomethane BDTA buffer solutWn principal peak in the chromatogram obtained with
pH 8.4 R and in the concentration range of 0.08-0.35 units solution (2).
of anti-factor Xa activity per millilitre.
C. To 0.1 g of the substance being examined add 20 mL of
Transfer 40 J!l. of each solution into the wells of a 96-well
water and 2 drops of acetic acid, shake well and filter.
microtitre plate. Add 40 IlL of antithrombin lJl solution R4 to
The filtrate yields the reactions characteristic of sodium salts,
each well and shake the microtitre plate but do not allow
Appendix VI.
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2022 Damron 1-719
TESTS surface of which has been modified with chemically-bonded

A1kallnlty nitrile groups (Spherisorb CN is suitable).
Shake 0.7 g in 10 mL of water for 5 minutes and centrifuge. (b) Use isocratic elution and the mobile phase described
To 5 mL of the supernatant add 45 mL of water and 3 drops below.
of phe"clph,hakinsolurian R1 and 0.1 mL of 0.1M hydrochloric (c) Use a flow rate of I mL per minute.
acid VS. A red colour is not produced.
(d) Use an ambient column temperature.
Related substances (e) Use a detection wavelength of262 om.
Carry out the method for liquid chromatography,
Appendix ill D, using the foHowing solutions. (!) Inject 20 ~L of each solution.
(1) Dissolve 50 mg of the substance being examined in MOBILE PHASE

20 mL of tetrahydrofuran and 2 mL of glOOal acetic acid and 15 volumes of acetonitrile and 85 volumes of a phosphate
dilute with sufficient absolute ethanol to produce 100 mL. buffer pH 6.8 prepared by dissolving 11.88 g of disodium
(2) Dilute I mL of solution (I) to 100 mL with absolute hydrogen olthophosphate and 9.08 g of potassium dil!Ydrogen
ethanol. orthophosphate in 1000 mL of water.
(3) Dissolve 5 mg of damrolene sodium BPCRS and 0.1 g of DETERMINATION OF CONTENT
theopl!Ylline BPCRS in 20 mL of tetrahydrofuran and 2 mL of Calculate the content of Cl4.HgN~a05 in the substance
glacial acetic acid and dilute with sufficient absolute ethanol to being examined using the declared content OfC14HgN4Na05
produce 100 mL. Further dilute 10 mL of this solution to in dantrolene sodium BPCRS.
100 mL with absolute ethanol.
(a) Use a stainless steel column (15 em x 4.6 mm) packed
with silica gelfor chromatography (5 pm) (Zorbax Sil is Dantron
suitable).
(b) Use isocratic elution and the mobile phase described OH 0 OH
~
below.
(e) Adjust the flow rate of the mobilephase so mat the
retention time of the peak corresponding to Danrrolene
Sodium is about 8 minures. ~
o
(d) Use a column temperature of 30°.
(e) Use a detection wavelength 0000 om. 240.2 117-10-2
(!) Inject 10 ~L of each solution.
(g) For solution (I) allow the chromatography to proceed for Action and use
at least twice the retention time of the principal peak. Anthraquinone stimulant laxative.
MOBILE PHASE
Preparation
Co-danthrusate Capsules
9 volumes of absolute ethancl, 10 volumes of glacial acetic acid
and 90 volumes of hexane. DEFINITION
SYSTEM SUITABILITY Dantron is mainly l,8-dihydroxyanthraquinone. It contains
The test is not valid unless, in the chromatogram obtained not less than 98.0% and not more than 102.0% of total
with solution (3), the resolution between the peaks phenols, calculated as C l4Hs04 and with reference to the
corresponding to theophylline and danttolene is at least 6. dried substance.
LIMITS CHARACTERISTICS
In the chromatogram obtainedwith solution (1): An orange, crystalline powder.
the total area of aU the secondary peaks is not greater than the Practically insoluble in water; slightly soluble in ethe;, very
area of the principal peak in the chromatogram obtained with slightly soluble in ethanol (96%). It dissolves in solutionsof
solution (2) (1%). alkali hydroxides.
Water IDENllFICATION
14.5 to 17.0% w/w, Appendix IX C. Use 0.2 g A. The infrared absorption spearum, Appendix Il A, is
concordant with the reference spectrum of danrrcn (RS 083).
ASSAY
Carry out the method for liquidchrolTiatagrapl!Y, B. The ligh, absorption, Appendix II B, in the range 230 to
Appendix ill D, using the following solutions. 350 am of a 0.001% w/v solution in dichloromuhane exhibits
maximaat 255 nm and 285 nm and a less well-defined
maximwn at 275 run. The absorbance at the maximum at
50 mL of dimethylfonnamide and dilute I volume of the
255 nm is about 0.82 and at the maximum at 285 nm is
resulting solution to JOO volumes with the mobile phase.
about 0.48, each calculated with reference to the dried
(2) Dilute 1 volume of a 0.12% w/v solution of dantrolene substance.
sodium BPCRS in dimethylfonnamide to 100 volumes with the
C. Dissolve 5 mg in 5 mL of 1M sodium hydroxide. A clear
mobile phase. red solution is produced immediately.
TESTS
(a) Use a stainless steel column (15 cm x 4.6 mm) packed Mercury
with spherical particles of silica, 5 pm in diameter, the To 0.50 g in a Kjeldabl flask add 2.5 mL of "i'n, acid and
allow to stand until the initial vigorous reaction has subsided.
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1-720 Dapsone 2022
Add 2.5 mL of sul/un'c acid and heat until dense white fumes area of the principal peak in the chromatogram obtained
are evolved. Cool, add 2.5 mL of nitric acid and heat until with solution (2) (3.3% taking into account the correction
fumes are again evolved. Repeat the procedure with a further factor of the impurity);
2.5 mL of nitric acid, cool, add 50 mL of water, boil me - - the sum of the areas of any oilier secondary peaks is not
solution until the volume has been reduced to about 25 mL greater than the area of the principal peakin the
and cool. Transfer to a separating funnel using water, dilute chromatogram obtained with solution (2) (2%)j
to about 50 mL with water and add 50 mL of O.5M sulfuric - disregard any peak with a retention time less than one
acid. Add 100 mL of water, 2 g of hydroxylamine third of that of the principal peak.
hydrochloride, 1 mL OrO.OSM disodium edetate, 1 mL of glacial Loss on drying
acetic add and 5 mL of dichtoromahane, shake, allow to When dried to constant weightat 105°, loses not more than
separate and discard the dichloromethane layer. Titrate the 0.5% of its weight. Use 1 g.
aqueous layer with a 0.0008% wlv solution of dithizone in
dichloromethane, shaking vigorously after each addition, ASSAY
allowing the layers to separate and discarding Ute Dissolve 0.2 g in 50 mL of anhydrous pyridine and carry out
dichloromethane layer, until the dichloromethane layer Method II for non-aqueous titration, Appendix VDIA, using
remains green. Repeat the operation using a solution O.IM telralmtylammonium hydroxide VS as titrant and
prepared by diluting 1 mL of mercury standard solution determining the end point potentiometrically. Each mL of
(5 ppm Hg) to 100 mL with O.5M sulfuric acid and beginning 0.1M telrabucylammonium hydroxi'de VS is equivalent (0
at the words 'Add 100 mL of water... '. The volumeof the 24.02 mg of total phenols, calculated as C I4 H s0 4 .
dithizone solutionrequired by the substance being examined
does not exceed that required by the mercury standard
solution.
Related substances Dapsone
Carry out the method for liquidchromatography,
(Ph. Eur. managraph 0077)
Appendix ill D, using the following solutions.
20 mL of tarahydrcfuron and dilute to 100 mL with the
H,NY"u ONH,
mobile phase.
(2) Dilute I volume of solution (I) to 50 volumes with the
V o '" S
".~
0
mobile phase.
(3) Dissolve 50 mg of dantnm impurity standard BPCRS in 248.3 80-08-0
20 mL of tetrahydra/uran and dilute to 100 mL with the
mobile phase. Action and use
Folic acid synthesis inhibiter; treatment of leprosy.
Preparation
(a) Use a stainless steel column (25 em x 4.6 mm) packed
Dapsone Tablets
with oetadecylsily/ silica gelfar chromatagraphy (5 ~m)
(Nucleosil CI8 is suitable). PIIw _
(b) Use an isocratic system using the mobile phase described DEFINITION
below.
Dapsone containsnot less than 99.0 per cent and not more
(c) Use a flow rate of I mL per minute. than the equivalent of 101.0 per cent of
(d) Use an ambient column temperature. 4,4'-sulfonyldianiline, calculated with reference (0 the dried
(e) Use a detection wavelength of 254 nm. substance.
(f) Inject 20 ~L of each solution. CHARACTERS
(g) Allow the chromatography to proceed for 1.5 times the A white or slightly yellowish-white, crystalline powder, very
retention time of the principal peak. slightly soluble in water, freely soluble in acetone, sparingly
soluble in alcohol. J[ dissolves freely in dilutemineral acids.
MOBILE PHASE
A mixture of 2.5 volumes of glacial acetic acid, 40 volumes of IDENTIFICATION
tetrahydrofuran and 60 volumes of water. A. Meiring point (2.2.14): 175 °C to 181 °C.
SYSTEM SUITABILITY
B. Dissolve 50.0 mg in methanal R and dilute to 100.0 mL
with the same solvent. Dilute 1.0 mL of thissolution to
The test is not valid unless, in the chromatogram obtained
100.0 mL with methanol R. Examined between 230 om and
with solution (3):
350 om (2.2.25), the solution shows 2 absorption maxima, at
- - a peak due to I-hydroxyanthraquinone appears
260 om and 295 om. The specificabsorbances at these
immediately before the principal peak, as indicated in the
maxima are 700 to 760 and 1150 to 1250, respectively.
reference chromatogram supplied with dantron
;mpuniy standard BPCRSj C. Examine the chromatograms obtained in the test for
- the height of the trough separating the two peaks is not related substances. The principal spot in the chromatogram
greater than one third of the height of the peak due to obtained with test solution (b) is similar in position, colour
J-hydroxyanthraquinone. and size to the principal spot in the chromatogram obtained
LIMITS
TESTS
In the chromatogram obtained with solution (1):
- the area of any peak corresponding to Related substances
I-hydroxyanrbraqulnone is not greater than 2.5 times the Examine by thin-layer chromatography (2.2.27), using sitka
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2022 Daunorubicin Hydrochloride 1-721
Test solution (a) Dissolve 0.10 g of the substance to be Pl>E" _

examined in methanol R and dilute to 10 mL with the same DEFINITION
solvent. (8S, I 0S)-8-Acetyl-1 0- [(3-amino-2,3,6-trideoxy-«-t.-{yxo-
Testsolution (b) Dilute 1 mL of test solution (a) to 10 mL hexopyranosyI)oxy]-6,8, II-trihydroxy-l-methoxy-7,8, 9,10-
with methanol R. tetrahydrotetracene-5,12-dione hydrochloride.
Reference sohuion (a) Dissolve 10 mg of dapsone CRS in Substance produced by certain strains of Streptomyces
methanol R and dilute to 10 mL with the same solvent. coeruleornbidus or of Streptomyces peucetius or obtained by any
Reference solution (b) Dilute 1 mL of test solution (b) to othermeans.
10 mL with methanol R. Content
Reference solution (c) Dilute 2 mL of reference solution (b) 95.0 per cent to 102.0 per cent (arthydrous substance).
to 10 mL with methanol R.
PRODUCTION
Apply separately to the plate I ~L of test solution (b), I ~ It is produced by methods of manufacture designed to
of reference solution (a), 10 ~L of test solution (a), 10 ~L of eliminate or minimise the presence of histamine.
reference solution (b) and 10 I-IL of reference solution (c).
Develop in an unsaturated tankover a path of 15 em using a CHARAGrERS
mixture of 1 volume of concentrated ammonia R, 6 volumes of Appearance
methanol R, 20 volumes of ethylacetate Rand 20 volumes of Crystalline, orange-red powder, hygroscopic.
heptane R. Allow the plate to dry in air. Spray the plate with Solubility
a I gIL solution of 4-dimethylaminocinnamaldehyde R in a Freely soluble in water and in methanol, slightly soluble in
mixture of 1 volume of hydrochloric. acidRand 99 volumes of alcohol) practically insoluble in acetone.
alcohol R. Exantine in daylight. Any spot in the
IDENTIFICATION
chromatogram obtained with test solution (a). apart from the
principal spot, is not more intensethan the spot in the A. Infrared absorption spectrophotometry (2.2.24).
chromatogram obtained with reference solution (b) Comparison daunorubicin hydrochloride CRS.
(1.0 per cent) and not more than 2 such spots are more B. Dissolve about 10 mg in 0.5 mL of nitric acid R, add
intense than the spot in the chromatogram obtained with 0.5 mL of water R and heat over a flame for 2 min. Allow to
reference solution (c) (0.2 per cent). cool and add 0.5 mL of silvernitrate solution Rl. A white
Loss on drying (2.2.32) precipitate is formed.
Not more than 1.5 per cent, determined on 1.000 g by TESTS
drying in an oven at 10~ °C. pH (2.2.3)
Sulfated ash (2.4.14) 4.5 to 6.5.
Not more than 0.1 per cent, determined on 1.0 g. Dissolve 50 mg in carbon dioxide-free water R and dilute to
ASSAY J0 mL with the same solvent.
Dissolve 0.100 g in 50 mL of dilure hydrochloric acidR. Carry Related substances
out the determination of primary aromatic amino-nitrogen Liquid chromatography (2.2.29). Prepare the solutions
(2.5.8). immediately before use.
1 mL of 0.1 M sodium nitrite is equivalent to 12.42 mg of Test solution Dissolve 50.0 mg of the substance to be
C12H12N202S, examined in the mobile phase and dilute to 50.0 mL with
the mobile phase.
STORAGE
Store protected from light. Reference solution (a) Dissolve 50.0 mg of daunorubicin
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P/lElT
hydrochloride CRS in the mobile phase and dilute to 50.0 mL
Reference soluuon (b) Dissolve 10 mg of daxornbicin
hydrochloride CRS and 10 mg of epirnbicin hydrochloride CRS
in the mobile phase and dilute to 100.0 mL with the mobile
Daunorubicin Hydrochloride phase. Dilute 1.0 mL of the solutionto 10.0 mL with the
mobile phase.
Reference solution (c) Dissolve 5.0 mg of
o OH 0 daunornbicinone CRS and 5.0 mg of doxornbicin
~ I
hydrochloride CRS in the mobile phase and dilute (0
I "" ··OH
CH,
"" d' solution to 10.0 mL with the mobile phase.
H····.. , Hel
Reference solurion (d) Dilute 1.0 mL of reference solution (a)
OCH30 OH~O
CH, to 200.0 mL with the mobile phase.
HO Column:
NH, = =
- size: I 0.25 m, 0 4.0 mm,
- stationary phase: end-eapped oetadecy!si1Y1 silica gelfor
23541-5()'6
564.0
iWobile phase Mixture of equal volumes of acetonitrile R and
Action and use a solution containing2.88 gIL of sodium laurilsulfate R and
Cytostatic; anthracydine antibacterial. 2.25 gIL of phosphoric acidR.
Flow Tale 1 mUmin.
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1-722 Daunorubicin Hydrochloride 2022
Injection 5 J.lL; inject the test solution and reference o OH H OH

