Eph9th - Suplemento 9.5 - 2018 PDF

EURO PEAN
PHARMACOPOEIA
NINTH EDITION
Supplement 9.5
Published in accordance with the

Convention on the Elaboration of a European Pharmacopoeia
(European Treaty Series No. 50)
~'"
COUNCIL OF EUROPE
*
European Directorate
for the Quality
1 Direction européenne
de la qua lité
of Medicines du médicament
&HealthCare &soins de santé CONSEIL DE l.'.EUROPE
Council of Europe
Strasbourg
The European Pharmacopoeia is published by the European Directorate for the Quality of Medicines &
HealthCare of the Council of Europe (EDQM).
© Council of Europe, 67075 Strasbourg Cedex, France - 2017

All rights reserved. Apart from any fair dealing for the purposes of research or prívate study, this
publication may not be reproduced or transmitted in any form or by any means without the prior
permission in writing of the publisher. Extracts from the European Pharmacopoeia may nevertheless
be used by the subscriber in the context of marketing authorisation procedures, on condition that their
distribution is limited to competent authorities and excludes any third parties.
ISBN: 978-92-871-8331-6
CONTENTS
CONTENTS OF SUPPLEMENT 9.5 xlv
GENERAL CHAPTERS 5533
2. Methods of analysis 5533
2.2. Physical and physicochemical methods 5533
2.2.7. Optical rotation 5535
2.4. Limit tests 5537
2.4.20. Determination of elemental impurities 5539
3. Materials for containers and containers 5543
3.2. Containers 5543
3.2.9. Rubber closures for containers for aqueous parenteral preparations, for powders and for
5545
freeze-dried powders
4. Reagents 5547
4.1.1. Reagents 5549
4.1.3. Buffer solutions 5549
4.2.2. Volumetric solutions 5550
5. General texts 5551
5.4. Residual solvents 5553
5.12. Reference standards 5563
GENERAL MONOGRAPHS 5567
VACCINES FOR HUMAN USE 5581
VACCINES FOR VETERINARY USE 5595
MONOGRAPHS ON SUTURES FOR HUMAN USE 5601
MONOGRAPHS ON SUTURES FOR VETERINARY USE 5607
MONOGRAPHS ON HERBAL DRUGS AND HERBAL DRUG PREPARATIONS 5611
MONOGRAPHS ON HOMOEOPATHIC PREPARATIONS 5619
MONOGRAPHS 5625
INDEX 5761
Note: on the first page of each chapter/section there is a list of contents.

EUROPEAN PHARMACOPOEIA 9.5 Contents of Supplement 9.5
CONTENTS OF SUPPLEMENT 9.5

A vertical line in the margin indicates where part of a text has been revised or corrected. A horizontal line in the margin indicates
where part of a text has been deleted. However, these indications, which are not necessarily exhaustive, are given for information
and do not form an official part of the texts. Editorial changes are not indicated.
Individual copies of texts will not be supplied.
Subscribers to the current version (printed or electronic) of the European Pharmacopoeia have access to an online archive
version of all previous editions of the European Pharmacopoeia.
A list of new reagents published during the course of this edition is available under 'Useful information' in Pharmeuropa Online.
NEWTEXTS
The following texts appear for the first time in the European Pharmacopoeia. They will be implemented on 1 July 2018 at the latest.
MONOGRAPHS Etanercept (2895)
Homoeopathic preparations Fipronil for veterinary use (2869)
Lacosamide (2992)
Acidum succinicum for homoeopathic preparations (2824)
Mometasone furoate monohydrate (2858)
Calcium fluoratum for homoeopathic preparations (2996)
Raltegravir chewable tablets (2939)
Monographs Raltegravir tablets (2938)
Deferiprone (2236) Zolmitriptan (2737)
REVISED TEXTS
The following texts have been technically revised since their last publication. They will be implemented on 1/uly2018.
GENERAL CHAPTERS Sutures for veterinary use
2.2. 7. Optical rotation Polyamide 6 suture, sterile, in distributor for veterinary use
(0609)
2.4.20. Determination of elemental impurities
Polyamide 6/6 suture, steriie, in distributor for veterinary use
3.2.9. Rubber closures for containers for aqueous parenteral (0610)
preparations, for powders and for freeze-dried Poly(ethylene terephthalate) suture, sterile, in distributor for
powders veterinary use (0607)
4. Reagents (chapter 4.1 and new, revised or corrected Herbal drugs and herbal drug preparations
reagents)
Lavender flower (1534)
5.4. Residual solvents
Lavender oil (1338)
5.12. Reference standards Spike lavender oil (2419)
Homoeopathic preparations
MONOGRAPHS Agaricus phalloides for homoeopathic preparations (2290)
General monographs Arsenicum album for homoeopathic preparations (1599)
Pharmaceutical preparations (2619) Aurum chloratum natronatum for homoeopathic preparations
Vaccines for human use (0153) (2141)
Vaccines for veterinary use (0062) Monographs
Acitretin (1385)
Vaccines for human use
Biotin (1073)
Diphtheria, tetanus, pertussis (acellular, component) and
haemophilus type b conjugate vaccine (adsorbed) (1932) Codeine hydrochloride dihydrate (1412)
Codeine monohydrate (0076)
Diphtheria, tetanus, pertussis (acellular, component), hepatitis
B (rDNA), poliomyelitis (inactivated) and haemophilus type Codeine phosphate hemihydrate (0074)
b conjugate vaccine (adsorbed) (2067) Estriol (1203)
Diphtheria, tetanus, pertussis (acellular, component), Folie acid hydrate (0067)
poliomyelitis (inactivated) and haemophilus type b conjugate Gemfibrozil (1694)
vaccine (adsorbed) (2065) Glucosamine hydrochloride (2446)
Diphtheria, tetanus, pertussis (whole cell), poliomyelitis Glucosamine sulfate potassium chloride (2708)
(inactivated) and haemophilus type b conjugate vaccine Glucosamine sulfate sodium chloride (2447)
(adsorbed) (2066) Hydroxypropylcellulose, low-substituted (2083)
Haemophilus type b conjugate vaccine (1219) Hyoscine butylbromide (0737)
Sutures for human use Insulin glargine (2571)
Sutures, sterile non-absorbable (0324) Isoniazid (0146)
xlv
Contents of Supplement 9.5 EUROPEAN PHARMACOPOEIA 9.5
Isotretinoin (1019) Paraffin, liquid (0239)

Lactulose (1230) Pimobendan for veterinary use (2179)
Lactulose, liquid (0924) Polyoxypropylene stearyl ether (2602)
Mometasone furoate (1449)
Sevoflurane(2269)
Neostigmine metilsulfate (0626)
Paraffin, light liquid (0240)
CORRECTED TEXTS
The following texts have been corrected for Supplement 9.5 and specify 'corrected 9.5' above the title. These corrections are to
be taken into account as soon as possible and not later than 28 February 2018 (the end of the month following the month
of publication of Supplement 9.5).
MONOGRAPHS Follitropin concentrated solution (2286)

Vaccines for veterinary use Human coagulation factor Vlla (rDNA) concentrated solution
Foot-and-mouth disease (ruminants) vaccine (inactivated) (2534)
(0063) Hydroxyzine hydrochloride (0916)
Infectious chicken anaemia vaccine (live) (2038) Interferon beta-1 a concentrated solution ( 1639)
Herbal drugs and herbal drug preparations Molgramostim concentrated solution (1641)
Dioscorea nipponica rhizome (2890) Nadroparin calcium (1134)
Monographs Somatropin (0951)
Clomifene citrate (0997) Somatropin concentrated solution (0950)
Follitropin (2285) Somatropin for injection (0952)
TEXTS WHOSE TITLE HAS CHANGED
The titles of the following texts have been changed in Supplement 9.5.
MONOGRAPHS
Monographs
Codeine monohydrate (0076) (previously Codeine)

Folie acid hydrate (0067) (previously Folie acid)
Pimobendan for veterinary use (2179) (previously Pimobendan)
DELETED TEXTS
The following texts are deleted as of 1 /anuary 2018.
MONOGRAPHS
Cholera vaccine (0154)
Cholera vaccine, freeze-dried (0155)
Typhoid vaccine, freeze-dried (0157)
The following text is deleted as of 1 July 2017.
GENERAL CHAPTERS
2.6.19. Test for neurovirulence of poliomyelitis vaccine (oral)
The following text is deleted as of 1 April 2017.
GENERAL CHAPTERS
2.2.60. Melting point - instrumental method
xlvi
EUROPEAN PHARMACOPOEIA 9.5
2.2. Physical and physicochemical

methods
2.2.7. Optical rotation ............................................................ 5535
General Notices (1) apply to ali monographs and other texts 5533
5534 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.5 2.2.7. Optical rotation
07/2018:20207 - a temperature control system that indicates the temperature

with a readability of 0.1 ºC; it may be embedded in the
polarimeter (e.g. a Peltier system) or be an externa! unit
(e.g. a cycle-cryostat), and must be able to maintain
the temperature of the liquid to within ± 0.5 ºC of that
prescribed.
2.2.7. OPTICAL ROTATION
EQUIPMENT PERFORMANCE
PRINCIPLE The accuracy of the scale is checked near the value to be
Optical rotation (also known as optical activity) is the measured or over an appropriate range, usually by means of
property displayed by chiral substances of rotating the plane certified quartz plates. Other certified reference materials may
of polarisation of linearly polarised light. also be suitable (e.g. sucrose solutions).
Optical rotation is considered to be positive (+) for Optical rotation measurements may be used to quantify the
dextrorotatory substances (i.e. those that rotate the plane amount of an enantiomer or the ratio of enantiomers present
of polarisation in a clockwise direction when viewed in the in a sample. For that purpose, the linearity must be checked,
direction facing the oncoming light beam) and negative (-) for example using sucrose solutions.
for laevorotatory substances (i.e. anticlockwise rotation). PROCEDURE
The angle of optical rotation a of a liquid is the angle of Determine the zero of the polarimeter and the angle of rotation
rotation of the plane of polarisation, expressed in degrees (º), of the liquid at a wavelength of 589 nm and a temperature
at the wavelength of the D-line of sodium (A= 589.3 nm) of 20 ± 0.5 ºC, unless otherwise prescribed. The zero of the
measured at 20 ºC through the liquid when using a path polarimeter is determined with the sample cell closed.
length of LOO dm. For neat liquids, the zero is determined with an empty sample
The specific optical rotation [a]~º of a substance in solution is cell.
calculated from the angle of optical rotation, as defined above, For solutions, the zero is determined with the sample cell
with reference to a path length of 1.00 dm and a concentration filled with the same solvent as that used for the solution to be
of the substance to be examined of 1 g/mL. The specific optical examined and measured at the same temperature. The sample
rotation of a substance in solution is always expressed with preparation procedure is prescribed in the monograph.
reference to a given solvent and concentration. Calculate the specific optical rotation at temperature t and
As sorne equipment may not use sodium lamps, the wavelength wavelength Á using the following formulae.
of measurement is given as 589 nm instead of 589.3 nm. For neat liquids, the density of the liquid is taken into account:
In certain cases specified in the monograph, the angle of t a
optical rotation is measured at other temperatures, other [ah= l·pt
wavelengths and/or in cells with a path length other than
1.00 dm. For solutions:
t _ lOOOa
In the conventional system adopted by the Pharmacopoeia, the [ ª>-
] -
specific optical rotation is expressed by its value without units; l·c
the actual units, degree millilitres per decimetre gram
[(º)·ml·dm- 1·g- 1] are understood. a angle of rotation measured at temperature t
and wavelength Á, in degrees;
EQUIPMENT path length of the polarimeter sample cell, in
The polarimeter typically consists of: decimetres;
a light source, for example a sodium discharge lamp, a Pt density determined at the temperature of
light-emitting dio de (LED) or another light source capable measurement t, in grams per cubic centimetre;
of providing radiation at the desired wavelength (589 nm for the purposes of the Pharmacopoeia, density
unless otherwise prescribed in the monograph); if the light is replaced by relative density (2.2.5);
source is polychromatic, a means of isolating the required e concentration of the solution, in grams per
wavelength is necessary, e.g. an optical filter; litre.
- a polariser and an analyser;
- a sample cell with a path length of 1.00 dm, unless otherwise When the limits for optical rotation or specific optical rotation
specified in the monograph; are expressed as the dried substance, the anhydrous substance
- a detection system to measure the angle of optical rotation, or the solvent-free substance, the result must be corrected for
which must be capable of giving readings to at least the loss on drying (2.2.32), water content (2.5.12 or 2.5.32) or
nearest 0.01 º, unless otherwise specified in the monograph; content of solvent as appropriate.

2.4. Limit tests

2.4.20. Determination of elemental impurities ................... 5539

EUROPEAN PHARMACOPOEIA 9.5 2.4.20. Determination of elemental impurities
07/2018:20420 peroxide, at various concentrations, can be used to dissolve

the samples. The viscosity of sulfuric acid is greater than that
of the other acids and is to be taken into account as it can
affect the overall fluidity of the solution.
The choice of solvents also includes, but is not limited to,
2.4.20. DETERMINATION OF the use of dilute bases, straight or diluted organic solvents,
combinations of acids or bases, and combinations of organic
ELEMENTAL IMPURITIES solvents.
INTRODUCTION Acids, bases, and hydrogen peroxide of high purity must be
This chapter describes the general approach for the used, especially when ICP-MS is employed. For aqueous
determination of elemental impurities in medicinal products solutions, use deionised distilled water R. Diluents must
or substances for pharmaceutical use. As the chemical be checked for interference if they are used in an analysis.
composition of the considered samples and the specification Because it is not always possible to obtain organic solvents
limits for the element(s) of interest vary considerably, it is that are free from elemental impurities, organic solvents of
not possible to describe all suitable sample preparation and the highest purity possible with regard to these contaminants
measurement methods. Therefore, any method that fulfils the must be used. Specifically for ICP techniques, where samples
requirements described in this chapter may be used. are introduced into the plasma via solution nebulisation,
it is important to consider the potential matrix effects and
The results of the analysis are acceptable only if the system interferences that might arise from the solvent. The use of an
suitability has been demonstrated by a suitable test. Before appropriate interna! standard and/or matching the standard
the initial use of a method, the analyst must ensure that the matrix with samples should be applied for ICP-AES and
method is appropriate for the samples and instruments used. ICP-MS analyses in cases where accuracy and precision are
This is accomplished by applying a validation procedure to not sufficient. In any case, the selection of an appropriate
methods not described in the individual monograph or by interna! standard should take into account the element(s) of
a system suitability test for methods which are described in interest, ionisation energy, wavelengths or masses, and the
the monograph. Decision trees for the choice of the sample nature of the sample matrix.
preparation and the measurement procedures are presented in
Figures 2.4.20.-1 and 2.4.20.-2. Where a sample is found not to be soluble in any acceptable
solvent, a variety of digestion or incineration techniques can
PROCEDURES be employed. These include hot-plate digestion, incineration
and microwave-assisted digestions, using an open- or
As a reference procedure is not provided for each element,
closed-vessel.
matrix and concentration, the choice of procedure according
to Figures 2.4.20.-1 and 2.4.20.-2, including sample The decision regarding the type of digestion technique to be
preparation, detection technique and instrument parameters, used depends on the nature of the sample being digested, as
is the responsibility of the user. well as on the element(s) of interest and the concentration
Use the flow chart in Figure 2.4.20.-1 to define the sample range of the elements to be quantified. Open-vessel digestion
preparation method and the flow chart in Figure 2.4.20.-2 to is not recommended for the analysis of volatile elements.
define the measurement method. The sample preparation The suitability of a digestion technique, whether open-
method shouid yield a sufficient quantity of sample to allow or closed-vessel, should be supported by spike recovery
quantification of each element at the specified limit stated in experiments in order to verify that, within an acceptable
the individual monograph or the general chapter. tolerance, volatile elements have not been lost during sample
preparation. The digestion cycle is suitable if a clear solution
All suitable sample preparation methods and measurement is obtained.
techniques (e.g. 2.2.22. Atomic emission spectrometry (AES),
2.2.23. Atomic absorption spectrometry (AAS), 2.2.37. X-ray It is important to consider the selection of the type, the
fluorescence spectrometry (XRFS), 2.2.57. Inductively material of construction, the pretreatment, and the cleaning
coupled plasma-atomic emission spectrometry (ICP-AES), of analytical labware used in elemental analyses. The material
2.2.58. Inductively coupled plasma-mass spectrometry must be inert and, depending on the specific application,
(ICP-MS), 2.4.2. Arsenic, 2.4.8. Heavy metals, 2.4.9. Iron, resistant to caustics, acids, and/or organic solvents. For sorne
2.4.10. Lead in sugars, 2.4.15. Nickel in polyols, 2.4.31. Nickel in analyses, care must be exercised to prevent the adsorption of
hydrogenated vegetable oils) can be used for the determination elemental impurities onto the surface of a vessel, particularly
of elemental impurities, if the method has been verified before in ultra-trace analyses. Contamination of sample solutions by
the initial use by a system suitability test or a validation elemental impurities and ions present in the container can
procedure according to this chapter. also lead to inaccurate results.
If no sample preparation and/or measurement method is The use of volumetric glassware that <loes not comply
described in the individual monograph, a suitable sample with Class A requirements of the appropriate International
preparation and/or measurement method must be developed Standard of the International Organization for Standardization
and validated (see Figures 2.4.20.-1 and 2.4.20.-2). (ISO) is acceptable if the validation or the system suitability
test of the method using such glassware have experimentally
SAMPLE PREPARATION demonstrated that the method is suitable for the intended
Sample preparation is critica! to the success of elemental purpose.
analysis. Many techniques not using direct measurement are CAUTION: when using high-pressure digestion vessels and
heavily dependent on sample transport. microwave laboratory equipment, the safety precautions and
If an atomisation system is used, the most conventional means operating instructions given by the manufacturer must be
by which samples are introduced into the atomisation system followed.
is by solution nebulisation. In this case, solid samples must
be dissolved in order to be introduced into the atomisation MEASUREMENT
system. Samples may be dissolved in any appropriate solvent. Method. The choice of the techniques depends mainly on
The use of aqueous or dilute nitric acid solutions is strongly the sample matrix and the characteristics and specification
recommended, due to minimal interference with these limits of the element(s) of interest. Analyse according to the
solvents compared to other solvents. Hydrochloric acid, instructions of the manufacturer of the equipment regarding
hydrofluoric acid, perchloric acid, sulfuric acid and hydrogen program and wavelength.
General Notices (1) apply to all monographs and other texts 5539
2.4.20. Determination of elemental impurities EUROPEAN PHARMACOPOEIA 9.5
No Yes
No
No
Yes
No
Yes Yes
Yes
No
Yes Perform closed- V erify procedure

vessel digestion and/or instrument
Perform open- or closed-

No
vessel digestion or
incineration
No
Figure 2.4.20.-2
Figure 2.4.20.-1. - Elemental impurities decision tree: sample preparation
System suitability. A system suitability test must be carried Cakulation. The blank value of reagents must be taken into
out on the day of the analysis to ensure that the sample account for the calculation of the content. Upon completion
preparation and measurement system are appropriate. of the analysis, the concentration of a given element in
Acceptance criterion for preparation of sample solution: a clear the sample is calculated by the software of the instrument
solution is obtained. from the concentration of the element in the test solution.
If no calculation software is available or no indication for
Acceptance criterion for measurement system: the measured calculation is given in the general chapter corresponding to
concentration of a standard solution of the element at a
concentration within the range of the used calibration curve
<loes not differ from the actual concentration by more than
20 per cent.

EUROPEAN PHARMACOPOEIA 9.5 2.4.20. Determination of elemental impurities
Figure 2.4.20.-1:
prepare the sample
No
Develop a
test procedure
Yes
No
Validate the
test procedure
Perform the Verify the

analysis using measurement
monograph procedure system
Perform analysis using

the validated procedure
Yes No
Verify the Yes

measurement
system
Report result
Figure 2.4.20.-2. - Elemental impurities decision tree: measurement
the method used, the concentration of a given element in matrix and instrument used. This is accomplished by
the sample can be calculated from the concentration of the following the validation procedure before the initial use and
element in the solution using the following expression: the system suitability test on the day of the analysis.
For elemental impurities, validation of a limit test must
e= Ax Vi x v2 include specificity and limit of detection.
m V3
The following section defines the characteristics for the
C concentration of element in the analysed sample, acceptability of a quantitative procedure. It must be
in micrograms per gram; demonstrated experimentally that such a procedure complies
with the validation requirements, with an appropriate system
A instrument reading of the concentration of the suitability test using material spiked with a suitable reference
element in the sample solution, in micrograms per material. The test materials must be spiked before any sample
millilitre; preparation steps. For example, if a test material is to be
m mass of the sample in the initial sample solution, digested, the material must be spiked at the beginning of the
in grams; digestion procedure.
V1 volume of the initial sample preparation, in SPECIFICITY
millilitres; Specificity is the ability to ensure that the analytical procedure
v2 total volume of any dilution performed, in (sample preparation and measurement) allows a reliable
millilitres; determination of the element(s) of interest in the presence of
V3 volume of initial sample preparation used in any components (e.g. carrier gas, impurities, matrix) that may be
dilution performed, in millilitres. expected to be present.
Acceptance criteria: the procedure must be able to assess
unequivocally each elemental impurity to be determined with
VALIDATION REQUIREMENTS this procedure in the presence of components that may be
expected to be present, including other elemental impurities,
Sorne validation requirements provided below may differ from matrix components and other sources of interference;
those provided in general chapters of the Ph. Eur. (e.g. 2.2.22 specificity is demonstrated by complying with the accuracy
(AES), 2.2.23 (AAS), 2.2.57 (ICP-AES), 2.2.58 (ICP-MS)). requirement for the element(s) to be determined.
Before the initial use of the selected procedure, the analyst RANGE
must ensure that the sample preparation and measurement Acceptance criterion: range is demonstrated by complying
method are appropriate for the element(s) of interest, sample with the recovery requirement.
2.4.20. Determination of elemental impurities EUROPEAN PHARMACOPOEIA 9.5
ACCURACY INTERMEDIA TE PRECISION

Verify the accuracy using a certified reference material The effect of random events (intra-laboratory variations) on
(elemental impurity solutions CRS may be used) or by the analytical precision of the method must be established.
performing a test for recovery. Acceptable experiments for establishing intermediate
precision include performing the repeatability analysis on
The recovery may be determined on a sample of the substance different days, or with different instrumentation, or by
to be examined, spiked with a known quantity of a reference different analysts. Only 1 of the 3 experiments is required to
standard of the element of interest (3 concentration levels in demonstrate intermediate precision.
the range of 50-150 per cent of the intended specification limit, Acceptance criterion: the relative standard deviation is not
even if the original concentration of the reference standard is more than 25 per cent.
at the specified value), in triplicate.
LIMIT OF QUANTIFICATION
Use the results from the accuracy study. Determine the lowest
Acceptance criterion: spike recovery is within 70 per cent and
concentration meeting the acceptance criterion.
150 per cent for the mean of 3 replicates at each concentration.
Acceptance criterion : the limit of quantification is below the
REPEATABILITY specification limit.
Test samples: either 6 independent samples of the substance LIMIT OF DETECTION (ONLY APPLICABLE TO LIMIT
to be examined spiked with a suitable reference standard at TESTS)
the specified concentration level, or 3 concentration levels Determine the lowest concentration giving a signal clearly
prepared in triplicate. distinct from that obtained with a blank solution.
Acceptance criterion: the relative standard deviation is in both Acceptance criterion: the limit of detection is not more than
cases not more than 20 per cent. 0.5 times the concentration of the specification limit.

3.2. Containers
3.2.9. Rubber closures for containers for aqueous
parenteral preparations, for powders and for
freeze-dried powders ............................................................ 5545

EUROPEAN PHARMACOPOEIA 9.5 3.2.9. Rubber closures for containers
07/2018:30209 CHARACTERS
Rubber closures are elastic. They are translucent or opaque
and have no characteristic colour, the latter depending
on the additives used. They are practically insoluble in
tetrahydrofuran, in which, however, a considerable reversible
swelling may occur. They are homogeneous and practically
3.2.9. RUBBER CLOSURES FOR free from flash and adventitious materials (for example, fibres,
CONTAINERS FOR AQUEOUS foreign particles, waste rubber).
PARENTERAL PREPARATIONS, IDENTIFICATION
FOR POWDERS AND FOR Identification of the type of rubber used for the closures is not
FREEZE-DRIED POWDERS within the scope of this specification. The identification tests
given below distinguish between closures made from rubber
Rubber closures for containers for aqueous parenteral and those made from silicone elastomer and plastic materials
preparations, for powders and for freeze-dried powders are but do not differentiate ali types of rubber. Other identity
elastomers made of materials obtained by vulcanisation tests may be carried out with the aim of detecting differences
(cross-linking) of macromolecular organic substances, using in a batch compared with the closures used for compatibility
appropriate additives. They cover all types of rubber closures testing. One or more of the following analytical methods
including stoppers for vials, sealing disks and plunger stoppers may be applied for this purpose: determination of relative
for cartridges, as well as rubber tip caps, needle shields and density, determination of sulfated ash, determination of sulfur
plunger stoppers for syringes. The elastomers are polymers, content, thin-layer chromatography carried out on an extract,
obtained by chemical synthesis, or are polymers of natural ultraviolet absorption spectrophotometry of an extract, infrared
origin. The choice of the principal components and of the absorption spectrophotometry of a pyrolysate or attenuated
various additives (for example, vulcanisers, accelerators, total reflectance (ATR).
stabilisers, pigments) depends on the properties required A. Infrared absorption spectrophotometry (2.2.24). Examine
for the finished article. The specifications apply to rubber by attenuated total reflectance (ATR).
closures made from rubber of 1 kind only, to coated closures, If necessary, cut the sample along an appropriate axis
to bi-layer seals and to lubricated closures. Coated closures and examine the cut surface. For coated, bi-layer and
consist of a bulk of rubber, bearing on its surface or part of lubricated closures, perform the test for each different part
its surface a layer of a different polymer. Bi-layer seals are of the closure. This is not required for silicone oil used as
composed of 2 different layers of rubber, 1 of which exhibits lubricant. Identification of silicone oil can be performed
a higher level of chemical purity and is intended for contact prior to it being used.
with a pharmaceutical preparation; the other layer exhibits a
Comparison: type sample.
higher level of elasticity and is intended to improve self-sealing
and fragmentation resistance of the seal. Lubricated rubber If direct ATR measurement on the surface is not feasible
closures are closures treated with silicone oil (3.1.8) or other (mainly rubber closures filled with carbon black), heat an
lubricants, for example materials chemically or mechanically appropriate amount of rubber in a heat-resistant test-tube
bonded to the closures. over an open flame to dry the sample and continue heating
until pyrolysate vapours are condensed near the top edge
If closures are lubricated they comply in lubricated state of the test-tube. Examine the pyrolysate of the sample by
with the requirements as defined in this general chapter. ATR and compare the spectrum with that obtained with
The specifications do not apply to closures made from the pyrolysate of the type sample.
silicone elastomer (which are dealt with in general chapter B. Total ash (2.4.16).
3.1.9. Silicone elastomer for closures and tubing).
If the sample has not been subjected to steam sterilisation,
Rubber closures may be classified in 2 types: type I closures drying at 100-105 ºC can be omitted. Determine the
meet the strictest requirements and are preferred; type II percentage content of total ash in the sample to be
closures have mechanical properties suitable for special examined and compare with the percentage content of total
uses (for example, multiple piercing) and cannot meet ash in the type sample (A 0). The total ash content falls
requirements as severe as for type I closures because of their within the following ranges depending on the total ash
chemical composition. content of the type sample, or, if not available, in the range
The closures chosen for use with a particular preparation are defined as the target for the specific rubber type.
such that: Total ash in the type sample, Limit for total ash in the
A0 (per cent) sample (per cent)
- the components of the preparation in contact with the
A0 :::; 5.0 (A 0 - 0.75) to (A 0 + 0.75)
closures are not adsorbed onto the surface of the closures
and do not migrate into or through the closures to an 5.0 < A 0 :::; 10 (A 0 - 1.0) to (A 0 + 1.0)
extent suf:ficient to affect the preparation adversely;
A 0 > 10 (A 0 - 2.0) to (A 0 + 2.0)
- the closures do not release substances in quantities
sufficient to affect the stability of the preparation or to In addition to the use of platinum and silica crucibles
presenta risk of toxicity; described in general chapter 2.4.16, porcelain crucibles may
be used. The sample may be ignited using a microwave
- the closures are compatible with the preparation for which
oven instead of a muffle furnace.
-
they are used throughout its period of validity.
The manufacturer of the preparation must obtain from the TESTS
supplier an assurance that the composition of the closure Solution S. Place a number of uncut closures with a total
<loes not vary and that it is identical to that of the closure surface area of about 100 cm 2 in a wide-necked flask (type
used during compatibility testing. If the supplier informs the I glass, 3.2.1), add 200 mL of water R and weigh. Cover the
manufacturer of the preparation that changes have been made
mouth of the flask with a borosilicate-glass beaker. Heat in
to the composition, a risk assessment should be applied to
an autoclave so that a temperature of 121 ± 2 ºC is reached
determine the need to repeat the compatibility testing, totally
within 20-30 min and maintain at this temperature for
or partly, depending on the nature of the changes. 30 min. Immerse the temperature probe for the autoclave
The closures are washed and may be sterilised before use. programme-control in water in a container comparable to that
3.2.9. Rubber dosures for containers EUROPEAN PHARMACOPOEIA 9.5
used for the sample. Cool to room temperature over about Residue on evaporation. Evaporate 50.0 mL of solution S to
30 min. Make up to the original mass with water R. Shake dryness on a water-bath and dry at 100-105 ºC. The residue
and decant the solution immediately. Shake solution S before weighs not more than 2.0 mg for type I closures and not more
each test. If using a tightly closed flask (type I glass, 3.2.1) than 4.0 mg for type II closures.
with an inert closure inste ad of a wide-necked flask covered Volatile sulfides. Place closures, cut if necessary, with a total
with a borosilicate-glass beaker, it is not necessary to make surface area of 20 ± 2 cm 2 in a 100 mL conical flask and add
up to the original mass. 50 mL of a 20 g/L solution of citric acid monohydrate R. Place
Blank solution. Prepare a blank solution in the same manner a piece of lead acetate paper R over the mouth of the flask and
using 200 mL of water R. maintain the paper in position by placing over it an inverted
Appearance of solution S. Solution S is not more intensely weighing bottle. Heat in an autoclave at 121 ± 2 ºC for 30 min.
coloured than reference solution GY5 (2.2.2, Method JI). For Any black stain on the paper is not more intense than that of
type I closures, solution S is not more opalescent than reference a standard, treated in the same manner, prepared by mixing
suspension II (2.2.1) and for type II closures, solution S is 50 mL of a 20 g/L solution of citric acid monohydrate R and
not more opalescent than reference suspension III. In case of 5.0 mL of a freshly prepared 0.0308 g/L solution of sodium
nephelometric determination, the limit for type I closures is sulfide R.
6 NTU and the limit for type II closures is 18 NTU.
The tests for penetrability, fragmentation and self-sealing are
Acidity or alkalinity. To 20 mL of solution S add 0.1 mL of performed on whole closures.
bromothymol blue solution Rl. Carry out a titration using
20.0 mL of the blank (see solution S). Not more than 0.3 mL Por the tests for penetrability, fragmentation and self-sealing,
of 0.01 M sodium hydroxide or 0.8 mL of 0.01 M hydrochloric treat non-sterilised closures as described for the preparation
acid is required to change the colour of the indicator to blue or of solution S and allow to dry. To perform these 3 tests, use
yellow, respectively. If after adding the indicator the solution for each closure a new, lubricated, long-bevelW (bevel angle
is green, it is neutral and no titration is needed. 12 ± 2º) hypodermic needle with an external diameter of
0.8 mm and pierce the closures with the needle perpendicular
Absorbance. Carry out the test within 5 h of preparation
to the surface without rotating the needle.
of solution S. Filter solution S through a membrane filter
(nominal pore size 0.45 µm), rejecting the first few millilitres Penetrability. For closures intended to be pierced by a
of filtrate. Measure the absorbance (2.2.25) of the filtrate at hypodermic needle, carry out the following test. Fill 10
wavelengths from 220-360 nm using the blank (see solution S) suitable vials to the nominal volume with water R, fit the
as compensation liquid: absorbance within the 220-360 nm closures to be examined and secure with a cap. The force
range <loes not exceed 0.2 for type I closures or 4.0 for type II required for piercing, determined with an accuracy of ± 0.25
closures. If necessary, dilute the filtrate before measurement N, is not greater than 10 N for each closure.
of the absorbance and correct the result for the dilution. Fragmentation. Por closures intended to be pierced by a
Reducing substances. Carry out the test within 4 h of hypodermic needle, carry out the following test. If the closures
preparation of solution S. To 20.0 mL of solution S add are to be used for aqueous preparations, introduce in 12
1 mL of dilute sulfuric acid R and 20.0 mL of 0.002 M clean vials a volume of water R corresponding to the nominal
potassium permanganate. Boil for 3 min. Cool. Add 1 g volume minus 4 mL, close the vials with the closures to be
of potassium iodide R and titrate immediately with 0.01 M examined, secure with a cap and allow to stand for 16 h. If the
sodium thiosulfate, using 0.25 mL of starch solution R as closures are to be used with dry preparations, close 12 clean
indicator. Carry out a titration using 20.0 mL of the blank vials with the closures to be examined. Using a needle fitted to
(see solution S). The difference between the titration volumes a clean syringe, inject into the vial 1 mL of water R and remove
is not greater than 3.0 mL for type I closures and 7.0 mL for 1 mL of air; carry out this operation 4 times for each closure,
type II closures. piercing the closure each time at a different site. Use a new
Ammonium (2.4.1, Method A): maximum 2 ppm. needle for each closure and check that the needle is not blunted
Dilute 5 mL of solution S to 14 mL with water R. during the test. Pass the liquid in the vials through a filter with
a pore size of 0.5 µm. Count the fragments of rubber visible
Extractable zinc: maximum 5 µg of extractable Zn per to the naked eye. The total number of fragments <loes not
millilitre of solution S. exceed 5. This limit is based on the assumption that fragments
Atomic absorption spectrometry (2.2.23, Method I). with a diameter equal to or greater than 50 µm are visible to
Test solution. Use solution S. If results are outside the the naked eye; in cases of doubt or dispute, the fragments are
calibration range, dilute 1O.O mL of solution S to an examined with a microscope to verify their nature and size.
appropriate volume with 0.1 M hydrochloric acid. Self-sealing test. For closures intended to be used with
Reference solutions. Prepare the reference solutions using multidose containers, carry out the following test. Fill 10 vials
zinc standard solution (10 ppm Zn) R diluted with 0.1 M matching the design of the stopper to the nominal volume
hydrochloric acid. with water R, fit the closures to be examined, secure with a
Source: zinc hollow-cathode lamp. cap and crimp tightly. Pierce each closure 1O times, piercing
Wavelength: 213.9 nm. the closure each time at a different site. Immerse the vials
upright in a 1 g/L solution of methylene blue R and reduce the
Atomisation device: air-acetylene flame.
external pressure by 27 kPa for 10 min. Restore atmospheric
Extractable heavy metals (2.4.8): maximum 2 ppm. pressure and leave the vials immersed for 30 min. Rinse the
Solution S complies with test A. Prepare the reference solution outside of the vials. None of the vials contains any trace of
using lead standard solution (2 ppm Pb) R. coloured solution.
( 1) See ISO 7864, Sterile hypodermic needles for single use.

4. Reagents
4.1. Reagents, standard solutions, buffer solutions ............ 5549 4.1.3. Buffer solutions ............................................................ 5549
4.1.1. Reagents ........................................................................ 5549 4.2.2. Volumetric solutions .................................................... 5550

EUROPEAN PHARMACOPOEIA 9.5 4.1.3. Buffer solutions
07/2018:40100 mp: 161 ºC to 162 ºC.

Nicotinoyl hydrazide. C6 H 7Np. (Mr 137.1). 1202400.
[553-53-7]. Pyridine-3-carbohydrazide.
White or almost white powder or crystalline powder, soluble
4.1. REAGENTS, STANDARD in water.
mp: about 160 ºC.
SOLUTIONS, BUFFER SOLUTIONS
Organosilica polymer, multi-layered, octadecylsilyl,
Where the name of a substance or a solution is followed by the
end-capped. 1202500.
letter R (the whole in italics), this indicates a reagent included
in the following list. The specifications given for reagents do Synthetic, spherical hybrid particles, multi-layered, containing
not necessarily guarantee their quality for use in medicines. both inorganic (silica) and organic (organosiloxanes)
components, chemically modified at the surface by the
Within the description of each reagent there is a 7-digit
bonding of octadecylsilyl groups. To minimise any interaction
reference code in italics (for example, 1002501). This number,
with basic compounds, they are carefully end-capped to cover
which will remain unchanged for a given reagent during
most of the remaining silanol groups.
subsequent revisions of the list, is used for identification
purposes by the Secretariat, and users of the Pharmacopoeia Raltegravir potassium. C20H 20 FKNp 5 • 1202600.
may also find it useful, for example in the management of [871038-72-1].
reagent stocks. The description may also include a CAS
See Raltegravir potassium (2887).
number (Chemical Abstract Service Registry Number)
recognisable by its typical format, for example 9002-93-1. Resin for hydrophobic interaction chromatography.
Sorne of the reagents included in the list are toxic and are to be 1202700.
handled in conformity with good quality control laboratory Non-porous resin consisting of spherical polymethacrylate
practice. particles bonded with butyl groups.
Reagents in aqueous solution are prepared using water R. pH limits of use: 2 to 12.
For liquid chromatography, water for chromatography R is
used for the preparation of mobile phases when water, or an Silica gel for chromatography, butylsilyl. 1076200.
aqueous solution, is one of the components. Where a reagent A very finely divided silica gel, chemically modified at the
solution is described using an expression such as 'hydrochloric surface by the bonding of butylsilyl groups.
acid (10 g/L HCl)', the solution is prepared by an appropriate
dilution with water R of a more concentrated reagent solution Silica gel for chromatography (hybrid material),
specified in this chapter. Reagent solutions used in the limit octadecylsilyl, ethylene-bridged, charged surface,
tests for barium, calcium and sulfates are prepared using end-capped. 1202800.
distilled water R. Where the name of the solvent is not stated, Synthetic, spherical ethylene-bridged hybrid particles with a
an aqueous solution is intended. charged surface, containing both inorganic (silica) and organic
The reagents and reagent solutions are to be stored in (organosiloxanes) components, chemically modified at the
well-closed containers. The labelling should comply with the surface by the bonding of octadecylsilyl groups. To minimise
relevant national legislation and international agreements. any interaction with basic compounds they are carefully
end-capped to cover most of the remaining silanol groups.
Silica gel for chromatography, octadecylsilyl, for separation
07/2018:40101 of polycydic aromatic hydrocarbons. 1202900.
A very finely divided ultrapure silica gel, chemically modified
at the surface by the bonding of octadecylsilyl groups,
optimised for the analysis of polycyclic aromatic hydrocarbons.
4.1.1. REAGENTS
Anhydrous colloidal silica. 1202000. [7631-86-9]. 07/2018:40103
See Anhydrous colloidal silica (0434).
l ,2-Diamino-4,5-methylenedioxybenzene dihydro-
chloride. C7 H 10Cl2 Np 2 • (Mr 225.1). 1202100. [81864-15-5].
2H- l ,3-Benzodioxole-5,6-diamine dihydrochloride.
Content: minimum 99 per cent (HPLC).
4.1.3. BUFFER SOLUTIONS
0.25 M Sodium phosphate buffer solution pH 7.5. 4016100.
Hydrazine sulfate. H 6Np 4S. (Mr 130.1). 1043400.
[10034-93-2]. Dissolve 3.90 g of sodium dihydrogen phosphate R in 70 mL
of water R, adjust to pH 7.5 with a 300 g/L solution of sodium
Colourless crystals, sparingly soluble in cold water, soluble
hydroxide R and dilute to 100 mL with water R.
in hot water (SO ºC) and freely soluble in boiling water,
practically insoluble in ethanol (96 per cent). 0.1 M Tris-hydrochloride buffer solution pH 7.5. 4016200.
Content: minimum 99 per cent. Dissolve 3.03 g of tris(hydroxymethyl)aminomethane R in
200 mL of water R, adjust to pH 7.5 with hydrochloric acid R
Isonicotinic acid. C6 H 5N0 2 • (Mr 123.1). 1202200. [55-22-1].
and dilute to 250 mL with water R.
Pyridine-4-carboxylic acid.
Creamish-white powder, sparingly soluble in water. Guanidine-tris(hydroxymethyl)aminomethane buffer
mp: about 311 ºC. solution pH 8.3. 4016300.
Dissolve 1.21 g of tris(hydroxymethyl)aminomethane R in
Maltol. C6 Hp 3 • (Mr 126.1). 1202300. [118-71-8]. 87.5 mL of a 764 g/L solution of guanidine hydrochloride R.
3-Hydroxy-2-methyl-4H-pyran-4-one. Adjust to pH 8.3 with hydrochloric acid R and dilute to 100 mL
White or almost white crystalline powder, soluble in hot water. with water R.
4.2.2. Volumetric solutions EUROPEAN PHARMACOPOEIA 9.5
07/2018:40202 1 mL of 0.1 M ammonium and cerium sulfate is equivalent to

38.21 mg of Fe(C 2 H 10N 2 )(S04 ) 2,4HzÜ.
Dilution. Use a diluted solution of sulfuric acid R (59 g/L
H 2SO 4 ) while cooling the solution.
4.2.2. VOLUMETRIC SOLUTIONS 0.1 M Cerium sulfate. 3001100.

Dissolve 40.4 g of cerium sulfate R in a mixture of 500 mL of
0.1 M Ammonium and cerium nitrate. 30001 OO. water R and 50 mL of sulfuric acid R. Allow to cool and dilute
Shake for 2 min a solution containing 56 mL of sulfuric acid R to 1000.0 mL with water R.
and 54.82 g of ammonium and cerium nitrate R, then add 5 Standardisation. Dissolve 0.300 g of ferrous ethylenediam-
successive quantities, each of 100 mL, of water R, shaking after monium sulfate RV in 50 mL of a diluted solution of sulfuric
each addition. Dilute the clear solution to 1000.0 mL with acid R (49 g/L H 2 SO 4 ). Titrate with the cerium sulfate solution,
water R. Standardise the solution after 1O days. determining the end-point potentiometrically (2.2.20) or
Standardisation. Dissolve 0.300 g of ferrous ethylenediam- using 0.1 mL of ferroin R as indicator.
monium sulfate RV in 50 mL of a diluted solution of sulfuric 1 mL of 0.1 M cerium sulfate is equivalent to 38.21 mg of
acid R (49 g/L H 2S0 4 ). Titrate with the ammonium and cerium Fe(C 2H 10N 2 )(S0 4 )2'4H 20.
nitrate solution, determining the end-point potentiometrically
(2.2.20) or using 0.1 mL of ferro in R as indicator. 0.02 M Potassium permanganate. 3005300.
1 mL of 0.1 M ammonium and cerium nitrate is equivalent to Dissolve 3.2 g of potassium permanganate R in water R and
38.21 mg of Fe(C2 H 10N 2 )(S0 4 )2'4HzÜ. dilute to 1000.0 mL with the same solvent. Heat the solution
for 1 h on a water-bath, allow to cool and filter through a
Storage: protected from light.
sintered-glass filter (2.1.2).
0.1 M Ammonium and cerium sulfate. 3000300. Standardisation. Dissolve 0.300 g of ferrous ethylenediammo-
Dissolve 65.0 g of ammonium and cerium sulfate R in a nium sulfate RV in 50 mL of a diluted solution of sulfuric acid R
mixture of 500 mL of water R and 30 mL of sulfuric acid R. (49 g/L H 2 S0 4 ). Titrate with the potassium permanganate
Allow to cool and dilute to 1000.0 mL with water R. solution, determining the end-point potentiometrically
(2.2.20) or using 0.1 mL of ferro in R as indicator. Standardise
Standardisation. Dissolve 0.300 g of ferrous ethylenediam- immediately before use.
monium sulfate RV in 50 mL of a diluted solution of sulfuric
acid R (49 g/L H 2S0 4). Titrate with the ammonium and cerium 1 mL of 0.02 M potassium permanganate is equivalent to
sulfate solution, determining the end-point potentiometrically 38.21 mg of Fe(C 2H 10N2)(S0 4 ) 2,4HzO.
(2.2.20) or using 0.1 mL of ferroin R as indicator. Storage: protected from light.

5.4. Residual solvents

5.4. Residual solvents ............................................................. 5553

EUROPEAN PHARMACOPOEIA 9.5 5.4. Residual solvents
07/2018:50400 IMPURITIES: GUIDELINES

POR RESIDUAL SOLVENTS
(CHMP /ICH/82260/2006)
l. INTRODUCTION
5.4. RESIDUAL SOLVENTS
2. SCOPE OF THE GUIDELINE
LIMITING RESIDUAL SOLVENT LEVELS 3. GENERAL PRINCIPLES

IN ACTIVE SUBSTANCES, EXCIPIENTS 3.1. CLASSIFICATION OP RESIDUAL SOLVENTS BY RISK
ASSESSMENT
AND MEDICINAL PRODUCTS
3.2. METHODS POR ESTABLISHING EXPOSURE LIMITS
The International Council for Harmonisation of Technical 3.3. OPTIONS POR DESCRIBING LIMITS OP CLASS 2
Requirements for Pharmaceuticals for Human Use (ICH) has SOLVENTS
adopted Impurities Guidelines for Residual Solvents which
prescribes limits for the content of solvents which may remain 3.4. ANALYTICAL PROCEDURES
in active substances, excipients and medicinal products after 3.5. REPORTING LEVELS OP RESIDUAL SOLVENTS
processing. This guideline, the text of which is reproduced
below, excludes existing marketed products. The European 4. LIMITS OF RESIDUAL SOLVENTS
Pharmacopoeia is, however, applying the same principles 4.1. SOLVENTS TO BE AVOIDED
enshrined in the guideline to existing active substances, 4.2. SOLVENTS TO BE LIMITED
excipients and medicinal products whether or not they are the
subject of a monograph of the Pharmacopoeia. All substances 4.3. SOLVENTS WITH LOW TOXIC POTENTIAL
and products are to be tested for the content of solvents likely 4.4. SOLVENTS POR WHICH NO ADEQUATE
to be present in a substance or product. TOXICOLOGICAL DATA WAS POUND
Where the limits to be applied comply with those given below, GLOSSARY
tests for residual solvents are not generally mentioned in
specific monographs since the solvents employed may vary APPENDIX l. LIST OF SOLVENTS INCLUDED IN THE
from one manufacturer to another and the requirements of GUIDELINE
this general chapter are applied via the general monograph
on Substances for Pharmaceutical Use (2034). The competent APPENDIX 2. ADDITIONAL BACKGROUND
authority is to be informed of the solvents employed during A2.1. ENVIRONMENTAL REGULATION OP ORGANIC
the production process. This information is also given VOLATILE SOLVENTS
in the dossier submitted for a certificate of suitability of A2.2. RESIDUAL SOLVENTS IN PHARMACEUTICALS
the monographs of the European Pharmacopoeia and is
mentioned on the certificate. APPENDIX 3. METHODS FOR ESTABLISHING EXPOSURE
Where only Class 3 solvents are used, a test for loss on drying LIMITS
may be applied ora specific determination of the solvent may l. INTRODUCTION
be made. If for a Class 3 solvent a justified and authorised limit
higher than 0.5 per cent is applied, a specific determination of The objective of this guideline is to recommend acceptable
the solvent is required. amounts of residual solvents in pharmaceuticals for the safety
of the patient. The guideline recommends the use of less toxic
When Class 1 residual solvents or Class 2 residual solvents solvents and describes levels considered to be toxicologically
(or Class 3 residual solvents which exceed the 0.5 per cent) acceptable for sorne residual solvents.
are used, the methodology described in the general method
Residual solvents in pharmaceuticals are defined here as
(2.4.24) is to be applied wherever possible. Otherwise an
organic volatile chemicals that are used or produced in the
appropriate validated method is to be employed.
manufacture of active substances or excipients, or in the
When a quantitative determination of a residual solvent is preparation of medicinal products. The solvents are not
carried out, the result is taken into account for the calculation completely removed by practica} manufacturing techniques.
of the content of the substance except where a test for drying Appropriate selection of the solvent for the synthesis of active
is carried out. substance may enhance the yield, or determine characteristics
such as crystal form, purity, and solubility. Therefore, the
solvent may sometimes be a critica} parameter in the synthetic
process. This guideline <loes not address solvents deliberately
used as excipients nor <loes it address solvates. However, the
content of solvents in such products should be evaluated and
justified.
Since there is no therapeutic benefit from residual solvents, all
residual solvents should be removed to the extent possible to
meet product specifications, good manufacturing practices,
or other quality-based requirements. Medicinal products
should contain no higher levels of residual solvents than can
be supported by safety data. Sorne solvents that are known
to cause unacceptable toxicities (Class l, Table 1) should be
avoided in the production of active substances, excipients, or
medicinal products unless their use can be strongly justified
in a risk-benefit assessment. Sorne solvents associated with
less severe toxicity (Class 2, Table 2) should be limited in
order to protect patients from potential adverse effects.
Ideally, less toxic solvents (Class 3, Table 3) should be used
where practical. The complete list of solvents included in this
guideline is given in Appendix 1.
5.4. Residual solvents EUROPEAN PHARMACOPOEIA 9.5
The lists are not exhaustive and other solvents can be used Class 3 solvents: solvents with low toxic potential
and later added to the lists. Recommended limits of Class 1 Solvents with low toxic potential to man; no health-based
and 2 solvents or classification of solvents may change as new exposure limit is needed. Class 3 solvents have PDEs of 50 mg
safety data becomes available. Supporting safety data in a or more per <lay.
marketing application for a new medicinal product containing
a new solvent may be based on concepts in this guideline 3.2. METHODS POR ESTABLISHING EXPOSURE LIMITS
or the concept of qualification of impurities as expressed in The method used to establish permitted daily exposures for
the guideline for active substances (Q3A, Impurities in New residual solvents is presented in Appendix 3. Summaries of the
Active Substances) or medicinal products (Q3B, Impurities in toxicity data that were used to establish limits are published in
New Medicinal Products), or all three guidelines. Pharmeuropa, Vol. 9, No. l, Supplement April 1997.
3.3. OPTIONS POR DESCRIBING LIMITS OF CLASS 2
2. SCOPE OF THE GUIDELINE SOLVENTS
Residual solvents in active substances, excipients, and in Two options are available when setting limits for Class 2
medicinal products are within the scope of this guideline. solvents.
Therefore, testing should be performed for residual solvents Option 1 : the concentration limits in parts per million stated
when production or purification processes are known to result in Table 2 can be used. They were calculated using equation ( 1)
in the presence of such solvents. lt is only necessary to test below by assuming a product mass of 1O g administered daily.
for solvents that are used or produced in the manufacture
or purification of active substances, excipients, or medicinal . 1000 x PDE (1)
Concentrat1on (ppm) = d
product. Although manufacturers may choose to test the ose
medicinal product, a cumulative method may be used to
calculate the residual solvent levels in the medicinal product Here, PDE is given in terms of mg/day and <lose is given
from the levels in the ingredients used to produce the in g/day.
medicinal product. If the calculation results in a level equal to
or below that recommended in this guideline, no testing of the These limits are considered acceptable for all substances,
medicinal product for residual solvents need be considered. If excipients, or products. Therefore this option may be applied
however, the calculated level is above the recommended level, if the daily <lose is not known or fixed. If all excipients and
the medicinal product should be tested to ascertain whether active substances in a formulation meet the limits given
the formulation process has reduced the relevant solvent level in Option l, then these components may be used in any
to within the acceptable amount. Medicinal product should proportion. No further calculation is necessary provided
also be tested if a solvent is used during its manufacture. the daily <lose do es not exceed 1O g. Products that are
administered in doses greater than 1O g per <lay should be
This guideline <loes not apply to potential new active considered under Option 2.
substances, excipients, or medicinal products used during the
clinical research stages of development, nor <loes it apply to Option 2: it is not considered necessary for each component
existing marketed medicinal products. of the medicinal product to comply with the limits given
in Option l. The PDE in terms of mg/day as stated in
The guideline applies to all dosage forms and mutes of Table 2 can be used with the known maximum daily <lose
administration. Higher levels of residual solvents may be and equation ( 1) above to determine the concentration
acceptable in certain cases such as short term (30 days or less) of residual solvent allowed in a medicinal product. Such
or topical application. Justification for these levels should be limits are considered acceptable provided that is has been
made on a case by case basis. demonstrated that the residual solvent has been reduced to the
See Appendix 2 for additional background information related practica! mínimum. The limits should be realistic in relation
to residual solvents. to analytical precision, manufacturing capability, reasonable
variation in the manufacturing process, and the limits should
reflect contemporary manufacturing standards.
3. GENERAL PRINCIPLES
Option 2 may be applied by adding the amounts of a residual
3.1. CLASSIPICATION OP RESIDUAL SOLVENTS BY RISK solvent present in each of the components of the medicinal
ASSESSMENT product. The sum of the amounts of solvent per <lay should be
The term "tolerable daily intake" (TDI) is used by the less than that given by the PDE.
International Program on Chemical Safety (IPCS) to describe
exposure limits of toxic chemicals and "acceptable daily intake" Consider an example of the use of Option 1and Option 2
(ADI) is used by the World Health Organization (WHO) applied to acetonitrile in a medicinal product. The permitted
and other national and international health authorities and daily exposure to acetonitrile is 4.1 mg per day; thus, the
institutes. The new term "permitted daily exposure" (PDE) Option l limit is 410 ppm. The maximum administered
is defined in the present guideline as a pharmaceutically daily mass of a medicinal product is 5.0 g, and the medicinal
acceptable intake of residual solvents to avoid confusion of product contains two excipients. The composition of the
differing values for ADI's of the same substance. medicinal product and the calculated maximum content of
residual acetonitrile are given in the following table.
Residual solvents assessed in this guideline are listed in
Appendix 1 by common names and structures. They were Component Amount in Acetonitrile Daily
formulation content exposure
evaluated for their possible risk to human health and placed
into one of three classes as follows: Active substance 0.3 g 800 ppm 0.24 mg
Class 1 solvents: solvents to be avoided Excipient 1 0.9 g 400 ppm 0.36 mg
Known human carcinogens, strongly suspected human Excipient 2 3.8 g 800 ppm 3.04 mg
carcinogens, and environmental hazards. Medicinal product 5.0 g 728 ppm 3.64 mg
Class 2 solvents: solvents to be limited
Excipient 1 meets the Option l limit, but the active substance,
Non-genotoxic animal carcinogens or possible causative
excipient 2, and medicinal product do not meet the Option 1
agents of other irreversible toxicity such as neurotoxicity or
limit. Nevertheless, the product meets the Option 2 limit of
teratogenicity. 4.1 mg per day and thus conforms to the recommendations
Solvents suspected of other significant but reversible toxicities. in this guideline.

Consider another example using acetonitrile as residual 4. LIMITS OF RESIDUAL SOLVENTS

solvent. The maximum administered daily mass of a 4.1. SOLVENTS TO BE AVOIDED
medicinal product is 5.0 g, and the medicinal product contains
Solvents in Class 1 should not be employed in the
two excipients. The composition of the medicinal product
manufacture of active substances, excipients, and medicinal
and the calculated maximum content of residual acetonitrile
products because of their unacceptable toxicity or their
is given in the following table.
deleterious environmental effect. However, if their use is
Component Amount in Acetonitrile Daily unavoidable in order to produce a medicinal product with
formulation content exposure a significant therapeutic advance, then their levels should
Active substance 0.3 g 800 ppm 0.24 mg be restricted as shown in Table l, unless otherwise justified.
1, 1, 1-Trichloroethane is included in Table 1 because it is an
Excipient 1 0.9 g 2000 ppm 1.80 mg environmental hazard. The stated limit of 1500 ppm is based
Excipient 2 3.8 g 800 ppm 3.04 mg on a review of the safety data.
Medicinal product 5.0 g 1016 ppm 5.08 mg Table 1. - Class 1 solvents in pharmaceutical products
(solvents that should be avoided)
In this example, the product meets neither the Option 1 Solvent Concentration limit Concern
nor the Option 2 limit according to this summation. The m)
manufacturer could test the medicinal product to determine Benzene 2 Carcinogen
if the formulation process reduced the level of acetonitrile. If
Carbon tetrachloride 4 Toxic and environmental hazard
the level of acetonitrile was not reduced during formulation
to the allowed limit, then the manufacturer of the medicinal 1,2-Dichloroethane Toxi e
product should take other steps to reduce the amount of
1, 1-Dichloroethene 8 Toxic
acetonitrile in the medicinal product. If all of these steps fail
to reduce the level of residual solvent, in exceptional cases 1, 1, 1-Trichloroethane 1500 Environmental hazard
the manufacturer could provide a summary of efforts made
to reduce the solvent level to meet the guideline value, and 4.2. SOLVENTS TO BE LIMITED
provide a risk-benefit analysis to support allowing the product Solvents in Table 2 should be limited in pharmaceutical
to be utilised containing residual solvent at a higher level. products because of their inherent toxicity. PDEs are given to
3.4. ANALYTICAL PROCEDURES the nearest 0.1 mg/day, and concentrations are given to the
Residual solvents are typically determined using nearest 1O ppm. The stated values do not reflect the necessary
chromatographic techniques such as gas chromatography. analytical precision of determination. Precision should be
Any harmonised procedures for determining levels of residual determined as part of the validation of the method.
solvents as described in the pharmacopoeias should be used,
if feasible. Otherwise, manufacturers would be free to select Table 2. - Class 2 solvents in pharmaceutical products
the most appropriate validated analytical procedure for a Solvent PDE Concentration limit
particular application. If only Class 3 solvents are present, a (mg/day) (ppm)
non-specific method such as loss on drying may be used. Acetonitrile 4.1 410
Validation of methods for residual solvents should conform to Chlorobenzene 3.6 360
ICH guidelines "Text on Validation of Analytical Procedures" Chloroform 0.6 60
and "Extension of the ICH Text on Validation of Analytical
Procedures". Cumene 0.7 70
3.5. REPORTING LEVELS OF RESIDUAL SOLVENTS Cyclohexane 38.8 3880

Manufacturers of pharmaceutical products need certain 1,2-Dichloroethene 18.7 1870
information about the content of residual solvents in excipients
or active substances in order to meet the criteria of this Dichloromethane 6.0 600
guideline. The following statements are given as acceptable 1,2-Dimethoxyethane 1.0 100
examples of the information that could be provided from a
supplier of excipients or active substances to a pharmaceutical N,N- Dimethylacetamide 10.9 1090
manufacturer. The supplier might choose one of the following N,N-Dimethylformamide 8.8 880
as appropriate:
1,4-Dioxane 3.8 380
- only Class 3 solvents are likely to be present. Loss on drying
is less than 0.5 per cent; 2-Ethoxyethanol 1.6 160
- only Class 2 solvents X, Y, ... are likely to be present. Ali Ethyleneglycol 6.2 620
are below the Option 1 limit; Formamide 2.2 220
(Here the supplier would name the Class 2 solvents Hexane 2.9 290
represented by X, Y, ... )
Methanol 30.0 3000
- only Class 2 solvents X, Y, .. . and Class 3 solvents are
likely to be present. Residual Class 2 solvents are below 2-Methoxyethanol 0.5 50
the Option 1 limit and residual Class 3 solvents are below Methylbutylketone 0.5 50
0.5 per cent.
Methylcyclohexane 11.8 1180
If Class 1 solvents are likely to be present, they should be
identified and quantified. "Likely to be present" refers to the Methylisobutylketone 45.0 4500
solvent used in the final manufacturing step and to solvents N-Methylpyrrolidone 5.3 530
that are used in earlier manufacturing steps and not removed
consistently by a validated process. Nitromethane 0.5 50
If solvents of Class 2 or Class 3 are present at greater than Pyridine 2.0 200
their Option 1 limits or 0.5 per cent, respectively, they should Sulfolane 1.6 160
be identified and quantified.
Solvent PDE Concentration limit 4.4. SOLVENTS POR WHICH NO ADEQUA TE

(mg/day) (ppm) TOXICOLOGICAL DATA WAS FOUND
Tetrahydrofuran 7.2 720
The following solvents (Table 4) may also be of interest to
Tetralin 1.0 100 manufacturers of excipients, active substances, or medicinal
products. However, no adequate toxicological data on
Toluene 8.9 890
which to base a PDE was found. Manufacturers should
l, 1,2-Trichloroethene 0.8 80 supply justification for residual levels of these solvents in
pharmaceutical products.
Xylene* 21.7 2170
Table 4. - Solvents for which no adequate toxicological data
*usually 60 per cent m-xylene, 14 per cent p-xylene, 9 per cent o-xylene
with 17 per cent ethyl benzene.
wasfound
l, 1-Diethoxypropane Methylisopropylketone
4.3. SOLVENTS WITH LOW TOXIC POTENTIAL
l, 1-Dimethoxymethane Methyltetrahydrofuran
Solvents in Class 3 (shown in Table 3) may be regarded as less
toxic and of lower risk to human health. Class 3 includes no 2,2-Dimethoxypropane Petroleum ether
solvent known as a human health hazard at levels normally Isooctane Trichloroacetic acid
accepted in pharmaceuticals. However, there are no long-term
toxicity or carcinogenicity studies for many of the solvents in Isopropyl ether Trifluoroacetic acid
Class 3. Available data indicate that they are less toxic in acute
or short-term studies and negative in genotoxicity studies. It GLOSSARY
is considered that amounts of these residual solvents of 50 mg
per <lay or less (corresponding to 5000 ppm or 0.5 per cent Genotoxic carcinogens: carcinogens which produce cancer by
under Option 1) would be acceptable without justification. affecting genes or chromosomes.
Higher amounts may also be acceptable provided they are LOEL: abbreviation for lowest-observed effect level.
realistic in relation to manufacturing capability and good Lowest-observed effect level: the lowest <lose of substance in a
manufacturing practice. study or group of studies that produces biologically significant
increases in frequency or severity of any effects in the exposed
Table 3. - Class 3 solvents which should be limited by GMP or humans or animals.
other quality-based requirements
Modifying factor: a factor determined by professional
Acetic acid Heptane
judgement of a toxicologist and applied to bioassay data to
Aceto ne Isobutyl acetate relate that data safely to humans.
Aniso le Isopropyl acetate Neurotoxicity: the ability of a substance to cause adverse
effects on the nervous system.
1-Butanol Methyl acetate
NOEL: abbreviation for no-observed-effect leve l.
2-Butanol 3-Methyl-1-butanol No-observed-effect level: the highest <lose of substance at which
Butyl acetate Methylethylketone
there are no biologically significant increases in frequency or
severity of any effects in the exposed humans or animals.
tert-Butylmethyl ether 2-Methyl-1-propanol PDE: abbreviation for permitted daily exposure.
Dimethyl sulfoxide Pentane Permitted daily exposure: the maximum acceptable intake per
<lay of residual solvent in pharmaceutical products.
Ethanol 1-Pentanol
Reversible toxicity: the occurrence of harmful effects that are
Ethyl acetate 1-Propanol caused by a substance and which disappear after exposure to
Ethyl ether 2-Propanol the substance ends.
Strongly suspected human carcinogen: a substance for which
Ethyl formate Propyl acetate
there is no epidemiological evidence of carcinogenesis but
Formic acid Triethylamine there are positive genotoxicity data and clear evidence of
carcinogenesis in rodents.
Teratogenicity: the occurrence of structural malformations in
a developing foetus when a substance is administered during
pregnancy.
APPENDIX l. LIST OF SOLVENTS INCLUDED IN THE GUIDELINE
Solvent Other Names Structure Class
Acetic acid Ethanoic acid CH 3 COOH Class 3
Acetone 2-Propanone CH 3COCH 3 Class 3

Propan-2-one
Acetonitrile CH 3CN Class 2
Aniso le Methoxybenzene
UOCH, Class 3
o
Benzene Benzol Class 1
1-Butanol n-Butyl alcohol CH3 [CH 2] 30H Class 3

Butan-1-ol
2-Butanol sec- Butyl alcohol CH3CH 2CH(OH)CH 3 Class 3
Butan-2-ol
Butyl acetate Acetic acid butyl ester CH 3 COO[CH2 ] 3CH 3 Class 3

tert-Butylmethyl ether 2-Methoxy-2-methylpropane (CH 3 ) 3COCH 3 Class 3
Carbon tetrachloride Tetrachloromethane CCl4 Class 1
Chlorobenzene
UCI ¿::;
Class 2
Chloroform Trichloromethane CHC1 3 Class 2
Class 2
cf'cH,
Cumene Isopropylbenzene CH 3
( 1-Methylethyl)benzene
o
Cyclohexane Hexamethylene Class 2
1,2-Dichloroethane sym-Dichloroethane CH 2 ClCH 2 Cl Class 1

Ethylene dichloride
Ethylene chloride
1,1-Dichloroethene 1, 1-Dichloroethylene H 2 C=CCl2 Class 1
Vinylidene chloride
1,2-Dichloroethene 1,2-Dichloroethylene ClHC=CHCl Class 2
Acetylene dichloride
Dichloromethane Methylene chloride CH2 Cl2 Class 2
1,2-Dimethoxyethane Ethyleneglycol dimethyl ether H 3COCH 2 CH 20CH3 Class 2

Monoglyme
Dimethyl cellosolve
N,N- Dimethylacetamide DMA CH 3CON(CH3 ) 2 Class 2
N,N-Dimethylformamide DMF HCON(CH 3 ) 2 Class 2
Dimethyl sulfoxide Methylsulfinylmethane (CH 3 ) 2SO Class 3

Methyl sulfoxide
DMSO
1,4-Dioxane p-Dioxane Class 2
[l,4]Dioxane
(°) o
Ethanol Ethyl alcohol CH 3CHpH Class 3
2-Ethoxyethanol Cellosolve CH 3 CH 20CH 2 CH 2 0H Class 2
Ethyl acetate Acetic acid ethyl ester CH 3COOCH 2CH 3 Class 3
Ethyleneglycol 1,2-Dihydroxyethane HOCH2 CH 2 0H Class 2

1,2-Ethanediol
Ethyl ether Diethyl ether CH 3CH 2 0CH 2CH 3 Class 3
Ethoxyethane
l, l '-Oxybisethane
Ethyl formate Formic acid ethyl ester HCOOCH 2 CH 3 Class 3
Formamide Methanamide HCONH 2 Class 2
Formic acid HCOOH Class 3
Heptane n-Heptane CH 3 [CH2 ) 5 CH 3 Class 3
Hexane n-Hexane CH 3 (CH 2 ] 4CH 3 Class 2
Isobutyl acetate Acetic acid isobutyl ester CH3 COOCH 2 CH(CH 3 ) 2 Class 3
Isopropyl acetate Acetic acid isopropyl ester CH 3COOCH(CH3 ) 2 Class 3
Methanol Methyl alcohol CH 3 0H Class 2
2-Methoxyethanol Methyl cellosolve CHpCH 2 CH 2 0H Class 2
Methyl acetate Acetic acid methyl ester CH 3COOCH 3 Class 3
3-Methyl-1-butanol Isoamyl alcohol (CH3) 2 CHCH 2 CHpH Class 3

Isopentyl alcohol
3-Methylbutan-1-ol
Methylbutylketone 2-Hexanone CH 3 (CH2 bCOCH 3 Class 2
Hexan-2-one

Methylcyclohexane Cyclohexylmethane Class 2
Methylethylketone 2-Butanone Class 3

MEK
Butan-2-one
Methylisobutylketone 4-Methylpentan-2-one Class 2
4-Methyl-2-pentanone
MIBK
2-Methyl-1-propanol Isobutyl alcohol Class 3
2-Methylpropan-1-ol
N- Methylpyrrolidone 1-Methylpyrrolidin-2-one Class 2
l -Methyl-2-pyrrolidinone
Nitromethane CH 3 N0 2 Class 2
Pentane n-Pentane CH 3 [CH2 ] 3 CH 3 Class 3
1-Pentanol Amyl alcohol CH 3 [CH 2LCHpH Class 3
Pentan-1-ol
Pentyl alcohol
1-Propanol Propan-1-ol Class 3
Propyl alcohol
2-Propanol Propan-2-ol Class 3
Isopropyl alcohol
Propyl acetate Acetic acid propyl ester Class 3
Pyridine Class 2
o
N
Suifonane Tetrahydrothiophene 1, 1-dioxide Class 2

o o
Tetrahydrofuran Tetramethylene oxide

ó Class 2
Oxacyclopentane
ó
co
Tetralin 1,2,3,4-Tetrahydronaphthalene Class 2
Toluene Methylbenzene
UCH, Class 2
1,1,1-Trichloroethane Methylchloroform CH3 CCl3 Class 1
1,1,2-Trichloroethene Trichloroethene HC1C=CC1 2 Class 2

Triethylamine N,N-Diethylethanamine N(CH 2 CH 3 ) 3 Class 3
Xylene* Dimethybenzene Class 2
Xylol
*usually 60 per cent m-xylene, 14 per cent p-xylene, 9 per cent o-xylene with 17 per cent ethyl benzene.
APPENDIX 2. ADDITIONAL BACKGROUND health and maintenance of environmental integrity against

A2.1. ENVIRONMENTAL REGULATION OF ORGANIC the possible deleterious effects of chemicals resulting from
VOLATILE SOLVENTS long-term environmental exposure. The methods involved in
Severa! of the residual solvents frequently used in the the estimation of maximum safe exposure limits are usually
production of pharmaceuticals are listed as toxic chemicals based on long-term studies. When long-term study data
in Environmental Health Criteria (EHC) monographs and are unavailable, shorter term study data can be used with
the Integrated Risk Information System (IRIS). The objectives modification of the approach such as use of larger safety
of such groups as the International Programme on Chemical factors. The approach described therein relates primarily to
Safety (IPCS), the United States Environmental Protection long-term or life-time exposure of the general population in
Agency (USEPA) and the United States Food and Drug the ambient environment, i.e. ambient air, food, drinking
Administration (USFDA) include the determination of water and other media.
acceptable exposure levels. The goal is protection of human

A2.2. RESIDUAL SOLVENTS IN PHARMACEUTICALS "safety factors" in Pharmacopoeial Forum. The assumption
Exposure limits in this guideline are established by referring of 100 per cent systemic exposure is used in all calculations
to methodologies and toxicity data described in EHC and regardless of route of administration.
IRIS monographs. However, sorne specific assumptions about The modifying factors are as follows:
residual solvents to be used in the synthesis and formulation
of pharmaceutical products should be taken in to account in F1 = a factor to account for extrapolation between species:
establishing exposure limits. They are:
F1 2 for extrapolation from dogs to humans;
1) Patients (not the general population) use pharmaceuticals
to treat their diseases or for prophylaxis to prevent infection Fl 2.5 for extrapolation from rabbits to humans;
or disease.
2) The assumption of life-time patient exposure is not Fl 3 for extrapolation from monkeys to humans;
necessary for most pharmaceutical products but may be Fl 5 for extrapolation from rats to humans;
appropriate as a working hypothesis to reduce risk to human
health. Fl 1O for extrapolation from other animals to
3) Residual solvents are unavoidable components in humans;
pharmaceutical production and will often be a part of Fl 12 for extrapolation from mice to humans.
medicinal products.
4) Residual solvents should not exceed recommended levels F 1 takes into account the comparative surface area: body
except in exceptional circumstances. weight ratios for the species concerned and for man. Surface
are a (S) is calculated as :
5) Data from toxicological studies that are used to determine
acceptable levels for residual solvents should have been S = kmo.67
generated using appropriate protocols such as those described
for example, by OECD, EPA, and the FDA Red Book. in which m = body mass, and the constant k has been taken to
be 10. The body weight used in the equation are those shown
APPENDIX 3. METHODS FOR ESTABLISHING EXPOSURE below in Table A3.-l.
LIMITS
Table A3.-l. - Values used in the calculations in this document
The Gaylor-Kodell method of risk assessment (Gaylor, D. W.
and Kodell, R. L. Linear Interpolation algorithm for low <lose Rat body weight 425 g
assessment of toxic substance. J. Enviran. Pathology, 4, 305, Pregnant rat body weight 330 g
1980) is appropriate for Class 1 carcinogenic solvents. Only in
cases where reliable carcinogenicity data are available should Mouse body weight 28 g
extrapolation by the use of mathematical models be applied to Pregnant mouse body weight 30 g
setting exposure limits. Exposure limits for Class 1 solvents
could be determined with the use of a large safety factor (i.e., Guinea-pig body weight 500 g
10 000 to 100 000) with respect to the no-observed-effect level Rhesus monkey body weight 2.5 kg
(NOEL). Detection and quantification of these solvents should
be by state-of-the-art analytical techniques. Rabbit body weight (pregnant or not) 4 kg
Acceptable exposure levels in this guideline for Class 2 Beagle dog body weight 11.5 kg
solvents were established by calculation of PDE values
Rat respiratory volume 290 L/day
according to the procedures for setting exposure limits in
pharmaceuticals (Pharmacopeial Forum, Nov-Dec 1989), Mouse respiratory volume 43 L/day
and the method adopted by IPCS for Assessing Human
Rabbit respiratory volume 1440 L/day
Health Risk of Chemicals (Environmental Health Criteria
170, WHO, 1994). These methods are similar to those used Guinea-pig respiratory volume 430 L/day
by the USEPA (IRIS) and the USFDA (Red Book) and others.
Human respiratory volume 28800 L/day
The method is outlined here to give a better understanding of
the origin of the PDE values. It is not necessary to perform Dog respiratory volume 9000 L/day
these calculations in order to use the PDE values tabulated in
Monkey respiratory volume ll50 L/day
Section 4 of this document.
PDE is derived from the no-observed-effect level (NOEL), or Mouse water consumption 5 mL/day
the lowest-observed effect level (LOEL), in the most relevant Rat water consumption 30 mL/day
animal study as follows:
Rat food consumption 30 g/day
PDE = NOEL x Weight Adjustment
Fl X F2 X F3 X F4 X F5 F2 = a factor of 1O to account for variability between
The PDE is derived preferably from a NOEL. If no NOEL individuals.
is obtained, the LOEL may be used. Modifying factors A factor of 1O is generally given for all organic
proposed here, for relating the data to humans, are the same solvents, and 10 is used consistently in this guideline.
kind of "uncertainty factors" used in Environmental Health
Criteria (Environmental Health Criteria 170, World Health F3 a variable factor to account for toxicity studies of
Organization, Geneva, 1994), and "modifying factors" or short-term exposure:
F3 1 for studies that last at least one half-lifetime calculation. It is recognised that sorne adult patients weigh less
(1 year for rodents or rabbits; 7 years for cats, than 50 kg; these patients are considered to be accommodated
dogs and monkeys); by the built-in safety factors used to determine a PDE. If the
solvent was present in a formulation specifically intended for
F3 1 for reproductive studies in which the whole paediatric use, an adjustment for a lower body weight would
period of organogenesis is covered; be appropriate.
F3 As an example of the application of this equation, consider the
2 for a 6 month study in rodents, or a 3.5 year
toxicity study of acetonitrile in mice that is summarised in
study in non-rodents;
Pharmeuropa, Vol. 9. No. 1, Supplement, April 1997, page S24.
F3 5 for a 3 month study in rodents, or a 2 year The NOEL is calculated to be 50.7 mg kg- 1 day-1• The PDE for
study in non-rodents; acetonitrile in this study is calculated as follows:
F3 10 for studies of a shorter duration. 1 1

PDE = 50.7mg kg- day- x 50 kg = 4.22 m da -1
In all cases, the higher factor has been used for study durations 12 X 10 X 5 X 1 X 1 g y
between the time points, e.g. a factor of 2 for a 9 month
rodent study. In this example,
F4 = a factor that may be applied in cases of severe toxicity, Fl 12 to account for the extrapolation from mice to
e.g. non-genotoxic carcinogenicity, neurotoxicity or humans;
teratogenicity.
F2 10 to account for differences between individual
In studies of reproductive toxicity, the following humans;
factors are used:
F3 5 because the duration of the study was only
F4 1 for foetal toxicity associated with maternal 13 weeks;
toxicity;
F4 1 because no severe toxicity was encountered;
F4 5 for foetal toxicity without maternal toxicity;
F5 1 because the no-effect level was determined.
F4 5 for a teratogenic effect with maternal toxicity;
The equation for an ideal gas, PV = nRT, is used to convert
F4 1O for a teratogenic effect without maternal concentrations of gases used in inhalation studies from units
toxicity. of ppm to units of mg/L or mg/m 3 • Consider as an example
the rat reproductive toxicity study by inhalation of carbon
F5 = a variable factor that may be applied if the no-effect tetrachloride (molecular weight 153.84) summarised in
level was not established. Pharmeuropa, Vol. 9, No. l, Supplement, April 1997, page S9.
When only a LOEL is available, a factor of up to 10 can be n p 300 x 10- 6 atm x 153 840 mg mol- 1
used depending on the severity of the toxicity. V RT 0.082 L atm K- 1 mo1- 1 x 298 K
The weight adjustment assumes an arbitrary adult human
body weight for either sex of 50 kg. This relatively low weight = 46.15 mg = 1. 89 m /L
24.45 L g
provides an additional safety factor against the standard
weights of 60 kg or 70 kg that are often used in this type of The relationship 1000 L = 1 m 3 is used to convert to mg/m 3•

5.12. Reference standards

5.12. Reference standards ...................................................... 5563

EUROPEAN PHARMACOPOEIA 9.5 5.12. Reference standards
07/2018:51200 Reference material (RM). A material sufficiently homogeneous

and stable with respect to one or more specified properties,
which has been established to be fit for its intended use in the
measurement process.
Certified reference material (CRM). A reference material
5.12. REFERENCE STANDARDS characterised by a metrologically valid procedure for one
or more specified properties, accompanied by a certificate
that states the value of the specified property, its associated
This chapter is published for information. uncertainty, and a statement of metrological traceability.
l. INTRODUCTION
3. USE OF EUROPEAN PHARMACOPOEIA REFERENCE
'Reference standard' is used in this chapter as a general term STANDARDS
covering reference substances, reference preparations and
reference spectra. European Pharmacopoeia reference standards are employed
in the identification, purity testing and assay of articles subject
Reference standards are frequently necessary to achieve to a European Pharmacopoeia monograph or general chapter.
adequate quality control of medicinal products and their European Pharmacopoeia reference standards are shown to be
components. suitable for their intended purpose; they are not necessarily
Reference standards are established using suitable procedures suitable for other purposes. If a European Pharmacopoeia
and their continued suitability for use is monitored according reference standard is to be used for any purpose other than
to a predefined programme. Where a reference standard that for which it has been established, its suitability for the
is needed, it is an integral part of the pharmacopoeial new use has to be fully demonstrated and when applicable, to
monograph or the manufacturer's specification. Where a be described in the marketing authorisation application. Any
European Pharmacopoeia reference standard is referred to value assigned to a reference standard is valid for the intended
in a monograph or general chapter, it represents the official use and not necessarily for other uses.
standard that is alone authoritative in case of doubt or dispute.A European Pharmacopoeia reference standard with an
2. TERMINOLOGY assigned content/potency for use in the assay of a substance
for pharmaceutical use (see general monograph Substances
Primary standard. A standard designated or widely for pharmaceutical use (2034)) may be suitable to determine
acknowledged as having the highest metrological qualities the content/potency of that substance in a pharmaceutical
and whose property value is accepted without reference to preparation provided all of the following conditions are
other standards of the same property or quantity, within a fulfilled:
specified context. This definition does not cover international
standards. - the chromatographic assay method described in the active
substance monograph is employed;
International standard. A primary standard provided to
enable the results of biological or immunological assay - the applicability of the method to the particular
procedures to be expressed in the same way throughout the pharmaceutical preparation (absence of interference) is
world. The value assignment is in International Units (IU) or verified by the user;
another suitable unit. The unitage is normally assigned to a - any pre-treatment of the sample (e.g. extraction, filtration)
first international standard by the World Health Organization is validated for the particular pharmaceutical preparation.
(WHO) in an arbitrary manner based on an international
It is the policy of the European Pharmacopoeia to supply
interlaboratory study. Activities in International Units are
reference standards in adequate quantities for immediate use
assigned to replacement international standards, where
(i.e. the quantity needed to perform the test(s) described in the
appropriate, by comparing them with a previous standard.
monograph or general chapter) after opening of the container.
Secondary standard. A standard whose property value is Use in other conditions is the responsibility of the analyst.
assigned by comparison with a primary standard of the same If an unopened container is stored in the recommended
property or quantity. conditions, it remains suitable for use as long as it is of the
European Pharmacopoeia reference standard. A reference current batch. Information on current batch numbers is
standard established under the aegis of and adopted by the provided in the European Pharmacopoeia reference standards
European Pharmacopoeia Commission. database (http://go.edqm.eu/RS). Storage of reconstituted or
European Pharmacopoeia chemical reference substance (CRS). diluted solutions of reference standards is not recommended
A substance or mixture of substances intended for use as unless suitability has been demonstrated by the user.
stated in a monograph or general chapter of the European Secondary standards. A secondary standard is usually
Pharmacopoeia. CRSs are in general primary standards, established to reduce the use of the primary standard and may
except for those (notably antibiotics) that are calibrated be used for routine quality control purposes. A secondary
in International Units. The latter are secondary standards standard shall exhibit the same property or properties as the
traceable to the international standard. primary standard to which it is traceable. It shall therefore be
European Pharmacopoeia herbal reference standard (HRS). A used for the same purpose as the primary standard.
herbal drug preparation (usually an extract) or a herbal drug International standards are usually available in relatively
intended for use as stated in a monograph or general chapter limited quantities and are intended to be used for the
of the European Pharmacopoeia. Unless otherwise specified, characterisation and calibration of secondary standards; these
HRSs are designated as primary reference standards for their secondary standards may then be used as working standards.
intended use.
European Pharmacopoeia biological reference preparation 4. ESTABLISHMENT OF REFERENCE STANDARDS
(BRP). A substance or mixture of substances intended for use 4-1. PRIMARY STANDARDS
as stated in a monograph or general chapter of the European A substance or preparation to be established as a primary
Pharmacopoeia. BRPs are either secondary standards standard is characterised by a variety of analytical techniques
calibrated in International Units or primary standards, which chosen to demonstrate its suitability for use.
may be used to define a European Pharmacopoeia Unit
Por reference standards used for the control of medicinal
-
(Ph. Eur. U.). Other assigned contents may also be used, for
example, virus titre or number of bacteria. products and their components, relevant parts of the following
test programme are usually applied.
5.12. Reference standards EUROPEAN PHARMACOPOEIA 9.5
Test programme: If an impurity is not available in a sufficient quantity to

establish a CRS, a number of other options exist:
- Characterisation of the substance (structural elucidation) by
appropriate chemical attributes such as structural formula, - preparation of a CRS that contains a mixture of the
empirical formula, molecular mass or composition. compound(s) and the impurity or impurities;
A number of techniques may be used including: - preparation of a CRS containing a mixture of specified
- nuclear magnetic resonance spectrometry; impurities.
Where such a mixture is also used to determine the content
- mass spectrometry;
of a given impurity, the content of the impurity in the CRS is
- infrared spectrophotometry; determined by appropriate separation methods and a content
- elemental analysis. is assigned to the reference standard.
4-2-3. Assay
- Determination of purity:
4-2-3-1. Chemical assay. When a CRS is to be used for
- determination of the content of related substances by an quantitative determination of a substance for pharmaceutical
appropriate separation technique and/or spectrometric use (assay standard), the extent of testing is greater than when
method, where applicable; a CRS is used for other purposes. Unless the substance is
- quantitative determination of water; of high purity, severa! collaborating laboratories are usually
involved in testing. The results obtained are used to assign a
- determination of the content of residual solvents; content. It is particularly important to quantify the impurities
- determination of loss on drying, which may in certain if a selective assay is employed. In such a case, it is best to
circumstances replace the determinations of water and characterise the candidate substance by additional analytical
residual solvents; procedures that are scientifically justified, including, where
possible, independent methods and methods based on
- determination of inorganic impurities (e.g. sulfated ash, different principies.
atomic absorption spectrometry, inductively coupled
plasma spectrometry, X-ray fluorescence spectrometry); For a European Pharmacopoeia chemical reference substance
the results are usually not used to determine an assigned established for assay purposes, the assigned content is
content, except where they would have an appreciable usually calculated from the values obtained from the analyses
impact upon it; performed for the determination of impurities (organic,
inorganic, water and solvents) by applying the principie of
- determination of purity by an independent method mass balance; other suitable methods may also be used. When
(e.g. quantitative nuclear magnetic resonance possible, the assigned content is confirmed by comparing with
spectrometry, differential scanning calorimetry or the result obtained by an independent method.
titration where appropriate; the results of these
determinations are usually used to support and confirm If a CRS is required for a non-chromatographic assay method
the results obtained from separation techniques; they (e.g. colorimetry or ultraviolet spectrophotometry), the
are not used in the calculation of the assigned content). relative reactivity or relative absorbance of the impurities
present in a substance must be checked to ensure that they are
For biologicals, guidance is given in the WHO not markedly different from those of the substance.
recommendations for the preparation, characterisation and Unless otherwise stated, an assigned content is given for the
establishment of international and other biological reference substance or preparation as presented in the container ('as
standards (WHO Technical Report Series). is'), and the contents are not to be dried before use. For assay
4-2. EUROPEAN PHARMACOPOEIA CHEMICAL standards prepared by lyophilisation, the content of the pure
REFERENCE SUBSTANCES substance is indicated in milligrams or International Units
The extent of testing and the number of laboratories involved per vial.
in the establishment of a CRS depend on the use of the CRS 4-2-3-2. Microbiological assay. The potency is expressed in
and are tailored to ensure fitness for purpose. International Units or in European Pharmacopoeia Units if no
Where an interlaboratory study is carried out during international standard exists. The assigned potency together
establishment, a protocol is provided for each participant with the confidence limits are calculated from statistically
and only valid results derived according to the protocol are valid results of an interlaboratory study, according to the usual
used to determine an assigned content or otherwise confirm statistical procedures (5.3).
suitability. 4-2-3-3. Assay of components of herbal drugs and herbal drug
preparations. Reference standards used in monographs on
Relevant parts of the following programme are typically
applied. herbal drugs vary in the extent of testing depending on the
type of referen ce standard.
4-2-1. Identification. In general, the candidate batch is shown
An active component or marker constituent used as a CRS is
to comply with the relevant requirements of the monograph;
usually characterised and evaluated for identity and purity; a
full structural elucidation is carried out for the first batch.
value for content is assigned irrespective of the purity.
4-2-2. Related substances test. A CRS corresponding to
4-2-4. Establishment report. A report containing the results
an impurity is characterised for identity and purity. Where of the establishment study as well as information concerning
a CRS is used to determine the content of a given impurity,
the use of the CRS is prepared by EDQM, approved by the
the preferred mínimum content is 95.0 per cent; where this
relevant group of experts and adopted by the European
is achieved the assigned content of the CRS is not given and Pharmacopoeia Commission. The report for an assay standard
it is considered to be 100.0 per cent; this approximation is
indicates the content assigned to the substance with the
acceptable since there will be no appreciable effect on the
rationale for this assignment. The estimated uncertainty of
determination of impurities. When this minimum content
the assigned content is calculated, and where it is less than
cannot be obtained, an assigned content is given to the CRS.
a predefined value, which is considered to be negligible in
CRSs used to determine the content of a given impurity are relation to the acceptance criteria for the assay, then the study
normally in the same acid, base or salt form as the substance is accepted. Otherwise, the study may be repeated, in whole or
that is the subject of the corresponding monograph. Where in part, or the limits defined for the pharmaceutical substance
this is not the case, unless otherwise justified, a corresponding may be widened. The uncertainty of the assigned content is
stoichiometric conversion factor is applied. usually not given as part of the information provided with

EUROPEAN PHARMACOPOEIA 9.5 5.12. Reference standards
the CRS, since the precision of the method and the uncertainty Pharmacopoeia Commission. The establishment reports
of the content assigned to the CRS are taken into account are published in Pharmeuropa Bio & Scientific Notes
when setting the limit(s) in a monograph. (http://pharmeuropa.edqm.eu/PharmeuropaBioSN).
4-3. EUROPEAN PHARMACOPOEIA HERBAL REFERENCE 4-5. SECONDARY STANDARDS
STANDARDS A secondary standard should exhibit the same property or
A HRS is used when the available quantity of pure properties as the primary standard, relevant for the test(s)
constituent to be assayed is considered either insufficient or for which it is established. The extent of testing may not be
non-sustainable. HRSs may also be established for purposes as comprehensive as is required for the establishment of a
other than assays, notably for use in tests for adulteration or primary standard. The secondary standard is established by
for system suitability. A HRS is characterised by a variety of comparison with the primary standard to which it is traceable.
analytical techniques chosen to demonstrate its suitability An official primary standard is used wherever possible for
for the intended use; relevant parts of the following test establishment of secondary standards.
programme may be applied.
5. MANUFACTURING, LABELLING, STORAGE AND
Test programme: DISTRIBUTION OF EUROPEAN PHARMACOPOEIA
- macroscopy; REFERENCESTANDARDS
- microscopy; 5-1. MANUFACTURING
- thin-layer chromatography; All operations are carried out according to the relevant
best practices to ensure the traceability and integrity of the
- gas chromatography;
reference standard. The manufacturing record includes
- liquid chromatography; information regarding filling, labelling and storage. Reference
- quantitative determination of water; standards are dispensed into containers under appropriate
filling and closure conditions to ensure the integrity of
- content of residual solvents;
the reference standard. The containers employed may be
- loss on drying; multi-use or single-use, but the latter is preferred to minimise
- foreign matter; the risk of decomposition, contamination or water uptake.
- assay of constituents (e.g. constituents with known 5-2. LABELLING
therapeutic activity, active markers, analytical markers) The label bears the name of the reference standard, the name
relevant to the intended use of the reference standard. and address of the supplier, the batch number and unit
The extent of testing and the number of laboratories involved quantity (quantity per vial/ampoule).
in the establishment of a HRS depend on its intended use. An accompanying leaflet, considered as part of the labelling,
For a European Pharmacopoeia herbal reference standard is normally provided.
used for assay purposes, the assigned content is usually If used as an assay standard, the following information is also
established by an interlaboratory study, using the assay given:
method specified in the individual monograph in which the - the assigned percentage content;
reference standard is intended to be used, comparing against - or, the content in milligrams or millilitres of the chemical
a suitable pure sample of the constituent or constituents for entity in the container;
which the -content is to be assigned.
- or, the assigned potency (for biological assays or
Establishment report. The establishment report for the HRS microbiological assays) in units either per milligram, per
is prepared in the same manner as CRSs (see section 4-2-4). millilitre or per vial/ampoule.
4-4. EUROPEAN PHARMACOPOEIA BIOLOGICAL For European Pharmacopoeia reference standards, no re-test
REFERENCE PREPARATIONS AND CHEMICAL or expiry date is given since the re-test programme (see
REFERENCE SUBSTANCES POR BIOLOGICALS section 6) monitors continued fitness for use. A batch validity
Most BRPs and sorne CRSs used for the testing of biological statement (BVS) for each European Pharmacopoeia reference
substances and preparations are established through the standard is available from the European Pharmacopoeia
Biological Standardisation Programme, under the aegis reference standards database (http://go.edqm.eu/RS).
of the Council of Europe and the European Commission. 5-3. STORAGE AND DISTRIBUTION
These reference standards are usually secondary standards
Reference standards are to be stored and distributed in
calibrated against the corresponding WHO international
standard. Where no international standard is available, they conditions suitable to ensure optimal stability.
are primary standards with an assigned content/potency in Most European Pharmacopoeia reference standards are stored
European Pharmacopoeia Units or another suitable unit. in temperature-controlled rooms at 5 ± 3 ºC. However, a
They are established through interlaboratory studies where number of reference standards are stored at - 20 ± 5 ºC and
participating laboratories test the candidate material(s) and sorne (e.g. live virus preparations) are stored at - 80 ± 10 ºC
valid data are used to assign the official potency/content. Sorne or at - 196 ºC to - 170 ºC under liquid nitrogen.
of these studies are jointly organised with other organisations Appropriate packaging is used to minimise the risk of damage
to establish a common material or batch of material as during transport, to keep the reference standard at the
standard. In these cases, although the material constituting the appropriate temperature when necessary and to comply with
European Pharmacopoeia reference standard may be identical the current transport regulations.
to the international standard and its use may be validated Reference standards that are stored at 5 ± 3 ºC are normally
through the same interlaboratory study, it is considered as a transported without cooling when short-term excursions from
secondary standard for use as a working standard. the long-term storage temperature are not deleterious to the
The study reports are endorsed by the study participants reference standard. Sorne of them may nevertheless be sent
and approved by the relevant European Pharmacopoeia at + 5 ºC, packed with cold packs in cases where an increase
group of experts, where applicable, and by the Steering in temperature is detrimental to their stability. Reference
Committee of the Biological Standardisation Programme. standards stored at - 20 ºC are packed with cold packs or
The study results are then submitted to the European on dry ice (solid carbon dioxide) and dispatched by express
Pharmacopoeia Commission. Reference standards/materials courier or air freight. Reference standards stored at - 80 ºC
established through the Biological Standardisation or stored under liquid nitrogen are packed on solid carbon
Programme are officially adopted by the European di oxide.
5.12. Reference standards EUROPEAN PHARMACOPOEIA 9.5
Delivery conditions, dispatch and storage temperatures are The periodicity and extent of re-testing reference standards
available in the Reference Standards Catalogue, the Terms of depends on a number of factors including:
Supply and the European Pharmacopoeia reference standards
database (http://go.edqm.eu/RS). - stability;
6. RE-TEST PROGRAMME OF EUROPEAN - container and closure system;

PHARMACOPOEIA STANDARDS - storage conditions;
A system is established and implemented to ensure the
continued fitness-for-use of the European Pharmacopoeia - hygroscopicity;
reference standards. Normally, a re-test programme is applied, - physical form;
taking account of the known physico-chemical properties
and stability data for the reference standard. Reference - intended use;
standards are periodically tested for stability during storage. - presentation (single use/multiple use).
A monitoring programme is applied that is designed to detect
at an early stage any sign of degradation using appropriate The re-test period may be lengthened with the support of
analytical techniques. The methods employed are typically sufficient data. The maximum permitted variation from the
chosen from amongst those performed during establishment assigned content should be pre-defined, and if exceeded, the
of the reference standard so that baseline data is available. batch should be re-established or replaced.

General monographs
Pharmaceutical preparations ................................................. 5569 Vaccines for veterinary use .................................................... 5574
Vaccines for human use ......................................................... 5571

EUROPEAN PHARMACOPOEIA 9.5 Pharmaceutical preparations
07/2018:2619 without an appropriate marketing authorisation. The

exemptions from the formal licensing requirement allow
the supply of unlicensed products to meet the special needs
of individual patients. However, when deciding to use an
unlicensed preparation all health professionals involved
PHARMACEUTICAL PREPARATIONS (e.g. the prescribing practitioners and/or the preparing
pharmacists) have, within their area of responsibilities, a duty
of careto the patient receiving the pharmaceutical preparation.
Pharmaceutica In considering the preparation of an unlicensed pharmaceutical
INTRODUCTION preparation, a suitable level of risk assessment is undertaken.
This monograph is intended to be a reference source The risk assessment identifies:
of standards in the European Pharmacopoeia on active - the criticality of different parameters (e.g. quality of
substances, excipients and dosage forms, which are to be active substances, excipients and containers; design
applied in the manufacture/preparation of pharmaceuticals, of the preparation process; extent and significance of
but not a guide on how to manufacture as there is specific testing; stability of the preparation) to the quality of the
guidance available covering methods of manufacture and preparation; and
associated controls.
- the risk that the preparation may present to a particular
It <loes not cover investigational medicinal products, but patient group.
competent authorities may refer to pharmacopoeial standards
when authorising clinical trials using investigational medicinal Based on the risk assessment, the person responsible for the
preparation must ensure, with a suitable level of assurance,
products.
that the pharmaceutical preparation is, throughout its
DEPINITION shelf-life, of an appropriate quality and suitable and fit for
Pharmaceutical preparations are medicinal products generally its purpose. Por stock preparations, storage conditions and
consisting of active substances that may be combined with shelf-life have to be justified on the basis of, for example,
excipients, formulated into a dosage form suitable for the analytical data or professional judgement, which may be based
intended use, where necessary after reconstitution, presented on literature references.
in a suitable and appropriately labelled container. PRODUCTION
Pharmaceutical preparations may be licensed by the competent
Manufacture/preparation must take place within the
authority, or unlicensed and made to the specific needs of
framework of a suitable quality system and be compliant with
patients according to legislation. There are 2 categories of
the standards relevant to the type of product being made.
unlicensed pharmaceutical preparations:
Licensed products must comply with the requirements of
- extemporaneous preparations, i.e. pharmaceutical their licence. Por unlicensed products a risk assessment as
preparations individually prepared for a specific patient or outlined in the section 'Ethical considerations and guidance
patient group, supplied after preparation; in the preparation of unlicensed pharmaceutical preparations'
- stock preparations, i.e. pharmaceutical preparations is of special importance, as these products are not previously
prepared in advance and stored until a request for a supply assessed by the competent authority.
is received. Formulation. During pharmaceutical development or prior
In addition to this monograph, pharmaceutical preparations to manufacture/preparation, suitable ingredients, processes,
also comply with the General Notices and with the relevant tests and specifications are identified and justified in order to
general chapters of the Pharmacopoeia. General chapters are ensure the suitability of the product for the intended purpose.
normally given for information and become mandatory when This includes consideration of the properties required in order
referred to in a general or specific monograph, unless such to identify whether specific ingredient properties or process
reference is made in a way that indicates that it is not the steps are critica! to the required quality of the pharmaceutical
intention to make the text referred to mandatory but rather to preparation.
cite it for information.
Active substances and excipients. Active substances
Where relevant, pharmaceutical preparations also and excipients used in the formulation of pharmaceutical
comply with the dosage form monographs (e.g. preparations comply with the requirements of the relevant
Capsules (0016), Tablets (0478)) and general monographs general monographs, e.g. Substances for pharmaceutical
relating to pharmaceutical preparations (e.g. Allergen use (2034), Essential oils (2098), Herbal drug extracts (0765),
products (1063), Herbal teas (1435), Homoeopathic Herbal drugs (1433), Herbal drug preparatíons (1434), Herbal
preparations (1038), Homoeopathic pillules, coated (2786), drugs for homoeopathic preparations (2045), Mother tinctures
Homoeopathic pillules, impregnated (2079), Immunosera for homoeopathic preparations (2029), Methods of preparation
for human use, animal (0084), Immunosera for veterinary of homoeopathic stocks and potentisation (2371), Products
use (0030), Monoclonal antibodies for human use (2031), offermentation (1468), Products with risk of transmitting
Radiopharmaceutical preparatíons (0125), Vaccines for human agents of animal spongiform encephalopathies (1483), Products
use (0153), Vaccines for veterinary use (0062)). of recombinant DNA technology (0784), Vegetable fatty
Where pharmaceutical preparations are manufactured/ oils (1579).
prepared using materials of human or animal origin, In addition, where specific monographs exist, the quality of
the general requirements of general chapters 5.1. 7. Viral the active substances and excipients used complies with the
safety, 5.2.6. Evaluation of safety of veterinary vaccines and corresponding monographs.
immunosera and 5.2.8. Minimísing the risk of transmitting
animal spongiform encephalophathy agents via human and Where no specific monographs exist, the required quality
veterinary medicinal products apply, where appropriate. must be defined, taking into account the intended use and
the involved risk.
ETHICAL CONSIDERATIONS AND GUIDANCE IN THE When physicochemical characteristics of active substances
PREPARATION OP UNLICENSED PHARMACEUTICAL and functionality-related characteristics (PRCs) of excipients
PREPARATIONS (e.g. particle-size distribution, viscosity, polymorphism) are
The underlying principie of legislation for pharmaceutical critica! in relation to their role in the manufacturing process
preparations is that, subject to specific exemptions, no and quality attributes of the pharmaceutical preparation, they
pharmaceutical preparation may be placed on the market must be identified and controlled.
Pharmaceutical preparations EUROPEAN PHARMACOPOEIA 9.5
Detailed information on FRCs is given in general chapter 5.15. The following tests are applicable to many preparations and
Functionality-related characteristics of excipients. are therefore listed here.
Microbiological quality. The formulation of the Appearance. The appearance (e.g. size, shape and colour) of
pharmaceutical preparation and its container must ensure that the pharmaceutical preparation is controlled.
the microbiological quality is suitable for the intended use. Identity and purity tests. Where applicable, the following
During developrnent, it shall be dernonstrated that the tests are carried out on the pharmaceutical preparation:
antimicrobial activity of the preparation as such or, if - identification of the active substance(s);
necessary, with the addition of a suitable preservative or - identification of specific excipient(s), such as preservatives;
preservatives, or by the selectíon of an appropriate container,
provides adequate protection from adverse effects that may - purity tests (e.g. investigation of degradation products,
arise from microbial contarnination or proliferation during residual solvents (2.4.24) or other related impurities,
the storage and use of the preparatíon. A suitable test method sterílity (2.6.1));
together with criteria for evaluating the preservative properties - safety tests (e.g. safety tests for biological products).
of the forrnulation are provided in general chapter 5.1.3. Elemental impurities. General chapter 5.20. Elemental
Efficacy of antimicrobial preservation. impurities applies to pharrnaceutical preparatíons except
If preparations do not have adequate antirnicrobial efficacy products for veterinary use, unlicensed preparations and other
and do not contain antimicrobial preservatives they are products that are excluded from the scope of this chapter.
supplied in single-dose containers, or in multidose containers For pharmaceutical preparations outside the scope of general
that prevent microbial contamination of the contents after chapter 5.20, manufacturers of these products remain
opening. responsible for controlling the levels of elemental irnpurities
In the manufacture/preparation of non-sterile pharmaceutical using the principies of risk rnanagement.
preparations, suitable measures are taken to ensure their If appropriate, testing is performed using suitable analytical
microbial quality; recornmendations on this aspect are procedures according to general chapter 2.4.20. Determination
provided in general chapters 5.1.4. Microbiological quality of elemental impurities.
of non-sterile pharmaceutical preparations and substances Uniformity (2.9.40 or 2.9.5/2.9.6). Pharmaceutical
for pharmaceutical use and 5.1.8. Microbiological quality of preparations presented in single-dose units comply with the
herbal medicinal products for oral use and extracts used in test(s) as prescribed in the relevant specific dosage form
their preparation. rnonograph. If justified and authorised, general chapter 2.9.40
Sterile preparations are rnanufactured/prepared using can be applicable only at the time of release.
materials and rnethods designed to ensure sterílity and to Special uniformity requirements apply in the following cases:
avoid the introduction of contaminants and the growth
of micro-organisms; recornrnendations on this aspect are - for herbal drugs and herbal drug preparations, cornpliance
provided in general chapter 5.1.1. Methods of preparation of with general chapter 2.9.40 is not required;
sterile products. - for homoeopathic preparations, the provisions of general
chapters 2.9.6 and 2.9.40 are normally not appropriate,
Containers. A suitable container is selected. Consideration is however in certain circurnstances compliance with these
given to the intended use of the preparation, the properties of chapters rnay be required by the competent authority;
the container, the required shelf-life, and product/container
incompatibilities. Where applicable, containers for - for single- and multivitarnin and trace-elernent
pharrnaceutical preparations comply with the requirements preparations, compliance with general chapters 2. 9. 6 and
for containers (3.2 and subsections) and rnaterials used for the 2.9.40 (content uniformity only) is not required;
manufacture of containers (3.1 and subsections). - in justified and authorised circumstances, for other
preparations, cornpliance with general chapters 2.9.6 and
Stability. Stability requirements of pharrnaceutical
2.9.40 may not be required by the competent authority.
preparations are dependent on their intended use and on the
desired storage time. Reference standards. Reference standards may be needed
at various stages for quality control of pharmaceutical
Where applicable, the probability and criticality of possible
preparations. They are established and monitored taking due
degradation products of the active substance(s) and/or
account of general chapter 5.12. Reference standards.
reaction products of the active substance(s) with an excipient
and/or the immediate container must be assessed. Depending ASSAY
on the result of this assessment, limits of degradation and/ or
Unless otherwise justified and authorised, contents of active
reaction products are set and monitored in the pharmaceutical
preparation. Licensed products require a stabílity exercise. substances and specific excipients such as preservatives are
determined in pharmaceutical preparations. Limits rnust be
Methods used for the purpose of stability testing for all defined and justified.
relevant characteristics of the preparation are validated as
Suitable and validated methods are used. If assay methods
stability indicating, i.e. the rnethods allow the quantification of prescribed in the respective active substance monographs are
the relevant degradation products and physical characteristic used, it must be demonstrated that they are not affected by the
changes. presence of the excipients and/or by the formulation.
TESTS Reference standards. See Tests.
Relevant tests to apply in order to ensure the appropriate LABELLING AND STORAGE
quality of a particular dosage form are described in the specific
dosage form monographs. The relevant labelling requirements given in the general
dosage form rnonographs apply. In addition, relevant
Where it is not practica!, for unlicensed pharrnaceutical European Union or other applícable regulations apply.
preparations, to carry out the tests (e.g. batch size, time
restraints), other suitable methods are implernented to ensure GLOSSARY
that the appropriate quality is achieved in accordance with the Formulation: the designing of an appropriate formula
risk assessment carried out and any local guidance or legal (including materials, processes, etc.) that will ensure that the
requirements. patient receives the suitable pharmaceutical preparation in an
Stock preparations are normally tested to a greater extent than appropriate forro that has the required quality and that will be
extemporaneous preparatíons. stable and effective for the required length of time.

EUROPEAN PHARMACOPOEIA 9.5 Vaccines for human use
Licensed pharmaceutical preparation: a medicinal product Bacterial vaccines containing bacterial components are
that has been granted a marketing authorisation by a suspensions or freeze-dried products. They may be adsorbed.
competent authority. Synonym: authorised pharmaceutical The antigen content is determined by a suitable validated assay.
preparation. Bacterial toxoids are prepared from toxins by diminishing their
Manufacture: all operations of purchase of materials and toxicity to an acceptable level or by completely eliminating
products, Production, Quality Control, release, storage, it by physical or chemical procedures whilst retaining
distribution of medicinal products and the related controls. adequate immunogenic properties. The toxins are obtained
Preparation (of an unlicensed pharmaceutical from selected strains of micro-organisms. The method of
preparation): the 'manufacture' of unlicensed pharmaceutical production is such that the toxoid <loes not revert to toxin.
preparations by or at the request of pharmacies or other The toxoids are purified. Purification is perfarmed befare
healthcare establishments (the term 'preparation' is used and/or after detoxification. Toxoid vaccines may be adsorbed.
instead of 'manufacture' in order clearly to distinguish it Viral vaccines are prepared from viruses grown in animals, in
from the industrial manufacture of licensed pharmaceutical fertilised eggs, in suitable cell cultures or in suitable tissues, or
preparations). by culture of genetically engineered cells. They are liquids that
Reconstitution: manipulation to enable the use or application vary in opacity according to the type of preparation or may
of a medicinal product with a marketing authorisation in be freeze-dried. They may be adsorbed. Liquid preparations
accordance with the instructions given in the summary of and freeze-dried preparations after reconstitution may be
product characteristics or the patient infarmation leaflet. coloured if a pH indicator such as phenol red has been used
in the culture medium.
Risk assessment: the identification of hazards and the analysis
and evaluation of risks associated with exposure to those Synthetic antigen vaccines are generally clear or colourless
hazards. liquids. The concentration of the components is usually
expressed in terms of specific antigen content.
Unlicensed pharmaceutical preparation: a medicinal
product that is exempt from the need of having a marketing Combined vaccines are multicomponent preparations
authorisation issued by a competent authority but is made far farmulated so that different antigens are administered
specific patients' needs according to legislation. simultaneously. The different antigenic components are
intended to protect against different strains or types of
the same organism and/or against different organisms. A
combined vaccine may be supplied by the manufacturer either
07/2018:0153 as a single liquid or freeze-dried preparation or as several
constituents with directions far admixture befare use. Where
there is no monograph to cover a particular combination, the
vaccine complies with the monograph far each individual
component, with any necessary modifications approved by
VACCINES FOR HUMAN USE the competent authority.
Adsorbed vaccines are suspensions and may farm a sediment
Vaccina ad usum humanum at the bottom of the container.
DEFINITION PRODUCTION
Vaccines far human use are preparations containing antigens
General provisions. The production method far a given
capable of inducing a specific and active immunity in man
product must have been shown to yield consistently batches
against an infecting agent or the toxin or antigen elaborated
comparable with the batch of proven clinical efficacy,
by it. Immune responses include the induction of the innate
immunogenicity and safety in man. Product specifications
and the adaptive (cellular, humoral) parts of the immune
including in-process testing should be set. Specific
system. Vaccines far human use shall have been shown to have
requirements far production including in-process testing
acceptable immunogenic activity and safety in man with the
are included in individual monographs. Where justified and
intended vaccination schedule.
authorised, certain tests may be omitted where it can be
Vaccines far human use may contain: whole micro-organisms demonstrated, far example by validation studies, that the
(bacteria, viruses or parasites), inactivated by chemical production process consistently ensures compliance with the
or physical means that maintain adequate immunogenic test.
properties; whole live micro-organisms that are naturally
avirulent or that have been treated to attenuate their virulence Unless otherwise justified and authorised, vaccines are
whilst retaining adequate immunogenic properties; antigens produced using a seed-lot system. The methods of preparation
extracted from the micro-organisms or secreted by the are designed to maintain adequate immunogenic properties, to
micro-organisms or produced by genetic engineering or render the preparation harmless and to prevent contamination
chemical synthesis. The antigens may be used in their native with extraneous agents.
state or may be detoxified or otherwise modified by chemical Where vaccines far human use are manufactured using
or physical means and may be aggregated, polymerised or materials of human or animal origin, the general requirements
conjugated to a carrier to increase their immunogenicity. of general chapter 5.1.7. Viral safety apply in conjunction with
Vaccines may contain an adjuvant. Where the antigen is the more specific requirements relating to viral safety in this
adsorbed on a mineral adjuvant, the vaccine is referred to as monograph, in general chapters 5.2.2. Chicken flocks free from
'adsorbed'. specified pathogens for the production and quality control of
Terminology used in monographs on vaccines far human use vaccines, 5.2.3. Cell substrates for the production of vaccines
is defined in general chapter 5.2.1. for human use and 2.6.16. Tests for extraneous agents in viral
vaccines for human use, and in individual monographs.
Bacterial vaccines containing whole cells are suspensions of
various degrees of opacity in colourless or almost colourless Unless otherwise justified and authorised, in the production of
liquids, or may be freeze-dried. They may be adsorbed. The a final lot of vaccine, the number of passages of a virus, or the
concentration of living or inactivated bacteria is expressed in number of subcultures of a bacterium, from the master seed
terms of International Units of opacity or, where appropriate, lot shall not exceed that used far production of the vaccine
is determined by direct cell count or, far live bacteria, by shown to be satisfactory in clinical trials with respect to safety
viable count. and efficacy or immunogenicity.
Vaccines for human use EUROPEAN PHARMACOPOEIA 9.5
Vaccines are as far as possible free from ingredients known to potential extraneous agents. A test for effectiveness of the
cause toxic, allergic or other undesirable reactions in man. inactivation process is carried out as soon as possible after
Suitable additives, including stabilisers and adjuvants may be the inactivation process.
incorporated. Penicillin and streptomycin are neither used Carrier proteins. Bacterial polysaccharide antigens may
at any stage of production nor added to the final product; be conjugated with carrier proteins to improve their
however, master seed lots prepared with media containing immunogenicity to enable the induction of a protective
penicillin or streptomycin may, where justified and authorised, response in infants. Carrier proteins comply with the relevant
be used for production. requirements of general chapter 5.2.11. Carrier proteins for the
Consistency of production is an important feature of vaccine production of conjugated polysaccharide vaccines for human
production. Monographs on vaccines for human use give use.
limits for various tests carried out during production and on
Test for sterility of intermediates prior to final bulk.
the final lot. These limits may be in the form of maximum
Individual monographs on vaccines for human use may
values, minimum values, or minimum and maximum
prescribe a test for sterility for intermediates.
tolerances around a given value. While compliance with these
limits is required, it is not necessarily sufficient to ensure In agreement with the competent authority, replacement of
consistency of production for a given vaccine. For relevant the sterility test by a bioburden test with a low bioburden limit
tests, the manufacturer must therefore define for each product based on batch data and process validation may be acceptable
for intermediates preceding the final bulk, provided that a
a suitable action or release limit or limits to be applied in view
of the results found for batches tested clinically and those sterilising filtration is performed later in the production
used to demonstrate consistency of production. These limits process.
may subsequently be refined on a statistical basis in light of It is a prerequisite that the intermediate is filtered through
production data. a bacteria-retentive filter prior to storage, that authorised
Substrates for propagation. Substrates for propagation pre-filtration bioburden limits have been established for this
comply with the relevant requirements of the Pharmacopoeia filtration, and that adequate measures are in place to avoid
(5.2.2, 5.2.3) or in the absence of such requirements with contamination and growth of micro-organisms during storage
those of the competent authority. Processing of cell banks and of the intermediate.
subsequent cell cultures is done under aseptic conditions in Final bulk. The final bulk is prepared by aseptically blending
an area where no other cells are being handled. Serum and the ingredients of the vaccine. For non-liquid vaccines for
trypsin used in the preparation of cell suspensions shall be administration by a non-parenteral route, the final bulk is
shown to be free from extraneous agents. prepared by blending the ingredients of the vaccine under
Seed lots/ cell banks. The master seed lot or cell bank is suitable conditions.
identified by historical records that include information on Adjuvants. One or more adjuvants may be included in the
its origin and subsequent manipulation. Suitable measures formulation of a vaccine to potentiate and/ or modulate
are taken to ensure that no extraneous agent or undesirable the immune response to the antigen(s). Adjuvants may be
substance is present in a master or working seed lot or a cell included in the formulation of the final vaccine or presented
bank. separately. Suitable characterisation and quality control of the
Culture media. Culture media are as far as possible free from adjuvant(s), alone and in combination with the antigen(s),
is essential for consistent production. Quality specifications
ingredients known to cause toxic, allergic or other undesirable
reactions in man; if inclusion of such ingredients is necessary,are established for each adjuvant, alone and in combination
it shall be demonstrated that the amount present in the final with the antigen(s).
lot is reduced to such a level as to render the product safe. Adsorbents as adjuvants. Vaccines may be adsorbed on
Approved animal (but not human) serum may be used in the aluminium hydroxide, aluminium phosphate, calcium
growth medium for cell cultures but the medium used for phosphate or other suitable adsorbents. The adsorbents are
maintaining cell growth during virus multiplication shall not prepared in special conditions that confer the appropriate
contain serum, unless otherwise stated. Cell culture media physical form and adsorptive properties.
may contain a pH indicator such as phenol red and approved Where an adsorbent is used as an adjuvant and is generated
antibiotics at the lowest effective concentration, although it in situ during production of the vaccine, quality specifications
is preferable to have a medium free from antibiotics during are established for each of the ingredients and for the
production. generated adsorbent in the vaccine. Quality specifications are
Propagation and harvest. The seed cultures are propagated intended to control, in particular:
and harvested under defined conditions. The purity of - qualitative and quantitative chemical composition;
the harvest is verified by suitable tests as defined in the - physical form and associated adsorptive properties, where
monograph. relevant, and particularly where the adjuvant will be
Control cells. For vaccines produced in cell cultures, control present as an adsorbent;
cells are maintained and tested as prescribed. In order to - interaction between adjuvant and antigen;
provide a valid control, these cells must be maintained in - purity, including bacteria! endotoxin content and
conditions that are essentially equivalent to those used for the microbiological quality;
production cell cultures, including use of the same batches
of media and media changes. - any other parameters identified as being critical for
functionality.
Control eggs. For live vaccines produced in eggs, control eggs The stability of each adjuvant, alone and in combination
are incubated and tested as prescribed in the monograph. with the antigen(s), particularly for critical parameters, is
Purification. Where applicable, validated purification established during development studies.
procedures may be applied. Antimicrobial preservatives. Antimicrobial preservatives
Inactivation. Inactivated vaccines are produced using are used to prevent spoilage or adverse effects caused by
a validated inactivation process whose effectiveness and microbial contamination occurring during the use of a
consistency have been demonstrated. Where it is recognised vaccine. Antimicrobial preservatives are not included in
that extraneous agents may be present in a harvest, for freeze-dried products. For single-dose liquid preparations,
example in vaccines produced in eggs from healthy, non-SPF inclusion of antimicrobial preservatives is not normally
flocks, the inactivation process is also validated with respect acceptable. For multidose liquid preparations, the need for
to a panel of model extraneous agents representative of the effective antimicrobial preservation is evaluated taking into

EUROPEAN PHARMACOPOEIA 9.5 Vaccines for human use
account likely contamination during use and the maximum freeze-drying, potency at release and real-time stability under
recommended period of use after broaching of the container. the prescribed conditions as well as thermal stability. Where
If an antimicrobial preservative is used, it shall be shown there is a significant change in the manufacturing procedure
that it <loes not impair the safety or efficacy of the vaccine. of the antigen(s) or formulation, the need for re-introduction
Addition of antibiotics as antimicrobial preservatives is not of the test is considered.
normally acceptable. Stability. During development studies, maintenance of
During development studies, the effectiveness of the potency of the final lot throughout the period of validity shall
antimicrobial preservative throughout the period of validity be demonstrated; the loss of potency in the recommended
shall be demonstrated to the satisfaction of the competent storage conditions is assessed. Excessive loss even within the
authority. limits of acceptable potency may indicate that the vaccine is
The efficacy of the antimicrobial preservative is evaluated as unacceptable.
described in general chapter 5.1.3. If neither the A criteria nor Expiry date. Unless otherwise stated, the expiry date is
the B criteria can be met, then in justified cases the following calculated from the beginning of the assay or from the
criteria are applied to vaccines for human use: bacteria, no beginning of the first assay for a combined vaccine. Por
increase at 24 h and 7 days, 3 log 10 reduction at 14 days, no vaccines stored at a temperature lower than that used for
increase at 28 days; fungí, no increase at 14 days and 28 days. stability studies and intended for release without re-assay, the
Stability of intermediates. During production of vaccines, expiry date is calculated from the date of removal from cold
intermediates are obtained at various stages and are stored, storage. If, for a given vaccine, an assay is not carried out, the
sometimes for long periods. Such intermediates include: expiry date for the final lot is calculated from the date of an
approved stability-indicating test or, failing this, from the date
- seed lots and cell banks; of freeze-drying or the date of filling into the final containers.
- live or inactivated harvests; For a combined vaccine where components are presented in
- purified harvests that may consist of toxins or toxoids, separate containers, the expiry date is that of the component
polysaccharides, bacteria! or viral suspensions; which expires first.
- purified antigens; The expiry date applies to vaccines stored in the prescribed
- adsorbed antigens; conditions.
- conjugated polysaccharides; Animal tests. In accordance with the provisions of the
- final bulk vaccine; European Convention for the Protection of Vertebrate
Animals Used for Experimental and Other Scientific Purposes,
- vaccine in the final closed container stored at a temperature tests must be carried out in such a way as to use the mínimum
lower than that used for final-product stability studies and number of animals and to cause the least pain, suffering,
intended for release without re-assay. distress or lasting harm. The criteria for judging tests in
Except where they are used within a short period of time, monographs must be applied in light of this. For example,
stability studies are carried out on the intermediates in the if it is indicated that an animal is considered to be positive,
intended storage conditions to establish the expected extent of infected, etc. when typical clinical signs or death occur,
degradation. For final bulk vaccine, stability studies may be then as soon as sufficient indication of a positive result is
carried out on representative samples in conditions equivalent obtained the animal in question shall be either euthanised or
to those intended to be used for storage. For each intermediate given suitable treatment to prevent unnecessary suffering. In
a
(except for seed lots and cell banks)) period of validity ~ccordance with the Gener~l Notices, alternative test methods
applicable for the intended storage conditions is established, may be used to demonstrate compliance with the monograph
where appropriate in light of stability studies. and the use of such tests is particularly encouraged when this
Final lot. The final lot is prepared by aseptically distributing leads to replacement or reduction of animal use or reduction
the final bulk into sterile, tamper-proof containers, which, of suffering. Guidance on how to substitute in vivo methods
after freeze-drying where applicable, are closed so as to exclude by in vitro methods, in cases where a direct head-to-head
contamination. For non-liquid vaccines for administration by comparison is not possible, can be found in general chapter
a non-parenteral route, the final lot is prepared by distributing 5.2.14.
the final bulk under suitable conditions into sterile,
TESTS
tamper-proof containers. Where justified and authorised,
certain tests prescribed for the final lot may be carried out on Vaccines comply with the tests prescribed in individual
the final bulk, if it has been demonstrated that subsequent monographs including, where applicable, the following:
manufacturing operations do not affect compliance. pH (2.2.3). Liquid vaccines, after reconstitution where
Appearance. Unless otherwise justified and authorised, applicable, comply with the limits for pH approved for the
each container (vial, syringe or ampoule) in each final lot is particular preparation.
inspected visually or mechanically for acceptable appearance. Adjuvant. If the vaccine contains an adjuvant, the amount
Degree of adsorption. For an adsorbed vaccine, unless is determined and shown to be within acceptable limits
otherwise justified and authorised, a release specification with respect to the expected amount (see also the tests for
for the degree of adsorption is established in light of results aluminium and calcium below).
found for batches used in clinical trials. From the stability Aluminium (2.5.13): maximum 1.25 mg of aluminium (Al)
data generated for the vaccine it must be shown that at the per single human <lose where an aluminium adsorbent has
end of the period of validity the degree of adsorption is not been used in the vaccine, unless otherwise stated.
less than for batches used in clinical trials.
Cakium (2.5.14): maximum 1.3 mg of calcium (Ca) per
Thermal stability. When the thermal stability test is prescribed single human <lose where a calcium adsorbent has been used
in a monograph for a live attenuated vaccine, the test is carried in the vaccine, unless otherwise stated.
out on the final lot to monitor the lot-to-lot consistency in
heat-sensitivity of viral/bacterial particles in the product. Free formaldehyde (2.4.18): maximum 0.2 g/L of free
Suitable conditions are indicated in the individual monograph. formaldehyde in the final product where formaldehyde has
The test may be omitted as a routine test for a given product been used in the preparation of the vaccine, unless otherwise
once the consistency of the production process has been stated.
demonstrated, in agreement with the competent authority, Phenol (2.5.15): maximum 2.5 g/L in the final product where
using relevant parameters, such as consistency in yield, phenol has been used in the preparation of the vaccine, unless
ratio of infectious viruses (viable bacteria) befare and after otherwise stated.
Vaccines for veterinary use EUROPEAN PHARMACOPOEIA 9.5
Water (2.5.12): maximum 3.0 per cent m/m for freeze-dried retaining all or part of their antigenic properties; vaccines may
vaccines, unless otherwise stated. also contain combinations of these constituents. The antigens
Extractable volume (2.9.17). Unless otherwise justified and may be produced by recombinant DNA technology. Suitable
authorised, it complies with the requirement for extractable adjuvants may be included to enhance the immunising
volume. properties of the vaccines.
Terminology used in monographs on vaccines for veterinary
Bacteria} endotoxins. Unless otherwise justified and
use is defined in general chapter 5.2.1.
authorised, a test for bacteria! endotoxins is carried out on the
final product. Where no limit is specified in the individual 1-1. BACTERIAL VACCINES AND BACTERIAL TOXOIDS
monograph, the content of bacteria! endotoxins determined Bacteria! vaccines and bacteria! toxoids are prepared from
by a suitable method (2. 6.14) is less than the limit approved cultures grown on suitable solid or liquid media, or by other
for the particular product. suitable means; the requirements of this section do not
apply to bacteria! vaccines prepared in cell cultures or in live
STORAGE animals. The strain ofbacterium used may have been modified
Store protected from light. Unless otherwise stated, the by genetic engineering. The identity, antigenic potency and
storage temperature is 5 ± 3 ºC; liquid adsorbed vaccines must purity of each bacteria! culture used is carefully controlled.
not be allowed to freeze. Bacteria! vaccines contain inactivated or live bacteria or their
antigenic components; they are liquid preparations of various
LABELLING
degrees of opacity or they may be freeze-dried.
The label states :
Bacteria! toxoids are prepared from toxins by diminishing
- the name of the preparation; their toxicity to a very low level or by completely eliminating
- a reference identifying the final lot; it by physical or chemical means whilst retaining adequate
- the recommended human <lose and route of administration; immunising potency. The toxins are obtained from selected
- the storage conditions; strains of specified micro-organisms grown in suitable media
or are obtained by other suitable means, for example, chemical
- the expiry date;
synthesis.
- the name and amount of any antimicrobial preservative;
The toxoids may be:
- the name of any antibiotic, adjuvant, flavour or stabiliser
- liquid;
present in the vaccine;
where applicable, that the vaccine is adsorbed; - precipitated with alum or another suitable agent;
the name of any constituent that may cause adverse - purified and/or adsorbed on aluminium phosphate,
reactions and any contra-indications to the use of the aluminium hydroxide, calcium phosphate or another
vaccine; adsorbent prescribed in the monograph.
- for freeze-dried vaccines: Bacteria! toxoids are clear or slightly opalescent liquids.
Adsorbed toxoids are suspensions or emulsions. Certain
- the name or composition and the volume of the toxoids may be freeze-dried.
reconstituting liquid to be added;
Unless otherwise indicated, statements and requirements
- the time within which the vaccine is to be used after given below for bacteria! vaccines apply equally to bacteria!
reconstitution. vaccines, bacteria! toxoids and products containing a
combination of bacteria! cells and toxoid.
07/2018:0062
1-2. VIRAL VACCINES
Viral vaccines are prepared by growth in suitable cell cultures
(5.2.4), in tissues, in micro-organisms, in fertilised eggs or,
where no other possibility is available, in live animals, or
VACCINES FOR VETERINARY USE by other suitable means. The strain of virus used may have
been modified by genetic engineering. They are liquid or
freeze-dried preparations of one or more viruses or viral
Vaccina ad usum veterinarium subunits or peptides.
In the case of combined vaccines, for each component that is the Live viral vaccines are prepared from viruses of attenuated
subject of a monograph in the Pharmacopoeia, the provisions virulence or of natural low virulence for the target species.
of that monograph apply to that component, modified where
necessary as indicated (see general chapters 5.2.6. Evaluation Inactivated viral vaccines are treated by a validated procedure
of safety of veterinary vaccines and immunosera and 5.2.7. for inactivation of the virus and may be purified and
Evaluation of efficacy of veterinary vaccines and immunosera). concentrated.
If an immunological product for veterinary use is in tended for 1-3. VECTOR VACCINES
minor use, certain tests may be excluded, subject to approval Vector vaccines are liquid or freeze-dried preparations of
by the competent authorityOJ. one or more types of live micro-organisms (bacteria, viruses
or fungí) that are non-pathogenic or have low pathogenicity
l. DEFINITION for the target species and in which have been inserted one
Vaccines for veterinary use are preparations containing or more genes encoding antigens that stimulate an immune
antigenic substances and are administered for the purpose response protective against other micro-organisms.
of inducing a specific and active immunity against disease
provoked by bacteria, toxins, viruses, fungí or parasites. The 2. PRODUCTION
vaccines, live or inactivated, confer active immunity that General provisions. Production is designed to provide
may be transferred passively via maternal antibodies against a finished product that complies with the approved
the immunogens they contain and sometimes also against requirements. Compliance with these requirements is
antigenically related organisms. Vaccines may contain living demonstrated by safety and efficacy studies carried out on
or inactivated micro-organisms (bacteria, viruses or fungi), batches during development and by the control strategy.
parasites, or antigenic fractions or substances produced The tests to be applied are outlined below and in individual
by these organisms (e.g. toxins), rendered harmless whilst monographs. In accordance with the General Notices,
(1) NOTE: Guideline on data requirements for immunological veterinary medicinal products intended for minor use or minor species/limited markets (EMA/CVMP/IWP/123243/2006,
including any subsequent revision of this document).

EUROPEAN PHARMACOPOEIA 9.5 Vaccines for veterinary use
performance of all the tests in a monograph is not necessarily 2-1-3-1-1. General requirements. The genus and species
a prerequisite for a manufacturer in assessing compliance with (and varieties where appropriate) of the bacteria used in
the Pharmacopoeia before release of a product. Therefore, the vaccine are stated. Bacteria used in manufacture are
routinely used in vivo tests can ultimately be replaced in handled in a seed-lot system wherever possible. Each master
accordance with the principies of the European Convention for seed lot is tested as described below. A record of the origin,
the Protection of Vertebrate Animals used for Experimental date of isolation, passage history (including purification
and Other Scientific Purposes, if the product profile is well and characterisation procedures) and storage conditions is
defined by a set of parameters, including antigen content and maintained for each master seed lot. Each master seed lot is
antigen quality, established to verify that the manufacturing assigned a specific code for identification purposes.
process consistently produces final batches equivalent to a final 2-1-3-1-2. Propagation. The minimum and maximum
batch that fulfils the criteria of the European Pharmacopoeia. number of subcultures of each master seed lot prior to the
production stage are specified. The methods used for the
2-1. STARTING MATERIAL preparation of seed cultures, preparation of suspensions
Monographs prescribe a set of measures that, taken together, for seeding, techniques for inoculation of seeds, titre and
give an acceptable degree of assurance that the final product concentration of inocula and the media used, are documented.
<loes not contain infectious extraneous agents. These measures lt shall be demonstrated that the characteristics of the seed
include: material (for example, dissociation or antigenicity) are not
l. production within a seed-lot system anda cell-seed system, changed by these subcultures. The conditions under which
where applicable; each seed lot is stored are documented.
2. extensive testing of seed lots and cell seed for extraneous 2-1-3-1-3. Identity and purity. Each master seed lot is shown
agents; to contain only the species and strain of bacterium stated. A
brief description of the method of identifying each strain by
3. requirements for SPF flocks used for providing substrates biochemical, serological and morphological characteristics
for vaccine production; and distinguishing it as far as possible from related strains is
4. testing of substances of animal origin, which must, wherever recorded, as is also the method of determining the purity of
possible, undergo an inactivation procedure. the strain. If the master seed lot is shown to contain living
Substances of animal origin used in the production of vaccines organisms of any kind other than the species and strain stated,
for veterinary use comply with the requirements of general then it is unsuitable for vaccine production.
chapter 5.2.5. Other substances used in the preparation of 2-1-3-2. Virus seed lots
vaccines for veterinary use comply with requirements of the 2-1-3-2-1. General requirements. Viruses used in manufacture
Pharmacopoeia (where a relevant monograph exists) and are handled in a seed-lot system. Each master seed lot is tested
are prepared in a manner that avoids contamination of the as described below. A record of the origin, date of isolation,
vaccine. passage history (including purification and characterisation
2-1-1. Substrates for production. Cell cultures used in the procedures) and storage conditions is maintained for each
production of vaccines for veterinary use comply with the seed lot. Each master seed lot is assigned a specific code for
requirements of general chapter 5.2.4. identification purposes. Production of vaccine is not normally
undertaken using virus more than 5 passages from the master
Where a monograph refers to chicken flocks free from
seed lot In the tests on the master seed lot described below,
specified pathogens (SPF), these flocks comply with the
the organisms used are not normally more than 5 passages
requirements prescribed in general chapter 5.2.2.
from the master seed lot at the start of the tests, unless
For production of inactivated vaccines, where vaccine otherwise indicated.
organisms are grown in embryonated hens' eggs, such eggs are Where the master seed lot is contained within a permanently
derived either from SPF flocks (5.2.2) or from healthy non-SPF infected master cell seed, the following tests are carried out
flocks (5.2.13). lt may be necessary to demonstrate that the on an appropriate volume of virus from disrupted master cell
inactivation process is effective against specified potential seed. Where relevant tests have been carried out on disrupted
contaminants. For the production of a master seed lot and cells to validate the suitability of the master cell seed, these
for all passages of a micro-organism up to and including the tests need not be repeated.
working seed lot, eggs from SPF flocks (5.2.2) are used.
2-1-3-2-2. Propagation. The master seed lot and all
Where it is unavoidable to use animals or animal tissues in the subsequent passages are propagated on cells, on embryonated
production of veterinary vaccines, such animals shall be free eggs or in animals that have been shown to be suitable for
from specified pathogens, as appropriate to the source species vaccine production (see above), and, where applicable, using
and the target animal for the vaccine. substances of animal origin that meet the requirements
2-1-2. Media used for seed culture preparation and for prescribed in general chapter 5.2.5.
production. Media used for seed culture preparation and for 2-1-3-2-3. Identification. A suitable method to identify the
production are prepared following a standard formulation. vaccine strain and to distinguish it as far as possible from
The media composition is described in the manufacturing related strains must be used.
process. The qualitative and quantitative composition of all 2-1-3-2-4. Bacteria and fungi. The master seed lot complies
media used must be recorded. Where media or ingredients with the test for sterility (2. 6.1).
are claimed as proprietary, this is indicated and an appropriate
description recorded. Ingredients that are derived from 2-1-3-2-5. Mycoplasmas (2.6.7). The master seed lot complies
animals are specified as to the source species and country of with the test for mycoplasmas (culture method and indicator
origin, and must comply with the criteria described in general cell culture method).
chapter 5.2.5. Preparation processes for media used, including 2-1-3-2-6. Absence of extraneous viruses. Monographs may
sterilisation procedures, are documented. contain requirements for freedom from extraneous agents,
otherwise the requirements stated below apply.
The addition of antibiotics during the manufacturing
process is normally restricted to cell culture fluids and other Preparations of monoclonal or polyclonal antibodies
media, egg inocula and material harvested from tissues and containing high levels of neutralising antibody to the virus
embryonated eggs. of the seed lot are made on a batch basis, using antigen that
is not derived from any passage level of the virus isolate
2-1-3. Seed lots giving rise to the master seed virus. Each batch of serum is
2-1-3-1. Bacteria[ seed lots maintained at 56 ºC for 30 min to inactivate complement.
Each batch is shown to be free of antibodies to potential 2-2-1-2. Information for performing the safety and efficacy
contaminants of the seed virus and is shown to be free of studies.
any non-specific inhibiting effects on the ability of viruses to During development of a vaccine, safety and immunogenicity
infect and propagate within cells (or eggs, where applicable). are demonstrated for each mute and for each method of
If such a serum cannot be obtained, other methods are used administration to be recommended. The following is a
to remove or neutralise the seed virus specifically. non-exhaustive list of such mutes of administration:
If the seed lot virus would interfere with the conduct and - intramuscular;
sensitivity of a test for extraneous viruses, a sample of the - subcutaneous;
master seed lot is treated with a minimum amount of the - intravenous;
monoclonal or polyclonal antibody so that the vaccine
- ocular;
virus is neutralised as far as possible or removed. The final
virus-serum mixture shall, if possible, contain at least the virus - oral;
content of 10 doses of vaccine per 0.1 mL for avian vaccines - nasal;
and per millilitre for other vaccines. For avian vaccines, the - foot-stab;
testing to be carried out on seed lots is given in general chapter
- wing web;
2.6.24. For mammalian vaccines, the seed lot or the mixture of
seed lot and antiserum is tested for freedom from extraneous - intradermal;
agents as follows. - intraperitoneal;
- in ovo.
The mixture is inoculated onto cultures of at least 70 cm 2 of
the required cell types. The cultures may be inoculated at any The following is a non-exhaustive list of such methods of
suitable stage of growth up to 70 per cent confluency. At least administration:
1 monolayer of each type must be retained as a control. The - injection;
cultures must be monitored daily for a week. At the end of - drinking water;
this period the cultures are freeze thawed 3 times, centrifuged - spray;
to remove cell debris and re-inoculated onto the same cell
type as above. This is repeated twice. The final passage must - eye-drop;
produce sufficient cells in appropriate vessels to carry out the - scarification;
tests below. - implantation;
Cytopathic and haemadsorbing agents are tested for using - immersion.
the methods described in the relevant sections on testing cell Monographs may indicate that a given test is to be carried
cultures (5.2.4) and techniques such as immunofluorescence out for each category of animal of the target species for which
are used for detection of specific contaminants for the tests in the product is recommended or is to be recommended. The
cell cultures. The master seed lot is inoculated onto: following is a non-exhaustive list of categories that are to be
taken into account.
- primary cells of the species of origin of the virus; Mammals:
cells sensitive to viruses pathogenic for the species for - pregnant animals/non-pregnant animals;
which the vaccine is intended; - animals raised primarily for breeding/animals raised
primarily for food production;
- cells sensitive to pestiviruses.
animals of the minimum age or size recommended for
If the master seed lot is shown to contain living organisms of vaccination.
any kind, other than the virus of the species and strain stated, - A vian species:
or foreign viral antigens, then it is unsuitable for vaccine - birds raised primarily for egg production/birds raised
production. primarily for production of meat;
2-2. CHOICE OF VACCINE COMPOSITION AND CHOICE - birds before point of lay/birds after onset of lay.
OF VACCINE STRAIN - Fish:
For the choice of vaccine composition and choice of vaccine - broodstock fish/fish raised primarily for food
strain, important aspects to be evaluated include safety, production.
efficacy and stability.
2-2-2. Antimicrobial preservatives. Antimicrobial
2-2-1. Development studies on safety and efficacy. General preservatives are used to prevent spoilage or adverse effects
requirements for evaluation of safety and efficacy are given in caused by microbial contamination occurring during use of a
general chapters 5.2. 6 and 5.2. 7. These requirements may be vaccine which is expected to be no longer than 1O h after first
made more explicit or supplemented by the requirements of broaching. Antimicrobial preservatives are not included in
individual monographs. freeze-dried products but, if justified, taking into account the
maximum recommended period of use after reconstitution,
2-2-1-1. Potency and immunogenicity. The tests given under they may be included in the diluent for multi-dose
the headings Potency and Immunogenicity in monographs freeze-dried products. For single-dose liquid preparations,
serve 2 purposes: inclusion of antimicrobial preservatives is not acceptable
- the Potency section establishes, by a well-controlled test unless justified and authorised, but may be acceptable, for
in experimental conditions, the minimum acceptable example where the same vaccine is filled in single-dose and
vaccinating capacity for all vaccines within the scope of multidose containers and is used in non-food-producing
the definition, which must be guaranteed throughout the species. For multidose liquid preparations, the need for
period of validity; effective antimicrobial preservation is evaluated taking into
account likely contamination during use and the maximum
- well-controlled experimental studies are normally a part recommended period of use after broaching of the container.
of the overall demonstration of efficacy of a vaccine During development studies the effectiveness of the
(see general chapter 5.2. 7); the test referred to in the antimicrobial preservative throughout the period of validity
Immunogenicity section (to which the Potency section shall be demonstrated to the satisfaction of the competent
usually cross-refers) is suitable as a part of this testing. authority.

The efficacy of the antimicrobial preservative is evaluated as given production process. The rest of this section applies to
described in general chapter 5.1.3 and in addition samples are each production run. When conducting tests for inactivation,
tested at suitable intervals over the proposed in-use shelf-life. it is essential to take account of the possibility that under
If neither the A criteria nor the B criteria can be met, then in the conditions of manufacture, organisms may be physically
justified cases the following criteria are applied to vaccines protected from inactivant.
for veterinary use: bacteria, no increase from 24 h to 7 days, 2-3-2-1. Inactivation kinetics. The inactivating agent and
3 log 10 reduction at 14 days, no increase at 28 days; fungi, no the inactivation procedure shall be shown, under conditions
increase at 14 days and 28 days. of manufacture, to inactivate the vaccine micro-organism.
Addition of antibiotics as antimicrobial preservative is Adequate data on inactivation kinetics shall be obtained.
generally not acceptable. Normally, the time required for inactivation shall be not more
2-2-3. Stability. Evidence of stability is obtained to justify than 67 per cent of the duration of the inactivation process.
the proposed period of validity. This evidence takes the form The maximum titre of the vaccine micro-organism capable of
of the results of virus titrations, bacteria! counts or potency being inactivated by the selected method is established based
tests carried out at regular intervals until 3 months beyond on the inactivation kinetics data.
the end of the shelf life on not fewer than 3 representative 2-3-2-2. Residues of inactivating agents. Appropriate tests are
consecutive batches of vaccine kept under recommended carried out to demonstrate that the inactivating agent has
storage conditions together with results from studies of been removed or reduced to an acceptable residual level.
moisture content (for freeze-dried products), physical tests on If an aziridine compound is used as the inactivating agent, this
the adjuvant, chemical tests on substances such as the adjuvant may be accomplished by neutralising it with thiosulfate and
constituents and preservatives, and pH, as appropriate. demonstrating residual thiosulfate in the inactivated harvest
Where applicable, studies on the stability of the reconstituted at the completion of the inactivation procedure.
vaccine are carried out, using the product reconstituted in If formaldehyde is used as the inactivating agent, then a test
accordance with the proposed recommendations. for free formaldehyde is carried out as prescribed under
The variations in the results obtained during the stability study 3. Batch tests.
are taken into account when defining appropriate formulation 2-3-2-3. Residual live virus/bacteria and!or detoxification
and release specifications to ensure the conformity of the testing. A test for complete inactivation and/or detoxification
product for the claimed shelf-life. is performed immediately after the inactivation and/or
2-2-4. Formulation. The minimum antigen content, virus detoxification procedure and, if applicable, the neutralisation
titre or bacterial count acceptable from the point of view of or removal of the inactivating or the test for detoxifying agent.
efficacy (i.e. gives satisfactory results in the potency test and Validation of the test for residual live virus/bacteria or the test
other efficacy studies) is established during development for detoxification shall focus on the level of detection of the
studies. The antigen formulation, where applicable the live virus/bacteria or toxin.
adjuvant formulation, and the release specifications are set 2-3-2-3-1. Bacteria! vaccines. The test selected shall be
based on this minimum value and based on the results of the appropriate to the vaccine bacteria being used and shall
stability studies. consist of at least 2 passages in production medium or, if solid
A maximum antigen content, virus titre or bacteria! count, medium has been used for production, in a suitable liquid
acceptable from the point of view o[ safety, is established medium or in the medium prescribed in the monograph.
during development studies. The product complies with the test if no evidence of any live
micro-organism is observed.
For live vaccines, this is also used as the maximum acceptable
titre for each batch of vaccine at release. 2-3-2-3-2. Bacteria! toxoids. The test selected shall be
appropriate to the toxin or toxins present and shall be the
2-3. PREPARA TION OF THE VACCINE most sensitive available.
The methods of preparation, which vary according to
the type of vaccine, are such as to maintain the integrity 2-3-2-3-3. Viral vaccines. The test selected shall be appropriate
and immunogenicity of the antigen, to ensure freedom to the vaccine virus being used and must consist of at least
from contamination with extraneous agents and to ensure 2 passages in cells, embryonated eggs or, where no other
production of vaccine batches of consistent quality. suitably sensitive method is available, in animals. The quantity
For each individual product, relevant in-process and finished of cell samples, eggs or animals shall be sufficient to ensure
product controls are established to verify the production appropriate sensitivity of the test. For tests in cell cultures, not
process and the batch-to-batch quality of the product. The less than 150 cm 2 of cell culture monolayer is inoculated with
results are within the approved limits defined for the particular 1.0 mL of inactivated harvest. The product complies with the
product. test if no evidence of the presence of any live virus or other
micro-organism is observed.
2-3-1. Propagation and harvest of bacteria! and viral
antigens. Each strain of a multivalent vaccine is cultivated 2-3-3. Final bulk and final batch. The final bulk vaccine is
and harvested separately. prepared by combining one or more batches of antigen, that
comply with all the relevant requirements, with any auxiliary
The working seed materials are propagated in suitable substances, such as adjuvants, stabilisers, antimicrobial
media/substrates for production. The conditions of these preservatives and diluents.
propagation steps are described and monitored by recording
appropriate parameters, e.g. temperature, pH, duration, The vaccine is blended according to a defined formulation.
turbidity and oxygen saturation. The results are within the Unless otherwise prescribed in the individual monograph
approved limits defined for the particular product. or otherwise justified and authorised, the final bulk vaccine
During production, where possible, growth rate is monitored is distributed aseptically with or without freeze-drying, into
by suitable methods and the values are recorded and within sterile, tamper-proof containers, which are then closed so as
the approved limits defined for the particular product. The to prevent contamination. This constitutes the final batch.
antigen may then be inactivated and/or purified and/or 2-4. MANUFACTURER'S TESTS
concentrated. 2-4-1. Antigen content. The formulation of the vaccine is
2-3-2. Inactivation. Inactivated vaccines are subjected based, whenever possible, on the antigen content determined
to a validated inactivation procedure. The testing of the on the harvest before or after inactivation and/or downstream
inactivation kinetics described below is carried out once for a processing, if applicable.
2-4-2. Batch potency test. Por most vaccines, the tests cited shall be either euthanised or given suitable treatment to
under Potency or Immunogenicity are not suitable for the prevent unnecessary suffering. In accordance with the General
routine testing of batches. Notices, alternative test methods may be used to demonstrate
If the test described under Potency is not used for routine compliance with the monograph and the use of such tests is
testing, a batch potency test is established during development. particularly encouraged when this leads to replacement or
The aim of the batch potency test is to ensure that each batch of reduction of animal use or reduction of suffering. Guidance
vaccine would, if tested, comply with the test described under on how to substitute in vivo methods by in vitro methods, in
Potency and Immunogenicity. The acceptance criteria for the cases where a direct head-to-head comparison is not possible,
batch potency test are therefore established by correlation with can be found in general chapter 5.2.14.
the test described under Potency. Where a batch potency test is Taking into account the quality systems in place and advances
described in a monograph, this is given as an example of a test in scientific knowledge and understanding of the products,
that is considered suitable, after establishment of correlation manufacturing processes and their controls, the choice of
with the potency test; other test models can also be used. tests to be performed may be reconsidered when assessing
Por live vaccines, virus titre or bacteria! count is generally compliance with Pharmacopoeia monographs, in accordance
appropriate as a batch potency test. with the General Notices. On a case-by-case basis, with
the agreement of the competent authority, the choice and
Por inactivated vaccines, development of in vitro methods is necessity of certain final product tests may be reconsidered,
recommended, provided that: where in-process tests give at least an equal guarantee that
- key in-process parameters are defined and monitored; the batch would comply if tested, or where alternative tests
- in-process control tests (including antigen quantification validated with respect to the Pharmacopoeia method have
after inactivation and/or concentration, if applicable) and been carried out.
target formulation of the final product are performed. All hen eggs, chickens and chicken cell cultures for use in
Antigen content. The quantity of appropriate antigen per <lose, quality control tests shall be derived from an SPP flock (5.2.2).
determined by a suitable method, is not significantly lower 3-1. Identification. The antigen is identified by suitable
than that of a batch of vaccine that has given satisfactory methods such as nucleic acid amplification techniques
results in the test described under Potency. (2.6.21). Por inactivated vaccines, the identification test may
Adjuvant. If the test for antigen content is performed and be combined with the batch potency test.
if the vaccine is adjuvanted, the identity of the adjuvant is 3-2. Physical tests. A vaccine with an oily adjuvant is tested
verified by suitable chemical methods and the adjuvant is for viscosity by a suitable method and shown to be within the
tested as described in section 3. Batch tests. The quality and limits set for the product. The stability of the emulsion shall
quantity of the adjuvant is not significantly different from that be demonstrated.
of a batch of vaccine that has given satisfactory results in the
3-3. Chemical tests. Tests for the concentrations of
test described under Potency.
appropriate substances such as aluminium and preservatives
2-5. IN-PROCESS STABILITY are carried out to show that these are within the limits set for
During production of vaccines, intermediate products are the product.
obtained at various stages and may be stored. The intended 3-4. pH. The pH ofliquid products and diluents is measured if
conditions and duration of storage are defined in light of the possible and shown to be within the limits set for the product.
stability data.
3-5. Water. Where applicable, the freeze-drying process is
3. BATCH TESTS checked by a determination of water and shown to be within
the limits set for the product.
The individual monographs also indicate tests to be carried out
on each particular vaccine. 3-6. Formaldehyde (2.4.18; use Method B if sodium
metabisulfite has been used to neutralise excess
Certain tests may be carried out on the final bulk vaccine formaldehyde). Where formaldehyde has been used in the
rather than on the batch or batches prepared from it; such preparation, the concentration of free formaldehyde is not
tests include those for antimicrobial preservatives, free greater than 0.5 g/L, unless a higher amount has been shown
formaldehyde and the potency determination for inactivated to be safe.
vaccines.
3-7. Phenol (2.5.15). When the vaccine contains phenol, the
Under particular circumstances (i.e. significant changes to
concentration is not greater than 5 g/L.
the manufacturing process, as well as reports of unexpected
adverse reactions observed in the field or reports that the 3-8. Bacteria and fungi. Vaccines comply with the test for
final batches do not comply with the former data provided sterility (2.6.1). Where the volume of liquid in a container
during licensing), other tests, including tests on animals, is greater than 100 mL, the method of membrane filtration
may be needed on an ad hoc basis; they are carried out in is used wherever possible. Where the method of membrane
agreement with or at the request of the competent authority. filtration cannot be used, the method of direct inoculation
Por safety testing, one or more of the tests described in general may be used. Where the volume of liquid in each container
chapter 5.2.6 may be carried out. is at least 20 mL, the mínimum volume to be used for each
culture medium is 1O per cent of the contents or 5 mL,
Only a batch that complies with each of the requirements
whichever is less. The appropriate number of items to be
given below, completed or amended by the requirements given
tested (2. 6.1) is 1 per cent of the batch with a mínimum of 4
in the relevant individual monograph, may be released for use.
anda maximum of 10.
Animal tests. In accordance with the provisions of the
Por live bacteria! and for live fungal vaccines, the absence of
European Convention for the Protection of Vertebrate
Animals Used for Experimental and Other Scientific Purposes, micro-organisms other than the vaccine strain is demonstrated
by suitable methods such as microscopic examination and
tests must be carried out in such a way as to use the mínimum
inoculation of suitable media.
number of animals and to cause the least pain, suffering,
distress or lasting harm. The criteria for judging tests in Por frozen or freeze-dried avian live viral vaccines produced
monographs must be applied in light of this. Por example, in embryonated eggs, for non-parenteral use only, the
if it is indicated that an animal is considered to be positive, requirement for sterility is usually replaced by requirements
infected etc. when typical clinical signs occur then as soon as for absence of pathogenic micro-organisms and for a
it is clear that result will not be affected the animal in question maximum of 1 non-pathogenic micro-organism per <lose.