~CH'
~.
Run time Twice the retention time of daunorubicin.
Relative retention With reference [0 daunorubicin (retention un andepimeralC'
time = about 15 min): impurity A ;;;; about 0.4; OCH30 OH~"O
cli,
impurity D = about 0.5; epirubicin = about 0.6;
=
impurity B about 0.7. HO
NIi,
System suit{lbility Reference solution (b):
- resolution: minimum of 2.0 between the peaks due to
B. (8S, IOS)-I 0- [(3-amino-2,3,6-trideoxy-<.<-L-Iyxo-
impurity D and epirubicin.
hexopyranosyl)oxy]-6,8,II-trihydroxy-8-[(IRS)-I-
Limits: hydroxyethyl] -1-methoxy-7,8, 9,1 O-tetrahydrotetracene-
-impm;ty A: not more than the area of the corresponding 5,12-dione (daunorubicincl),
solution (c) (0.5 per cent), o OH
- impurity B: not more than 3 times the area of the
reference solution (d) (1.5 per cent), 4'
- impurity D: not more than the area of the corresponding
peak in the chromatogram obtained with reference o~H
0 "0
CH,
solution (c) (0.5 per cent),
HO
- any other impun'ty: not more than the area of the principal
NH,
solution (d) (0.5 per cent),
- total of other impurities: not more than 5 times the area of
c. (8S, IOS)-I 0- [(3-amino-2,3,6-trideoXY-<'<-L-1yxo-
hexopyranosyl)oxy]-6,8,II-trihydroxy-l-methoxy-8-(2-
the principal peak in the chromatogram obtainedwith
oxopropyl)-7,8,9, IO-tetrahydroterracene-S, 12-dione
reference solution (d) (2.5 per cent),
(feudomycin B),
the chromatogram obtained with reference solution (d) o OH 0
(0.05 per cent). OH
Butanol (2.4.24, System B)
Maximum 1.0 per cent. . 4'
H •.
Water (2.5.12) OH~O.O
Maximum 3,0 per cent, determined on 0.100 g. CIi,
Bacterial endotoxins (2.6.14) HO
Less than 4.3 IU/mg, if intended for use in the manufacture Nt<,
procedure for the removal of bacterial endotoxins. D. (8S,I OS)-I 0-[ (3-amino-2,3,6-trideoXY-<'<-L-(yxo-
hexopyranosyl)oxy]-6,8, II-trihydroxy-8-(bydroxyacetyl)-I-
ASSAY methoxy-7,8,9,1O-tetrahydrotetracene-5, 12-dione
Liquid chromatography (2.2.29) as described in the test for (doxorublcin),
related substances.
Injection Test solution and reference solution (a). o OH H OH
Calculate the percentage content of C27H30CINOIO'

STORAGE
~I
~I ~ " -, QH: CH,
un andepimeralC'
In an airtight container, protected from light. If the substance oc~o OHHOH

is sterile, store in a sterile, airtight, tamper-evident container,
IMPURITIES E. (8S, 10S)-6,?,1 0, II-tetrabydroxy-8-[(IRS)-I-
hydroxyethyl]-I-methoxy-7,8,9, IO-tetrabydrotetracene-
o OH 0 5,12-dione (13-dihydrodaunorubicinone),
~I
"'-
OCH30
'"
4'
OH H
\
"'OH CH,
OH
o OH 0
, CH,
4'
A. (8S, 1OS)-8-acetyl-6,8,1O,II-tetrabydroxy-I-methoxy- o~H

( ,"0
7,8,9, Io-tetrahydrotetracene-5, 12-dione (daunorubicin CH,
aglycone, daunorubicinone), HO
Nt<,
F. (8S,1 OS)-I 0-[(3-amino-2,3,6-trideoxy-<.<-L-1yxo-

bexopyranosyl) oxy]-6,8, II-trihydroxy-I-methoxy-8-
propanoyl-7,8,9,1O-tetrahydrotetracene-5,12-dione
(8-ethyldaunorubicin).
____ ~ PhE"
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2022 Deferasirox 1-723
Decyl Oleate Deferasirox

(ph. Eur. monograph 1307) (ph. Eur. monograph 2933)
Action and use
Excipient.
""'<r _
DEFINITION
Mixture consisting of decyl esters of fatty acids, mainly oleic
(cis-9-octadecenoic) acid.
A suitable antioxidant may be added.
CHARACTERS
Appearance 373.4 201531}-41-8
Clear, pale yellow or colourless liquid.
Solubility Action and use
Practically insoluble in water, miscible with ethanol Selective iron(IlI) chelator; treatment of iron overload
(96 per cent), with methylene chloride and with light f'llf<r _ _ ~ _
petroleum (bp: 40-60 "C).
DEFINITION
IDENTIFICATION
4- [3,5-Bis(2-hydroxypheny I) -I H- 1,2,4-triazol-I-yl] benzoic
A. Relative density (see Tests).
acid.
B. Saponification value (see Tests).
Content
C. Oleic acid (see Tests). 98.0 per cent to 102.0 per cent (anhydrous substance).
TESTS CHARACTERS
Relative density (2.2.5) Appearance
0.860 to 0.870. White or slightly yellow powder.
Acid value (2.5. I) Solubility
Maximwn 1.0, determined on 10.0 g. Practically insoluble in water, very soluble in dimethyl
IodIne value (2.5.4, MethodA) sulfoxide, slightlysoluble in anhydrous ethanol, practically
55 to 70. insoluble in heptane.
Peroxide value (2.5.5, MethodA) It shows polymorphism (5.9).
Maximum 10.0. IDENTIFICATION
Saponification value (2.5.6) Infrared absorption spectrophotometry (2.2.24).
130 to 140, determined on 2.0 g. Gomparison deferasirox GRS.
Oleic acid (2.4.22, MethodA)
TESTS
Minimum 60.0 per cent in the fatty acid fraction of the
Related substances
substance.
Liquid chromatography (2.2.29). Score the solutions at 2-8 "G.
Water (2.5.12)
Ruffer solution 0.100 gIL solution of sodium edetate R
adjusted to pH 2.1 with phosphork acid R.
Total ash (2.4./6) Solvent mixture 0.040 gIL solution of sodium edetate R,
Maximum 0.1 per cent, determined on 2.0 g. acetonitrile R (25:75 VIV).
STORAGE Testsolution (a) Dissolve 30.0 mg of the substance to be
Protected from light. examined in 15 mL of the solvent mixture with vigorous
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ "",<r shaking (this may take up to 20 min) and dilute to 20.0 mL
Test soluti<Jn (b) Dissolve 25.0 mg of the substance to be
the solventmixture.
Referenusolution (a) Dissolve 25.0 mg of deferasirox GRS in
mixture.
Reference solution (b) Dilute 1.0 mL of test solution (a) to
Reference solution (c) Dissolve 5 mg of deferasirox for system
suitability GRS (containing impurity D) in 8 mL of the
solventmixture with vigorous shaking and dilute to 10 mL
Reference sohuion (d) Dissolve 3.0 mg of deferasirox
impurity R CRS in the solvent mixture and dilute to 20.0 mL
with the solvent mixture. Dilure 5.0 mL of the solution to
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1-724 Deferasirox 2022
100.0 mL with the solvent mixture. Dilute 3.0 mL of this Reference solution (a) Dissolve 6.0 mg of dsferosirox
solution to 50.0 mL with the solvent mixture. impurity F CRS in I mL of dimethyl sulfoxide R and dilute to
Column: 20.0 mL with water R. Dilute 1.0 mL of the solution to
- size: 1 = 0.15 m, 0 = 3.0 mm; 10.0 mL with dimethylsulfoxide R.
- suuionatyphase: end-capped ocladecylsi(yl silica gef/or Reference solur;,m (b) Dilute 1.0 mL of reference solution (a)
chromowgraphy with embedded polargroups R (3.5 urn); to 10.0 mL with dimethyl sulfoxide R. To 1.0 mL of this
- temperature: 60 "C. solution add 5 mL of dimethyl sulfoxide Rand 20 mL of the
Mobile phase: solvent mixture. Heat this solution at 45 °C for 35 min, cool
- mobile phase A: acetonitrile R, buffer solution, waterfor to 2-8 "C and dilute to 50.0 mL with mobile phase A,
chromawgraphy R (10:10:80 VIVIV); previously cooled to 2-8 °C.
- mobile phase E: buffer solution, acetanitrile R (10:90 VIV); Reference solution (c) To 2.0 mL of reference solution (b)
add I mL of dimethylsulfoxide Rand 4 mL of the solvent
Tbn. Mobile phase A MobUe phase B mixture and dilute to 10.0 mL with mobile phase A.
(mIn) (per cent V/J? (pel' cent V/I1 Column:
0-10 62 38 - size: 1= 0.15 m, (2) = 3.0 mm;
10 - 14 62 ---> 0 38 -;0 100 - stationary phase: end-capped octadecylsi(yl silica gelfor
14 - 16 0 100 chromawgraphy R (3.5 pm);
Flow rate 0.8 mUmin. Mobile phase:
Detection Spectrophotometer at 250 nm. - mobIle phase A: phosphoric acid R, acetonitrile R, waterfor
Auueampler Set at 5 "C. chromatography R (2:100:900 VIVIV);
- mobile phase B: phosphoric acid R, waterfor
chromatography R, acetonitrile R (2:100:900 VIVIV);
solutions (b), (e) and (d).
Identification of impun"ties Use the chromatogram supplied Time Mobile phase A MobUe phase B
with deferasirox for system suitabIlity CRS and the (min) (per cent V/V.) (per cent V/V.)
chromatogram obtained with reference solution (c) to identify 0-2 90 10
the peak due to impurity D; use the chromatogram obtained 2-8 90 -+ 58 10 ..... 42
with reference solution (d) to identify the peak due to 8 - 8.1 58 ..... 0 42 -+ 100
impurity B. 8.1 - 16 0 100
Relative retention With reference to deferasirox (retention
=
time about 10 min): impurity B about 0.5; = F/ow rate 1.0 mUmin.
=
System suitability:
Injection 25 J1L of the test solutionand reference
impurity D and deferasirox in the chromatogram obtained
with reference solution (c)j Relative retention With reference to deferasirox (retention
- signal-to-noise ratio: minimum 10 for the principal peak in time = about 10 min): impurity F acetone
the chromatogram obtained with reference solution (d). derivative = about 0.5.
Cakulatian of percentage contents:· System suitability:
- for each impurity, use the concentration of deferasirox in - signal-to-noise ratio: minimum 10 for the peak due to
reference solution (b). impurity F acetone derivative in the chromatogram
obtained with reference solution (c);
Limits:
- rejmltability: maximum relative standard deviation of
5.0 per cent for the peak. due to impurity F acetone
0.05 per cent;
derivative determined on 6 injections of reference
- IOtal: maximum 0.2 per cent;
solution (b).
Calculation of percentage contear:
Impurity F
- for impurity F, use the concentration of impurity F in
liquid chromatography (2.2.29). Protect the solutionsjrom
light:
Limit:
Solvent mixture phosphoric acidR, waterR, aatone R
- impurity F: maximum 0.5 ppm.
(25:100:900 VIVIV).
Water (2.5.12)
Teu solution Dissolve 0.600 g of the substance to be
examined in I mL of dimethyl sulfoxide R, add 2 mL of the
solvent mixture and mix thoroughly using a vortex mixer. Sulfated ash (2.4.14)
Heat the solution at 45 °C for 35 min, then cool to 2-8 °C Maximum 0.1 per cent, determined on 1.0 g.
for 1 h. Dilute to 5.0 mL with mobile phase A, previously ASSAY
cooled to 2-8 °C. Shakevigorously usinga mechanical shaker Liquid chromatography (2.2.29) as described in the test for
for 2 min. Centrifuge immediately at 4000 g for 5 min and related substances with the following modification.
filter the supernatant through a membrane filter (nominal
pore size 0.45 urn). If a precipitation is still observed, allow
to stand for 1 h at 2-8 °C. Filteragain through a membrane Calculate the percentage content of C21H15N304 taking into
filter (nominal pore size 0.45 pm) immediately before account the assigned content of deferasirox CRS.
injection.
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2022 Deferiprone 1-725
IMPURITIES
Specified impurities F.
Other detectable impurities (thejolb>wing substances would, If
present at a sufficient level) be delated by one or other of the tests
in the monograph. They are limited by the general acceptance F. d-hydrezinylbenzoic acid.
ctitenon for other/unspecified impurities andlor by thegeneral _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I'IIEII
there/ore not n«essary to identify these impurities for
demonstration of compliance. See also5.10. Contrd of impurities
in substances for pharmaceutical use) A J BJ C, D, E.
Deferiprone
A. 2-hydroxy-N-(2-hydroxybenzoyl)benzamide,
~r
G,H,NO, 139.2 30652-//-0
OH
UoY-J.-, Action and use
Chelating agent (iron).
U Preparations
Deferiprone Oral Solution
B. 2-(2-hydroxyphenyl)-4H-I,3-benzoxazin-4-one, Deferiprone Tablets
PIIEII _
DEFINITION
3-Hydroxy-I,2-dirnethylpyridin-4-(IH)-one.
Content
CHARACTERS
Appearance
White or pinkish-white powder.
C. 2- [3,S-bis(2-hydroxyphenyl)-IH-1,2,4-triazol-I-yl] benzoic Solubility
acid, Sparingly soluble in water, slightly soluble in anhydrous
IDENTIFICATION
Comparison d'fen'prone CRS.
TESTS
Related substances
Liquid chromatography (2.2.29). Use onlycdoudess glassware.
Protect the solutions from lighl.
Solution A Dissolve 2.91 g of sodium edetate R, 4.01 g of
D. 3-[3,S-bis(2-hydroxyphenyl)-1 H-I ,2,4-triazol-I-yl]benzoic sodium oaanesulfonau monohydrate R and 6.20 g of dipotassium
acid, hydrogen phosphate R in waterfor chromatography R and dilute
to 2000 mL with the same solvent; adjust to pH 3.0 with
phosphoric acidR.
Solution B Dissolve 0.73 g of sodium edetou R, 1.0 g of
sodium oeumesulfonate monohydrate Rand 1.55 g of dipotassium
hydrogen phosphaw R in Waler for chromatography R and dilute
to 2000 mL with the same solvent; adjust to pH 3.0 with
phosphoric acid R.
Solvent mixture acetonitrile R, water R (10:90 VIP).
examined in a volume of me mobile phasecorresponding to
E. ethyl 4-[3,S-bis(2-hydroxyphenyl)-IH-I,2,4-triazol-I-yl] about 2/3 of the final volume and dilute to 100.0 mL with
benzoate, the mobile phase.
Testsolution (b) Dissolve 50.0 mg of the substance to be
examined in a volume of the solventmixture corresponding
to about 2/3 of the final volume and dilute to 50.0 mL with
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1-726 Demeclocycline Hydrochloride 2022
the solvent mixture. Dilute 5.0 mL of the solution to Otherdetectable impunues (thefollowing substances would, if
200.0 mL with the mobile phase. present at a sufficient level) be detected by oneor other of the USts
Reference solution (a) Dissolve 2 mg of maltolR (impurity B) in the monograph. They are limited by the general acceptance
in the mobile phase and dilute to 100.0 mL with the mobile criterion for other/unspecified impuruies. It is therefore not
phase. To 2.5 mL of the solution add 10 mL of test necessary to identify these impurities for demonstration of
solution (a) and dilute to 100.0 mL with the mobile phase. compliance. See also5.10. Control of impurities in substances for
phamraceutical use) A.. C.
Dilute 1.0 mL oftest solution (a) to
Reference solutio" (1))
Reference solutio" (c) Dissolve 50.0 mg of deferiprone CRS in
a volumeof the solvent mixture corresponding to about 2/3
of the final volumeand dilute to 50.0 mL with the solvent
mixture. Dilute 5.0 mL of the solution to 200.0 mL with the A. l-methyl-3-(methylamino)-I,5-dihyclro-2H-pyrrol-2-one,
mobile phase.
Column:
- size: 1= 0.15 rn, 0;;;; 4.6 mm;
- stationary phase: styrene-diuinylbenzene copolymer R (5 JIm),;
Mobile phase aceto"ilTile R, solution A (10:90 VW). B. 3-hyclroxy-2-methyl-4H-pymn-4-one (malrol),
Preconditioning of the column Rinse for 20 min with the
mobile phase before each series of injections.
Injution 20 JlL of test solution (a) and reference
solutions (a) and (b). C. I ,2-dimethyl-4-[(8) -methylimino]-1,4-dihydropyridin-3-01.
Run time 3.5 times the retention time of deferiprone. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Identification of impun'ties Use the chromatogram obtained
impurity B.
Relativeretention With reference to deferiprone (retention Demeclocycline Hydrochloride
time =about 12 min): impurity B =about 0.5.
System suitability Reference solution (a): (Ph. Eur. monograph 0176)
~
OH 0 OHOHO 0
impurity B and deferiprone.
Calculation of percentage contents: <? NH,
- for each impurity, use me concentration of deferiprone in ""IH H I .HCI
. . OH
reference solution (b). Cl He) H H rN-CH3
Limits: H,C
- unspecified impurities: for each impurity, maximwn 501.3 64-73-3
0.05 per cent;
- total: maximum 0.2 per cent; Action and use
- reporting threshold: 0.03 per cent. Tetracycline antibacterial.
Water (2.5.32) Preparation
Maximum 0.5 per cent, determined on 0.100 g by direct Demeclocycline Capsules
sample introduction.
PhE" _
Sulfated ash (2.4. H)
Maximum 0.1 per cent, determined on 1.0 g. DEFINTIlON
(4S,4aS,5aS,6S,12aS)-7-CWoro-4-(dimethylamino)-
ASSAY
3,6)I0, 12)12a-pentahydroxy-I,11-dioxo-I,4)4a)5,5a,6) 11,12a-
octahydrotetracene-2-carboxamide hydrochloride.
related substanceswith the following modifications.
Substance produced by certain strains of Streptomyces
Mobile phase acetonitrile R, solution B (10:90 VIV).
aureofaciens.
Injection 10 IJL of test solution (b) and reference
solution (c). Content
89.5 per cent to 102"0 per cent (anhydrous substance).
Run time Twice the retention time of deferiprone.
Retention time Deferiprone = about 7.7 min. CHARACTERS
Appearance
Calculate the percentage content of C7HgN02 taking into
Yellow powder.
account the assigned content of dejen"prone CRS.
Solubility
IMPURITffiS Soluble or sparingly soluble in water, slightly soluble in
Specified impurities B. alcohol, very slightly soluble in acetone. It dissolves in
solutions of alkali hydroxides and carbonates.
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2022 Demeclocycline Hydrochloride 1-727
IDENTIFICATION Mobile phase:

A. Thin-layer chromatography (2.2.27). - mobile phaseA: acetonitrile R, waterfor chromatography R,
Test solution Dissolve 5 mg of the substance to be examined buffer I, buffer 2 (2:28:35:35 VIVIVIV);
in methanolR and dilute to 10 mL with the same solvent. - mobile phase B: acetonitrile R, buffer 1, buffer 2
(30:35:35 VIVIV);
Reference solution (a) Dissolve 5 mg of demedocydine
Tho. Mobile phase A Mobile phase B
same solvent. (min) (per cent V/JI) (per cent VIJI)
Reference solution (b) Dissolve 5 mg of demedocydine 0-5 83 17
hydrochloride CRS, 5 mg of chlortetracycline hydrochloride Rand 5 - 15 83 -+ 30 17 --> 70
5 mg of tetracydine hydrochloride R in methanol R and dilute to 15·25 30 70
J 0 mL with the same solvent.
Plate nc octadecylsi/yl silica gel Fm plateR. Flow rate I mIlrnin.
Mobile phase Mix 20 volumes of acetonirrile R, 20 volumes Detection Spectrophotometer at 280 run.
of methanol Rand 60 volumes of a 63 gIL solution of oxalic
acidR previously adjusted to pH 2 with concentrated
ammonia R.
Application I ur,
with demeclocydme for system suitability CRS and the
Development Over 3/4 of the plate. chromatogram obtained with reference solution (e) to identify
Drying In air. the peaks due to impurities A, B, C, E and G.
Detection Examine in ultraviolet light at 254 nm. Relativeretention With reference to demeclocycline
Sysrem suitability The chromatogram obtained with =
(retention time about 14 min): impurity C ;:: about 0.3;
reference solution (b) shows 3 clearly separated spots. =
impurity B about 0.7; impurity A ;:: about 0.8;
Results The principal spot in the chromatogram obtained =
impurity E about 1.2; impurity G about 1.6.=
with the test solution is similar in position and size to the System suirability Reference solution (c):
principal spot in the chromatogram obtained with reference - resolution: minimum 2.5 between the peaks due to
solution (a). impurities A and B.
B. To about 2 mg add 5 mL of sulfuric acidR. A violet Calculation of percctltage contents:
colour develops. Add the solution to 2.5 mL of water R. - for each impurity, use the concentration of
The colour becomes yeHow. demeclocycline hydrochloride in reference solution (b).
C. It gives reaction (a) of chlorides (2.3.1). Limits;
- impurities A, B: for each impurity, maximum 5.0 per cent;
TESTS
- impurities C, G: for each impurity, maximum 0.3 per cent;
pH (2.2.3)
- impun'ty E: maximwn 0.2 per cent;
2.0103.0.
- any otherimpurity: for each impurity, maximum
Dissolve 0.1 g in carbon dioxide-free waterR and dilute to 0.15 per cent;
10 mL with the same solvent. - total: maximwn 10.0 per cent;
Related substances - reporting threshold: 0.05 per cent.
Liquid chromatography (2.2.29). Prepare the solutions Water (2.5.1Z)
immediately before use. Maximum 3.0 per cent, determined on 1.000 g.
Buffer 1 22.2 gIL solution of sodium edetote R adjusted to Sulfated ash (2.4.14)
pH 7.5 with a 40 gIL solution of sodium hydroxitk R. Maximum 0.5 per cent, determined on 1.0 g.
Buffer 2 17.0 gIL solution of tetrapropylammonium hydrogen
ASSAY
sulfate R adjusted to pH 7.5 with a 40 gIL solution of sodium
hydroxide R.
examined in a 1.0 gIL solution of hydrochloric acid Rand
Injection 10 J.1L of the test solution and reference
solution (a).
dilute to 50.0 mL with the same solution.
Reference solution (a) Dissolve 25.0 mg of demedocycline Calculate the percentage content of C2IH22CIzN20s using
the chromatogram obtained with reference solution (a) and
hydrochloride CRS in a 1.0 gIL solution of hydrochlcri< acidR
taking into account the assigned content of demedocydine
and dilute to 50.0 mL with the same solution.
hydrochloride CRS.
100.0 mL with a 1.0 gIL solution of hydrochloric acidR. STORAGE
Reference solution (c) Dissolve 5 mg of demedocydine for Protected from light.
system suitability CRS (containing impurities A, B, C, E IMPURITIES
and G) in a 1.0 gIL solution of hydrochloric acidR and dilute Specified impuruies A, B, C, E, G.
to 10 mL with the same solution. Other detectable impurities (thefollowing substances. would, .]
Column: present at a sufficient level, be detected by one orotherof the tests
=
- size: 1= 0.075 m, 0 4.6 rom; in the monograph. They are limited by the general acceptance
- stationary phase: end-capped oaylsilyl silica gelfor criterion for other/unspecified impuniies and/or by the general
chromawgraphy Wlih embedded polar groups R (3.5 um); monograph Substances for pharmaceutical use (2034). It is
- temperature: 40 "C. therefore not necessary 10 identify these impurities for
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1-728 Deptropine Citrate 2022
demonstration of compliance. See also5.10. Control 0/impurities

in subsumus forpharmaceutical use) D, F.
F. (4R,4aS,12aS)-7-chIoro-4-(dimethylamino)-3,1O,11,12a-
tettahydroxy-l,12-dioxo-l,4,4a,5)12,12a-
hexahydrotettacene-2-carboxamide
(4-epianhydrodemeclocycline),
A. (4S,4aS,5aS,6S,12aS)-4-(dimethylamino)-3,6,1O,12,12a-
pentahydroxy-I, Il-dioxo-l,4,4a,5,Sa,6, 11, 12a-
octahydrotetracene-2-earboxamide (demethyltetracycline),
G. (4S,4aS,12aS)-7-chIoro-4-(dimethylamino)-3,10,II,12a-
tetrahydroxy-l , J2-dioxo-J,4,43,5,12,123-
hexahydrotetracene-2-carboxamide
B. (4R,4aS,5aS,6S,12aS)-7-chIoro-4-(dimethylamino)- (anhydrodemeclocycline).
3,6,10, 12,12a-pentahydroxy-l J l l-dioxo- _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
1,4,43,5,5a,6, 11,12a-octahydrotetracene-2-carboxamide
(4-epidemeclocycline),
Deptropine Citrate ***

** **
(Ph. Bur. monograph 1308) *****
C. (4R,4aS,5aS,6S, 12aS)-4-(dimethylamino)-3,6, 10,12,12a-

pentahydroxy-l,ll-dioxo-l ,4,4a,5,59,6,11, 12a-
octahydrotetracene-z-carboxamide
(4-epidemethyllelracycline),
C"H,,NO. 525.6 2169-75-7
Action and use

Histamine HI receptor antagonist; anticholinergic.
PhE" _
DEFINlTION
Deptropine citrate containsnot less than 98.0 percent and
D. (4R,4aS,12aS)-4-(dimethylamino)-3,10,11,12a- not more than the equivalent of 101.0 per cent of
tetrahydroxy-I J 12-dioxo-l,4,4a,S, 12,12a- ( 1R,3r,5S) -3-( 10, ll-dihydro- 5H-dibenzo[a,d] [7]annulen-5-
hexahydrotetracene-2-carboxamide yloxy)-8-methyl-8-azabicydo[3.2.I]octane dihydrogen citrate,
(4-epianhydrodemethyltetracycline), calculated with reference to the dried substance.
CHARACTERS
A white or almost white, microcrystalline powder, very
slightly soluble in water and in ethanol, practically insoluble
in methylene chloride.
It melts at about 170°C, with decomposition.
IDENTIFICATION
E. (4S,4aS, 12aS)-4-(dimethylamino)-3,10,II,12a-
Fint identification: A.
tettahydroxy-lJ 12-dioxo-l,4,43,5,12, 12a- Second identification: B, C, D, E.
hexahydrotettacene-2-carboxamide A. Examine by infrared absorption spectrophotometry
(anhydrodemethyltetracycline), (2.2.24), comparing with the spectrum obtained with
deptropine citrare CRS.
related substances. The principal spot in the chromatogram
with reference solution (b).
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2022 Dequalinium Chloride 1-729
C. To about I mg add 0.5 mL of sulfuric acid R. A stable I mL of 0.1 M perchloric acid is equivalent to 52.56 mg of
red-orange colour develops. C29H3SNOs·
D. Dissolve about 1 mg in 0.25 mL of perch/one acid Rand STORAGE
warm gently until the solution becomes turbid. Add 5 mL of Store protected from light.
gladal acetic add Rj a pink colour with an intense green
fluorescence appears. IMPURITIES
E. To about 5 mg add I mL of acetic anhydride Rand 5 mL
of pyridine R. A purple colour develops.
TESTS
pH (2.2.3)
Suspend 0.25 g in carbon dioxide-free water R, dilute to 25 mL
with the same solvent and filter. The pH of the solution is
3.1 to 4.5. A. (IR,3r,5S)-8-methyl-8-azabicydo[3.2.![octan-S-ol
(tropine),
Related substances
Examine by thin-layer chromatography (2.2.27), using as the
coating substance a suitable silica gel with a fluorescent l H
~I---t,-~~-~-~;]
indicator having an optimal intensity at 254 nm.
examined in methanol R and dilute to 10 mL with the same ~ .
solvent. H
with methanol R. B. (IR,3s,5S)-3-(1 0,II-clihydro-5H-dibenzo[a,d] [1]annulen-
Reference solution (a) Dilute 1.0 mL of test solution (a) to 5-yloxy)-8-methyl-8-azabicyclo[3.2.1]octane
100.0 mL with methanol R. (pseudodeptropine),
Reference solution (b) Dissolve 20 mg of deptropine
citrate CRS in methanol R and dilute to 2 mL with the same
solvent. Dilute 1 mL of the solution to 10 mL with
methanol R.
Reference solution rc) Djssolve 5 mg of tropine CRS in
methanol R and dilute to 100.0 mL with the same solvent.
Reference solution (d) Dissolve 10 mg of deptropine
citra te CRS and 10 mg of tropine CRS in methanol Rand
dilute to 25 mL with the same solvent. C. I 0,II-dihydro-5H-dibenzo[a,d] [1]annuien-5-01
(dibenzocycloheptadienol),
Apply to the plate 40 IlL of each solution. Develop over a
path of 10 em using a mixture of 8 volumes of concentrated
ammonia Rand 92 volumes of butanolR. Dry the plate at
100°C to 105°C until the ammonia has completely
evaporated. Examine in ultraviolet light at 254 nm. Any spot
in the chromatogram obtained with test solution (a), apart
from the principal spot, is not more intense than the spot in
(I per cent). Spray with dilute potassium iodobismutha..
solution R and then with a 10 gIL solution of sodium mirite R. D. (IR,3r,5S)-3-(IO,II-clihydro-5H-dibenzo[a,d] [1]annulen-
Expose the plate to iodine vapours. Examine in daylight and 5-yloxy)-8-azabicyclo[3.2.I]octane (demethyldeptropine).
in ultraviolet light at 254 nm. In the chromatogram obtained _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ""E<I
with test solution (a): any spot corresponding to tropine is
not more intense than the spot in the chromatogram
obtained with reference solution (c) (0.5 per cent); any spot,
apart from the principal spot and any spot corresponding to Dequalinium Chloride ***
tropine, is not more intense than the spot in the *** ***
chromatogram obtained with reference solution (a) (Ph. Eur. monograph 1413) ***
(I per cent). The test is not valid unless the chromatogram
obtained with reference solution (d) shows two clearly
separated spots.
Loss nn drying (2.2.32)
ASSAY
521.6 522-51-0
Dissolve 0.400 g in 50 mL of anhydrous acetic acidR. Titrate
with 0.1 lH perchloric acid, determining the end-point Action and use
potentiometrically (2.2.20). Antiseptic.
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1-730 Dequalinium Chloride 2022
PhEIr _ Column:
- size: 1 = 0.25 m, 0 = 4.6 mm;
DEFINlTION
- ,tationary phase: end-capped octade<y/silyl silica gelfor
I, I '-(Decane-I, I O-diyI)bis( 4-amino-2-methylquinolinium)
chromarography R.
dichloride (dried substance).
MobIle phase Dissolve 2 g of sodium hexanestdfonate R in
Content 300 mL of waterR; adjust to pH 4.0 with acetic acidRand
add 700 mL of methanol R.
CHARACfERS Floro rate 1.5 mUmin.
Appearance Deuaion Spectrophotometer at 240 DOl.
White or yellowish-white powder, hygroscopic.
lnjettion I0 ~L
Solubility
Run time 5 times the retention time of dequalinium
chloride.
IDENTIFICATION System suitability Reference solution (a):
First identification: B, E. - peak-to-1Jalley ratio: minimum 2.0, where Hp = height
Second identification: A, C, D, E. above the baseline of the peak due to impurity Band
A. Ultraviolet and visible absorption spectrophotometry HfJ = height above the baselineof the lowest point of the
(2.2.25). curve separating this peak from the peak due to
dequalinium chloride. If necessary, adjust the
Testsolution Dissolve about lOrng in water R and dilute to
concentration of methanol in the mobile phase.
100 mL with the same solvent. Dilute 10 mL of the solution
to 100 mL with waterR. Limits:
Spectral range 230-350 nm.
- impurity A: not more than 0.5 times the area of the
Absorption maxima At 240 nm and 326 nm. reference solution (b) (l per cent);
Shoulder At 336 nm. - totalof impun·ties other than A: not more than 5 times the
Absorbance ratios: area of the principal peak in the chromatogram obtained
- A2..JA32 6 =1.56 to 1.80; with reference solution (b) (10 per cent);
- A3201A,,. =1.12 to 1.30. - disregard limit: 0.025 times me area of the principal peak
B. Infrared absorption spectrophotometry (2.2.24). in the chromatogram obtainedwith reference solution (b)
(0.05 per cent).
Spectral range 600-2000 cm-I .
Comparison dequaJinium chl~ride CRS. Readily carbonisable substances
Dissolve 20 mg in 2 mL of "'!turi< acidR. After 5 min the
C. To 5 mL of solution S (see Tests) add 5 mL of potassium
solution is not more intensely coloured than reference
fern'cyanide solution R. A yellow precipitate is formed, solution BY. (2.2.2, Method I).
D. To 10 mL of solution S add I mL of dilute nitric acidR.
A white precipitate is formed. Filterand reserve the filtrate
Maximum 7.0 per cent, determined on 1.000 g by drying at
for identification test E.
105 °C at a pressure not exceeding 0.7 kPa.
E. The filtrate from identification test D gives reaction (a) of
chlorides (2.3.1). Sulfated ash (2.4.14)
TESTS
Solution S
ASSAY
Dissolve 0.2 g in 90 mL of carbon dioxide-free waterR,
In order w aooid ooerheating in the reaaion medium, mix
heating if necessary, and dilute to 100 mL with the same thoroughly throughou« and ,top the titration immediately afterthe
solvent. end-point has been reached.
Dissolve 0.200 g in 5 mL of anhydrous formic acidR and add
50 mL of acetic anhydride R. Titrate with O. I M perchwri<
acid, determining the end-point potentiometrically (2.2.20).
Acidity or alkalinity 1 mL of 0.1 M perchloric acid is equivalent to 26.38 mg of
C,oH.oCI 2 N. . .
,oIulion Rt. Not more man 0.2 mL of 0.01 M hydrochwri<
acid or 0.01 M sodium hydroxide is required to change the STORAGE
colour of the indicator. In an airtight container.
Liquid chromatography (2.2.29). Specified impurities A.
Test solution Dissolve 10.0 mg of the substance to be Otherdetectable impurities (thefollowing ",b,tances would, if
examined in the mobile phase and dilute to 10.0 mL with preunt ar a sufficient level, be deuaed by one or other of the tests
the mobile phase. in the monograph. They are limited by thegeneral acceptance
Reference solution (a) Dissolve 10.0 mg of dequalinium critetion for other/unspecified impurities and/or by thegeneral
chloride for petformance testCRS in the mobile phase and monograph Substances for pharmaceutical we (2034). It is
dilute to 10.0 mL with the mobile phase. therefore not ntXessary to identify these impun'ties for
Reference sdtuion (b) Dissolve 10.0 mg of dequalinium demonstration of compliance. See also 5.10. Control of impurities
chloride CRS in me mobile phase and dilute to 10.0 mL with in substances for pharmaceutical use) B, C.
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2022 3-0-Desacyl-4'-Monophosphoryl Lipid A 1-731
H'N~,rI
the particula r vaccine of proven clinical efficacy and safety in
man.
[ N During development studies, and wherever revalidation is
§
necessary, a test for residual endotoxin activity is carried out
CH, by injecting intravenously 12-day-old embryonated hens' eggs
with 0.1 mL of dilutions of the test sample (8 eggs per
A. 2-methylquinolin-4-amine, dilution) of 3-0-desacyl-4'-monophosphoryllipid A. Eggs are
candled and read for mortality at 18-24 hours post-
inoculation and the chick embryo 50 per cent lethal dose
(CELDso) is calculated. The residual endotoxin activity of
the 3-0-desacyl-4'-monophosphoryllipid A is acceptable if
the CELD,o is more than 100 pg.
An endotoxin standard of Salmonella typhimurium is prepared
and selected dilutions are injected into each group of 8 eggs.
For a test to be valid, the CELDso of the endotoxin standard
B. 4-amino-I-[10-[(2-methylquinolin-4-yl)amino]decyl]-2-
must not be more than 0.05 pg.
methylquinolinium chloride,
Reference preparation A batch of 3-0-desacyl-4'-
acr monophosphoryl lipid A shown to be comparable in structure
and function with a preparation of 3-0·desacyl-4'-
monophosphoryllipid A used as adjuvant in the particular
vaccine of proven clinical efficacy and safety in man or a
batch representative thereof.
BACTERIAL SEED LOTS
The bacterial strain used for master seed lots shall be
identified by historical records that include information on its
origin and the tests used [Q characterise the strain, in
C. 1-[10-(4-amino-2-methylquinolinio)decyl]-4-[[10-(4- particular genotypic and phenotypic information. Only a
amino-2-methylquinolinio)decyl]amino]-2- working seed lot that complies with the following
methylquinolinium trichloride. requirements may be used.
_ _ _ _ _ _ _ _ _ _- - - - - - - - - - ~PhE,;
Identification
The working seed lot is identified by suitable methods such
as Gram staining and fatty acid profiling (5.J.6).
Microbial Purity
3-D-Desacyl-4'·Monophosphoryl Each seed lot complies with the requirements for absence of
Lipid A contaminating organisms. Purity of bacterial cultures is
verified by methods of suitable sensitivity and specificity.
PROPAGATION AND HARVEST
PhE,; _
The bacteria is grown using a suitable liquid medium. At the
DEFINITION end of cultivation) the culture is tested for purity and yield.
3-0-Desacyl-4'-monophosphoryllipid A is a detoxified The culture medium is separated from the bacterial mass by
derivative of the lipopolysaccharide (LPS) of Salmonella a suitable method, for example filtration. Only a harvest that
minnesota, strain R595, which retains the immunostimuJatory is consistent with respect [Q the profiles for growth rate, pl-l,
activities of the parent LPS. It consists of a mixture of and 02-consumption may be used for the extraction of LPS.
congeners, all containing a backbone of III ,--+6-linked TRIETHYlAMINE SALT OF 3-0-DESACYL-4'-
disaccharide of 2-deoxy-2-aminoglucose phosphorylated at MONOPHOSPHORYL LIPID A
the 4 f -position, but differing in the fatty acid substitutions at LPS is extracted from the bacterial cells by successive alcohol
the 2, 2' and 3/ positions. The immunostimuJatory activities and chloroform-methanol extractions and is then converted
of 3-0-desacyl-4'-monophosphoryllipid A combined with to 3-D-desacyl-4'-monophosphoryllipid A by hydrolysis,
the vaccine include up-regulation of co-stimulatory molecules then purified and salified by triethanolamine before freeze-
on antigen-presenting cells and secretion of pro-inflammatory drying. The freeze-dried triethylamine salt of 3-D-desacyl-4'-
cytokines, resulting in an enhanced immune response of the monophosphoryl lipid A must comply with the following
Thl-type against the antigens. 3-D-desacyl-4'- requirements.
monophosphoryl lipid A is a lyophilised powder or a sterile Appearance
liquid. A visual description of the particular preparation after freeze-
Requirements given in the sections up to and including the drying is established and approved by the competent
section Triethylamine salt of3-G-desacyl-4'-monophosphoryl authority; each batch of freeze-dried triethylamine salt of
lipid A also apply to formulations that do not proceed to the 3-0-desacyl-4'-monophosphoryllipid A must comply with
3-D-desacyl-4'-monophosphoryl lipid A liquid bulk. this description.
PRODUCTION Protein
GENERAL PROVISIONS Less than 0.5 per cent mlm, determined using a suitable
The production method shall have been shown to yield method, for example a reversed-phase HPLC method for
consistently a 3-0-desacyl-4'-monophosphoryllipid A amino acid analysis (2.2.56). The total amino acid content in
comparable in structure and function with a preparation of micrograms is calculated by comparison to amino acid
3-G-desacyl-4'-monophosphoryl Iipid A used as adjuvant in standards and is equal to the protein concentration.
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1-732 3-0-Desacyl-4'-Monophosphoryl Lipid A 2022
Nucleic acld Identification) Tests and Assay and that is within the limits
Maximum 0.3 per cent 11'1111'1, determined using a suitable approved for the particular product may be used for the
method. For example, a ftuorimetric method may be used preparation of 3-0-desacyl-4'-monophosphoryl lipid A in the
where nucleic acids are extracted from the freeze-dried final lots.
triethylamine salt of 3-0-desacyl-4'-monophosphoryllipid A, CHARACTERS
using a solution containing NlltOH and a suitable non-ionic
When dispersed in an aqueous solution: slightly turbid
detergent, and stained by a suitable fluorescent dye.
suspension.
The nucleic acid content in the test sample is interpolated
from a calibration curve. When dissolved in an organic solvent: a description of its
appearance is established and approved by the competent
Hexosamine (2.5.21J) authority; the 3-0-desacyl-4'-monophosphoryllipid A liquid
1000 nmol/mg to 1450 nmol/mg. bulk complies with this description.
Phosphorus (2.5.11!)
IDENTIFICATION
0.5 umolrmg to 0.8 umol'mg.
Congener distribution (see Tests).
Congener distribution
The relative amount of tetraacyl, pentaacyl, hexaacyl and TESTS
heptaacyl congener groups are determined by a suitable Particle size
method, for example reversed-phase HPLC analysis (2.2.29). Where applicable) the particle size in the microfluidised
liquid bulk is determined by a suitable method) for example
The relative amount of each congener group in the
dynamic light scattering. The particle size for each batch of
triethylamine salt of3-0-desacyl-4'-monophosphoryllipid A
liquid bulk is within the limits approved for the particular
is:
- tetraacyl: 15 per cent to 35 per cent; product.
- pentaacyl: 35 per cent to 60 per cent; Sterility (2.6.1)
- hexaacyl: 20 per cent to 40 per cent; It complies with the test) carried out using 10 mL for each
~ heptaacyl: less than 0.5 per cent. medium.
Triethylamine Congener distribution
4.2 to 5.8 per cent mlm, determined by a suitable method, The relative amount of tetraacyl, pentaacyl, hexaacyl and
for example gas chromatography (2.2.21!). heptaacyl congener groups are determined by a suitable
Water (2.5.12) method) for example reversed-phase HPLC analysis (2.2.29).
Maximum 6.7 per cent mtm, The relative amount of each congener group in the 3-0-
desacyl-4'-monophosphoryllipid A liquid bulk is:
Free fatty acids
- tetraacyl: 15 per cent to 35 per cent;
Maximum 2.6 per cent mlm, determined by a suitable
- pentaacyl: 35 per cent to 60 per cent;
method, for example reversed-phase HPLC analysis (2.2.29).
- hexaaeyl: 20 per cent to 40 per cent;
2..Keto-3-deoxyoctonate - heptaacyJ: less than 0.5 per cent.
Less than 0.5 per cent mlm, determined by a suitable
method. For example, a colorimetric method may be used ASSAY
where 2-keto-3-deoxyoclOnate is released by hydrolysis (0.2 The 3-Q-desacyl-4'-monophosphoryllipid A content is
N H,SO. at 100°C for 30 min), oxidised by periodic acid, determined by a suitable method) for example gas
and reacted with sodium arsenite to yield p-fonnylpyruvic chromatographic quantification (2.2.28) of trifluoroacetic
acid) which subsequently is coupled to thiobarbituric acid to anhydride derivatised fany acid methyl esters of the 3-D-
give a red coloured chromophore with absorption maximum desacyl-4'-monophosphoryllipid A futty acids dodecanoic
at 550 nm. The amount of 2-keto-3-deoxyoctonate is acid (CI2:0), tetradecanoic acid (CI4:0), 3-hydroxy
interpolated from a calibration curve. tetradecanoic acid (3-0H-CI4:0) and hexadecanoic acid
(CI6:0) obtained by hydrolysis of 3-0-desacyl-4'-
Identity monophosphoryl lipid A in an aqueous/methanol (50:50 VII')
The test for congener distribution also serves to identify the solution) containing 5 per cent of sodium hydroxide. To the
product. test sample) a reference sample and the dilutions of the
Microbial contamination calibration curve) pentadecanoic acid (CI5:0) is added as an
TAMC: acceptance criterion 10' CFU/IO mg (2.6.12). internal standard. The temperature gradient applied must
Pyrogens (2. 6.I!) allow the separation of the fatty acid methyl esters in about
The triethylamine salt of 3-D-desacyl-4'-monophosphoryl 40 min.
lipid A complies with the test for pyrogens. Inject into each The sum of the ratios between the area for each individual
rabbit per kilogram of body mass 3 mL of a solution farty acid methylester (CI2:0, CI4:0, 3-0H-CI4:0 and
containing 2.5 ~g of 3-0-desacyl-4'-monophosphoryllipid A. =
C 16:0) and the area of the internal standard (ratio area
3-0-DESACYL-4'-MONOPHOSPHORYL LIPID A C. I area CI5:0) is calculated. The 3-0-desacyl-4'-
LIQUIDBUU< monophosphoryl lipid A quantity corresponding to the sum
The triethylamine salt of 3-D-desacyl-4'-monophosphoryl ratio value on the calibration curve, established with the
lipid A is dispersed in a liquid suitable for the subsequent dilutions of the 3-D-desacyl-4'-monophosphoryllipid A
processing steps at a defined target concentration. If the salt standard) is reported.
is not soluble in water a microfluidisation step is necessary to The content of 3-0-desacyl-4'-monophosphoryllipid A is
prepare a stable aqueous suspension. not less than 80 per cent and not greater than 120 per cent
The liquid bulk is sterilised by filtration through a bacteria- of the estimated content.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
retentive filter.
Only a 3-0-desacyl-4'-monophosphoryllipid A liquid bulk
mat complies with the requirements given below under
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2022 Desferrioxamine Mesilate 1-733
Desferrioxamine Mesilate *** pH (2.2.3)