3-9. Extraneous agents. Avian live viral vaccines comply with Expiry date. Unless otherwise stated, the expiry date is .
the tests for extraneous agents in batches of finished product calculated from the beginning of the virus titration or bactenal
(2.6.25). count (for live vaccines) or the beginning of the potency test
Mammalian live viral vaccines are tested for extraneous (for other vaccines). For combined vaccines, the expiry date
viruses using an appropriate method. is that of the component which expires first. For vaccines
stored by the manufacturer at a temperature lower than that
In case of doubt, the tests intended for the seed lot of a stated on the label, the stability for the entire storage period
live vaccine may also be applied to the final product. If an is demonstrated by an appropriate study. The expiry date is
extraneous agent is found in such a test, the vaccine <loes not then calculated from the date that the vaccine is stored in the
comply with the monograph. conditions stated on the label.
3-1 O. Residual live virus/bacteria and/ or detoxification The expiry date applies to vaccines stored in the prescribed
testing. For inactivated vaccines and bacteria! toxoids, the conditions.
tests described in section 2-3-2-3 are performed. Where
auxiliary substances would interfere with a test for inactivation5. LABELLING
and/or detoxification, such a test is carried out during The label states:
preparation of the final bulk, after the different batches of - that the preparation is for veterinary use;
antigen have been combined, but before addition of the
- the volume of the preparation and the number of doses in
auxiliary substances; the test for inactivation or detoxification
the container;
may then be omitted on the final bulk and the final batch.
- the mute of administration;
The test for residual live virus/bacteria may be omitted for
batch release provided that: - the type or types of bacteria (and where applicable the
antigenic components) or viruses used and for live vaccines
1. a titration is performed on each harvest prior to inactivation the minimum and the maximum number of live bacteria
and the titre is not greater than the maximum titre or the minimum and the maximum virus titre;
established based on studies of the inactivation kinetics;
- where applicable, for inactivated vaccines, the minimum
2. suitable test sensitivity for residual live virus/bacteria has potency in International Units;
been demonstrated; - where applicable, the name of any antimicrobial
3. the test for residual live virus/bacteria is performed with preservative or other substance added to the vaccine;
satisfactory results on each harvest. - the name of any substance that may cause an adverse
Where there is a risk of reversion to toxicity, the test for reaction;
detoxification performed at the latest stage of the production - for freeze-dried vaccines:
process at which the sensitivity of the test is not compromised - the name or composition and the volume of the
(e.g. after the different batches of antigen have been combined reconstituting liquid to be added;
but before the addition of auxiliary substances) is important
to demonstrate a lack of reversion to toxicity. - the period within which the vaccine is to be used after
reconstitution;
3-11. Mycoplasmas (2.6.7). Live viral vaccines comply with - for vaccines with an oily adjuvant, that if the vaccine is
the test for mycoplasmas (culture method). accidentally injected into man, urgent medical attention
~- 1? Potencv Tht> v;:icdne comnlies with the reauirements is necessary;
~f-th.e -t~;t-~-~~ti~~~d-~~der Im~unogenicity (se~ section - the animal species for which the vaccine is intended;
2-2-1-1) when administered by a recommended route and
- the indications for the vaccine;
method.
- the instructions for use;
4. STORAGE - any contra-indications to the use of the product including
Store protected from light at a temperature of 5 ± 3 ºC, unless any required warning on the dangers of administration of
otherwise indicated. Liquid preparations are not to be allowed an overdose;
to freeze, unless otherwise indicated. - the doses recommended for different species.


Diphtheria, tetanus, pertussis (acellular, component) and Diphtheria, tetanus, pertussis (whole cell), poliomyelitis
haemophilus type b conjugate vaccine (adsorbed) .......... 5583 (inactivated) and haemophilus type b conjugate vaccine
Diphtheria, tetanus, pertussis (acellular, component), (adsorbed) ............................................................................. 5590
hepatitis B (rDNA), poliomyelitis {inactivated) and Haemophilus type b conjugate vaccine ................................ 5592
haemophilus type b conjugate vaccine (adsorbed) .......... 5585
Diphtheria, tetanus, pertussis (acellular, component),
poliomyelitis (inactivated) and haemophilus type b conjugate
vaccine (adsorbed) ............................................................... 5587

EUROPEAN PHARMACOPOEIA 9.5 DIP- TET- PERª-HIB
07/2018:1932 inject subcutaneously 5 times the single human <lose stated on

the label into each of 5 healthy guinea-pigs, each weighing
250-350 g, that have not previously been treated with any
material that will interfere with the test. If within 42 days of
the injection any of the animals shows signs of or dies from
diphtheria toxaemia or tetanus, the vaccine <loes not comply
DIPHTHERIA, TETANUS, PERTUSSIS with the test. If more than 1 animal dies from non-specific
(ACELLULAR, COMPONENT) AND causes, repeat the test once; if more than 1 animal dies in the
second test, the vaccine <loes not comply with the test.
HAEMOPHILUS TYPE b CONJUGATE
The content of bacteria! endotoxins (2. 6.14) in bulk purified
VACCINE (ADSORBED) diphtheria toxoid, tetanus toxoid, pertussis components and
bulk PRP conjugate is determined to monitor the purification
Vaccinum diphtheriae, tetani, pertussis procedure and to limit the amount in the final vaccine. Por
each component, the content of bacteria! endotoxins is less
sine cellulis ex elementis praeparatum et than the limit approved for the particular vaccine; where the
haemophili stirpis b coniugatum adsorbatum haemophilus component is presented in a separate container,
the contents of the diphtheria, tetanus and pertussis antigens
DEPINITION are in any case such that the final vial for these components
Diphtheria, tetanus, pertussis (acellular, component) and contains less than 100 IU per single human <lose.
haemophilus type b conjugate vaccine (adsorbed) is a The production method is validated to demonstrate that the
combined vaccine composed of: diphtheria formol toxoid; product, if tested, would comply with the test for abnormal
tetanus formol toxoid; individually purified antigenic toxicity for immunosera and vaccines for human use (2.6.9).
components of Bordetella pertussis; polyribosylribitol During development studies, it shall be demonstrated that
phosphate (PRP) covalently bound to a carrier protein; a the vaccine consistently induces a T-cell-dependent B-cell
mineral absorbent such as aluminium hydroxide or hydrated immune response to PRP. If the manufacturing process is
aluminium phosphate. The product is presented either as a modified, it shall be demonstrated by appropriate in vitro
tetravalent liquid formulation in the same container, or as a methods that the characteristic properties of the conjugate
trivalent liquid formulation with the haemophilus component are not affected.
in a separate container, the contents of which are mixed with
the other components immediately before use. Where the haemophilus component is presented in a separate
container, the production method is validated to demonstrate
The formol toxoids are prepared from the toxins produced by that the haemophilus component, if tested, would comply
the growth of Corynebacterium diphtheriae and Clostridium with the test for pyrogens (2.6.8), carried out as follows: inject
tetani respectively. per kilogram of the rabbit's mass a quantity of the vaccine
The vaccine contains either pertussis toxoid or a equivalent to: 1 µg of PRP for a vaccine with diphtheria toxoid
pertussis-toxin-like protein free from toxic properties or CRM 197 diphtheria protein as carrier; 0.1 µg of PRP for a
produced by expression of a genetically modified form of vaccine with tetanus toxoid as carrier; 0.025 µg of PRP for a
the corresponding gene. Pertussis toxoid is prepared from vaccine with OMP (meningococcal group B outer membrane
pertussis toxin by a method that rcnders the toxin harmless nrntPin rnmnlpv) ""'U<>e r<irriPr
.t'.L'-.J\.-..L..1...1. -'-'.L.&..1.l"..1.-...1.11../ -"4.L..1..1.-.&.•
while maintaining adequate immunogenic properties Reference vaccine(s). Provided valid assays can be performed,
and avoiding reversion to toxin. The acellular pertussis monocomponent reference vaccines may be used for the
component may also contain filamentous haemagglutinin, assays on the combined vaccine. If this is not possible because
pertactin (a 69 kDa outer-membrane protein) and other of interaction between the components of the combined
defined components of B. pertussis such as fimbrial-2 and vaccine or because of differences in composition between
fimbrial-3 antigens. The latter 2 antigens may be co-purified. the monocomponent reference vaccine and the test vaccine,
The antigenic composition and characteristics are based a batch of combined vaccine shown to be effective in clinical
on evidence of protection and freedom from unexpected trials or a batch representative thereof is used as a reference
reactions in the target group for which the vaccine is intended. vaccine. Por the preparation of a representative batch, strict
PRP is a linear copolymer composed of repeated units adherence to the production process used for the batch tested
of 3-~-D-ribofuranosyl-(1..¿ 1)-ribitol-5-phosphate in clinical trials is necessary. The reference vaccine may be
[(C 10 H 1p 12 P)J, with a defined molecular size and derived stabilised by a method that has been shown to have no effect
from a suitable strain of Haemophilus influenzae type b. on the assay procedure.
The carrier protein, when conjugated to PRP, is capable of PRODUCTION OF THE COMPONENTS
inducing a T-cell-dependent B-cell immune response to the The production of the components complies with the
polysaccharide. requirements of the monographs Diphtheria vaccine
(adsorbed) (0443), Tetanus vaccine (adsorbed) (0452),
PRODUCTION Pertussis vaccine (acellular, component, adsorbed) (1356) and
GENERAL PROVISIONS Haemophilus type b conjugate vaccine (1219).
The production method shall have been shown to yield FINAL BULK VACCINE
consistently vaccines comparable with the vaccine of proven Different methods of preparation may be used: a final bulk
clinical efficacy and safety in man. vaccine may be prepared by adsorption, separately or together,
Where the haemophilus component is presented in a separate of suitable quantities of bulk purified diphtheria toxoid,
container, as part of consistency studies the assays of the tetanus toxoid, acellular pertussis components and PRP
diphtheria, tetanus and pertussis components are carried out conjugate onto a mineral carrier such as aluminium hydroxide
on a suitable number of batches of vaccine reconstituted as or hydrated aluminium phosphate; or 2 final bulks may be
for use. For subsequent routine control, the assays of these prepared and filled separately, one containing the diphtheria,
components may be carried out without mixing with the tetanus and pertussis components, the other the haemophilus
haemophilus component. component, which may be freeze-dried. Suitable antimicrobial
Specific toxicity of the diphtheria and tetanus components. preservatives may be added.
The production method is validated to demonstrate that Only a final bulk vaccine that complies with the following
the product, if tested, would comply with the following test: requirements may be used in the preparation of the final lot.
DIP-TET-PERª -HIB EUROPEAN PHARMACOPOEIA 9.5
Antimicrobial preservative. Where applicable, determine the If the haemophilus component is freeze-dried, sorne tests may
amount of antimicrobial preservative by a suitable chemical be carried out on the freeze-dried product rather than on the
method. The amount is not less than 85 per cent and not bulk conjugate where the freeze-drying process may affect the
greater than 115 per cent of the intended content. component to be tested.
Sterility (2.6.1). Carry out the test for sterility using 10 mL Residual pertussis toxin and irreversibility of pertussis
for each medium. toxoid (2.6.33). The final lot complies with the test.
FINAL LOT PRP: minimum 80 per cent of the amount of PRP stated on the
Only a final lot that is satisfactory with respect to the test label. PRP is determined either by assay of ribose (2.5.31) or
for osmolality shown below and with respect to each of the phosphorus (2.5.18), by an immunochemical method (2.7.1)
requirements given below under Identification, Tests and or by anion-exchange liquid chromatography (2.2.29) with
Assay may be released for use. pulsed amperometric detection.
Provided the test for residual pertussis toxin and irreversibility Aluminium (2.5.13): maximum 1.25 mg per single human
of pertussis toxoid, the test for antimicrobial preservative and <lose, if aluminium hydroxide or hydrated aluminium
the assay have been carried out with satisfactory results on the phosphate is used as the adsorbent.
final bulk vaccine, they may be omitted on the final lot. Free formaldehyde (2.4.18): maximum 0.2 g/L.
Provided the free formaldehyde content has been determined Antimicrobial preservative. Where applicable, determine the
on the bulk purified antigens or the final bulk and it has been amount of antimicrobial preservative by a suitable chemical
shown that the content in the final lot will not exceed 0.2 g/L, method. The content is not less than the minimum amount
the test for free formaldehyde may be omitted on the final lot. shown to be effective and is not greater than 115 per cent of
Osmolality (2.2.35). The osmolality of the vaccine, the quantity stated on the label.
reconstituted where applicable, is within the limits approved Water (2.5.12): maximum 3.0 per cent for the freeze-dried
for the particular preparation. haemophilus component.
pH (2.2.3). The pH of the vaccine, reconstituted if necessary, Sterility (2.6.1). lt complies with the test for sterility.
is within the range approved for the particular product.
Bacteria} endotoxins (2.6.14). The content is within the limits
Free PRP. Unbound PRP is determined after removal of the approved by the competent authority for the haemophilus
conjugate, for example by anion-exchange, size-exclusion component of the particular product. If any components of
or hydrophobic chromatography, ultrafiltration or other the vaccine prevent the determination of endotoxin, a test for
validated methods. The amount of free PRP is not greater pyrogens is carried out as described under General provisions.
than that approved for the particular product.
ASSAY
IDENTIFICATION Diphtheria component. Carry out one of the prescribed
Where the haemophilus component is presented in a separate methods for the assay of diphtheria vaccine (adsorbed) (2.7.6).
container: identification tests A, B and C are carried out using The lower confidence limit (P = 0.95) of the estimated potency
the container containing the diphtheria, tetanus and pertussis is not less than the minimum potency stated on the label.
components; identification test D is carried out on the container
containing the haemophilus component. Unless otherwise justified and authorised, the minimum
potency stated on the label is 30 IU per single human <lose.
A. Diphtheria toxoid is identified by a suitable
immunochemical method (2.7.1). The following Tetanus component. Carry out one of the prescribed methods
method, applicable to certain vaccines, is given as an for the assay of tetanus vaccine (adsorbed) (2.7.8).
example. Dissolve in the vaccine to be examined sufficient The lower confidence limit (P = 0.95) of the estimated potency
sodium citrate R to give a 100 g/L solution. Maintain at is not less than 40 IU per single human dose.
37 ºC for about 16 h and centrifuge until a clear supernatant
Pertussis component. Carry out one of the prescribed
is obtained. The clear supernatant reacts with a suitable
methods for the assay of pertussis vaccine (acellular) (2.7.16).
diphtheria antitoxin, giving a precipitate.
The capacity of the vaccine to induce antibodies for each
B. Tetanus toxoid is identified by a suitable immunochemical
included acellular pertussis antigen is not significantly
method (2.7.1). The following method, applicable to certain
(P = 0.95) less than that of the reference vaccine.
vaccines, is given as an example. The clear supernatant
obtained as described in identification test A reacts with a
LABELLING
suitable tetanus antitoxin, giving a precipitate.
The label states:
C. The pertussis components are identified by a suitable
immunochemical method (2.7.1). The following method, - the minimum number of International Units of diphtheria
applicable to certain vaccines, is given as an example. The and tetanus toxoid per single human <lose;
clear supernatant obtained as described in identification - the names and amounts of the pertussis components per
test A reacts with specific antisera to the pertussis single human dos e;
components of the vaccine.
- the number of micrograms of PRP per single human <lose;
D. The haemophilus component is identified by a suitable
immunochemical method (2. 7.1) for PRP. - the type and nominal amount of carrier protein per single
human <lose;
TESTS - where applicable, that the vaccine is intended for primary
vaccination of children and is not necessarily suitable for
Where the product is presented with the haemophilus reinforcing doses or for administration to adults;
component in a separate container: the tests for residual
pertussis toxin and irreversibility of pertussis toxoid, - the name and the amount of the adsorbent;
aluminium, free formaldehyde, antimicrobial preservative and - that the vaccine must be shaken before use;
sterility are carried out on the container with the diphtheria,
tetanus and pertussis components; the tests for PRP content, - that the vaccine is not to be frozen;
water (where applicable), sterility and bacterial endotoxins are - where applicable, that the vaccine contains a pertussis
carried out on the container with the haemophilus component. toxin-like protein produced by genetic modification.

EUROPEAN PHARMACOPOEIA 9.5 DIP-TET-PER3 -HBV-IPV-HIB
07 /2018:2067 If the vaccine is presented with the haemophilus component in

a separate container, as part of consistency studies the assays of
the diphtheria, tetanus, pertussis, hepatitis B and poliomyelitis
components are carried out on a suitable number of batches
of vaccine reconstituted as for use. For subsequent routine
DIPHTHERIA, TETANUS, PERTUSSIS control, the assays of these components may be carried out
without mixing with the haemophilus component.
(ACELLULAR, COMPONENT),
Specific toxicity of the diphtheria and tetanus components.
HEPATITIS B (rDNA), POLIOMYELITIS The production method is validated to demonstrate that
(INACTIVATED) AND HAEMOPHILUS the product, if tested, would comply with the following test:
inject subcutaneously 5 times the single human <lose stated on
TYPE b CONJUGATE VACCINE the label into each of 5 healthy guinea-pigs, each weighing
(ADSORBED) 250-350 g, that have not previously been treated with any
material that will interfere with the test. If within 42 days of
Vaccinum diphtheriae, tetani, pertussis the injection any of the animals shows signs of or dies from
diphtheria toxaemia or tetanus, the vaccine <loes not comply
sine cellulis ex elementis praeparatum, with the test. If more than 1 animal dies from non-specific
hepatitidis B (ADNr), poliomyelitidis causes, repeat the test once; if more than 1 animal dies in the
inactivatum et haemophili stirpis b second test, the vaccine <loes not comply with the test.
The content of bacteria} endotoxins (2.6.14) in bulk purified
coniugatum adsorbatum diphtheria toxoid, tetanus toxoid and pertussis components,
DEFINITION hepatitis B surface antigen, purified, inactivated monovalent
Diphtheria, tetanus, pertussis (acellular, component), poliovirus harvests and bulk PRP conjugate is determined to
hepatitis B (rDNA), poliomyelitis (inactivated) and monitor the purification procedure and to limit the amount in
haemophilus type b conjugate vaccine (adsorbed) is a the final vaccine. For each component, the content of bacteria}
combined vaccine composed of: diphtheria formol toxoid; endotoxins is not greater than the limit approved.
tetanus formol toxoid; individually purified antigenic During development studies and wherever revalidation is
components of Bordetella pertussis; hepatitis B surface necessary, a test for pyrogens in rabbits (2.6.8) is carried out
antigen (HBsAg) ; human poliovirus types 1, 2 and 3 grown by injection of a suitable <lose of the final lot. The vaccine is
in suitable cell cultures and inactivated by a suitable method; shown to be acceptable with respect to absence of pyrogenic
polyribosylribitol phosphate (PRP) covalently bound to a activity.
carrier protein. The antigens in the vaccine may be adsorbed During development studies, it shall be demonstrated that
on a mineral carrier such as aluminium hydroxide or hydrated the vaccine consistently induces a T-cell-dependent B-cell
aluminium phosphate. The product is presented either as immune response to PRP. If the manufacturing process is
a hexavalent liquid formulation in the same container, or modified, it shall be demonstrated by appropriate in vitro
as a pentavalent liquid formulation with the haemophilus methods that the characteristic properties of the conjugate
component in a separate container, the contents of which are not affected.
d~rt;;g-~;e~·---- ---- ------
::irP mixPrl with thP othPr comnonents immediatelv
-----r -------- ---- - '
hefore or
The stability of the final lot and relevant intermediates
is evaluated using one or more indicator tests. For
The formol toxoids are prepared from the toxins produced by the haemophilus component, such tests may include
the growth of Corynebacterium diphtheriae and Clostridium determination of molecular size, determination of free
tetani respectively. PRP in the conjugate and kinetics of depolymerisation.
The vaccine contains either pertussis toxoid or a Taking account of the results of the stability testing, release
pertussis-toxin-like protein free from toxic properties requirements are set for these indicator tests to ensure that the
produced by expression of a genetically modified form of vaccine will be satisfactory at the end of the period of validity.
the corresponding gene. Pertussis toxoid is prepared from
Reference vaccine(s). Provided valid assays can be performed,
pertussis toxin by a method that renders the toxin harmless
monocomponent reference vaccines may be used for the
while maintaining adequate immunogenic properties
assays on the combined vaccine. If this is not possible because
and avoiding reversion to toxin. The acellular pertussis
of interaction between the components of the combined
component may also contain filamentous haemagglutinin,
vaccine or because of differences in composition between
pertactin (a 69 kDa outer-membrane protein) and other
the monocomponent reference vaccine and the test vaccine,
defined components of B. pertussis such as fimbrial-2 and
a batch of combined vaccine shown to be effective in clinical
fimbrial-3 antigens. The latter 2 antigens may be co-purified.
trials or a batch representative thereof is used as a reference
The antigenic composition and characteristics are based
vaccine. For the preparation of a representative batch, strict
on evidence of protection and freedom from unexpected
adherence to the production process used for the batch tested
reactions in the target group for which the vaccine is intended.
in clinical trials is necessary. The reference vaccine may be
Hepatitis B surface antigen is a component protein of stabilised by a method that has been shown to have no effect
hepatitis B virus; the antigen is obtained by recombinant on the assay procedure.
DNA technology.
PRODUCTION OF THE COMPONENTS
PRP is a linear copolymer composed of repeated units of 3-~
D-ribofuranosyl-(l ~ l)-ribitol-5-phosphate [(C 10 H 19 0 12 P)J, The production of the components complies with the
with a defined molecular size and derived from a suitable strain requirements of the monographs Diphtheria vaccine
of Haemophilus influenzae type b. The carrier protein, when (adsorbed) (0443), Tetanus vaccine (adsorbed) (0452), Pertussis
conjugated to PRP, is capable of inducing a T-cell-dependent vaccine (acellular, component, adsorbed) (1356), Hepatitis B
B-cell immune response to the polysaccharide. vaccine (rDNA) (1056), Poliomyelitis vaccine (inactivated)
(0214) and Haemophilus type b conjugate vaccine (1219).
PRODUCTION FINAL BULKS
GENERAL PROVISIONS Vaccine with all components in the same container. The final
The production method shall have been shown to yield bulk is prepared by adsorption, separately or together, of
consistently vaccines comparable with the vaccine of proven suitable quantities of bulk purified diphtheria toxoid, tetanus
clinical efficacy and safety in man. toxoid, acellular pertussis components and hepatitis B surface
DIP-TET-PERª -HBV -IPV -HIB EUROPEAN PHARMACOPOEIA 9.5
antigen onto a mineral carrier such as aluminium hydroxide same result as the in vivo assay in terms of acceptance or
or hydrated aluminium phosphate and admixture of a suitable rejection of a batch. This demonstration must include testing
quantity of PRP conjugate and suitable quantities of purified of subpotent batches, produced experimentally if necessary,
and inactivated, monovalent harvests of human poliovirus for example by heat treatment or other means of diminishing
types l, 2 and 3 or a suitable quantity of a trivalent pool of such the immunogenic activity. Where there is a significant
monovalent harvests. Suitable antimicrobial preservatives change in the manufacturing process of the antigens or their
may be added. formulation, any impact on the in vivo and in vitro assays
Vaccine with the haemophilus component in a separate must be evaluated, and the need for revalidation considered.
container. The final bulk of diphtheria, tetanus, pertussis, Free PRP. Por vaccines with all components in the same
hepatitis B and poliovirus component is prepared by container, the free PRP content is determined on the
adsorption, separately or together, of suitable quantities of non-absorbed fraction. Unbound PRP is determined on the
bulk purified diphtheria toxoid, tetanus toxoid, acellular haemophilus component after removal of the conjugate, for
pertussis components and hepatitis B surface antigen example by anion-exchange, size-exclusion or hydrophobic
onto a mineral carrier such as aluminium hydroxide or chromatography, ultrafiltration or other validated methods.
hydrated aluminium phosphate and admixture of suitable The amount of free PRP is not greater than that approved for
quantities of purified and inactivated, monovalent harvests the particular product.
of human poliovirus types l, 2 and 3 or a suitable pool of
such monovalent harvests. This final bulk is filled separately. Bacteria! endotoxins (2.6.14): less than the limit approved
Suitable antimicrobial preservatives may be added. The final for the product concerned.
bulk of the haemophilus component is prepared by dilution of Osmolality (2.2.35). The osmolality of the vaccine,
the bulk conjugate to the final concentration with a suitable reconstituted where applicable, is within the limits approved
diluent. A stabiliser may be added. for the particular preparation.
Only final bulks that comply with the following requirements
IDENTIFICATION
may be used in the preparation of the final lot.
Bovine serum albumin. Determined on the poliomyelitis
If the vaccine is presented with the haemophilus component
in a separate container: identification tests A, B, C, D and
components by a suitable immunochemical method (2. 7.1) E are carried out using the container with the diphtheria,
after purification of the harvests and before preparation of tetanus, pertussis, hepatitis B and poliomyelitis components;
the final bulk vaccine, before addition of the adsorbent, the identification test F is carried out on the container with the
amount of bovine serum albumin is such that the content haemophilus components.
in the final vaccine will be not more than 50 ng per single
human dose. A. Diphtheria toxoid is identified by a suitable
immunochemical method (2.7.1). The following
Antimicrobial preservative. Where applicable, determine the method is given as an example. Dissolve in the vaccine to
amount of antimicrobial preservative by a suitable chemical be examined sufficient sodium citrate R to give a 100 g/L
method. The amount is not less than 85 per cent and not solution. Maintain at 37 ºC for about 16 h and centrifuge
greater than 115 per cent of the intended content. until a clear supernatant is obtained. The clear supernatant
Sterility (2.6.1). Carry out the test for sterility using 10 mL reacts with a suitable diphtheria antitoxin, giving a
for each medium. precipitate.
FINALLOT B. Tetanus toxoid is identified by a suitable immunochemical
Where the haemophilus component is in a separate container, method (2.7.1). The following method is given as
the final bulk of the haemophilus component is freeze-dried. an example. The clear supernatant obtained during
Only a final lot that is satisfactory with respect to the test identification test A reacts with a suitable tetanus antitoxin,
for osmolality shown below and with respect to each of the giving a precipitate.
requirements given below under Identification, Tests and C. The clear supernatant obtained during identification test A
Assay may be released for use. reacts with specific antisera to the pertussis components of
Provided that the test for osmolality, the test for residual the vaccine when examined by suitable immunochemical
pertussis toxin and irreversibility of pertussis toxoid, the methods (2.7.1).
test for antimicrobial preservative and the assays for the D. The hepatitis B component is identified by a suitable
diphtheria, tetanus and pertussis components have been immunochemical method (2.7.1), for example the in vitro
carried out with satisfactory results on the final bulk vaccine, assay, or by a suitable electrophoretic method (2.2.31).
they may be omitted on the final lot.
E. The vaccine is shown to contain human poliovirus types l,
Provided the free formaldehyde content has been determined 2 and 3 by a suitable immunochemical method (2.7.1),
on the bulk purified antigens and the purified monovalent such as determination of D-antigen by enzyme-linked
harvests or the trivalent pool of polioviruses or the final bulk immunosorbent assay (ELISA).
and it has been shown that the content in the final lot will not
F. The PRP and its carrier protein are identified by a suitable
exceed 0.2 g/L, the test for free formaldehyde may be omitted
immunochemical method (2.7.1).
on the final lot.
Provided that the test for bovine serum albumin has been TESTS
carried out with satisfactory results on the trivalent pool of If the product is presented with the haemophilus component
inactivated monovalent harvests of polioviruses or on the final in a separate container, the tests far residual pertussis toxin
bulk vaccine, it may be omitted on the final lot. and irreversibility of pertussis toxoid, free formaldehyde,
If an in vivo assay is used for the hepatitis B component, aluminium, antimicrobial preservative and sterility are
provided it has been carried out with satisfactory results on carried out on the container with the diphtheria, tetanus,
the final bulk vaccine, it may be omitted on the final lot. pertussis, poliomyelitis and hepatitis B components; the tests
Provided the in vivo assay for the poliomyelitis component for PRP, water, antimicrobial preservative (where applicable),
has been carried out with satisfactory results on the final bulk aluminium (where applicable) and sterility are carried out on
vaccine, it may be omitted on the final lot. the container with the haemophilus component.
The in vivo assay for the poliomyelitis component may be Sorne tests for the haemophilus component are carried out on
omitted once it has been demonstrated for a given product the freeze-dried product rather than on the bulk conjugate
and for each poliovirus type that the acceptance criteria where the freeze-drying process may affect the component to
for the D-antigen determination are such that it yields the be tested.

EUROPEAN PHARMACOPOEIA 9.5 DIP-TET-PERª-IPV-HIB
Residual pertussis toxin and irreversibility of pertussis - the nominal amount of poliovirus of each type (1, 2 and 3),
toxoid (2.6.33). The final lot complies with the test. expressed in European Pharmacopoeia Units of D-antigen,
PRP: minimum 80 per cent of the amount of PRP stated on per single human <lose;
the label, for a vaccine with the haemophilus component in a - the types of cells used for production of the poliomyelitis
separate container. and the hepatitis B components;
For a vaccine with all components in the same container: the - the number of micrograms of PRP per single human <lose;
PRP content determined on the non-absorbed fraction is not - the type and nominal amount of carrier protein per single
less than that approved for the product. human <lose;
PRP is determined either by assay of ribose (2.5.31) or - where applicable, that the vaccine is intended for primary
phosphorus (2.5.18), by an immunochemical method (2.7.1) vaccination of children and is not necessarily suitable for
or by anion-exchange liquid chromatography (2.2.29) with reinforcing doses or for administration to adults;
pulsed amperometric detection. - the name and the amount of the adsorbent;
Aluminium (2.5.13): maximum 1.25 mg per single human - that the vaccine must be shaken before use;
<lose, if aluminium hydroxide or hydrated aluminium
- that the vaccine is not to be frozen;
phosphate is used as the adsorbent.
- where applicable, that the vaccine contains a pertussis
Free formaldehyde (2.4.18): maximum 0.2 g/L of free toxin-like protein produced by genetic modification.
formaldehyde per single human <lose.
Antimicrobial preservative. Where applicable, determine the
amount of antimicrobial preservative by a suitable chemical 07 /2018:2065
method. The content is not less than the mínimum amount
shown to be effective and is not greater than 115 per cent of
the quantity stated on the label.
Water (2.5.12): maximum 3.0 per cent for the freeze-dried
haemophilus component. DIPHTHERIA, TETANUS, PERTUSSIS
Sterility (2.6.1). It complies with the test for sterility. (ACELLULAR, COMPONENT),
ASSAY POLIOMYELITIS (INACTIVATED) AND
Diphtheria component. Carry out one of the prescribed HAEMOPHILUS TYPE b CONJUGATE
methods for the assay of diphtheria vaccine (adsorbed) (2.7.6). VACCINE (ADSORBED)
The lower confidence limit (P = 0.95) of the estimated potency
is not less than the mínimum potency stated on the label.
Vaccinum diphtheriae, tetani, pertussis
Unless otherwise justified and authorised, the mínimum
potency stated on the label is 30 IU per single human <lose. sine cellulis ex elementis praeparatum,
Tetanus component. Carry out one of the prescribed methods poliomyelitidis inactivatum et haemophili
for the assay of tetanus vaccine (adsorbed) (2.7.8). stirpis b coniugatum adsorbatum
is not less than 40 IU per single human <lose. DEFINITION
Diphtheria, tetanus, pertussis (acellular, component),
Pertussis component. Carry out one of the prescribed
poliomyelitis (inactivated) and haemophilus type b conjugate
methods for the assay of pertussis vaccine (acellular) (2.7.16).
vaccine (adsorbed) is a combined vaccine composed of:
The capacity of the vaccine to induce antibodies for each diphtheria formol toxoid; tetanus formol toxoid; individually
included acellular pertussis antigen is not significantly purified antigenic components of Bordetella pertussis; suitable
(P = 0.95) less than that of the reference vaccine. strains of human poliovirus types 1, 2 and 3 grown in
Hepatitis B component. The vaccine complies with the assay suitable cell cultures and inactivated by a suitable method;
of hepatitis B vaccine (2. 7.15). polyribosylribitol phosphate (PRP) covalently bound to a
Poliomyelitis component carrier protein; a mineral adsorbent such as aluminium
hydroxide or hydrated aluminium phosphate. The product is
D-antigen con ten t. As a measure of consistency of production, presented either as a pentavalent liquid formulation in the
determine the D-antigen content for human poliovirus same container, or as a tetravalent liquid formulation with the
types l, 2 and 3 by a suitable immunochemical method (2.7.1) freeze-dried haemophilus component in a separate container,
following desorption, using a reference preparation calibrated the contents of which are mixed with the other components
in European Pharmacopoeia Units of D-antigen. For immediately before use.
each type, the content, expressed with reference to the
amount of D-antigen stated on the label, is within the limits The formol toxoids are prepared from the toxins produced by
approved for the particular product. Poliomyelitis vaccine the growth of Corynebacterium diphtheriae and Clostridium
(inactivated) BRP is calibrated in European Pharmacopoeia tetani respectively.
Units and intended for use in the assay of D-antigen. The The vaccine contains either pertussis toxoid or a
European Pharmacopoeia Unit and the International Unit are pertussis-toxin-like protein free from toxic properties
equivalent. produced by expression of a genetically modified form of
In vivo test. The vaccine complies with the in vivo assay of the corresponding gene. Pertussis toxoid is prepared from
poliomyelitis vaccine (inactivated) (2.7.20). pertussis toxin by a method that renders the toxin harmless
while maintaining adequate immunogenic properties
LABELLING and avoiding reversion to toxin. The acellular pertussis
The label states : component may also contain filamentous haemagglutinin,
pertactin (a 69 kDa outer-membrane protein) and other
- the mínimum number of International Units of diphtheria
defined components of B. pertussis such as fimbrial-2 and
and tetanus toxoid per single human <lose;
fimbrial-3 antigens. The latter 2 antigens may be co-purified.
- the names and amounts of the pertussis components per The antigenic composition and characteristics are based
single human <lose; on evidence of protection and freedom from unexpected
- the amount of HBsAg per single human <lose; reactions in the target group for which the vaccine is intended.
DIP-TET-PERª -IPV -HIB EUROPEAN PHARMACOPOEIA 9.5
PRP is a linear copolymer composed of repeated units PRODUCTION OF THE COMPONENTS

of 3-~-D-ribofuranosyl-(1 ~ l)-ribitol-5-phosphate The production of the components complies with the
[(C 10 H 1p 12 P)J, with a defined molecular size and derived requirements of the monographs Diphtheria vaccine
from a suitable strain of Haemophilus influenzae type b. (adsorbed) (0443), Tetanus vaccine (adsorbed) (0452), Pertussis
The carrier protein, when conjugated to PRP, is capable of vaccine (acellular, component, adsorbed) (1356), Poliomyelitis
inducing a T-cell-dependent B-cell immune response to the vaccine (inactivated) (0214) and Haemophilus type b conjugate
polysaccharide. vaccine (1219).
FINALBULKS
PRODUCTION The final tetravalent bulk of the diphtheria, tetanus, pertussis
GENERAL PROVISIONS and poliomyelitis components is prepared by adsorption,
The production method shall have been shown to yield separately or together, of suitable quantities of bulk purified
consistently vaccines comparable with the vaccine of proven diphtheria toxoid, bulk purified tetanus toxoid and bulk
clinical efficacy and safety in man. purified acellular pertussis components onto a mineral
carrier such as aluminium hydroxide or hydrated aluminium
Specific toxicity of the diphtheria and tetanus components. phosphate, and admixture of suitable quantities of purified,
The production method is validated to demonstrate that monovalent harvests of human poliovirus types l, 2 and 3
the product, if tested, would comply with the following test: or a suitable quantity of a trivalent pool of such monovalent
inject subcutaneously 5 times the single human <lose stated on harvests. Suitable antimicrobial preservatives may be added.
the label into each of 5 healthy guinea-pigs, each weighing
Where the vaccine is presented with all 5 components in
250-350 g, that have not previously been treated with any the same container, the final bulk is prepared by addition
material that will interfere with the test. If within 42 days of of a suitable quantity of the haemophilus bulk conjugate to
the injection any of the animals shows signs of or dies from
the tetravalent bulk. Where the haemophilus component is
diphtheria toxaemia or tetanus, the vaccine <loes not comply presented in a separate container, the final bulk is prepared
with the test. If more than 1 animal dies from non-specific
by dilution of the bulk conjugate with suitable diluents for
causes, repeat the test once; if more than 1 animal dies in the
freeze-drying. A stabiliser may be added.
second test, the vaccine <loes not comply with the test.
Only final bulks that comply with the following requirements
The content of bacteria! endotoxins (2.6.14) in bulk purified may be used in the preparation of the final lot.
diphtheria toxoid, tetanus toxoid, pertussis components,
purified, inactivated monovalent poliovirus harvests and bulk Bovine serum albumin. Determined on the poliomyelitis
PRP conjugate is determined to monitor the purification components by a suitable immunochemical method (2.7.1)
procedure and to limit the amount in the final vaccine. For during preparation of the final bulk vaccine, before addition
each component, the content of bacteria! endotoxins is less of the adsorbent, the amount of bovine serum albumin is such
than the limit approved by the competent authority for the that the contcnt in thc final vaccinc vvill be not more than
particular vaccine. 50 ng per single human <lose.
During development studies, it shall be demonstrated that amount of antimicrobial preservative by a suitable chemical
the vaccine consistently induces a T -cell-dependent B-cell method. The amount is not less than 85 per cent and not
immune response to PRP. If the manufacturing process is greater than 115 per cent of the intended content.
modified, it shall be demonstrated by appropriate in vitro
methods that the characteristic properties of the conjugate Sterility (2.6.1). Carry out the test for sterility using 10 mL
are not affected. for each medium.
Where the haemophilus component is presented in a separate FINALLOT
container, and as part of consistency studies, the assays of the Where the haemophilus component is presented in a separate
diphtheria, tetanus, pertussis and poliomyelitis components container, the final bulk of the haemophilus component is
are carried out on a suitable number of batches of vaccine freeze-dried.
reconstituted as for use. For subsequent routine control, the Only a final lot that is satisfactory with respect to the test
assays of these components may be carried out without mixing for osmolality shown below and with respect to each of the
with the haemophilus component. requirements given below under Identification, Tests and
Assay may be released for use.
Where the haemophilus component is presented in a separate
container, the production method is validated to demonstrate Provided that the test for residual pertussis toxin and
that the haemophilus component, if tested, would comply irreversibility of pertussis toxoid, the test for antimicrobial
with the test for pyrogens (2.6.8), carried out as follows: inject preservative and the assay have been carried out with
per kilogram of the rabbit's mass a quantity of the vaccine satisfactory results on the final bulk vaccine, they may be
equivalent to: 1 µg of PRP for a vaccine with diphtheria toxoid omitted on the final lot.
or CRM 197 diphtheria protein as carrier; 0.1 µg of PRP for a Provided that the free formaldehyde content has been
vaccine with tetanus toxoid as carrier; 0.025 µg of PRP for a determined on the bulk purified antigens and the purified
vaccine with OMP (meningococcal group B outer membrane monovalent harvests or the trivalent pool of polioviruses or
protein complex) as carrier. the final bulk and it has been shown that the content in the
final lot will not exceed 0.2 g/L, the test for free formaldehyde
Reference vaccine(s). Provided valid assays can be performed,
may be omitted on the final lot.
assays on the combined vaccine. If this is not possible because If the in vivo assay for the poliomyelitis component is used,
of interaction between the components of the combined provided it has been carried out with satisfactory results on
vaccine or because of differences in composition between the final bulk vaccine, it may be omitted on the final lot.
the monocomponent reference vaccine and the test vaccine, The in vivo assay for the poliomyelitis component may be
a batch of combined vaccine shown to be effective in clinical omitted once it has been demonstrated for a given product
trials or a batch representative thereof is used as a reference and for each poliovirus type that the acceptance criteria
vaccine. For the preparation of a representative batch, strict for the D-antigen determination are such that it yields the
adherence to the production process used for the batch tested same result as the in vivo assay in terms of acceptance or
in clinical trials is necessary. The reference vaccine may be rejection of a batch. This demonstration must include testing
stabilised by a method that has been shown to have no effect of subpotent batches, produced experimentally if necessary,
on the assay procedure. for example by heat treatment or other means of diminishing

EUROPEAN PHARMACOPOEIA 9.5 DIP-TET-PERª -IPV -HIB
the immunogenic activity. Where there is a significant Residual pertussis toxin and irreversibility of pertussis
change in the manufacturing process of the antigens or their toxoid (2.6.33). The final lot complies with the test.
formulation, any impact on the in vivo and in vitro assays PRP: not less than 80 per cent of the amount of PRP stated on
must be evaluated, and the need for revalidation considered. the label. PRP is determined either by assay of ribose (2.5.31)
Osmolality (2.2.35). The osmolality of the vaccine, or phosphorus (2.5.18), by an immunochemical method (2. 7.1)
reconstituted where applicable, is within the limits approved or by anion-exchange liquid chromatography (2.2.29) with
for the particular preparation. pulsed amperometric detection.
Free PRP. Where the haemophilus component is presented Aluminium (2.5.13): maximum 1.25 mg per single human
in liquid formulation, the presence of other components may dose, if aluminium hydroxide or hydrated aluminium
interfere in the assay and it may not be possible to separate phosphate is used as the adsorbent.
the PRP from the adjuvant. The presence of free PRP may Free formaldehyde (2.4.18): maximum 0.2 g/L.
be determined on the bulk conjugate prior to the addition of
other components or on the non-adsorbed fraction in the Antimicrobial preservative. Where applicable, determine the
final combination. amount of antimicrobial preservative by a suitable chemical
method. The content is not less than the mínimum amount
Where the haemophilus component is presented in a separate
shown to be effective and is not greater than 115 per cent of
container, a number of methods have been used to separate
the quantity stated on the label.
free PRP from the conjugate, including precipitation, gel
filtration, size-exclusion, anion-exchange and hydrophobic Water (2.5.12): maximum 3.0 per cent for the freeze-dried
chromatography, ultrafiltration and ultracentrifugation. The haemophilus component.
free PRP can then be quantified by a range of techniques, Sterility (2.6.1). It complies with the test for sterility.
including high-performance anion-exchange chromatography
with pulsed amperometric detection (HPAEC-PAD) and Bacteria! endotoxins (2.6.14). The content is within the limits
immunoassays with anti-PRP antibodies. The amount of approved by the competent authority for the haemophilus
free PRP is not greater than that approved for the particular component of the particular product. If any components of
product. the vaccine prevent the determination of endotoxin, a test for
pyrogens is carried out as described under General provisions.
IDENTIFICATION
ASSAY
ldentification tests A, B, C and D are carried out using the vial
containing the diphtheria, tetanus, pertussis and poliomyelitis Diphtheria component. Carry out one of the prescribed
components; identification test E is carried out either on the methods for the assay of diphtheria vaccine (adsorbed) (2.7.6).
vial containing all 5 components, or on the vial containing the Unless otherwise justified and authorised, the lower confidence
haemophilus component alone. limit (P = 0.95) of the estimated potency is not less than 30 IU
A. Diphtheria toxoid is identified by a suitable per single human dose.
immunochemical method (2.7.1). The following Tetanus component. Carry out one of the prescribed methods
method, applicable to certain vaccines, is given as an for the assay of tetanus vaccine (adsorbed) (2.7.8).
example. Dissolve in the vaccine to be examined sufficient
sodium citrate R to give a 100 g/L solution. Maintain at
is not less than 40 IU per single human dose.
37 ºC fer about 16 h and centrifuge until a clear supernatant
is obtained. The clear supernatant reacts with a suitable Pertussis component. Carry out one of the prescribed
diphtheria antitoxin, giving a precipitate. methods for the assay of pertussis vaccine (acellular) (2.7.16).
B. Tetanus toxoid is identified by a suitable immunochemical The capacity of the vaccine to induce antibodies for each
method (2.7.1). The following method, applicable to certain included acellular pertussis antigen is not significantly
vaccines, is given as an example. The clear supernatant (P = 0.95) less than that of the reference vaccine.
obtained during identification test A reacts with a suitable Poliomyelitis component
tetanus antitoxin, giving a precipitate.
D-antigen content. As a measure of consistency of production,
C. The pertussis components are identified by a suitable determine the D-antigen content for human poliovirus
immunochemical method (2.7.1). The following method, types l, 2 and 3 by a suitable immunochemical method (2.7.1)
applicable to certain vaccines, is given as an example. The following desorption, using a reference preparation calibrated
clear supernatant obtained during identification test A in European Pharmacopoeia Units of D-antigen. For
reacts with specific antisera to the pertussis components of each type, the content, expressed with reference to the
the vaccine. amount of D-antigen stated on the label, is within the limits
D. The vaccine is shown to contain human poliovirus types l, approved for the particular product. Poliomyelitis vaccine
2 and 3 by a suitable immunochemical method (2.7.1), (inactivated) BRP is calibrated in European Pharmacopoeia
such as determination of D-antigen by enzyme-linked Units and intended for use in the assay of D-antigen. The
immunosorbent assay (ELISA). European Pharmacopoeia Unit and the International Unit are
E. The haemophilus component is identified by a suitable equivalent.
immunochemical method (2.7.1) for PRP. In vivo test. The vaccine complies with the in vivo assay of
poliomyelitis vaccine (inactivated) (2.7.20).
TESTS
Where the haemophilus component is presented in a LABELLING
separate container, the tests for residual pertussis toxin The label states:
and irreversibility of pertussis toxoid, aluminium, free - the mínimum number of International Units of diphtheria
formaldehyde, antimicrobial preservative and sterility are and tetanus toxoid per single human do se;
carried out on the container with the diphtheria, tetanus,
pertussis and poliomyelitis components; the tests for PRP, - the names and amounts of the pertussis components per
water, sterility and bacteria! entodoxins are carried out on the single human dose;
container with the haemophilus component alone. - the nominal amount of poliovirus of each type (1, 2 and 3),
Where the haemophilus component is presented in a separate expressed in European Pharmacopoeia Units of D-antigen,
container, sorne tests may be carried out on the freeze-dried per single human <lose;
product rather than on the bulk conjugate where the - the type of cells used for production of the poliomyelitis
freeze-drying process may affect the component to be tested. component;
DIP- TET -PER.v-IPV -HIB EUROPEAN PHARMACOPOEIA 9.5
- the number of micrograms of PRP per single human dose;

As part of consistency studies the assays of the diphtheria,
tetanus, pertussis and poliomyelitis components are carried
- the type and nominal amount of carrier protein per single
human dose; out on a suitable number of batches of vaccine reconstituted
as for use. Por subsequent routine control, the assays of these
- where applicable, that the vaccine is intended for primary
components may be carried out without mixing with the
haemophilus component.
reinforcing doses or for administration to adults;
Por the haemophilus component, the production method is
- the name and the amount of the adsorbent;
validated to demonstrate that the haemophilus component,
- that the vaccine must be shaken before use;
if tested, would comply with the test for pyrogens (2.6.8),
- that the vaccine is not to be frozen; carried out as follows: inject per kilogram of the rabbit's
mass a quantity of the vaccine equivalent to: 1 µg of PRP
- where applicable, that the vaccine contains a
for a vaccine with diphtheria toxoid or CRM 197 diphtheria
pertussis-toxin-like protein produced by genetic
modification. protein as carrier; 0.1 µg of PRP for a vaccine with tetanus
toxoid as carrier; 0.025 µg of PRP for a vaccine with OMP
(meningococcal group B outer membrane protein complex)
as carrier.
07 /2018:2066 Reference vaccine(s). Provided valid assays can be performed,
assays on the combined vaccine. If this is not possible because
of interaction between the components of the combined
vaccine or because of the difference in composition between
DIPHTHERIA, TETANUS, PERTUSSIS monocomponent reference vaccine and the test vaccine, a
batch of combined vaccine shown to be effective in clinical
(WHOLE CELL}, POLIOMYELITIS trials or a batch representative thereof is used as a reference
(INACTIVATED) AND HAEMOPHILUS vaccine. Por the preparation of a representative batch, strict
TYPE b CONJUGATE VACCINE adherence to the production process used for the batch tested
in clinical trials is necessary. The reference vaccine may be
(ADSORBED) stabilised by a method that has been shown to have no effect
on the assay procedure.
Vaccinum diphtheriae, tetani, pertussis ex Specific toxicity of the diphtheria and tetanus components.
cellulis integris, poliomyelitidis inactivatum The production method is validated to demonstrate that
the product, if tested, would comply with the following test:
et haemophili stirpis b coniugatum inject subcutaneously 5 times the~si.ngle human dose stated on
adsorbatum the label into each of 5 healthy guinea-pigs, each weighing
250-350 g, that have not previously been treated with any
DEFINITION material that will interfere with the test. If within 42 days of
Diphtheria, tetanus, pertussis (whole cell), poliomyelitis the injection any of the animals shows signs of or dies from
(inactivated) and haemophilus type b conjugate vaccine diphtheria toxaemia or tetanus, the vaccine does not comply
(adsorbed) is a combined vaccine composed of: diphtheria with the test. If more than 1 animal dies from non-specific
formol toxoid; tetanus formol toxoid; an inactivated causes, repeat the test once; if more than 1 animal dies in the
suspension of Bordetella pertussis; suitable strains of human second test, the vaccine does not comply with the test.
poliovirus types l, 2 and 3 grown in suitable cell cultures PRODUCTION OF THE COMPONENTS
and inactivated by a suitable method; polyribosylribitol
phosphate (PRP) covalently bound to a carrier protein; a The production of the components complies with the
mineral adsorbent such as aluminium hydroxide or hydrated requirements of the monographs Diphtheria vaccine
(adsorbed) (0443), Tetanus vaccine (adsorbed) (0452), Pertussis
aluminium phosphate. The product is presented with the
vaccine (whole cell, adsorbed) (0161), Poliomyelitis vaccine
haemophilus component in a separate container, the contents
(inactivated) (0214) and Haemophilus type b conjugate
of which are mixed with the other components immediately
vaccine (1219).
before use.
The formol toxoids are prepared from the toxins produced by FINALBULKS
the growth of Corynebacterium diphtheriae and Clostridium The final bulk of the diphtheria, tetanus, pertussis and
tetani respectively. poliomyelitis components is prepared by adsorption,
separately or together, of suitable quantities of bulk purified
PRP is a linear copolymer composed of repeated units
diphtheria toxoid, and bulk purified tetanus toxoid onto a
of 3-~-D-ribofuranosyl-(1~1)-ribitol-5-phosphate
mineral carrier such as aluminium hydroxide or hydrated
[(C 10H 1p 12 P)n], with a defined molecular size and derived
aluminium phosphate and admixture of suitable quantities
from a suitable strain of Haemophilus influenzae type b.
of an inactivated suspension of B. pertussis and of purified,
The carrier protein, when conjugated to PRP, is capable of
monovalent harvests of human poliovirus types l, 2 and 3
inducing a T-cell-dependent B-cell immune response to the
or a suitable quantity of a trivalent pool of such monovalent
polysaccharide.
harvests. Suitable antimicrobial preservatives may be added.
PRODUCTION The final bulk of the haemophilus component is prepared by
GENERAL PROVISIONS dilution of the bulk conjugate to the final concentration with a
The production method shall have been shown to yield suitable diluent. A stabiliser may be added.
consistently vaccines comparable with the vaccine of proven Only final bulks that comply with the following requirements
clinical efficacy and safety in man. may be used in the preparation of the final lot.
During development studies, it shall be demonstrated that Bovine serum albumin. Determined on the poliomyelitis
the vaccine consistently induces a T -cell-dependent B-cell components by a suitable immunochemical method (2. 7.1)
immune response to PRP. If the manufacturing process is during preparation of the final bulk vaccine, before addition
modified, it shall be demonstrated by appropriate in vitro of the adsorbent, the amount of bovine serum albumin is such
methods that the characteristic properties of the conjugate that the content in the final vaccine will be not more than
are not affected. 50 ng per single human dose.

EUROPEAN PHARMACOPOEIA 9.5 DIP-TET-PEJl.v-IPV -HIB
Antimicrobial preservative. Where applicable, determine the C. The centrifugation residue obtained in identification A
amount of antimicrobial preservative by a suitable chemical may be used. Other suitable methods far separating the
method. The amount is not less than 85 per cent and not bacteria from the adsorbent may also be used. Identify
greater than 115 per cent of the intended content. pertussis vaccine by agglutination of the bacteria from the
Sterility (2.6.1). Carry out the test far sterility using 10 mL resuspended precipitate by antisera specific to B. pertussis
far each medium. or by the assay of the pertussis component prescribed
under Assay.
FINALLOT
D. The vaccine is shown to contain human poliovirus types 1,
The final bulk of the haemophilus component is freeze-dried.
2 and 3 by a suitable immunochemical method (2.7.1),
Only a final lot that is satisfactory with respect to the test such as determination of D-antigen by enzyme-linked
far osmolality shown below and with respect to each of the immunosorbent assay (ELISA).
requirements given below under Identification, Tests and E. The haemophilus component is identified by a suitable
Assay may be released far use. immunochemical method (2. 7.1) far PRP.
Provided the tests far specific toxicity of the pertussis
component and antimicrobial preservative, and the assays far TESTS
the diphtheria, tetanus and pertussis components have been
The tests for specific toxicity of the pertussis component,
carried out with satisfactory results on the final bulk vaccine,
aluminium, free formaldehyde, antimicrobial preservative
they may be omitted on the final lot.
and sterility are carried out on the container with diphtheria,
Provided the free farmaldehyde content has been determined tetanus, pertussis and poliomyelitis components; the tests for
on the bulk purified antigens, the inactivated B. pertussis PRP, water, sterility and bacteria[ endotoxins are carried out on
suspension and the purified monovalent harvests or the the container with the haemophilus component.
trivalent pool of polioviruses or on the final bulk and it has Sorne tests for the haemophilus component may be carried out
been shown that the content in the final lot will not exceed on the freeze-dried product rather than on the bulk conjugate
0.2 g/L, the test far free farmaldehyde may be omitted on the where the freeze-drying process may affect the component to
final lot. be tested.
Provided the in vivo assay far the poliomyelitis component Specific toxicity of the pertussis component. Use not fewer
has been carried out with satisfactory results on the final bulk than 5 healthy mice each weighing 14-16 g, far the vaccine
vaccine, it may be omitted on the final lot. group and far the saline control. Use mice of the same sex
The in vivo assay for the poliomyelitis component may be or distribute males and females equally between the groups.
omitted once it has been demonstrated far a given product Allow the animals access to faod and water for at least 2 h
and far each poliovirus type that the acceptance criteria befare injection and during the test. Inject each mouse of
far the D-antigen determination are such that it yields the the vaccine group intraperitoneally with 0.5 mL, containing
same result as the in vivo assay in terms of acceptance or a quantity of the vaccine equivalent to not less than half the
rejection of a batch. This demonstration must include testing single human <lose. Inject each mouse of the control group
of subpotent batches, produced experimentally if necessary, with 0.5 mL of a 9 g/L sterile solution of sodium chloride R,
far example by heat treatment or other means of diminishing preferably containing the same amount of antimicrobial
the immunogenic activity. Where there is a significant preservative as that injected with the vaccine. Weigh the
change in the manufacturing process of the antigens or their groups of mice immediately befare the injection and 72 h and
farmulation, any impact on the in vivo and in vitro assays 7 days after the injection. The vaccine complies with the test if:
must be evaluated, and the need far revalidation considered. (a) at the end of 72 h the total mass of the group of vaccinated
mice is not less than that preceding the injection; (b) at the
Osmolality (2.2.35). The osmolality of the vaccine, end of 7 days the average increase in mass per vaccinated
reconstituted where applicable, is within the limits approved mouse is not less than 60 per cent of that per control mouse;
far the particular preparation. and (c) not more than 5 per cent of the vaccinated mice die
Free PRP. Unbound PRP is determined on the haemophilus during the test. The test may be repeated and the results of
component after removal of the conjugate, far example the tests combined.
by anion-exchange, size-exclusion or hydrophobic PRP: minimum 80 per cent of the amount of PRP stated on the
chromatography, ultrafiltration or other validated methods. label. PRP is determined either by assay of ribose (2.5.31) or
The amount of free PRP is not greater than that approved far phosphorus (2.5.18), by an immunochemical method (2.7.1)
the particular product. or by anion-exchange liquid chromatography (2.2.29) with
pulsed amperometric detection.
IDENTIFICATION
Aluminium (2.5.13): maximum 1.25 mg per single human
Identification tests A, B, C and D are carried out using the vial <lose, if aluminium hydroxide or hydrated aluminium
containing the diphtheria, tetanus, pertussis and poliomyelitis phosphate is used as the adsorbent.
components; identification test E is carried out on the vial Free formaldehyde (2.4.18): maximum 0.2 g/L.
containing the haemophilus component.
A. Diphtheria toxoid is identified by a suitable amount of antimicrobial preservative by a suitable chemical
immunochemical method (2.7.1). The fallowing method. The content is not less than the minimum amount
method, applicable to certain vaccines, is given as an shown to be effective and is not greater than 115 per cent of
example. Dissolve in the vaccine to be examined sufficient the quantity stated on the label.
sodium citrate R to give a 100 g/L solution. Maintain at
37 ºC far about 16 h and centrifuge until a clear supernatant Water (2.5.12): maximum 3.0 per cent far the haemophilus
is obtained. The clear supernatant reacts with a suitable component.
diphtheria antitoxin, giving a precipitate. Sterility (2.6.1). It complies with the test far sterility.
B. Tetanus toxoid is identified by a suitable immunochemical Bacteria! endotoxins (2.6.14). The content is within the limits
method (2.7.1). The fallowing method, applicable to certain approved by the competent authority far the haemophilus
vaccines, is given as an example. The clear supernatant component of the particular product. If any components of
obtained during identification test A reacts with a suitable the vaccine prevent the determination of endotoxin, a test for
tetanus antitoxin, giving a precipitate. pyrogens is carried out as described under General provisions.
Haemophilus type b conjugate vaccine EUROPEAN PHARMACOPOEIA 9.5
ASSAY type b, covalently bound to a carrier protein. The

Diphtheria component. Carry out one of the prescribed polysaccharide, polyribosylribitol phosphate, referred to
methods for the assay of diphtheria vaccine (adsorbed) (2.7.6). as PRP, is a linear copolymer composed of repeated units
of 3-~-D-ribofuranosyl-(l ~ l)-ribitol-5-phosphate
[(C 10 H 1p 12 P)n], with a defined molecular size. The
is not less than 30 IU per single human <lose.
carrier protein, when conjugated to PRP, is capable of
Tetanus component. Carry out one of the prescribed methods inducing a T-cell-dependent B-cell immune response to the
for the assay of tetanus vaccine (adsorbed) (2.7.8). polysaccharide.
If the test is carried out in guinea-pigs, the lower confidence
limit (P = 0.95) of the estimated potency is not less than 40 IU PRODUCTION
per single human do se; if the test is carried out in mice, the GENERAL PROVISIONS
lower confidence limit (P = 0.95) of the estimated potency is
The production method shall have been shown to yield
not less than 60 IU per single human <lose.
consistently haemophilus type b conjugate vaccines of
Pertussis component. Carry out the assay of pertussis vaccine adequate safety and immunogenicity in man. The production
(whole cell) (2.7.7). of PRP and of the carrier protein are based on seed-lot systems.
The estimated potency is not less than 4.0 IU per single human The production method is validated to demonstrate that the
<lose and the lower confidence limit (P = 0.95) of the estimated product, if tested, would comply with the test for abnormal
potency is not less than 2.0 IU per single human <lose. toxicity for immunosera and vaccines for human use (2.6.9)
Poliomyelitis component and also with the test for pyrogens (2.6.8), carried out as
D-antigen con ten t. As a measure of consistency of production, follows: inject per kilogram of the rabbit's mass a quantity
determine the D-antigen content for human poliovirus of the vaccine equivalent to: 1 µg of PRP for a vaccine with
types l, 2 and 3 by a suitable immunochemical method (2.7.1) diphtheria toxoid or CRM 197 diphtheria protein as carrier;
following desorption using a reference preparation calibrated 0.1 µg of PRP for a vaccine with tetanus toxoid as carrier;
in European Pharmacopoeia Units of D-antigen. For 0.025 µg of PRP for a vaccine with OMP (meningococcal
each type, the content, expressed with reference to the group B outer membrane protein complex) as carrier.
amount of D-antigen stated on the label, is within the limits During development studies, it shall be demonstrated that
approved for the particular product. Poliomyelitis vaccine the vaccine consistently induces a T-cell-dependent B-cell
(inactivated) BRP is calibrated in European Pharmacopoeia immune response to PRP. If the manufacturing process is
Units and intended for use in the assay of D-antigen. The modified, it shall be demonstrated by appropriate in vitro
European Pharmacopoeia Unit and the International Unit are methods that the characteristic properties of the vaccine are
equivalent. not affected.
In vivo test. The vaccine complics vvith thc in vivo assay of The stability of the final lot and relevant intermediates is
poliomyelitis vaccine (inactivated) (2.7.20). evaluated using one or more indicator tests. Such tests may
LABELLING include determination of molecular size, determination of free
The label states: PRP in the conjugate and the immunogenicity test in mice.
Taking account of the results of the stability testing, release
- the minimum number of International Units of diphtheria requirements are set for these indicator tests to ensure that the
and tetanus toxoid per single human <lose; vaccine will be satisfactory at the end of the period of validity.
- the mínimum number of International Units of pertussis
BACTERIAL SEED LOTS
vaccine per single human <lose;
The seed lots of H. influenzae type b are shown to be free from
- the nominal amount of poliovirus of each type (1, 2 and 3),
contamination by methods of suitable sensitivity. These may
expressed in European Pharmacopoeia Units of D-antigen,
include inoculation into suitable media, examination of colony
per single human <lose;
morphology, microscopic examination of Gram-stained
- the type of cells used for production of the poliomyelitis smears and culture agglutination with suitable specific
component; antisera.
- the number of micrograms of PRP per single human <lose;
No complex products of animal origin are included in the
- the type and nominal amount of carrier protein per single medium used for preservation of strain viability, either for
human <lose; freeze-drying or for frozen storage.
- where applicable, that the vaccine is intended for primary
It is recommended that PRP produced by the seed lot
be characterised using nuclear magnetic resonance
reinforcing doses or for administration to adults;
spectrometry (2.2.33).
- the name and the amount of the adsorbent;
H. INFLUENZAE TYPE b POLYSACCHARIDE (PRP)
- that the vaccine must be shaken before use;
H. influenzae type b is grown in a liquid medium that <loes
- that the vaccine is not to be frozen. not contain high-molecular-mass polysaccharides; if any
ingredient of the medium contains blood-group substances,
07/2018:1219 the process shall be validated to demonstrate that after the
purification step they are no longer detectable. The bacteria!
purity of the culture is verified by methods of suitable
sensitivity. These may include inoculation into suitable media,
examination of colony morphology, microscopic examination
HAEMOPHILUS TYPE b CONJUGATE of Gram-stained smears and culture agglutination with
VACCINE suitable specific antisera. The culture may be inactivated.
PRP is separated from the culture medium and purified by
a suitable method. Volatile matter, including water, in the
Vaccinum haemophili stirpis b coniugatum purified polysaccharide is determined by a suitable method;
DEFINITION the result is used to calculate the results of certain tests with
Haemophilus type b conjugate vaccine is a liquid or reference to the dried substance, as prescribed below.
freeze-dried preparation of a polysaccharide, derived Only PRP that complies with the following requirements may
from a suitable strain of Haemophilus influenzae be used in the preparation of the conjugate.

EUROPEAN PHARMACOPOEIA 9.5 Haemophilus type b conjugate vaccine
Identification. PRP is identified by an immunochemical an appropriate detection system. An acceptable value is

method (2.7.1) or other suitable method, for example 1H established for the bulk conjugate. Each batch must be shown
nuclear magnetic resonance spectrometry (2.2.33). to comply with this limit.
Molecular-size or molecular-mass distribution. Free PRP. A number of methods are used to separate
Molecular-size or molecular-mass distribution is determined free PRP from the conjugate, including precipitation, gel
by size-exclusion chromatography (2.2.30) combined with filtration, size-exclusion, anion-exchange and hydrophobic
an appropriate detection system. Where applicable, the chromatography, ultrafiltration and ultracentrifugation. The
molecular-size distribution is also determined after chemical free PRP can then be quantified by a range of techniques,
modification of the polysaccharide. An acceptable value is including high-performance anion-exchange chromatography
established for the PRP polysaccharide. Each batch must be with pulsed amperometric detection (HPAEC-PAD) and
shown to comply with this limit. immunoassays with anti-PRP antibodies.
Ribose (2.5.31): within the limits approved by the competent Free carrier protein. Determine the content by a suitable
authority for the particular product, calculated with reference method, either directly or by deriving the content by
to the dried substance. calculation from the results of other tests. The amount is
Phosphorus (2. 5.18): within the limits approved by the within the limits approved for the particular product.
competent authority for the particular product, calculated Unreacted functional groups. No unreacted functional
with reference to the dried substance. groups are detectable in the bulk conjugate unless process
Protein (2.5.16): maximum 1.0 per cent, calculated with validation has shown that unreacted functional groups
reference to the dried substance. Use sufficient PRP to allow detectable at this stage are removed during the subsequent
detection of proteins at concentrations of 1 per cent or greater. manufacturing process (for example, owing to short half-life).
Nudeic acid (2.5.17): maximum 1.0 per cent, calculated with Residual reagents. Removal of residual reagents such as
reference to the dried substance. cyanide, EDAC (ethyldimethylaminopropylcarbodiimide)
and phenol is confirmed by suitable tests or by validation of
Bacteria} endotoxins (2.6.14): less than 10 IU per microgram the process.
of PRP.
Sterility (2.6.1). Carry out the test using for each medium
Residual reagents. Where applicable, tests are carried out 1O mL or the equivalent of 100 doses, whichever is less.
to determine residues of reagents used during inactivation
and purification. An acceptable value for each reagent is FINAL BULK VACCINE
established for the particular product and each batch of PRP An adjuvant, an antimicrobial preservative and a stabiliser
must be shown to comply with this limit. Where validation may be added to the bulk conjugate before dilution to the final
studies have demonstrated removal of a residual reagent, the concentration with a suitable diluent.
test on PRP may be omitted. Only a final bulk vaccine that complies with the following
CARRIER PROTEIN requirements may be used in the preparation of the final lot.
The production and characteristics of the carrier proteins Antimicrobial preservative. Where applicable, determine the
are described in general chapter 5.2.11. Carrier proteins amount of antimicrobial preservative by a suitable chemical or
for the production of conjugated polysaccharide vaccines for physico-chemical method. The content is not less than 85 per
human use. Only a carrier protein that complies with the cent and not greater than 115 per cent of the intended amount.
requirements of this chapter may be used in the preparation
Sterility (2.6.1). lt complies with the test for sterility, carried
of the conjugate.
out using 10 mL for each medium.
BULK CONJUGATE
FINALLOT
PRP is chemically modified to enable conjugation; it is usually
partly depolymerised either before or during this procedure. Only a final lot that is satisfactory with respect to each of
Reactive functional groups or spacers may be introduced into the following requirements and the requirements given
the appropriate carrier protein or PRP prior to conjugation. below under Identification and Tests may be released for use.
As a measure of consistency, the extent of derivatisation is Provided the test for antimicrobial preservative has been
monitored. The conjugate is obtained by the covalent binding carried out on the final bulk vaccine, it may be omitted on
of PRP and the appropriate carrier protein. Where applicable, the final lot.
unreacted but potentially reactogenic functional groups are pH (2.2.3). The pH of the vaccine, reconstituted if necessary,
made unreactive by means of capping agents; the conjugate is is within the limits approved for the particular product.
purified to remove reagents. Free PRP. A number of methods are used to separate
Only a bulk conjugate that complies with the following free PRP from the conjugate, including precipitation, gel
requirements may be used in the preparation of the final bulk filtration, size-exclusion, anion-exchange and hydrophobic
vaccine. Por each test and for each particular product, limits chromatography, ultrafiltration and ultracentrifugation. The
of acceptance are established and each batch of conjugate free PRP can then be quantified by a range of techniques,
must be shown to comply with these limits. Por a freeze-dried including HPAEC-PAD and immunoassays with anti-PRP
vaccine, sorne of the tests may be carried out on the final lot antibodies. The amount of free PRP is not greater than that
rather than on the bulk conjugate where the freeze-drying approved for the particular product.
process may affect the component being tested.
PRP. The PRP content is determined either by assay IDENTIPICATION
of ribose (2.5.31) or phosphorus (2.5.18), by an The vaccine is identified by a suitable immunochemical
immunochemical method (2.7.1) or by anion-exchange liquid method (2 .7.1) for PRP.
chromatography (2.2.29) with pulsed amperometric detection.
Protein. The protein content is determined by a suitable TESTS
chemical method (for example, 2.5.16).
PRP: mínimum 80 per cent of the amount of PRP stated on the
PRP-to-protein ratio. Determine the ratio by calculation. label. PRP is determined either by assay of ribose (2.5.31) or
Molecular-size or molecular-mass distribution. phosphorus (2.5.18), by an immunochemical method (2.7.1)
Molecular-size or molecular-mass distribution is determined or by anion-exchange liquid chromatography (2.2.29) with
by size-exclusion chromatography (2.2.30) combined with pulsed amperometric detection.
Haemophilus type b conjugate vaccine EUROPEAN PHARMACOPOEIA 9.5
Aluminium (2.5.13): maximum 1.25 mg per single human Bacteria! endotoxins (2.6.14). The content is within the
dose, if aluminium hydroxide or hydrated aluminium limits approved by the competent authority for the particular
phosphate is used as the adsorbent. product. If any components of the vaccine prevent the
Antimicrobial preservative. Where applicable, determine the determination of endotoxin, a test for pyrogens is carried out
amount of antimicrobial preservative by a suitable chemical as described under General provisions.
or physico-chemical method. The content is not less than the
minimum amount shown to be effective and not greater than LABELLING
115 per cent of the quantity stated on the label. The label states :
Water (2.5.12): maximum 3.0 per cent for freeze-dried - the number of micrograms of PRP per single human dose;
vaccines. - the type and nominal amount of carrier protein per single
Sterility (2.6.1). It complies with the test for sterility. human dose.