*** *** 4.0 to 6.0.
(Deferoxamine Mesllau, Ph. Bur. monograph 0896) *** Dilute I mL of freshly prepared solution S to 10 mL with
carbon dioxide-free waterR prepared from distilled water R.
o OH Related substances
HO"N~~~~~NH2 Liquid chromatography (2.2.29). Prepare the solutions
immediately before use and protect /rom light.
¢~ ?H 0
SOaH
I
CH,
A. Deferoxamine mesilate produced by fermentation.
Solvent mixture acctm1itrile R, waterR (6:94 VII').
HN~NyCH3 Test solution Dissolve 25 mg of the substance to be
examined in 10 mL of the solvent mixture and dilute to
o 25.0 mL with the solvent mixture.
C,o!f"N,O"S 657 138-14-7 100.0 mL with the solvent mixture.
Reference solution (b) Dissolve 5 mg of deferoxamine for
Action and use
system suitability CRS (containing impurity A) in mobile
Chelating agent (iron).
phase A and dilute to 5 mL with the solvent mixture.
Preparation
Column:
Desferrioxamine Injection
=
- size: I :::; 0.075 m, 0 4.6 rnm;
PIlE" _ - stationary phase: octadecylsilyl silica gelfor
chromarography R (3.5 ~m);
DEFINITION - temperature: 32 "C.
N'-[5-(4-[(5-Aminopentyl)(hydroxy)amino)-4-
Mobile phase:
oxobutanamido)pentylj-N'-hydroxy-N'-[5-(N-
- mobile phase A: 1.32 gIL solution of ammonium
hydroxyacetamido)pentyl]butanediamide methanesulfcnate.
phosphate R adjusted to pH 3.0 with phosphoric acidR;
Content - mobile phaseB: mixture of equal volumes of acetonitrile
98.0 per cent to J 02.0 per cent (anhydrous substance). for chromatography R and mobile phase A;
PRODUCTION
It is considered that alkyl methanesulfonate esters are Time Mobile phase A Mobile phase B
(min) (per cent VIJI) (per cent VA?
genotoxic and are potential impurities in deferoxarnine
0-4 88 12
mesilate. The manufacturing process should be developed
taking into consideration the principles of quality risk 4·20 88 ..... 80 12 ..... 20
20 - 35 80 --J 57.5 20 ..... 42.5
management) together with considerations of the quality of
starting materials, process capability and validation.
The general methods 2.5.37. Methyl, ethyland isopropyl Flow rate 1.5 mllmin.
methanesulfonate in methanendfonic acid, 2.5.38. Methyl, ethyl Detection Spectrophotometer at 220 nm.
and isopropyl methanesulfonate in active substances and Injection 20 ~L.
2.5.39. Methanesulfonyl chloride in methanesulfonic acid are
Identification of impwities Use the chromatogram obtained
available to assist manufacturers.
CHARACTERS impurity A.
Appearance Relativeretention With reference to deferoxamine (retention
White or almost white powder. time =about 18 min): impurity A =about 0.9.
Solubility System suitability Reference solution (b):
Freely soluble in water, slightly soluble in methanol, very - resolution: minimum 2.0 between the peaks due to
slightly soluble in ethanol (96 per cent). impurity A and deferoxamine.
IDENTIFICATION Cotcakuion .of percentage contents:
A. Infrared absorption spectrophotometry (2.2.24). - for each impurity, use the concentration of
Comparison deferoxamine mesilate CRS. deferoxamine mesilate in reference solution (a).
B. Dissolve 0.1 gin 5 mL of daUle hydrochlmU acidR. Limits:
Add 1 mL of bonum chloride solutron R2. The solution is - impurity A: maximum 2.0 per cent;
clear. In a porcelain crucible, mix. 0.1 g with 1 g of anhydrous - any otherimpun'ty: for each impurity, maximum
sodium carlxmate R, heat and ignite over a naked flame. Allow 0.8 per cent;
to cool. Dissolve the residue in 10 mL of water R, heating if - total: maximum 5.0 per cent;
necessary, and filter. The filtrate gives reaction (a) of sulfates - reporting threshold: 0.05 per cent.
(2.3.1). The thresholds indicated under Related substances
TESTS
pharmaautical use (2034) do not apply.
Solution S
Dissolve 2.5 g in carbon dioxide-free water R prepared from B. Deferoxamine mesilate produced by a synlhetic process.
distilled waterR and dilute to 25 mL with the same solvent. Solvent mixture Mobile phase B, mobile phase A
(20:80 VII').
than reference solution Y5 (2.2.2, MethodIf).
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1-734 Desferrioxamine Mesilate 2022
Test solution Dissolve 0.125 g of the substance to be - reporting threshold: 0.03 per cent.
examined in the solvent mixture and dilute to 10.0 mL with The thresholds indicated under Related substances
the solvent mixture. (Table 2034.-1) in the general monograph Subsumces for
Reference solution (a) Dilute 1.0 mL of the test solution to pharmaceutical use (2034) do not apply.
100.0 mL with the solvent mixture. Dilute 1.0 mL of this Chlorides (2.4.4)
solution to 10.0 mL with the solvent mixture. Maximum 330 ppm.
Reference solution (b) Dissolve 12.5 mg of deferoxamine for Dilute 2 rnL of solution S to 20 mL with water R.
peak identification CRS (containing impurities F, G, H, I
Sulfates (2.4. 13)
and J) in the solvent mixture and dilute to 1 mL with the
Maximum 400 ppm.
solvent mixture.
Column: Dilute 5 mL of solution S 10 20 mL with distined waterR.
- size: 1= 0.15 m, 0 = 4.6 mm; Water (2.5.12)
- stationary phase: end-capped rxtaduylsilyl silica gelfor Maximum 2.0 per cent, determined on 1.000 g.
chromOlcgraphy R (5 urn); Sulfated ash (2.4.14)
- temperature: 25°C. Maximum 0.1 per cent, determined on 1.Q g.
Mobile phase: Bacterial endotoxins (2.6. 14)
- mobile phase A: dissolve 0.605 g of sodium edetate R in Less than 0.025 Wlmg, if intended for use in the
900 mL of waterfor chromatography R, add 1.0 mL of manufacture of parenteral preparations without a further
phosphoric acidR and adjust to pH 6.0 with appropriate procedure for the removal of bacterial
concentrated ammonia R; dilute to 1000 mL with water endotoxins. Since deferoxamine mesilate has an inhibitory
for chromatography R; effect on the bacterial endotoxins test, a suitable procedure is
- mobile phaseB: dissolve 0.780 g of sodium edetate R in put in place to remove this inhibitory effect. The monocyte-
750 mL of waterfor chromatography R and add 1.0 mL activation test (2.6.30) has been found suitable to overcome
of phosphoric add R; add 250 mL of acetonitrile for this issue.
chromawgraphy R, mix thorougWy and adjust the
apparent pH to 6.0 with concentrated ammonia Rj ASSAY
Dissolve 0.500 g in 50 mL of waterR. Add 4 mL of dilute
Time MobUe phase A Mobile phase B sulfuric acid Rl, Titrate with 0.1 AtI feme ammonium sulfate.
(min) (per cent VIJI) (per cent Vm To accelerate the titration, add 4.5 mL quickly, stop for
0 80 20 1.5 min and continue to titrate uniformly at a rate of about
o. 21.6 80 ~ 20 20 -+ 80 0.2 mUmin. Determine the end-point potentiometrically
21.6 - 31.6 20 80 (2.2.21J) using a platinum indicator electrode and a suitable
reference electrode.
Flew rate 1.5 mUmin. 1 mL of 0.1 M ferne ammonium sulfate is equivalent to
65.68 mg ofC,oH52N.OIlS,
Injection 20~. STORAGE
Protected from light. If the substance is sterile, store in a
Identification of impun"ties Use the chromatogram supplied
with dejeroxamine for peak identification CRS and the
chromatogram obtained with reference solution (b) to LABELLING
identifythe peaks due to impurities F, G, H, I and J. The label states the origin of the substance:
Relative retention With reference to deferoxamine (retention - produced by fermentation;
time = about 12.9 min): impurity I = about 0.5; - produced by a syntheticprocess.
impurity F = about 0.96; impurity G = about 0.98; IMPURlTIES
impurity H = about 1.2; impurities J and K = about 1.37. Test A for related substances: A, B, C, J.
System suitability Reference solution (b): Test B for related substances: E, F, G, H, I, J, K.
- peok-to-ualiey ratio: minimum 3.0, where H p = height
Specified impurities A, F, G, H, I, J, K.
above the baseline of the peak due to impurity G and
H\J = height above the baseline of the fewest point of Otherdetectable impurities (the following substances would, if
the ClU'Ve separating this peak from the peak due 10 present at a sufficient level, be detected by one or other of the tests
deferoxamine. in the monograph. They are limited by the general acceptance
criterion for otherlumpecified impurities. It is therefore not
Calculation of peranUIge contents:
necessary l() identify these impun'lies for demonstration of
- correction factor: multiply the peak area of impurity I by
1.4;
phannaceutica/ use) B, C, E.
deferoxamine mesilate in reference solution (a).
Limits:
- impurities P, H: for each impurity, maximum
0.5 per cent;
- impurities OJ 1: for each impurity, maximum
0.3 per cent,
- sum of impurities J and K: maximum 0.5 per cent,
- unspecified impun'ues: for each impurity, maximum
0.20 per cent;
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2022 Desferrioxamine Mesilate 1-735
o OH
HO"'N~~~~~NH2
¢~ 9H 0
HN~NyCH3
A. N'-[5-(4-[(4-aminobuty1)(bydroxy)aminoj-4-
oxobutanamido)pentylj-NI-hydroxy-N'-[5-(N-
hydroxyacetamido)pentyljbutanediamide,
H. N' ,N"-bis(5-aminopentyl)-N',N",11,27-tetrahydroxy-
4,12,15,23,26,34-hexaoxo-5 ,11,16,22 ,27 ,33-
hexaazaheptatriacontane-I ,37 -diamide,
B. N-(5-aminopentyl)-N-hydroxy-N'-[5-(N-
hydroxyacetamido)penty1lbutanediamide,
K O
OH
/(N~~yCH3
o 0
I. N'-(5-aminopentyl)-N'-[5-(4-[(5-aminopentyl)(bYdroxy)
aminoj-4-oxobutanamido)pentylj- N'-
C. N-[5-(2,5-dioxopyrrolidin-I-yl)pentylj-N- hydroxybutanediamide,
hydroxyacetamide,
o OH
HO"'N~~~N~NH2
¢~ 9 H
HN~N. ~ .JlN~NJ...CH3
: 0
'( <;» -H I
o OH
E. methyl 4-[[5-[[4-[(5-aminopentyl)(bydroxy)amino]-4- J. N'-(5-aminopentyl)-N1,II ,22-trihydroxy-N"6-[5-(N-

oxobutanamidojpentyl] (bydroxy)aminoj-4-oxobutanoate, hydroxyacetamido)pentyIJ-4,12,15,23-tetraoxo-5,11,16,22-
tetraazahexacosane-I,26-diamide,
o OH
HO'N~~~~~NH2
¢~ H 0
HN~NyCH3
F. N'-(5-acetamidopentyl)-N'-[5-[4-[(5-
aminopentyl) (bydroxyl)aminoj-4-oxobutanamidojpentylj-
N'-hydroxybutanediamide,
G. unknown structure,
K. 22-(acetyloxy)-N1,N13_bis(5_aminopentyl)_N' ,N",11,33-
tetrahydroxy-4,12J 15,29,32,40-hexaoxo-
5,11,16,22,28,33,39-heptaazatriterracomane-1,43-diamide.
____________________ ""'11
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1-736 Desipramine Hydrochloride 2022
Reference solution (b) Dissolve 5 mg of imipramine

Desipramine Hydrochloride hydrochloride R (impurity A) in mobile phase A and dilute to
(Ph. Eur. monograph 0481) 100 mL with mobile phase A . .Mix 1 mL of the solution and
9 mL of the test solution, and dilute to 100 m1 with mobile
phase A.
Column:
, Hel - size: 1= 0.25 m, 12) = 4.6 nun;
- suuionatyphase: base-deactivated end-capped oaodecylsilyl
silica gelfor chromatography R (5 urn);
Mobile phase:
302.8 58-28-6 - mobile phaseA: methanol R, acetonitrile R, buffer solution
(11:14:75 VIVIV);
Action and use - mobile phaseB: methanol R, acetonitrile R, buffer solution
Monoamine reuptake inhibitor, tricyclic antidepressant. (28:34:38 VIVIV);
Preparation
Desipramine Tablets Tim. Mobile phase A Mobile phase B
(min) (per cent VIII) (per cent V/V,)
PhEIJ ~ _
0-2 85 15
DEFINITION 2 - 37 85 -+ 0 15 -+ 100
3-(10,II-Dihydro-5H-dibenzo[bJlazepin-5-yl)-N- 37 - 52 0 100
mel:hylpropan-I-amine hydrochloride.
Content FWw rate 1.4 mUmin.
99.0 per cent to 101.0 per cent (dried substance). Detection Spectrophotometer at 254 nm.
CHARACTERS Injection 40 ~L.
Appearance Identification of impurities Use the chromatogram obtained
White or almost white, crystalline powder. with reference solution (b) to identify the peak due to
impurity A.
Solublllty
Sparingly soluble in water and in anhydrous ethanol, Re/alive retention With reference to desipramine (retention
practically insoluble in heptane. =
time about 19 min): impurity A about 1.7. =
mp System suitability Reference solution (b):
About 214 "C. - resolution: minimum 25 between the peaksdue to
desipramine and impurity A.
IDENTIFICATION
Calculation of percentage conte1!ts:
- for each impurity, use the concentration of desipramine
Comparison desipramine hydrochloride CRS. hydrochloride in reference solution (a).
B. It gives reaction (a) of chlorides (2.3.1). Limits:
Solution S 0.10 per cent;
Dissolve 1.25 g in carbon dioxide-free water R, warming to not - total: maximum 0.5 per cent;
more than 30 QC if necessary, and dilute to 25 mL with the - reporting threshold. 0.05 per cent.
same solvent. Loss on drying (2.2.32)
Appearance of solution Maximum 0.5 per cent, determined on 1.000 g by drying in
SolutionS, examined immediately after preparation,' is not an oven at 105°C.
more intensely coloured than reference solution BY6 (2.2.2, Sulfated ash (2.4.14)
Method II). Maximum 0.1 per cent, determined on 1.0 g.
Acidity or alkaIlnity ASSAY
To 10 mL of solution S add 0.1 mL of methyl red sdution R Dissolve 0.250 g in a mixture of 5 mL of 0.01 M hydrochloric
and 0.3 mL of 0.01 M sodium hydroxide. The solution is acid and 50 mL of ethanol (96 per cenl) R. CarTY out a
yellow. Not more than 0.5 mL of o. 01 M hydrochlorit: acid is potentiometric titration (2.2.2U), using 0.1 M sodium
required to change the colour of the indicator to red. hydroxide. Read the volume added between the 2 points of
Related substances inflexion.
Liquid chromatography (2.2.29). I mL of 0.1 M sodium hydroxide is equivalent to 30.28 mg of
Buffer solution Dissolve 5.2 g of dipotassium hydrogen C u,H'3CIN,.
phosphate R in 1000 mL of waterfor chromalOgraphy R, add STORAGE
I mL of triethylamine R and adjust to pH 6.4 with phosphoric Protected from light.
acidR.
Test solution Dissolve 50.0 mg of the substance to be IMPURITIES
examined in mobile phase A and dilute to 100.0 mL with Otherdaeaabie impurities (the following subsumces would, if
mobile phase A. present at a sufficient level, be detected by one or other of the tests
Reference solution (a)
monograph Sub'Ulnces for pharmaceutical use (2034). It is
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2022 Desflurane 1-737
therefore not necessary to identify these impurities for of methylene chloride R (impurity E) to the solution) using an
demonstration of compliance. See also 5.10. Control of impurities airtight syringe) and dilute to 50.0 mL with the substance to
in substances for pharmaceuticaluse) A. be examined. Dilute 5.0 mL of this solution to 50.0 mL with
the substance to be examined. Store at a temperature below
10 "C.
to 50.0 mL with the substance to be examined. Store at a
temperature below 10°C.
to 25.0 mL with the substance to be examined. Store at a
A. 3-(IO,II-dihydro-5H-dibenzo[b,j]azepin-5-yl)-N,N- temperature below 10°C.
dimethylpropan-l-amine (imipramine). Column:
__________ ~ P1IEII - material: fused silica;
- size: 1= 105 m, 0 = 0.32 mm,
- stationary phase: trifluoropropylmelhylpolysiloxane R (film
thickness 1.5 urn}.
Desflurane Carrier gas helium for chromatography R.
(Ph. Bur. monograph 1666) Splitratio 1:25.
F H F
Temperature:
- column: 30°C;
J...~F
F 0' A and eoantiomer
- injection port: 150°C;
F F - detector: 200 "C.
Deieaion Flame ionisation.
C,H,F,O 168.0 57041-67-5 Injection 2.0 ~L.
PhEIl _
Run time 35 min.
DEFINITION Relativeretention With reference to desflurane (retention
(2RS) -2-(Difluoromethoxy)-I, I, 1,2-tetraffuoroethane. = =
time about 11.5 min): impurity C about 1.06;
CHARACTERS =
impurity D = about 1.09; impurity A about 1.14;
Appearance
= =
impurity G about 1.39; impurity E about 1.5;
Clear, colourless, mobile, heavy liquid.
=
impurity B = about 1.7; impurity F about 2.2;
=
Solubility
Practically insoluble in water, miscible with anhydrous - number of theoretical plates: minimum 20 000, calculated
ethanol. for the peak due to impurity Aj
Relative density - symmetry factor: maximwn 2.0 for the peak due to
1.47, determined at 15 "C. impurity B.
bp Limits:
About 22 "C. - impun''Y B: not more than the difference between the area
IDENTIFICATION of the corresponding peak in the chromatogram obtained
with reference solution (a) and the area of the
Preparation Examine the substance in the gaseous state. the test solution (0.2 per cent VIV);
Comparison Ph. Eur. reference spearum of desfiurane. - impurity A: not more than the difference between the area
TESTS of the corresponding peak in the chromatogram obtained
The substance to be examinedmust be cooled to a temperature with reference solution (a) and the area of the
be/ow 10°C and the tests must be carried out at a temperature corresponding peak in the chromatogram obtained with
below 20 "C. the test solution (0.1 per cent VIV);
- impuruies C, D) G: for each impurity, not more than the
AcIdity or alkallnlty difference between the area of the peak due to impurity A
To 20 mL add 20 mL of carbon dioxide-free warer R, shake
for 3 min and allow to stand. Collect the upper layer and and the area of the peak due to impurity A in the
add 0.2 mL of bromocresol pwple solution R. Not more than chromatogram obtained with the test solution
0.1 mL of 0.01 M sodium hydroxide or 0.6 mL of 0.01 M
(0.01 per cent VIII);
hydrochloric add is required to change the colour of the - impuruies E) H: for each impurity) not more than the
indicator. difference between the area of the corresponding peak in
Related substances the chromatogram obtained with reference solution (a)
Gas chromatography (2.2.28). and the area of the corresponding peak in the
Test sdution The substance to be examined. chromatogram obtained with the test solution
Reference solution (a) Introduce 25 mL of the substance to (0.01 per cent VIII);
be examined into a 50 mL flask fitted with a septum) and - impurity F: not more than the difference between the area
add 0.50 mL of desflurane impurity A CRS and 1.0 mL of of the corresponding peak in the chromatogram obtained
isoflurane CRS (impurity B). Add 50 ~L of acewne R with reference solution (a) and the area of the
(impurity H), 10 ~L of chloroform R (impurity F) and 50 ~L
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1-738 Deslanoside 2022

the test solution (0.002 per cent VIV);
0.5 times the difference between the area of the peak due c. dichlorofluoromethane,
to impurityA in the chromatogram obtained with
reference solution (b) and the area of the peak due to
impurity A in the chromatogram obtained with the test
solution (0.005 per cent VIII);
- sumof impurities other thanA, B, C, D, E, F, G and H: D. trichIorofluoromethane,
not more than the difference between the area of the peak
due to impurity A in me chromatogram obtained with
reference solution (b) and the areaof the peak due to
impurity A in Ute chromatogram obtained with the test
solution (0.01 pe' cent VIII); E. dichloromethane (methylene chloride),
- disregard limit: the difference between the area of the peak
due to impurityA in the chromatogram obtained with
referencesolution (c) and the area of the peak due to
impurity A in the chromatogram obtainedwith the test
solution (0.002 pe' cent VIII). F. trichloromethane (chloroform),
Fluorides
Maximwn 10 ppm.
Potenrlomerry (2.2.36, Method I}.
Testsolution To 10.0 mL in a separating funnel, add 10 mL
of a mixture of 30.0 mL of dilute ammonia R2 and 70.0 mL G. 1,1,2-trichloro-I,2,2-triftuoroethane,
of distilled water R. Shake for I min and collect the upper
layer. Repeat this extraction procedure twice, collectingme
upperlayereach time. Adjust the combined upperlayers 10
pH 5.2 with dilute hydrochlori< acid R. Add 5.0 mL offiucride
'0
standard solution (/ ppm F) R and dilute 50.0 mL with
H. propan-2-one (acetone).
distilled water R. To 20.0 mL of this solution add 20.0 mL of
'0
total-ionic-strength-adjustment-buffer R and dilute 50.0 mL ______ ~ PhE"
with distilled water R.

Reference soltaions To each of 1.0 ml., 2.0 mL, 3.0 mL,
4.0 mL and 5.0 mL ei ftuotide standard solution (/0 ppm F) R
add 20.0 mL of total-ionic-strength-odjustment buffer R and Deslanoside
dilute '050.0 mL with distilled water R.
Indiauor electrode Fluorideselective.
Reference electrode Silver-silver chloride. o
Carry out the measurements on 20 mL of each solution.
Calculate the concentration of fluorides using the calibration
curve, taking into account the additionof fluoride to the test
solution.
Non-volatile matter
Maximwn 100 mgIL.
Evaporate 20.0 mL to dryness with the aid of a stream of
nitrogen R. The residue weighs not more than 2.0 mg.
STORAGE
In a glass bottle fitted with a polyethylene-lined cap. Before
opening the bottle, cool the contents to below 10°C.
IMPURITIES
Specified impuruies A, B) C, D, E) F, G) H.
943 17598-65-1
A. 1,1/-oxybis[(IS)-I,2,2,2-tetrafluoroethane], Action and use

NaIK-ATPase inhibitor; cardiac glycoside.
F H CI
FAo~F
PhE" ~ _
and enanlJomer
F F DEFINITION
Deslanoside contains not less than 95.0 per cent and not
B. (2RS) -z-chloro-2-(difluorornethoxy) -I, I, I-trifluoroethane more than the equivalent of 105.0 per cent of 3~-[(O-~-D
(isofluorane), glucopyranosyl-(1->4)-o.2,6-dideoxy-~-D-rib<>-hexopyranosyl-
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2022 Desloratadine 1-739
(I ~ 4)-o-2,6-dideoXY-~-D-rib<>-hexopyranosyl-(1 ~4)-2,6 Reference solution (c) Dilute 1 mL of reference solution (a)

dideoxy-B,D-rib<>-hexopyranosyl) oxy] -12~, 14-dihydroxy- to 10 mL with a mixture of equal volumes of chlorojonn R
5~,14~-card-20(22)-enolide, calculated with reference to the and methanol R.
dried substance. Apply separately to the plate as 10 mm bands 5 ~L of each
CHARACTERS solution. Develop immediately over a path of IS em using a
A white or almost white, crystalline or finely crystalline mixture of 3 volumes of water R~ 36 volumes of methanol R
powder, hygroscopic, practicaUy insoluble in water, very and 130 volumes of methylene chloride R. Dry the plate in a
slightly soluble in alcohol. In an atmosphere of low relative current of warm air, spray with a mixture of 5 volumes of
humidity, it loseswater. sulfuric acid Rand 95 volumes of alcohol R and heat at
140 °C for 15 min. Examine in daylight. In the
IDENTIFICATION
chromatogram obtained with test solution (a), any zone,
apart from the principal zone, is not more intense than the
Second identification: B, CJ D. zone in the chromatogram obtained with reference
A. Examine by infrared absorption spectrophotometry solution (b) (2.5 per cent) and at most two such zones are
(2.2.24), comparing with the spectrum obtained with more intense than the zone in the chromatogram obtained
deslanoside CRS. When comparing the spectra, special with reference solution (c) (1.0 per cent).
attention is given to the absence of a distinct absorption Loss on drying (2.2.32)
maximum at about 1260 cm-1 and to the intensity of the Not more man 5.0 per cent, determined on 0.500 g by
absorption maximum at about 1740 em-I. Examine the drying in vacuo at 105°C.
substances in discs prepared by dissolving 1 mg of the
substance to he examined or I mg of the reference substance
in 0.3 mL of methanol R and triturating with about 0.4 g of Not more than 0.1 per cent, determined on the residue
dry, finely powdered potassium bromide R until the mixture is obtained in the test for loss on drying.
uniform and completely dry. ASSAY
B. Examine the chromatograms obtained in the test for Dissolve 50.0 mg in alcohol R and dilute to 50.0 mL with the
related substances. The principal zone in the chromatogram same solvent. Dilute 5.0 mL of this solution to 100.0 mL
obtained with test solution (b) is similar in position, colour with alcohol R. Prepare a reference solution in the same
and size to the principal zone in the chromatogram obtained manner, using 50.0 mg of deslonoside CRS (undried).
with reference solution (a). To 5.0 mL of each solution add 3.0 mL of alkaline sodium
C. Suspend about 0.5 mg in 0.2 mL of alcohol picrate solution R and allow to stand protected from bright
(60 per <enr VIV) R. Add 0.1 mL of dinitrobenzoic acid light in a water-bath at 20 ± 1 °C for 40 min. Measure the
solution Rand 0.1 mL of dilute sodium hydroxide solution R. absorbance (2.2.25) of each solution at the maximum at
A violet colour develops. 484 om, using as the compensation liquid a mixture of
3.0 mL of a/haline sodium picrate solulion R and 5.0 mL of
D. Dissolve ahout 5 mg in 5 mL of glacial acetic acid R and
akohol R prepared at the same time.
add 0.05 mL offerric chloride solution Rl. Cautiously add
2 mL of sulfuric acid R~ avoiding mixing the two liquids. Calculate the content of C47H74019 from the absorbances
Allow to stand; a brown but not reddish ring develops at the measured and the concentrations of the solutions.
interface and a greenish-yellow, then bluish-green colour STORAGE
diffuses from it to the upper layer. Store in an airtight, glass container, protected from light, at a
TESTS temperature below 10°C.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
Solution S
Dissolve 0.20 g in a mixture of equal volumes of chlor%nn R
and methanol R and dilute to 10 rnL with the same mixture
of solvents.
Appearance of solution Desloratadine
Solution S is clear (2.2.1) and colourless (2.2.2, Method 11).
Speclllc optical rotation (2.2.7)
Dissolve 0.200 g in anhydrous pyridine R and dilute to
10.0 mL with the same solvent. The specific optical rotation
is + 6.5 to + 8.5~ calculated with reference to the dried
substance.
Related substances
310.8 100643-71-8
Test solution (a) Use solution S.
Test solution (b) Dilute I mL of test solution (a) to 10 mL Action and use
with a mixture of equal volumes of chlorototm Rand Histamine Hl, receptor antagonist; antihistamine.
methanol R. PhE" ~ _
Reference solution (a) Dissolve 20 mg of deslanoside CRS in a
mixture of equal volumes of chlorojonn Rand methanol Rand DEFINITION
dilute to to mL with the same mixture of solvents. 8-Chloro-ll-(piperidin-4-ylidene)-6,II-dihydro-5H-benzo
Reference solution (b) Dilute 2.5 mL of reference solution (a) [5,6] cyclohepta[ 1,2-b]pyridine.
to 10 mL with a mixture of equal volumes of chlorofonn R Content
and methanol R. 98.0 per cent to 102.0 per cent (anhydrous substance).
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1-740 Desloratadine 2022
CHARACTERS - for each impurity, use the concentration of desloratadine

Appearance in reference solution (b)"
White or almost white powder. Limits:
Solubility - impun"ty B: maximum 0.3 per cent;
Very slightly soluble or practically insoluble in water, freely - impun'cy A: maximum 0.2 per cent;
soluble in ethanol (96 per cent), slightly soluble or very - unspecified impurities: for each impurity, maximum
slightly soluble in heptane. 0.10 per cent;
- IOtal: maximum 0.4 per cent;
IDENTIFICATION
Water (2.5.12)
Comparison desloratadine CRS.
If me spectra obtained in the solid state show differences, Maximum 0.2 per cent, determined on 0.5 g.
substance separately in methylisobutyl ketone R, evaporate lO ASSAY
dryness and record new spectra using the residues. Liquid chromatography (2.2.29) as described in the test for
related substances with the foUowing modifications.
TESTS
Injection Test solution and reference solution (a),
Related substances
Liquid chromatography (2.2.29). System suitability Reference solution (a):
- symmetry factor: 0.5 to 1.5 for the peak due to
desloraradine.
examined in the mobile phase and dilute (0 25.0 mL with
the mobile phase. Dilute 5.0 mL of the solution to 50.0 mL Calculate the percentage content of Cl9H19CIN2 takinginto
with the mobile phase. account the assigned content of desloratadine CRS"
Reference solution (a) Dissolve 20.0 mg of desloratadine CRS IMPURITIES
in the mobile phase and dilute to 25.0 mL with the mobile Specified impurities A, B.
phase. Dilute 5.0 mL of the solution to 50.0 mL with the Other detectable impun'Ues (the following substances uouid, if
mobile phase. present at a sufficient level., be detected by one or other of the tests
Reference solution (b) Dilute 1.0 mL of the test solution to in the monograph. Theyare limited by thegeneral acceptance
100.0 mL with the mobile phase. Dilute 1.0 mL of this criterion for other/Unspecified impurities and/or by thegeneral
solution to 10.0 mL with the mobile phase. monograph Subsumces for pharmaceutical use (2014). It is
Reference solution (c) Dissolve 4 mg of desloratadine for system therefore not necessary to identify these impumies for
suitab/7ity CRS (containing impurities A and B) in the mobile demonstration 'If compliance. See also 5.10. Control of impurities
phase and dilute to 5.0 mL with the mobile phase. Dilute it, substances for pharmaceutical use) C.
C't6F.OH
1.0 mL of the solution to 10.0 mL with the mobile phase.
Column:
- size: I = 0.25 m, 0 = 4.6 mID;
- stationary phase: end-capped ocUldecylsiiyl silica gelfor and enantlomer
chromatography R (4 pm), f' N
- temperature: 35 "C. ee-,
Mobile phase Dissolve 0.865 g of sodium dadecy/ sulfate R in
water R, add 0.5 rnL of tnJluoroacetic add R and dilute to A. (1IRS)-8-<:Woro-l l-ftuoro-ll-(piperidin-4-yl)-6,1 1-
1000 mL with water R; mix 57 volumes of this solution and dihydro-5H-benzo[5,6]cyclohep,a[I,2-b]pyridine,
43 volumes of acewnitrile R.
"~~
Detection Spectrophotometerat 280 run.
lnjution 100 J..IL of the test solution and reference •• erenecmer
Run time 2.5 times the retention rime of desloratadine.
Identification of impun"ues Use the chromatogram supplied
with desloratadine for system suitability CRS and the B. (I IRS)-8-<:hloro-ll-(1,2,3,6-tetrahydsopyridin-4-yI)-6,1 1-
chromatogram obtained with reference solution (c) to identify dihydro-5 H-benzo [5,6]cyclohepta[I ,2-b[pyridine,
the peaks due [0 impurities A and B.
Relative retention With reference to desloratadine (retention
System suiUlbility Reference solution (c):
impurity Band desloratadine.
Calculation of perCentage contents:
- correction factors: multiply the peak areas of the following C. ethyl 4-(8-chloro-5,6-dihydro-lIH-benzo[5,6]
impurities by the correspondingcorrection factor: cyclohepta [1,2-b]pyridin-II-ylidene)piperidine-l-
impuriry A = 1.6; impurity B = 1.6; carboxylate (loraradine).
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2022 Desmopressin 1-741
Desmopressin *** Test solution Dissolve 1.0 mg of the substance to be

*** *** examined in 2.0 mL of water R.
(Ph. Bur. monograph 0712) *** Resolution soluuon Dissolve the contents of a vial of
oxytocinldesmopressin validation mixture CRS in 500 J.lL of
~
water R.
Ty' - Phe - Gle -Me - ct - P,o - 0-"""- Gly- NH, Column:
- size: 1= 0.12 m, 0 = 4.0 nun;
o
- stationary phase: octade<ylsilyl silica gelfor chromatography R
1069 16679-58-6 (5 urn).
Mobile phase:
Action and use - mobile phase A: 0.067 M phosphate buffersolution pH 7.0 R;
Vasopressin analogue; treatment of diabetes insipidus; filter and degas;
nocturnal enuresis; haemophilia; von Willebrand's disease. - mobile phaseB: acetonitrile for chromatography R, mobile
Preparations phase A (50:50 VII'); filter and degas.
Desmopressin Injection
Desmopressin Intranasal Solution Time Mobile phase A Mobile phase B
(min) (per cent VIJI) (per cent VM
Desmopressin Nasal Spray
0-4 7' 24
Desmopressin Oral LyophiJisate "·18 76 --> 58 24 --> 42
Desmopressin Tablets 18 - 35 58 --> 48 42 --> 52
35 - 40 48 --> 76 52 --> 24
PhE" _
40 - 50 7. 24
DEFINITION
(3-Sulfanylpropanoyl)-L-tyrosyl-L-phenylaJanyl-L-glutaminyl- Flow rate 1.5 mUmin.
L-asparaginyl-L-cysteinyl-L-prolyl-D-arginylglycinamide cyclic Detection Spectrophotometer at 220 run.
(I ~6)-disulfide.
Injection 50 ~L.
Synthetic cyclic nonapeptide, available as an acetate.
Retention time Desmopressin = about 16 min;
Content oxytocin = about 17 min.
95.0 per cent to 105.0 per cent (anhydrous and acetic acid-
System suitability Resolution solution:
free substance).
CHARACTERS desmopressin and oxytoxin.
Appearance Limits:
White or almost white, fluffy powder. - unspecified impurities: for each impurity, maximum
Solubility 0.5 per cent,
Soluble in water, in ethanol (96 per cent) and in glacial - total: maximum 1.5 per cent;
acetic acid. - disregard limit: 0.05 per cent.
IDENTIFICATION Acedc acid (2.5.34)
A. Examinethe chromatograms obtained in the assay. 3.0 per cent to 8.0 per cent
Results The retention time and size of the principal peak in Test solution Dissolve 20.0 mg of the substance to be
the chromatogram obtainedwith the test solutionare examined in a mixture of 5 volumes of mobile phaseBand
approximately the same as those of the principal peak in the 95 volumes of mobile phase A and dilute to 10.0 mL with
chromatogram obtained with the reference solution. the same mixture of mobile phases.
B. Amino acid analysis (2.2.56). For hydrolysis use Method I Water (2.5.32)
and for analysis use Method 1. Maximum 6.0 per cent, determined on 20.0 mg.
Express the content of each amino acid in moles. Calculate Bacterial endotoxlns (2.6. I4)
the relative proportions of the amino acids, taking 1/6 of the Less than 500 IU/mg) if intendedfor use in the manufacture
sum of the number of moles of aspartic acid, glutamic acid) of parenteral preparations without a further appropriate
proline) glycine, arginine and phenylalanine as equal to 1. procedure for the removal of bacterial endotoxins.
The values fall within the following limits: aspartic acid: ASSAY
0.90 to 1.10; glutamic acid: 0.90 to 1.10; proline: 0.90 to Liquid chromatography (2.2.29) as described in the test for
1.10; glycine: 0.90 to 1.10; arginine: 0.90 to 1.10; related substances with the following modifications.
phenylalanine: 0.90 to 1.10; tyrosine: 0.10 to 1.05; half-
cystine: 0.30 to 1.05. Lysine) isoleucine and leucine are Reference solution Dissolve the contents of a vial of
desmopressin CRS in water R to obtain a concentration of
absent; not more than traces of otheramino acids are
0.5 mg/mL
presenr.
Mobile phase Mobile phase B, mobile phase A (40:60 VIV).
TESTS
Specific optical rotadon (2.2.7)
-12 to -82 (anhydrous and acetic acid-free substance). Retention time Desmopressin = about 5 min.
Dissolve 10.0 mg in a 1 per cent VIV solutionof glacial acetic Calculate the content of desrnopressin (C4JI6~14012S2)
acidR and dilute to 5.0 mL with the same acid. from the declared content of C46H6..N14012S2 in
desmopressin CRS.
Related substances
Liquid chromatography (2.2.29): use the oormalisation
procedure.
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1-742 Desogestrel 2022
s
~ GIy-~
STORAGE
I CH,
Tyr,phe'Gln' Asn- cys -Pro - 0-..... -
temperature of 2 °C to 8 "C. If the substance is sterile, store o CH3
in a sterile, airtight, tamper-evident container.
LABELLING G. N 1.9,NI.9-dimethyldesmopressin.
The label states: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
- the mass of peptide per container;
IMPURITIES Desogestrel
Owerdetectable impurities (the following mbstances would, if
present at a sufficient level, be detected by one or other of the WLS (ph. Eur. monograph 1717)
monograph Substances for pharmaceutical use (2034). I. is
H' C r
CffiP
H2C . OH
therefore not neassmy to idenufy these ;mpun"ties for H H
in substances for pharmaceutical use) A} B, C, D, E, F, G. ~ ~
d'
~
o
Tyr' Phe·GIn· Asp .l.p," -o·..... -Gly-NH,
Action and use
310.5 54024-22·5
Progestogen.
A. [5-L-aspartic acid]desmopressin, Preparation
Desogestrel Tablets
o
~Tyr'Phe'Glu'Asn.cl,p,"-o-"".-GIy-NH' PhE"
DEFINITION
_
Lj-Bthyl-Ll-methylldene-l S,19-dinor-17«-pregn-q-en-gn-yn-
B. [4-L-glutamic acidjdesmopressin, 17-01.
Content
~
o
Tyr' Ph.·Gln· Asn.l.pro - o-....g -GIy- OH CHARACTERS
Appearance
C. [9-glycine]desmopressin, SolublIlty
Practically insoluble in water, very soluble in methanol, freely
soluble in anhydrous ethanol and in methylene chloride.
o
~ Tyr 'Phe'G1n' Asn.l.p," -l·..... -Gly -NH,
IDENTIFICATION
Comparison desogestrel CRS.
D. [8-L-arginine]desmopressin, B. Specific optical rotation (see Tests).
TESTS
25.0 mL with the same solvent,
Related substances
E. N"'-[(acetylamino)methyljdesmopressin, Test solution Dissolve 20.0 mg of the substance to be
examined in 25 mL of aceronitrile R1 and dilute to 50.0 mL
with water R.
Reference solution (aJ Dissolve 4 mg of desogestrel for system
suitability CRS (containing impurities A, BJ C and D) in
5 mL of acetonitrile R1 and dilute to 10.0 mL with water R.
100.0 mL with a mixture of equal volumes of acetonitrile RJ
and water R.
F. N"'-[(acetylamino)methyljdesmopressin, Reference solution (c) Dilute 1.0 mL of reference solution (b)
to 10.0 mL with a mixture of equal volumes of acetonitrile RJ
and water R.
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2022 Desogestrel 1-743
Reference solution (dJ Dissolve 20.0 mg of desogestrel CRS in in me monograph. They are limited by the general acceptance
25 mL of acetonitrile Rl and dilute to 50.0 mL with waterR. oitetion for other/unspecified impurities and/or by the general
- size: 1;;:; 0.25 m, 0 = 4.6 mm, therefore not necessary to identify these impurities for
- stauimary phase: stericaUy protected ocladecylsi/yl silica gel demonstration of compl£ance. See also 5.10. Control oj impurities
fur chromatography R (5 urn), in substances for pharmaceutical use) E.
CH
Mobilephase waterR, acetonitrile R I (27:73 VIV). fl
~
H'C
Flow rate 1.0 mIJrnin. H2C ' OH
Detection Spectrophotometer at 205 om. H H
Injection 15 J1L of the test solution and reference ~ ~
solutions (a), (b) and (c). "" H
,
Run time 2.5 times the retention time of desogestrel,
with desogestrel fur system suitability CRS and the A. l3-ethyl-ll-methylidene-18, Ic-dinor-S«, l7~-pregn-3-en-
chromatogram obtained with reference solution (a) to 20-yn-17-o1 (desogestrel tI'-isomer),
identify the peaks due to impurities A, B, C and D.
CH
Relative retention With reference to desogesrrel (retention /fI
= =
time about 22 min): Impurity E about 0.2; 'C CH" OH
dHP
impurity D = about 0.25; impurity B = about 0.7; H H
= =
impurity A about 0.95; impurity C about 1.05. : ,
System suitability Reference solution (a): Ii Ii
..?
- peak-UJ-iJalley ratio: minimum 2.0) where Hp ;:: height
H v ;:: height above the baseline of the lowest point of the B. II-methylidene-19-nor-17~-pregn-4-en-20-yn-17 -01,