Vaccines for veterinary use

Foot-and-mouth disease (ruminants) vaccine Infectious chicken anaemia vaccine (live) ........................... 5598
(inactivated) .......................................................................... 5597

EUROPEAN PHARMACOPOEIA 9.5 Foot-and-mouth disease (ruminants) vaccine (inactivated)
01/2017:0063 The vaccine complies with the test if no animal shows

corrected 9.5 abnormal local or systemic reactions, or dies from causes
attributable to the vaccine.
2-4-2. lmmunogenicity. Carry out an immunogenicity test
for each strain of foot-and-mouth disease virus that may be
included in the vaccine.
FOOT-AND-MOUTH DISEASE Each test is carried out for each route and method of
administration to be recommended for vaccination, using
(RUMINANTS) VACCINE in each case cattle not less than 6 months old. The vaccine
(INACTIVATED) administered to each cattle is of minimum antigen content.
Either of the following 2 tests is suitable to demonstrate
Vaccinum aphtharum epizooticarum immunogenicity of the vaccine for cattle.
2-4-2-1. PD 50 challenge test. The potency ofthe vaccine is
inactivatum ad ruminantes expressed as the number of 50 per cent cattle protective doses
(PD 50 ) contained in the dose stated on the label. The PD 50 is
l. DEPINITION determined in cattle given primary vaccination and challenged
Poot-and-mouth disease (ruminants) vaccine (inactivated) by the inoculation of 1O 000 ID 50 of virulent bovine virus of
is a preparation containing one or more suitable strains of the same strain as that used in the preparation of the vaccine
foot-and-mouth disease virus inactivated while maintaining in the conditions described below. The vaccine virus may be
adequate immunogenic properties. This monograph applies used for challenge.
to vaccines intended for active immunisation of ruminants Use for the test not fewer than 17 cattle obtained from areas
against foot-and-mouth disease. free from foot-and-mouth disease, that have never been
vaccinated against foot-and-mouth disease and do not have
2. PRODUCTION antibodies neutralising the different strains of foot-and-mouth
2-1. PREPARA TION OF THE VACCINE disease virus. Vaccinate not fewer than 3 groups of not fewer
The vaccine virus is grown in cell cultures and then than 5 cattle per group, using a different dose of the vaccine
separated from cellular material by filtration or other suitable for each group. Administer the different doses by injecting
procedures. The harvested virus is inactivated in suitable different volumes of the vaccine and not by dilution of the
conditions and may be concentrated and purified. It is used vaccine. Maintain 2 cattle as controls. Por example, if the
for the preparation of vaccine immediately or after storage at label states that the injection of 2 mL corresponds to the
administration of 1 do se of vaccine, a 1/4 <lose of vaccine
a temperature shown to be consistent with antigen stability.
The vaccine is prepared from inactivated virus by blending would be obtained by injecting 0.5 mL, anda 1/10 <lose would
with one or more adjuvants. Por a given strain, the quantity be obtained by injecting 0.2 mL. Challenge all the cattle after
of l 46S antigen blended in each batch of vaccine is not lower 20-22 days by the intradermal route, into at least 2 sites on
than that of a batch of vaccine that has been found satisfactory the upper surface of the tongue (O.l mL per site), with a <lose
with respect to Immunogenicity. equivalent to approximately 10 000 ID 50 of a suspension of
a fully virulent virus, obtained from cattle and of the same
'1 '1 C'TTPC"T'D ¡\ 'T'TJ TJnD lTTDTTC' DDnDA ~ L1 'T'Tn1\T
""-"""· r
LJVLJLJ.L.J.\.Il..J...L..J i \J.J.'\. strain as lhal usc:<l in lhe preparation of the vaccine. Observe
.J..J.'\.VLJ .J. .L'\.\J'.L.J.1'-111.1...L\J'.L~
2-2-1. Cell cultures. The cell cultures comply with the the cattle at least daily for 8 days. In the interest of animal
requirements for cell cultures for production of veterinary welfare, individual animals may be euthanised before the end
vaccines (5.2.4). of the observation period and considered as unprotected if a
vaccinated animal shows lesions of foot-and-mouth disease
2-3. VALIDATION OF THE INACTIVATION PROCEDURE on at least 1 foot or if a control animal shows lesions of
During inactivation, the virus titre is monitored by a sensitive foot-and-mouth disease on at least 3 feet. Unprotected cattle
and reproducible technique. The inactivation procedure is show lesions at sites other than the tongue. Protected cattle
not satisfactory unless the decrease in virus titre, plotted may display lingual lesions.
logarithmically, is linear and extrapolation indicates that
The test is not valid if both control cattle do not show lesions
there is less than 1 infectious virus unit per 10 4 L of liquid
on at least 3 feet. Prom the number of protected cattle in each
preparation at the end of inactivation.
group, calculate the PD 50 content of the vaccine.
2-4. CHOICE OF VACCINE COMPOSITION The vaccine complies with the test if the potency is not less
The vaccine is shown to be satisfactory with respect to safety than that to be stated on the label; the minimum potency to
(5.2.6) and efficacy (5.2.7) for each species for which it is be stated on the label is not less than 3 PD 50 per dose for cattle.
intended. 2-4-2-2. PPG test. The following test could also be used to
The following tests for safety (section 2-4-1) and demonstrate immunogenicity of the vaccine for cattle (referred
immunogenicity (section 2-4-2) may be used during the to as the 'Percentage of protection against generalised foot
demonstration of safety and efficacy. infection' (PPG test)).
2-4-1. Safety The potency of the vaccine is expressed as the percentage
of cattle that do not show lesions on any feet. The PPG is
Carry out the test for each route and method of administration determined in cattle given primary vaccination and challenged
to be recommended for vaccination and in each category of by the inoculation of 10 000 ID 50 ofvirulent virus of the same
each species for which the vaccine is intended, using in each strain as that used in the preparation of the vaccine under
case animals of the minimum age to be recommended. Use the conditions described below. The vaccine virus may be
a representative batch of vaccine containing not less than the used for challenge. Use for the test not fewer than 18 cattle
maximum antigen content that may be expected in a batch obtained from areas free from foot-and-mouth disease, that
of vaccine. have never been vaccinated against foot-and-mouth disease
Por each test, use not fewer than 8 animals that do not have and do not have antibodies neutralising the different strains
antibodies against foot-and-mouth disease virus. Administer of foot-and-mouth disease virus. Vaccinate not fewer than
to each animal 1 dose of the vaccine. If the schedule to be 16 cattle with 1 full dose. Maintain 2 cattle as controls.
recommended requires a 2nd <lose, administer 1 dose after an Challenge ali the cattle after 20-22 days by the intradermal
interval of at least 14 days. Observe the animals at least daily route, into at least 2 si tes on the upper surface of the tongue
for at least 14 days after the last administration. (0.1 mL per site), with adose equivalent to approximately
Infectious chicken anaemia vaccine (live) EUROPEAN PHARMACOPOEIA 9.5
1O 000 ID 50 of a suspension of a fully virulent virus of the same 2-5-4-2. Vaccines for use in other ruminants. The potency of
strain as that used in the preparation of the vaccine. Observe each batch shall be demonstrated in a suitable, validated test.
the cattle at least daily for 8 days. In the interest of animal EMERGENCY USE: in situations of extreme urgency and
welfare, individual animals may be euthanised before the end subject to agreement by the competent authority, a batch of
of the observation period and considered as unprotected if a vaccine may be released before completion of the tests and the
vaccinated animal shows lesions of foot-and-mouth disease determination of potency if a test for sterility has been carried
on at least 1 foot or if a control animal shows lesions of out on the bulk inactivated antigen and all other components
foot-and-mouth disease on at least 3 feet. Unprotected cattle of the vaccine and if the determination of potency has been
show lesions at sites other than the tongue. Protected cattle carried out on a representative batch of vaccine prepared from
may display lingual lesions. the same bulk inactivated antigen. In this context, a batch is
The test is not valid if both control cattle do not show lesions not considered to be representative unless it has been prepared
on at least 3 feet. From the number of protected cattle in the with not more than the amount of antigen or antigens and
vaccinated group, calculate the percentage of protected cattle. with the same formulation as the batch to be released.
The vaccine complies with the test if the potency is not less 3. BATCH TESTS
than that to be stated on the label; the mínimum potency to 3-1. Identification. The vaccine contains the antigen or
be stated on the label is not less than 75 per cent. antigens stated under Definition.
2-5. MANUFACTURER'S TESTS 3-2. Bacteria and fungi. The vaccine and, where applicable,
2-5-1. Identification. The bulk inactivated antigen is the liquid supplied with it, comply with the test for sterility
identified by a suitable immunochemical method (2.7.1). prescribed in the monograph Vaccines for veterinary use
(0062).
2-5-2. Residual live virus. The limit of detection of the cell
cultures to be used with respect to the virus to be tested is 3-3. Potency. The vaccine complies with the requirements
established by determining the number of CCID 50 and the of the test mentioned under Immunogenicity (section 2-4-2)
1465 antigen content of a sample of live virus. The cells are when administered by a recommended mute and method.
not suitable if an amount of virus corresponding to 1 µg
of 1465 antigen has less than 106 CCID 50 • A proportion of
each batch of bulk inactivated antigen representing at least 01/2017:2038
200 doses is tested for freedom from live virus by inoculation corrected 9.5
into suitable cell cultures. A passage is made during culture
of the cells. For this purpose, the sample of the inactivated
antigen may be concentrated to allow testing of such large
samples in cell cultures, It must be shown that the selected
concentration and assay systems are not detrimental to INFECTIOUS CHICKEN ANAEMIA
detection of infectious virus within the test sample and that
the concentrated inactivated antigen <loes not interfere with VACCINE (LIVE)
virus replication or cause toxic changes. A positive control is
included in each test. Vaccinum anaemiae infectivae pulli vivum
2-5-3. Antigen content. The 1465 antigen content of each l. DEFINITION
batch of bulk inactivated antigen is determined by an in
vitro method (for example, by sucrose density gradient Infectious chicken anaemia vaccine (live) is a preparation of
centrifugation and ultraviolet spectrophotometry at 259 nm). a suitable strain of chicken anaemia virus. This monograph
applies to vaccines intended for administration to breeder
2-5-4. Batch potency test. It is not necessary to carry out chickens for active immunisation, to prevent excretion of the
the potency test (section 3-3) for each batch of vaccine virus, to prevent or reduce egg transmission and to protect
if it has been carried out using a batch of vaccine with a passively their future progeny.
mínimum antigen content. Where the test is not carried
out, an alternative validated method is used, the criteria for 2. PRODUCTION
acceptance being set with reference to a batch of vaccine 2-1. PREPARATION OF THE VACCINE
that has given satisfactory results in the test described under The vaccine virus is grown in embryonated hens' eggs or in
Potency and has been shown to be satisfactory with respect to
cell cultures.
immunogenicity in the target species.
2-2. SUBSTRATE POR VIRUS PROPAGATION
The following test may be used after a satisfactory pass level
for a given strain has been established. Once a pass level has 2-2-1. Embryonated hens' eggs. If the vaccine virus is grown
been established for a given strain, the same level of antigen in embryonated hens' eggs, they are obtained from flocks free
may be used when this strain is formulated in combination from specified pathogens (SPF) (5.2.2).
with any other antigen provided that the formulation of the 2-2-2. Cell cultures. If the vaccine virus is grown in cell
vaccine differs only in the strains included. cultures, they comply with the requirements for cell cultures
for production of veterinary vaccines (5.2.4).
2-5-4-1. Vaccines for use in cattle. Use cattle of the mínimum
age recommended for vaccination obtained from areas free 2-3. SEED LOTS
from foot-and-mouth disease, that have never been vaccinated 2-3-1. Extraneous agents. The master seed lot complies with
against foot-and-mouth disease and do not have antibodies the test for extraneous agents in seed lots (2.6.24). In these
neutralising the different strains of foot-and-mouth disease tests on the master seed lot, the organisms used are not more
virus. Vaccinate not fewer than 5 cattle by a recommended than 5 passages from the master seed lot at the start of the tests.
mute. Use a suitable <lose of the vaccine for each animal.
After a defined period, not greater than 28 days following 2-4. CHOICE OF VACCINE VIRUS
vaccination, draw a blood sample and determine individually The vaccine virus is shown to be satisfactory with respect to
in each serum the level of antibodies against each strain used safety (5.2.6) and efficacy (5.2.7) for the chickens for which
in the preparation of the vaccine by a validated technique (e.g. it is intended.
sero-neutralisation test, ELI5A). The vaccine complies with The following tests for safety (section 2-4-1), increase in
the test if the geometric mean of the antibody titre in cattle is virulence (section 2-4-2) and immunogenicity (section 2-4-3)
not significantly lower than the pass level. may be used during the demonstration of safety and efficacy.

EUROPEAN PHARMACOPOEIA 9.5 Infectious chicken anaemia vaccine (live)
2-4-1. Safety. Carry out the test for each route and method observed. If virus is not recovered after an initial passage in
of administration to be recommended for vaccination in 5 chickens and a subsequent repeat passage in 10 chickens, the
chickens not older than the minimum age to be recommended vaccine virus also complies with the test.
for vaccination and from an SPF flock (5.2.2). Use vaccine 2-4-3. Immunogenicity. A test is carried out for each route
virus at the least attenuated passage level that will be present and method of administration to be recommended for
in a batch of the vaccine. vaccination using chickens not older than the minimum age
2-4-1-1. General safety. For each test, use not fewer than to be recommended for vaccination and from an SPF flock
8 chickens. Administer to each chicken a quantity of the (5.2.2). The test for prevention of virus excretion is intended
vaccine virus equivalent to not less than 10 times the to demonstrate reduction of egg transmission through
maximum virus titre likely to be contained in 1 <lose of the viraemia and virus excretion in the faeces. The quantity of the
vaccine. 14 days after vaccination, collect blood samples vaccine virus to be administered to each chicken is not greater
from half of the chickens and determine the haematocrit than the minimum virus titre to be stated on the label and
value. Euthanise these chickens and carry out post-mortem the virus is at the most attenuated passage level that will be
examination. Note any pathological changes attributable to present in a batch of vaccine.
chicken anaemia virus, such as thymic atrophy and specific 2-4-3-1. Passive immunisation of chickens. Vaccinate
bone-marrow lesions. Observe the remaining chickens at least according to the schedule to be recommended not fewer than
daily, for at least 21 days after vaccination. 10 breeder chickens not older than the mínimum age to be
The test is not valid if non-specific mortality occurs. recommended for vaccination and from an SPF flock (5.2.2);
The vaccine virus complies with the test if during the keep not fewer than 1O unvaccinated breeder chickens of the
observation period no chicken shows abnormal signs of same origin and from an SPF flock (5.2.2) as controls that have
disease or dies from causes attributable to the vaccine virus. no contact with the vaccinated chickens. At a suitable time,
after excretion of vaccine virus has ceased, collect fertilised
2-4-1-2. Safety for young chickens. Use not fewer than twenty eggs from the vaccinated and control breeder chickens and
1-day-old chickens from an SPF flock (5.2.2). Administer to incubate them. Challenge at least 30 1-day-old chickens from
each chicken by the oculonasal route a quantity of the vaccine each of the vaccinated and control groups by intramuscular
virus equivalent to not less than the maximum titre likely to administration of a sufficient quantity of virulent chicken
be contained in 1 <lose of the vaccine. Observe the chickens anaemia virus. Observe the chickens at least daily for 14 days
at least daily. Record the incidence of any signs attributable after challenge. Record the deaths and the surviving chickens
to the vaccine virus, such as depression, and any deaths. that show signs of disease. At the end of the observation
14 days after vaccination, collect blood samples from half of period determine the haematocrit value of each surviving
the chickens and determine the haematocrit value. Euthanise chicken. Euthanise these chickens and carry out post-mortem
these chickens and carry out post-mortem examination. Note examination. Note any pathological signs attributable to
any pathological changes attributable to chicken anaemia chicken anaemia virus, such as thymic atrophy and specific
virus, such as thymic atrophy and specific bone marrow bone-marrow lesions.
lesions. Observe the remaining chickens at least daily, for at
least 21 days after vaccination. Assess the extent to which the The test is not valid if:
vaccine strain is pathogenic for 1-day-old susceptible chickens - the laying rate in the vaccinated and control breeder
from the results of the clinical observations and mortality rates chickens is significantly different;
and the proportion of chickens examined at 14 days that show
- during the observation period after challenge fewer than
anaemia (haematocrit value less than 27 per cent) and signs
90 per cent of the chickens of the control breeder chickens
of infectious chicken anaemia on post-mortem examination.
die or show severe signs of infectious chicken anaemia,
The results are used to formulate the label statement on safety
including haematocrit value under 27 per cent, and/or
for young chickens.
notable macroscopic lesions of the bone marrow and
2-4-2. Increase in virulence. Carry out the test according thymus;
to general chapter 5.2.6 using 1-day-old chickens from an
- and/or during the period between vaccination and egg
SPF flock (5.2.2). If the properties of the vaccine virus allow
collection more than 10 per cent of vaccinated or control
sequential passage through 5 groups via natural spreading,
breeder chickens show notable signs of disease or die from
this method may be used, otherwise passage as described
causes not attributable to the vaccine.
below is carried out.
Administer to each chicken of the 1st group by the The vaccine complies with the test if during the observation
period after challenge not fewer than 90 per cent of the
intramuscular route a quantity of the vaccine virus that will
chickens of the vaccinated breeder chickens survive and show
allow recovery of virus for the passages described below.
no notable signs of disease and/ or macroscopic lesions of the
Prepare 7-9 days after administration a suspension from the
bone marrow and thymus.
liver of each chicken and pool these samples. Depending
on the tropism of the virus, other tissues such as spleen or 2-4-3-2. Prevention of virus excretion. Vaccinate according to
bone marrow may be used. Administer 0.1 mL of the pooled the schedule to be recommended not fewer than 1O chickens
samples by the intramuscular route to each chicken of the not older than the minimum age to be recommended for
next group. Carry out this passage operation not fewer than vaccination and from an SPF flock (5.2.2). Maintain not fewer
4 times; verify the presence of the virus at each passage. If than 1O chickens of the same age and origin as controls that
the virus is not found at a passage level, repeat the passage by have no contact with the vaccinated chickens. At a suitable
administration to a group of 10 chickens. time after excretion of vaccine virus has ceased, challenge all
If the 5th group of chickens shows no evidence of an increase the chickens by intramuscular administration of a sufficient
in virulence indicative of reversion during the observation quantity of virulent chicken anaemia virus. Collect blood
period, further testing is not required. Otherwise, carry out an samples from the chickens on days 3, 5 and 7 after challenge
additional safety test and compare the clinical signs and any and faecal samples from the chickens on days 7, 14 and 21
relevant parameters in a group of at least 1O chickens receiving after challenge and carry out a test for presence of virus to
the material used for the 1st passage and another similar group determine whether or not the chickens are viraemic and are
receiving the virus at the final passage level. excreting the virus.
The vaccine virus complies with the test if no indication The test is not valid if:
of increased virulence of the virus at the final passage - fewer than 70 per cent of the control chickens are viraemic
level compared with the material used for the 1st passage is and excrete the virus at one or more times of sampling;
Infectious chicken anaemia vaccine (live) EUROPEAN PHARMACOPOEIA 9.5
- and/or during the period between vaccination and contain pathogenic micro-organisms and contains not more
challenge more than 1O per cent of control or vaccinated than 1 non-pathogenic micro-organism per <lose.
chickens show abnormal clinical signs or die from causes Any diluent supplied for reconstitution of the vaccine complies
not attributable to the vaccine. with the test for sterility prescribed in the monograph Vaccines
The vaccine complies with the test if not fewer than 90 per for veterinary use (0062).
cent of the vaccinated chickens do not develop viraemia or 3-3. Mycoplasmas. The vaccine complies with the test for
excrete the virus. mycoplasmas (2.6.7).
3-4. Extraneous agents. The vaccine complies with the tests
3. BATCH TESTING for extraneous agents in batches of finished product (2.6.25).
3-1. Identification. The vaccine, diluted if necessary and 3-5. Virus titre. Titrate the vaccine virus by inoculation into
mixed with a monospecific chicken anaemia virus antiserum, suitable cell cultures (5.2.4) or eggs from an SPF flock (5.2.2).
no longer infects susceptible cell cultures or eggs from an SPF The vaccine complies with the test if 1 <lose contains not less
flock (5.2.2) into which it is inoculated. than the minimum virus titre stated on the label.
3-2. Bacteria and fungi. Vaccines intended for administration 3-6. Potency. The vaccine complies with the requirements of
the tests prescribed under Immunogenicity (sections 2-4-3-1
by injection comply with the test for sterility prescribed in the
monograph Vaccines for veterinary use (0062). and 2-4-3-2) when administered by a recommended route
and method. It is not necessary to carry out the potency test
Frozen or freeze-dried vaccines produced in embryonated for each batch of the vaccine if it has been carried out on a
eggs and not intended for administration by injection representative batch using a vaccinating <lose containing not
either comply with the test for sterility prescribed in the more than the minimum virus titre stated on the label.
monograph Vaccines for veterinary use (0062) or with the
following test: carry out a quantitative test for bacteria! 4. LABELLING
and fungal contamination; carry out identification tests for The label states to which extent the vaccine virus causes
micro-organisms detected in the vaccine; the vaccine does not disease if it spreads to susceptible young chickens.

Sutures for human use

Sutures, sterile non-absorbable ............................................. 5603

EUROPEAN PHARMACOPOEIA 9.5 Sutures, sterile non-absorbable
07 /2018:0324 Stainless steel sutures consist of smooth, cylindrical

monofilaments or twisted filaments or braided filaments.
Poly(vinylidene difluoride) (PVDF) (Filum poly(vinylideni
difluoridum))
Sterile PVDF suture is obtained by drawing through a suitable
SUTURES, STERILE NON- die a synthetic plastic material formed by polymerisation
ABSORBABLE of 1, 1-difluoroethylene. It consists of smooth, cylindrical
monofilaments.
Fila non resorbilia sterilia IDENTIFICATION
DEFINITION Synthetic and natural materials may be identified by infrared
Sterile non-absorbable sutures are sutures that, when spectrophotometry (2.2.24) using attenuated total reflection
introduced into a living organism, are not metabolised by (ATR) or by differential scanning calorimetry. Additives and
that organism. Sterile non-absorbable sutures vary in origin, coatings of materials may lead to additional peaks. Natural
which may be animal, vegetable, metallic or synthetic. They materials may also be identified by microscopic examination
occur as cylindrical monofilaments or as multifilament sutures of the morphology of the fibres.
consisting of elementary fibres that are assembled by twisting, Identification of silk
cabling or braiding; they may be sheathed; they may be A. Dissect the end of a suture, using a needle or fine tweezers,
treated to render them non-capillary; they may be coloured. to isolate a few individual fibres. The fibres are sometimes
Appropriate harmonised standards may be considered when marked with very fine longitudinal striations parallel to
assessing compliance with respect to origin and processing of the axis of the suture. Examined under a microscope, a
raw materials and with respect to biocompatibility. cross-section is more or less triangular to semi-circular,
Sterile non-absorbable surgical sutures serve to approximate with rounded edges and without a lumen.
tissue during the healing period and provide continuing B. Infrared absorption spectrophotometry (2.2.24). Examine
wound support. by attenuated total reflection (ATR).
The following materials are frequently used, as well as blends Absorption maxima and intensities: 3280 ± 5 cm- 1
thereof, as blends of synthetic materials are common. (strong); 2923 ± 15 cm- 1 (medium); 1622 ± 15 cm- 1
Silk (Filum bombycis) (strong); 1512 ± 5 cm- 1 (strong); 1444 ± 7 cm- 1 (medium);
1226 ± 10 cm- 1 (medium).
Sterile braided silk suture is obtained by braiding a number of
threads, according to the diameter required, of degummed silk Identification of linen
obtained from the cocoons of the silkworm Bombyx morí L. A. Dissect the end of a suture, using a needle or fine tweezers,
Linen (Filum lini) to isolate a few individual fibres. Examined under a
Sterile linen thread consists of the pericyclic fibres of the stem microscope, the fibres are seen to be 12-31 µm wide and,
of Linum usitatissimum L. The elementary fibres, 2.5-5 cm along the greater part of their length, have thick walls,
long, are assembled in bundles 30-80 cm long and spun into sometimes marked with fine longitudinal striations, and
cunlinuous lengths of suitable diameter. a narrow lumen. The fibres 2:raduallv narrow to a lonQ:,
fine point. Sometimes there ~re unil~teral swellings with
Poly(ethylene terephthalate) (Filum ethyleni transverse lines.
polyterephthalici)
B. Infrared absorption spectrophotometry (2.2.24). Examine
Sterile poly(ethylene terephthalate) suture is obtained by by attenuated total reflection (ATR).
drawing poly(ethylene terephthalate) through a suitable die.
The suture is prepared by braiding very fine filaments in Absorption maxima and intensities: 3326 ± 10 cm- 1
suitable numbers, depending on the gauge required. (medium); 2911 ± 12 cm- 1 (medium); 1645 ± 10 cm- 1
(weak); 1426 ± 5 cm- 1 (medium); 1315 ± 3 cm- 1 (medium);
Polyamide 6 (Filum polyamidicum-6) 1154 ± 7 cm- 1 (medium); 1104 ± 5 cm- 1 (medium);
Sterile polyamide 6 suture is obtained by drawing through 1050 ± 5 cm- 1 (strong); 1026 ± 7 cm- 1 (strong).
a suitable die a synthetic plastic material formed by the Identification of poly( ethylene terephthalate)
polymerisation of E-caprolactam. It consists of smooth,
cylindrical monofilaments or braided filaments, or lightly It is attacked by strongly alkaline solutions. It is incompatible
twisted sutures sheathed with the same material. with phenols.
Polyamide 6/6 (Filum polyamidicum-6/6) Infrared absorption spectrophotometry (2.2.24). Examine by
attenuated total reflection (ATR).
Sterile polyamide 6/6 suture is obtained by drawing through
a suitable die a synthetic plastic material formed by the Absorption maxima and intensities: 1712.5 ± 5 cm- 1
polycondensation of hexamethylenediamine and adipic acid. (strong); 1408 ± 5 cm- 1 (medium); 1338 ± 5 cm- 1 (medium);
It consists of smooth, cylindrical monofilaments or braided 1243 ± 12 cm- 1 (strong); 1093 ± 5 cm- 1 (strong); 1017 ± 5 cm- 1
filaments, or lightly twisted sutures sheathed with the same (medium); 872 ± 5 cm- 1 (medium); 722 ± 2 cm- 1 (strong).
material. Identification of polyamide 6, polyamide 6/6 and blends
Polypropylene (Filum polypropylenicum) thereof
Polypropylene suture is obtained by drawing polypropylene They are not attacked by dilute alkaline solutions (for example
through a suitable die. It consists of smooth cylindrical a 100 g/L solution of sodium hydroxide R) but are attacked by
mono-filaments. dilute mineral acids (for example a 20 g/L solution of sulfuric
acid R) and by hot glacial acetic acid R.
Monofilament and multifilament stainless steel (Filum
aciei irrubiginibilis monofilamentum/multifilamentum) Infrared absorption spectrophotometry (2.2.24). Examine by
Sterile stainless steel sutures have a chemical composition attenuated total reflection (ATR).
as specified in ISO 5832-1 - Metallic Materials for surgical Absorption maxima and intensities: 3296 ± 10 cm- 1 (medium);
implants - Part 1 : Specification for wrought stainless steel, 2930 ± 15 cm- 1 (medium); 2862 ± 15 cm- 1 (medium);
and comply with ISO 10334 - Implants for surgery - Malleable 1635 ± 5 cm- 1 (strong);1539 ± 12 cm- 1 (strong); 1462 ± 10 cm- 1
wires for use as sutures and other surgical applications. (medium); 1262 ± 12 cm- 1 (medium).
Sutures, sterile non-absorbable EUROPEAN PHARMACOPOEIA 9.5
Identification of polypropylene It is essential for the effectiveness and the performance

Polypropylene is soluble in decahydronaphthalene, in characteristics during use and during the functional lifetime
1-chloronaphthalene and in trichloroethylene. It is insoluble of these sutures that the following physical properties are
in ethanol (96 per cent) and in cyclohexanone. specified: consistent diameter, sufficient initial strength and
Infrared absorption spectrophotometry (2.2.24). Examine by firm needle attachment.
attenuated total reflection (ATR). The requirements below have been established, taking into
Absorption maxima and intensities: 2950 ± 5 cm- 1 (strong); account stresses which occur during normal conditions of
2916 ± 5 cm- 1 (strong); 2870 ± 5 cm- 1 (medium); 2838 ± 5 cm- 1 use. These requirements can be used to demonstrate that
(medium); 1456 ± 5 cm- 1 (medium); 1376 ± 5 cm- 1 (medium). individual production batches of these sutures are suitable for
Identification of stainless steel wound closure in accordance with usual surgical techniques.
Stainless steel sutures are identified by confirming that the
composition is in accordance with ISO 5832 Part l. TESTS
Identification of poly(vinylidene difluoride) Remove the sutures from the sachet and measure promptly and
It is soluble in warm dimethylformamide. It is insoluble in in succession the length, diameter and minimum load.
anhydrous ethanol, in hot and cold isopropyl alcohol, in ethyl
acetate and in tetrachlorethylene. If linen is tested the sutures are conditioned as follows: if
Infrared absorption spectrophotometry (2.2.24). Examine by stored in the dry state, expose to an atmosphere with a relative
attenuated total reflection (ATR). humidity of 65 ± 5 per cent at 20 ± 2 ºC for 4 h immediately
before measuring the diameter and for the determination
Absorption maxima and intensities: 1399 ± 5 cm- 1 (medium);
of minimum breaking load, immerse in water R at room
1275 ± 2 cm- 1 (medium); 1165 ± 10 cm- 1 (strong); 1070 ± 5 cm- 1
temperature for 30 min immediately befare carrying out the
(medium); 873 ± 3 cm- 1 (strong); 838 ± 2 cm- 1 (strong).
test.
PRODUCTION Length. Measure the length without applying more tension
The appropriate harmonised standards apply with respect to than is necessary to keep them straight. The length of the
appropriate validated methods of sterilisation, environmental suture is not less than 95 per cent of the length stated on the
control during manufacturing, labelling and packaging. label and does not exceed 400 cm.
Table 0324.-1. - Diameters and minimum breaking loads
Diameter Mínimum breaking load
(millimetres) (newtons)
A B Ali other non-absorbable
Gauge Linen thread Stainless steel
----- strands
number
min. max. min. max. e D e D e D
o.os o.oos 0.009 0.003 0.012 0.01
0.1 0.010 0.019 O.OOS 0.02S 0.03
O.lS O.OlS 0.019 0.012 0.02S 0.06 0.01
0.2 0.020 0.029 O.OlS 0.03S 0.1
0.3 0.030 0.039 0.02S 0.04S 0.3S 0.06
0.4 0.040 0.049 0.03S 0.060 0.60 O.lS 1.1
O.S O.OSO 0.069 0.04S 0.08S 1.0 0.3S 1.6
0.7 0.070 0.099 0.060 0.12S 1.0 0.3 l.S 0.60 2.7
0.100 0.149 0.08S 0.17S 2.S 0.6 3.0 1.0 S.3 4.0
l.S O.lSO 0.199 0.12S 0.22S s.o 1.0 S.O l.S 8.0 6.0
2 0.200 0.249 0.17S 0.27S 8.0 2.S 9.0 3.0 13.3 10.0
2.S 0.2SO 0.299 0.22S 0.32S 9.0 s.o 13.0 s.o lS.S 11.6
0.300 0.349 0.27S 0.37S 11.0 8.0 lS.O 9.0 17.7 13.3
3.S 0.3SO 0.399 0.32S 0.4SO lS.O 9.0 22.0 13.0 33.4 2S.O
4 0.400 0.499 0.37S o.sso 18,0 ll.O 27.0 lS.O 46.7 3S.O
s o.soo O.S99 0.4SO 0.6SO 26.0 lS.O 3S.O 22.0 S7.9 43.4
6 0.600 0.699 o,.sso 0.7SO 37.0 18.0 so.o 27.0 89.4 67.0
7 0.700 0.799 0.6SO 0.8SO SO.O 26.0 62.0 3S.O 111.8 83.9
8 0.800 0.899 0.7SO 0.9SO 6S.O 37.0 73.0 so.o 133.4 100.1
9 0.900 0.999 0.8SO l.OSO 1S6.0 117.0
10 1.000 1.099 0.9SO l.lSO 178.S 133.9
Diameter. Unless otherwise prescribed, measure the diameter of 100 ± 10 g to the suture being tested. When making the
by the following method using 5 sutures. Use a suitable measurements, lower the pressor foot slowly to avoid crushing
mechanical instrument capable of measuring with an accuracy the suture. Measure the diameter at intervals of 30 cm over
of at least 0.002 mm and having a circular pressor foot the whole length of the suture. For a suture less than 90 cm in
10-15 mm in diameter. The pressor foot and the moving length, measure at 3 points approximately evenly spaced along
parts attached to it are weighted so as to apply a total load the suture. During the measurement submit monofilament

EUROPEAN PHARMACOPOEIA 9.5 Sutures, sterile non-absorbable
sutures to a tension not greater than that required to keep given in Table 0324.-2 for the gauge number concerned. If
them straight. Submit multifilament sutures to a tension not not more than 1 individual value fails to meet the individual
greater than one-fifth of the minimum breaking load shown in requirement, repeat the test on an additional 1O sutures. The
column C of Table 0324.-1 appropriate to the gauge number attachment complies with the test if non e of these 1O values is
and type of material concerned or 1O N whichever is less. less than the individual value in Table 0324.-2 for the gauge
Stainless steel sutures do not require tension to be applied number concerned.
during the measurement of diameter. Por multifilament
sutures of gauge number above 1.5 make 2 measurements Table 0324.-2. - Mínimum strengths of needle attachment
at each point, the second measurement being made after Gauge number Mean value Individual value
rotating the suture through 90º. The diameter of that point (newtons) (newtons)
is the average of the 2 measurements. The average of the 0.4 0.50 0.25
measurements carried out on the sutures being tested and 0.5 0.80 0.40
not less than two-thirds of the measurements taken on each
suture are within the limits given in the column under A in 0.7 1.7 0.80
Table 0324.-1 for the gauge number concerned. None of the 2.3 1.1
measurements are outside the limits given in the columns
under B in Table 0324.-1 for the gauge number concerned. 1.5 4.5 2.3
Minimum breaking load. Unless otherwise prescribed, 2 6.8 3.4

determine the minimum breaking load by the following
2.5 9.0 4.5
method using sutures in the condition in which they are
presented. The mínimum breaking load is determined over 3 ll.O 4.5
a simple knot formed by placing one end of a suture held
3.5 15.0 4.5
in the right hand over the other end held in the left hand,
passing one end over the suture and through the loop so 4 18.0 6.0
formed (see Figure 0324.-1) and pulling the knot tight. Por 5 18.0 7.0
stainless steel sutures gauges 3.5 and above, the minimum
breaking load is determined on a straight pull. Carry out the 6 25.0 12.5
test on 5 sutures. Submit sutures of length greater than 75 cm 7 25.0 12.5
to 2 measurements and shorter sutures to 1 measurement.
Determine the breaking load using a suitable tensilometer. 8 50.0 25
The apparatus has 2 clamps for holding the suture, 1 of which
9 50.0 25
is mobile and is driven ata constant rate of 30 cm/min.
The clamps are designed so that the suture being tested 10 75.0 37.5
can be attached without any possibility of slipping. At the
beginning of the test the length of suture between the clamps Extractable colour. Sutures that are dyed and intended to
is 12.5-20 cm and the knot is midway between the clamps. remain so during use comply with the test for extractable
Set the mobile clamp in motion and note the force required colour. Place 0.25 g of the suture to be examined in a conical
to break the suture. If the suture breaks in a clamp or within flask, add 25.0 mL of water R and cover the mouth of the flask
1 cm of it, the result is discarded and the test repeated on with a short-stemmed funnel. Boíl for 15 min. cool and adiust
another suture. The average of all the results, excluding those to the original volume with water R. Dependi~g on the col~ur
legitimately discarded, is equal to or greater than the value of the suture, prepare the appropriate reference solution as
given in column C in Table 0324.-1 and no value is less than described in Table 0324.-3 using the primary colour solutions
that given in column D for the gauge number and type of (2.2.2).
material concerned.
Table 0324.-3. - Colour reference solutions
Colour of Composition of reference solution
strand (parts by volume)
Red Yellow Blue
primary primary primary Water R
solution solution solution
Yellow-brown 0.2 1.2 8.6
Pink-red 1.0 9.0
Green-blue 2.0 8.0
Violet 1.6 8.4
The test solution is not more intensely coloured than the

appropriate reference solution.
Monomer and oligomers: maximum 2 per cent for the
Figure 0324.-1 . - Simple knot polyamide 6 suture.
N eedle attachment. If the sutures are supplied with an In a continuous-extraction apparatus, treat 1.00 g with 30 mL
eyeless needle attached that is not stated to be detachable, of methanol R at a rate of at least 3 extractions per hour for 7 h.
they comply with the test for needle attachment. Carry out Evaporate the extract to dryness, dry the residue at 110 ºC for
the test on 5 sutures. Use a suitable tensilometer, such as that 1O min, allow to cool in a desiccator and weigh. The residue
described for the determination of the minimum breaking weighs a maximum of 20 mg.
load. Fix the needle and suture (without knot) in the clamps
of the apparatus in such a way that the swaged part of the STORAGE (PACKAGING)
needle is completely free of the clamp and in line with the Sterile non-absorbable sutures are presented in a suitable
direction of pull on the suture. Set the mobile clamp in motion sachet that maintains sterility and allows the withdrawal and
and note the force required to break the suture or to detach use of a suture in aseptic conditions. They may be stored dry
it from the needle. The average of the 5 determinations and or in a preserving liquid to which an antimicrobial agent but
all individual values are not less than the respective values no antibiotic may be added.
Sutures, sterile non-absorbable EUROPEAN PHARMACOPOEIA 9.5
Sterile non-absorbable sutures are intended to be used only on The details strictly necessary for the user to identify the
the occasion when the sachet is first opened. product properly are indicated on or in each sachet (primary
packaging) and on the protective cover (box) and include at
Sutures in their individual sachets (primary packaging) are least:
kept in a protective cover (box) which maintains the physical
and mechanical properties until the time of use. - gauge number;
- length, in centimetres or metres;
The application of appropriate harmonised standards for - if appropriate, that the needle is detachable;
packaging of medical devices shall be considered in addition.
- name of the product;
- intended use (surgical suture, non-absorbable);
LABELLING - if appropriate, that the suture is coloured;
Reference may be made to the appropriate harmonised - if appropriate, the structure (braided, monofilament,
standards for the labellíng of medical devices. sheathed).

Sutures for veterinary use

Polyamide 6 suture, sterile, in distributor for veterinary use Poly(ethylene terephthalate) suture, sterile, in distributor for
................................................................................................ 5609 veterinary use ........................................................................ 5609
Polyamide 6/6 suture, sterile, in distributor for veterinary
use ........................................................................................... 5609
5608 See the ínformatíon section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.5 Poly( ethylene terephthalate) suture, sterile, in distributor (vet.)
m
-
07 /2018:0609 07 /2018:0610
POLYAMIDE 6/6 SUTURE, STERILE, IN

POLYAMIDE 6 SUTURE, STERILE, IN DISTRIBUTOR FOR VETERINARY USE
DISTRIBUTOR FOR VETERINARY USE
Filum polyamidicum-6/6 sterile in fusa ad
Filum palyamidicum-6 sterile in fusa usum veterinarium
ad usum veterinarium DEFINITION
Sterile polyamide 6/6 suture in distributor for veterinary
use is obtained by drawing through a suitable die a
DEFINITION synthetic plastic material formed by the polycondensation
Sterile polyamide 6 suture in distributor for veterinary use is of hexamethylene-diamine and adipic acid. It consists of
obtained by drawing through a suitable die a synthetic plastic smooth, cylindrical monofilaments or braided filaments, or
lightly twisted strands sheathed with the same material. It may
material formed by the polymerisation of E-caprolactam. It
consists of smooth, cylindrical monofilaments or braided be coloured with authorised colouring matter or pigments
filaments, or lightly twisted strands sheathed with the same authorised by the competent authority. The suture is sterilised.
material. It may be coloured with colouring matter authorised Blends of polyamide 6/6 and polyamide 6 are common.
by the competent authority. The suture is sterilised.
CHARACTERS
Blends of polyamide 6 and polyamide 6/6 are common. It is not attacked by dilute alkaline solutions (for example
a 100 g/L solution of sodium hydroxide) but is attacked by
dilute mineral acids (for example 20 g/L sulfuric acid) and by
CHARACTERS hot glacial acetic acid.
lt is not attacked by dilute alkaline solutions (for exampleIDENTIFICATION
a 100 g/L solution of sodium hydroxide) but is attacked by Infrared absorption spectrophotometry (2.2.24). Examine by
dilute mineral acids (for example 20 g/L sulfuric acid) and by
attenuated total reflection (ATR).
hot glacial acetic acid.
Absorption maxima and intensities: 3296 ± 10 cm- 1 (medium);
2930 ± 15 cm- 1 (medium); 2862 ± 15 cm- 1 (medium);
IDENTIFICATION 1635 ± 5 cm- 1 (strong); 1539 ± 12 cm- 1 (strong);
1462 ± 10 cm- 1 (medium); 1262 ± 12 cm- 1 (medium).
Infrared absorption spectrophotometry (2.2.24). Examine by Additives and coatings of materials might lead to additional
attenuated total reflection (ATR). peaks.
Absorption maxima and intensities: 3296 ± 10 cm-i (medium); TESTS
1635 ± 5 cm- 1 (strong); 1539 ± 12 cm- 1 (strong); It complies with the tests prescribed in the monograph Strands,
1462 ± 10 cm- 1 (medium); 1262 ± 12 cm- 1 (medium). sterile non-absorbable, in distributor for veterinary use (0605).
Additives and coatings of materials might lead to additional STORAGE

peaks. See the monograph Strands, sterile non-absorbable, in
dístríbutor for veterínary use (0605).
TESTS LABELLING
See the monograph Strands, sterile non-absorbable, in
It complies with the tests prescribed in the monograph distributor for veterinary use (0605).
Strands, sterile non-absorbable, in dístríbutor for veterínary
The label states whether the suture is braided, monofilament
use (0605) and with the following test.
or sheathed.
Monomer and oligomers: maximum 2 per cent.
In a continuous-extraction apparatus, treat 1.00 g with 30 mL . . . 07/2018:0607
of methanol R at a rate of at least 3 extractions per hour for 7 h.
Evaporate the extract to dryness, dry the residue at 110 ºC for
1O min, allow to cool in a desiccator and weigh. The residue
weighs a maximum of 20 mg.
POLY(ETHYLENE TEREPHTHALATE)
STORAGE SUTURE, STERILE, IN DISTRIBUTOR
FOR VETERINARY USE
distributor for veterinary use (0605).
Filum ethyleni palyterephthalici sterile in
LABELLING
fusa ad usum veterinarium
DEFINITION
distributor for veterinary use (0605). Sterile poly(ethylene terephthalate) suture in distributor
for veterinary use is obtained by drawing poly( ethylene
The label states whether the suture is braided, monofilament terephthalate) through a suitable die. The suture is prepared
or sheathed. by braiding very fine filaments in suitable numbers, depending
Poly( ethylene terephthalate) suture, sterile, in distributor (vet.) EUROPEAN PHARMACOPOEIA 9.5
on the gauge required. It may be whitish in colour, or may Additives and coatings of materials might lead to additional
be coloured with authorised colouring matter or pigments peaks.
authorised by the competent authority. The suture is sterilised.
TESTS
CHARACTERS
It is attacked by strongly alkaline solutions and is incompatible It complies with the tests prescribed in the monograph Strands,
with phenols. sterile non-absorbable, in distributor for veterinary use (0605).
IDENTIFICATION STORAGE
Infrared absorption spectrophotometry (2.2.24). Examine by
attenuated total reflection (ATR). See the monograph Strands, sterile non-absorbable, in
distributor for veterinary use (0605).
Absorption maxima and intensities: 1712.5 ± 5 cm- 1 (strong);
1243 ± 12 cm- 1 (strong); 1093 ± 5 cm- 1 (strong); LABELLING
1017 ± 5 cm- 1 (medium); 872 ± 5 cm- 1 (medium); See the monograph Strands, sterile non-absorbable, in
722 ± 2 cm- 1 (strong). distributor for veterinary use (0605).

Herbal drugs and herbal drug

preparations
Dios corea nipponica rhizome ............................................... 5613 Lavender oil.. ........................................................................... 5615
Lavender flower....................................................................... 5614 Spike lavender oil. ................................................................... 5617

EUROPEAN PHARMACOPOEIA 9.5 Dioscorea nipponica rhizome
04/2017:2890
corrected 9.5
DIOSCOREA NIPPONICA RHIZOME
Dioscoreae nipponicae rhizoma

DEFINITION
Dried, whole or fragmented, scraped rhizome of Dioscorea
nipponica Makino, with roots removed.
Content: mínimum 1.0 per cent of diosgenin (C 27 H 42 0 3 ;
Mr 414.6) (dried drug).
IDENTIFICATION
A. Whole drug. The rhizome is subcylindrical, slightly curved,
up to 15-20 cm long and 1.0-1.5 cm in diameter. The outer
surface is yellowish-white or brownish-yellow, irregularly
and longitudinally furrowed, bearing spinous remains of
roots and protuberant stem scars.
Fragmented drug. Transverse slices of the rhizome,
oblique or more or less longitudinal, circular, oval or
elongated, whole or broken, up to 0.7 cm thick. The outer
surface is light brown or yellowish-brown, irregularly and Figure 2890.-1. - Illustration for identification test B of
longitudinally furrowed; spinous remains of roots and powdered herbal drug of Dioscorea nipponica rhizome
protuberant stem scars may be visible. A transverse section
Results: see below the sequence of zones present in the
is yellowish-white, can easily be imprinted with one's
chromatograms obtained with the reference solution and
nail due to an abundance of starch, and shows numerous
the test solution. Furthermore, other faint zones may
light yellowish-brown pits corresponding to the vascular
be present in the chromatogram obtained with the test
bundles.
solution.
B. Microscopic examination (2.8.23). The powder is whitish
Top of the olate
or light yeHow. Examine under a microscope using chloral &
hydrate solution R. The powder shows the following 3 olive-green zones

diagnostic characters (Figure 2890.-1): vessels up to 50 µm
--- ---
in diameter with numerous fine, dense and elliptical
pits [G] ; numerous fragments of parenchyma with Aescin: a violet zone
polyhedral or ovoid cells with slightly thickened and pitted
A green zone
walls (transverse section [B], longitudinal section [C]);
cells containing raphides of calcium oxalate up to 11 O µm A faint to strong green zone
long [D]; free needles of calcium oxalate [A]; rare cork
Glucose: a yellow or brown zone A yellowish-brown zone
fragments with polyhedral cells (surface view [F]). Examine
under a microscope using a 50 per cent V/V solution of A faint brown zone
glycerol R. The powder shows extremely abundant starch
--- ---
granules, simple, irregular, ovoid, oblong, sub-triangular,
with a maximum dimension of up to 30 µm; the hilum is A yellow zone
usually eccentric and slit-shaped [E].
Reference soJution Test solution
C. Thin-layer chromatography (2.2.27).
Test solution. To 0.5 g of the powdered herbal drug (355) TESTS
(2.9.12) add 5 mL of methanol R. Sonicate for 10 min. Loss on drying (2.2.32): maximum 12.0 per cent, determined
Centrifuge and use the supernatant.
on 1.000 g of the powdered herbal drug (355) (2.9.12) by
Reference solution. Dissolve 1 mg of aescin R and 1 mg of drying in an oven at 105 ºC for 2 h.
glucose R in 1 mL of methanol R. Total ash (2.4.16): maximum 5.0 per cent.
Plate: TLC silica gel plate R (2-10 µm). Ash insoluble in hydrochloric acid (2.8.1): maximum 1.0 per
cent.
Mobile phase: water R, methanol R, methylene chloride R
(10:50:64 V/V/V).
ASSAY
Application: 8 µL as bands of 8 mm. Liquid chromatography (2.2.29).
Development: over a path of 6 cm. Test solution. To 2.000 g of the powdered herbal drug (355)
Drying: in air. (2.9.12) in a round-bottomed flask add 40 mL of a 15 per
cent V/V solution of sulfuric acid R. Heat in a water-bath under
Detection: treat with anisaldehyde solution R, heat at 100 ºC a reflux condenser for 3 h. Allow to cool and filter. Wash the
for 3 min, allow to cool for 1O min and examine in daylight. residue with water R until the filtrate is neutral. Sonicate the
Lavender flower EUROPEAN PHARMACOPOEIA 9.5
residue for 30 min in 80 mL of methanol R and filter. Wash the A. The flower has a short peduncle and consists of a
residue with 20 mL of methanol R, combine the filtrate and the bluish-grey tubular calyx divided distally into 4 very short
washing and dilute to 100.0 mL with methanol R. teeth and a small rounded lobe, a blue bilabial corolla
Reference solution (a). Dissolve 5.0 mg of diosgenin CRS in with the upper lip bifid and the lower lip trilobate and
methanol R and dilute to 25.0 mL with the same solvent. 4 didynamous stamens with ovoid anthers.
B. Microscopic examination (2.8.23). The powder is
Reference solution (b). Dissolve 2 mg of (25R)-spirost-5-en-3-
bluish-grey. Examine under a microscope using chloral
one CRS in reference solution (a) and dilute to 10.0 mL with
the same solution. hydrate solution R. The powder shows the following
diagnostic characters (Figure 1534.-1): covering trichomes
Column: bifurcating at one or more levels [C, L] ; secretory trichomes
- size: l = 0.15 m, 0 = 4.6 mm; with short stalks and 8-celled heads of the Lamiaceae type
(side view [H], surface view [M]); glandular trichomes
- stationary phase: end-capped octadecylsilyl silica gel for with unicellular [O] or multicellular [K] stalks and
chromatography R (5 µm). unicellular heads; glandular trichomes with long uneven
Mobile phase: water for chromatography R, acetonitrile for stalks and unicellular heads, separated from the stalk by an
chromatography R (15:85 V/V). intermediary cell with a smooth cuticle, certain trichomes
show a crown of small spheroid protuberances just below
Flow rate: 1.5 mL/min. the insertion point of the intermediary cell on the stalk [G] ;
Detection: spectrophotometer at 205 nm. fragments of papillose epidermis from the inner surface of
the petals (surface view [J], side view [P]); fragments of
Injection: 5 µL. calyx epidermis with sinuous-walled cells and containing
Run time: 20 min. prismatic crystals of calcium oxalate [Q] ; spherical pollen
Retention time: diosgenin = about 8 min; (25R)-spirost-5-en- grains which have a diameter of about 45 µm and an exine
3-one = about 10 min. with 6 slit-like germinal pores and 6 ribbon-like groins
radiating from the poles [A, D, E, F]; rare fragments of
System suitability: reference solution (b): leaf epidermis with stomata, mostly of the diacytic type
- resolution: mínimum 1.9 between the peaks due to (2.8.3) [B]; fragments of vascular tissue with spiral vessels
diosgenin and (25R)-spirost-5-en-3-one. included in parenchyma with sorne cells containing small
cluster crystals of calcium oxalate [N].
Calculate the percentage content of diosgenin using the
following expression :
Ai x m2 x 4 x p
A2 X m1
area of the peak due to diosgenin in the

chromatogram obtained with the test solution;
area of the peak due to diosgenin in the
chromatogram obtained with reference
w
solution (a);
mass of the herbal drug to be examined used to
prepare the test solution, in grams;
mass of diosgenin CRS used to prepare reference
solution (a), in grams;
p percentage content of diosgenin in diosgenin CRS.
MM
07/2018:1534
J
w
N
LAVENDER FLOWER
Lavandulae flos
t----t
25 µm
DEFINITION
Dried flower of Lavandula angustifolia Mill. (L. officinalis
Chaix). Figure 1534.-1. - Illustration for identification test B of
powdered herbal drug of lavender flower
Content: mínimum 13 mL/kg of essential oil (anhydrous
-
drug).
IDENTIFICATION
First identification: A, B, D.
C. Examine the chromatograms obtained in the test for
lavandin flower.
chromatograms obtained with the reference solution and
the test solution. Furthermore, other faint zones may
Second identification: A, B, C. solution.

EUROPEAN PHARMACOPOEIA 9.5 Lavender oil
Top of the plate Temperature:

Time Temperature
(min) (ºC)
A violet zone o - 15
Column 70
--- --- 15 - 70 70 -7 180
Linalyl acetate: a violet zone A violet zone Injection port 220
A pink zone Detector 220
--- ---
Detection: flame ionisation.
Injection: the same volume of each solution.
1,8-Cineole: a violet or brown Elution arder: limonene, 1,8-cineole, camphor, linalol, linalyl
zone
acetate, a-terpineol.
System suitability: reference solution:
- resolution: minimum 1.5 between the peaks due to
limonene and 1,8-cineole;
Linalol: a violet zone A violet zone (linalol)
- number of theoretical plates: minimum 30 000, calculated
for the peak due to limonene at 110 ºC.
Reference solution Test solution Using the retention times determined from the chromatogram
obtained with the reference solution, locate the 6 components
D. Examine the chromatograms obtained in the test for other of the reference solution in the chromatogram obtained with
species and varieties of lavender. the test solution. Disregard the peaks due to heptane and
Results: the 5 principal peaks in the chromatogram xylene.
obtained with the test solution are similar in retention time Limit:
to the corresponding peaks in the chromatogram obtained - camphor: maximum 1 per cent.
with the reference solution; the 2 main peaks are due to
linalol and linalyl acetate. Water (2.2.13): maximum 100 mL/kg, determined on 20.0 g.
Total ash (2.4.16): maximum 9.0 per cent.
TESTS
Foreign matter (2.8.2): maximum 3 per cent of stems and ASSAY
maximum 2 per cent of other foreign matter. Essential oil (2.8.12). Use 20.0 g of the herbal drug, a 1000 mL
Lavandin flower (Lavandula x intermedia Emeric ex Loisel). round-bottomed flask, 500 mL of water Ras the distillation
liquid and 0.5 mL of xylene R in the graduated tube. Distil at
Thin-layer chromatography (2.2.27).
a rate of 2-3 mL/min for 2 h.
Test solution. To 0.5 g of the powdered herbal drug (355)
(2.9.12) add 5 mL of toluene R, sonicate for 5 min and filter.
Referenr:e .rnlution. Dilute 1O uL of cineole R. 5 uL of linalol R • •: 07/2018:1338
;n'd 5 µL of linalyl acetate R t~ 1 mL with to-lue~e R.
Plate: TLC silica gel F254 plate R (2-10 µm).
Mobile phase: ethyl acetate R, toluene R (5:95 V/V).
Application: 4 µL as bands of 8 mm. LAVENDER 011
Development: over a path of 6 cm.
Drying: in air. Lavandulae aetheroleum
Detection: treat with anisaldehyde solution R and heat at DEFINITION
100 ºC for 5 min; examine immediately in daylight. Essential oil obtained by steam distillation of the flowering
Results: the chromatogram obtained with the test solution tops of Lavandula angustifolia Mill. (Lavandula officinalis
shows no zone between the zones dueto 1,8-cineole and linalol Chaix).
in the chromatogram obtained with the reference solution.
CHARACTERS
Other species and varieties oflavender. Gas chromatography
(2.2.28): use the normalisation procedure. Appearance: colourless or pale yellow, clear liquid.
Test solution. Dilute 0.2 mL of the essential oil-xylene mixture Odour: complex, reminiscent of linalyl aceta te.
obtained in the assay to 5 mL with heptane R, add 1 g of IDENTIFICATION
anhydrous sodium sulfate R, shake and use the supernatant.
First identification: B.
Reference solution. Dissolve O.OS g of camphor R and 0.2 g of
Second identification: A.
a-terpineol R in heptane R, add 0.1 g of limonene R, 0.2 g of
cineole R, 0.4 g of linalol R and 0.6 g of linalyl acetate R, then A. Thin-layer chromatography (2.2.27).
dilute to 100 mL with heptane R. Test solution. Dilute 20 µL of the essential oil to be
Column: examined to 1 mL with toluene R.
Reference solution. Dilute 10 µL of cineole R, 5 µL of
- material: fused silica;
linalol R and 5 µL of linalyl acetate R to 1 mL with toluene R.
- size: l = 60 m, 0 = 0.25 mm; Plate: TLC silica gel F254 plate R (5-40 µm) [or TLC silica
- stationary phase: macrogol 20 000 R (film thickness gel F254 plate R (2-10 µm)].
0.25 µm). Mobile phase: ethyl acetate R, toluene R (5:95 V/V).
Carrier gas: helium for chromatography R. Application: 6 µL [or 2 µL] as bands of 10 mm [or 8 mm].
Flow rate: 1.5 mL/min. Development: overa path of 10 cm [or 6 cm].
Split ratio: 1:100. Drying: in air.
Lavender oil EUROPEAN PHARMACOPOEIA 9.S
Detection: treat with anisaldehyde solution R and heat at Injection: 1 µL.

100 ºC for S min; examine immediately in daylight. Elution arder: limonene, 1,8-cineole, 3-octanone, camphor,
Results: see below the sequence of zones present in the linalol, linalyl acetate, terpinen-4-ol, lavandulyl acetate,
chromatograms obtained with the reference solution and lavandulol, a-terpineol.
the test solution. Furthermore, other faint zones may Identification of peaks: use the chromatogram supplied with
be present in the chromatogram obtained with the test lavender oil for peak identification HRS and the chromatogram
solution. obtained with reference solution (a) to identify the peaks
Top of the plate
dueto limonene, 1,8-cineole, 3-octanone, camphor, linalol,
linalyl acetate, terpinen-4-ol, lavandulyl acetate, lavandulol
A violet zone and a-terpineol.
--- --- System suitability: reference solution (a):
-
Linalyl acetate: a violet zone An intense violet zone (linalyl - resolution: minimum 1.4 between the peaks due to
acetate) terpinen-4-ol and lavandulyl acetate.
A pink zone
Determine the percentage content of each of the following
components. The percentages are within the following ranges:
--- --- - limonene: maximum 1.0 per cent;
1,8-Cineole: a violet or brown Possibly a weak violet-brown - 1,8-cineole: maximum 2.S per cent;
zone zone (1,8-cineole)
- 3-octanone: 0.1 per cent to S.O per cent;
- camphor: maximum 1.2 per cent;
Linalol: a violet zone An intense violet zone (linalol)
- linalol: 20.0 per cent to 4S.O per cent;
A greyish or brownish zone
- linalyl acetate: 2S.O per cent to 47.0 per cent;
- terpinen-4-ol: 0.1 per cent to 8.0 per cent;
Reference solution Test solution - lavandulyl acetate: minimum 0.2 per cent;
- lavandulol: minimum 0.1 per cent;
B. The essential oil to be examined complies with the limits
of the test for chromatographic profile. - a-terpineol: maximum 2.0 per cent;
- reporting threshold: the area of the principal peak in
TESTS the chromatogram obtained with reference solution (b)
Relative density (2.2.5): 0.878 to 0.892. (O.OS per cent).
Refractive index (2.2.6): l.4SS to 1.466. Chiral purity. Gas chromatography (2.2.28).
Test solution. Dilute 0.02 g of the essential oil to be examined
Optical rotation (2.2.7): - 12.Sº to - 6.0º.
to 10 mL with pentane R.
Acid value (2.5.1): maximum 1.0, determined on S.00 g of Reference solution. Dissolve S mg of borneo[ R in pentane R,
the essential oil to be examined dissolved in SO mL of the add 10 µL of linalol R (mixture of (R)-linalol and (S)-linalol)
prescribed mixture of solvents. and 10 µL of linalyl acetate R (mixture of (R)-linalyl acetate
Chromatographic profile. Gas chromatography (2.2.28): use and (S)-linalyl acetate) and dilute to 10 mL with pentane R.
the normalisation procedure. Column:
Test solution. Dilute 200 µL of the essential oil to be examined - material: fused silica;
to 10.0 mL with heptane R.
- size: l = 2S m, 0 = 0.2S mm;
Reference solution (a). Dilute 200 µL of lavender oil for peak
- stationary phase: modified [3-cyclodextrin for chiral
identification HRS to 10.0 mL with heptane R.
chromatography R (film thickness 0.2S µm).
Reference solution (b). Dilute S µL of limonene R to SO.O mL
Carrier gas: helium f or chromatography R.
with heptane R. Dilute O.S mL of this solution to S.O mL with
heptane R. Flow rate: 1.3 mL/min.
Column: Split ratio: 1 :30.
- material: fused silica; Temperature:
- size: l = 60 m, 0 = 0.2S mm; Time Temperature
(min) (ºC)
stationary phase: macrogol 20 000 R (film thickness
Column o - 65 50 7 180
0.2S µm).
Injection port 230
Carrier gas: helium for chromatography R.
Flow rate: l.S mL/min. Detector 230
Split ratio: 1 :SO. Detection: flame ionisation.

Temperature: Injection: 1 µL.
Time Temperature Elution arder: (R)-linalol, (S)-linalol, borneol, (R)-linalyl
(min) (ºC) acetate, (S)-linalyl acetate; depending on the operating
Column o - 15 70 conditions and the state of the column, borneol may elute
before or after (S)-linalol.
15 - 70 70 7 180
Injection port 220
- resolution: minimum S.S between the peaks due to
Detector 220 (R)-linalol and (S)-linalol; minimum 2.9 between the peaks
dueto (S)-linalol and borneol; minimum 2.0 between the
Detection: flame ionisation. peaks dueto (R)-linalyl acetate and (S)-linalyl acetate.
S616 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.5 Spike lavender oil
Calculate the percentage content of the specified Top of the plate

(S)-enantiomers using the following expression: A violet zone
As --- ---
A
s+ A R X 100
A faint violet or brown zone may
Linalyl aceta te: a violet zone
be present (linalyl acetate)
A5 area of the peak dueto the corresponding A pink zone
(S)-enantiomer;
--- ---
AR area of the peak due to the corresponding
(R)-enantiomer.
Limits: 1,8-Cineole: a violet or brown A violet or brown zone
zone (1,8-cineole)
- (S)-linalol: maximum 12 per cent;
- (S)-linalyl acetate: maximum 1 per cent. Linalol: a violet zone An intense violet zone (linalol)
A greyish or brownish zone

STORAGE
At a temperature not exceeding 25 ºC.
Reference solution Test solution
B. The essential oil to be examined complies with the limits

of the test for chromatographic profile.
07/2018:2419 TESTS
Relative density (2.2.5): 0.894 to 0.907.
Refractive index (2.2.6): 1.461 to 1.468.
Optical rotation (2.2. 7): - 7º to + 2º.
SPIKE LAVENDER OIL Acid value (2.5.1): maximum 1.5, determined on 5.00 g ofthe
essential oil to be examined.
Solubility in alcohol (2.8.10): 1.0 mL of the essential oil to be
Spicae aetheroleum examined is soluble, sometimes with opalescence, in 3.0 mL of
ethanol (70 per cent V/V) R.
DEFINITION Chromatographic profile. Gas chromatography (2.2.28): use
Essential oil obtained by steam distillation of the flowering the normalisation procedure.
tops of Lavandula latifolia Medik. Test solution. Dilute 200 µL of the essential oil to be examined
CHARACTERS Reference solution (a). Dilute 200 µL of spike lavender oíl CRS
Appearance: clear, mobile, light yellow or greenish-yellow Reference solution (b). Dilute 5 µL of limonene R to SO.O mL
liquid. with heptane R. Dilute 0.5 mL of this solution to 5.0 mL with
Odour: reminiscent of cineole and camphor. heptane R.
Column:
IDENTIFICATION - material: fused silica;
First identification: B. - size: l = 60 m, 0 = 0.25 mm;
- stationary phase: macrogol 20 000 R (film thickness
Second identification: A. 0.25 µm).
A. Thin-layer chromatography (2.2.27). Carrier gas: helium far chromatography R.
Test solution. Dilute 20 µL of the essential oil to be Flow rate: 1.5 mL/min.
examined to 1 mL with toluene R. Split ratio: 1:50.
Reference solution. Dilute 10 µL of cineole R, 5 µL of Temperature:
linalol R and 5 µL of linalyl acetate R to 1 mL with toluene R. Time Temperature
Plate: TIC sílica gel F254 plate R (5-40 µm) [or TLC sílica (min) (ºC)
gel F254 plate R (2-10 µm)]. Column o - 15 70
15 - 70 70 ~ 180
Mobíle phase: ethyl acetate R, toluene R (5:95 V/V). Injection port 220
Application: 6 µL [or 2 µL] as bands of 10 mm [or 8 mm]. Detector 220
Development: overa path of 10 cm [or 6 cm].
Drying: in air.
Injection: 1 µL.
Detection: treat with anisaldehyde solution R and heat at Elution arder: limonene, 1,8-cineole, camphor, linalol, linalyl
100 ºC for 5 min; examine immediately in daylight. acetate, a-terpineol, trans-a-bisabolene.
Results: see below the sequence of zones present in the Identification of peaks: use the chromatogram supplied with
chromatograms obtained with the reference solution and spike lavender oíl CRS and the chromatogram obtained with
the test solution. Furthermore, other faint zones may reference solution (a) to identify the peaks dueto limonene,
be present in the chromatogram obtained with the test 1,8-cineole, camphor, linalol, linalyl acetate, a-terpineol and
solution. trans-a-bisabolene.
Spike lavender oil EUROPEAN PHARMACOPOEIA 9.S
System suitability: reference solution (a): - linalol: 34.0 per cent to SO.O per cent;
- the chromatogram obtained is similar to the chromatogram linalyl acetate: maximum 1.6 per cent;
supplied with spike lavender oíl CRS; - a-terpineol: 0.2 per cent to 2.0 per cent;
- resolution: mínimum 1.S between the peaks due to - trans-a-bisabolene: 0.4 per cent to 2.5 per cent;
limonene and 1,8-cineole.
- reporting threshold: the area of the principal peak in
Determine the percentage content of each of the following
the chromatogram obtained with reference solution (b)
components. The percentages are within the following ranges:
(O.OS per cent).
- limonene: O.S per cent to 3.0 per cent;
- 1,8-cineole: 16.0 per cent to 39.0 per cent; STORAGE
- camphor: 8.0 per cent to 16.0 per cent; At a temperature not exceeding 2S ºC.

Homoeopathic preparations
Acidum succinicum for homoeopathic preparations ........ 5621 Aurum chloratum natronatum for homoeopathic
Agaricus phalloides for homoeopathic preparations ......... 5621 preparations .......................................................................... 5623
Arsenicum album for homoeopathic preparations ............ 5623 Calcium fluoratum for homoeopathic preparations.......... 5624

EUROPEAN PHARMACOPOEIA 9.5 Agaricus phalloides for homoeopathic preparations
07/2018:2824 07/2018:2290
~
E
ACIDUM SUCCINICUM FOR AGARICUS PHALLOIDES FOR
HOMOEOPATHIC PREPARATIONS(I) HOMOEOPATHIC PREPARATIONS<2>
Acidum succinicum ad praeparationes Amanita phalloides ad praeparationes

homoeopathicas homoeopathicas
DEFINITION
Whole, fresh mushroom (fruiting body) Amanita phalloides
C4H604 Mr 118.1 (Vaill. ex Fr.) Link.
[110-15-6]
IDENTIFICATION
DEFINITION
A. In young specimens, the cap, 50-150 mm in diameter, is
Butanedioic acid (succinic acid). hemispherical to campanulate with margins rolled inwards
Content: 99.0 per cent to 101.0 per cent. and is still covered by the white universal veil; in mature
specimens, the cap is expanded, umbrella-like, convex to
CHARACTERS nearly plane and occasionally depressed; its colour is pale
green along the margin and elsewhere greyish-green to
Appearance: white or almost white, crystalline powder or yellowish-green, typically with fine grey streaks growing
colourless crystals. thicker towards the centre; the cuticle is peelable up to
Solubility: soluble in water and in ethanol (96 per cent), the middle of the cap as a fine membrane; the underlying
sparingly soluble in acetone. flesh appears greenish-yellow, becoming more intense
towards the centre of the cap, while in the other parts of
the mushroom the flesh is uniformly whitish; lamellae
IDENTIFICATION are free, closely packed, white with a slight yellowish
First identification: C. tinge; lamellulae are truncate; the stipe, about 1O cm high
and 2 cm in diameter, is thin and solid, with a whitish,
Second identification: A, B, D. membranous, striate annulus and an enlarged and bulbous
A. Solution S (see Tests) is strongly acid (2.2.4). base that almost always shows a tough, white, membranous
sac-like volva torn into irregular lobes at the top. In old
Meltirn1 noint (2.2.14):
R - -------o e - ' , 184 ºC to 187 ºC.
--
specimens, the stipe is hollow, whitish and often covered
C. Infrared absorption spectrophotometry (2.2.24). with small greenish scales around the annulus. The spores
are white.
Comparíson: succinic acíd CRS.
B. Examined under a lens (xlO), the upper surface is shiny,
D. Neutralise 3 mL of solution S with 1 M sodium hydroxide, dry, appearing somewhat uneven, with no remains of the
then add 1 mL of silver nítrate solution R2. A white
veil.
precipitate is formed.
C. Examine under a microscope using a solution containing
TESTS 1.5 g of iodine R, 5 g of potassium iodide R and 100 g
of chloral hydrate R in 100 mL of water R. The spores
Solution S. Dissolve 5.0 g in distilled water R and dilute to are blackish-blue (starch reaction), short elliptical to
100 mL with the same solvent. subspherical, 8-11 µm long and 7-9 µm in diameter.
Chlorides (2.4.4): maximum 100 ppm.
Dilute 10 mL of solution Sto 15 mL with water R. TESTS
Sulfates (2.4.13): maximum 200 ppm, determined on Foreign matter (2.8.2): maximum 5 per cent.
solution S. Loss on drying (2.2.32): minimum 85.0 per cent, determined
Oxalates. Neutralise 5 mL of solution S with dilute on 5.0 g of the finely cut drug by drying in an oven at 105 ºC
ammonia Rl, using 0.1 mL of phenolphthaleín solutíon R as for 2 h.
indicator. Add 0.1 mL of acetic acid R and 5 mL of calcíum Other Amanita species. Veil remnants on caps are typical for
sulfate solution R. The solution remains clear for at least most Amanita species, but not for A. phalloides. Therefore
20 min. all mushrooms with veil remnants on the cap have to be
discarded. The presence of veil remnants (patches) on the
ASSAY cap indicates adulteration with A. citrina (Schaeff.) Pers.
(whitish-yellow cap with whitish to brownish patches) or
Dissolve 0.500 gin 50 mL of water R. Titrate with 1 M sodium
with A. muscaria (L.: Fr.) Lamarck, A. caesarea (Scop.: Fr.)
hydroxide, determining the end-point potentiometrically
Pers., or A. rubescens Pers. (orange to bright red cap with
(2.2.20) or using 0.2 mL of phenolphthalein solution R as
white patches); a brownish cap indicates adulteration with A.
indicator.
pantherína (DC.) Krombh.; a greenish-white cap anda white
1 mL of 1 M sodium hydroxide is equivalent to 59.05 mg of stipe with a labile annulus indicates adulteration with A. verna
C4H604. (Bull.: Fr.) Lamarck.
(1) FRENCH TITLE: Succinicum acidum pour préparations homéopathiques

(2) FRENCH TITLE: Agaricus bulbosus pour préparations homéopathiques
Agaricus phalloides for homoeopathic preparations EUROPEAN PHARMACOPOEIA 9.5
Mother tincture B. Thin-layer chromatography (2.2.27).

The mother tincture complies with the requirements of Test solution. The mother tincture to be examined.
the general monograph Mother tinctures for homoeopathic Reference solution. Dissolve 2 mg of gramine R and 2 mg
preparations (2029). of rutoside trihydrate R in methanol R and dilute to 1O mL
with the same solvent.
DEFINITION Plate: TLC silica gel plate R (5-40 µm) [or TLC silica gel
Content: 0.001 per cent m/m to 0.010 per cent m!m for the F254 plate R (2-10 µm)].
sum of a-amanitine and ~-amanitine (C39 H 54 N 100 14S; Mr 919). Mobile phase: glacial acetic acid R, water R, methanol R,
PRODUCTION methylene chloride R (4:6:30:60 V/V/V/V).
The mother tincture is prepared according to the following Application: 40 µL [or 10 µL] as bands.
methods as prescribed in the monograph Methods of Development: overa path of 10 cm [or 6 cm].
preparation of homoeopathic stocks and potentisation (2371): Drying: in a current of warm air.
- method 1.1.5; Detection: treat with a 5 per cent V/V solution of cinnamic
- method 1.1.1 O, using 5 parts of the cut drug for 100 parts aldehyde R in methanol R and expose to hydrochloric acid R
of ethanol (45 per cent V!V) and maceration for 3 weeks. vapour for 30 min; examine in daylight.
CHARACTERS chromatograms obtained with the reference solution and
Appearance: brownish-yellow, yellowish or green liquid. the test solution. Furthermore, other faint zones may
IDENTIFICATION solution.
Carry out test A when method 1.1.5 is used and carry out
Top of the plate
test B when method 1.1.10 is used.
A. Thin-layer chromatography (2.2.27). --- ---
Test solution. Evaporate to dryness 20 mL of the mother Gramine: a yellow zone

tincture under vacuum at about 40 ºC. Dissolve the residue
Rutoside: a yellow zone
in 1 mL of water R and add 1 mL of methanol R. Filter
immediately. A pinkzone
Reference solution. Dissolve 10 mg of rutoside trihydrate R A pinkzone
and 1O mg of sennoside B R in methanol R and dilute to
40 mL with the same solvent. --- ---
Plate: TLC silica gel F254 plate R (5-40 µm) [or TLC silica An orange-yellow zone
gel F254 plate R (2-10 µm)].
Reference solution Test solution
Mobile phase: glacial acetic acid R, water R, butano[ R
(17:17:66 V/V/V).
TESTS
Application: 40 µL [or 10 µLJ as bands.
Relative density (2.2.5): 0.895 to 0.915, where method 1.1.5
Development: overa path of 10 cm [or 6 cm].
is used.
Drying: in a current of warm air.
Ethanol (2.9.10): 40 per cent V/V to 50 per cent V/V, where
Detection A: examine in ultraviolet light at 254 nm. method 1.1.1 O is used.
Results A: locate a quenching zone (sennoside B) in the Dry residue (2.8.16): minimum 0.8 per cent.
lower third anda quenching zone (rutoside) in the middle
third of the chromatogram obtained with the reference Mother tincture of Agaricus muscarius. Thin-layer
solution. chromatography (2.2.27).
Detection B: treat immediately with a 1 per cent V/V Test solution. The mother tincture to be examined.
solution of cinnamic aldehyde R in methanol R and allow to Reference solution. Dissolve 10 mg of leucine R and 10 mg
dry; treat with hydrochloric acid R; examine in daylight. of threonine R in 5 mL of water R and dilute to 20 mL with
Results B: see below the sequence of zones present in the ethanol (96 per cent) R.
chromatograms obtained with the reference solution and Plate: TLC silica gel plate R (5-40 µm) [or TLC silica gel plate R
the test solution. Furthermore, other faint zones may (2-10 µm)J.
be present in the chromatogram obtained with the test Mobile phase: glacial acetic acid R, water R, acetone R,
solution. butano[ R (10:20:35:35 V/V/V/V).
Top of the plate Application: 20 µL [or 10 µLJ as bands.
--- --- --- Development: overa path of 10 cm [or 6 cm].
Drying: in a current of warm air.
Rutoside: a Rutoside: a yellow
quenching zone zone Detection: treat with a 1 g/L solution of ninhydrin R in
A violet zone butano[ R and heat at 105 ºC for 5-10 min; examine in daylight.
Results: the presence of noticeable zones in the chromatogram
obtained with the test solution, in the same position as the
--- --- --- zones due to leucine and threonine in the chromatogram
Sennoside B: a A violet zone obtained with the reference solution, indicates adulteration
quenching zone with mother tincture of Agaricus muscarius.
A violet zone
ASSAY
2 faint greyish-violet Liquid chromatography (2.2.29).
zones may be present
Test solution. Evaporate 2.000 g of the mother tincture to
be examined to dryness. Dissolve the residue in 2.0 mL of
Reference solution Reference solution Test solution water for chromatography R. Filter through a membrane filter
(detection A) (detection B) (detection B) (nominal pore size 0.45 µm).

EUROPEAN PHARMACOPOEIA 9.5 Aurum chloratum natronatum for homoeopathic preparations
Reference solution. Dissolve 10.0 mg of tryptophan CRS in DEFINITION

mobile phase A and dilute to 20.0 mL with mobile phase A. Content: 99.5 per cent to 100.5 per cent of AszÜ 3•
Column:
- size: l = 0.10 m, 0 = 4.6 mm; CHARACTERS
- stationary phase: end-capped so lid core octadecylsilyl silica Appearance: white or almost white powder.
gel for chromatography R (2.6 µm); Solubility: practically insoluble to sparingly soluble in water.
- temperature: 35 ºC. It dissolves in solutions of alkali hydroxides and carbonates.
Mobile phase: IDENTIFICATION
- mobile phase A: dissolve 1.54 g of ammonium acetate R in A. Dissolve 20 mg in 1 mL of dilute hydrochloric acid R, add
900 mL of water for chromatography R, adjust to pH 5.0 4 mL of water R and 0.1 mL of sodium sulfide solution R.
with glacial acetic acid R and dilute to 1 L with water for The resulting yellow precipitate is soluble in dilute
chromatography R; ammonia Rl.
- mobile phase B: acetonitrile for chromatography R; B. Dissolve 20 mg in 1 mL of hydrochloric acid Rl, add 5 mL
Time Mobile phase A Mobile phase B of hypophosphorous reagent R and heat for 15 min on a
(min) (per cent VIV) (per cent V/V) water-bath. A black precipitate develops.
0-2 95 5
TESTS
2 - 17 95 7 80 5 7 20 Appearance of solution. The solution is clear (2.2.1) and
17 - 22 80 7 50 20 -7 50 colourless (2.2.2, Method 11).
Prepare a 100 g/L solution in dilute ammonia Rl, heating if
Flow rate: 1.0 mL/min. necessary.
Detection: spectrophotometer at 303 nm. Sulfides: maximum 20 ppm.
1njection: 10 µL. Dissolve 1.0 gin 10.0 mL of dilute sodium hydroxide solution R.
Relative retention with reference to tryptophan (retention Add 0.05 mL of lead acetate solution R. Any colour in the test
time = about 3 min): ~-amanitine = about 2.9; solution is not more intense than that in a standard prepared
a-amanitine = about 3.2. at the same time and in the same manner using a mixture of
System suitability: test solution: 10.0 mL of a 0.015 g/L solution of sodium sulfide R in dilute
- resolution: minimum 2.0 between the peaks due to sodium hydroxide solution R and 0.05 mL of lead acetate
~-amanitine and a-amanitine.
solution R.
Calculate the percentage content m!m of a-amanitine and ASSAY
~-amanitine using the following expression:
Dissolve 40.0 mg in a mixture of 1O mL of dilute sodium
(A2 + A3) x m1 xpx K x O. l hydroxide solution R and 10 mL of water R. Add 10 mL
A1 X m2 of dilute hydrochloric acid R and 3 g of sodium hydrogen
carbonate R and mix. Add 1 mL of starch solution R and titrate
area of the peak due to tryptophan in the with O. 05 M iodine.
chromatogram obtained with the reference l mL of 0.05 M iodine is equivalent to 4.946 mg of As 203"
solution;
area of the peak dueto a-amanitine in the 07/2018:2141
area of the peak due to ~-amanitine in the
mass of tryptophan CRS used to prepare the AURUM CHLORATUM NATRONATUM
reference solution, in grams;
mass of the mother tincture to be examined
FOR HOMOEOPATHIC
used to prepare the test solution, in grams; PREPARATIONS<4J
p percentage content of tryptophan in
tryptophan CRS; Natrii tetrachloroauras dihydricus ad
K correction factor between a-amanitine, praeparationes homoeopathicas
~-amanitine and tryptophan (0.1).
Na[AuCl1 ],2HzÜ Mr 397.8
[13874-02-7]
07/2018:1599
DEFINITION
Sodium tetrachloroaurate( 1-) dihydrate.
Content: 97.0 per cent to 101.0 per cent of Na[AuC14 ],2HzÜ.
ARSENICUM ALBUM FOR CHARACTERS
HOMOEOPATHIC PREPARATIONS(3) Appearance: orange-yellow, hygroscopic powder or crystals.
Solubility: very soluble or freely soluble in water and in
ethanol (96 per cent).
Arsenii trioxidum ad praeparationes
homoeopathicas IDENTIFICATION
A. Dissolve 20 mg in 2.0 mL of 0.1 M nitric acid. Add 0.1 g
As2 0 3 Mr 197.8 of oxalic acid R and boil in a water-bath for 1 h. A deposit
[1327-53-3] of metallic gold is formed.
(3) FRENCH TITLE: Arsenicum album pour préparations homéopathiques

(4) FRENCH TITLE: Aurum muriaticum natronatum pour préparations homéopathiques
General Notíces (1) apply to ali monographs and other texts 5623
Calcium fluoratum for homoeopathic preparations EUROPEAN PHARMACOPOEIA 9.5
B. Solution S (see Tests) gives reaction (a) of chlorides (2.3.1). 07/2018:2996

C. Solution S gives reaction (b) of sodium (2.3.1 ).
TESTS
Solution S. Ignite 0.20 g in a porcelain crucible at
600 ºC ± 50 ºC for 30 min. Allow to cool and extract with
3 mL of water R, heating if necessary. Use the supernatant.
CALCIUM FLUORATUM FOR
Free hydrochloric acid. When a glass rod impregnated with
HOMOEOPATHIC PREPARATIONS<sJ
concentrated ammonia R is held close to the substance to be
examined, no white fumes are produced. Calcii fluoridum ad praeparationes
Nitrates: maximum 200 ppm. homoeopathicas
Dissolve 0.20 g in 4.0 mL of nitrate-free water R. Add 0.6 g of
zinc R and 1O mL of dilute sulfuric acid R. Heat the solution
on a water-bath for 30 min, allow to cool and filter. Rinse CaF 2 Mr 78.1
the filter with nitrate-free water R and dilute the filtrate to [7789-75-5]
20.0 mL with the same solvent. To 5.0 mL of this solution add
0.4 mL of a 100 g/L solution of potassium chloride R, 0.1 mL DEFINITION
of diphenylamine solution R and, dropwise with shaking,
5.0 mL of nitrogen-free sulfuric acid R. Heat in a water-bath Content: 98.0 per cent to 102.0 per cent of CaF 2 •
at 50 ºC for 15 min, centrifuge if necessary and use the clear
supernatant. Any blue colour in the solution is not more CHARACTERS
intense than that in a reference solution prepared at the same
time and in the same manner using a mixture of 1.0 mL Appearance: fine, white or almost white powder.
of nitrate standard solution (10 ppm NO) R and 3.0 mL of Solubility: practically insoluble in water, slightly soluble in
nitrate-free water R. mineral acids.
ASSAY
IDENTIFICATION
Dissolve 40.0 mg in 10 mL of potassium iodide solution R.
Allow to stand for 5 min. Titrate with 0.01 M sodium A. To 0.80 g add 20 mL of hydrochloric acid R and heat
thiosulfate until decolourised. Shortly before reaching the to boiling under a reflux condenser until complete
endpoint, add 0.5 mL of starch solution R. dissolution (about 30 min). After cooling, add 0.1 mL
1 mL of 0.01 M sodium íhiosulfaíe is equivaient to i.989 mg oí of phenolphthalein solution R, and then concentrated
Na[AuC14 ],2Hp. ammonia R until a pink colour is obtained. Add glacial
acetic acid R until the solution is decolourised, then add
STORAGE 1 mL in excess. Filter and dilute to 40 mL with water R.
Dilute 1 mL of the solution obtained to 5 mL with distilled
In an airtight container, protected from light.
water R and add 2 mL of ammonium oxalate solution R.
A white precipitate is formed which dissolves in 2 mL of
dilute hydrochloric acid R.
B. In a lead or platinum crucible, mix 1O mg with 20 mg of
anhydrous colloidal silica R and a few drops of sulfuric
acid R, with the aid of a copper wire, in order to give a thin
slurry. Cover the crucible with a thin, transparent plate of
plastic under which a drop of water R is suspended, and
warm gently. A white ring is rapidly formed around the
drop of water.
TESTS
Free acid. Shake 5.0 g with 2 g of calcium chloride R and
100 mL of water R for 5 min. Heat to 70 ºC and filter. To 40 mL
of the filtrate, maintained at 70 ºC, add 0.1 mL of methyl red
solution R. Not more than 1.0 mL of 0.1 M sodium hydroxide
is required to change the colour of the indicator to yellow.
ASSAY
Introduce 0.150 g into a 500 mL conical flask and add 8 mL
of hydrochloric acid R. Boil for 3-4 min on a preheated hot
plate and allow to cool. Add 300 mL of water R, followed by
strong sodium hydroxide solution R until the first appearance
of persistent opalescence (about pH 14). Add 0.13 g of
calconecarboxylic acid tríturate R and titrate with 0.1 M sodium
edetate until the colour changes from red-violet to pure blue.
The opalescence caused by the strong sodium hydroxide
solution disappears during the course of the titration. If still
visible at the end of the titration, it can be dissolved by adding
a few drops of hydrochloric acid R.
1 mL of 0.1 M sodium edetate is equivalent to 7.81 mg of CaF 2•
(5) FRENCH TJTLE: Calcarea fluorica pour préparations homéopathiques

A
Acitretin ................................................................................... 5627

EUROPEAN PHARMACOPOEIA 9.5 Acitretin
07/2018:1385 - temperature: 25 ºC.

Mobile phase: 0.3 per cent V/V solution of glacial acetic acid R
in a mixture of 8 volumes of water for chromatography R and
92 volumes of anhydrous ethanol R.
Flow rate: 0.6 mL/min.
ACITRETIN Detection: spectrophotometer at 360 nm.
Autosampler: set at 4 ºC.
Acitretinum Injection: 10 µL of test solution (a) and reference solutions (b ),
«
(c) and (d).
~~C02H
3
CH Run time: 2.5 times the retention time of acitretin.
1
ldentification of impurities: use the chromatogram supplied
H3 CO with acitretin for impurity A identification CRS and the
CH 3
chromatogram obtained with reference solution (d) to identify
the peak due to impurity A.
C21H2603 Relative retention with reference to acitretin (retention
[55079-83-9] time = about 6 min): impurity A = about 0.8;
DEFINITION tretinoin = about 0.85.
System suitability: reference solution (b):
(2E,4E,6E,8E)-9-(4-Methoxy-2,3,6-trimethylphenyl)-3,7-
dimethylnona-2,4,6,8-tetraenoic acid - resolution: mínimum 2.0 between the peaks dueto tretinoin
Content: 98.0 per cent to 102.0 per cent (dried substance).
and acitretin.
Calculation of percentage contents:
CHARACTERS - for each impurity, use the concentration of acitretin in
Appearance: yellow or greenish-yellow, crystalline powder. reference solution (c).
Solubility: practically insoluble in water, sparingly soluble in Limits:
tetrahydrofuran, slightly soluble in acetone and in ethanol - impurity A: maximum 0.2 per cent;
(96 per cent), very slightly soluble in cyclohexane.
- unspecified impurities: for each impuríty, maximum
It is sensitive to air, heat and light, especially in solution. 0.10 per cent;
It shows polymorphism (5.9). - total: maximum 0.5 per cent;
Carry out all operations as rapidly as possible and avoid
exposure to actinic light; use freshly prepared solutions.
IDENTIFICATION
Infrared absorption spectrophotometry (2.2.24).
- reporting threshold: 0.05 per cent.
Loss on drying (2.2.32): maximum 0.5 per cent, determined

on 1.000 g by drying in vacuo at 100 ºC for 4 h.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
-
Comparison: acitretin CRS.
1.0 g.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference ASSAY
substance separately in 2-propanol R by heating under reflux; Liquid chromatography (2.2.29) as described in the test for
filter, evaporate to dryness and record new spectra using the related substances with the following modification.
residues.
lnjection: test solution (b) and reference solution (a).
TESTS Calculate the percentage content of C 21 H 26 Ü 3 taking into
Related substances. Liquid chromatography (2.2.29). Carry account the assigned content of acitretin CRS.
out the test protected from light and prepare the solutions
STORAGE
immediately befare use.
In an airtight container, protected from light, at a temperature
Test solution (a). Dissolve 25.0 mg of the substance to be
of 2 ºC to 8 ºC.
examined in 5 mL of tetrahydrofuran R and dilute to 100.0 mL
with anhydrous ethanol R. It is recommended that the contents of an opened container
be used as soon as possible and any unused part be protected
Test solution (b). Dilute 10.0 mL of test solution (a) to 25.0 mL
by an atmosphere of inert gas.
with anhydrous ethanol R.
Reference solution (a). Dissolve 25.0 mg of acitretin CRS IMPURITIES
in 5 mL of tetrahydrofuran R and dilute to 100.0 mL with Specified impurities: A.
anhydrous ethanol R. Dilute 1O.O mL of the solution to 25.0 mL Other detectable impurities (the following substances would,
with anhydrous ethanol R. if present at a sufficient level, be detected by one or other of
Reference solution (b ). Dissolve 1 mg of tretinoin CRS in the tests in the monograph. They are limited by the general
anhydrous ethanol R and dilute to 20.0 mL with the same acceptance criterion for other/unspecified impurities and/or
solvent. Mix 5.0 mL of the solution with 2.5 mL of reference by the general monograph Substances far pharmaceutical
solution (a) and dilute to 100.0 mL with anhydrous ethanol R. use (2034). It is therefore not necessary to identify these
Reference solution ( c). Dilute 1.0 mL of the test solution (a) impurities for demonstration of compliance. See also 5.1 O.
to 100.0 mL with anhydrous ethanol R. Dilute 1.0 mL of this Control of impurities in substances far pharmaceutical use): B.
solution to 10.0 mL with anhydrous ethanol R.
Reference solution (d). Dissolve 2.5 mg of acitretin for
impurity A identification CRS in 0.5 mL of tetrahydrofuran R
and dilute to 10.0 mL with anhydrous ethanol R.
Column:
- size:l=0.25m,0=4mm;
- stationary phase: octadecylsilyl silica gel for chromatography A. (2Z,4E,6E,8E)-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-
for separation ofpolycyclic aromatic hydrocarbons R (5 µm); dimethylnona-2,4,6,8-tetraenoic acid,
Acitretin EUROPEAN PHARMACOPOEIA 9.5
~0/'-..CH3
H3coYcH3
CH 3
B. ethyl (2E,4E,6E,8E)-9-(4-methoxy-2,3,6-trimethylphenyl)-
3, 7-dimethylnona-2,4,6,8-tetraenoate.

B
Biotin ........................................................................................ 5631

EUROPEAN PHARMACOPOEIA 9.5 Biotin
07/2018:1073 Solvent mixture: water R, acetonitrile R (50:50 V/V).

Test solution. Dissolve 50 mg of the substance to be examined
in the solvent mixture using sonication and dilute to SO.O mL
with the solvent mixture.
Reference solution (a). Dilute 1.0 mL of the test solution to
BIOTIN 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
solution to 1O.O mL with the solvent mixture.
Biotinum Reference solution (b ). Dissolve 5 mg of biotin for system
suitability CRS (containing impurities A, C and E) in the
solvent mixture and dilute to 5.0 mL with the solvent mixture.
Column:
- size: l = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped octadecylsilyl silica gel far
chromatography R (5 µm);
C10H16N203S
[58-85-5] - temperature: 30 ºC.
Mobile phase:
DEFINITION
- mobile phase A: methanesulfonic acid R, acetonitrile Rl,
5-[(3aS,4S,6aR)-2-0xohexahydro-lH-thieno[3,4-d]imidazol- water for chromatography R ( 1:25:1000 V/V/V);
4-yl]pentanoic acid.
- mobile phase B: methanesulfonic acid R, water for
Content: 98.5 per cent to 101.0 per cent (dried substance). chromatography R, acetonitrile Rl (1:25:1000 V/V/V);
CHARACTERS Time Mobile phase A Mobile phase B
Appearance: white or almost white, crystalline powder or (min) (per cent V/V) (per cent V/V)
colourless crystals. o- 5 95 5
Solubility: very slightly soluble in water and in ethanol (96 per 5 - 20 95 -7 o 5 -7 100
cent), practically insoluble in acetone. It dissolves in dilute
solutions of alkali hydroxides. 20 - 28 o 100
IDENTIFICATION Flow rate: 1.0 mL/min.

First identification: A. Detection: spectrophotometer at 200 nm from O to 5 min and
Second identification: B. at 210 nm from 5 to 28 min.
A. Infrared absorption spectrophotometry (2.2.24). Injection: 10 µL.
Comparison: biotin CRS. Identification of impurities: use the chromatogram supplied
with biotin for system suitability CRS and the chromatogram
B. Thin-layer chromatography (2.2.27). obtained with reference solution (b) to identify the peaks due
Prepare the solutions immediately before use and keep to impurities A, C and E.
protected from bright light. Relative retention \Vith reference to biotin (retention
Test solution. Dissolve 5 mg of the substance to be time = about 12 min): impurity e = about 0.2;
examined in glacial acetic acid R and dilute to 10 mL with impurity A = about 1.1; impurity E = about 1.3.
the same solvent.
Reference solution. Dissolve 5 mg of biotin CRS in glacial - resolution: minimum 1.5 between the peaks due to biotin
acetic acid R and dilute to 10 mL with the same solvent. and impurity A.
Plate: TLC silica gel plate R (5 µm). Calculation of percentage contents:
Mobile phase: methanol R, glacial acetic acid R, toluene R - correction factor: multiply the peak area of impurity E by
(5:25:75 V/V/V).
0.2;
Application: 10 µL. - for each impurity, use the concentration of biotin in
Development: over 3/4 of the plate. reference solution (a).
Drying: in a current of warm air. Limits:
Detection: allow to cool and spray with 4-dimethylami- - impurities A, E: for each impurity, maximum 0.5 per cent;
nocinnamaldehyde solution R; examine immediately in - impurity C: maximum 0.2 per cent;
daylight.
- unspecified impurities: for each impurity, maximum
Results: the principal spot in the chromatogram obtained 0.10 per cent;
with the test solution is similar in position and size to the
principal spot in the chromatogram obtained with the - total: maximum 2.0 per cent;
-
1 reference solution.
TESTS
Solution S. Dissolve 0.250 g in a 4 g/L solution of sodium
- reporting threshold: O.OS per cent.
Loss on drying (2.2.32) : maximum 1.0 per cent, determined
on 1.000 g by drying in an oven at 105 ºC.
hydroxide R and dilute to 25.0 mL with the same alkaline 1.0 g.
solution.
ASSAY
Appearance of solution. Solution Sis clear (2.2.1) and
Suspend 0.200 g in 5 mL of dimethylformamide R. Heat
colourless (2.2.2, Method JI).
until the substance has dissolved completely. Add 50 mL
Speci:fic optical rotation (2.2. 7): + 89 to + 93 (dried of ethanol R and titrate with 0.1 M tetrabutylammonium
substance), determined on solution S. hydroxide, determining the end-point potentiometrically
Related substances. Liquid chromatography (2.2.29). Prepare (2.2.20).
the solutions immediately before use and keep protected from 1 mL of 0.1 M tetrabutylammonium hydroxide is equivalent
bright light. to 24.43 mg of C 10H 16 Np 3S.
Biotin EUROPEAN PHARMACOPOEIA 9.5
STORAGE
Store protected from light.
IMPURITIES
Specified impurities: A, C, E.
Other detectable impurities (the following substances would,
if present at a sufficient level, be detected by one or other of
the tests in the monograph. They are limited by the general
acceptance criterion for other/unspecified impurities and/or
by the general monograph Substances for pharmaceutical use
(2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.1 O. Control of
impurities in substances for pharmaceutical use): B, D, F, G, H. E. 5-[ (3aS,4S,6aR)-1-benzyl-2-oxohexahydro-1H-
thieno[3,4-d]imidazol-4-yl]pentanoic acid and
H H H__ ~ ':l H 5-[(3aS,4S,6aR)-3-benzyl-2-oxohexahydro-1H-thieno[3,4-
o=<N~s~N)=o
N~ co H s~N 2
d]imidazol-4-yl]pentanoic acid,
H H H H
A. 5-[ (3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-
d]imidazol-4-yl]-2-[[ (3aS,4S,6aR)-2-oxohexahydro-1H-
thieno [3,4-d] imidazol-4-yl] propyl] pentanoic acid,
H
~t;_.-~C02H F. diethyl 4-[ (3aS,4S,6aR)-l ,3-dibenzyl-2-oxohexahydro-1H-
o=< S C02H
thieno [3,4-d] imidazol-4-yl]butane- l, 1-dicarboxylate,
N
H H
B. 4-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-
d] imidazol-4-yl] bu tane-1, 1-dicarboxylic acid,
H H
--~C02H
H2N··· S
......
1-l-l\J ...
'____,,
H G. (3aR,8aS,8bS)- l ,3-dibenzyl-2-oxodecahydrothie-
no [ l ',2 ': l ,2] thieno [3,4-d] imidazol-5-ium,
C. 5-[ (2S,3S,4R)-3,4-diaminothiolan-2-yl]pentanoic acid,
H H
O
=< ~t··~C0 2 H
N
H H
S H CH3
andepimeratC*
D. (2RS)-2-methyl-5-[(3aS,4S,6aR)-2-oxohexahydro-1H- H. 2-ethyl-5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-
thieno [3,4-d] imidazol-4-yl] pentanoic acid, d] imidazol-4-yl] pentanoic acid.

e
Clomifene citrate.................................................................... 5635 Codeine monohydrate ............................................................ 5639
Codeine hydrochloride dihydrate ......................................... 5636 Codeine phosphate hemihydrate .......................................... 5641

EUROPEAN PHARMACOPOEIA 9.5 Clomifene citrate
o1/2008:0997 System suitability: reference solution (a):

corrected 9.5 - peak-to-valley ratio: minimum lS, where HP = height
above the baseline of the peak due to impurity A and
Hv = height above the baseline of the lowest point of the
curve separating this peak from the peak due to clomifene;
if necessary, adjust the concentration of acetonitrile in the
CLOMIFENE CITRATE mobile phase;
- the chromatogram obtained is similar to the chromatogram
Clomifeni citras supplied with clomifene citrate far performance test CRS.
Limits:
- impurity A: not more than the area of the principal peak
in the chromatogram obtained with reference solution (b)
(2.0 per cent);
- impurities B, C, D, E, F, G, H: for each impurity, not
more than 0.5 times the area of the principal peak in the
chromatogram obtained with reference solution (b) (1.0 per
and (Z)-isomer
cent);
- total: not more than 1.25 times the area of the principal
C 32 H 36 ClN0 8 Mr 598.1 peak in the chromatogram obtained with reference
[50-41-9] solution (b) (2.5 per cent);
DEFINITION - disregard limit: 0.025 times the area of the principal peak
Mixture of the (E)- and (Z)-isomers of2-[4-(2-chloro-l,2-
(O.OS per cent); disregard any peak with a retention time
diphenylethenyl )phenoxy] -N,N-diethylethanamine
relative to the clomifene peak of 0.2 or less.
dihydrogen citrate.
Content: 98.0 per cent to 101.0 per cent (anhydrous substance). (Z)-isomer. Liquid chromatography (2.2.29).
Test solution. Dissolve 25 mg of the substance to be examined
CHARACTERS in 25 mL of 0.1 M hydrochloric acid, add 5 mL of 1 M sodium
Appearance: white or pale yellow, crystalline powder. hydroxide and shake with 3 quantities, each of 25 mL, of
Solubility: slightly soluble in water, sparingly soluble in ethanol-free chloroform R. Wash the combined extracts with
ethanol (96 per cent). 1O mL of water R, dry over anhydrous sodium sulfate R and
dilute to 100 mL with ethanol-free chloroform R. To 20 mL
IDENTIFICATION of this solution add 0.1 mL of triethylamine R and dilute to
-
A. Infrared absorption spectrophotometry (2.2.24).
Comparison: clomifene citrate CRS.

B. Dissolve about 5 mg in 5 mL of a mixture of 1 volume of
acetic anhydride R ;nd 5 volumes of pyridine R, then heat
100 mL with hexane R.
Reference solution. Dissolve 25 mg of clomifene citrate CRS in
25 mL of 0.1 M hydrochloric acid, add 5 mL of 1 M sodium
hydroxide and shake with 3 quantities, each of 25 mL, of
ethanol-free chloroform R. Wash the combined extracts with
in a water-bath. A deep red colour is produced. 10 mL of water R, dry over anhydrous sodium sulfate R and
dilute to 100 mL with ethanol-free chloroform R. To 20 mL
TESTS of this solution add 0.1 mL of triethylamine R and dilute to
100 mL with hexane R.
Prepare the solutions protected from light in brown-glass
vessels. Ensure mínimum exposure of the solutions to daylight Column:
until they are required for chromatography. - size: l = 0.3 m, 0 = 4 mm;
Related substances. Liquid chromatography (2.2.29). - stationary phase: silica gel far chromatography R (10 µm).
Test solution. Dissolve 12.5 mg of the substance to be Mobile phase: triethylamine R, ethanol-free chloroform R,
examined in the mobile phase and dilute to 10.0 mL with the hexane R (1:200:800 V!V!V).
mobile phase. Flow rate: 2 mL/min.
Reference solution (a). Dissolve 12.5 mg of clomifene citrate Detection: spectrophotometer at 302 nm.
f or performance test CRS in the mobile phase and dilute to
10.0 mL with the mobile phase.
Equilibration: with the mobile phase for about 2 h.
Reference solution (b ). Dilute 1.0 mL of the test solution to Injection: 50 µL.
SO.O mL with the mobile phase. Identification of peaks: the chromatogram obtained with the
reference solution shows a peak due to the (E)-isomer just
Column:
before a peak dueto the (Z)-isomer.
- size: l = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped butylsilyl silica gel for
- resolution: minimum 1.0 between the peaks due to the
chromatography R (5 µm).
(E)- and (Z)-isomers; if necessary, adjust the relative
Mobile phase: mix 400 mL of acetonitrile far chromatography R proportions of ethanol-free chloroform and hexane in the
with 600 mL of water for chromatography R and add 8.0 mL mobile phase.
of diethylamine R; adjust to pH 6.2 with about 1-2 mL
Measure the area of the peak dueto the (Z)-isomer in the
of phosphoric acid R, taking care to reduce progressively
chromatograms obtained with the test solution and the
the volume of each addition as the required pH is approached.
reference solution. Calculate the content of the (Z)-isomer, as
Flow rate: 1.2 mL/min. a percentage of the total clomifene citrate present, from the
Detection: spectrophotometer at 233 nm. declared content of clomifene citrate CRS.
Equilibration: with the mobile phase for about 1 h. Limit:
Injection: 10 µL. - (Z)-isomer: 30.0 per cent to SO.O per cent.
Run time: 4 times the retention time of clomifene. Water (2.5.12): maximum 1.0 per cent, determined on 1.000 g.
General Notices (1) apply to all monographs and other texts S63S
Codeine hydrochloride dihydrate EUROPEAN PHARMACOPOEIA 9.5
CI
ASSAY
Dissolve O.SOO g in SO mL of anhydrous acetic acid R. Titrate
with 0.1 M perchloric acid, determining the end-point
potentiometrically (2.2.20).
1 mL of 0.1 M perchloric acid is equivalent to S9.81 mg CI
of C 32 H 36 ClN0 8 •
STORAGE and (Z)-isomer

Protected from light.
F. 2-[ 4-[2-chloro-2-( 4-chlorophenyl)-1-phenylethenyl]-
IMPURITIES phenoxy]-N,N-diethylethanamine,
Specified impurities: A, B, C, D, E, F, G, H.
or (Z)-isomer
and (Z)-isomer
GH. 2-[2-chloro-4-(2-chloro-l ,2-diphenylethenyl)phenoxy]-
A. 2-[4-(l,2-diphenylethenyl)phenoxy]-N,N-diethyl- N,N-diethylethanamine (G. higher-melting-point isomer;
ethanamine, H. lower-melting-point isomer).
07/2018:1412
CODEINE HYDROCHLORIDE
R [4- [2-( diethylamino )ethoxy] phenyl ]phenylmethanone,
DIHYDRATE
Codeini hydrochloridum dihydricum
CH3
1
Al,_ . HCI. 2H20
C. (2RS)-2-[ 4-[2-( diethylamino )ethoxy]phenyl]-1,2- H,Cr::1--i·oH

diphenylethanone,
C 18 H 22 ClN0 3,2H 20
DEFINITION
4,Sa-Epoxy-3-methoxy-17-methyl-7,8-didehydromorphinan-
6a-ol hydrochloride dihydrate.
Content: 99.0 per cent to 101.0 per cent (anhydrous substance).
CHARACTERS
Appearance: white or almost white, crystalline powder or
small, colourless crystals.
Solubility: soluble in water, slightly soluble in ethanol (96 per
cent), practically insoluble in cyclohexane.
D. 2,2-bis[4-[2-(diethylamino)ethoxy]phenyl]-l,2-
diphenylethanone, IDENTIFICATION
First identification: A, D, F.
CI Second identification: B, C, D, E, F.
Comparison: codeine hydrochloride dihydrate CRS.
B. To S mL of solution S (see Tests) add 1 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 R and dry at
and (Z)-isomer 100-lOS ºC. It melts (2.2.15) at lSS ºC to 1S9 ºC.
CI C. To about 10 mg add 1 mL of sulfuric acid R and O.OS mL
of ferric chloride solution R2 and heat on a water-bath. A
E. 2-[4-[1,2-bis(4-chlorophenyl)ethenyl]phenoxy]-N,N- blue colour develops. Add O.OS mL of nitric acid R. The
diethylethanamine, colour changes to red.

EUROPEAN PHARMACOPOEIA 9.S Codeine hydrochloride dihydrate
D. Solution S gives reaction (a) of chlorides (2.3.1). Calculation of percentage contents:

E. It gives the reaction of alkaloids (2.3.1). - correction factors: multiply the peak areas of the following
F. Water (see Tests). impurities by the corresponding correction factor:
impurity C = O. 7; impurity G = 0.2; impurity I = 1.3;
TESTS - for each impurity, use the concentration of codeine in
Solution S. Dissolve 2.00 g in carbon dioxide-free water R, reference solution (a).
prepared from distilled water R, and dilute to SO.O mL with Limits:
the same solvent.
- impurity A: maximum 1.0 per cent;
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than reference solution Y6 (2.2.2, - impurity H: maximum 0.2S per cent;
Method JI). - impurities C, D, E: for each impurity, maximum 0.2 per
Acidity or alkalinity. To S mL of solution S add S mL of cent;
carbon dioxide-free water R. Add O.OS mL of methyl red - impurities B, F, G, I: for each impurity, maximum O.lS per
solution R and 0.2 mL of 0.02 M hydrochloric acid; the solution cent;
is red. Add 0.4 mL of 0.02 M sodium hydroxide; the solution - unspecified impurities: for each impurity, maximum
becomes yellow. 0.10 per cent;
Specific optical rotation (2.2.7): - 117 to - 121 (anhydrous - total: maximum l.S per cent;
substance).
Dilute S.0 mL of solution S to 10.0 mL with water R.
Sulfates (2.4.13) : maximum 0.1 per cent.
Related substances. Liquid chromatography (2.2.29).
Solution A: O.S per cent V/V solution of phosphoric acid R. Dilute S mL of solution S to 20 mL with distilled water R.
Test solution. Dissolve 0.190 g of the substance to be examined Water (2.5.12): 8.0 per cent to 10.S per cent, determined on
in solution A and dilute to SO.O mL with solution A. 0.250 g.
Reference solution (a). Dilute 2.0 mL of the test solution to ASSAY
100.0 mL with solution A. Dilute 1.0 mL of this solution to
10.0 mL with solution A. Dissolve 0.300 gin a mixture of S mL of 0.01 M hydrochloric
acid and 50 mL of ethanol (96 per cent) R. Titrate with
Reference solution (b ). Dissolve 3.0 mg of codeine for system
0.1 M sodium hydroxide, determining the end-point
suitability CRS (containing impurities A, B, C, D, E, F, G, H
potentiometrically (2.2.20). Read the volume added between
and I) in 1.0 mL of solution A.
the 2 points of inflexion.
Column:
1 mL of 0.1 M sodium hydroxide is equivalent to 33.S9 mg
- size: l = 0.07S m, 0 = 3.0 mm; of C 18 H 22 ClN03'
- stationary phase: end-capped octadecylsilyl multi-layered
organosilica polymer R (I.9 µm). STORAGE
- temperature: 40 ºC. Protected from light.
ivíobile phase:
IMPURITIES
- mobile phase A: mix 4 volumes of acetonitrile R and
96 volumes of a 20 g/L solution of glacial acetic acid R Specified impurities: A, B, C, D, E, F, G, H, l.
previously adjusted to pH 4.5 with a SOO g/L solution of Other detectable impurities (the following substances would,
sodium hydroxide R; if present at a sufficient level, be detected by one or other of
- mobile phase B: acetonitrile R; the tests in the monograph. They are limited by the general
Time Mobile phase A Mobile phase B
(min) (per cent V/V) (per cent V/V) (2034). It is therefore not necessary to identify these impurities
o- 5 100 o for demonstration of compliance. See also 5.1 O. Control of
5 - 7.33 100 7 93 077 impurities in substances for pharmaceutical use):], K, L, M.
7.33 - 10.33 93 7 67 7 7 33
10.33 - 12 67 33

Detection: spectrophotometer at 280 nm.
Injection: 3 µL.
Identification of impurities: use the chromatogram supplied
with codeine for system suitability CRS and the chromatogram A. 4,5a-epoxy-3,6a-dimethoxy-17-methyl-7,8-
obtained with reference solution (b) to identify the peaks due didehydromorphinan (methylcodeine),
to impurities A, B, C, D, E, F, G, H and I.
Relative retention with reference to codeine (retention CH 3
time = about 6.0 min): impurity B = about 0.3; 1
~.
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 1.4S;
impurity A = about l.S; impurity G = about 1.6.
System suitability: reference solution (b): HO o· H H OH
- resolution: mínimum 2.5 between the peaks due to

impurities F and H; mínimum l.S between the peaks due B. 4,5a-epoxy-17-methyl-7,8-didehydromorphinan-3,6a-diol
to impurities D and A. (morphine),
General Notices (1) apply to all monographs and other texts S637
Codeine hydrochloride dihydrate EUROPEAN PHARMACOPOEIA 9.5
H. 4,5a-epoxy-3-methoxy-7,8-didehydromorphinan-6a-ol
(norcodeine),
C. 4,5a:4' ,5' a-diepoxy-3,3' -dimethoxy-17,17' -dimethyl-

7,7' ,8,8' -tetradehydro-2,2' -bimorphinan-6a,6' a-diol
(codeine dimer),
CH3
1
H ~l,
HO.~- ~
H O O
I. 4,5a-epoxy-3-methoxy-17-methyl-7,8-didehydro-
morphinan-6-one (codeinone),
·~ .· \_-¡) O, - O/H H····oH
H CH3
N .H
1
CH 3
D. 2-[(4,5a-epoxy-6a-hydroxy-17-methyl-7,8-
didehydromorphinan-3-yl)oxy]-4,5a-epoxy-3-
methoxy-17-methyl-7,8-didehydromorphinan-6a-ol
(3-0-(codein-2-yl)morphine), J. (17RS)-4,5a-epoxy-6a-hydroxy-3-methoxy-17-methyl-7,8-
didehydromorphinan 17-oxide (codeine N-oxide),
CH 3
1
HO H N
H,C~OH K. 4,5a-epoxy-14-hydroxy-3-methoxy-17-methyl-7,8-
E. 4,5a-epoxy-3-methoxy-17-methyl-7,8-didehydro- didehydromorphinan-6-one ( 14-hydroxycodeinone ),
morphinan-6a, 1o~ -diol,
L. 4,5a-epoxy-6-methoxy-17-methyl-6,7,8,14-
tetradehydromorphinan-3-ol (oripavine),
F. 4,5a-epoxy-3-methoxy-17-methyl-7,8-didehydro-
morphinan-6a, 14-diol,
CH 3
1
H N
~O~CH,
H3CO o H OH "'=lV
N H
1
CH3
M. 7,7'-oxybis(4,5a-epoxy-3-methoxy-17-methyl-6,7,8,14-
G. 4,5a-epoxy-3,6-dimethoxy-17 -methyl-6, 7,8,14- tetradehydromorphinan-6-ol) (7,7' -oxybis( 6-0-
tetradehydromorphinan (thebaine), demethylthebaine)).

EUROPEAN PHARMACOPOEIA 9.5 Codeine monohydrate
07/2018:0076 Reference solution (a). Dilute 2.0 mL of the test solution to

10.0 mL with solution A.
CODEINE MONOHYDRATE and I) in 1.0 mL of solution A.
Column:
Codeinum monohydricum - size: l = 0.075 m, 0 = 3.0 mm;
organosilica polymer R (1.9 µm).
- temperature: 40 ºC.
Mobile phase:
- mobile phase A: mix 4 volumes of acetonitrile R and
96 volumes of a 20 g/L solution of glacial acetic acid R
previously adjusted to pH 4.5 with a 500 g/L solution of
sodium hydroxide R;
C18H 21 N0 3,H 20 Mr 317.4
[6059-47-8] - mobile phase B: acetonitrile R;
DEFINITION
(min) (per cent V/V) (per cent V/V)
4,Sa-Epoxy-3-methoxy-17-methyl-7,8-didehydromorphinan- 0-5 100 o
6a-ol monohydrate.
Content: 99.0 per cent to 101.0 per cent (dried substance). 5 - 7.33 100 -7 93 o -7 7
7.33 - 10.33 93 -7 67 7 -7 33
CHARACTERS
Appearance: white or almost white, crystalline powder or 10.33 - 12 67 33
colourless crystals.
Solubility: slightly soluble in water, freely soluble in ethanol
(96 per cent). Detection: spectrophotometer at 280 nm.
Injection: 3 µL.
IDENTIFICATION
First identification: A, C, F. with codeine for system suitability CRS and the chromatogram
Second identification: A, B, D, E, F. obtained with reference solution (b) to identify the peaks due
A. Melting point (2.2.14): l SS ºC to 159 ºC. to impurities A, B, C, D, E, F, G, H and l.
B. Ultraviolet and visible absorption spectrophotometry Relative retention with reference to codeine (retention
(2.2.25). time = about 6.0 min): impurity B = about 0.3;
Test solution. To 2.0 mL of solution S (see Tests) add 50 mL impurity E = about 0.4; impurity F = about 0.8;
of water R then 1O mL of 1 M sodium hydroxide and dilute impurity H = about 0.9; impurity C = about 1.2;
to 100.0 mL with water R. impurity 1 = about 1.4; impurity D = about l .4S;
impurity A = about l .S; impurity G = about 1.6.
Spectral range: 2S0-350 nm.
Absorption maximum: at 284 nm.
Specific absorbance at the absorption maximum: about 50 - resolution: mínimum 2.5 between the peaks due to
(dried substance). impurities F and H; mínimum 1.S between the peaks due
-
C. Infrared absorption spectrophotometry (2.2.24).
Comparison: codeine CRS.

D. To about 10 mg add 1 mL of sulfuric acid R and O.OS mL
of ferric chloride solution R2 and heat on a water-bath. A
to impurities D and A.
- correction factors: multiply the peak areas of the following
impurities by the corresponding correction factor:
impurity C = O. 7; impurity G = 0.2; impurity 1 = 1.3;
blue colour develops. Add 0.05 mL of nitric acid R. The - for each impurity, use the concentration of codeine in
colour changes to red. reference solution (a).
E. It gives the reaction of alkaloids (2.3.1). Limits:
F. Loss on drying (see Tests). - impurity A: maximum 1.0 per cent;
TESTS - impurity H: maximum 0.2S per cent;
Solution S. Dissolve SO mg in water R and dilute to 1O.O mL - impurities C, D, E: for each impurity, maximum 0.2 per
with the same solvent. cent;
Appearance of solution. Solution S is clear (2.2.1) and - impurities B, F, G, J: for each impurity, maximum O.lS per
colourless (2.2.2, Method JI). cent;
Specific optical rotation (2.2.7): - 142 to - 146 (dried - unspecified impurities: for each impurity, maximum
substance). 0.10 per cent;
Dissolve O.SO gin ethanol (96 per cent) R and dilute to 2S.O mL - total: maximum 1.5 per cent;
with the same solvent. - reporting threshold: O.OS per cent.
Related substances. Liquid chromatography (2.2.29). Loss on drying (2.2.32): 4.0 per cent to 6.0 per cent,
Solution A: 0.5 per cent V/V solution of phosphoric acid R. determined on 1.000 g by drying in an oven at 105 ºC.
Test solution. Dissolve 0.160 g of the substance to be examined Sulfated ash (2.4.14) : maximum 0.1 per cent, determined on
in solution A and dilute to 50.0 mL with solution A. 1.0 g.
Codeine monohydrate EUROPEAN PHARMACOPOEIA 9.5
ASSAY
Dissolve 0.250 g in 50 mL of anhydrous acetic acid R. Titrate
1 mL of 0.1 M perchloric acid is equivalent to 29.94 mg
of C 18 H 21 N0 3•
STORAGE
E. 4,5a-epoxy-3-methoxy-17 -methyl- 7,8-didehydro-
Protected from light. morphinan-6a, 1o~ -diol,
IMPURITIES
Specified impurities: A, B, C, D, E, F, G, H, l.
for demonstration of compliance. See also 5.1 O. Control of F. 4,5a-epoxy-3-methoxy-17-methyl-7,8-didehydro-
impurities in substances for pharmaceutical use): J, K, L, M. morphinan-6a, 14-diol,
CH3
1
H N
H,C~OCH3
A. 4,Sa-epoxy-3,6a-dimethoxy-17-methyl-7,8- G. 4,5a-epoxy-3,6-dimethoxy-17 -methyl-6, 7,8,14-
didehydromorphinan (methylcodeine ), tetradehydromorphinan (thebaine),
H
CH,
H,C~OH
1 v
H N
H~OH
B. 4,5a-epoxy-17-methyl-7 ,8-didehydromorphinan-3,6a-diol
H. 4,5a-epo:xy-3-methoxy-7 ,8-didehydromorphinan-6a-ol
(norcodeine),
(morphine),
N H I. 4,Sa-epoxy-3-methoxy-17-methyl-7,8-didehydro-
1 morphinan-6-one (codeinone ),
CH 3
C. 4,Sa:4' ,5 'a-diepo:xy-3,3' -dimethoxy-17, 17' -dimethyl-

7, 7' ,8,8' -tetradehydro-2,2' -bimorphinan-6a,6' a-diol
(codeine dimer),
J. (17RS)-4,5a-epoxy-6a-hydroxy-3-methoxy-17-methyl-7,8-
didehydromorphinan 17-oxide (codeine N-oxide ),
D. 2-[(4,5a-epoxy-6a-hydroxy-17-methyl-7,8-
didehydromorphinan-3-yl)oxy]-4,Sa-epoxy-3-
methoxy-17 -methyl-7,8-didehydromorphinan-6a-ol K. 4,5a-epoxy-14-hydroxy-3-methoxy-17 -methyl-7,8-
(3-0-(codein-2-yl)morphine), didehydromorphinan -6-one ( 14-hydroxycodeinone),

EUROPEAN PHARMACOPOEIA 9.5 Codeine phosphate hemihydrate
B. Infrared absorption spectrophotometry (2.2.24).

Preparation: dissolve 0.20 g in 4 mL of water R. Add 1 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 R and dry at 100-105 ºC.
L. 4,5a-epoxy-6-methoxy-17-methyl-6,7,8,14- Comparison: codeine CRS.
tetradehydromorphinan-3-ol (oripavine), C. Dissolve 0.20 gin 4 mL of water R. Add 1 mL of a mixture
CH 3 of equal volumes of strong sodium hydroxide solution R
1
and water R and initiate crystallisation, if necessary, by
~o
u-Al, ~HOf'
··º
" ..
-- ~
OCH,
¡)
scratching the wall of the tube with a glass rod and cooling
in iced water. The precipitate, washed with water R and
dried at 100-105 ºC, melts (2.2.14) at 155 ºC to 159 ºC.
D. To about 10 mg add 1 mL of sulfuric acid R and 0.05 mL
H3CO O H OH \ of ferric chloride solution R2 and heat on a water-bath. A
N H blue colour develops. Add 0.05 mL of nitric acid R. The
1
CH 3 colour changes to red.
E. Loss on drying (see Tests).
M. 7,7'-oxybis(4,5a-epoxy-3-methoxy-17-methyl-6,7,8,l4- F. Solution S gives reaction (a) of phosphates (2.3.1).
tetradehydromorphinan-6-ol) (7, 7' -oxybis( 6-0-
demethylthebaine)). G. It gives the reaction of alkaloids (2. 3.1).
TESTS
07/2018:0074 Solution S. Dissolve 1.00 gin carbon dioxide-free water R
prepared from distilled water R and dilute to 25.0 mL with
the same solvent.
pH (2.2.3): 4.0 to 5.0 for solution S.
CODEINE PHOSPHATE Specific optical rotation (2.2. 7) : - 98 to - 102 (dried
substance).
HEMIHYDRATE
Dilute 5.0 mL of solution S to 10.0 mL with water R.
Codeini phosphas hemihydricus Related substances. Liquid chromatography (2.2.29).
Solution A: 0.5 per cent V/V solution of phosphoric acid R.
Test solution. Dissolve 0.190 g of the substance to be examined
in solution A and dilute to SO.O mL with solution A.
10.0 mL with solution A.
and I) in 1.0 mL of solution A.
Column:
C 18 H 24N0 7P, 1hH2 0 Mr 406.4
[41444-62-6] size: l = 0.075 m, 0 = 3.0 mm;
DEFINITION organosilica polymer R (1.9 µm).
4,Sa-Epoxy-3-methoxy-17-methyl-7,8-didehydromorphinan- - temperature: 40 ºC.
6a-ol phosphate hemihydrate. Mobile phase:
Content: 99.0 per cent to 101.0 per cent (dried substance). - mobile phase A: mix 4 volumes of acetonitrile R and
CHARACTERS 96 volumes of a 20 g/L solution of glacial acetic acid R
previously adjusted to pH 4.5 with a 500 g/L solution of
Appearance: white or almost white, crystalline powder or
sodium hydroxide R;
small, colourless crystals.
- mobile phase B: acetonitrile R;
Solubility: freely soluble in water, slightly soluble or very
slightly soluble in ethanol (96 per cent). Time Mobile phase A Mobile phase B
IDENTIFICATION o-5 100 o
First identification: B, E, F.
5 - 7.33 100 -7 93 o -7 7
Second identification: A, C, D, E, F, G.
A. Ultraviolet and visible absorption spectrophotometry 7.33 - 10.33 93 -7 67 7 -7 33
(2.2.25). 10.33 - 12 67 33
Test solution. Dilute 1.0 mL of solution S (see Tests) to
100.0 mL with water R. To 25.0 mL of this solution add Flow rate: 1.0 mL/min.
25 mL of water R then 10 mL of 1 M sodium hydroxide and Detection: spectrophotometer at 280 nm.
dilute to 100.0 mL with water R. Injection: 3 µL.
Spectral range: 250-350 nm. Identification of impurities: use the chromatogram supplied
Absorption maximum: at 284 nm. with codeine for system suitability CRS and the chromatogram
Specific absorbance at the absorption maximum: about 38 obtained with reference solution (b) to identify the peaks due
(dried substance). to impurities A, B, C, D, E, F, G, H and I.
Codeine phosphate hemihydrate EUROPEAN PHARMACOPOEIA 9.5
CH 3
Relative retention with reference to codeine (retention 1
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 1.5; impurity G = about 1.6.
HO o· H H OH
- resolution: minimum 2.5 between the peaks due to B. 4,Sa-epoxy-17-methyl-7,8-didehydromorphinan-3,6a-diol
impurities F and H; minimum 1.5 between the peaks due (morphine),
to impurities D and A.
correction f actors: multiply the peak areas of the following
impurity C = O. 7; impurity G = 0.2; impurity I = 1.3;
for each impurity, use the concentration of codeine in
reference solution (a).
N H
Limits: 1
CH 3
- impurity A: maximum 1.0 per cent;
C. 4,Sa:4',S'a-diepoxy-3,3'-dimethoxy-17,17'-dimethyl-
- impurity H: maximum 0.25 per cent; 7,7',8,8'-tetradehydro-2,2'-bimorphinan-6a,6'a-diol
- impurities C, D, E: for each impurity, maximum 0.2 per (codeine dimer),
cent;
impurities B, F, G, I: for each impurity, maximum 0.15 per
cent;
0.10 per cent;
- total: maximum 1.5 per cent;
Sulfates (2.4.13): rnaximum 0.1 per cent
Dilute 5 mL of solution S to 20 mL with distilled water R.
Loss on drying (2.2.32): 1.5 per cent to 3.0 per cent,
determined on 1.000 g by drying in an oven at 105 ºC. D. 2-[(4,Sa-epoxy-6a-hydroxy-17-methyl-7,8-
didehydromorphinan -3-yl) oxy] -4,Sa-epoxy-3-
ASSAY methoxy-17 -methyl-7,8-didehydromorphinan-6a-ol
(3-0-(codein-2-yl)morphine),
Dissolve 0.350 g in 50 mL of anhydrous acetic acid R. Titrate
of C 18 H 24 N0 7 P.
STORAGE
Protected from light.
E. 4,Sa-epoxy-3-methoxy-17-methyl-7,8-didehydro-
IMPURITIES morphinan-6a, 1o~ -diol,
Specified impurities: A, B, C, D, E, F, G, H, I.
impurities in substances for pharmaceutical use): J, K, L, M. F. 4,Sa-epoxy-3-methoxy-17-methyl-7,8-didehydro-
morphinan-6a, 14-diol,
A. 4,Sa-epoxy-3,6a-dimethoxy-17-methyl-7,8- G. 4,Sa-epoxy-3,6-dimethoxy-17-methyl-6,7,8,14-
didehydromorphinan (methylcodeine), tetradehydromorphinan (thebaine),

EUROPEAN PHARMACOPOEIA 9.5 Codeine phosphate hemihydrate
H
H N
H,co~º"
H. 4,5a-epo:xy-3-metho:xy-7,8-didehydromorphinan-6a-ol K. 4,5a-epo:xy-14-hydro:xy-3-methoxy-17-methyl-7,8-
(norcodeine), didehydromorphinan-6-one ( 14-hydroxycodeinone),
CH 3
1
HO
~ o·· H OCH3
L. 4,5a-epo:xy-6-methoxy-17-methyl-6,7,8,l 4-
I. 4,5a-epo:xy-3-metho:xy-17-methyl-7,8-didehydro- tetradehydromorphinan -3-ol (oripavine),
morphinan-6-one (codeinone), ?H3
rA\\ o~"ºr
~
".
--
··º OCH,
\j
H3CO O H OH \
N H
1
CH 3
M. 7,7' -o:xybis( 4,5a-epo:xy-3-metho:xy-17-methyl-6,7,8,14-

J. (17RS)-4,5a-epoxy-6a-hydro:xy-3-metho:xy-17-methyl-7,8- tetradehydromorphinan-6-ol) (7, 7' -o:xybis( 6-0-
didehydromorphinan 17-oxide (codeine N-oxide), demethylthebaine)).

D
Deferiprone ............................................................................. 564 7

EUROPEAN PHARMACOPOEIA 9.S Deferiprone
07/2018:2236 Mobile phase: acetonitrile R, solution A (10:90 V/V).

Preconditioning of the column: rinse for 20 min with the
mobile phase before each series of injections.
DEFERIPRONE Injection: 20 µL of test solution (a) and reference solutions (a)
and (b).
Deferipronum Run time: 3.S times the retention time of deferiprone.
Identification of impurities: use the chromatogram obtained
with reference solution (a) to identify the peak dueto
impurity B.
Relative retention with reference to deferiprone (retention
time = about 12 min): impurity B = about O.S.
C7 H 9N0 2 System suitability: reference solution (a):
[30652-11-0] - resolution: minimum S.O between the peaks due to
DEFINITION impurity B and deferiprone.
3-Hydroxy-l,2-dimethylpyridin-4-(lH)-one. Calculation of percentage contents:
Content: 98.0 per cent to 102.0 per cent (anhydrous substance). - for each impurity, use the concentration of deferiprone in
reference solution (b).
CHARACTERS Limits:
Appearance: white or pinkish-white powder. - impurity B: maximum 0.10 per cent;
Solubility: sparingly soluble in water, slightly soluble in - unspecified impurities: for each impurity, maximum
anhydrous ethanol, practically insoluble in heptane. O.OS per cent;
IDENTIFICATION - total: maximum 0.2 per cent;
Infrared absorption spectrophotometry (2.2.24). - reporting threshold: 0.03 per cent.
Comparison: deferiprone CRS. Water (2.5.32): maximum O.S per cent, determined on 0.100 g
by direct sample introduction.
TESTS
Related substances. Liquid chromatography (2.2.29). Use 1.0 g.
only colourless glassware. Protect the solutions from light.
Solution A. Dissolve 2.91 g of sodium edetate R, 4.01 g of sodium ASSAY
octanesulfonate monohydrate R and 6.20 g of dipotassium Liquid chromatography (2.2.29) as described in the test for
hydrogen phosphate R in water for chromatography R and related substances with the following modifications.
dilute to 2000 mL with the same solvent; adjust to pH 3.0 with Mobile phase: acetonitrile R, solution B (10:90 V/V).
phosphoric acid R. lnjection: 10 µL of test solution (b) and reference solution (c).
Soiution B. Dissoive 0.73 g of sodium edetate R, 1.0 g of sodium Run time: twice the retention time of deferiprone.
octanesulfonate monohydrate R and 1.SS g of dipotassium Retention time: deferiprone = about 7.7 min.
hydrogen phosphate R in water for chromatography R and
dilute to 2000 mL with the same solvent; adjust to pH 3.0 with Calculate the percentage content of C 7 H 9N0 2 taking into
phosphoric acid R. account the assigned content of deferiprone CRS.
Solvent mixture: acetonitrile R, water R (10:90 V/V). IMPURITIES
Test solution (a). Dissolve 0.100 g of the substance to be Specified impurities: B.
examined in a volume of the mobile phase corresponding to Other detectable impurities (the following substances would,
about 2/3 of the final volume and dilute to 100.0 mL with the if present at a sufficient level, be detected by one or other of
mobile phase. the tests in the monograph. They are limited by the general
Test solution (b). Dissolve SO.O mg of the substance to be acceptance criterion for other/unspecified impurities. It
examined in a volume of the solvent mixture corresponding to is therefore not necessary to identify these impurities for
about 2/3 of the final volume and dilute to SO.O mL with the demonstration of compliance. See also 5.1 O. Control of
solvent mixture. Dilute S.O mL of the solution to 200.0 mL impurities in substances for pharmaceutical use): A, C.
with the mobile phase. o
Reference solution (a). Dissolve 2 mg of maltol R (impurity B)
in the mobile phase and dilute to 100.0 mL with the mobile H3c-NA -~,
phase. To 2.5 mL of the solution add 10 mL oftest solution (a)
\_f CH3
and dilute to 100.0 mL with the mobile phase.

A. 1-methyl-3-(methylamino )-l,5-dihydro-2H-pyrrol-2-one,
Reference solution (b ). Dilute 1.0 mL of test solution (a) to
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
to 20.0 mL with the mobile phase.
Reference solution (e). Dissolve SO.O mg of deferiprone CRS
in a volume of the solvent mixture corresponding to about
2/3 of the final volume and dilute to SO.O mL with the solvent B. 3-hydroxy-2-methyl-4H-pyran-4-one (maltol),
mixture. Dilute S.O mL of the solution to 200.0 mL with the
mobile phase.
Column:
- size: l = O.lS m, 0 = 4.6 mm;
- stationary phase: styrene-divinylbenzene copolymer R
(S µm); C. l ,2-dimethyl-4- [(3)-methylimino ]-1,4-dihydropyridin-3-
- temperature: 30 ºC. ol.

E
Estriol. ...................................................................................... 5651 Etanercept ................................................................................ 5652

EUROPEAN PHARMACOPOEIA 9.5 Estriol
07/2018:1203 Time Mobile phase A Mobile phase B

0-9 100 o
9 - 23 100 7 57 o7 43
ESTRIOL 23 - 28 57 43

Estriolum Detection: spectrophotometer at 220 nm.
lnjection: 10 µL of test solution (a) and reference solutions (a)
and (b).
ldentification of impurities: use the chromatogram supplied
with estrío! for system suitability CRS and the chromatogram
obtained with reference solution (a) to identify the peaks due
to impurities A, D, E and F.
C1sH2403 Relative retention with reference to estriol (retention
[50-27-1] time = about 11 min): impurity A = about 0.9S;
impurity F = about 1.4S; ímpurity E = about l.S;
DEFINITION impurity D = about 1.8.
Estra-l,3,5(10)-triene-3,16a, 17~-triol. System suitability: reference solution (a):
Content: 97.5 per cent to 102.0 per cent (dried substance). - peak-to-valley ratio: minimum 5.0, where Hp = height
CHARACTERS above the baselíne of the peak due to ímpuríty A and
Appearance: white or almost white, crystalline powder. curve separating this peak from the peak due to estriol.
-
Solubility: practically insoluble in water, sparingly soluble in Calculation of percentage contents:
ethanol (96 per cent).
- correction factor: multiply the peak area of impurity A by
0.5;
IDENTIFICATION - for each impurity, use the concentration of estríol in
A. Infrared absorption spectrophotometry (2.2.24). reference solution (b ).
Comparison: estriol CRS. Limits:
B. Examine the chromatograms obtained in the assay. - impurity F: maximum 0.5 per cent;
Results: the principal peak in the chromatogram obtained - impurity E: maximum 0.3 per cent;
with test solution (b) is similar in retention time and size - impurities A, D: for each impurity, maximum 0.2 per cent;
to the principal peak in the chromatogram obtained with
reference solution (c).
0.10 per cent;
TESTS - total: maximum 1.0 per cent;
Specific optical rotation (2.2.7): + 60 to + 65 (dried - reporting threshold: O.OS per cent.
substance). Loss on drying (2.2.32): maximum 0.5 per cent, determined
Dissolve 80 mg in anhydrous ethanol R and dilute to 10.0 mL on 1.000 g by drying in an oven at lOS ºC for 3 h.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
Solvent mixture: methanol R, water R (50:50 V/V). related substances with the following modifications.
Test solution (a). Dissolve 25.0 mg of the substance to be Mobile phase:
examined in 25 mL of methanol R and dilute to SO.O mL with
water R. Time Mobile phase A Mobile phase B
o- 5 90 10
Reference solution (a). Dissolve 5 mg of estriol for system 5 - 5.5 90 7 30 10 7 70
suitability CRS (containing impurities A, D, E and F) in 5 mL 5.5 - 7.5 30 70
of methanol R and dilute to 10.0 mL with water R.
Reference solution (b ). Dilute 1.0 mL of test solution (a) to lnjection: S µL of test solution (b) and reference solutions (a)
100.0 mL with the solvent mixture. Dilute 1.0 mL of this and (c).
solution to 10.0 mL with the solvent mixture. Jdentification of impurities: use the chromatogram obtained
Reference solution (e). Dissolve 25.0 mg of estriol CRS in 2S mL with reference solution (a) to identify the peak due to
of methanol R and dilute to SO.O mL with water R. Dilute impurity A.
1.0 mL of the solution to 10.0 mL with the solvent mixture. Relative retention with reference to estriol (retention
Column: time= about 4 min): impurity A= about 0.9.
- size: l = 0.10 m, 0 = 2.1 mm; System suitability: reference solution (a):
stationary phase: end-capped, charged surface, - peak-to-valley ratio: minimum 5.0, where HP = height
ethylene-bridged octadecylsilyl silica gel for chromatography above the baseline of the peak due to impurity A and
(hybrid material) R (l.7 µm); Hv = height above the baseline of the lowest point of the
- temperature: SO ºC. curve separating this peak from the peak due to estriol.
Calculate the percentage content of C 18 H 24 0 3 taking into
Mobile phase:
account the assigned content of estriol CRS.
- mobile phase A: methanol Rl, water for chromatography R
(28:72 V/V); IMPURITIES
- mobile phase B: acetonitrile for chromatography R; Specified impurities: A, D, E, F.
General Notices (1) apply to all monographs and other texts S6Sl
Etanercept EUROPEAN PHARMACOPOEIA 9.5

if present at a suf:ficient level, be detected by one or other of
acceptance criterion for other/unspecifi.ed impurities and/or
by the general monograph Substances for pharmaceutical
use (2034). It is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.1 O. l. 3-hydroxy-17-oxa-17a-homoestra-l,3,5(10)-trien-17a-one,
Control of impurities in substances for pharmaceutical use):
B, C, G, H, I, J, K.
J. estra-1,3,5(10)-triene-3,l7a-diol (17-epi-estradiol),
A. estra-l,3,5(10),9(11)-tetraene-3,16a,l 7p-triol CH3 O
(9,11-didehydroestriol),
H3 JODB13
K. 17-oxoestra-1,3,5(10)-trien-3-yl acetate (estrone acetate).
B. 3-hydroxyestra-1,3,5(10)-trien-17-one (estrone),
07/2018:2895
ETANERCEPT
C. 3-methoxvestra-l,3,5(10)-triene-16a,I 7B-diol (estriol
3-methyl éther), . · . Etanerceptum
LPAQVAFTPY APEPGSTCRL REYYDQTAQM CCSKCSPGQH 40
AKVFCTKTSD TVCDSCEDST YTQLWNWVPE CLSCGSRCSS 80
DQVETQACTR EQNRICTCRP GWYCALSKQE GCRLCAPLRK 120
CRPGFGVARP GTETSDVVCK PCAPGTFSNT TSSTDICRPH 160
QICNVVAIPG NASMDAVCTS TSPTRSMAPG AVHLPQPVST 200
RSQHTQPTPE PSTAPSTSFL LPMGPSPPAE GSTGDEPKSC 240
D. estra-l,3,5(10)-triene-3,l 7P-diol (estradiol), DKTHTCPPCP APELLGGPSV FLFPPKPKDT LMISRTPEVT 280
CVVVDVSHED PEVKFNWYVD GVEVHNAKTK PREEQYNSTY 320
RVVSVLTVLH QDWLNGKEYK CKVSNKALPA PIEKTISKAK 360
GQPREPQVYT LPPSREEMTK NQVSLTCLVK GFYPSDIAVE 400
WESNGQPENN YKTTPPVLDS DGSFFLYSKL TVDKSRWQQG 440
NVFSCSVMHE ALHNHYTQKS LSLSPGK 467
disulfide bridges (the list is not exhaustive):

240-240', 246-246', 249-249', 281-341, 387-445, 281'-341', 387'-445'
E. estra-l,3,5(10)-triene-3,16a,l 7a-triol (17-epi-estriol), N-glycosylation sites:
149, 171, 317, 149', 171', 317'
predominant 0-glycosylation sites:
184, 199, 200, 205, 208, 212, 213, 216, 217, 226, 184', 199', 200', 205',
208', 212', 213', 216', 217', 226'
lVIr approx. 51 200

F. estra-l,3,5(10)-triene-3,16P,I 7p-triol (16-epi-estriol), (monomer without glycosylation)
[185243-69-0]
DEFINITION
Dimeric fusion protein consisting of the extracellular
ligand-binding portion of the human tumour necrosis factor
receptor (TNFR) linked to the Fe portion of human IgG l. The
G. estra-l,3,5(10)-triene-3,16P,l 7a-triol (16,17-epi-estriol), Fe component of etanercept contains the CH 2 domain, the
CH 3 domain and the hinge region, but not the CH 1 domain
of IgG 1. Etanercept consists of 934 amino acids and has an
apparent molecular mass of approximately 150 kDa.
Etanercept represents a glycosylated protein with multiple N-
and 0-linked glycosylation sites. It shows foil occupancy of
N-linked glycans at N149, Nl71 and N317.
It contains one or more suitable buffering and/or stabilising
H. 3,16a-dihydroxyestra-l,3,5(10)-trien-17-one, agents.