desogestrel. "'C O
@
H,C
Limits:
H H
- correction factors: for the calculation of content) multiply
the peak area of the following impurities by the ~ ~
corresponding correction factor: impurity A ;:: 1.8, ..?
impurity D = I. 5;
- impun"ties A) BJ C: for each impurity, not more than twice C. l3-ethyl-ll-methylidenegon-4-en-17-one,
r
obtained with reference solution (c) (0"2 per cent);
~
- impurity D: not more than the area of the principal peak H'C
in the chromatogram obtained with reference solution (c) H,C • OH
(0.1 per cent); H H
- unspecified impurities: for each impurity, not more than the ,, ,,
H H
o ..?
peak in the chromatogram obtained with reference D. l3-ethyl-17 -hydroxy-ll-methylidene-18,19-dinor-17~
solution (b) (0.5 per cent); pregn-4-en-2 O-yn-3-one,
it
~
(0.05 per cent). H'C
H2C • OH
Loss on drying (2.2.32) H H
vtU:uo at a pressure not exceeding 2 kl'a. ~ ~
H·· .#
Sulfated ash (2.4./4) HO
ASSAY E. l3-ethyl-ll-rnethylidene-18,19-dinor-17~-pregn-4-en-20
Liquid chromatography (2.2.29) as described in the test for yne-3~,17-diol.

related substances with the following modification. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE"
.Injection Test solution and reference solution (d).

Calculate the percentage content of C 22H30 0 from the areas
of the peaks and the declared content of desogestrel CRS.
IMPURITIES
Specified impurities A, B, C, D.
Other detectable impurities (the foUowing subst.anus would) if
present at a sufficient leoel, be detected by oneor other of the tests
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1-744 Dexamethasone 2022
C. Add about 2 mg to 2 mL of sulfuric acid R and shake to

Dexamethasone dissolve. Within 5 min, a faint reddish-brown colour
(Ph. Eur. monograph 0388) develops. Add this solution to 10 mL of water Rand mix; the
colour is discharged.
TESTS
Dissolve 0.250 g in anhydrous ethanolR and dilute to
o Related substances
Liquid chromatography (2.2.29). Cany out the test protected
392.5 50-02-2 from light.
Action and use Testsolution Dissolve 25.0 mg of the substance to be
Glucocorticoid. examined in 1.5 mL of acetonitrile R and add 5 mL of mobile
phase A. Sonicate until dissolution is complete and dilute to
Preparations 10.0 mL with mobile phase A.
Dexamethasone Eye Drops, Suspension
Reference solution (a) Dissolve 5 mg of dexamethasone for
Dexamethasone and Neomycin Ear Spray system suirability GRS (containing impurities B, F and G) in
Dexamethasone Tablets 0.5 mL of acetonitrile R and add 1 mL of mobile phase A.
Tobramycin and Dexamethasone Eye Drops, Suspension Sonicate until dissolution is complete and dilute to 2 mL
PhEIr ~ ~ _
DEFINITION 100.0 mL with mobile phase A. Dilute 1.0 mL of this
9-Fluoro-11 p,17,21-trihydroxy-16.-methylpregna-I,4-diene- solution to 10.0 mL with mobile phase A.
3,20-dione. Reference solution (c) Dissolve 5 mg of dexamethasone for
Content peak identification CRS (containing impurities J and K) in
97.0 per cent to J 03.0 per cent (dried substance). 0.5 mL of acetonitrile R and add 1 mL of mobile phase A.
Sonicate until dissolution is complete and dilute to 2 mL mL
CHARACTERS
Appearance .
White or almost white, crystalline powder. Column:
SoIublllty
- size: 1= 0.15 m, 0 4.6 mm;=
- slationary phase: end-copped IXtad«y/silyl silica gelfor
Practically insoluble in water, sparingly soluble in anhydrous chromawgraphy R (5 ~m);
ethanol, slightly soluble in methylene chloride. - temperature: 45 "C.
IDENTIFICATION Mobile phase:
Firstidentification: A, B. - mobile phase A: mix 250 mL of acetonitrile R and 700 mL
Second identification: B, C. of waterfor chromatography R and allow to equilibrate;
A. Infrared absorption spectrophotometry (2.2.24). dilute to 1000 mL with waterfor chromawgraphy R and
mix;
Comparison dexamethasone CRS.
- mob,1e phase B: acetonitrile R;
Test solution Dissolve 10 mg of the substance to be Time Mobile phase A Mobile phase B
examined in the mobile phase and dilute to 10.0 mL with (min) (per cent VIJI) (per cent VIJI)
the mobile phase. 0- 15 100 0
Reference solution Dissolve 10 mg of dexamethasone CRS in 15 - 40 100 -+ 0 o -+ too
phase. Flow Tale 1.2 mllrnin.
Plate TLC silica gel F". plateR. Detection Spectrophotometer at 254 om.
Mobile phase methanol R, meehylene chloride R (10:90 VIII). Injection 20 ~L; inject mobile phase A as a blank.
Application 5 ~L. Identification of impurities Use the chromatogram supplied
Development Over 3/4 of the plate. with dexamethasone for system sut'labili!)! CRS and'the
Drying In air. chromatogram obtained with reference solution (a) to
identify the peaks due to impurities B, F and G; use the
Detecdon Spray with a solution prepared as follows: dissolve
chromatogram supplied with dexamethasone for peak
0.25 g of 2,4-dihydroxybenzaldehyde R in glacial acetic acid R,
reference solution (c) to identify the peaks due to impurities J
12.5 mL of sulfuric acid Rand 37.5 mL of glacial acetic
andK.
add Rj heat at 90 °C for 35 min or until the spots appear,
allow to cool and examine in daylight and in ultraviolet light Relative retention With reference to dexamethasone
at 365 om. = =
(retention time about 15 min): impurity J about 0.90;
impurity B = about 0.94; impurity K = about 1.3;
impurity F = about 1.5; impurity G = about 1.7.
reference solution.
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2022 Dexamethasone 1-745
System suitability Reference solution (3):

o
- peak-to-valley ratio: minimum 2.0, where Hp = height
H" = height above the baseline of the lowest point of the
dexamethasone. o
Limits:
B. 9-lIuoro-II~, 17 .21-trihydroxy-16~-methylpregna-I,4
- impwiry G: not more than 3 times the area of the
diene-3,20-dione (betamethasone),
reference solution (b) (0.3 per cent); o
- impunhe.s H, P, J, K: for each impurity, not more than
(0.15 per cent);
- unspecified impurities: for each impurity, not more than the o
with reference solution (b) (0.10 per cent); C. c-fiuoro-t lp,17,21-trihydroxy-16~-methylpregn-4-ene
- total: not more than 5 times the area of me
principal peak 3,20-dione,
(0.5 per cent); o
(0.05 per cent).
an oven at 105°C.
D. 9,llll-epoxy-17,21-dihydroxy-16~-methyl-9Il-pregna-I,4
ASSAY diene-3,2o-dione,
Dissolve 0.100 g in ethanol (96 per cent) R and dilute to
solution to 100.0 mL with ethanol (96 per <en\! R. Measure
the absorbance (2.2.25) 'at the absorption maximum at
238.5 om.
Calculate the content of C22H29FOs taking the specific
absorbance to be 394. o
STORAGE E. 17,21-dihydroxy-l fu-methylpregna-I,4,9(l1)-niene-3,2Q-
Protected from light. dione,
IMPURITIES
Specified impurilies B, F, G, J, K.
Other deteaable impun·ties (the foIWwing substances would, if
present at a sufficient Ieuel, be detected by oneor other0/ the tests
craenon for other/unspecified impun·ties and/orby the general o
there/ore not necessary UJ identify these impurities for F. 9-lIuoro-11 ~,21-dihydroxy-16~-methylpregna-I,4-diene
demonstration of compliance. See also 5.10. Control of impun·ties 3,20-dione,
in substances for pharmaceutical use) A.J C.J D.J E, H.J 1.
o
o
G. 9-lIuoro-ll ~,17-dihydroxy-I6<x-methyl-3,20-dioxopregna
A. 14-lIuoro-ll~, 17 .21-trihydroxy-16~-methylpregna-I,4 1,4-dien-21-yl acetate (dexamethasone acetate),
diene-3,20-dione,
o
H. 17-hydroxy-I6<x-methyl-3,2Q-dioxopregna-I,4,9(II)-trien-
21-yl acetate,
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1-746 Dexamethasone Acetate 2022

Comparison dexamethasone acetate CRS.
substance separately in methylene chloride R, evaporate to
o dryness and record new spectra using the residues.
1. 9,llct-epoxy-17,21-dihydroxy-16ct-methylpregna-I,4- B. Thin-layer chromatography (2.2.27).
diene-3,20-dioneJ Testsolution Dissolve 10 mg of the substance to be
the mobile phase.
Reference solution Dissolve 10 mg of dexamethasone
acetate CRS in the mobile phase and dilute to 10.0 mL with
the mobile phase.
Plate TLC silica gel Fm plate R.
o
Mobile phase methanol R, methylme chloride R (10:90 VIV).
J. 17,21-dihydroxy-16ct-methylpregna-1 ,4-diene-3, 11,20- Applica'wn 5 ~L.
trione, Development Over 3/4 of the plate.
Drying In air.
Detection Spraywith a solutionprepared as follows: dissolve
0.25 g of 2,4-dihydroxybmzalde!ryde R in glacial acetic acid R,
12.5 mL of solfUlie acid Rand 37.5 mL of glacial acetic
o acid R; heat at 90°C for 35 min or until the spots appear,
allow to cool and examine in daylight and in ultraviolet light
K. 17,21-<lihydroxy-16ct-methylpregna-I,4,7,9(11)-terraene- at 365 nm.
3,20-dione. Results The principal spot in the chromatogram obtained
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PhE<I with the test solution is similar in position, colourand size to
reference solution.
c. Add about 2 mg to 2 mL of sulfuric acid R and shake to
Dexamethasone Acetate dissolve. Within 5 min, a faint reddish-brown colour
develops. Add this solution to 10 mL of water R and mix.
(Ph. Bur. monograph 0548) The colour is discharged and a clear solution remains.
D. Examine the chromatograms obtained in the assay.
Results The principal peakin the chromatogram obtained
with test solution (b) is similar in retention time and size to
the principal peak in the chromatogram obtainedwith
reference solution (e).
o TESTS
J177-87-3
434.5
Action and use 25.0 mL with the same solvent.
Glucocorticoid. Related substances
PhEu ~ __ Liquid chromatography (2.2.29). Cany ou' 'he test protected
from light.
DEFINITION Test solution (a) Dissolve 25.0 mg of the substance to be
9-Fluoro-11 p, 17-dihydroxy-16ct-methyl-3,20-dioxopregna- examined in about 4 mL of acetonitrile R and dilute to
1,4-dien-21-yl acetate. 10.0 mL with water R.
Content Tests%wn (b) Dilute 1.0 mL of test solution (a) to
97.5 per cenr to 102.0 per cenr (dried substance). 5.0 mL with the mobile phase.
CHARACTERS Reference solution (a) Dissolve 2 mg of dexamethasone CRS
Appearance (impurity A) and 2 mg of betamethasone acetate CRS
White or almost white, crystalline powder. (impurity D) in the mobilephase, using sonication for about
10 min, and dilute to 100 mL with the mobile phase.
Solubility
A1ix 1 mL of this solutionand 6 mL of test solution (a) and
dilute to 10 mL with the mobile phase.
(96 per cent), slighdy soluble in methylene chloride.
Reference sohuion (b) Dilute 1.0 mL of test solution (a) to
Ir shows polymorphism (5.9).
IDENTIFICATION solution to 10.0 mL with the mobile phase.
Fi,,' identification: A, D.
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2022 Dexamethasone Acetate 1-747
Reference solution (c) Dissolve the contents of a vial of J'tlobile phase acetonitrile R, water fQY' chromatography R
dexamethasone acetate impun"cy E CRS in 1 mL of the mobile (45:55 VIV).
phase. Injection Test solution (b) and reference solution (e).
Reference solution (d) Dissolve 5 mg of dexamethasone acetale Run time 1.5 times the retention time of dexamethasone
for peak identification CRS (containing impurity 1) in about acetate,
0.8 mL of acetonitrile R and dilute to 2 mL with water R. Retention time Dexamethasone acetate = about 13 min.
Reference solution (e) Dissolve 25.0 mg of dexamethasone Calculate the percentage content of C24H3lF06 taking into
acetate CRS in about 4 mL of acetoninile R and dilute to account the assigned content of dexamethasone acetate CRS.
10.0 mL with water R. Dilute 1.0 mL of the solution to
5.0 mL with the mobile phase. STORAGE
Column: Protected from tight.
- size: 1== 0.25 ID, 0 = 4.6 mm; IMPURITffiS
- stationary phase: end-copped extade<y/silyl silica gelfor Specified impurities A.. D, E, 1.
chromatography R (5 pm). Otherdetectable impuruies (thefa/tawing substances would, if
Mobile phase Mix 380 mL of acetonitrile Rand 550 mL of present at a sufficient level, be detected by oneor otherof the tests
waterfor chromawgraphy R and allow to equilibrate; dilute to in the monograph, They are limiud by the general acceptance
1000 mL with waterfor chromawgraphy R and mix. criterion for other/unspecified impurities and/or by the general
Flow rate 1 mIJmin. monograph Substances for pharmaceutical use (2034). It is
Detection Spectrophotometer at 254 nm. therefore not neussary to identify these impurities fQY'
demonstration of compliance, See also 5.10. Control of impuruies
solutions (a), (b), (c) and (d).
in substances for phamzaceuu'cal use) B, C, F, G, H.
Run time 2.5 times the retention time of dexamethasone
acetate.
impurities A and D; use the chromatogram obtained with
reference solution (c) to identify the peak due to impurity E; o
use the chromatogram supplied with dexamethasone tuelate for
peak identification CRS and me chromatogram obtained with
A. 9-fluoro-ll p, 17,21-trihydroxy-I6ct-methylpregna-I,4-
reference solution (d) to identify the peak due to impurity I.
diene-3)2D-dione (dexamethasone),
Relative retention With reference to dexamethasone acetate
= =
(retention time about 22 min): impurity A about 0.4;
= =
impurity D about 0.9; impurity E about 1.2;
impurity I = about 1.4.
- resolution: minimum 3.3 between me peaks due to
impurity D and dexamethasone acetate. o
Limits:
- impun"ty I: not more than 4 times me area of me principal B. 14-fluoro-llP,17-dihydroxy-16IX-methyl-3,20-
peak in the chromatogram obtained with reference dioxopregna-Ld-dien-z l-yl acetate)
- impurity D: not more than 3 times the area of me o 0
H CH,y"\'-JZ
OH CH 3
- impuruies A, E: for each impurity) not more than twice the ····CH3
area of the principal peak in the chromatogram obtained H

o
with reference solution (b) (0"10 per cent);
c. 9-fluoro-11 p, 17-dihydroxy-16IX-methyl-3,20-dioxo-I7a-
pregna-l,4-dien-2l-yl acetate,
(0.5 per cent); o
- disregard limir. 0.5 times the area of the principal peak in "OH
o-JZ CH3
the chromatogram obtained with reference solution (b) '. CH3
(0.05 per cent). ··H