EUROPEAN PHARMACOPOEIA 9.5 Etanercept
Content (milligrams of protein per millilitre): as approved by Mobile phase:

the competent authority. - mobile phase A: mix 9.8 mL of anhydrous formic acid R and
Potency: 1.0 x 106 to 2.9 x 106 IU per milligram of protein. 500 mL of water for chromatography R, adjust to pH 4.0
with ammonia R and dilute to 1000 mL with water for
chromatography R;
PRODUCTION
- mobile phase B: acetonitrile R;
Etanercept is produced in a suitable mammalian cell
expression system by a method based on recombinant
DNA (rDNA) technology. During the course of product (min) (per cent V/V) (per cent V/V)
development, it must be demonstrated that the manufacturing o- 2 20 -7 30 80 -7 70
process consistently produces a product with the expected 2 - 67.0 30 -7 52 70 -7 48
0-glycan occupancy using a suitably qualified assay.
67.0 - 67.l 52 -7 80 48 -7 20
Prior to release, the following tests are carried out on each batch
of the final bulk product, unless exemption has been granted by 67.l - 73.0 80 20
the competent authority.
Host-cell-derived proteins. The limit is approved by the
Detection: fluorimeter at 330 nm for excitation and 420 nm
competent authority.
for emission.
Host-cell and vector-derived DNA. The limit is approved Autosampler: set at 2-8 ºC.
by the competent authority.
Injection: 10 µL.
N-Glycan analysis. Use a suitable method developed Identification of peaks: use the chromatogram shown in
according to general chapter 2.2.59. Glycan analysis of Figure 2895.-1 to identify the 2 groups of oligosaccharides
glycoproteins, section 2-3: corresponding to neutral (peaks 1 to 5) and sialylated (peaks
- release the glycans using one of the agents described 6 to 9) N-glycans; record the retention time of each peak in
in Table 2.2.59.-1, for example peptide N-glycosidase F both groups.
(PNGase F); System suitability:
- label the released glycans with one of the fluorescent - the chromatogram obtained with reference solution (a) is
labelling agents described in Table 2.2.59.-2, for example qualitatively similar to the chromatogram supplied with
2-aminobenzamide; etanercept CRS and peaks 1 to 9 are clearly visible;
analyse the labelled glycans by liquid chromatography - no significant peaks are observed in the chromatogram
(2.2.29) using fluorescence detection. obtained with the blank solution.
Results:
The following procedure is given as an example.
- the profile of the chromatogram obtained with the test
Test solution. To 4 µL of the preparation to be examined solution corresponds to that of the chromatogram obtained
(about 25 mg/mL) add 21 µL of water R, 3 µL of 0.25 M with reference solution (b);
sodium phosphate buffer solution pH 7.5 R and 2 µL of a - the retention times of the peaks in the chromatogram
500 000 UímL soiution of peptide N-glycosidase FR. Mix and obtained with the test solution correspond to those in the
incubate at 37 ºC for 20-24 h. Label the released glycans with chromatogram obtained with reference solution (b);
2-aminobenzamide using a suitable procedure. The procedure
employs a combination of reagents optimised and validated - no additional peaks are observed in the chromatogram
for the efficient labelling of glycans, and for the subsequent obtained with the test solution in comparison with the
extraction and recovery of the labelled glycans from the chromatogram obtained with reference solution (b).
reaction. Resuspend or dilute the labelled glycans in 100 µL Calculate the relative peak areas of the individual peaks
of water R. corresponding to neutral and sialylated N-glycans with
reference to the sum of the are as of all retained glycan peaks.
Reference solution (a). Dissolve the contents of a vial of
etanercept CRS in water R to obtain a concentration of about Calculate the percentage contents of the neutral and sialylated
25 mg/mL. Carry out the release and labelling of glycans in groups, using the following expressions:
the same manner as for the test solution. Resuspend or dilute A
the labelled glycans in 100 µL of water R. A+B X 100
Reference solution (b). Use a suitable etanercept in-house B
reference preparation shown to be representative of batches A+ B x 100
tested clinically and batches used to demonstrate consistency
of production. Dilute, if necessary, with water R to obtain a A sum of the areas of the peaks due to neutral
concentration of about 25 mg/mL. Carry out the release and N-glycans;
labelling of glycans in the same manner as for the test solution.
Resuspend or dilute the labelled glycans in 100 µL of water R. B sum of the areas of the peaks due to sialylated
N-glycans.
Blank solution. Prepare at the same time and in the same
manner as for the test solution but using water R instead of NOTE: peaks 2 and 3 are separate peaks, but they are integrated
the preparation to be examined. together.
Limits:
Analyse the labelled glycans by liquid chromatography
(2.2.29). - percentage of neutral N-glycans: as approved by the
competent authority;
Column:
- percentage of sialylated N-glycans: as approved by the
- size: l = 0.25 m, 0 = 4.6 mm; competent authority.
- stationary phase: an amide derivative of silica gel for CHARACTERS
Appearance: clear, almost colourless, slightly yellow or slightly
- temperature: 35 ºC. brown liquid.
2+3
9
8
4
1 1 1 1 1 1 1 1 1 ! 1 1
1 1 1 1 1 1 1 1 1 1 1
30 35 40 45 50 55 60 65 70 75 80 min
Peak Charged Structure Peak Charged Structure Peak Charged Structure Peak Charged Structure
l. No Asialo-, agalacto-, biantennary, 4. No Asialo-, galactosylated 6. Yes Monosialylated-, galactosylated 8. Yes Disialylated, galactosylated
core-fucosylated biantennary biantennary biantennary
2+3. No Asialo-, mono-galactosylated 5. No Asialo-, galactosylated 7. Yes Monosialylated-, galactosylated 9. Yes Disialylated-, galactosylated
biantennary, core-fucosylated biantennary, core-fucosylated biantennary, core-fucosylated biantennary, core-fucosylated
Figure 2895.-1. - Chromatogram for N-glycan analysis of etanercept
IDENTIFICATION CHROMATOGRAPHIC SEPARATION. Liquid

chromatography (2.2.29).
A. It complies with the limits of the assay (potency).
Column:
B. Peptide mapping (2.2.55). - size: l = 0.25 m, 0 = 3.2 mm;
SELECTIVE CLEA VAGE OF THE PEPTIDE BONDS - stationary phase: end-capped octadecylsilyl silica gel for
chromatography R (5 µm) with a pore size of 10 nm;
Test solution. Dilute the preparation to be examined with - temperature: 30-35 ºC.
water R to obtain a concentration of about 15 mg/mL.
Mobile phase:
Reference solution. Dissolve the contents of a vial of - mobile phase A: mix 3 g of trifluoroacetic acid R and
etanercept CRS in water R to obtain a concentration of 2000 mL of water for chromatography R;
about 15 mg/mL. - mobile phase B: mix 2 g of trifluoroacetic acid R,
Reduction and alkylation. To 200 µL of the test solution add 330 mL of water for chromatography R and 1320 mL of
500 µL of guanidine-tris(hydroxymethyl)aminomethane acetonitrile for chromatography R;
buffer solution pH 8.3 R and 7 µL of a 154 g/L solution of Time Mobile phase A Mobile phase B
dithiothreitol R. Mix and incubate at 65 ºC for 15 min. Cool (min) (per cent V/V) (per cent V/V)
in an ice-bath for 5-10 min, then add 15.4 µL of a freshly o- 5 98 2
prepared 185 g/L solution of iodoacetamide R. Mix and
allow to stand protected from light for 1O min. Add 1.4 µL 5 - 125 98 7 50 2 7 50
of a 154 g/L solution of dithiothreitol R. Mix and allow to 125 - 140 50 7 5 50 7 95
stand protected from light for 1O min.
Digestion. To 97 µL of the reduced test solution prepared
previously, add 903 µL of 0.1 M tris-hydrochloride buffer Detection: spectrophotometer at 220 nm.
solution pH 7.5 R. Add 9.6 µL of a 500 000 U/mL solution Autosampler: set at 2-8 ºC.
of peptide N-glycosidase F R and incubate at 37 ºC for 1 h. Injection: 200 µL.
Add 40 µL of a 1 mg/mL solution of trypsin for peptide System suitability:
mapping R and incubate at 37 ºC for 5 h. Heat at 95 ºC for - the chromatogram obtained with the reference solution
5 min and cool in ice for 5 min. Adjust to pH 2 with about is qualitatively similar to the chromatogram supplied
30 µL of a 150 g/L solution of trifluoroacetic acid R. with etanercept CRS.
Carry out the reduction/alkylation and digestion steps for Results: the profile of the chromatogram obtained with
the reference solution in the same manner as for the test the test solution corresponds to that of the chromatogram
solution. obtained with the reference solution.

TESTS System suitability:

pH (2.2.3). As approved by the competent authority. - the peak due to sialic acid in the chromatogram obtained
with reference solution (a) is visible and is similar to the
Sialic acid. Use a suitable method developed according
corresponding peak in the chromatogram supplied with
to general chapter 2.2.59. Glycan analysis of glycoproteins,
etanercept CRS;
section 2-4:
- repeatability: maximum relative standard deviation of
- release the sialic acid by acid hydrolysis pre-treatment of 15 per cent for the sialic acid content expressed as the
the preparation to be examined; molar ratio, determined on 3 consecutive injections of
- label the released sialic acid with a fluorescent reference solution (a);
labelling reagent, for example l,2-diamino-4,5- - the coefficient of determination (R 2 ) calculated for the
methylenedioxybenzene, using a suitable procedure; standard curve is not less than 0.9995.
- analyse the labelled sialic acid by liquid chromatography Results:
(2.2.29) using fluorescence detection.
- the profile of the chromatogram obtained with the test
The following method has been found suitable. solution corresponds to that of the chromatogram obtained
Solution A. Dissolve 8.71 g of arginine R in 40 mL of water R. with reference solution (a).
Add 0.5 mL of a 10 g/L solution of polysorbate 80 R, mix and Limit:
adjust to pH 7.3 with phosphoric acid R. Dilute to 250 mL
with water R. - 8 to 19 moles of sialic acid per mole of etanercept.
Test solution. Dilute the preparation to be examined with Related proteins. Liquid chromatography (2.2.29) : use the
water R to obtain a concentration of about 5 mg/mL. normalisation procedure.
Further dilute with solution A to obtain a concentration Test solution. Dilute the preparation to be examined with
of about 1 mg/mL. To 50 µL of this solution add 50 µL of water R to obtain a concentration of about 2 mg/mL.
a 480 g/L solution of glacial acetic acid R and incubate at Reference solution. Dissolve the contents of a vial of
90 ºC for 65 min. Cool, briefly centrifuge and evaporate etanercept CRS in water R to obtain a concentration of about
to dryness. Label the released sialic acid using a suitable 2 mg/mL.
procedure; for example, add 15 µL of a 1.6 g/L solution of
l,2-diamino-4,5-methylenedioxybenzene dihydrochloride R Column:
containing 78.1 g/L of 2-mercaptoethanol R and 3.1 g/L of - size: l = 0.035 m, 0 = 4.6 mm;
sodium dithíoníte R and incubate at 50 ºC for 3 h. Dilute to - stationary phase: resin for hydrophobic interaction
1 mL with water R. chromatography R (2.5 µm);
Reference solution (a). Dissolve the contents of a vial of - temperature: 35 ºC.
etanercept CRS in water R to obtain a concentration of about Mobile phase:
5 mg/mL. Carry out the release and labelling of sialic acid in
the same manner as for the test solution. Dilute to 1 mL with - mobile phase A: dissolve 28.4 g of anhydrous disodium
water R. hydrogen phosphate R and 475.9 g of ammonium sulfate R
in water for chromatography R and dilute to 1950 mL with
Reference solution (b ). Dissolve N-acetylneuraminic acid R the same solvent; adjust to pH 7.0 with phosphoric acid R
in water R to obtain a concentration of about 1 m2:/mL. Mix and dilute to 2000 mL with water for chromatography R;
40 µL of the solution, 40 µL of a 1 mg/mL solutio~ of bovine
albumin Rl and 120 µL of solution A. Use 50 µL of this - mobile phase B: dissolve 28.4 g of anhydrous disodium
solution to carry out the release and labelling of sialic acid in hydrogen phosphate R in water for chromatography R and
the same manner as for the test solution. Dilute with water R dilute to 1950 mL with the same solvent; adjust to pH 7.0
to obtain a concentration of 0.01 µg/µL. with phosphoric acid R and dilute to 2000 mL with water
f or chromatography R;
Standard solutions. Dilute reference solution (b) with water R
to obtain a concentration of 2 ng/µL. Further dilute this Time Mobile phase A Mobile phase B
solution to prepare a standard curve with concentrations (min) (per cent V/V) (per cent V/V)
in the range of 0.10-0.40 ng/µL (6 concentrations, typically o - 50 100 -7 o o -7 100
0.10 ng/µL, 0.15 ng/µL, 0.20 ng/µL, 0.25 ng/µL, 0.30 ng/µL,
0.40 ng/µL). Analyse the labelled sialic acid by liquid Flow rate: 1.0 mL/min.
chromatography (2.2.29). Detection: fluorimeter at 278 nm for excitation and 350 nm
Column: for emission.
- size: l = 0.25 m, 0 = 4.6 mm; Autosampler: set at 10 ºC.
- stationary phase: end-capped octadecylsilyl silica gel for Injection: 5 µL; perform 3 injections.
chromatography R (5 µm) with a pore size of 8 nm; Relative retention with reference to etanercept (retention
- temperature: 35 ºC. time= about 28.5 min): peak 1 = 0.96; peak 3 = 1.12.
Mobile phase: mix 8 mL of acetonitrile R, 500 mL of methanol R System suitability: reference solution:
and 1820 mL of water for chromatography R; mix thoroughly. - the chromatogram obtained is qualitatively similar to the
Flow rate: 1 mL/min. chromatogram supplied with etanercept CRS;
Detection: fluorimeter at 374 nm for excitation and 448 nm - peak-to-valley ratio: minimum 1.1, where HP = height above
for emission. the baseline of peak 1 and Hv = height above the baseline of
the lowest point of the curve separating this peak from the
Autosampler: set at 2-8 ºC. peak dueto etanercept; minimum 1.9, where HP = height
Injection: 20 µL. above the baseline of peak 3 and Hv = height above the
Retention time: sialic acid = about 10.5 min. baseline of the lowest point of the curve separating this
peak from the peak due to etanercept.
Calculate the sialic acid content in the preparation to be
examined using the standard curve and the content of sialic Results:
acid (N-acetylneuraminic acid) in the standard solutions. - no additional peaks are observed in the chromatogram
Report the molar ratio (number of moles of sialic acid per obtained with the test solution in comparison with the
mole of etanercept), using the molar mass of the monomer. chromatogram obtained with the reference solution.
Limits: Reference solution (e). Mix 1 volume ofreference solution (b)

- peak 1 : maximum 5 per cent; and 4 volumes of concentrated SDS-PAGE sample buffer R.
- peak 3: maximum 28 per cent; Reference solution (d). Mix 1 volume of reference solution (b)
- sum of ali peaks other than the principal peak: maximum and 19 volumes of concentrated SDS-PAGE sample buffer R.
30 per cent. Reference solution (e). A solution of molecular mass markers
suitable for calibrating SDS-polyacrylamide gels in the range
Impurities with molecular masses greater than that of of 5-200 kDa.
etanercept. Size-exclusion chromatography (2.2.30): use the
normalisation procedure. Sample treatment: heat at 90-105 ºC for 5 min and load onto
the gel within 15 min.
Solution A. Dissolve 8.8 g of sodium chloride R and 15.6 g
of sodium dihydrogen phosphate R in 900 mL of water for Application: 10 µL.
chromatography R and dilute to 1000 mL with the same Detection: by silver staining.
solvent. Identification of bands:
Solution B. Dissolve 8.75 g of sodium chloride R and 14.2 g - non-reducing conditions: the band corresponding to
of anhydrous disodium hydrogen phosphate R in 900 mL of etanercept of an apparent molecular mass of approximately
water for chromatography R and dilute to 1000 mL with the 150 kDa is present; related protein bands with apparent
same solvent. molecular masses of approximately 60 kDa, 100 J<_Da,
Test solution. Dilute the preparation to be examined with 120 kDa, 225 kDa, 250 kDa may also be present;
water R to obtain a concentration of 2.5 mg/mL. - reducing conditions: the band corresponding to etanercept
Reference solution. Dissolve the contents of a vial of of an apparent molecular mass of approximately 76 kDa
etanercept CRS in water R to obtain a concentration of is present; related protein bands with apparent molecular
2.5 mg/mL. masses of approximately 25 kDa, 35 kDa, 55 kDa, 200 kDa
Column: may also be present.
- size: l = 0.30 m, 0 = 8.0 mm; System suitability:
stationary phase: hydrophilic silica gel for chromatography R - the bands in the electropherogram obtained with reference
(5 µm) with a pore size of 30 nm and of a grade suitable for solution (a) are clearly visible;
fractionation of globular proteins in the relative molecular - the band in the electropherogram obtained with reference
mass range of 1O 000 to 1000 000. solutions (b), (c) and (d) is clearly visible;
Mobile phase: mix 220 mL of solution A and 780 mL of - all expected bands in the electropherogram obtained with
solution B, and adjust to pH 7.2 with solution A or solution B. reference solution (e) are visible and clearly separated.
Flow rate: 1.0 mL/min. Results:
Detection: spectrophotometer at 220 nm. - the electropherogram obtained with the test solution is
Autosampler: set at 2-8 ºC. similar to the electropherogram obtained with reference
solution (a);
Injection: 14 µL; perform at least 3 injections.
- the electropherogram obtained with the test solution shows
Relative retention with reference to etanercept monomer
no additional band that is more intense than that of the
(retention time= about 7.8 min): aggregates = 0.84; high
band in the electropherogram obtained with reference
molecular mass species = 0.89.
solution (d).
Any shoulder appearing on the descending part of the peak
due to etanercept monomer is included in its area. Microbiological contamination (2.6.12): maximum
1 CFU/mL.
- the chromatogram obtained is qualitatively similar to the ASSAY
chromatogram supplied with etanercept CRS; Protein (2.5.33, Method 1).
- resolution: minimum 1.7 between the peaks dueto the high Test solution. Dilute the preparation to be examined
molecular mass species and etanercept monomer; gravimetrically with a suitable buffer to obtain a concentration
- number of theoretical plates: minimum 3000 calculated for of 1.0 mg/mL. Prepare in triplicate.
the peak due to etanercept monomer. Reference solution. Dissolve the contents of a vial of
Limit: etanercept CRS in a suitable buffer to obtain a concentration
- sum of the peaks eluted befare the principal peak: maximum of 1.0 mg/mL.
8.0 per cent. Record the absorbance spectrum between 250 nm and
Impurities with molecular masses differing from that of 400 nm. Measure the value at the absorbance maximum of
etanercept. Polyacrylamide gel electrophoresis (2.2.31) under 280 nm after correction for any light scattering measured up
both reducing and non-reducing conditions. to 320 nm. Calculate the protein concentration of etanercept
taking into account the assigned content of etanercept CRS.
Gel dimensions: 1.0 mm thick.
Resolving gel: 8-16 per cent acrylamide. Potency. The potency of etanercept is determined by
comparison of dilutions of the test preparation with the
Sample buffer (non-reducing conditions): concentrated dilutions of etanercept BRP using a suitable cell-based assay
SDS-PAGE sample buffer R. based on the inhibitory action of etanercept on the biological
Sample buffer (reducing conditions): concentrated SDS-PAGE activity of TNF-a anda suitable readout for assessing this
sample buffer for reducing conditions R. inhibitory effect.
Test solution. Dilute the preparation to be examined with The following procedure has been found suitable.
water R to obtain a concentration of 0.2 mg/mL. Mix 1 volume Carry out an apoptosis-based assay based on the ability of
of this solution and 1 volume of sample buffer. etanercept to inhibit TNF-a induced apoptosis in histiocytic
Reference solution (a). Dissolve the contents of a vial of lymphoma cell line U937 (ATCC No. CRL-1593.2) via caspase
etanercept CRS in water R to obtain a concentration of activation. The U937 cells are incubated with varying dilutions
0.2 mg/mL. Mix 1 volume of the solution and 1 volume of of test and reference preparations of etanercept in the presence
sample buffer. of TNF-a. They are then incubated with Caspase-Glo 3/7
Reference solution (b). 0.01 mg/mL solution of bovine reagent, which results in caspase cleavage of a luminogenic
albumin R. substrate, subsequent release of a luciferase substrate and

generation of a luminescent signal. The luminescence Cell preparation. A cell density between 3.0 x 10 5 and 1.0 x 106
produced is proportional to the amount of caspase activity cells per millilitre is suitable, and cell viability is not less than
present. 95 per cent.
Assay medium. RPMI 1640 containing L-alanyl-L-glutamine, Plating test solution, reference solution, controls and cells.
6.0 g/L HEPES R (25 mM) and foetal bovine serum (7.5 per Transfer 60 µL from each cluster tube and add to the
cent V/V). corresponding wells. Mix the cell suspension thoroughly
Test solutions. Dilute the preparation to be examined with and add 60 µL to each well. Mix the contents of the plates
assay medium to obtain a concentration of about 72 ng/mL. on a shaker for 5 min. Incubate the plates without lids at
Use this solution to prepare 11 additional sample dilutions 36.0-38.0 ºC for 2-2.5 hin a humidified incubator using
(dilution steps of 1.2 or 1.4 have been found suitable). 5 ± 2 per cent C0 2•
Reference solutions. Dissolve the contents of a vial of Addition of Caspase-Glo 317 assay system. Reconstitute the
etanercept BRP in assay medium to obtain a concentration of Caspase-Glo 3/7 assay system according to the manufacturer's
about 72 ng/mL. Use this solution to prepare 11 additional instructions and add 100 µL to each well of the assay plates.
dilutions to generate the standard curve (dilution steps of 1.2 Shake the plates, covered with black lids, on a plate shaker
or 1.4 have been found suitable). for 10-15 min. Incubate at room temperature for 30-60 min.
TNF-a working solution. Dissolve the contents of a vial of Place the uncovered plates in a luminometer and read the
TNF-a according to the supplier's instructions. Further dilute luminescence for a minimum of 1 second per well.
with assay medium to obtain a suitable working concentration.
As the biological activity of TNF-a is likely to vary between Calculate the potency of the preparation to be examined using
different suppliers and also between different batches from the four-parameter logistic curve model (5.3).
the same supplier, this should be controlled by use of an System suitability:
appropriate standard (e.g. WHO International Standard for
TNF-a). - maximum value (TNF-a control) to mínimum value (cell
only) ratio: minimum 3.0.
Method.
Plate preparation. Add 600 µL of assay medium to the wells Result: the estimated potency is not less than 80 per cent and
designated for cells only (column l, rows A-D) on a cluster not more than 140 per cent relative to the reference solution.
tube rack. Add 300 µL of assay medium and 300 µL of TNF-a The confidence limits (P = 0.95) are not less than 80 per cent
working solution to the wells designated for the TNF-a and not more than 125 per cent of the estimated potency.
controls (column l, rows E-H). Add 300 µL of the test or
reference solutions and 300 µL of TNF-a working solution STORAGE
to the sample wells (columns 2-12, rows A-H). Mix on a
In an airtight container at - 20 ºC or below.
shaker for 5 min. Incubate at 36.0-38.0 ºC for 30-60 min in a
humidified incubator using 5 ± 2 per cent C0 2•
NOTE: when using deep-well or 96-well plates instead of cluster LABELLING
tubes, adapt the volumes of sample, TNF-a working solution The label states the content, in milligrams of protein per
and assay medium accordingly. millilitre.

F
Fipronil for veterinary use ..................................................... 5661 Follitropin ................................................................................ 5663
Folie acid hydrate .................................................................... 5662 Follitropin concentrated solution ......................................... 5669

EUROPEAN PHARMACOPOEIA 9.5 Fipronil for veterinary use
07/2018:2869 Mobile phase: tetrahydrofuran R, methanol R2, water for

chromatography R, acetonitrile Rl (O.S:30:30:40 V/V/V/V).
Detection: spectrophotometer at 21 O nm.
FIPRONIL FOR VETERINARY USE Injection: S µL of test solution (a) and reference solutions (a)
and (b).
Run time: twice the retention time of fipronil.
Fipronilum ad usum veterinarium
with fipronil for system suitability CRS and the chromatogram
obtained with reference solution (a) to identify the peaks due
to impurities A and B.
Relative retention with reference to fipronil (retention
time = about 6 min): impurity A = about 1.3;
impurity B = about 1.4.
C 12 H 4 Cl2 F6 N 4 0S Mr 437.l System suitability: reference solution (a):

[120068-37-3] - resolution: minimum 2.5 between the peaks due to
impurities A and B.
DEFINITION Calculation of percentage contents:
S-Amino-1- [2,6-dichloro-4-( trifluoromethyl)phenyl ]-4- [(RS)- - for each impurity, use the concentration of fipronil in
(trifluoromethyl)sulfinyl ]- lH-pyrazole-3-carbonitrile. reference solution (b).
Content: 9S.S per cent to 102.0 per cent (dried substance). Limits:
CHARACTERS - impurity B: maximum 3.S per cent;

Appearance: white or almost white or yellowish powder. - impurity A: maximum l .S per cent;
Solubility: practically insoluble in water, soluble in anhydrous - unspecified impurities: for each impurity, maximum
ethanol, practically insoluble in heptane. 0.20 per cent;
It shows polymorphism (5.9). - total: maximum 4.5 per cent;
IDENTIFICATION
Loss on drying (2.2.32): maximum O.S per cent, determined
Infrared absorption spectrophotometry (2.2.24). on 1.000 g by drying in an oven at lOS ºC.
Comparison: fipronil CRS. Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
If the spectra obtained in the solid state sho\A/ differences, 1.0 g in a platinum crucible.
dissolve the substance to be examined and the reference
substance separately in the minimum volume of methylene ASSAY
chloride R, evaporate to dryness and record new spectra using Liquid chromatography (2.2.29) as described in the test for
the residues. related substances with the following modification.
Injection: test solution (b) and reference solution (c).
TESTS
Related substances. Liquid chromatography (2.2.29). Carry Calculate the percentage content of C 12 H 4 Cl2 F6NPS taking
out the test protected from light. into account the assigned content of fipronil CRS.
Solvent mixture: methanol R, water R, acetonitrile R IMPURITIES

(30:30:40 V/V/V).
Specified impurities: A, B.
Test solution (a). Dissolve 3S.0 mg of the substance to be
examined in the solvent mixture and dilute to SO.O mL with
the solvent mixture.
Reference solution (a). Dissolve l.S mg of fipronil for system
suitability CRS (containing impurities A and B) in the solvent
mixture and dilute to 2.0 mL with the solvent mixture.
Reference solution (b ). Dilute 1.0 mL of test solution (a) to A. S-amino-1-[2,6-dichloro-4-( trifluoromethyl)phenyl]-4-
100.0 mL with the solvent mixture. Dilute 2.0 mL of this [(trifluoromethyl)sulfanyl]- lH-pyrazole-3-carbonitrile,
solution to 10.0 mL with the solvent mixture.
Reference solution (e). Dissolve 3S.O mg of fipronil CRS in
the solvent mixture and dilute to SO.O mL with the solvent
mixture. Dilute 3.0 mL of the solution to 20.0 mL with the
solvent mixture.
Column:
- size: l = O.lS m, 0 = 4.6 mm;
- stationary phase: end-capped octadecylsilyl silica gel for B. 5-amino-1-[2,6-dichloro-4-( trifluoromethyl)phenyl]-4-
chromatography R (1.8 µm). [(trifluoromethyl)sulfonyl]-lH-pyrazole-3-carbonitrile.
Folie acid hydrate EUROPEAN PHARMACOPOEIA 9.5
07/2018:0067 Reference solution (a). Dissolve SO.O mg of folie acid CRS in

2.5 mL of solution A and dilute to SO.O mL with the mobile
phase. Dilute 2.0 mL of this solution to 10.0 mL with the
mobile phase.
Reference solution (b ). Dissolve S mg of folie acid for system
FOLIC ACID HYDRATE suitability CRS (containing impurities C, E, G and H) in 1 mL
of solution A and dilute to 25.0 mL with the mobile phase.
Acidum folicum hydricum Reference solution (e). Dilute 1.0 mL of the test solution to
Reference solution (d). Dissolve 10.0 mg of folie acid
impurity A CRS in 1 mL of solution A and dilute to 100.0 mL
with the mobile phase. Dilute 1.0 mL of this solution to
100.0 mL with the mobile phase.
Reference solution (e). Dissolve 4.0 mg of folie acid
impurity D CRS in solution A and dilute to 100.0 mL with
C 19 H 19N 70 6 ,xH 20 Mr 441.4 (anhydrous substance) solution A. Dilute 1.0 mL of this solution to 100.0 mL with
Anhydrous folie acid: [S9-30-3] the mobile phase.
DEFINITION Reference solution (j). Dissolve 5 mg of folie acid for impurity I
identifieation CRS in 1 mL of solution A and dilute to 2S.O mL
(2S)-2-[ 4-[ [(2-Amino-4-oxo-1,4-dihydropteridin-6- with the mobile phase.
yl)methyl] amino] benzamido] pentanedioic acid hydrate.
Column:
- size: l = 0.2S m, 0 = 4.0 mm;
It contains a variable quantity of water.
- stationary phase: spherical octylsilyl silica gel for
CHARACTERS ehromatography R (S µm).
Appearance: yellowish or orange, crystalline powder. Mobile phase: mix 12 volumes of methanol R and 88 volumes
Solubility: practically insoluble in water and in most organic of a solution containing 11.16 g/L of potassium dihydrogen
solvents. It dissolves in dilute acids and in alkaline solutions. phosphate R and S.SO g/L of dipotassium hydrogen phosphate R.
IDENTIFICATION
First identification: A, B, D. Injection: 5 µL of the test solution and reference solutions (b ),
Second identifieation: A, C. (e), (d), (e) and (f).
A. Specific optical rotation (2.2.7): + 18 to+ 22 (anhydrous Run time: 3.3 times the retention time of folie acid.
substance). Identification of impurities: use the chromatogram obtained
Dissolve 0.25 g in a 4.2 g/L solution of sodium hydroxide R with reference solution (d) to identify the peak dueto
and dilute to 2S.O mL with the same solution. impurity A; use the chromatogram supplied with folie acid for
B. Infrared absorption spectrophotometry (2.2.24). system suitability CRS and the chromatogram obtained with
Comparison: folie acid CRS. reference solution (b) to identify the peaks due to impurities e,
E, G and H; use the chromatogram obtained with reference
C. Thin-layer chromatography (2.2.27).
solution (e) to identify the peak due to impurity D; use
Test solution. Dissolve SO mg of the substance to be the chromatogram supplied with folie acid for impurity I
examined in a mixture of 2 volumes of concentrated identification CRS and the chromatogram obtained with
ammonia R and 9 volumes of methanol R, and dilute to reference solution (f) to identify the peak due to impurity I.
100 mL with the same mixture of solvents.
Relative retention with reference to folie acid (retention
Reference solution. Dissolve SO mg of folie acid CRS in time = about 8.5 min): impurity A = about 0.5;
a mixture of 2 volumes of concentrated ammonia R and impurity C = about 0.9; impurity E = about 1.3;
9 volumes of methanol R, and dilute to 100 mL with the impurity D = about 1.5; impurity I = about 2.1 S;
same mixture of solvents. impurity G = about 2.4; impurity H = about 2.5.
Plate: TLC siliea gel plate R. System suitability: reference solution (b):
Mobile phase: concentrated ammonia R, propano[ R, ethanol - resolution: minimum 2.0 between the peaks due to folie
(96 per cent) R (20:20:60 V/V/V). acid and impurity E;
Application: 2 µL. - peak-to-valley ratio: minimum l.S, where HP = height
Development: over 3/4 of the plate. above the baseline of the peak due to impurity e and
Drying: in air. Hv = height above the baseline of the lowest point of the
Detection: examine in ultraviolet light at 365 nm. curve separating this peak from the peak dueto folie acid;
minimum 1.5, where HP = height above the baseline of the
Results: the principal spot in the chromatogram obtained peak due to impurity G and Hv = height above the baseline
with the test solution is similar in position, fluorescence of the lowest point of the curve separating this peak from
and size to the principal spot in the chromatogram obtained
the peak due to impurity H.
with the reference solution.
Caleulation of percentage eontents:
D. Water (see Tests).
- for impurity A, use the concentration of impurity A in
TESTS reference solution (d);
Related substances. Liquid chromatography (2.2.29). - for impurity D, use the concentration of impurity D in
Solution A: 28.6 g/L solution of sodium carbonate R. reference solution (e);
Test solution. Dissolve SO.O mg of the substance to be - for impurities other than A and D, use the concentration
examined in 2.5 mL of solution A and dilute to SO.O mL with of folie acid in reference solution (e).
the mobile phase. Dilute 2.0 mL of this solution to 10.0 mL Limits:
with the mobile phase. - impurity A: maximum 0.5 per cent;

EUROPEAN PHARMACOPOEIA 9.5 Follitropin
- impurity D: maximum 0.4 per cent;

- impurities C, E, G: for each impurity, maximum 0.3 per
cent;
- impurities H, J: for each impurity, maximum 0.15 per cent;
0.10 per cent;
Water (2.5.12): 5.0 per cent to 8.5 per cent, determined on E. (2S)-2-[4-[bis[(2-amino-4-oxo-l,4-dihydropteridin-6-yl)-
0.150 g. methyl] amino ]benzamido ]pentanedioic acid
Sulfated ash (2.4.14): maximum 0.2 per cent, determined on (6-pterinylfolic acid),
1.0 g.
ASSAY
related substances with the following modification.
Injection: test solution and reference solution (a).
F. 2-amino-7-(chloromethyl)pteridin-4(1H)-one,
STORAGE
Protected from light, under inert gas.
IMPURITIES
Specified impurities: A, C, D, E, G, H, J.
if present at a sufficient level, be detected by one or other of G. (2S)-[ 4-[(2-amino-7-methyl-4-oxo-1,4-dihydro-
the tests in the monograph. They are limited by the general pteridin-6-yl)amino ]benzamido ]pentanedioic acid,
for demonstration of compliance. See also 5.10. Control of
impurities in substances for pharmaceutical use): B, F.
H. (2S)-2-[ 4-[ (4S)-4-[ 4-[ [(2-amino-4-oxo-1,4-

A. (2S)-2-(4-aminobenzamido)pentanedioic acid dihydropteridin-6-yl)methyl] amino ]benzamido ]-4-
(N-( 4-aminobenzoyl)-L-glutamic acid), carboxybutanamido ]benzamido] pentanedioic acid,
l. unknown structure.
01/2015:2285
corrected 9.5
B. 2,5,6-triaminopyrimidin-4( lH)-one,
FOLLITROPIN
Follitropinum
u-subunit
APDVQDCPEC TLQENPFFSQ PGAPILQCMG CCFSRAYPTP 40
C. (2S)-2-[ 4-[[ (2-amino-4-oxo-1,4-dihydropteridin-7-yl)- LRSKKTMLVQ KNVTSESTCC VAKSYNRVTV MGGFKVENHT 80
methyl] amino ]benzamido] pentanedioic acid (isofolic ACHCSTCYYH KS 92
acid),
¡3-subunit
NSCELTNITI AIEKEECRFC ISINTTWCAG YCYTRDLVYK 40*
DPARPKIQKT CTFKELVYET VRVPGCAHHA DSLYTYPVAT 80*
QCHCGKCDSD STDCTVRGLG PSYCSFGEMK E 111*
glycosylation sites:
Asn-52, Asn-78, Asn-7*, Asn-24*
disulfide bridges:
7-31, 10-60, 28-82, 32-84, 59-87, 3*-51 *, 17*-66*, 20*-104*,
28*-82*, 32*-84*, 87*-94*
D. 4-[[ (2-amino-4-oxo-l,4-dihydropteridin-6-yl)-
methyl]amino ]benzoic acid (pteroic acid), Mr approx. 30 000 - 40 000
Follitropin EUROPEAN PHARMACOPOEIA 9.5
DEFINITION D. Peptide mapping (2.2.55).

Freeze-dried preparation of a heterodimeric glycoprotein SEPARA TION OF THE a- AND /3-SUBUNITS. Liquid
having the structure of human follicle-stimulating hormone chromatography (2.2.29).
(FSH). It consists of 2 subunits: a 92-amino-acid a-chain Test solution. Dissolve the substance to be examined
common to other glycoprotein hormones and a specific in mobile phase A to obtain a concentration of about
111-amino-acid ~-chain. 0.4 mg/mL.
Potency: 9000 JU to 17 000 IU per milligram of protein. Reference solution. Dilute follitropin for peptide mapping
and glycan analysis CRS with mobile phase A to obtain a
PRODUCTION concentration of about 0.4 mg/mL.
Follitropin is produced in mammalian cells by a method based Precolumn:
on recombinant DNA (rDNA) technology.
- size: l = 0.012 m, 0 = 4.6 mm;
Follitropin complies with the following requirements.
Host-cell-derived proteins. The limit is approved by the chromatography R (5 µm).
Column:
Host-cell- and vector-derived DNA. The limit is approved - size: l = 0.25 m, 0 = 4.6 mm;
CHARACTERS chromatography R (5 µm) with a pore size of 30 nm.
Appearance: white or almost white powder. Mobile phase:
- mobile phase A: dilute 1 mL of trifluoroacetic acid R to
IDENTIFICATION 1000 mL with water for chromatography R;
A. It complies with the requirements described under Assay. - mobile phase B: trifluoroacetic acid R, water for
B. Isoelectric focusing (2.2.54). chromatography R, aceto ni trile for chromatography R
Test solution. Dissolve the substance to be examined in (0.9:50:950 V/V/V);
water R to obtain a concentration of about 2 mg/mL, then Time Mobile phase A Mobile phase B
desalt and concentrate using a suitably validated procedure. (min) (per cent V/V) (per cent V!V)
Reconstitute the recovered material in water R to obtain a o- 2 100 o
concentration of 5 mg/mL.
2 -8 100 7 76 o7 24
Reference solution. Desalt and concentrate f ollitropin CRS
using a suitably validated procedure. Reconstitute the 8 - 17 76 24
recovered material in water R to obtain a concentration
17 - 36 76 7 70 24 7 30
of 5 mg/mL.
Focusing: 36 - 41 70 7 25 30 7 75
- pH gradient: a combination of ampholytes and electrode 41 - 46 25 75

buffers giving a functional separation in the isoelectric 46 - 47 25 7 100 75 7 o
point (pi) range of 3.5-5.5 is selected, as defined by
the system suitability criteria; where pre-cast gels are 47 - 57 100 o
employed, proprietary electrode solutions may be used
in conjunction; otherwise, suitable dilute mineral or Flow rate: 1.0 mL/min.
organic acids and bases are employed at pH levels Detection: spectrophotometer at 226 nm.
respectively lower and higher than the functional range
Injection: 800 µL.
of the ampholytes;
Retention time: ~-subunit = about 14 min; a-subunit = about
- catholyte: 20.0 g/L solution of glycine R; 30min.
- anolyte: solution containing 3.4 g/L of aspartic acid R Collect the fractions containing the a- and ~-subunits and
and 3.6 g/L of glutamic acid R, adjusted to pH 2.8-3.8; freeze-dry them.
- application: 1O µL. REDUCTION, MODIFICATION AND DESALTING OF
Detection: as described in 2.2.54. THE PURIFIED SUBUNITS
System suitability: Reduction and modification
- in the electropherogram obtained with the reference Solution A. Dilute 10 µL of tributylphosphine R to 2 mL
solution, the number of bands seen in the pi with propano[ R. Saturate with nitrogen.
region 3.5-5.5 corresponds to that shown in the Solution B. Dilute 20 µL of 4-vinylpyridine R to 200 µL with
electropherogram supplied with follitropin CRS; propano[ R. Saturate with nitrogen.
the distribution of bands in the pi region 3.5-5.5
is qualitatively similar to that shown in the Test solutions. Dissolve each of the a- and
electropherogram supplied with follitropin CRS. ~-subunit fractions obtained from the test
solution in the previous step in 300 µL of
Results: examine the electropherogram obtained with the guanidine-tris(hydroxymethyl)aminomethane-EDTA buffer
test solution; identify the bands observed by comparison solution pH 8.5 R and incubate at 37 ºC for 60 min in a
with the electropherogram obtained with the reference thermostatically controlled water-bath. Add 100 µL of
solution; the pattern of bands is qualitatively similar to that solution A, mix and saturate with nitrogen. Incubate at
seen with the reference solution. 37 ºC for 90 min. Add 10 µL of solution B, mix and saturate
C. Examine the chromatograms obtained in the test for with nitro gen. Incubate at 3 7 ºC for 45 min. Add 100 µL of
follitropin oligomers. a 1Oper cent V/V solution of trifluoroacetic acid R and mix.
Results: the principal peak in the chromatogram obtained Reference solutions. Prepare at the same time and in the
with the test solution is similar in retention time to the same manner as for the test solutions but using the a- and
principal peak in the chromatogram obtained with the ~-subunit fractions obtained from the reference solution
reference solution. in the previous step.

EUROPEAN PHARMACOPOEIA 9.5 Follitropin
Desalting - stationary phase: octadecylsilyl silica gel f or

Dilute the a- and ~-subunit test and reference solutions to chromatography R (5 µm) with a pore size of 30 nm.
840 µL with mobile phase A. Mobile phase:
Column: - mobile phase A: dilute 1 mL of trifluoroacetic acid R to
- size: l = 0.02 m, 0 = 4.6 mm; 1000 mL with water for chromatography R;
stationary phase: butylsilyl silica gel for chromatography R - mobile phase B: trifluoroacetic acid R, water f or
(5 µm).
chromatography R, acetonitrile Rl (1:300:700 V/V/V);
Mobile phase: Time Mobile phase A Mobile phase B
1000 mL with water for chromatography R; o- 7 100 o
- mobile phase B: trifluoroacetic acid R, water for 7 - 77 100 -7 30 o -7 70
chromatography R, acetonitrile for chromatography R 77 - 82 30 -7 o 70 -7 100
(1:300:700 V/V/V);
82 - 87 o 100
(min) (per cent V/V) (per cent V/V) 87 - 92 o -7 100 100 -7 o
0-7 100 o 92 - 107 100 o
7 - 27 100 -7 o o -7 100
27 - 27.01 o -7 100 100 -7 o Detection: spectrophotometer at 21 O nm.
27.01 - 32 100 o Injection: 400 µL.
System suitability:
Flow rate: 1.0 mL/min. a-subunit:
Detection: spectrophotometer at 226 nm. - the chromatogram obtained with the reference solution
Injection: 800 µL. is qualitatively similar to the chromatogram of follitropin
Por each solution the chromatogram shows a a-subunit digest supplied withfollitropin for peptide
principal peak due to the monovinylpyridine-modified mapping and glycan analysis CRS; both chromatograms
subunit and several minor peaks dueto the di- and show peaks dueto the L4, L6, L3, L5 and Ll-2/Ll
oligovinylpyridine-modified subunits. Only the fraction fragments;
containing the monovinylpyridine-modified subunit is - retention times obtained with the test and reference
used for digestion in the following step. solutions differ by not more than 5 per cent for
Retention time: a-subunit solution: monovinylpyridine- fragments L4, L6 and L3, not more than 3 per cent for
modified a-subunit = about 15 min; {3-subunit solution: fragment L5 and not more than 2 per cent for fragments
monovinylpyridine-modified ~-subunit = about 16 min. Ll-2/Ll;
Collect the fractions containing the monovinylpyridine- {3-subunit:
modified subunits and freeze-dry them. - the chromatogram obtained with the reference solution
SELECTIVE CLEA VAGE OF THE PEPTIDE BONDS is qualitatively similar to the chromatogram of follitropin
~-subunit digest supplied with follitropin for peptide
Solution C (8 M urea solution). Dissolve 480 g of urea R mapping and glycan analysis CRS; both chromatograms
in 600 mL of water for chromatography R and dilute show peaks dueto the L5, L7, L6, and Ll-4 fragments;
to 1000 mL with the same solvent. Add about 3-5 g of
mixed-bed resin and stir for about 1 h. Filter through a - retention times obtained with the test and reference
glass filter before use. solutions differ by not more than 5 per cent for fragment
L5, not more than 2 per cent for fragments L7 and L6
Solution D. Dissolve 15.8 g of ammonium hydrogen and not more than 1 per cent for fragments Ll-4.
carbonate R and 8.3 g of sodium edetate R in 800 mL of
water for chromatography R. Adjust to pH 7.8 (2.2.3) with Results: for each subunit, the profile of the chromatogram
an 80 g/L solution of sodium hydroxide R and dilute to obtained with the test solution is similar to that of the
1000 mL with water for chromatography R. chromatogram obtained with the corresponding reference
solution.
Test solutions. Dissolve each of the modified a- and
E. Glycan analysis (2.2.59). Carry out either method A or
~-subunits obtained from the test solutions in the previous
step in 42.5 µL of solution C and incubate at room method B.
temperature for 30 min. Add 42.5 µL of solution D and METHODA
mix. To 42.5 µL of these solutions add 35 µL of a solution PROTEIN DENATURATION
containing about 23 mU/µL of endoproteinase Lys-C and Test solution. Dissolve 500 µg of the substance to be
mix. Incubate at 37 ºC for 4 h, then add 35 µL of the same examined in 60 µL of 0.05 M phosphate buffer solution
endoproteinase Lys-C solution and mix. Incubate at 37 ºC pH 7.5 R. Add 6 µL of a 10 mg/mL solution of sodium
overnight, then dilute to 420 µL with mobile phase A. dodecyl sulfate R and 35 µL of a 1 per cent V/V solution of
Referen ce solutions. Prepare at the same time and in the 2-mercaptoethanol R. Mix using a vortex mixer, centrifuge
same manner as for the test solutions but using the fractions and incubate at 37 ºC for 15 min.
obtained from the reference solutions in the previous step. Reference solution. Freeze dry a sample of follitropin for
CHROMATOGRAPHIC SEPARATION. Liquid peptide mapping and glycan analysis CRS that contains
chromatography (2.2.29). 500 µg of follitropin. Dissolve in 60 µL of O. 05 M phosphate
Precolumn: buffer solution pH 7.5 R and continue as for the test
solution.
- size: l = 0.02 m, 0 = 4.6 mm;
SELECTIVE RELEASE OF THE GLYCANS
- stationary phase: octadecylsilyl silica gel f or
Test solution. To the test solution obtained in the previous
step add 0.75 µL of octylphenyl-polyethylene glycol and mix
Column: using a vortex mixer. Add 25 mU of peptide N-glycosidase
- size: l = 0.25 m, 0 = 4.6 mm; F R, mix using a vortex mixer and centrifuge. Incubate at
37 ºC for 24 h. Remove the protein fraction using a suitable,

validated procedure. The following method has been found (Ao x O)+ (A1 x 1) + (A2 x 2) + (A3 x 3) + (A4 x 4)
to be appropriate. Add 600 µL of anhydrous ethanol R,
previously cooled at - 20 ºC for 45 min. Mix using a vortex
mixer and centrifuge. Precipitate the proteins at - 20 ºC A0 peak area percentage due to the neutral
for 15 min, then centrifuge at 1O 600 g at 4 ºC for 5 min. form;
Transfer the supernatant to a separate tube and evaporate
the ethanol for 15 min. Add 1 mL of particle-free water R A1 peak area percentage dueto the
and resume evaporating until the remaining volume is mono-sialylated form;
about 500-800 µL, then freeze-dry.
A2 peak area percentage dueto the di-sialylated
Label the liberated glycans contained in the sample with forro;
2-aminobenzamide. The procedure employs a combination
of reagents optimised and validated for the efficient A3 peak area percentage due to the
labelling of glycans, and for the subsequent extraction and tri-sialylated form;
recovery of the labelled glycans from the reaction. Recover
the sample in 1.5 mL of particle-free water R. A4 peak area percentage due to the
tetra-sialylated form.
Reference solution. Prepare at the same time and in the
same manner as for the test solution but using the reference The Z number obtained for the reference solution is in
solution obtained in the previous step. the range 177-233.
CHROMATOGRAPHIC SEPARA TION. Liquid Examine the chromatogram obtained with the test solution
chromatography (2.2.29). and calculate the Z number as described above.
Result: Z = 177-233.
Column: METHOD B
- size: l = 0.075 m, 0 = 7.5 mm; PROTEIN DENATURA TION
- stationary phase: weak anion-exchange resin R (10 µm); Solution A. To 1.952 g of 2-[N-morpholino]ethanesulfonic
acid R and 57.32 g of guanidine hydrochloride R, add 1 mL
- temperature: 30 ºC. of a 15.4 g/L solution of dithiothreitol R, 10 mL of an
18.61 g/L solution of sodium edetate R and 20 mL of water
Mobile phase: for chromatography R. Maintain in a water-bath at about
37 ºC for 1 min to dissolve the components. Adjust to
- mobile phase A: acetonitrile R; pH 8.1 (2.2.3) with an 80 g/L solution of sodium hydroxide R
and dilute to 100.0 mL with water for chromatography R.
- mobile phase B: 0.5 M ammonium acetate buffer solution Mix.
pH 4.5 R; filter through a membrane filter (nominal
pore size 0.22 µm); Solution B. Dissolve 37 mg of iodoacetamide R in 1 mL of
water for chromatography R and mix. Protect from light.
- mobile phase C: particle-free water R; Solution C. Dissolve 26. 7 g of disodium hydrogen phosphate
dihydrate R and 11.2 g of sodium edetate R in 3000 mL of
Time Mobile phase A Mobile phase B Mobile phase C water for chromatography R and mix. Adjust to pH 7.5
(min) (per cent V/V) (per cent V/V) (per cent V/V) (2.2.3) with a 40 g/L solution of sodium hydroxide R.
o- 5 20 o 80 Test solution. Dissolve 1 mg of the substance to be examined
5 - 21 20 074 80 7 76 in 0.2 mL of solution A and incubate in a water-bath at
37 ± 1 ºC for 2 h. Add 20 µL of freshly prepared solution B,
21 - 61 20 4 7 25 76 7 55 mix and incubate at 37 ± 1 ºC for a further 2 h, protected
61 - 62 20 25 7 50 55 7 30 from light. Add 10 µL of 2-mercaptoethanol R and mix.
Dialyse against 1000 mL of solution C. Add 200 µL of
62 - 71 20 50 30 solution C and mix. Determine the protein content of the
71 - 72 20 50 7 o 30 7 80
solution.
Reference solution (a). To a volume of follitropin for peptide
72 - 117 20 o 80
mapping and glycan analysis CRS that contains 1 mg
of follitropin, add 0.2 mL of solution A. Incubate in a
Flow rate: 0.4 mL/min. water-bath at 37 ± 1 ºC for 2 h. Continue as for the test
solution. Determine the protein content of the solution.
Detection: tluorimeter at 330 nm for excitation and at
420 nm for emission. Reference solution (b ). Prepare at the same time and in
the same manner as for the test solution but using fetuin
Injection: 50 µL. instead of the substance to be examined. Determine the
protein content of the solution.
System suitability: reference solution: SELECTIVE RELEASE OF THE GLYCANS
- the chromatogram obtained is qualitatively similar to Test solution. Dilute the test solution obtained in the
the chromatogram supplied with follitropin for peptide previous step with solution C to obtain a concentration of
mapping and glycan analysis CRS; 1.1 g/L. Add 1 U of peptide N-glycosidase F R to 500 µg of
the solution, mix and incubate at 37 ± 1 ºC for 24 h. Place
- by comparison with the chromatogram supplied with the solution in ice. Precipitate the protein and salts with
follitropin for peptide mapping and glycan analysis CRS, 3 volumes of ice-cold anhydrous ethanol R and allow to
identify the peaks dueto neutral, mono-, di-, tri- and stand in ice for 10 min. Centrifuge at 16 000 g for about
tetra-sialylated forros; determine the area of each peak 5 min and transfer the supernatant to a separate tube. Add
and express it as a percentage of the total; calculate the 3 µL of a 1 µg/µL solution of maltotriose R, then freeze-dry.
Z number using the following expression: Dissolve in 100 µL of water for chromatography R.

EUROPEAN PHARMACOPOEIA 9.S Follitropin
Reference solution (a). Prepare in the same manner as for

the test solution but using the reference solution obtained (Ao x O) + (A1 x 1) + (A2 x 2) + (A3 x 3) + (A4 x 4)
with follitropin f or peptide mapping and glycan analysis CRS
in the previous step.
A0 peak area percentage due to the neutral
Referen ce solution (b). Prepare in the same manner as for form;
the test solution but using the reference solution obtained
with fetuin in the previous step. A1 peak area percentage due to the
CHROMATOGRAPHIC SEPARA TION. Liquid mono-sialylated form;
A2 peak area percentage dueto the di-sialylated
Precolumn: form;
- size: l =O.OS m, 0 = 4.0 mm; peak area percentage due to the
- stationary phase: strongly basic anion-exchange resin for tri-sialylated form;
chromatography R;
peak area percentage due to the
Column: tetra-sialylated form.
- size: l = 0.2S m, 0 = 4.0 mm; The Z number obtained for reference solution (b) is in
- stationary phase: strongly basic anion-exchange resin for the range 290-32S.
chromatography R. Examine the chromatogram obtained with the test solution
and calculate the Z number as described above.
Mobile phase:
Result: Z = 178-274.
- mobile phase A: 20 g/L solution of sodium hydroxide R;
maintain under helium; TESTS
- mobile phase B: water for chromatography R; maintain Follitropin oligomers. Size-exclusion chromatography
under helium; (2.2.30). Use the normalisation procedure.
- mobile phase C: dissolve 41 g of anhydrous sodium Solution A. Dissolve 118 mg of sodium dihydrogen phosphate R,
l.6S g of disodium hydrogen phosphate dihydrate R and 30.0 g
acetate R in 800 mL of water for chromatography R,
dilute to 1000 mL with the same solvent, then mix; filter of sucrose R in 40 mL of water for chromatography R and
through a membrane filter (nominal pore size 0.4S µm); dilute to 100.0 mL with the same solvent.
maintain under helium. Solution B. Dissolve 2.0 mg of bovine albumin R in 30 mL of
solution A.
Time Mobile phase A Mobile phase B Mobile phase C Test solution. Dissolve the substance to be examined in
(min) (per cent V/V) (per cent V/V) (per cent V/V) solution A to obtain a concentration of 0.2S mg/mL.
o - 0.2 20 80 o Reference solution. Dilute follitropin CRS with solution A to
0.2 - 94.0 20 80 -7 34 u -7 46 obtain a concentration of 0.5 mg/mL and mix equal volumes
of this solution and solution B to obtain a concentration of
94.0 - 97.0 20 34 46
0.2S mg/mL.
97.0 - 97.l 20 34 -7 80 46 -7 o Column:
97.1 - 115.0 20 80 o - size: l = 0.3 m, 0 = 7.8 mm;
- stationary phase: hydrophilic silica gel for chromatography R,
Flow rate: l.O mL/min. of a grade suitable for fractionation of globular proteins
in the relative molecular mass range of 10 000 to SOO 000
Detection: pulsed amperometric detector or equivalent with
(S µm).
a gold indicator electrode, a silver-silver chloride reference
electrode, and a stainless steel auxiliary electrode which Mobile phase: dissolve 28.4 g of anhydrous sodium sulfate R in
is the cell body, held at respectively + O.OS V detection, 2000 mL of 0.1 M phosphate buffer solution pH 6.7 R and filter
+ O. 7S V oxidation and - 0.80 V reduction potentials, with through a membrane filter (nominal pore size 0.4S µm).
pulse durations according to the instrument used. Flow rate: O.S mL/min.
Injection: 4S µL. Detection: spectrophotometer at 21 S nm.
System suitability: Injection: 100 µL.
Retention time: follitropin = 14-16 min.
- the chromatogram obtained with reference solution (b)
is qualitatively similar to the chromatogram for fetuin System suitability: reference solution:
supplied with follitropin for peptide mapping and glycan - resolution: mínimum 1.2 between the peaks due to bovine
analysis CRS; albumin and follitropin;
the chromatograms obtained with the test solution - no peak is detected between S min and 16 min in blank
and reference solution (a) are qualitatively similar to injections.
the chromatogram supplied with follitropin for peptide Limit:
mapping and glycan analysis CRS; - sum of the peaks with a retention time less than that of the
- by comparison with the chromatogram supplied with principal peak: maximum 0.5 per cent.
follitropin for peptide mapping and glycan analysis CRS, Free subunits. Polyacrylamide gel electrophoresis (2.2.31)
identify the peaks dueto neutral, mono-, di-, tri- and under non-reducing conditions.
tetra-sialylated forms in the chromatogram obtained Gel dimensions: l.S mm thick.
with reference solution (b); determine the area of each
peak and express it as a percentage of the total; calculate Resolving gel: 12 per cent acrylamide.
the Z number using the following expression: Sample buffer. Concentrated SDS-PAGE sample buffer R.
Test solution. Dissolve the substance to be examined in Time Mobile phase A Mobile phase B Mobile phase C
water R to obtain a concentration of 2 µg/µL. To 55 µL of the (min) (per cent V/V) (per cent V/V) (per cent V/V)
solution add 55 µL of the sample buffer. Allow to stand for 4 h o - 8.4 50 25 7 39 25 7 11
at room temperature.
8.4 - 8.5 50 39 7 45 11 7 5
Reference solution (a). Concentrate follitropin CRS using a
suitably validated procedure to obtain a concentration of 8.5 - 15 50 45 5
2 µg/µL. To 25 µL of the solution add 25 µL of the sample 15 - 15.l 50 45 7 25 5 7 25
buffer. To 40 µL of this solution add 180 µL of the sample
buffer and 180 µL of water R. Allow to stand for 4 h at room 15.l - 25 50 25 25
temperature, then boil for 5 min.
Reference solution (b ). A solution of molecular mass markers
suitable for calibrating SDS-polyacrylamide gels in the range Detection: spectrophotometer at 210 nm.
of 14.4-94 kDa. Injection: 25 µL.
Application: System suitability: reference solution (b):
- the peaks due to the oxidised follitropin a- and
Well Solution(s) Volume (µL) ~-subunits are separated from the peaks due to the
Reference solution (a) 40 non-oxidised follitropin subunits and from the peak due
to 2,4-dichlorobenzoic acid;
2 Reference solution (a) 30
- the chromatogram obtained is similar to the chromatogram
3 Reference solution (a) 20 supplied with follitropin CRS.
4 Reference solution (a) 15 Calculate the percentage of oxidation of the follitropin
subunits using the following expression:
(A2 + A4) X 100
Ai + A2 + A3 + A4
7 Test solution 50
8 Test solution + reference solution (a) 50 + 25 area of the peak due to the follitropin a-subunit;
9 Reference solution (b) 10 area of the peaks due to the oxidised follitropin
a-subunit;
Detection: by Coomassie staining. area of the peak due to the follitropin ~-subunit;
System suitability: area of the peak duc to the oxidised follitropin
- reference solution (b): the validation criteria are met ~-subunit.
(2.2.31); Limit:
- test solution + reference solution (a): the bands - total oxidised f orms: maximum 6 per cent.
corresponding to the follitropin heterodimer and subunits
are clearly separated; Bacteria} endotoxins (2.6.14): less than 0.1 IU per
International Unit of follitropin activity, if intended for use in
- reference solution (a): no bands corresponding to the the manufacture of parenteral preparations without a further
follitropin heterodimer are seen. appropriate procedure for the removal of bacteria! endotoxins.
Limit:
ASSAY
- free subunits: maximum 3 per cent.
Protein. Size-exclusion chromatography (2.2.30).
Oxidised follitropin. Liquid chromatography (2.2.29).
Solution A. Dissolve 100 mg of poloxamer 188 R in 900 mL of
Solution A. Dissolve about 3.3 mg of 2,4-dichlorobenzoic water for chromatography R and dilute to 1000 mL with the
acid R in 10.0 mL of ethanol (96 per cent) R. same solvent.
Test solution. Dissolve the substance to be examined in water Test solution. Dissolve the substance to be examined in
for chromatography R to obtain a concentration of 300 µg/mL. solution A to obtain a concentration of about 0.03 mg/mL.
Reference solution (a). Dilute follitropin CRS with water for Reference solution. Dilute follitropin CRS with solution A to
chromatography R to obtain a concentration of 300 µg/mL. obtain a concentration of about 0.03 mg/mL.
Reference solution (b ). Dilute 0.1 mL of strong hydrogen Column:
peroxide solution R to 30 mL with water for chromatography R. - size: l = 0.3 m, 0 = 7.8 mm;
Dilute follitropin CRS with this solution to obtain a - stationary phase: hydrophilic silica gel for chromatography R,
concentration of 300 µg/mL. Incubate for 30-45 min. Add of a grade suitable for fractionation of globular proteins
solution A to obtain a concentration in 2,4-dichlorobenzoic in the relative molecular mass range of 10 000 to 500 000
acid of about 17 µg/mL in the total volume and inject (5 µm).
immediately.
Mobile phase: mix 6. 74 mL of phosphoric acid R, 14.2 g
Column: of anhydrous sodium sulfate R and 900 mL of water for
- size: l = 0.25 m, 0 = 4.6 mm; chromatography R, adjust to pH 6.7 (2.2.3) with a 0.5 g/mL
solution of sodium hydroxide R and dilute to 1000 mL with
chromatography R (5 µm); water for chromatography R; :filter through a membrane filter
(nominal pore size 0.45 µm).
- tempera tu re: 30 ºC. Flow rate: 1 mL/min.
Mobile phase: Detection: spectrophotometer at 214 nm.
- mobile phase A: 0.2 M phosphate buffer solution pH 2.5 R; Injection: 100 µL.
- mobile phase B: water for chromatography R, acetonitrile Rl System suitability: reference solution:
(40:60 V/V); - number of theoretical plates: minimum 1300, calculated for
- mobile phase C: water for chromatography R; the peak due to follitropin.

EUROPEAN PHARMACOPOEIA 9.5 Follitropin concentrated solution
Calculate the content of follitropin taking into account the 01/2015:2286

assigned content of follitropin CRS. corrected 9.5
Potency
The follicle-stimulating activity of follitropin is estimated by
comparing under given conditions its effect in enlarging the
ovaries of immature rats treated with chorionic gonadotrophin
with the same effect of the International Standard preparation FOLLITROPIN CONCENTRATED
of human recombinant follicle-stimulating hormone or of SOLUTION
a reference preparation calibrated in International Units.
The International Unit of FSH is the activity contained in
stated amounts of the International Standard of human Follitropini solutio concentrata
recombinant follicle-stimulating hormone. The equivalence in
International Units of the International Standard is stated by a-subunit
the World Health Organization. APDVQDCPEC TLQENPFFSQ PGAPILQCMG CCFSRAYPTP 40
LRSKKTMLVQ KNVTSESTCC VAKSYNRVTV MGGFKVENHT 80
Use immature female rats of the same strain, 19-28 days old, ACHCSTCYYH KS 92
differing in age by not more than 3 days and having masses J3-subunit
such that the difference between the heaviest and the lightest NSCELTNITI AIEKEECRFC ISINTTWCAG YCYTRDLVYK 40*
rat is not more than 10 g. Assign the rats at random to 6 equal DPARPKIQKT CTFKELVYET VRVPGCAHHA DSLYTYPVAT 80*
groups of at least 5 rats. If sets of 6 litter mates are available, QCHCGKCDSD STDCTVRGLG PSYCSFGEMK E 111*
assign 1 litter mate from each set to each group and mark glycosylation sites:
according to litter. Asn-52, Asn-78, Asn-7*, Asn-24*
disulfide bridges:
Choose 3 doses of the reference preparation and 3 doses of 7-31, 10-60, 28-82, 32-84, 59-87, 3*-51 *, l 7*-66*, 20*-104*,
the preparation to be examined such that the smallest <lose 28*-82*, 32*-84*, 87*-94*
produces a positive response in sorne of the rats and the
Mr approx. 30 000 - 40 000
largest <lose <loes not produce a maximal response in all of
the rats. Use doses in geometric progression and as an initial
approximation total doses of 1.5 IU, 3.0 IU and 6.0 IU may be DEFINITION
tried, although the <lose will depend on the sensitivity of the Solution of a heterodimeric glycoprotein having the structure
rats used, which may vary widely. of human follicle-stimulating hormone (FSH). It consists
of 2 subunits: a 92-amino-acid a-chain common to other
Dissolve separately the total quantities of the preparation to glycoprotein hormones and a specific 111-amino-acid ~-chain.
be examined and of the reference preparation corresponding
to the daily doses to be used in sufficient phosphate-albumin Content: 0.4 mg to 0.8 mg of protein per millilitre.
buffered saline pH 7.2 R such that the daily <lose is administered Potency: 9000 IU to 17 000 IU per milligram of protein.
in a volume of about 0.5 mL. The buffer solution shall
contain in the daily <lose not less than 14 IU of chorionic
PRODUCTION
gonadotrophin to ensure complete luteinisation. i\.dd a
suitable antimicrobial preservative such as 4 g/L of phenol or Follitropin is produced in mammalian cells by a method based
0.02 g/L of thiomersal. Store the solutions at 5 ± 3 ºC. on recombinant DNA (rDNA) technology.
Inject subcutaneously into each rat the daily <lose allocated to Follitropin complies with the following requirements.
its group. Repeat the injection of each <lose 24 h and 48 h after Host-cell-derived proteins. The limit is approved by the
the 1st injection. About 24 h after the last injection, euthanise competent authority.
the rats and remove the ovaries from each rat. Remove any Host-cell- and vector-derived DNA. The limit is approved
extraneous fluid and tissue from the ovaries and weigh the by the competent authority.
2 combined ovaries of each rat immediately. Calculate the
results by the usual statistical methods (for example, 5.3),
using the mass of the 2 combined ovaries as the response. (The CHARACTERS
precision of the assay may be improved by a suitable correction Appearance: clear or slightly turbid, colourless liquid.
of the organ mass with reference to the mass of the rat from
which it was taken; an analysis of covariance may be used.) IDENTIFICATION
The estimated potency is not less than 80 per cent and not A. It complies with the requirements described under Assay.
more than 125 per cent of the stated potency. The confidence B. Isoelectric focusing (2.2.54).
limits (P = 0.95) of the estimated potency are not less than
64 per cent and not more than 156 per cent of the stated Test solution. Desalt and concentrate the preparation
potency. to be examined using a suitably validated procedure.
Reconstitute the recovered material in water R to obtain a
concentration of 5 mg/mL.
STORAGE Reference solution. Desalt and concentrate follitropin CRS
In an airtight container, at a temperature not exceeding using a suitably validated procedure. Reconstitute the
- 20 ºC. recovered material in water R to obtain a concentration
of 5 mg/mL.
Focusing:
LABELLING
- pH gradient: a combination of ampholytes and electrode
The label states: buffers giving a functional separation in the isoelectric
- the potency in International Units per milligram of protein; point (pl) range of 3.5-5.5 is selected, as defined by
the system suitability criteria; where pre-cast gels are
- where applicable, that the substance is suitable for use in employed, proprietary electrode solutions may be used
the manufacture of parenteral preparations. in conjunction; otherwise, suitable dilute mineral or
Follitropin concentrated solution EUROPEAN PHARMACOPOEIA 9.5
organic acids and bases are employed at pH levels Injection: 800 µL.
respectively lower and higher than the functional range Retention time: P-subunit = about 14 min; a-subunit = about
of the ampholytes; 30min.
- catholyte: 20.0 g/L solution of glycine R; Collect the fractions containing the a- and P-subunits and
- anolyte: solution containing 3.4 g/L of aspartic acid R freeze-dry them.
and 3.6 g/L of glutamic acid R, adjusted to pH 2.8-3.8; REDUCTION, MODIFICATION AND DESALTING OF
- application: 1O µL. THE PURIFIED SUBUNITS
Detection: as described in 2.2.54. Reduction and modification
System suitability: Solution A. Dilute 1O µL of tributylphosphine R to 2 mL
with propano[ R. Saturate with nitrogen.
- in the electropherogram obtained with the reference
solution, the number of bands seen in the pl Solution B. Dilute 20 µL of 4-vinylpyridine R to 200 µL with
region 3.5-5.5 corresponds to that shown in the propano[ R. Saturate with nitrogen.
electropherogram supplied with follitropin CRS; Test solutions. Dissolve each of the a- and
the distribution of bands in the pl region 3.5-5.5 P-subunit fractions obtained from the test
is qualitatively similar to that shown in the solution in the previous step in 300 µL of
electropherogram supplied with follitropin CRS. guanidine-tris(hydroxymethyl)aminomethane-EDTA buffer
Results: examine the electropherogram obtained with the solution pH 8.5 R and incubate at 37 ºC for 60 minina
test solution; identify the bands observed by comparison thermostatically controlled water-bath. Add 100 µL of
with the electropherogram obtained with the reference solution A, mix and saturate with nitrogen. Incubate at
solution; the pattern of bands is qualitatively similar to that 37 ºC for 90 min. Add 10 µL of solution B, mix and saturate
seen with the reference solution. with nitro gen. Incubate at 37 ºC for 45 min. Add 100 µL of
a 1O per cent V/V solution of trifluoroacetic acid R and mix.
C. Examine the chromatograms obtained in the test for
follitropin oligomers. Reference solutions. Prepare at the same time and in the
same manner as for the test solutions but using the a- and
Results: the principal peak in the chromatogram obtained P-subunit fractions obtained from the reference solution
with the test solution is similar in retention time to the in the previous step.
principal peak in the chromatogram obtained with the
reference solution. Desalting
D. Peptide mapping (2.2.55). Dilute the a- and p-subunit test and reference solutions to
840 µL with mobile phase A.
SEPARATION OF THE a- AND /3-SUBUNITS. Liquid
chromatography (2.2.29). Column:
• 1 A I"\...... /"X Al F
- szze: t = u.u.L. m, \U= '±.o mm;
Test solution. Dilute the preparation to be examined
with mobile phase A to obtain a concentration of about - stationary phase: butylsilyl silica gel for chromatography R
0.4 mg/mL. (5 µm).
Reference solution. Dilute follitropin for peptide mapping Mobile phase:
and glycan analysis CRS with mobile phase A to obtain a - mobile phase A: dilute 1 mL of trifluoroacetic acid R to
concentration of about 0.4 mg/mL 1000 mL with water for chromatography R;
Precolumn: - mobile phase B: trifluoroacetic acid R, water f or
- size: l = 0.012 m, 0 = 4.6 mm; chromatography R, acetonitrile for chromatography R
(1:300:700 V/V/V);
chromatography R (5 µm). Time Mobile phase A Mobile phase B
Column:
0-7 100 o
- size: l = 0.25 m, 0 = 4.6 mm;
- stationary phase: end-capped butylsilyl silica gel f or 7 - 27 100 7 o o7 100
chromatography R (5 µm) with a pore size of 30 nm. 27 - 27.01 o -7 100 100 -7 o

Mobile phase: 27.01 - 32 100 o
1000 mL with water for chromatography R; Flow rate: 1.0 mL/min.
- mobile phase B: trifluoroacetic acid R, water f or Detection: spectrophotometer at 226 nm.
chromatography R, acetonitrile for chromatography R Injection: 800 µL.
(0.9:50:950 V/V/V); For each solution the chromatogram shows a
Time Mobile phase A Mobile phase B principal peak dueto the monovinylpyridine-modified
(min) (per cent V/V) (per cent V/V) subunit and several minor peaks dueto the di- and
0-2 100 o oligovinylpyridine-modified subunits. Only the fraction
containing the monovinylpyridine-modified subunit is
2-8 100 -7 76 o -7 24 used for digestion in the following step.
8 - 17 76 24 Retention time: a-subunit solution: monovinylpyridine-
modified a-subunit = about 15 min; f3-subunit solution:
17 - 36 76 -7 70 24 7 30
monovinylpyridine-modified P-subunit = about 16 min.
36 - 41 70 -7 25 30 7 75 Collect the fractions containing the monovinylpyridine-
41 - 46 25 75 modified subunits and freeze-dry them.
SELECTIVE CLEA VAGE OF THE PEPTIDE BONDS
46 - 47 25 -7 100 75 7 o
Solution C (8 M urea solution). Dissolve 480 g of urea R
47 - 57 100 o in 600 mL of water for chromatography R and dilute
to 1000 mL with the same solvent. Add about 3-5 g of
Flow rate: 1.0 mL/min. mixed-bed resin and stir for about 1 h. Filter through a
Detection: spectrophotometer at 226 nm. glass filter before use.

Solution D. Dissolve 15.8 g of ammonium hydrogen {3-subunit:

carbonate R and 8.3 g of sodium edetate R in 800 m1 of - the chromatogram obtained with the reference solutions
water for chromatography R. Adjust to pH 7.8 (2.2.3) with is qualitatively similar to the chromatogram of follitropin
an 80 g/1 solution of sodium hydroxide R and dilute to ~-subunit digest supplied with f ollitropin f or peptide
1000 m1 with water for chromatography R. mapping and glycan analysis CRS; both chromatograms
Test solutions. Dissolve each of the modified a- and show peaks dueto the L5, 17, 16, and Ll-4 fragments;
~-subunits obtained from the test solutions in the previous - retention times obtained with the test and reference
step in 42.5 µL of solution C and incubate at room solutions differ by not more than 5 per cent for fragment
temperature for 30 min. Add 42.5 µL of solution D and 15, not more than 2 per cent for fragments 17 and 16
mix. To 42.5 µ1 of these solutions add 35 µ1 of a solution and not more than 1 per cent for fragments Ll-4.
containing about 23 mU/µ1 of endoproteinase Lys-C and
Results: for each subunit, the profile of the chromatogram
mix. Incubate at 37 ºC for 4 h, then add 35 µ1 of the same
obtained with the test solution is similar to that of the
endoproteinase 1ys-C solution and mix. Incubate at 37 ºC
chromatogram obtained with the corresponding reference
overnight, then dilute to 420 µ1 with mobile phase A.
solution.
Reference solutions. Prepare at the same time and in the E. Glycan analysis (2.2.59). Carry out either method A or
same manner as for the test solutions but using the fractions method B.
obtained from the reference solutions in the previous step.
METHODA
CHROMATOGRAPHIC SEPARATION. Liquid PROTEIN DENATURATION
Test solution. Freeze-dry a sample of the preparation to
Precolumn: be examined that contains 500 µg of follitropin. Dissolve
in 60 µ1 of 0.05 M phosphate buffer solution pH 7.5 R. Add
- size: l = 0.02 m, 0 = 4.6 mm; 6 µL of a 10 mg/mL solution of sodium dodecyl sulfate R and
- stationary phase: octadecylsilyl silica gel for 35 µ1 of a 1 per cent V/V solution of 2-mercaptoethanol R.
chromatography R (5 µm). Mix using a vortex mixer, centrifuge and incubate at 37 ºC
for 15 min.
Column: Reference solution. Prepare at the same time and in the
- size: l = 0.25 m, 0 = 4.6 mm; same manner as for the test solution but usingfollitropin
for peptide mapping and glycan analysis CRS instead of the
- stationary phase: octadecylsilyl silica gel f or freeze-dried preparation to be examined.
chromatography R (5 µm) with a pore size of 30 nm. SELECTIVE RELEASE OF THE GLYCANS
Mobile phase: Test solution. To the test solution obtained in the previous
step add 0.75 µ1 of octylphenyl-polyethylene glycol and mix
- mobile phase A: dilute 1 m1 of trifluoroacetic acid R to using a vortex mixer. Add 25 mU of peptide N-glycosidase
1000 m1 with water for chromatography R; F R, mix using a vortex mixer and centrifuge. Incubate at
- mobile phase B: trifluoroacetic acid R, water for 37 ºC for 24 h. Remove the protein fraction using a suitable,
chromdtography R: acetonitrile Rl ( 1:300:700 V/V/V); vaiidated procedure. The following method has been found
to be appropriate. Add 600 µL of anhydrous ethanol R,
Time Mobile phase A Mobile phase B previously cooled at - 20 ºC for 45 min. Mix using a vortex
(min) (per cent V/V) (per cent V/V) mixer and centrifuge. Precipitate the proteins at - 20 ºC
0-7 100 o for 15 min, then centrifuge at 10 600 g at 4 ºC for 5 min.
Transfer the supernatant to a separate tube and evaporate
7 - 77 100 -7 30 o -7 70 the ethanol for 15 min. Add 1 ml of particle-free water R
77 - 82 30 -7 o 70 -7 100 and resume evaporating until the remaining volume is
about 500-800 µ1, then freeze-dry.
82 - 87 o 100
1abel the liberated glycans contained in the sample with
87 - 92 o -7 100 100 -7 o 2-aminobenzamide. The procedure employs a combination
of reagents optimised and validated for the efficient
92 - 107 100 o
labelling of glycans, and for the subsequent extraction and
recovery of the labelled glycans from the reaction. Recover
Flow rate: 1.0 mL/min. the sample in 1.5 mL of particle-free water R.
Detection: spectrophotometer at 210 nm. Reference solution. Prepare at the same time and in the
same manner as for the test solution but using the reference
Injection: 400 µL. solution obtained in the previous step.
System suitability: CHROMATOGRAPHIC SEPARA TION. Liquid
a-subunit:
Column:
- the chromatogram obtained with the reference solution size: l = 0.075 m, 0 = 7.5 mm;
is qualitatively similar to the chromatogram of follitropin
a-subunit digest supplied with follitropin for peptide - stationary phase: weak anion-exchange resin R (1 O µm);
mapping and glycan analysis CRS; both chromatograms - temperature: 30 ºC.
show peaks dueto the 14, 16, L3, L5 and Ll-2/Ll
Mobile phase:
fragments;
- mobile phase A: acetonitrile R;
- retention times obtained with the test and reference
solutions differ by not more than 5 per cent for - mobile phase B: 0.5 M ammonium acetate buffer solution
fragments 14, 16 and 13, not more than 3 per cent for pH 4.5 R; filter through a membrane filter (nominal
fragment L5 and not more than 2 per cent for fragments pore size 0.22 µm);
Ll-2/Ll; - mobile phase C: particle-free water R;
Time Mobile phase A Mobile phase B Mobile phase C 2 h. Add 20 µL of freshly prepared solution B, mix and
(min) (~er cent V/V) (~er cent V/V) (~er cent V/V) incubate at 3 7 ± 1 ºC for a further 2 h, protected from light.
o- 5 20 o 80 Add 10 µL of 2-mercaptoethanol R and mix. Dialyse against
1000 mL of solution C. Add 200 µL of solution C and mix.
5 - 21 20 0-74 80 -7 76
Determine the protein content of the solution.
21 - 61 20 4 -7 25 76 -7 55 Reference salutian (a). Prepare at the same time and in the
61 - 62 20 25 -7 50 55 -7 30 same manner as for the test solution but using follitropin
far peptide mapping and glycan analysis CRS instead of
62 - 71 20 50 30 the preparation to be examined. Determine the protein
71 - 72 20 50 -7 o 30 -7 80 content of the solution.
Reference salutian (b ). Prepare at the same time and in
72 - 117 20 o 80
the same manner as for the test solution but using fetuin
Flow rate: 0.4 mL/min. instead of the preparation to be examined. Determine the
protein content of the solution.
Detection: fluorimeter at 330 nm for excitation and at
420 nm for emission.
SELECTIVE RELEASE OF THE GLYCANS
Injection: SO µL. Test solutian. Dilute the test solution obtained in the
previous step with solution C to obtain a concentration of
System suitability: reference solution: 1.1 g/L. Add 1 U of peptide N-glycosidase F R to SOO µg of
- the chromatogram obtained is qualitatively similar to the solution, mix and incubate at 37 ± 1 ºC for 24 h. Place
the chromatogram supplied with follitropin for peptide the solution in ice. Precipitate the protein and salts with
mapping and glycan analysis CRS; 3 volumes of ice-cold anhydrous ethanal R and allow to
- by comparison with the chromatogram supplied with stand in ice for 10 min. Centrifuge at 16 000 g for about
f ollitropin f or peptide mapping and glycan analysis CRS, S min and transfer the supernatant to a separate tube. Add
identify the peaks due to neutral, mono-, di-, tri- and 3 µL of a 1 µg/µL solution of maltotriase R then freeze-dry.
tetra-sialylated forms; determine the area of each peak Dissolve in 100 µL of water for chromatagraphy R.
and express it as a percentage of the total; calculate the Reference salutian (a). Prepare in the same manner as for
Z number using the following expression: the test solution but using the reference solution obtained
with follitrapin far peptide mapping and glycan analysis CRS
in the previous step.
(Ao x O)+ (A1 x 1) + (A2 x 2) + (A3 x 3) + (A4 x 4)
Reference solution (b ). Prepare in the same manner as for
the test solution but using the reference solution obtained
A0 peak area percentage due to the neutral with fetuin in the previous step.
form;
CHROMATOGRAPHIC SEPARATION. Liquid
A1 peak area percentage due to the chromatography (2.2.29).
mono-sialylated form; Precolumn:
- size: l =O.OS m, 0 = 4.0 mm;
A2 peak area percentage dueto the di-sialylated - stationary phase: strongly basic anian-exchange resin for
form; chromatagraphy R.
Calumn:
A3 peak area percentage due to the
tri-sialylated form; - size: l = 0.2S m, 0 = 4.0 mm;
- stationary phase: strongly basic anian-exchange resin far
A4 peak area percentage due to the chromatagraphy R.
tetra-sialylated form. Mobile phase:
The Z number obtained for the reference solution is in - mobile phase A: 20 g/L solution of sodium hydraxide R;
the range 177-233. maintain under helium;
Examine the chromatogram obtained with the test solution - mobile phase B: water for chromatagraphy R; maintain
and calculate the Z number as described above. under helium;
Result: Z = 177-233. - mobile phase C: dissolve 41 g of anhydraus sadium
acetate R in 800 mL of water f or chromatography R,
METHOD B
dilute to 1000 mL with the same solvent, then mix; filter
PROTEIN DENATURATION through a membrane filter (nominal pore size 0.4S µm);
Solution A. To 1.9S2 g of 2-[N-morpholino]ethanesulfonic maintain under helium;
acid R and S7.32 g of guanidine hydrochloride R, add 1 mL Time Mobile phase A Mobile phase B Mobile phase C
of a lS.4 g/L solution of dithiothreitol R, 10 mL of an
(min) (~er cent V/V) (~er cent V/V) (~er cent V/V)
18.61 g/L solution of sodium edetate R and 20 mL of water
for chromatography R. Maintain in a water-bath at about
o - 0.2 20 80 o
37 ºC for 1 min to dissolve the components. Adjust to 0.2 - 94.0 20 80 -7 34 o -7 46
pH 8.1 (2.2.3) with an 80 g/L solution of sodium hydroxide R
94.0 - 97.0 20 34 46
and dilute to 100.0 mL with water for chromatography R.
Mix. 97.0 - 97.1 20 34 -7 80 46 -7 o
Solution B. Dissolve 37 mg of iodoacetamide R in 1 mL of 97.1 - 115.0 20 80 o
water for chromatography R and mix. Protect from light.
Solutian C. Dissolve 26.7 g of disadium hydragen phasphate Flaw rate: 1.0 mL/min.
dihydrate R and 11.2 g of sodium edetate R in 3000 mL of Detectian: pulsed amperometric detector or equivalent with
water for chromatagraphy R and mix. Adjust to pH 7.S a gold indicator electrode, a silver-silver chloride reference
(2.2.3) with a 40 g/L solution of sadium hydraxide R. electrode, and a stainless steel auxiliary electrode which
Test salution. To a volume of the preparation to be is the cell body, held at respectively + O.OS V detection,
examined that contains 1 mg of follitropin add 0.2 mL of + 0.7S V oxidation and - 0.80 V reduction potentials, with
solution A and incubate in a water-bath at 37 ± 1 ºC for pulse durations according to the instrument used.
S672 See the information section an general managraphs (caver pages)

Injection: 45 µL. System suitability: reference solution:

System suitability: - resolution: minimum 1.2 between the peaks due to bovine
- the chromatogram obtained with reference solution (b) albumin and follitropin;
is qualitatively similar to the chromatogram for fetuin - no peak is detected between 5 min and 16 min in blank
supplied with follitropin for peptide mapping and glycan injections.
analysis CRS; Limit:
the chromatograms obtained with the test solution - sum of the peaks with a retention time less than that of the
and reference solution (a) are qualitatively similar to principal peak: maximum 0.5 per cent.
the chromatogram supplied with follitropin for peptide
mapping and glycan analysis CRS; Free subunits. Polyacrylamide gel electrophoresis (2.2.31)
under non-reducing conditions.
by comparison with the chromatogram supplied with
follitropin for peptide mapping and glycan analysis CRS, Gel dimensions: 1.5 mm thick.
identify the peaks due to neutral, mono-, di-, tri- and Resolving gel: 12 per cent acrylamide.
tetra-sialylated forms in the chromatogram obtained Sample buffer. Concentrated SDS-PAGE sample buffer R.
with reference solution (b) ; determine the area of each Test solution. Dilute the preparation to be examined with
peak and express it as a percentage of the total; calculate water R to obtain a concentration of 2 µg/µL. To 55 µL of the
the Z number using the following expression: solution add 55 µL of the sample buffer. Allow to stand for 4 h
at room temperature.
(Ao X O) + (A1 X 1) + (A2 X 2) + (A3 X 3) + (A4 X 4) Reference solution (a). Concentrate follitropin CRS using a
suitably validated procedure to obtain a concentration of
2 µg/µL. To 25 µL of the solution add 25 µL of the sample
A0 peak area percentage due to the neutral
buffer. To 40 µL of this solution add 180 µL of the sample
form;
buffer and 180 µL of water R. Allow to stand for 4 h at room
A1 peak area percentage dueto the temperature, then boil for 5 min.
mono-sialylated form; Reference solution (b ). A solution of molecular mass markers
suitable for calibrating SDS-polyacrylamide gels in the range
A2 peak area percentage dueto the di-sialylated of 14.4-94 kDa.
form; Application :
A3 peak area percentage due to the Well Solution(s) Volume (µL)

tri-sialylated form; Reference solution (a) 40

A4 peak area percentage due to the
tetra-sialylated form. Reference solution (a) 20
The Z number obtained for reference solution (b) is in 4 Reference solution (a) 15
the range 290-325. Reference solution (a) 10
Examine the chromatogram obtained with the test solution
6 Reference solution (a)
and calculate the Z number as described above.
Result: Z = 178-274. 7 Test solution 50
TESTS 8 Test solution + reference solution (a) 50 + 25
Follitropin oligomers. Size-exclusion chromatography 9 Reference solution (b) 10

(2.2.30). Use the normalisation procedure.
Solution A. Dissolve 118 mg of sodium dihydrogen phosphate R, Detection: by Coomassie staining.
1.65 g of disodium hydrogen phosphate dihydrate R, and 30.0 g System suitability:
of sucrose R in 40 mL of water for chromatography R and - reference solution (b): the validation criteria are met
dilute to 100.0 mL with the same solvent. (2.2.31);
Solution B. Dissolve 2.0 mg of bovine albumin R in 30 mL of - test solution + reference solution (a): the bands
solution A. corresponding to the follitropin heterodimer and subunits
Test solution. Dilute the preparation to be examined with are clearly separated;
solution A to obtain a concentration of 0.25 mg/mL. - reference solution (a): no bands corresponding to the
Reference solution. Dilute follitropin CRS with solution A to follitropin heterodimer are seen.
obtain a concentration of 0.5 mg/mL and mix equal volumes Limit:
of this solution and solution B to obtain a concentration of - free subunits: maximum 3 per cent.
0.25 mg/mL.
Oxidised follitropin. Liquid chromatography (2.2.29).
Column:
Solution A. Dissolve about 3.3 mg of 2,4-dichlorobenzoic
- size: l = 0.3 m, 0 = 7.8 mm;
acid R in 10.0 mL of ethanol (96 per cent) R.
- stationary phase: hydrophilic silica gel for chromatography R,
of a grade suitable for fractionation of globular proteins
Test solution. Dilute the preparation to be examined in water
in the relative molecular mass range of 10 000 to 500 000
for chromatography R to obtain a concentration of 300 µg/mL.
(5 µm). Reference solution (a). Dilute follitropin CRS with water for
Mobile phase: dissolve 28.4 g of anhydrous sodium sulfate R in chromatography R to obtain a concentration of 300 µg/mL.
2000 mL of 0.1 M phosphate buffer solution pH 6.7 R and filter Reference solution (b ). Dilute 0.1 mL of strong hydrogen
through a membrane filter (nominal pore size 0.45 µm). peroxide solution R to 30 mL with water for chromatography R.
Flow rate: 0.5 mL/min. Dilute follitropin CRS with this solution to obtain a
concentration of 300 µg/mL. Incubate for 30-45 min. Add
Detection: spectrophotometer at 215 nm. solution A to obtain a concentration in 2,4-dichlorobenzoic
Injection: 100 µL. acid of about 17 µg/mL in the total volume and inject
Retention time: follitropin = 14-16 min. immediately.
Column: Mobile phase: mix 6.74 mL of phosphoric acid R, 14.2 g

- size: l = 0.25 m, 0 = 4.6 mm; of anhydrous sodium sulfate R and 900 mL of water for
chromatography R, adjust to pH 6.7 (2.2.3) with a 0.5 g/mL
- stationary phase: end-capped butylsilyl silica gel for solution of sodium hydroxide R and dilute to 1000 mL with
chromatography R (5 µm); water for chromatography R; filter through a membrane filter
- temperature: 30 ºC. (nominal pore size 0.45 µm).
Mobile phase: Flow rate: 1 mL/min.
- mobile phase A: 0.2 M phosphate buffer solution pH 2.5 R; Detection: spectrophotometer at 214 nm.
- mobile phase B: water for chromatography R, acetonitrile Rl
(40:60 V/V); lnjection: 100 µL.
- mobile phase C: water for chromatography R; System suitability: reference solution:
Time Mobile phase A Mobile phase B Mobile phase C - number of theoretical plates: minimum 1300, calculated for
(min) (per cent V/V) (per cent V/V) (per cent V/V)
the peak due to follitropin.
o - 8.4 50 25 7 39 25 7 11 Calculate the content of follitropin taking into account the
assigned content of follitropin CRS.
8.4 - 8.5 50 39 7 45 11 7 5
Potency
8.5 - 15 50 45
The follicle-stimulating activity of follitropin is estimated by
15 - 15.l 50 45 7 25 5 7 25
comparing under given conditions its effect in enlarging the
15.l - 25 50 25 25 ovaries of immature rats treated with chorionic gonadotrophin
with the same effect of the International Standard preparation
Flow rate: 1.0 mL/min. of human recombinant follicle-stimulating hormone or of
Detection: spectrophotometer at 210 nm. a reference preparation calibrated in International Units.
The International Unit of FSH is the activity contained in
lnjection: 25 µL. stated amounts of the International Standard of human
System suitability: reference solution (b): recombinant follicle-stimulating hormone. The equivalence in
- the peaks dueto the oxidised follitropin a- and International Units of the International Standard is stated by
~-subunits are separated from the peaks due to the the World Health Organization.
non-oxidised follitropin subunits and from the peak due Use immature female rats of the same strain, 19-28 days old,
to 2,4-dichlorobenzoic acid; differing in age by not more than 3 days and having masses
- the chromatogram obtained is similar to the chromatogram such that the difference between the heaviest and the liehtest
supplied with follitropin CRS. rat is not more than 1O g. Assign the rats at random to 6equal
Calculate the percentage of oxidation of the follitropin groups of at least 5 rats. If sets of 6 litter mates are available,
assign 1 litter mate from each set to each group and mark
subunits using the following expression:
according to litter.
(A2 + A4) X 100
Choose 3 doses of the reference preparation and 3 doses of
A1 + A2 + A3 + A4 the preparation to be examined such that the smallest <lose
produces a positive response in sorne of the rats and the
A1 area of the peak dueto the follitropin a-subunit; largest <lose <loes not produce a maximal response in all of
A2 area of the peaks due to the oxidised follitropin the rats. Use doses in geometric progression and as an initial
a-subunit; approximation total doses of 1.5 IU, 3.0 IU and 6.0 IU may be
tried, although the <lose will depend on the sensitivity of the
A3 area of the peak due to the follitropin ~-subunit; rats used, which may vary widely.
A4 area of the peak due to the oxidised follitropin Dilute and dissolve respectively the total quantities of the
~-subunit.
preparation to be examined and of the reference preparation
Limit: corresponding to the daily doses to be used in sufficient
phosphate-albumin buffered saline pH 7.2 R such that the daily
- total oxidised forms: maximum 6 per cent.
<lose is administered in a volume of about 0.5 mL. The buffer
Bacteria! endotoxins (2.6.14): less than 0.1 IU per solution shall contain in the daily <lose not less than 14 IU of
International Unit of follitropin activity, if intended for use in chorionic gonadotrophin to ensure complete luteinisation.
the manufacture of parenteral preparations without a further Add a suitable antimicrobial preservative such as 4 g/L of
appropriate procedure for the removal of bacteria! endotoxins. phenol or 0.02 g/L of thiomersal. Store the solutions at
5 ± 3 ºC.
ASSAY
Inject subcutaneously into each rat the daily <lose allocated to
Protein. Size-exclusion chromatography (2.2.30). its group. Repeat the injection of each <lose 24 h and 48 h after
Solution A. Dissolve 100 mg of poloxamer 188 R in 900 mL of the 1st injection. About 24 h after the last injection, euthanise
water for chromatography R and dilute to 1000 mL with the the rats and remove the ovaries from each rat. Remove any
same solvent. extraneous fluid and tissue from the ovaries and weigh the
Test solution. Dilute the preparation to be examined with 2 combined ovaries of each rat immediately. Calculate the
solution A to obtain a concentration of about 0.03 mg/mL. results by the usual statistical methods (for example, 5.3),
using the mass of the 2 combined ovaries as the response. (The
Reference solution. Dilute follitropin CRS with solution A to
precision of the assay may be improved by a suitable correction
obtain a concentration of about 0.03 mg/mL.
of the organ mass with reference to the mass of the rat from
Column: which it was taken; an analysis of covariance may be used.)
- size: l = 0.3 m, 0 = 7.8 mm; The estimated potency is not less than 80 per cent and not
- stationary phase: hydrophilic silica gel for chromatography R, more than 125 per cent of the stated potency. The confidence
of a grade suitable for fractionation of globular proteins limits (P = 0.95) of the estimated potency are not less than
in the relative molecular mass range of 10 000 to 500 000 64 per cent and not more than 156 per cent of the stated
(5 µm). potency.

STORAGE
In an airtight container, at a temperature not exceeding
- 20 ºC.
LABELLING
The label states:
- the content of protein in milligrams per millilitre;
- the potency in International Units per milligram of protein;
- where applicable, that the substance is suitable for use in
the manufacture of parenteral preparations.

G
Gemfibrozil.. ............................................................................ 5679 Glucosamine sulfate potassium chloride ............................. 5681
Glucosamine hydrochloride .................................................. 5680 Glucosamine sulfate sodium chloride .................................. 5682

EUROPEAN PHARMACOPOEIA 9.5 Gemfibrozil
07/2018:1694 Identification of impurities: use the chromatogram

supplied with gemfibrozil for system suitability CRS and the
chromatogram obtained with reference solution (a) to identify
the peaks due to impurities C, D and E; use the chromatogram
obtained with reference solution (c) to identify the peak due
to impurity A.
GEMFIBROZIL
Relative retention with reference to gemfibrozil
(retention time = about 7 min): impurity A = about 0.4;
Gemfibrozilum impurity C = about 1.3; impurity D = about 1.5;
impurity E = about 1.7; impurity I = about 2.0;
impurity H = about 2.9.
System suitability: reference solution (a):
gemfibrozil and impurity e, and minimum 2.0 between the
C1sH22Ü3 M, 250.3 peaks due to impurities D and E.
[25812-30-0] Limits:
- correction f actors: for the calculation of content multiply the
DEFINITION
peak areas of the following impurities by the corresponding
5-(2,5-Dimethylphenoxy)-2,2-dimethylpentanoic acid. correction factor: impurity A = 0.5; impurity D = 1.8;
Content: 99.0 per cent to 101.0 per cent (anhydrous substance). impurity E = 0.2; impurity H = 0.6;
- impurities E, I: for each impurity, not more than twice the
CHARACTERS area of the principal peak in the chromatogram obtained
Appearance: white or almost white, waxy, crystalline powder. with reference solution (b) (0.2 per cent);
Solubility: practically insoluble in water, very soluble in - impurities A, D, H: for each impurity, not more than the
methylene chloride, freely soluble in anhydrous ethanol and area of the principal peak in the chromatogram obtained
in methanol. with reference solution (b) (O.l per cent);
- unspecified impurities: for each impurity, not more than the
IDENTIFICATION area of the principal peak in the chromatogram obtained
A. Melting point (2.2.14) : 58 ºC to 61 ºC. with reference solution (b) (0.10 per cent);
B. Infrared absorption spectrophotometry (2.2.24). - total: not more than 5 times the area of the principal peak
Comparison: gemfibrozil CRS. (0.5 per cent);
TESTS - disregard limit: 0.5 times the area of the principal peak in
Related substances. Liquid chromatography (2.2.29). (O.OS per cent).
Test solution. Dissolve 40.0 mg of the substance to be Water (2.5.12): maximum 0.25 per cent, determined on
examined in mobile phase A and dilute to 10.0 mL with
2.000 g.
mobile phase A.
Reference solution (a). Dissolve the contents of a vial of 2.0 g. Allow to stand for 1 h after the first moistening before
gemfibrozil for system suitability CRS (containing impurities
heating.
C, D and E) in 2.0 mL of acetonitrile R.
Reference solution (b ). Dilute 1.0 mL of the test solution to ASSAY
100.0 mL with mobile phase A. Dilute 1.0 mL of this solution Dissolve 0.200 g in 40 mL of methanol R. Add 10 mL of
to 10.0 mL with mobile phase A. water R and 1 mL of 0.1 M hydrochloric acid. Carry out
Reference solution (e). Dissolve 5 mg of 2,5-dimethylphenol R a potentiometric titration (2.2.20) using 0.1 M sodium
(impurity A) in mobile phase A and dilute to 1O.O mL with hydroxide. Read the volume added between the 2 points of
mobile phase A. inflexion.
Column: 1 mL of 0.1 M sodium hydroxide is equivalent to 25.03 mg
of C 15H 22 0 3 •
- size: l = 0.25 m, 0 = 4.0 mm;
- stationary phase: end-capped octadecylsilyl silica gel for STORAGE
chromatography R (5 µm). Protected from light.
Mobile phase:
IMPURITIES
- mobile phase A: dissolve 0.49 g of potassium acetate R in Specified impurities: A, D, E, H, J.
400 mL of water for chromatography R, adjust to pH 4.0
with phosphoric acid R and add 600 mL of acetonitrile R; Other detectable impurities (the following substances would,
- mobile phase B: acetonitrile R; the tests in the monograph. They are limited by the general
Time Mobile phase A Mobile phase B acceptance criterion for other/unspecified impurities and/or
(min) (per cent V/V) (per cent V/V) by the general monograph Substances for pharmaceutical use
o- 5 100 o (2034). It is therefore not necessary to identify these impurities
for demonstration of compliance. See also 5.10. Control of
5 - 20 100 -7 o o -7 100 impurities in substances for pharmaceutical use): B, C, F, G.
20 - 25 o 100
H3CyYOH

~CH3
Injection: 20 µL. A. 2,5-dimethylphenol (p-xylenol),
Glucosamine hydrochloride EUROPEAN PHARMACOPOEIA 9.5

PRODUCTION
The animals from which glucosamine hydrochloride is derived
must fulfil the requirements for the health of animals suitable
B. 5-(2,5-dimethylphenoxy)-2,2-dimethylpentanamide, for human consumption.
H3Cu0~0~./"--...
O CH3 CHARACTERS
1
~ CH3
Appearance: white or almost white, crystalline powder.
Solubility: freely soluble in water, slightly soluble in methanol,
C. 2-[3-(2-ethoxyethoxy)propoxy]-1,4-dimethylbenzene, practically insoluble in acetone.
/CH 3 IDENTIFICATION
H3C CH3 A. Infrared absorption spectrophotometry (2.2.24).
H3C
Ú O~
CH 3
C02H
D. 5-[3,6-dimethyl-2-(prop-1-en-l-yl)phenoxy]-2,2-
Comparison: glucosamine hydrochloride CRS.
B. 1 mL of solution S (see Tests) gives reaction (a) of chlorides
(2.3.1).
C. Specific optical rotation (see Tests).
dimethylpentanoic acid, TESTS
Solution S. Dissolve 2.50 g in carbon dioxide-free water R and
dilute to 25.0 mL with the same solvent.
Appearance of solution. The solution is clear (2.2.1) and
colourless (2.2.2, Method II).
E. 5-[2,5-dimethyl-4-(prop-1-en-1-yl)phenoxy]-2,2- Dilute 5.0 mL of solution S to 25.0 mL with water R.
dimethylpentanoic acid, pH (2.2.3): 3.0 to 5.0 for solution S.
H3CY'y0~ Specific optical rotation (2.2.7): + 70.0 to+ 74.0 (dried
substance), determined on solution S.
~CH3 V Examine 3 h after preparation of solution S.
F. 1,4-dimethyl-2-(4-phenylbutoxy)benzene, Related substances. Liquid chromatography (2.2.29).
Test solution. To 0.300 g of the substance to be examined add
H3Cu0~ 1 CH2
80 mL of the mobile phase and sonicate for 10 min. Cool to
room temperature and dilute to 100.0 mL with the mobile
~ CH3 phase.
G. l,4-dimethyl-2-(prop-2-en-1-yloxy)benzene,
Reference solution (a). Dissolve 25.0 mg of 2-methylpyra-
zine CRS in the mobile phase and dilute to 10.0 mL with the
H3CyryO~OyYCH3 mobile phase. Dilute 1.0 mL of the solution to 10.0 mL with
the mobile phase. Dilute 1.0 mL of this solution to 100.0 mL
~CH3 H3C,YJ with the mobile phase.
H. l, 1'-[propane-1,3-diylbis( oxy) ]bis(2,5-dimethylbenzene ),
Reference solution (b). Dissolve the contents of a vial of
glucosamine for system suitability CRS (containing impurities B
H3C CH3 and C) in 1.0 mL of the mobile phase.
H3cyyo~OCH3 Column:
~ CH 3
o - size: l = 0.15 m, 0 = 4.6 mm;
- stationary phase: base-deactivated end-capped octadecylsilyl
I. methyl 5-(2,5-dimethylphenoxy)-2,2-dimethylpentanoate. silica gel for chromatography R (3 µm);
- tempera tu re: 30 ºC.
07/2018:2446 Mobile phase: dissolve 0.5 g of sodium heptanesulfonate R
Bi in water for chromatography R, add 0.5 mL of phosphoric
~ acid R and 4 mL of a 56 g/L solution of potassium hydroxide R

and dilute to 1000 mL with water for chromatography R; to
1000 mL of this solution add 50 mL of acetonitrile Rl.
GLUCOSAMINE HYDROCHLORIDE Flow rate: 1.0 mL/min.
Glucosamini hydrochloridum Injection: 20 µL.
Run time: twice the retention time of 2-methylpyrazine.
Retention time: 2-methylpyrazine = about 9 min.
HO)-o, OH , HCI
H~ NH2
impurities B and C.
Limits:
C6 H 14ClN0 5 Mr 215.6 - unspecified impurities: for each impurity, not more than
[66-84-2] 0.5 times the area of the principal peak in the chromatogram
obtained with reference solution (a) (O.OS per cent);
DEFINITION - total: not more than twice the area of the principal peak
2-Amino-2-deoxy-D-glucopyranose hydrochloride. in the chromatogram obtained with reference solution (a)
Isolated from natural sources or produced by fermentation. (0.2 per cent);

EUROPEAN PHARMACOPOEIA 9.5 Glucosamine sulfate potassium chloride
- disregard limit: 0.3 times the area of the principal peak in 07/2018:2708
the chromatogram obtained with reference solution (a)
(0.03 per cent).
on 1.000 g by drying in an oven at 105 ºC for 2 h.
GLUCOSAMINE SULFATE POTASSIUM
1.0 g. CHLORIDE
Microbial contamination
Glucosamini sulfas kalii chloridum
TAMC: acceptance criterion 103 CFU/g (2.6.12).
TYMC: acceptance criterion 102 CFU/g (2.6.12).
Absence of Escherichia coli (2.6.13).
ASSAY
Dissolve 0.200 g in 50 mL of water R and add 1.0 mL of 0.1 M
hydrochloric acid. Titrate with 0.1 M sodium hydroxide, C 12 H 28 Cl2K2 N 20 14S Mr 606
determining the end-point potentiometrically (2.2.20). Read
the volume added between the 2 points of inflexion. DEFINITION
1 mL of 0.1 M sodium hydroxide is equivalent to 21.56 mg of Bis(2-amino-2-deoxy-D-glucopyranose) sulfate bis(potassium
C6 H 14ClN0 5 • chloride).
Substance prepared from glucosamine hydrochloride isolated
IMPURITIES from natural sources or produced by fermentation, and
potassium sulfate.
if present at a sufficient level, be detected by one or other of Content: 98.0 per cent to 102.0 per cent (dried substance).
the tests in the monograph. They are limited by the general PRODUCTION
by the general monograph Substances for pharmaceutical use The animals from which glucosamine sulfate potassium
(2034). It is therefore not necessary to identify these impurities chloride is derived must fulfil the requirements for the health
for demonstration of compliance. See also 5.1 O. Control of of animals suitable for human consumption.
impurities in substances for pharmaceutical use): A, B, C, E. CHARACTERS
Appearance: white or almost white, crystalline powder.
Solubility: freely soluble in water, sparingly soluble in
methanol, practically insoluble in acetone.
IDENTIFICATION
A. Specific optical rotation (see Tests).
B. Infrared absorption spectrophotometry (2.2.24).
Comparison: glucosamine sulfate potassium chloride CRS.
C. It gives reaction (a) of chlorides (2.3.1).
A. 2-( acetylamino )-2-deoxy-D-glucopyranose (N-acetyl- D. It gives reaction (a) of sulfates (2.3.1).
glucosamine ), E. 1 mL of solution S (see Tests) gives reaction (a) of potassium
(2.3.1).
TESTS
Solution S. Dissolve 2.50 gin carbon dioxide-free water R and
dilute to 25.0 mL with the same solvent.
Appearance of solution. The solution is clear (2.2.1) and
Dilute 5.0 mL of solution S to 25.0 mL with water R.
B. ( lR, 1'R,2S,2' S,3R,3' R)-1, 1'-pyrazine-2,5-diylbis(butane-
l ,2,3,4-tetrol) (fructosazine ), pH (2.2.3): 3.0 to 5.0 for solution S.
Specific optical rotation (2.2.7): + 47.0 to+ 53.0 (dried
Examine 3 h after preparation of solution S.
Test solution. To 0.42 g of the substance to be examined add
80 mL of the mobile phase and sonicate for 1O min. Cool to
C. ( 1R,2S,3R)-1-[5-[ (2S,3R)-2,3,4-trihydroxybutyl]pyrazin-2- phase.
yl ]butane-1,2,3,4-tetrol (deoxyfructosazine ), Reference solution (a). Dissolve 25.0 mg of 2-methylpyra-
zine CRS in the mobile phase and dilute to 10.0 mL with the
mobile phase. Dilute 1.0 mL of the solution to 10.0 mL with
u
OHCYOV"
OH
the mobile phase. Dilute 1.0 mL of this solution to 100.0 mL
with the mobile phase.
Reference solution (b ). Dissolve the contents of a vial of
E. 5-(hydroxymethyl)furan-2-carbaldehyde (5-hydroxy- glucosamine for system suitability CRS (containing impurities B
methylfurfural). and C) in 1.0 mL of the mobile phase.
Glucosamine sulfate sodium chloride EUROPEAN PHARMACOPOEIA 9.5
Column:
- size: l = 0.15 m, 0 = 4.6 mm;
silica gel for chromatography R (3 µm) (3 µm);
- temperature: 30 ºC. B. (lR,l' R,25,2' S,3R,3' R)-l,l '-pyrazine-2,5-diylbis(butane-
Mobile phase: dissolve 0.5 g of sodium heptanesulfonate R in l,2,3,4-tetrol) (fructosazine),
water for chromatography R, add 0.5 mL of phosphoric acid R, HO H H OH
("~º"
4 mL of a 56 g/L solution of potassium hydroxide R and dilute
to 1000 mL with water for chromatography R, then add 50 mL
of acetonitrile Rl.
"·1º"
HO~N
Flow rate: 1.0 mL/min. HO H
Detection: spectrophotometer at 195 nm. C. (1R,2S,3R)-1-[5-[ (2S,3R)-2,3,4-trihydroxybutyl]pyrazin-2-
Injection: 20 µL. yl] butane-1,2,3,4-tetrol (deoxyfructosazine ),
Run time: twice the retention time of 2-methylpyrazine. ~OyCHO
Retention time: 2-methylpyrazine = about 9 min. HO \_j/
E. 5-(hydroxymethyl)furan-2-carbaldehyde (5-hydroxy-
- resolution: minimum 1.5 between the peaks due to methylfurfural).
impurities B and C.
Calculation of percentage contents: 07/2018:2447
- for each impurity, use the concentration of2-methylpyrazine
in reference solution (a).
Limits:
- unspecified impurities: for each impurity, maximum GLUCOSAMINE SULFATE SODIUM
O.OS per cent;
CHLORIDE
- reporting threshold: 0.03 per cent. Glucosamini sulfas natrii chloridum
on 1.000 g by drying in an oven at 105 ºC for 2 h.
Sulfated ash (2.4.14): 27.0 per cent to 31.0 per cent,
determined on 1.0 g.
Microbial contamination
TAMC: acceptance criterion 103 CFU/g (2.6.12).
TYMC: acceptance criterion 102 CFU/g (2.6.12). Mr 573.3
Absence of Escherichia coli (2.6.13). DEFINITION
Bis(2-amino-2-deoxy-D-glucopyranose) sulfate bis(sodium
ASSAY
chloride).
Dissolve 0.280 g in 50 mL of water R and add 1.0 mL of 0.1 M Substance prepared from glucosamine hydrochloride isolated
hydrochloric acid. Titrate with 0.1 M sodium hydroxide, from natural sources or produced by fermentation, and
determining the end-point potentiometrically (2.2.20). Read sodium sulfate.
the volume added between the 2 points of inflexion.
1 mL of 0.1 M sodium hydroxide is equivalent to 30.28 mg
of C 12 H 28 Cl2 K2 N 2 0 14S. PRODUCTION
The animals from which glucosamine sulfate sodium chloride
IMPURITIES is derived must fulfil the requirements for the health of
Other detectable impurities (the following substances would, animals suitable for human consumption.
the tests in the monograph. They are limited by the general CHARACTERS
acceptance criterion for other/unspecified impurities and/or Appearance: white or almost white, crystalline powder.
by the general monograph Substances for pharmaceutical use Solubility: freely soluble in water, sparingly soluble in
(2034). It is therefore not necessary to identify these impurities methanol, practically insoluble in acetone.
impurities in substances for pharmaceutical use): A, B, C, E. IDENTIFICATION
Comparison: glucosamine sulfate sodium chloride CRS.
B. It gives reaction (a) of chlorides (2.3.1).
HO)-o, OH
C. 1 mL of solution S (see Tests) gives reaction (a) of sodium
H~ (2.3.1).
D. It gives reaction (a) of sulfates (2.3.1).
HNYCH3
E. Specific optical rotation (see Tests).
o
TESTS
A. 2-(acetylamino)-2-deoxy-D-glucopyranose (N-acetyl- Solution S. Dissolve 2.50 g in carbon dioxide-free water R and
glucosamine), dilute to 25.0 mL with the same solvent.

EUROPEAN PHARMACOPOEIA 9.S Glucosamine sulfate sodium chloride
Appearance of solution. The solution is clear (2.2.1) and Microbial contamination

colourless (2.2.2, Method JI). TAMC: acceptance criterion 103 CFU/g (2.6.12).
Dilute S.O mL of solution S to 2S.O mL with water R. TYMC: acceptance criterion 10 2 CFU/g (2.6.12).
pH (2.2.3): 3.0 to S.0 for solution S. Absence of Escherichia coli (2.6.13).
Specific optical rotation (2.2.7): + SO.O to + SS.O (dried
substance), determined on solution S. ASSAY
Examine 3 h after preparation of solution S. Dissolve 0.2SO g in SO mL of water R and add 1.0 mL of 0.1 M
hydrochloric acid. Titrate with 0.1 M sodium hydroxide,
Related substances. Liquid chromatography (2.2.29). determining the end-point potentiometrically (2.2.20). Read
Test solution. To 0.400 g of the substance to be examined add the volume added between the 2 points of inflexion.
80 mL of the mobile phase and sonicate for 10 min. Cool to
1 mL of 0.1 M sodium hydroxide is equivalent to 28.67 mg
of C 12H 28 Cl2 N 2 Na2 0 14S.
phase.
Reference solution (a). Dissolve 2S.O mg of 2-methylpyra- IMPURITIES
zine CRS in the mobile phase and dilute to 10.0 mL with the Other detectable impurities (the following substances would,
mobile phase. Dilute 1.0 mL of the solution to 10.0 mL with if present at a sufficient level, be detected by one or other of
the mobile phase. Dilute 1.0 mL of this solution to 100.0 mL the tests in the monograph. They are limited by the general
with the mobile phase. acceptance criterion for other/unspecified impurities and/or
Reference solution (b ). Dissolve the contents of a vial of by the general monograph Substances for pharmaceutical use
glucosamine for system suitability CRS (containing impurities B (2034). It is therefore not necessary to identify these impurities
and C) in 1.0 mL of the mobile phase. for demonstration of compliance. See also 5.1 O. Control of
Column: impurities in substances for pharmaceutical use): A, B, C, E.
- size: l = 0.15 m, 0 = 4.6 mm;
silica gel for chromatography R (3 µm);
Mobile phase: dissolve 0.5 g of sodium heptanesulfonate R
in water for chromatography R, add 0.5 mL of phosphoric
acid R and 4 mL of a S6 g/L solution of potassium hydroxide R
and dilute to 1000 mL with water for chromatography R; to
1000 mL of this solution add SO mL of acetonitrile Rl. A. 2-(acetylamino)-2-deoxy-D-glucopyranose (N-acetyl-
Flow rate: 1.0 mL/min. glucosamine ),
Detection: spectrophotometer at 19S nm.
Injection: 20 µL.
Run time: twice the retention time of 2-methylpyrazine.
Retention time: 2-methylpyrazine = about 9 min.
- resolution: minimum l.S between the peaks due to
impurities B and C. B. (IR, 1'R,25,2' S,3R,3'R)-1,1 '-pyrazine-2,S-diylbis(butane-
1,2,3,4-tetrol) (fructosazine),
Limits:
- unspecified impurities: for each impurity, not more than
O.S times the area of the principal peak in the chromatogram
obtained with reference solution (a) (O.OS per cent);
- total: not more than twice the area of the principal peak
in the chromatogram obtained with reference solution (a)
(0.2 per cent);
- disregard limit: 0.3 times the area of the principal peak in C. (1R,2S,3R)-1-[S-[ (2S,3R)-2,3,4-trihydroxybutyl]pyrazin-2-
the chromatogram obtained with reference solution (a) yl ]butane-1 ,2,3,4-tetrol (deoxyfructosazine ),
(0.03 per cent).
~OyCHO
Loss on drying (2.2.32): maximum 0.5 per cent, determined HO 'L_j/
on 1.000 g by drying in an oven at lOS ºC for 2 h.
Sulfated ash (2.4.14): 23.5 per cent to 26.0 per cent, E. S-(hydroxymethyl)furan-2-carbaldehyde (S-hydroxy-
determined on 1.0 g. methylfurfural).

H
Human coagulation factor VIIa (rDNA) concentrated Hydroxyzine hydrochloride ................................................... 5693
solution .................................................................................. 5687 Hyoscine butylbromide.......................................................... 5694
Hydroxypropylcellulose, low-substituted ............................ 5691

EUROPEAN PHARMACOPOEIA 9.5 Human coagulation factor Vlla (rDNA) concentrated solution
01/2015:2534 Glycan analysis.

corrected 9.5 Use a suitable method developed according to general chapter
2.2.59. Glycan analysis of glycoproteins.
Glycan analysis includes the following steps:
- after desalting, release of the glycans (see 2.2.59 section 2-3);
HUMAN COAGULATION FACTOR Vlla - labelling of the glycans with a suitable fluorescent label
(Table 2.2.59.-2);
(rDNA) CONCENTRATED SOLUTION - analysis of the labelled glycans by liquid chromatography
(2.2.29) with fluorometric detection.
Factoris VIIa coagulationis humani (ADNr) The following procedures may be used.
solutio concentrata Test solution. Dilute the preparation to be examined in water R
light chain to obtain a concentration of about 1.5 mg/mL.
ANAFLEELRP GSL~R~CKg QCSFgAR~I FKDAERTKLF 40 Reference solution. Dissolve human coagulation factor VIIa
WISYSDGDQC ASSPCQNGGS CKQQLQSYIC FCLPAFEGRN 80 (rDNA) CRS in water R to obtain a concentration of
CETHKDDQLI CVNENGGCEQ YCSDHTGTKR SCRCHEGYSL 120 1.5 mg/mL.
LADGVSCTPT VEYPCGKIPI LEKRNASKPQ GR 152
DESALTING
heavy chain
IVGGKVCP 160
Desalt the test solution and the reference solution as described
KGECPWQVLL LVNGAQLCGG TLINTIWVVS AAHCFDKIKN 200
under Identification B. The buffer used for desalting and elution
WRNLIAVLGE HDLSEHDGDE QSRRVAQVII PSTYVPGTTN 240
is a 1.21 g/L solution of tris(hydroxymethyl)aminomethane R,
HDIALLRLHQ PVVLTDHVVP LCLPERTFSE RTLAFVRFSL 280
adjusted to pH 7.5 with hydrochloric acid R. After desalting,
VSGWGQLLDR GATALELMVL NVPRLMTQDC LQQSRKVGDS 320
the concentration of the solutions is about 1.0 mg/mL.
PNITEYMFCA GYSDGSKDSC KGDSGGPHAT HYRGTWYLTG 360 SELECTIVE RELEASE OF GLYCANS
IVSWGQGCAT VGHFGVYTRV SQYIEWLQKL MRSEPRPGVL 400 Transfer 500 µL of the desalted test solution and 500 µL of the
LRAPFP 406
desalted reference solution to separate centrifuge tubes, and
add 1O µL of a 200 U /mL solution of peptide N-glycosidase FR.
disulfide bridges:
17-22, 50-61, 55-70, 72-81, 91-102, 98-112, 114-127, 135-262, 159-164, Cap the tubes and incubate for 16-24 h at 37 ºC. Remove the
178-194, 310-329, 340-368 protein fraction by adding 1.5 mL of ethanol (96 per cent) R
glycosylation siles: at - 20 ºC to the tubes. Mix and allow to stand at - 20 ºC for
52, 60, 145, 322
20-30 min. Centrifuge the tubes at 10 000 r/min for 10 min.
modified residues: Collect the supernatant and evaporate to dryness, using for
f(4-carboxyGlu) at position 6, 7, 14, 16, 19, 20, 25, 26, 29, 35
example a rotary evaporator.
potentially modified residue:
Q ((3R)-3-hydroxyAsp) at position 63 LABELLING OF GLYCANS
H NH2 Label the liberated glycans with 2-aminobenzamide using a
H02C~
)\
X "C02H suitable procedure. The procedure employs a combination of
reagents optimised and validated for the efficient labelling of
HO H P"lvc::ins. and for the subseauent extraction and recoverv of the
f = 4-carboxyGlu Q = (3R)-3-hydroxyAsp f¡b~Ü-~d gly~¡~~ f~~m the :eaction. '
Mr approx. 50 000 LIQUID CHROMATOGRAPHY (2.2.29)
DEFINITION Precolumn:
Solution containing closely related glycoproteins, which have - size: l = 0.05m, 0 = 4.0 mm;
the same amino acid sequence (406 amino acids) and disulfide - stationary phase: strongly basic anion-exchange resin for
bridges as the naturally occurring analogue (plasma-derived chromatographyR.
activated coagulation factor VII). Human coagulation Column:
factor Vlla (rDNA) (eptacog alfa, activated) is a 2-chain - size: l = 0.25 m, 0 = 4.0 mm;
molecule, obtained by proteolytic cleavage of the peptide
bond between Arg 152 and lle 153 of single-chain coagulation - stationary phase: strongly basic anion-exchange resin for
factor VII, consisting of a 20 kDa light chain (N-terminal) and chromatography R;
a 30 kDa heavy chain (C-terminal) connected by a disulfide - temperature: 30 ºC.
bond. Mobile phase:
Human coagulation factor Vlla (rDNA) is distinguishable - mobile phase A: 6 g/L solution of sodium hydroxide R;
from the naturally occurring analogue in terms of its - mobile phase B: solution containing 6 g/L of sodium
post-translational modifications, including glycosylation hydroxide R and 40.8 g/L of sodium acetate R;
pattern.
Content: 1.11 mg to 1.78 mg of protein per millilitre.
Potency: 44 000 IU to 64 000 IU per milligram of protein. o - 52 100 7 35 o7 65
PRODUCTION 52.0 - 52.1 35 7 o 65 7 100

Human coagulation factor Vlla (rDNA) is produced in 52.1 - 65 o 100
mammalian cells by a method based on recombinant DNA
technology (rDNA). 65 - 65.1 o7 100 100 7 o
Prior to release, the following tests are carried out on each batch 65.1 - 90 100 o
of the final bulk product, unless exemption has been granted by
the competent authority. Flow rate: 0.5 mL/min.
Host-cell-derived proteins. The limit is approved by the Detection: fluorimeter at 330 nm for excitation and 420 nm
competent authority. for emission.
Host-cell- and vector-derived DNA. The limit is approved Injection: 100 µL, using an automatic injector maintained at
by the competent authority. 2-8 ºC.
Human coagulation factor Vlla (rDNA) concentrated solution EUROPEAN PHARMACOPOEIA 9.5
System suitability: reference solution: SELECTIVE CLEA VAGE OF THE PEPTIDE BONDS
- the chromatogram obtained is similar to the chromatogram Solution A. Dissolve 0.74 g of calcium chloride R and 6.06 g
shown in Figure 2534.-1; peaks 1 to 12 are clearly visible; of tris(hydroxymethyl)aminomethane R in 1000 mL of
- peak width at half-height: maximum 30 s for peak 8. water R and adjust to pH 7.5 with hydrochloric acid R.
Calculate the percentage content of charged glycans in the Test solution. Dilute the preparation to be examined with
reference solution using the following expression: solution A to obtain a concentration of about 1.5 mg/mL.
A Desalt a volume of this solution by a suitable method
A+B X 100 (for example using a suitable centrifuga! filter unit
or gel-filtration column with solution A as elution
A sum of the areas of the peaks due to charged buffer). After desalting, the concentration should be
glycans (peaks 6 to 12); about 1.0 mg/mL. Transfer the desalted solution to a
polypropylene tube. Prepare a 1 mg/mL solution of
B sum of the areas of the peaks due to uncharged
trypsin for peptide mapping R and add 10 µL to 1 mL of
glycans (peaks 1 to 5).
the desalted solution. Cap the tube and mix gently by
Calculate the percentage content of charged glycans in the inversion. Incubate at 37 ºC for 24 h. At time 5.5 h, add
test solution accordingly. 1O µL of the trypsin solution. Remove the sample from the
Limit: incubator, place it at room temperature, add 9 µL of glacial
acetic acid R and mix by inversion. Maintain the solution
- percentage of charged glycans: as authorised by the at - 15 ºC or below until chromatographic separation; if
competent authority. analysed immediately using an automatic injector, maintain
CHARACTERS at 2-8 ºC.
Appearance: colourless liquid. Reference solution. Dissolve human coagulation factor VIIa
(rDNA) CRS in solution A to obtain a concentration of
IDENTIFICATION 1.5 mg/mL. Desalt and digest at the same time and in the
A. It forms a clot when examined in the conditions described same manner as for the test solution.
under Assay (Potency). CHROMATOGRAPHIC SEPARA TION. Liquid
B. Peptide mapping (2.2.55). chromatography (2.2.29).
6 7 8
11
12
2
10
10 15 25 30 35 40 45 min
Peak Charged Structure Peak Charged Structure
l. No Core fucosylated biantennary - non sialylated 7. Yes Core fucosylated biantennary -

(2 N-acetylgalactosamine terminals) monosialylated (and 1 galactose
terminal)
2. No Core fucosylated biantennary - non sialylated 8. Yes Core fucosylated biantennary -
(N-acetylgalactosamine and galactose terminals) bisialylated
3. No Structure not determined 9. Yes High-mannose structure with
1 phosphate group
4. No Core fucosylated biantennary - non sialylated 10. Yes Core fucosylated triantennary -
(galactose and N-acetylglucosamine terminals) trisialylated
5. No Core fucosylated biantennary - non sialylated 11. Yes Core fucosylated triantennary -
(2 galactose terminals) trisialylated
6. Yes Core fucosylated biantennary - monosialylated 12. Yes Structure not determined
(and 1 N-acetylgalactosamine terminal)
Figure 2534.-1. - Chromatogram for the test for glycan analysis of human coagulation factor VIIa (rDNA): reference solution

EUROPEAN PHARMACOPOEIA 9.5 Human coagulation factor Vlla (rDNA) concentrated solution
Column: Time Mobile phase A Mobile phase B

(min) (per cent V/V) (per cent V!V)
size: l = 0.25 m, 0 = 4.0 mm;
o - 30 54 -7 41 46 -7 59
- stationary phase: octadecylsilyl silica gel far
chromatography R (5 µm) with a pore size of 30 nm; 30 - 33 41 -7 o 59 -7 100
- temperature: 30 ºC. 33 - 38 o 100
Mobile phase: 38 - 40 o -7 54 100 -7 46
- mobile phase A: add 0.65 mL of trifluoroacetic acid R to Flow rate: 1.0 mL/min.
1000 mL of water far chromatography R and <legas; Detection: spectrophotometer at 214 nm.
- mobile phase B: mix 0.5 mL of trifluoroacetic acid R, Injection: about 20 µL, using an automatic injector maintained
100 mL of water for chromatography R and 900 mL of at 2-8 ºC.
acetonitrile Rl and <legas; Retention time: human coagulation factor VIIa (rDNA) = about
Time Mobile phase A Mobile phase B 26 min.
(min) (per cent V/V) (per cent V/V) System suitability:
o - 100 100 -7 50 o -7 50 - the chromatogram obtained with the reference solution
is similar to the chromatogram shown in Figure 2534.-2;
100 - 105 50 -7 o 50 -7 100
peaks 1 to 1O are clearly visible;
105 - 110 o 100 - peak-to-valley ratio: minimum 1.5, where HP = height above
110 - 110.l o -7 100 100 -7 o the baseline of peak 6 and Hv = height above the baseline
of the lowest point of the curve separating this peak from
110. l - 125 100 o peak 7.
Results:
- the chromatogram obtained with the test solution is similar
Detection: spectrophotometer at 215 nm. to the chromatogram obtained with the reference solution.
Injection: 25 µL. Calculate the individual percentage area (relative to the total
peak area) of the peaks due to the degraded heavy chain
System suitability: the chromatogram obtained with the
human coagulation factor VIIa (rDNA) (peaks l, 2 and 6) and
reference solution is similar to the chromatogram supplied
oxidised forms of human coagulation factor VIIa (rDNA)
with human coagulation factor VIIa (rDNA) CRS.
(peaks 3, 4 and 5).
Results: the chromatogram obtained with the test solution Limits:
is similar to the chromatogram obtained with the reference
solution: - sum of degraded heavy chain forms: maximum 11 per cent;
- sum of oxidised forms: maximum 2.2 per cent.
- all major peaks identified in the chromatogram
obtained with the reference solution are present in the Gla-domainless human coagulation factor Vlla (rDNA)
chromatogram obtained with the test solution; (gamma-carboxylation). Liquid chromatography (2.2.29):
use the normalisation procedure.
- no new major peaks are observed in the chromatogram
Test solution. Dilute the preparation to be examined in water R
obtained with the test solution in comparison with the
to obtain a concentration of about 1.5 mg/mL.
chromatogram obtained with the reference solution.
Reference solution. Dissolve human coagulation factor VIIa
C. Examine the chromatograms obtained in the test for glycan (rDNA) CRS in water R to obtain a concentration of
analysis. 1.5 mg/mL.
Results: the chromatogram obtained with the test solution Precolumn:
is similar to the chromatogram obtained with the reference - stationary phase: styrene-divinylbenzene copolymer R with
solution. iminodiacetic groups, for removal of calcium.
Column:
TESTS
- size: l = 0.25 m, 0 = 4.0 mm;
Degraded heavy chain and oxidised forms of human - stationary phase: strongly basic anion-exchange resin far
coagulation factor Vlla (rDNA). Liquid chromatography chromatography Rl;
(2.2.29) : use the normalisation procedure.
Test solution. Dilute the preparation to be examined in water R Mobile phase:
to obtain a concentration of about 1.5 mg/mL. _ mobile phase A: solution containing 1.2 g/L of
Reference solution. Dissolve human coagulation factor VIIa tris(hydroxymethyl)aminomethane R and 2.8 g/L of bis-tris
(rDNA) CRS in water R to obtain a concentration of propane R, adjusted to pH 9.4 with glacial acetic acid R and
1.5 mg/mL. degassed;
Column: mobile phase B: solution containing 1.2 g/L of
tris(hydroxymethyl)aminomethane R, 2.8 g/L of bis-tris
- size: l = 0.25 m, 0 = 4.0 mm; propane R and 107.9 g/L of ammonium acetate R, adjusted
- stationary phase: end-capped butylsilyl silica gel for to pH 9.4 with concentrated ammonia R and degassed;
chromatography R (5 µm) with a pore size of 30 nm; Time Mobile phase A Mobile phase B
- temperature: 60-70 ºC. (min) (per cent V/V) (per cent V/V)
Mobile phase: o - 2.5 100 o
- mobile phase A: mix 1 mL of trifluoroacetic acid R and 2.5 - 27.5 100 -7 o o -7 100
999 mL of water for chromatography R and <legas; 27.5 - 30.5 o -7 100 100 -7 o
- mobile phase B: mix 1 mL of trifluoroacetic acid R, 200 mL
of water for chromatography R and 800 mL of acetonitrile Rl Flow rate: 1.0 mL/min.
and <legas; Detection: spectrophotometer at 280 nm.
Human coagulation factor Vlla (rDNA) concentrated solution EUROPEAN PHARMACOPOEIA 9.5
1 1 1 1 1 1 1
o 5 10 15 20 25 30 min
l. degraded heavy chain 3. oxidised form 5. oxidised form 7. human coagulation factor Vlla 9. unknown
(rDNA)
2. degraded heavy chain 4. oxidised form 6. degraded heavy chain 8. unknown 10. unknown
Figure 2534.-2. - Chromatogram for the test for degraded heavy chain and oxidised forms of human coagulation factor VIIa
(rDNA): reference solution
Injection: about 100 µL, using an automatic injector Mobile phase. Dissolve 26.4 g of ammonium sulfate R in
maintained at 2-8 ºC. approximately 900 mL of water f or chromatography R. Adjust
Relative retention with reference to human coagulation first to pH 2.5 with phosphoric acid R and then to pH 7.0 with
factor Vlla (rDNA) (retention time= about 14 min): triethylamine R. Add 50 mL of 2-propanol R and dilute to
Gla-domainless human coagulation factor Vlla 1000 mL with water for chromatography R.
(rDNA) = about 0.7. Flow rate: 0.5 mL/min.
System suitability: reference solution: Detection: spectrophotometer at 215 nm.
Injection: 20 µL, using an automatic injector maintained at
- resolution: baseline separation between the peak due to
2-8 ºC.
Gla-domainless human coagulation factor Vlla (rDNA)
and the peak cluster due to human coagulation factor Vlla System suitability: reference solution:
(rDNA). - the chromatogram obtained is similar to the chromatogram
Limit: supplied with human coagulationfactor VIIa (rDNA) CRS;
- symmetry factor: maximum 1.3 for the peak due to the
- Gla-domainless human coagulation factor VIIa (rDNA): monomer;
maximum 6.1 per cent.
- peak-to-valley ratio: minimum 1.1, where HP = height above
Integrate the peak cluster to baseline. the baseline of the peak due to dimers and Hv = height
above the baseline of the lowest point of the curve
separating this peak from the peak due to the monomer.
Dimers and related substances of higher molecular mass.
Size-exclusion chromatography (2.2.30): use the normalisation Limit:
procedure. - sum of the areas of the peaks with a retention time less than
Test solution. Dilute the preparation to be examined in water R that of the monomer: maximum 2.7 per cent.
to obtain a concentration of about 1.5 mg/mL. Non-activated single-chain factor VII (rDNA).
Reference solution. Dissolve human coagulation factor VIIa Polyacrylamide gel electrophoresis (2.2.31): use the
(rDNA) CRS in water R to obtain a concentration of normalisation procedure.
1.5 mg/mL. Gel dimensions: 1 mm thick.
Column: Resolving gel: 12 per cent acrylamide.
Sample buffer (reducing conditions): concentrated SDS-PAGE
- size: l = 0.3 m, 0 = 7.5 mm;
sample buffer for reducing conditions R containing
- stationary phase: hydrophilic silica gel for chromatography R dithiothreitol R as the reducing agent.
(10 µm) of a grade suitable for fractionation of globular Test solution. Dilute the preparation to be examined in
proteins in the relative molecular mass range of 1O 000 to water R to obtain a concentration of about 800 µg/mL. Mix
500 000; equal volumes of this solution and the sample buffer (reducing
- temperature: 21-25 ºC. conditions).

EUROPEAN PHARMACOPOEIA 9.5 Hydroxypropykellulose, low-substituted
Reference solution (a). Dissolve human coagulation factor VIIaPrepare 3 different solutions of the preparation to be
( rDNA) CRS in water R to obtain a concentration of about examined and of the reference preparation, by diluting with
800 µg/mL. Mix equal volumes of this solution and the sample solution A, to obtain concentrations within the linearity range
buffer (reducing conditions). (0.002-0.15 IU/mL). Prepare in duplicate and use the solutions
Reference solution (b). Solution of molecular mass markers immediately.
suitable for calibrating SDS-polyacrylamide gels in the range To 40 µL of each solution, add 40 µL of factor VII-deficient
of 10-70 kDa. plasma R, incubate for an appropriate time at 3 7 ºC, and add
Sample treatment: boil for 5 min or heat at 73 ± 3 ºC for 40 µL of human tissue factor solution R.
10 min. Measure the coagulation time, i.e. the interval between the
Application: 1O µL. addition of the human tissue factor solution and the first
Detection: by Coomassie staining. indication of the formation of fibrin.
Quantification: integrating densitometer. The volumes given above and sequence of reagents may be
System suitability: adapted to the human tissue factor solution and apparatus used.
- the principal bands in the electropherogram obtained with Calculate the activity in International Units per millilitre using
the test solution correspond in position to the principal an appropriate statistical method, for example the parallel-line
bands in the electropherogram obtained with reference assay (5.3).
solution (a) (30 kDa, heavy chain and 20-25 kDa, light The confidence limits (P = 0.95) are not less than 80 per cent
chain); and not more than 125 per cent of the estimated potency.
- reference solution (b): the validation criteria are met
(2.2.31);
LABELLING
- a band corresponding to non-activated single-chain The label states:
factor VII (rDNA) (molecular mass of 51 kDa) is visible in - the content of human coagulation factor Vlla (rDNA), in
the electropherogram obtained with reference solution (a). milligrams per millilitre;
Limit: - the specific activity, in International Units per milligram
- non-activated single-chain factor VII ( rDNA): maximum of protein.
3 per cent.
Bacteria! endotoxins (2. 6.14) : less than 1O IUI mL.
07/2018:2083
ASSAY
Protein. Size-exclusion chromatography (2.2.30) as described
in the test for dimers and related substances of higher
molecular mass with the following modifications.
Injection: 10 µL, 20 µL and 30 µL of the reference solution. HYDROXYPROPYLCELLULOSE,
Plot peak areas against injected protein content and perform a
linear regression to create a standard curve. LOW-SUBSTITUTED(l)
Calculate the content of human coagulation factor Vlla
(rDNA) using the monomer peak area in the chromatogram Hydroxypropylcellulosum
obtained with the test solution and taking into account substitutum humile
the assigned content of human coagulation factor VIIa
(rDNA) CRS.
[9004-64-2]
System suitability:
- repeatability: maximum relative standard deviation of DEFINITION
2.0 per cent after 5 injections of 20 µL of the reference Low-substituted 0-(2-hydroxypropylated) cellulose.
solution;
Content: 5.0 per cent to 16.0 per cent of hydroxypropoxy
- the correlation coefficient calculated for the standard curve groups (dried substance).
(r2) is not less than 0.990.
Potency. +CHARACTERS
The principle of the assay is to measure the ability of a Appearance: white or yellowish-white, hygroscopic powder or
factor Vlla preparation to reduce the prolonged coagulation granules.
time of factor VII-deficient plasma. Solubility: practically insoluble in ethanol (96 per cent). It
The biological activity is assessed by comparing the dissolves in a dilute solution of sodium hydroxide producing
<lose-response curve of the preparation to be examined to that a viscous solution. It swells in water, in a 106 g/L solution of
of a reference preparation calibrated in International Units. sodium carbonate and in a 206 g/L solution of hydrochloric
The International Unit is the activity contained in a stated acid R.+
amount of the International Reference Preparation.
IDENTIFICATION
The equivalence in International Units of the International
Reference Preparation is stated by the World Health A. Infrared absorption spectrophotometry (2.2.24).
Organization. Comparison: low-substituted hydroxypropylcellulose CRS.
Method. B. Shake thoroughly 0.1 g with 10 mL of water R. It <loes not
Use a suitable coagulation analyser or carry out the assay with dissolve.
incubation tubes and reagents maintained in a water-bath at C. To the suspension obtained in Identification B add
37 ºC. 1 g of sodium hydroxide R and shake until it becomes
Solution A. Prepare a solution containing 15.12 g/L of homogeneous. Transfer 5 mL of the solution to a suitable
1,4-piperazinediethanesulfonic acid R, 5.73 g/L of sodium container, add 1O mL of a mixture of 1 volume of
chloride R, 0.74 g/L of sodium edetate R and 10 g/L of bovine methanol R and 4 volumes of acetone R and shake; a white,
albumin R; adjust to pH 7.2 with sodium hydroxide R. flocculent precipitate is formed.
(1) This monograph has undergone pharmacopoeial harmonisation. See chapter 5.8. Pharmacopoeial harmonisation.
Hydroxypropylcellulose, low-substituted EUROPEAN PHARMACOPOEIA 9.5
TESTS Detection: flame ionisation or thermal conductivity.

pH (2.2.3): 5.0 to 7.5. Injection: 1-2 µL of the test solution and the reference solution.
Evenly distribute 1.0 g onto the surface of 100 mL of carbon
dioxide-free water R and stir using a magnetic stirrer. Relative retention with reference to octane (retention
time = about 8 min): isopropyl iodide = about 0.8.
on 1.000 g by drying in an oven at 105 ºC for 1 h. System suitability: reference solution:
Sulfated ash (2.4.14): not more than 0.8 per cent, determined - resolution: mínimum 5.0 between the peaks due to
on 1.0 g. isopropyl iodide and octane;
ASSAY - repeatability: maximum relative standard deviation of

2.0 per cent for the ratio of the area of the peak due to
Gas chromatography (2.2.28). isopropyl iodide to that due to octane determined on
Apparatus: 6 injections.
- reaction vial: a 5 mL pressure-tight vial, 50 mm in height, Calculate the ratio ( Q) of the area of the peak due to isopropyl
20 mm in externa! diameter and 13 mm in interna! diameter iodide to the area of the peak due to the internal standard
at the mouth, equipped with a pressure-tight butyl rubber from the chromatogram obtained with the test solution, and
membrane stopper coated with polytetrafluoroethylene and the ratio ( Q 1) of the area of the peak due to isopropyl iodide
secured with an aluminium crimped cap or another sealing to the area of the peak due to the interna! standard from the
system providing a sufficient air-tightness; chromatogram obtained with the reference solution.
heater: a heating module with a square aluminium block Calculate the percentage content of hydroxypropoxy groups
having holes 20 mm in diameter and 32 mm in depth, so
using the following expression:
that the reaction vials fit; mixing of the contents of the
vial is effected using a magnetic stirrer equipped in the Q x m1 x M1 x 100
heating module or using a reciproca! shaker that performs Q1 x m M2
approximately 100 cycles/min.
Interna[ standard solution. 30 g/L solution of octane R in mass of isopropyl iodide in the reference solution,
o-xylene R. in milligrams;
Test solution. Weigh 65.0 mg of the substance to be examined, m mass of the sample (dried substance), in
place in a reaction vial, add 0.06-0.10 g of adipic acid R, 2.0 mL milligrams;
of the internal standard solution and 2.0 mL of hydriodic
molar mass ofhydroxypropoxy group (75.1);
acid R, immediately cap and seal the vial, and weigh accurately.
Mix the contents of the vial continuously for 60 min while molar mass of isopropyl iodide (170.0).
heating the block so that the temperature of the contents is
maintained at 130 ± 2 ºC. If a reciproca! shaker or magnetic
stirrer cannot be used, shake the vial thoroughly by hand at STORAGE
5 min intervals during the initial 30 min of the heating time.
Allow the vial to cool, and again weigh accurately. If the loss In an airtight container.
of mass is less than 26 mg and there is no evidence of a leak,
use the upper layer of the mixture as the test solution. FUNCTIONALITY-RELATED CHARACTERISTICS
Reference solution. Place 0.06-0.10 g of adipic acid R, 2.0 mL
This section provides information on characteristics that are
of the interna! standard solution and 2.0 mL of hydriodic
recognised as being relevant control parameters for one or
acid R in another reaction vial, cap and seal the vial, and
more functions of the substance when used as an excipient
weigh accurately. Add 15-22 µL of isopropyl iodide R through
(see chapter 5.15). So me of the characteristics described in
the septum with a syringe, and weigh accurately. Shake
the Functionality-related characteristics section may also be
the reaction vial thoroughly and use the upper layer as the
present in the mandatory part of the monograph since they
reference solution.
also represent mandatory quality criteria. In such cases, a
Column: cross-reference to the tests described in the mandatory part is
- material: fused silica; included in the Functionality-related characteristics section.
Control of the characteristics can con tribute to the quality
- size: l = 30 m, 0 = 0.53 mm; of a medicinal product by improving the consistency of the
- stationary phase: poly(dimethyl)siloxane R (3 µm). manufacturing process and the performance of the medicinal
Use a precolumn if needed. product during use. Where control methods are cited, they are
recognised as being suitable f or the purpose, but other methods
Carrier gas: helium for chromatography R. can also be used. Wherever results f ora particular characteristic
Flow rate: 4.3 mL/min. are reported, the control method must be indicated.
Split ratio: 1:40. The following characteristics may be relevant for low-substituted
Temperature: hydroxypropylcellulose used as disintegrant.
Time Temperature Settling volume: 20.0 mL to 35.0 mL.
(min) (ºC) To 20 mL of 2-propanol R in a 100 mL graduated cylinder add
Column o-3 50 5.0 g of the substance to be examined and shake vigorously.
3-8 50 -7 100 Dilute to 30 mL with 2-propanol R, then dilute to 50 mL with
water R and shake vigorously. Within 15 min, repeat the
8 - 12 100 -7 250 shaking 3 times. Seal the cylinder to avoid evaporation of the
12 - 20 250 solvent. Allow to stand for 2 h and determine the volume of
the settled mass.
Injection port 250
Degree of substitution (see Assay).
Detector 280
Partide-size distribution (2.9.31).

EUROPEAN PHARMACOPOEIA 9.5 Hydroxyzine hydrochloride
01/2017:0916 15 min. A precipitate is formed. Filter. Recrystallise from

corrected 9.5 ethanol (96 per cent) R. Initiate crystallisation, if necessary,
by scratching the wall of the tube with a glass rod. The
crystals melt (2.2.14) at 189 ºC to 192 ºC.
D. It gives reaction (a) of chlorides (2.3.1).
HYDROXYZINE HYDROCHLORIDE TESTS

Solution S. Dissolve 2.0 g in water R and dilute to 20.0 mL
Hydroxyzini hydrochloridum with the same solvent.
Appearance of solution. Solution Sis clear (2.2.1) and not
more intensely coloured than reference solution Y7 (2.2.2,
C I D x /I" - N ~O~ OH Method II).
1
~ NJ 2 HCI Optical rotation (2.2.7): - 0.10º to+ 0.10º, determined on
solution S.
O" and enantiomer
Test solution. Dissolve 10.0 mg of the substance to be
C21 H 29 Cl 3 N 2 0 2 examined in the mobile phase and dilute to 10.0 mL with the
[2192-20-3] mobile phase.
DEFINITION Reference solution (a). Dissolve 10.0 mg of hydroxyzine
(RS)-2-[2-[ 4-[ (4-Chlorophenyl)phenylmethyl]piperazin-1- hydrochloride CRS in the mobile phase and dilute to 10.0 mL
yl] ethoxy] ethanol dihydrochloride. with the mobile phase.
Content: 99.0 per cent to 101.0 per cent (dried substance). Reference solution (b ). Dilute 3.0 mL of the test solution to
CHARACTERS to 25.0 mL with the mobile phase.
Appearance: white or almost white, hygroscopic, crystalline Column:
powder. - size: l = 0.15 m, 0 = 4.6 mm;
Solubility: freely soluble in water and in ethanol (96 per cent), - stationary phase: base-deactivated end-capped octadecylsilyl
very slightly soluble in acetone. silica gel for chromatography R (3 µm).
mp: about 200 ºC, with decomposition. Mobile phase: dissolve 0.5 g of sodium methanesulfonate R in a
IDENTIFICATION mixture of 14 mL of triethylamine R, 300 mL of acetonitrile for
chromatography R and 686 mL of water far chromatography R,
First identification: A, D. then adjust to pH 2.7 with sulfuric acid R.
Second identification: B, C, D. Flow rate: 1 mL/min.
-
Comparison: hydroxyzine hydrochloride CRS.