Maximwn 0.5 per cent) determined on 1.000 g by drying in o
vacuo at 105 DC.
D. 9-fluoro-11 p, 17-dihydroxy-16Jl-methyl-3,20-dioxopregna-
ASSAY 1)4-dien-21-yl acetate (betamethasone acetate»
related substances with the following modifications"
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1-748 Dexamethasone Isonicotinate 2022
Dexamethasone Isonicotinate
E. 9-lIuoro-11 ~,17 -dihydroxy-Iee-rnethyl-3,20-dioxopregn-4-

en-21-yl acetate,
497.6 2265-64-7
Action and use

Glucocorticoid.
o /'hE" _
F. 9,11~-epoxy-17-hydroxy-loo-methyl-3,20-dioxo-91l DEFINITION
pregna-l,4-dien-21-yl acetate, 9-Fluoro-11 p, 17-djhydroxy-Iric-methyl-3,20-dioxopregna-
1,4-dien-21-yl pyridine-d-carboxyiate.
o 0
Content
oJ... CH,
CHARACTERS
Appearance
White or almost white crystalline powder.
o
Solublllty
G. 9-lIuoro-llll-hydroxy-16~-methyl-3,20-dioxopregna-I,4- Practically insoluble in water, slightly soluble in anhydrous
dien-21-yl acetate, ethanol and in acetone.
IDENTIFICATION
o Infrared absorption spectrophotometry (2.2.24).
/'-J---'····OH
oJ... CH3 Comparison dexamethasone lsomcotmate CRS.
"CH 3 TESTS
H
o + 142 to + 146 (dried substance).
Suspend 0.200 g in 4.0 mL of ethylacetate R and dilute to
H. 17-hydroxy-1 oo-methyl-3,2Q-dioxopregna-I,4,9(11)-trien- 20.0 mL with ethanol (96 percent,) R. Treat in an ultrasonic
21-yl acetate, bath until a clear solution is obtained.
Related substances
Liquid chromatography (2.2.29). Prepare solutions immediately
before use.
Testsolution Suspend 50.0 mg in 7 mL of acetanitrile Rand
dilute to 10.0 mL with water R. Treat in an ultrasonic bath
until a clear solution is obtained.
o Reference solutian (a) Suspend 5.0 mg of dexamethasone CRS
(impurity A) and 5.0 mg of dexamethasone acetate CRS
I. 9-lIuoro-l7-hydrcxy-I oo-methyl-3,20-dioxopregna-I,4- (impurity B) in ·70 mL of acetanitrile R, add 1.0 mL of the
dlene-Ll p,21-diyl diacetate (dexamethasone 11,21- test solution and dilute to 100.0 mL with water R. Treat in
diacetate). an ultrasonic bath until a clearsolutionis obtained.
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ /'hE" Reference solutian (b) Dilute 1.0 mL of reference solution (a)
Reference solution (c) Suspend 5 mg of dexamethasone
isonicotinate for impurity C identification CRS in 0.7 mL of
acetonitrile R and dilute lO 1 mL with water R. Treat in an
ultrasonic bath until a dear solution is obtained.
Column:
- size: 1= 0.125 m, 0 = 4.0 mm,
- statianary phase: end-capped octade<y/silyl sJ1ica gelfor
chromatography R (5 juri).
Mobile phase:
- mobile phaseA: waterfor chromatography R,
- mobile phaseB: acetonitrile for chromatagraphy R,
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2022 Dexamethasone Sodium Phosphate 1-749

(min) (per cent VlY) (per cent VIJI)
0-2 es 32
2 - 20 68 ---> 50 32 -> 50
Flow rate 1.2 mUmin. o

Detection Spectrophotometer at 240 run. B. 9-ftuoro-11 p, 17-dihydroxy-16~-methyl-3,20-dioxopregna
Injeaion 10 ~L. 1,4-dien-21-yl acetate (dexamethasone acetate),
impurities A and B; use the chromatogram supplied with
dexamethasone isonicotinate for impurity C identification CRS
and the chromatogram obtained with reference solution (c)
to identify the peak due to impurity C.
Relativeretention With reference to dexamethasone o
isonicotinate (retention timc > about 12 min): C. 9-ftuoro-ll p, 17-dihydroxy-16~-methylpregna-I,4-diene
=
impurity A about 0.4; impurity C = about 0.6;
3,20-dione (21-deoxydexamethasone).
=
impurity B about 0.8. _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ PIIE<r
System suitabr7ity Reference solution (a):
impurity B and dexamethasone isonicotinate.
Limits:
- impuniyA: not more than 5 times the area of me Dexamethasone Sodium
corresponding peak in the chromatogram obtained with Phosphate
reference solution (b) (0.5 per cent),
- ;mpun·ty B: not more than 3 times the area of the
reference solution (b) (0.3 per cent),
- impurity C: not more than 3 times the area of the peak
due to dexamethasone isonicotinate in the chromatogram
obtained with reference solution (b) (0.3 per cent),
area of the peak due to dexamethasone isonicotinate in
516.4 2392·39-4
(0.10 per cent),
- LOud: not more than 8 times the area of the peak due to Action and use
dexamethasone isonicotinate in the chromatogram Glucocorticoid.
obtained with reference solution (b) (0.8 per cent),
Preparations
Dexamethasone Sodiwn Phosphate Eye Drops
dexamethasone isonicotinate in the chromatogram
obtained with reference solution (b) (0.05 per cent). Dexamethasone Sodium Phosphate Injection
Loss on drying (2.2.32) Dexamethasone Sodium Phosphate Oral Solution
Maximwn 1.0 per cent, determined on 1.000 g by drying in PIlE" _
°C at a pressure not exceeding 0.1 kPa for 4 h.
vacuo at 105
DEFINITION
ASSAY Disodium 9-ftuoro-ll p, 17-dihydroxy-16~-methyl-3,20
Dissolve DADO g in a mixture of 5 mL of anhydrous formic dioxopregna-l,4-dien-21-yl phosphate.
acid Rand 50 mL of glacial acetic acidR. Titrate with 0.1 M
Content .
perch/one acid, determining the end-point potentiometrically
(2.2.20).
I mL of 0.1 M perchloric acid is equivalent 10 49.76 mg CHARACTERS
of C2S H, 2FNO•. Appearance
White or almost white, very hygroscopic powder.
IMPURITIES
Spea'fied impurities A, B, C. Solubility
Freely soluble in water, slightly soluble in ethanol
o (96 per cent), practically insoluble in methylene chloride.
IDENTIFICATION
o
A. 9-ftuoro-11 p, 17,21-trihydroxy-16~-methylpregna-I,4 Comparison dexamethasone sodium phosphate CRS.
diene-3,20-dione (dexamethasone), If the spectra obtained In the solid state show differences,
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1-750 Dexamethasone Sodium Phosphate 2022
substance separately in the minimum volume of ethanol sodium phosphate CRS in mobile phase A, then dilute to
(96 per cent) R, evaporate to dryness on a water-bam and 100 mL with mobile phase A.
record new spectra using the residues. Reference solution (b) Dissolve 2 mg of dexamethasone sodium
B. Thin-layer chromatography (2.2.27). phosphate for peak identification CRS (containing impurities A,
Test solution Dissolve 10 mg of me substance to be C, D) E, F and G) in mobile phase A and dilute to 2 mL
examined in methanolR and dilute to 10.0 mL with the same with mobile phase A.
solvent. Reference solution (c)
Reference solution Dissolve 10 mg of dexamethasone sodium 100.0 mL with mobile phase A. Dilute 1.0 mL of this
phosphate CRS in methanol R and dilute to 10.0 mL with the solution to 10.0 mL with mobile phase A.
same solvent. Column:
Plate TLC silica gel Fm plate R. - size: f= 0.125 m, 0 = 4.6 mm;
- stationary phase: end-capped octylsily/ silica gelfor
Mobilephase glacialacetic acid R, water R, butanol R
(20:20:60 VIV/V).
chromalography R (5 urn);
Application 5 ~L.
Mobile phase:
Development Over 3/4 of the plate. - mobile phase A: mix 300 mL of solution A and 350 mL of
Drying In air. waterfor chromatography R, adjust to pH 3.8 with acetic
Detection Spray with a solution prepared as follows: dissolve acid R, then add 350 mL of methanolR,
0.25 g of 2,4-dihydroxybenzaldehyde R in gladal acetic acid R, - mobile phase B: adjust 300 mL of solution A to pH 4.0
dilute to 50 mL with the same solvent and add a mixture of with acetic acid R J then add 700 mL of methanol R;
12.5 mL of suiJime acid Rand 37.5 mL of glaciol acetic
acid R, heat the plate at 90 °C for 35 min or until the spots Tim. MobUe phase A Moblle phase B
appear and allow to cool; examine in daylight and in (min) (per cent J'IJI) (per cent I'/JI)
ultraviolet light at 365 DID. 0-3.5 90 to
3.5 - 23.5 90 ....... 60 10 --> 40
23.5 - 34.5 60 ~ 5 40 --> 95
the principal spot in the chromatogram obtained with the 34.5 - 50 5 95
reference solution.
C. Add about 2 mg to 2 mL of sulfuric acidR and shake to Flew rate 1.0 mUmin.
dissolve. Within 5 min, a fa.int yellowish-brown colour Detection Spectrophotometer at 254 om.
develops. Add the solution to 10 mL of water R and mix. Injection 20~.
The colour disappears and a clear solution remains. Identification of impuniies Use the chromatogram supplied
D. Examine the chromatograms obtained in the assay. with dexamethasone sodium phosphate for jJMk
Results The principal peak in the chromatogram obtained identification CRS and the chromatogram obtained with
with the test solution is similar in retention time and size to reference solution (b) to identify the peaks due to
the principal peak in the chromatogram obtained with impurities A, C, D, E, F and Gi use the chromatogram
reference solution (b). obtained with reference solution (a) to identify the peak due
to impurity B.
TESTS
Solution S Relativeretention With reference to dexamethasone sodium
Dissolve 1.0 g in carbon dioxide-free wacer R and dilute to phosphate (retention time = about 22 min):
20 mL with the same solvent. impurity C = about 0.5; impurity D = about 0.6;
impurity E = about 0.8; impurity F = about 0.92;
=
impurity B about 0.95; impurity A about 1.37; =
impurity G = about 1.41.
than reference solution B7 (2.2.2, Method ll).
pH (2.2.3) - resolution: minimum 2.0 between the peaks due to
7.5 to 9.5. impurity B and dexamethasone sodium phosphate.
Dilute 1 mL of solution S to 5 mL with carbon dioxide-free Limits: .
water R. - correction factor. for the calculation of content, multiply the
Specific optical rotation (2.2.7) peak area of impurity A by 0.75;
,+ 75 to + 83 (anhydrous substance). - impurity A: not more than 5 times the area of the
,Dissolve 0

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British Pharmacopoeia 2022

Effective date: I January 2022

London: The Stationery Office

THE DEPARTMENT OF HEALTH AND SOCIAL CARE

Monographs of the European Pharmacopoeia are distinguished by a chaplet

The British Pharmacopoeia Commission has caused this British

The British Pharmacopoeia Commission is appointed, on behalf of the

Members of the British Pharmacopoeia Commission are appointed for a

In order to ensure that the British Pharmacopoeia Commission fulfils its

In addition to the duties listed above, the Chair of the British

Members of Expert Advisory Groups, Panels of Experts and Working

The duties of tbe members are as follows:

Members of the British Pharmacopoeia Commission and its supporting

Chair Professor Kevin M G Taylor BPharm PhD FRPharmS

Vice-Chair Professor Alastair G Davidson BSc PhD FRPharmS

Secretary and Me James Pound BSc

The Commission appointed the following Expert Advisory Groups, Panels

EXPERT ADVISORY GROUPS

HCM: Herbal and M Simmonds (Chair), R Middleton (Vice-Chair), L A Anderson,

MCI: Medicinal A G Davidson (Chair), D Cairns (Vice-Chair), S Bale, H Batchelor,

MC2: Medicinal G Cook (Chair), C T Goddard (Vice-Chair), J Birchall, K Boon, J Cowie, K

MC3: Medicinal M Almond (Chair), J Beach (Vice-Chair), J Beaman, K Foster,

NOM: J K Aronson (Chair), A McFarlane, D Mehta, G P Moss, R Thorpe

PCY: Pharmacy R L Horder (Chair), R Lowe (Vice-Chair), M Ahmed', E Baker, J Beach,

CX: Excipients C Mroz (Vice-Chair), H Batchelor, R Cawthorne, D Deutsch

IGC: Inorganic and C T Goddard (Chair), M Almond, S Boland, P Henrys, G Lay

MIC: Microbiology V Fenton-May (Chair), B Alexander, C Iverson, V Iaitely, J Silva

RAD: Radioactive I Boros, J Brain, D Graham, G Inwards, R D Pickett

VET: Veterinary E Williamson (Chair), A Cairns, S Cockbill, D Evans, E Flahive, B Ward

VIP: Veterinary A M Brady (Chair), R Banks, R Cooney, M Ilort, M Johnson, K Redhead,

ATMP: Advanced J Barry (Chair), E Abranches, C Blue, J Campbell, D Caulfield, R Cowell',

BIO-DPS: P Varley (Chair), A-M Brady (Vice-Chair), C Burns, B Cowper, L Duhau,

Secretariat J Pound (Secretary and Scientific Director)

Administrative N Begum, F Chughtai, B F Delahunty, J Paine, U Rothna

Secondees from the M Francois, R McCoy

I Reydellet (Operations Manager)

A Prince (Business Director)

British Pharmacopoeia 2022

Volumes I and II Medicinal Substances

Implementation dates regarding European Pharmacopoeia publications are

British The BP continues to be part of the Medicines and Healthcare products

regulatory, documentary (BP) and physical (NIESC) standard setting

1 The consultation can befound on lkfollowing webpage: https:llwww.gov.uklgovemmemlamsultauonsl

principles to pharmacopoeial procedures and across the entire Analytical

Revisions A significant number (130, comprising 120 technical revisions and 10

the monographs or other texts on a historical basis, beginning from the 5 th

British The British Pharmacopoeia continues to expand the catalogue of BPCRS

Supplementary Four new Supplementary Chapters to harmonise with the European

SC II A. Changes in Monograph Titles

European Co-operation Agreement

Websites British Pharmacopoeia Website

International Therapeutic Goods Administration, Australia The British

Administration (TGA). The TGA continues to provide advice to British

exchange, contribution to the International Meeting of World

Acknowledgements The British Pharmacopoeia Commission is greatly indebted to the members

The British Pharmacopoeia Commission would also like to thank the

Close co-operation has continued with many organisations in the United

The British Pharmacopoeia Commission wishes to thank the European

The British Pharmacopoeia Commission also acknowledges and appreciates

The British Pharmacopoeia Commission also acknowledges the contribution

Medicinal and Pharmaceutical Substances

Formulated Preparations: General Monographs

Formulated Preparations: Specific Monographs