B. Thin-layer chromatography (2.2.27).
Injection: 20 µL.
Run time: 2.5 times the retention time of hydroxyzine.
Solvent mixture: methanol R, methylene chloride R System suitability: reference solution (a):
(50:50 V/V). - peak-to-valley ratio: minimum 10, where HP = height above
Test solution. Dissolve 0.50 g of the substance to be the baseline of the peak immediately before the peak due
examined in the solvent mixture and dilute to 10 mL with to hydroxyzine and Hv = height above the baseline of the
the solvent mixture. lowest point of the curve separating this peak from the
Reference solution (a). Dissolve 0.50 g of hydroxyzine peak due to hydroxyzine.
hydrochloride CRS in the solvent mixture and dilute to Limits:
1O mL with the solvent mixture. - any impurity: for each impurity, not more than 1/3 of the
Reference solution (b ). Dissolve O.SO g of meclozine area of the principal peak in the chromatogram obtained
dihydrochloride R in the solvent mixture and dilute to with reference solution (b) (O.l per cent);
1O mL with the solvent mixture. Dilute 1 mL of the solution - total: not more than the area of the principal peak in the
to 2 mL with reference solution (a). chromatogram obtained with reference solution (b) (0.3 per
Plate: TIC silica gel G plate R. cent);
Mobile phase: concentrated ammonia R, ethanol (96 per - disregard limit: 0.1 times the area of the principal peak in
cent) R, toluene R (1:24:75 V/V/V). the chromatogram obtained with reference solution (b)
Application: 2 µL. (0.03 per cent).
Development: overa path of 15 cm. Loss on drying (2.2.32): maximum 5.0 per cent, determined
Drying: in air.
Detection: spray with potassium iodobismuthate solution R2. Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
1.0 g.
- the chromatogram shows 2 clearly separated principal ASSAY
spots. Dissolve 0.200 g in 10 mL of anhydrous acetic acid R. Add
Results: the principal spot in the chromatogram obtained 40 mL of acetic anhydride R. Titrate with 0.1 M perchloric
with the test solution is similar in position, colour and size acid, determining the end-point potentiometrically (2.2.20).
to the principal spot in the chromatogram obtained with 1 mL of 0.1 M perchloric acid is equivalent to 22.39 mg
reference solution (a). of C 21 H 29 Cl 3 N 2 0 2 •
C. Dissolve 0.1 gin ethanol (96 per cent) R and dilute to 15 mL
with the same solvent. Add 15 mL of a saturated solution of STORAGE
picric acid R in ethanol (96 per cent) R. Allow to stand for In an airtight container, protected from light.
Hyoscine butylbromide EUROPEAN PHARMACOPOEIA 9.5
-
IMPURITIES Appearance of solution. Solution S is clear (2.2.1) and
CIDxOH colourless (2.2.2, Method JI).
Acidity or alkalinity. Dissolve 1.0 g in carbon dioxide-free
OH and enantiomer
water R and dilute to 1O mL with the same solvent. Add
0.25 mL of 0.01 M sodium hydroxide and 0.2 mL of methyl red
mixed solution R. The solution is green. Not more than 0.5 mL
of 0.01 M hydrochloric acid is required to change the colour of
A. (RS)-l-[ (4-chlorophenyl)phenylmethyl]piperazine, the indicator to reddish-violet.
rN~O~OH Specific optical rotation (2.2.7): - 20 to - 18 (dried

N-J Related substances. Liquid chromatography (2.2.29).
1 examined in mobile phase B and dilute to 10.0 mL with mobile
":::::,...
phase B.
B. 2-[2-[ 4-( diphenylmethyl)piperazin-1-yl]ethoxy]ethanol Reference solution (a). Dilute 1.0 mL of the test solution to
(decloxizine ). 100.0 mL with mobile phase B. Dilute 1.0 mL of this solution
to 10.0 mL with mobile phase B.
Reference solution (b). Dissolve 5 mg of hyoscine butylbromide
07/2018:0737 for system suitability CRS (containing impurities A and B) in
mobile phase B and dilute to 10.0 mL with mobile phase B.
Column:
- size: l = 0.10 m, 0 = 4.6 mm;
HYOSCINE BUTYLBROMIDE - stationary phase: silica gel for chromatography, alkyl-bonded
for use with highly aqueous mobile phases R (l.8 µm);
Hyoscini butylbromidum - temperature: 50 ºC.
Mobile phase:
Scopolamini butylbromidum - mobile phase A: acetonitrile Rl, 0.2 per cent V/V solution
of perchloric acid R (5:95 V/V);
- mobile phase B: 0.2 per cent Vl\1 solution of perchloric
acid R, acetonitrile Rl (30:70 V/V);
o- 1 91 9
1 - 4.2 91 -7 75 9 -7 25
C21 H 30 BrN0 4
[149-64-4] 4.2 - 5.5 75 -7 66 25 -7 34
5.5 - 10 66 -7 15 34 -7 85
DEFINITION
( lR,2R,4S,5S,7s,9r)-9-Butyl-7- [[(2S)-3-hydroxy-2-phenyl- 10 - 11 15 85
propanoyl]oxy ]-9-methyl-3-oxa-9-azatricyclo[3.3. l .02.4]non-
an-9-ium bromide. Flow rate: 2.5 mL/min.
Content: 98.0 per cent to 101.0 per cent (dried substance). Detection : spectrophotometer at 21 O nm.
Injection: 2 µL.
CHARACTERS
Appearance: white or almost white, crystalline powder. with hyoscine butylbromide for system suitability CRS and the
Solubility: freely soluble in water and in methylene chloride, chromatogram obtained with reference solution (b) to identify
sparingly soluble in anhydrous ethanol. the peaks due to impurities A and B.
IDENTIFICATION Relative retention with reference to hyoscine butylbromide
(retention time= about 4.8 min): bromide = about 0.1;
First identification: A, C, F. impurity B = about 0.32; impurity A= about 0.37.
Second identification: A, B, D, E, F. System suitability: reference solution (b):
A. Specific optical rotation (see Tests). - resolution: mínimum 2.0 between the peaks due to
B. Melting point (2.2.14): 139 ºC to 141 ºC. impurities B and A.
C. Infrared absorption spectrophotometry (2.2.24). Calculation of percentage contents:
Comparison: hyoscine butylbromide CRS. - for each impurity, use the concentration of hyoscine
D. To about 1 mg add 0.2 mL of nitric acid R and evaporate to butylbromide in reference solution (a).
dryness on a water-bath. Dissolve the residue in 2 mL of Limits:
acetone R and add 0.1 mL of a 30 g/L solution of potassium - impurity A: maximum 0.15 per cent;
hydroxide R in methanol R. A violet colour develops.
E. To 5 mL of solution S (see Tests) add 2 mL of dilute sodium 0.10 per cent;
hydroxide solution R. No precipitate is formed.
F. It gives reaction (a) of bromides (2.3.1).
- reporting threshold: 0.05 per cent; disregard the peak due
TESTS to bromide.
Solution S. Dissolve 1.25 g in water R and dilute to 25.0 mL Loss on drying (2.2.32): maximum 2.5 per cent, determined
with the same solvent. on 0.500 g by drying in an oven at 105 ºC.

EUROPEAN PHARMACOPOEIA 9.5 Hyoscine butylbromide

0.5 g.
ASSAY
Dissolve 0.400 g in 50 mL of water R. Titrate with 0.1 M silver
nitrate, determining the end-point potentiometrically (2.2.20)
using a silver indicator electrode and a silver-silver chloride D. (1R,2R,4S,5S,7s,9r)-7-[[(2S)-3-hydroxy-2-phenyl-
reference electrode. propanoyl] oxy]-9-methyl-9-propyl-3-oxa-9-azatricyclo-
[3.3. l.02.4] nonan-9-ium (propylhyoscine),
1 mL of 0.1 M silver nitrate is equivalent to 44.04 mg
of C21 H 30BrN0 4 •
IMPURITIES
Specified impurities: A.
the tests in the monograph. They are limited by the general E. ( 1R,2R,4S,5S,7s )-9-butyl-3-oxa-9-azatricyclo [3.3.1.0 2·4]-
acceptance criterion for other/unspecified impurities and/or nonan-7 -yl ( 2S)-3-hydroxy-2-phenylpropanoate
by the general monograph Substances for pharmaceutical (N-butylnorhyoscine),
impurities for demonstration of compliance. See also 5.1 O.
Control of impurities in substances for pharmaceutical use): B,
H )H~
óxl>C~::hº
C, D, E, F, G, H.
~
\H
Q/>(-Yº
~ \ H
OH
F. ( IR,2R,4S,5S,7 s,9s )-9-butyl-7-[ [(2S)-3-hydroxy-

2-phenylpropanoyl] oxy]-9-methyl-3-oxa-9-
OH H H
azatricyclo[3.3. l .0 2.4]nonan-9-ium,
H~---H~~CH3
A. ( IR,2R,4S,5S,7 s)-9-methyl-3-oxa-9-azatricyclo [3.3.1.02.4]-
nonan-7 -yl ( 2S)-3-hydroxy-2-phenylpropanoate (hyoscine), /H
º. . ---~l- \ o
°<:co,H and eoam;ome•
CH2 H l .H
CH3
OH G. ( 1R,2R,4S,5S, 7s,9r)-9-butyl-9-methyl-7-[ (2-phenylprop-

2-enoyl)oxy]-3-oxa-9-azatricyclo [3.3.1.02.4] nonan-9-ium
B. (2RS)-3-hydroxy-2-phenylpropanoic acid (DL-tropic acid), (apo-N-butylhyoscine),
H
o °<{:~-~
OH H l
C. ( 1R,2R,4S,5S,7s )-7-[ [(2S)-3-hydroxy-2-phenylpropanoyl]- H. (1R,3r,5S,8s)-8-butyl-3-[[(2S)-3-hydroxy-2-phenyl-
oxy]-9,9-dimethyl-3-oxa-9-azatricyclo[3.3. l .02'4]nonan-9- propanoyl]oxy]-8-methyl-8-azabicyclo[3.2.l]octan-8-ium
ium (methylhyoscine), (N-butylhyoscyamine).

1
Insulin glargine ....................................................................... 5699 Isoniazid................................................................................... 5703
Interferon beta-la concentrated solution ............................ 5701 Isotretinoin .............................................................................. 5705

EUROPEAN PHARMACOPOEIA 9.5 Insulin glargine
07 /2018:2571 and 100 µL of a 20 U/mL solution of Staphylococcus aureus

strain V8 protease, type XVII-B R in 1 M tris-hydrochloride
buffer solution pH 7.5 R. Mix and incubate at 45 ºC for
about 2 h. Stop the reaction by adding 2 µL of phosphoric
acid R.
INSULIN GLARGINE Reference solution. Prepare at the same time and in the
same manner as for the test solution but using insulin
glargine CRS instead of the substance to be examined.
Insulinum glarginum
CHROMA TOGRAPHIC SEPARA TION. Liquid chromatography
(2.2.29).
H - Gly - lle - Val - Glu - Gin - Cys - Cys - Thr - Ser - lle -
JO Buffer solution. Dissolve 11.6 g of phosphoric acid R and
- ~ys
Cys - Ser - Leu -Tyr - Gin - Leu - Glu - Asn - T y ; J- r
42.1 g of sodium perchlorate R in 1600 mL of water for
chromatography R, adjust to pH 2.3 with triethylamine R
Gly-OH and dilute to 2000 mL with water for chromatography R.
Column:
- size: l = 0.125 m, 0 = 3.0 mm;
- stationary phase: spherical end-capped octadecylsilyl
Glu - Arg- Gly - Phe - Phe -Tyr -Thr - Pro - Lys - ~~r - silica gel for chromatography R (4 µm);
Arg-Arg-OH
Mobile phase:
Mr6063
- mobile phase A: acetonitrile Rl, buffer solution
DEFINITION (7:93 V/V);
21 A-Glycine-30 8 a-L-arginine-30 8 b-L-arginine-insulin - mobile phase B: buffer solution, acetonitrile Rl
(human). (43:57 V!V);
2-chain peptide containing 53 amino acids. The A-chain is Time Mobile phase A Mobile phase B
composed of 21 amino acids and the B-chain is composed of (min) (per cent V/V) (per cent V/V)
32 amino acids. It is identical in primary structure to human o - 30 90 7 20 10 7 80
insulin, only differing in amino acid sequence at position 21
30 - 35 20 80
in the A-chain and at the C-terminal end of the B-chain
where it contains 2 additional amino acids. Human insulin is
Asn(A21), whereas insulin glargine is Gly(A21), Arg(B31),
Arg(B32). As in human insulin, insulin glargine contains Detection: spectrophotometer at 214 nm.
2 interchain disulfide bonds and 1 intrachain disulfide bond. Equilibration: at initial conditions for at least 15 min.
Content: 94.0 per cent to 105.0 per cent (anhydrous substance). Injection: 50 µL.
By convention, for the purpose of labelling insulin glargine Retention time: insulin glargine fragment II = about
preparations, 0.0364 mg of insulin glargine is equivaleñt to 14 min; insulin glargine fragment III = about 15 min.
1 unit. System suitability:
PRODUCTION - the chromatograms obtained with the test solution and
the reference solution are qualitatively similar to the
Insulin glargine is produced by a method based on chromatogram of insulin glargine digest supplied with
recombinant DNA (rDNA) technology under conditions insulin glargine CRS;
designed to minimise the degree of microbial contamination.
- in the chromatogram obtained with the reference
Prior to releas e, the f ollowing tests are carried out on each batch solution, identify the peaks due to digest fragments II
of the final bulk product, unless exemption has been granted by and III:
the competent authority.
symmetry factor: maximum 1.5 for the peaks dueto
Host-cell-derived proteins. The limit is approved by the fragments II and III;
resolution: minimum 3.4 between the peaks due to
Single-chain precursor. The limit is approved by the fragments II and III.
competent authority. Use a suitably sensitive method. Results: the profile of the chromatogram obtained with the test
CHARACTERS solution corresponds to that of the chromatogram obtained
with the reference solution.
Appearance: white or almost white, hygroscopic powder.
NOTE: the retention times offragments I and IV are the same
Solubility: practically insoluble in water and in anhydrous as for human insulin; the retention times offragments JI and III
ethanol, soluble in dilute mineral acids. differ from human insulin due to the difference in the sequence
IDENTIFICATION at position 21 of the A-chain and to the 2 additional amino
acids of the B-chain.
A. Examine the chromatograms obtained in the assay.
Results: the principal peak in the chromatogram obtained TESTS
with the test solution is similar in retention time to the Impurities with molecular masses greater than that of
principal peak in the chromatogram obtained with the insulin glargine. Size-exclusion chromatography (2.2.30): use
reference solution. the normalisation procedure.
B. Peptide mapping (2.2.55). Test solution. Prepare a 4 mg/mL solution of the substance to
SELECTIVE CLEAVAGE OF THE PEPTIDE BONDS be examined in a 1 g/L solution of hydrochloric acid R.
Test solution. Prepare a 10.0 mg/mL solution of the Resolution solution. Dry about 200 mg of the substance
substance to be examined in a 1 g/L solution of hydrochloric to be examined in an oven at 100 ºC for 1.5-3 h. Dissolve
acid R and transfer 5 µL of the solution to a clean tube. Add 15.0 mg of the dried substance in 1.5 mL of a 1 g/L solution of
1.0 mL of 1 M tris-hydrochloride buffer solution pH 7.5 R hydrochloric acid R and dilute to 10.0 mL with water R.
Insulin glargine EUROPEAN PHARMACOPOEIA 9.5
Reference solution. Dilute 1.0 mL of the test solution to Mobile phase:

100.0 mL with water R. Dilute 3.0 mL of this solution to - mobile phase A: dissolve 18.4 g of sodium chloride R
20.0 mL with water R. in 250 mL of the buffer solution, add 250 mL of
Column: acetonitrile Rl and mix; dilute to 1000 mL with water for
chromatography R;
- size: l = 0.3 m, 0 = 7.8 mm; mobile phase B: dissolve 3.2 g of sodium chloride R in 250 mL
- stationary phase: hydrophilic silica gel for chromatography R of the buffer solution, add 650 mL of acetonitrile Rl and
(10 µm) with a pore size of 12.5 nm, of a grade suitable for mix; dilute to 1000 mL with water for chromatography R;
fractionation of globular proteins in the relative molecular Time Mobile phase A Mobile phase B
mass range of 5000 to 150 000. (min) (per cent V/V) (per cent V/V)
Mobile phase: mix 15 volumes of glacial acetic acid R, o - 20 96 -7 83 4 -7 17
20 volumes of acetonitrile R and 65 volumes of a 1.0 g/L 20 - 30 83 -7 63 17 -7 37
solution of arginine R; fil ter and <legas.
30 - 33 63 -7 96 37 -7 4
33 - 40 96 4
Injection: 100 µL.
Run time: about 35 min. Injection: 5 µL of the test solution and the resolution solution.
Retention time: insulin glargine = about 18 min. Retention time: insulin glargine = about 20 min.
System suitability: System suitability: resolution solution:

- peak-to-valley ratio: mínimum 2, where HP = height above
- signal-to-noise ratio: mínimum 10 for the principal peak in the baseline of the peak dueto OA-Arg-insulin glargine and
the chromatogram obtained with the reference solution; Hv = height above the baseline of the lowest point of the
- symmetry factor: maximum 2.0 for the peak dueto insulin
curve separating this peak from the peak due to insulin
glargine in the chromatogram obtained with the resolution glargine.
solution; Limits:
- peak-to-valiey ratio: mínimum 2.0, where HP = height - any impurity: for each impurity, maximum 0.4 per cent;
above the baseline of the peak due to high molecular mass - total: maximum 1.0 per cent.
proteins and Hv = height above the baseline of the lowest
Zinc: maximum 0.80 per cent.
point of the curve separating this peak from the peak due
to insulin glargine in the chromatogram obtained with the Atomic absorption spectrometry (2.2.23, Method I).
resolution solution. Test solution. Dissolve 45.0 mg of the substance to be
Limits: examined in a 1 g/L solution of hydrochloric acid R and dilute
to SO.O mL with the same solution. Dilute 10.0 mL of the
- total of impurities with a retention time less than that of solution to 100.0 mL with a 1 g/L solution of hydrochloric
insulin glargine: maximum 0.3 per cent of the total area of acid R.
the peaks; disregard any peak with a retention time greater Reference solutions. Prepare reference solutions containing
than that of the peak due to insulin glargine. 0.2 µg, 0.4 µg and 0.6 µg of zinc per millilitre by diluting
Related proteins. Liquid chromatography (2.2.29): use the zinc standard solution (10 ppm Zn) R with a 1 g/L solution
normalisation procedure. Maintain the solutions at 2-8 ºC. of hydrochloric acid R.
Source: zinc hollow-cathode lamp.
examined in 1.5 mL of a 1 g/L solution of hydrochloric acid R Wavelength: 213.9 nm.
and dilute to 10.0 mL with water R. Atomisation device: air-acetylene flame of suitable composition
Reference solution. Dissolve the contents of a vial of insulin (for example, 11 L of air and 2 L of acetylene per minute).
glargine CRS in 1.5 mL of a 1 g/L solution of hydrochloric Water (2.5.32): maximum 8.0 per cent, determined on
acid R, transfer the solution with water R to a 1O mL 30.0 mg.
volumetric flask and dilute to 10.0 mL with water R. Bacteria! endotoxins (2.6.14, Method D): less than 10 IU/mg,
Resolution solution. Dissolve the contents of a vial of insulin if intended for use in the manufacture of parenteral
glargine for peak identification CRS (containing OA-Arg-insulin preparations without a further appropriate procedure for the
glargine) in 0.3 mL of a 1 g/L solution of hydrochloric acid R removal of bacteria! endotoxins.
and add 1. 7 mL of water R.
ASSAY
Buffer solution. Dissolve 20.7 g of anhydrous sodium
dihydrogen phosphate R in 900 mL of water for
related proteins with the following modification.
chromatography R, adjust to pH 2.5 with phosphoric acid R
and dilute to 1000 mL with water for chromatography R. Injection: 5 µL of the test solution and the reference solution.
Column: Calculate the content of insulin glargine (C 267 H 404N 72 Ü 78 S6 )

taking into account the assigned content of insulin
- size: l = 0.25 m, 0 = 3.0 mm; glargine CRS.
- stationary phase: spherical end-capped octadecylsilyl silica STORAGE

gel for chromatography R (4 µm);
- temperature: 35 ºC. of - 20 ± 5 ºC.

EUROPEAN PHARMACOPOEIA 9.5 Interferon beta- la concentrated solution
01/2009:1639 Interpretation of results: a typical spectrum consists

corrected 9.5 of 6 major glycoforms (A to F), which differ in their
degree of sialylation and/or antennarity type as shown in
Table 1639.-1.
Table 1639.-1.
MS peak Glycoform* Expected M, Sialylation
INTERFERON BETA-la level
CONCENTRATED SOLUTION A 2A2S1F 22 375 Disialylated
B 2A1S1F 22 084 Monosialylated
Interferoni beta-la solutio concentrata e 3A2S1F and/or 22 739 Disialylated
2A2S1F
MSYNLLGFLQ RSSNFQCQKL LWQLNGRLEY CLKDRMNFDI + 1 HexNacHex
PEEIKQLQQF QKEDAALTIY EMLQNIFAIF RQDSSSTGWN repeat
ETIVENLLAN VYHQINHLKT VLEEKLEKED FTRGKLMSSL D 3A3S1F 23 031 Trisialylated
HLKRYYGRIL HYLKAKEYSH CAWTIVRVEI LRNFYFINRL
E 4A3S1F and/or 23 400 Trisialylated
TGYLRN
3A3S1F
* glycosylation site + 1 HexNacHex
repeat
C9osH14o6N24602s2s7 Mr approx. 22 500
F 2AOS1F 21 793 Non-sialylated
DEFINITION * 2A = biantennary complex type oligosaccharide; 3A = triantennary
Solution of a glycosylated protein having the same amino acid complex type oligosaccharide; 4A = tetraantennary complex
type oligosaccharide; OS = non-sialylated; 1S = monosialylated;
sequence and disulfide bridge and a similar glycosylation 2S = disialylated; 3S = trisialylated; lF = fucosylated.
pattern as interferon beta produced by human diploid
fibroblasts in response to viral infections and various Results: the mass spectrum obtained with the preparation
other inducers. It exerts antiviral, antiproliferative and to be examined corresponds, with respect to the 6 major
immunomodulatory activity. peaks, to the mass spectrum obtained with interferon
beta-la CRS.
Content: minimum 0.20 mg of protein per millilitre.
C. Peptide mapping (2.2.55) and liquid chromatography
Potency: minimum 1.5 x 10 8 IU per milligram of protein. (2.2.29).
It may contain buffer salts. Test solution. Add 5 µL of a 242 g/L solution of
tris(hydroxymethyl)aminomethane R and a volume of the
PRODUCTION preparation to be examined containing 20 µg of protein
Interferon beta-la concentrated solution is produced by a to a polypropylene tube of 0.5 mL capacity. Add 4 µL of
method based on recombinant DNA (rDNA) technology, a 1 mg/mL solution of endoprotease LysC R in 0.05 M
using mammalian cells in culture. tris-hydrochloride buffer solution pH 9.0 R. Mix gently and
Prior to release, the following tests are carried out on each incubate at 30 ºC for 2 h. Add 10 µL of a 15.4 g/L solution
batch of the final bulk product, unless exemption has been of dithiothreitol R. Dilute the solution with the same
granted by the competent authority. volume of a 573 g/L solution of guanidine hydrochloride R.
Incubate at 4 ºC for 3-4 h.
Host-cell-derived proteins. The limit is approved by the
Reference solution. Prepare at the same time and in the
same manner as for the test solution but using interferon
Host-cell or vector-derived DNA. The limit is approved by beta-1 a CRS instead of the preparation to be examined.
the competent authority. Precolumn:
N-terminal truncated forms. Examination for specific - size: l = 0.02 m, 0 = 2.1 mm;
N-terminal truncated forms should be performed using a - stationary phase: spherical octadecylsilyl silica gel for
suitable technique such as N-terminal sequence determination. chromatography R (5 µm) with a pore size of 30 nm.
The limits are approved by the competent authority.
Column:
Dimer and related substances of higher molecular mass: not - size: l = 0.25 m, 0 = 2.1 mm;
more than the amount approved by the competent authority,
using an appropriate validated liquid chromatography method. - stationary phase: spherical octadecylsilyl silica gel for
chromatography R (5 µm) with a pore size of 30 nm.
CHARACTERS Mobile phase:
Appearance: clear or slightly opalescent, colourless or slightly - mobile phase A : dilute 1 mL of trifluoroacetic acid R to
yellowish liquid. 1000 mL with water for chromatography R;
- mobile phase B: dilute 1 mL of trifluoroacetic acid R in
IDENTIFICATION 700 mL of acetonitrile Rl, then dilute to 1000 mL with
A. It shows the expected biological activity (see Assay). water for chromatography R;
B. Isoform distribution. Mass spectrometry (2.2.43). Time Mobile phase A Mobile phase B
Introduction of the sample: direct inflow of a desalted
preparation to be examined or liquid chromatography-mass o - 30 100-? 64 o 7 36
spectrometry combination. 30 - 45 64 7 55 36 7 45
Mode of ionisation: electrospray. 45 - 50 55-? 40 45 7 60
Signal acquisition: complete spectrum mode from 50 - 70 40 7 o 60 7 100
1100 to 2400.
Calibration: use myoglobin in the miz range of 600-2400; 70 - 83 o 100
set the instrument within validated instrumental settings 83 - 85 o7 100 100 7 o
and analyse the sample; the deviation of the measured mass
does not exceed 0.02 per cent of the reported mass. Flow rate: 0.2 mL/min.
Interferon beta-la concentrated solution EUROPEAN PHARMACOPOEIA 9.5
Detection: spectrophotometer at 214 nm. - in the electropherogram obtained with test solution (b ),
Injection: volume that contains 20 µg of digested protein. the band corresponding to deglycosylated interferon
beta-1 a is not more intense than the principal band in the
System suitability: the chromatogram obtained with electropherogram obtained with test solution (e) (2 per
the reference solution is qualitatively similar to the cent); any other band corresponding to an impurity of
chromatogram of interferon beta-la digest supplied with a molecular mass lower than that of interferon beta-1 a,
interferon beta-la CRS. apart from the band corresponding to underglycosylated
Results: the profile of the chromatogram obtained with interferon beta-1 a is not more intense than the principal
the test solution corresponds to that of the chromatogram band in the electropherogram obtained with test solution (f)
obtained with the reference solution. (1 per cent).
TESTS Oxidised interferon beta- la: maximum 6 per cent.
Impurities of molecular masses differing from that of Use the chromatogram obtained with the test solution in
interferon beta-la. Polyacrylamide gel electrophoresis identification C. Locate the peaks due to the peptide fragment
(2.2.31) under reducing conditions. comprising amino acids 34-45 and its oxidised form using the
chromatogram of oxidised interferon beta-la digest supplied
Resolving gel: 12 per cent acrylamide.
with interferon beta-1 a CRS.
Concentrated sample buffer: concentrated SDS-PAGE sample
Calculate the percentage of oxidation of interferon beta-la
buffer for reducing conditions R containing 2-mercaptoethanol
as the reducing agent. using the following expression:
Sample buffer: mixture of equal volumes of concentrated A34-45ox X lQO
SDS-PAGE sample buffer for reducing conditions R and water R. A34-45 + A34-45ox
Test solution (a). Concentrate the preparation to be examined
using a suitable method to obtain a protein concentration of A34-45ox area of the peak due to the oxidised peptide
1.5 mg/mL. fragment 34-45;
Test solution (b): mixture of equal volumes of test solution (a) area of the peak due to the peptide fragment
and the concentrated sample buffer. 34-45.
Test solution (e). Dilute test solution (a) to obtain a protein Bacteria! endotoxins (2.6.14): less than 0.7 IU in the volume
concentration of 0.6 mg/mL. Mix equal volumes of this that contains 1 x 106 IU of interferon beta-1 a, if intended for
solution and the concentrated sample buffer. use in the manufacture of parenteral preparations without
Test solution (d). Mix 8 µL of test solution (c) and 40 µL of a further appropriate procedure for removal of bacteria!
the sample buffer. endotoxins.
Test solution (e). Mix 15 µL of test solution (d) and 35 µL of
the sample buffer. ASSAY
Test solution (f). Mix 18 µL of test solution (e) and 18 µL of Protein. Liquid chromatography (2.2.29). Prepare
the sample buffer. 3 independent dilutions for each solution.
Test solution (g). Mix 12 µL of test solution (f) and 12 µL of Test solution. Dilute the preparation to be examined to obtain
the sample buffer. a concentration of 100 µg/mL.
Reference solution. Solution of relative molecular mass Reference solution. Dissolve the contents of a vial of interferon
markers suitable for calibrating SDS-PAGE gels in the range of beta-la CRS to obtain a concentration of 100 µg/mL.
15-67 kDa. Dissolve in the sample buffer. Precolumn:
Sample treatment: boil for 3 min. - size: l = 0.02 m, 0 = 2.1 mm;
Application: 20 µL of test solutions (b) to (g) and the reference _ stationary phase: end-capped butylsilyl silica gel for
solution. chromatography R (5 µm) with a pore size of 30 nm.
Detection: Coomassie staining, carried out as follows: Column:
immerse the gel in Coomassie staining solution Rl at 33-37 ºC
for 90 min with gentle shaking, then remove the staining - size: l = 0.25 m, 0 = 2.1 mm;
solution; destain the gel with a large excess of a mixture of - stationary phase: end-capped butylsilyl silica gel for
1 volume of glacial acetic acid R, 1 volume of 2-propanol R and chromatography R (5 µm) with a pore size of 30 nm.
8 volumes of water R. Mobile phase:
Apparent molecular masses: interferon beta-la= about 23 000; - mobile phase A: 0.1 per cent V/V solution of trifluoroacetic
underglycosylated interferon beta-la= about 21 000; acid R;
deglycosylated interferon beta-la= about 20 000; interferon
beta-la dimer = about 46 000. - mobile phase B: to 300 mL of water for chromatography R,
add 1 mL of trifluoroacetic acid R and dilute to 1000 mL
Identification of bands: use the electropherogram provided with acetonitrile Rl;
with ínterferon beta-la CRS.
System suitabílity: Time Mobile phase A Mobile phase B
the validation criteria are met (2.2.31);
o - 20 100 -7 o o -7 100
- a band is seen in the electropherogram obtained with test
solution (g) ; 20 - 25 o 100
- a gradation of intensity of staining is seen in the 25 - 26 o -7 100 100 -7 o

electropherograms obtained with test solutions (b) to (g).
26 - 40 100 o
Limits:
- in the electropherogram obtained with test solution (c), Flow rate: 0.2 mL/min.
the band corresponding to underglycosylated interferon Detection: spectrophotometer at 214 nm.
beta-la is not more intense than the principal band in the
electropherogram obtained with test solution (e) (5 per Injection: 50 µL.
cent); Retention time: interferon beta-la= about 20 min.

EUROPEAN PHARMACOPOEIA 9.S Isoniazid
System suitability: reference solution: 07/2018:0146

- symmetry factor: 0.8 to 2.0 for the peak dueto interferon
beta-la;
3.0 per cent between the peak areas obtained after injection ISONIAZID
of the 3 independent dilutions.
Calculate the content of interferon beta-la (C908 H 1406N 246 Ü 252 S7 )
Isoniazidum
taking into account the assigned content of C908 H 1406N 246 Ü 252 S7
in interferon beta-la CRS.
Potency
The potency of interferon beta-la is estimated by comparing
its ability to protect cells against a viral cytopathic effect with C6H 7N 30
the same ability of the appropriate International Standard [S4-8S-3]
of human recombinant interferon beta-la or of a reference
preparation calibrated in International Units. DEFINITION
Pyridine-4-carbohydrazide.
The International Unit is the activity contained in a stated Content: 99.0 per cent to 101.0 per cent (dried substance).
amount of the appropriate International Standard. The
equivalence in International Units of the International CHARACTERS
Standard is stated by the World Health Organization. Appearance: white or almost white, crystalline powder or
Carry out the assay using a suitable method, based on the colourless crystals.
following design. Solubility: freely soluble in water, sparingly soluble in ethanol
(96 per cent).
Use, in standard culture conditions, an established cell line
sensitive to the cytopathic effect of a suitable virus and IDENTIFICATION
responsive to interferon. The cell cultures and viruses that First identification: A, B.
have been shown to be suitable include the following: Second identification: A, C.
- WISH cells (ATCC No. CCL-25) and vesicular stomatitis A. Melting point (2.2.14) : 170 ºC to 174 ºC.
virus VSV, Indiana strain (ATCC No. VR-1S8) as infective B. Infrared absorption spectrophotometry (2.2.24).
agent; Comparison: isoniazid CRS.
- A549 cells (ATCC No. CCL-18S) and encephalomyocarditis C. Dissolve 0.1gin2 mL of water R and add 10 mL of a warm
virus EMC (ATCC No. VR-129B) as infective agent. 10 g/L solution of vanillin R. Allow to stand and scratch the
wall of the test tube with a glass rod. A yellow precipitate is
Incubate in at least 4 series, cells with 3 or more different formed, which, after recrystallisation from S mL of ethanol
concentrations of the preparation to be examined and the (70 per cent \'IV) R and drying at 100-105 ºC, melts (2.2.14)
reference preparation in a microtitre plate and include in at 226 ºC to 231 ºC.
each series appropriate controls of untreated cells. Choose
the concentrations of the preparations such that the lowest TESTS
concentration produces sorne protection and the largest Solution S. Dissolve 2.5 g in carbon dioxide-free water R and
concentration produces less than maximal protection dilute to SO mL with the same solvent.
against the viral cytopathic effect. Add at a suitable time the
Appearance of solution. Solution S is clear (2.2.1) and not
cytopathic virus to all wells with the exception of a sufficient
more intensely coloured than reference solution BY 7 (2.2.2,
number of wells in all series, which are left with uninfected
Method JI).
control cells. Determine the cytopathic effect of the virus
quantitatively with a suitable method. Calculate the potency pH (2.2.3): 6.0 to 8.0 for solution S.
of the preparation to be examined by the usual statistical Impurity E. Liquid chromatography (2.2.29). Use freshly
methods (for example, 5.3). prepared solutions.
The estimated potency is not less than 80 per cent and not Solvent mixture: acetonitrile R, water R (SO:SO V/V).
more than 12S per cent of the stated potency. The confidence Solution A. To 2.0 mL of benzaldehyde R add acetonitrile R
limits (P = 0.9S) are not less than 64 per cent and not more and dilute to 100.0 mL with the same solvent. Use the solution
than 1S6 per cent of the estimated potency. within 4 h.
Test solution. Dissolve SO.O mg of the substance to be
examined in 1.0 mL of water R, add S.O mL of solution A and
STORAGE shake well. Wait for 4S min for completion of derivatisation
In an airtight container, protected from light, ata temperature and dilute to 1O.O mL with the solvent mixture.
below - 70 ºC. If the substance is sterile, store in a sterile, Reference solution. Dissolve 20.0 mg of hydrazine sulfate R
airtight, tamper-proof container. (equivalent to 4.92S mg of impurity E) in water R and dilute to
SO.O mL with the same solvent. Dilute 2.5 mL of the solution
to 100.0 mL with water R. To 1.0 mL of this solution add
LABELLING 2.S mL of solution A and shake well. Wait for 4S min for
completion of derivatisation and dilute to 2S.O mL with the
The label states: solvent mixture. Dilute 7.S mL of this solution to 10.0 mL
- the interferon beta-la content, in milligrams per millilitre; with the solvent mixture.
Column:
- the antiviral activity, in International Units per millilitre; - size: l = 0.2S m, 0 = 4.6 mm;
- where applicable, that the substance is suitable for use in - stationary phase: end-capped octadecylsilyl silica gel for
the manufacture of parenteral preparations. chromatography R (S µm).
General Notices (1) apply to ali monographs and other texts S703
Isoniazid EUROPEAN PHARMACOPOEIA 9.S
Mobile phase: water for chromatography R, acetonitrile R Calculation of percentage contents:

(40:60 V/V). - correction factors: multiply the peak areas of the following
Flow rate: 1.0 mL/min. impurities by the corresponding correction factor:
Detection: spectrophotometer at 300 nm. impurity A = 1.4; impurity B = l.S;
Injection: 10 µL. - for each impurity, use the concentration of isoniazid in
Run time: l.S times the retention time of benzaldehyde azine.
Limits:
Retention time: benzaldehyde azine = about 13 min.
- impurities A, B: for each impurity, maximum O.lS per cent;
- repeatability: maximum relative standard deviation of 0.10 per cent;
S.O per cent determined on 6 injections.
- total: maximum O.S per cent;
Calculation of percentage content:
- for impurity E, use the concentration of hydrazine sulfate
in the reference solution. Loss on drying (2.2.32): maximum 0.5 per cent, determined
on 1.000 g by drying in an oven at lOS ºC.
Limit:
- impurity E: maximum lS ppm. 1.0 g.
Related substances. Liquid chromatography (2.2.29). Use
freshly prepared solutions. ASSAY
Test solution. Dissolve 2S.O mg of the substance to be Dissolve 0.2SO g in water R and dilute to 100.0 mL with the
examined in mobile phase A and dilute to 2S.O mL with same solvent. To 20.0 mL of the solution add 100 mL of
mobile phase A. water R, 20 mL of hydrochloric acid R, 0.2 g of potassium
Reference solution (a). Dilute 1.0 mL of the test solution to bromide R and O.OS mL of methyl red solution R. Titrate
100.0 mL with mobile phase A. Dilute 1.0 mL of this solution dropwise with 0.0167 M potassium bromate, shaking
to 1O.O mL with mobile phase A. continuously, until the red colour disappears.
Reference solution (b ). Dissolve S mg of isonicotinic acid R 1 mL of 0.0167 M potassium bromate is equivalent to 3.429 mg
(impurity A), S mg of isonicotinamide R (impurity B) and ofC 6H 7Np.
S mg of nicotinoyl hydrazide R (impurity D) in mobile phase A
IMPURITIES
and dilute to SO.O mL with mobile phase A. Dilute 1.0 mL of
the solution to 10.0 mL with mobile phase A. Di!ute 1.0 mL of Specified impurities: A, B, E.
this solution to 1O.O mL with the test solution. Other detectable impurities (the following substances would,
Column: if present at a sufficient level, be detected by one or other of
- size: l = 0.2S m, 0 = 4.6 mm;
- stationary phase: base-deactivated end-capped octadecylsilyl by the general monograph Substances for pharmaceutical use
silica gel for chromatography R (S µm). (2034). It is therefore not necessary to identify these impurities
Mobile phase: for demonstration of compliance. See also 5.10. Control of
impurities in substances for pharmaceutical use): C, D.
- mobile phase A: mix 3 volumes of methanol R and
97 volumes of a buffer solution prepared as follows:
dissolve 13.6 g of potassium dihydrogen phosphate R in
9SO mL of water for chromatography R, adjust to pH 6.9
with strong sodium hydroxide solution R, add 30 mg of
triethanolamine R and dilute to 1000 mL with water for
chromatography R; A. pyridine-4-carboxylic acid (isonicotinic acid),
- mobile phase B: methanol R;

o - 12 100 o
12 - 20 100 -7 85 o -7 15 B. pyridine-4-carboxamide (isonicotinamide),
20 - 28 85 15
Flow rate: l.S mL/min.

Injection: 10 µL.
C. pyridine-4-carbonitrile (isonicotinonitrile),
with reference solution (b) to identify the peaks due to
impurities A, B and D.
Relative retention with reference to isoniazid (retention
time = about 8.4 min): impurity A = about 0.5;
impurity D = about 1.lS; impurity B = about 1.4.
System suitability: reference solution (b): D. pyridine-3-carbohydrazide (nicotinoyl hydrazide),
- peak-to-valley ratio: minimum 1.8, where HP = height
above the baseline of the peak due to impurity D and
curve separating this peak from the peak due to isoniazid. E. hydrazine.

EUROPEAN PHARMACOPOEIA 9.5 lsotretinoin
07/2018:1019 Límíts:
- ímpurity A: not more than twice the area of the principal
peak in the chromatogram obtained with reference
solution (c) (0.2 per cent);
- unspecifíed ímpurítíes: for each impurity, not more than the
ISOTRETINOIN area of the principal peak in the chromatogram obtained
with reference solution (c) (0.10 per cent);
Isotretinoinum - total: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (c)
(0.5 per cent);
- dísregard limit: 0.5 times the area of the principal peak in
-
the chromatogram obtained with reference solution (c)
(O.OS per cent).
C20H2sÜ2 Mr 300.4
[4759-48-2] Loss on drying (2.2.32): maximum 0.5 per cent, determined
on 1.000 g by drying in vacuo for 16 h.
DEFINITION
(2Z,4E,6E,8E)-3,7-Dimethyl-9-(2,6,6-trimethylcyclohex-1-en- 1.0 g.
1-yl)nona-2,4,6,8-tetraenoic acid.
Content: 98.0 per cent to 102.0 per cent (dried substance). ASSAY
Dissolve 0.200 g in 70 mL of acetone R. Titrate with 0.1 M
CHARACTERS tetrabutylammoníum hydroxide in 2-propanol, determining
Appearance: yellow or light orange, crystalline powder. the end-point potentiometrically (2.2.20).
Solubility: practically insoluble in water, soluble in methylene 1 mL of 0.1 M tetrabutylammonium hydroxide in 2-propanol
--
chloride, slightly soluble in ethanol (96 per cent). is equivalent to 30.04 mg of C20 H 28 0 2•
It is sensitive to air, heat and light, especially in solution.
STORAGE
IDENTIFICATION Under an inert gas, in an airtight container, protected from
light.
-
Infrared absorption spectrophotometry (2.2.24). It is recommended that the contents of an opened container
Comparison: isotretinoin CRS. be used as soon as possible and any unused part be protected
by an atmosphere of inert gas.
TESTS IMPURITIES
Related substances. Liquid chromatography (2.2.29). Carry Specified impurities: A.
out the test protected from light and prepare the solutions
immediately befare use.
Test solution. Dissolve 0.100 g of the substance to be examined th~ tests in the monograph. They are limited by the general
in methanol R and dilute to SO.O mL with the same solvent. acceptance criterion for other/unspecified impurities and/or
Reference solution (a). Dissolve 5.0 mg of tretinoin CRS by the general monograph Substances for pharmaceutical
(impurity A) in methanol R and dilute to 5.0 mL with the use (2034). It is therefore not necessary to identify these
same solvent. impurities for demonstration of compliance. See also 5.1 O.
Reference solution (b). Mix 0.5 mL of the test solution and Control of impurities in substances for pharmaceutical use):
1 mL ofreference solution (a) and dilute to 25 mL with B, C, D, F, G, H, J.
methanol R.
Reference solution (c). Dilute 1.0 mL of the test solution to
-
100.0 mL with methanol R. Dilute 1.0 mL of this solution to
10.0 mL with methanol R.
Column: A. (2E,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-
- size: l = 0.15 m, 0 = 4.6 mm; en-1-yl)nona-2,4,6,8-tetraenoic acid (tretinoin),
- stationary phase: octadecylsilyl silíca gel for
H~co,H
Mobile phase: glacial acetic acid R, water for chromatography R,
methanol R (5:225:770 V/V/V).
Detection: spectrophotometer at 355 nm. H3c
Jnjection: 10 µL of the test solution and reference solutions (b) B. ( 2Z,4E, 6Z,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-
and (c). en-1-yl)nona-2,4,6,8-tetraenoic acid (9,13-dicis-retinoic
Run time: 1.6 times the retention time of isotretinoin. acid),
with reference solution (b) to identify the peak due to
impurity A.
Relative retention with reference to isotretinoin (retention
time= about 26 min): impurity A= about 1.3.
System suitability: reference solution (b): C. (2Z,4Z,6E,8E)-3, 7-dimethyl-9-(2,6,6-trimethylcyclohex-1-
- resolution: minimum 5.0 between the peaks due to en-1-yl)nona-2,4,6,8-tetraenoic acid ( 11,13-dicis-retinoic
isotretinoin and impurity A. acid),
Isotretinoin EUROPEAN PHARMACOPOEIA 9.5
H~ H~O,H
o
C02H
H. (2Z,4E,6E,8E)-3,7-dimethyl-9-(2,6,6-trimethyl-3-
D. (2E,4E,6Z,8E)-3,7-dimethyl-9-(2,6,6-trimethylcyclohex-l- oxocyclohex-l-en-l-yl)nona-2,4,6,8-tetraenoic acid
en-l-yl)nona-2,4,6,8-tetraenoic acid (9-cis-retinoic acid), (13-cis-4-oxoretinoic acid),
H~
C02H
F. (2E,4Z,6E,8E)-3, 7-dimethyl-9-(2,6,6-trimethylcyclohex- l -
H3C
en- l-yl)nona-2,4,6,8-tetraenoic acid (11-cis-retinoic acid),
CH 3 CH3 CH3 H~O,H aodeoaotiome'
(€~~ CH 3
C02H
andenantiomer
H OH
G. (2Z,4E,6E,8E)-3,7-dimethyl-9-[(1RS,6SR)-2,2,6-trimethyl- I. (2Z,4E,6E,8E)-9-[(3RS)-3-hydroxy-2,6,6-
7-oxabicyclo [4.l.O]heptan-l-yl]nona-2,4,6,8-tetraenoic trimethylcyclohex-l-en-l-yl ]-3, 7-dimethylnona-
acid (13-cis-5,6-dihydro-5,6-epoxyretinoic acid), 2,4,6,8-tetraenoic acid (13-cis-4-hydroxyretinoic acid).

L
Lacosamide .............................................................................. 5709 Lactulose, liquid ...................................................................... 5712
Lactulose .................................................................................. 5710

EUROPEAN PHARMACOPOEIA 9.5 Lacosamide
07/2018:2992 Flow rate: l.O mL/min.

Injection: 20 µL of the test solution and reference solutions (b)
and (c).
Run time: l. 7 times the retention time oflacosamide.
LACOSAMIDE Identification of impurities: use the chromatogram obtained
with reference solution (b) to identify the peak due to
Lacosamidum impurity A.
Relative retention with reference to lacosamide (retention
time = about 25 min): impurity A = about 0.8.
impurity A and lacosamide.
C13H1sN203 Mr 250.3 Limit:
[175481-36-4] - impurity A: maximum 0.15 per cent;
- reporting threshold: 0.05 per cent (reference solution (c)).
DEFINITION
(2R)-2-Acetamido-N-benzyl-3-methoxypropanamide.
examined in 1 mL of methanol R and dilute to 10.0 mL with
CHARACTERS water R.
Appearance: white or almost white or light yellow powder. Reference solution (a). Dissolve 50.0 mg of lacosamide CRS in
1 mL of methanol R and dilute to 10.0 mL with water R.
Solubility: sparingly soluble in water, freely soluble in
methanol, practically insoluble in heptane. Reference solution (b ). Dilute 1.0 mL of the test solution to
100.0 mL with a mixture of 10 volumes of methanol R and
lt shows polymorphism (5.9). 90 volumes of water R. Dilute 1.0 mL of this solution to
10.0 mL with the same mixture of solvents.
IDENTIFICATION
Reference solution (e). Disso1ve 5 mg of lacosamide for system
Carry out either tests A, B or tests B, C. suitability CRS (containing impurities B, C, G and I) in 0.1 mL
A. Specific optical rotation (2.2.7): + 14 to + 18 (anhydrous of methanol R and dilute to 1.0 mL with water R.
substance ), measured at 25 ºC. Reference solution (d). Dissolve 1 mg of lacosamide
Dissolve 0.100 gin methanol R and dilute to 10.0 mL with impurity F CRS in 2 mL of methanol R and dilute to 1O.O mL
the same solvent. with water R. Dilute 1.0 mL of the solution to 1O.O mL with
B. Infrared absorption spectrophotometry (2.2.24). water R.
Comparison: lacosamide CRS. Column:
If the spectra obtained in the solid state show differences, - size: i = 0.15 m, 0 = 4.6 mm;
dissolve the substance to be examined and the reference - stationary phase: end-capped extra-dense bonded octylsilyl
substance separately in methanol R, evaporate to dryness silica gel for chromatography R (3.5 µm).
and record new spectra using the residues. Mobile phase:
C. Enantiomeric purity (see Tests). - mobile phase A: 0.1 per cent V/V solution of trifluoroacetic
acid R;
TESTS
- mobile phase B: trifluoroacetic acid R, acetonitrile R,
Appearance of solution. The so1ution is dear (2.2.1) and methanol R (0.3:500:500 V/V/V);
Dissolve 0.500 g in water R and dilute to 50.0 mL with the
same solvent.
Enantiomeric purity. Liquid chromatography (2.2.29): use
o-2 89 11
the normalisation procedure. 2 - 14.2 89 7 69 11 7 31

Test solution. Dissolve 50.0 mg of the substance to be 14.2 - 19.5 69 7 23 31 7 77
examined in the mobile phase and dilute to 50.0 mL with the
mobile phase. 19.5 - 20 23 7 o 77 7 100
Reference solution (a). Dissolve 1 mg of lacosamide 20 - 21 o 100

impurity A CRS in the mobile phase and dilute to 10.0 mL
with the mobile phase. Flow rate: 1.2 mL/mín.
Reference solution (b ). Dissolve 20 mg of the substance to Detection: spectrophotometer at 258 nm.
be examined in the mobile phase, add 1.0 mL ofreference Injection: 20 µL of the test solution and reference solutions (b ),
solution (a) and dilute to 20.0 mL with the mobile phase. (e) and (d).
Reference solution (e). Dilute 1.0 mL of the test solution to Identification of impurities: use the chromatogram
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution supplied with lacosamide for system suitability CRS and
to 20.0 mL with the mobile phase. the chromatogram obtained with reference solutíon (e) to
identify the peaks due to impuríties B, C, G and I; use the
Column:
chromatogram obtained with reference solution (d) to identify
- size: l = 0.25 m, 0 = 4.6 mm; the peak due to impurity F.
- stationary phase: amylose derivative of silica gel for chiral Relative retention with reference to lacosamide
separation R (10 µm). (retention time = about 12 min): impuríty F = about
Mobile phase: water for chromatography R, 2-propanol R, O. 7; impurity G = about 0.9; impurity B = about 1.2;
heptane R (3:100:900 V/V/V). impurity C = about 1.3; ímpurity I = about 1.6.
Lactulose EUROPEAN PHARMACOPOEIA 9.5
System suitability: reference solution (c):

impurity G and lacosamide; minimum 4.5 between the
peaks due to lacosamide and impurity B.
Calculation of percentage contents: D. (22)-2-amino-N-benzyl-3-methoxypropanamide,
- correction factors: multiply the peak areas of the following
impurity G = 0.7; impurity I = 0.7;
- for each impurity, use the concentration of lacosamide in
reference solution (b ).
Limits:
- impurities B, C, F, G, I: for each impurity, maximum
0.15 per cent;
0.10 per cent;
- total: maximum 0.5 per cent; F. (22)-2-acetamido-N-benzyl-3-hydroxypropanamide,
Water (2.5.32): maximum 0.2 per cent, determined on 0.150 g
by direct sample introduction.
H,Cy~»
o
1.0 g.
ASSAY
related substances with the following modifications.
System suitability: reference solution (a): H. (22)-2-acetamido-N-[(22)-l-(benzylamino)-3-methoxy-l-
- symmetry factor: maximum 2.4 for the peak due to oxopropan-2-yl ]-3-methoxypropanamide,
lacosamide.
Calculate the percentage content of C13 H 18N 20 3 taking into
account the assigned content of lacosamide CRS.
O
N)l_N
H
r--
H~
OCH3»
H
N
1
#
() o
IMPURITIES
Specified impurities: A, B, C, F, G, I. I. (22)-N-benzyl-2-[ (benzylcarbamoyl)amino ]-3-
Other detectable impurities (the following substances would, methoxypropanamide,
acceptance criterion for other/unspecified impurities and/or H2N»
(2034). It is therefore not necessary to identify these impurities J. phenylmethanamine,
impurities in substances for pharmaceutical use): D, E, H, J, K. O CH2H 0
HCANJl ,NJV
3 H- "
o
K. 2-acetamido-N-benzylprop-2-enamide.
A. (2S)-2-acetamido-N-benzyl-3-methoxypropanamide, m
6
07/2018:1230
LACTULOSE
Lactulosum
B. (22)-2-acetamido-3-(benzylamino )-3-oxopropyl aceta te,

HO~~JOH
H0~')-3''zoH
HO O O
OH
OH
C. (22)-N-benzyl-3-methoxy-2-(N-methylacetamido)- Mr 342.3
propanamide,

EUROPEAN PHARMACOPOEIA 9.S Lactulose
DEFINITION Reference solution (b ). Dissolve 1.00 g of lactulose CRS in

4-0-~- D-Galactopyranosyl-D-arabino-hex-2-ulofuranose.
10 mL of water R. Add 12.5 mL of acetonitrile R with gentle
heating and dilute to 2S.0 mL with water R.
Content: 9S.O per cent to 102.0 per cent (anhydrous substance).
Reference solution (e). Dissolve 10 mg of lactulose R, 10 mg of
CHARACTERS epilactose R (impurity A) and 10 mg of lactase monohydrate R
Appearance: white or almost white, crystalline powder. (impurity C) in 2.0 mL of water R. Add 2.S mL of acetonitrile R
with gentle heating and dilute to S.O mL with water R.
Solubility: freely soluble in water, sparingly soluble in
methanol, practically insoluble in toluene. Reference solution (d). To S.O mL of the test solution add
47.S mL of acetonitrile R with gentle heating and dilute to
mp: about 168 ºC. 100.0 mL with water R. Dilute S.0 mL of this solution to
IDENTIFICATION 100.0 mL with a mixture of equal volumes of acetonitrile R
and water R.
First identification: B, C, D, E.
Column 1:
Second identification: A, C, D, E.
- size: l =O.OS m, 0 = 4.6 mm;
A. Thin-layer chromatography (2.2.27).
- stationary phase: aminopropylsilyl silíca gel for
Test solution. Dissolve SO.O mg of the substance to be chromatography R (3 µm);
examined in water R and dilute to 1O.O mL with the same
solven t. - temperature: 38 ± 1 ºC.
Reference solution. Dissolve SO.O mg of lactulose CRS in Column 2:
water R and dilute to 10.0 mL with the same solvent. - size: l = O.lS m, 0 = 4.6 mm;
Plate: TLC silica gel plate R. - stationary phase: aminopropylsilyl silica gel for
Mobile phase: glacial acetic acid R, SO g/L solution of boric
acid R, methanol R, ethyl acetate R {10:1S:20:SS V/V/V/V). - temperature: 38 ± 1 ºC.
Application: 2 µL. Columns 1 and 2 are coupled in series.
Development: over 3/4 of the plate. Mobile phase: dissolve 0.2S3 g of sodium dihydrogen
phosphate R in 200 mL of water for chromatography R and
Drying: at 100-lOS ºC for S min; allow to cool. dilute to 1000 mL with acetonitrile R.
Detection: spray with a 1.0 g/L solution of Flow rate: 1.0 mL/min.
1,3-dihydroxynaphthalene R in a mixture of 10 volumes
of sulfuric acid R and 90 volumes of methanol R; heat at Detection: refractometer maintained at a constant temperature.
110 ºC for S min. Injection: 20 µL of the test solution and reference solutions (a),
(c) and (d).
Results: the principal spot in the chromatogram obtained
with the test solution is similar in position, colour and size Run time: twice the retention time of lactulose.
to the principal spot in the chromatogram obtained with Identification of impurities: use the chromatogram obtained
the reference solution. with reference solution (c) to identify the peaks dueto
B. Examine the chromatograms obtained in the assay. impurities A and C.
Results: the principal peak in the chromatogram obtained Relative retention with reference to lactulose (retention
with the test solution is similar in retention time and size time== about 18 min): impurity A= about 0.9;
to the principal peak in the chromatogram obtained with impurity e = about 1.2.
reference solution (b ). System suitability: reference solution (c):
C. Dissolve SO mg in 1O mL of water R. Add 3 mL of - peak-to-valley ratio: minimum S.O, where HP = height
cupri-tartaric solution R and heat. A red precipitate is above the baseline of the peak due to impurity A and
formed. Hv = height above the baseline of the lowest point of the
D. Dissolve 0.12S gin S mL of water R. Add S mL of curve separating this peak from the peak due to lactulose.
ammonia R. Heat in a water-bath at 80 ºC for 10 min. A Limits:
red colour develops. - impurity C: not more than the area of the peak due to
E. Specific optical rotation (see Tests). lactulose in the chromatogram obtained with reference
solution (a) (3.0 per cent);
TESTS - unspecified impurities: for each impurity, not more
Solution S. Dissolve 3.0 gin carbon dioxide-free water R and than twice the area of the peak due to lactulose in the
dilute to SO mL with the same solvent. chromatogram obtained with reference solution (d) (0.5 per
Appearance of solution. Solution S is clear (2.2.1) and not cent);
more intensely coloured than reference solution BY 5 (2.2.2, - total: not more than the area of the peak due to lactulose
Method JI). in the chromatogram obtained with reference solution (a)
(3.0 per cent);
pH (2.2.3): 3.0 to 7.0.
- disregard limit: the area of the peak dueto lactulose in
To 10 mL of solution S add 0.1 mL of a saturated solution of the chromatogram obtained with reference solution (d)
potassium chloride R. (0.2S per cent).
Specific optical rotation (2.2.7): - SO.O to - 46.0 (anhydrous The thresholds indicated under Related substances
substance). (Table 2034.-1) in the general monograph Substances for
Dissolve l.2S g in water R, add 0.2 mL of concentrated pharmaceutical use (2034) do not apply.
ammonia R and dilute to 2S.O mL with water R. Methanol. Head-space gas chromatography (2.2.28).
Related substances. Liquid chromatography (2.2.29). Interna[ standard solution. Mix O.S mL of propano[ R and
Test solution. Dissolve 1.00 g of the substance to be examined 100.0 mL of water R. Dilute 1.0 mL of the solution to 100.0 mL
in 10 mL of water R. Add 12.S mL of acetonitrile R with gentle with water R. Dilute S.0 mL of this solution to SO.O mL with
heating and dilute to 2S.O mL with water R. water R.
Reference solution (a). To 3.0 mL of the test solution add Test solution. To 79 mg of the substance to be examined in a
48.S mL of acetonitrile R with gentle heating and dilute to 20 mL vial add 1.0 mL of the intemal standard solution and
100.0 mL with water R. S µL of a 0.1 per cent V/V solution of methanol R.
Lactulose, liquid EUROPEAN PHARMACOPOEIA 9.S
Reference solution. To 1.0 mL of the interna! standard solution Calculate the percentage content of C 12 H 22 0 11 taking into
in a 20 mL vial add S µL of a 0.1 per cent V/V solution of account the assigned content of lactulose CRS.
methanol R.
IMPURITIES
Column:
Specified impurities: C.
- size: l = 2 m, 0 = 2 mm;
- stationary phase: ethylvinylbenzene-divinylbenzene if present ata sufficient leve!, be detected by one or other of
copolymer R (180 µm). the tests in the monograph. They are limited by the general
Carrier gas: helium for chromatography R. acceptance criterion for other/unspecified impurities. It
Flow rate: 30 mL/min. is therefore not necessary to identify these impurities for
Static head-space conditions that may be used: demonstration of compliance. See also 5.1 O. Control of
- equilibration temperature: 60 ºC; impurities in substances for pharmaceutical use): A, B, D, E.
HO~HO~~
- equilibration time: 1 h;
- pressurisation time: 1 min.
O~
Temperature: OH
- column: 140 ºC; HO

- injection port: 200 ºC; OH
- detector: 220 ºC.
Detection: flame ionisation. OH
Injection: 1 mL of the gaseous phase. A. 4-0- ~- D-galactopyranosyl-D-mannopyranose (epilactose ),

Calculate the content of methanol, taking its density (2.2.5) at
20 ºC to be 0.79 g/mL.
Limit:
- methanol: calculate the ratio (R) of the area of the peak
:;;_o,
\L(OH
due to methanol to the area of the peak due to the interna!
standard in the chromatogram obtained with the reference OH
solution; calculate the ratio of the area of the peak due
to methanol to the area of the peak due to the interna! B. D-galactopyranose (galactose),
standard in the chromatogram obtained with the test
solution: this ratio is not greater than 2R (SO ppm). HOI -
HO~~u~OH
Boron: maximum 9 ppm.
A void where possible the use of glassware.
Reference solution. Dissolve SO.O mg of boric acid R in water R HO
OH
O ?X-f OH
and dilute to 100.0 mL with the same solvent. Dilute S.O mL
of the solution to 100.0 mL with water R. Keep in a well-closed
polyethylene container. OH
In 4 polyethylene 2S mL flasks, place separately: C. 4-0-~-D-galactopyranosyl-D-glucopyranose (lactose ),

- O.SO g of the substance to be examined dissolved in 2.0 mL
of water R (solution A); ~~y-OH
- O.SO g of the substance to be examined dissolved in 1.0 mL H¿)-Y' 'oH
of the reference solution and 1.0 mL of water R (solution B);
OH
- 1.0 mL of the reference solution and 1.0 mL of water R
(solution C); D. D-arabino-hex-2-ulopyranose (fructose),
- 2.0 mL of water R (solution D).
To each flask add 4.0 mL of acetate-edetate buffer solution
pH 5.5 R. Mix and add 4.0 mL of freshly prepared
azomethine H solution R. Mix and allow to stand for 1 h.
Measure the absorbance (2.2.25) of solutions A, B and C at E. D-lyxo-hex-2-ulopyranose (tagatose).
420 nm, using solution D as the compensation liquid. The
test is not valid unless the absorbance of solution C is at least 07/2018:0924
0.2S. The absorbance of solution B is not less than twice that
of solution A.
Lead (2.4.1 O): maximum 0.5 ppm.
Water (2.5.12): maximum 2.5 per cent, determined on 0.500 g.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on LACTULOSE, LIQUID
1.0 g.
Microbial contamination Lactulosum liquidum
TAMC: acceptance criterion 102 CFU/g (2.6.12). DEFINITION
Absence of Escherichia coli (2.6.13). Aqueous solution of 4-0-~-D-galactopyranosyl-D
arabino-hex-2-ulofuranose normally prepared by alkaline
ASSAY isomerisation of lactose. It may contain other sugars including
Liquid chromatography (2.2.29) as described in the test for lactose, epilactose, galactose, tagatose and fructose.
related substances with the following modifications. Content: minimum 620 g/L oflactulose (C 12 H 22 Ü 11 ; Mr 342.3)
Injection: test solution and reference solution (b). and 95.0 per cent to 105.0 per cent of the content oflactulose
System suitability: reference solution (b): stated on the label.
- symmetry factor: 0.6 to 2.0 for the principal peak. It may contain a suitable antimicrobial preservative.

EUROPEAN PHARMACOPOEIA 9.S Lactulose, liquid
CHARACTERS Reference solution (e). Dilute S.O mL of reference solution (a)

Appearance: clear, viscous liquid, colourless or pale to 100.0 mL with a mixture of equal volumes of acetonitrile R
brownish-yellow. and water R.
Solubility: miscible with water. It may be a supersaturated Column 1:
solution or may contain crystals that disappear on heating. - size: l =O.OS m, 0 = 4.6 mm;
A 1O per cent V/V solution is laevorotatory. - stationary phase: aminopropylsilyl silica gel for
IDENTIFICATION chromatography R (3 µm);
First identification: B, C, D. - temperature: 38 ± 1 ºC.
Second identification: A, C, D. Column 2:
A. Thin-layer chromatography (2.2.27). - size: l = O.lS m, 0 = 4.6 mm;
Test solution. Dilute O.SO g of the substance to be examined - stationary phase: aminopropylsilyl silica gel for
in water R and dilute to SO.O mL with the same solvent. chromatography R (3 µm);
Reference solution. Dissolve 60.0 mg of lactulose CRS in - temperature: 38 ± 1 ºC.
water R and dilute to 10.0 mL with the same solvent. Columns 1 and 2 are coupled in series.
Plate: TLC silica gel plate R. Mobile phase: dissolve 0.2S3 g of sodium dihydrogen
Mobile phase: glacial acetic acid R, SO g/L solution of boric phosphate R in 200 mL of water for chromatography R and
acid R, methanol R, ethyl acetate R (10:1S:20:SS V/VIV!V). dilute to 1000 mL with acetonitrile R.
Application: 2 µL. Flow rate: 1.0 mL/min.
Development: over 3/4 of the plate. Detection: refractometer maintained at a constant temperature.
Drying: at 100-lOS ºC for S min and allow to cool. Injection: 20 µL of the test solution and of reference
Detection: spray with a 1.0 g/L solution of solutions (a), (d) and (e).
1,3-dihydroxynaphthalene R in a mixture of 10 volumes
Run time: twice the retention time of lactulose.
of sulfuric acid R and 90 volumes of methanol R; heat at
110 ºC for S min. Identification of impurities: use the chromatogram
Results: the principal spot in the chromatogram obtained supplied with lactulose for peak identification CRS and the
with the test solution is similar in position, colour and size chromatogram obtained with reference solution (d) to identify
to the principal spot in the chromatogram obtained with the peaks due to impurities A, B, C, D, E, F, G and H.
the reference solution. Relative retention with reference to lactulose (retention
B. Examine the chromatograms obtained in the assay. time = about 18 min): impurity F = about 0.2;
impurity E = about 0.38; impurity D = about 0.42;
Results: the principal peak in the chromatogram obtained
impurity B = about 0.6; impurity G = about 0.8;
with the test solution is similar in retention time to
impurity A = about 0.9; impurity C = about 1.2;
the principal peak in the chromatogram obtained with
impurity H = about 1.5.
reference solution (b ).
System suitability: reference solution (d):
C. To 0.1 g add 10 mL of water R and 3 mL of cupri-tartaric
4
solution R and heat. ,.. red precipitate is formed.
.1.
- peak-to-valley ratio: minimum S.O, where HP = height
D. To 0.2S g add S mL of water R and S mL of ammonia R. above the baseline of the peak due to impurity A and
Heat in a water-bath at 80 ºC for 10 min. A red colour Hv = height above the baseline of the lowest point of the
develops. curve separating this peak from the peak due to lactulose.
Limits:
TESTS
- impurity B: not more than 3 times the area of the peak due
Solution S. Mix 1O g with carbon dioxide-free water R and to lactulose in the chromatogram obtained with reference
dilute to 100 mL with the same solvent. solution (a) (IS.O per cent);
Appearance of solution. Solution S is clear (2.2.1) and not - impurities A, C: for each impurity, not more than twice
more intensely coloured than reference solution BY 5 (2.2.2, the area of the peak due to lactulose in the chromatogram
Method JI). obtained with reference solution (a) (10.0 per cent);
pH (2.2.3): 3.0 to 7.O. - impurities E, F: for each impurity, not more than 0.8 times
To 10 mL of solution S add 0.1 mL of a saturated solution of the area of the peak due to lactulose in the chromatogram
potassium chloride R. obtained with reference solution (a) (4.0 per cent);
Related substances. Liquid chromatography (2.2.29). - impurities G, H: for each impurity, not more than 0.3 times
Test solution. Mix 4.00 g of the substance to be examined and the area of the peak due to lactulose in the chromatogram
20 mL of water R. Add 2S.O mL of acetonitrile R with gentle obtained with reference solution (a) (l.S per cent);
heating and dilute to SO.O mL with water R. - impurity D: not more than 0.2 times the area of the peak
Reference solution (a). To S.O mL of the test solution add due to lactulose in the chromatogram obtained with
47.5 mL of acetonitrile R with gentle heating and dilute to reference solution (a) (LO per cent);
100.0 mL with water R. - unspecified impurities: for each impurity, not more than
Reference solution (b ). Dissolve 2.00 g of lactulose CRS in 0.1 times the area of the peak dueto lactulose in the
20 mL of water R. Add 2S.O mL of acetonitrile R with gentle chromatogram obtained with reference solution (a) (0.5 per
heating and dilute to SO.O mL with water R. cent);
Reference solution (e). Dissolve 6S mg of fructose CRS - sum of impurities eluting after impurity H: not more than
(impurity D) in a mixture of equal volumes of acetonitrile R 0.26 times the area of the peak due to lactulose in the
and water R and dilute to 100.0 mL with the same mixture chromatogram obtained with reference solution (a) (1.3 per
of solvents. cent);
Reference solution (d). Dissolve 1 g of lactulose for peak - total (excluding impurities B and C): not more than
identification CRS (containing impurities A, B, C, E, F, G 2.4 times the area of the peak due to lactulose in the
and H) in reference solution (c) and dilute to 2S.O mL with chromatogram obtained with reference solution (a)
reference solution (c). (12.0 per cent);
Lactulose, liquid EUROPEAN PHARMACOPOEIA 9.5
- disregard limit: not more than the area of the peak due to In 4 polyethylene 25 mL flasks, place separately:
lactulose in the chromatogram obtained with reference - 1.00 g of the substance to be examined and 1 mL of water R
solution (e) (0.25 per cent). (solution A);
The thresholds indicated under Related substances - 1.00 g of the substance to be examined and 1 mL of the
(Table 2034.-1) in the general monograph Substances for reference solution (solution B);
pharmaceutical use (2034) do not apply. - 1 mL of the reference solution and 1 mL of water R
Methanol. Head-space gas chromatography (2.2.28). (solution C);
Interna[ standard solution. Mix 0.5 mL of propano[ R and - 2 mL of water R (solution D).
100.0 mL of water R. Dilute 1.0 mL of the solution to 100.0 mL To each flask, add 4.0 mL of acetate-edetate buffer solution
with water R. Dilute 5.0 mL of this solution to SO.O mL with pH 5.5 R. Mix and add 4.0 mL of freshly prepared
water R. azomethine H solution R. Mix and allow to stand for 1 h.
Test solution. To 0.13 g of the substance to be examined in a Measure the absorbance (2.2.25) of solutions A, B and C at
20 mL vial add 1.0 mL of the interna! standard solution and 420 nm, using solution D as the compensation liquid. The
5 µL of a 0.1 per cent V/V solution of methanol R. test is not valid unless the absorbance of solution C is at least
0.25. The absorbance of solution B is not less than twice that
Reference solution. To 1.0 mL of the interna! standard solution of solution A.
in a 20 mL vial add 5 µL of a 0.1 per cent V/V solution of
methanol R. Lead (2.4.1 O): maximum 0.5 ppm, calculated with reference
to the declared content of lactulose.
Column:
- size: l = 2 m, 0 = 2 mm; 1.5 g and calculated with reference to the declared content of
- stationary phase: ethylvinylbenzene-divinylbenzene lactulose.
copolymer R (180 µm). Microbial contamination
Carrier gas: helium for chromatography R. TAMC: acceptance criterion 102 CFU/g (2.6.12).
Flow rate: 30 mL/min. TYMC: acceptance criterion 10 1 CFU/g (2.6.12).
Static head-space conditions which may be used: Absence of Escherichia coli (2.6.13).
- equilibration temperature: 60 ºC; ASSAY
- equilibration time: 1 h; Liquid chromatography (2.2.29) as described in the test for
- pressurisation time: 1 min. related substances with the following modifications.
Temperature: Injection: test solution and reference solution (b).
- column: 140 ºC; System suitability: reference solution (b):
- symmetry factor: 0.6 to 2.0 for the principal peak.
- injection port: 200 ºC;
Calculate the percentage content of C12 H 22 0 11 taking into
- detector: 220 ºC. account the assigned content of lactulose CRS.
LABELLING
Injection: 1 mL of the gaseous phase.
The label states the declared content of lactulose.
Calculate the content of methanol, taking its density (2.2.5) at
20 ºC to be 0.79 g/ml. IMPURITIES
Limit: Specified impurities: A, B, C, D, E, F, G, H.
- methanol: calculate the ratio (R) of the area of the peak
due to methanol to the area of the peak due to the interna!
standard in the chromatogram obtained with the reference
solution; calculate the ratio of the area of the peak due
HOt?HO~~
O~
OH
to methanol to the area of the peak due to the interna! HO

standard in the chromatogram obtained with the test OH
solution: this ratio is not greater than 2R (30 ppm).
Sulfites: maximum 30 ppm. OH
Mix 5.0 g with 40 mL of water R, add 2.0 mL of 0.1 M sodium A. 4-0-~- D-galactopyranosyl-D-mannopyranose (epilactose ),
hydroxide and dilute to 100 mL with water R. To 10.0 mL
of this solution, add 1.0 mL of hydrochloric acid Rl, 2.0 mL
of decolorised fuchsin solution Rl and 2.0 mL of a 0.5 per
cent V/V solution offormaldehyde R. Allow to stand for 30 min
and measure the absorbance (2.2.25) at 583 nm using as the
:;;_o,
\L(OH
compensation liquid a solution prepared at the same time and
in the same manner with 10.0 mL of water R instead of the OH
solution of the substance to be examined. The absorbance is
B. D-galactopyranose (galactose),
not greater than that of a reference solution prepared at the
same time and in the same manner using 10.0 mL of sulfite
standard solution (1.5 ppm SOJ R instead of the solution of
the substance to be examined. HOt?HO~O, OH
Boron: maximum 5 ppm.
A void where possible the use of glassware.
HO O~
OH OH
Reference solution. Dissolve 56.0 mg of boric acid R in water R
and dilute to 100.0 mL with the same solvent. Dilute 5.0 mL of OH
this solution to 100.0 mL with water R. Keep in a well-closed
polyethylene container. C. 4-0-~-D-galactopyranosyl-n-glucopyranose (lactose),

EUROPEAN PHARMACOPOEIA 9.5 Lactulose, liquid
O rOH
HO yº"
D. D-arabino-hex-2-ulopyranose (fructose), F. (4~)-3-deoxypent-2-ulofuranose,
G. unknown structure,
E. D-lyxo-hex-2-ulopyranose (tagatose), H. unknown structure.

M
Molgramostim concentrated solution .................................. 5719 Mometasone furoate monohydrate ...................................... 5724
Mometasone furoate ............................................................... 5721

EUROPEAN PHARMACOPOEIA 9.5 Molgramostim concentrated solution
01/2008:1641 and 268 volumes of water R (mixture A) and rinse for

corrected 9.5 5 min. Immerse the gel for 1O min in a staining solution
prewarmed to 60 ºC and prepared by adding acid blue
83 R at a concentration of 1.2 g/L to mixture A. Wash the
gel in severa! containers with mixture A and keep the gel
in this mixture until the background is clear (12-24 h).
MOLGRAMOSTIM CONCENTRATED After adequate destaining, soak the gel for 1 h in a 10 per
cent V/V solution of glycerol R in mixture A.
SOLUTION System suitability:
Molgramostimi solutio concentrata - in the electropherogram obtained with reference
solution (b), the relevant isoelectric point markers are
APARSPSPST QPWEHVNAIQ EARRLLNLSR
distributed along the entire length of the gel;
- in the electropherogram obtained with reference
DTAAEMNETV EVISEMFDLQ EPTCLQTRLE solution (a), the pl of the principal band is 4.9 to 5.4.
LYKQGLRGSL TKLKGPLTMM ASHYKQHCPP Results: the principal band in the electropherogram
obtained with the test solution corresponds in position
TPETSCATQI ITFESFKENL KDFLLVIPFD to the principal band in the electropherogram obtained
CWEPVQE with reference solution (a). Plot the migration distances of
the relevant pl markers versus their pl and determine the
c639H1007N1710196ss Mr 14 477 isoelectric points of the principal component of each of the
test solution and reference solution (a). They do not differ
DEFINITION
by more than 0.2 pI units.
Solution of a protein having the structure of the granulocyte C. Examine the electropherograms obtained under reducing
macrophage colony stimulating factor which is produced conditions in the test for impurities with molecular masses
and secreted by various human blood cell types. The protein differing from that of molgramostim. The principal band
stimulates the differentiation and proliferation of leucocyte in the electropherogram obtained with test solution (a)
stem cells into mature granulocytes and macrophages. is similar in position to the principal band in the
Content: minimum 2.0 mg of protein per millilitre. electropherogram obtained with reference solution (a).
Potency: minimum 0.7 x 107 IU per milligram of protein. D. Peptide mapping (2.2.55).
PRODUCTION Test solution. Introduce 50 µL of tris-hydrochloride
Molgramostim concentrated solution is produced by a buffer solution pH 8.0 R and 50 µL of the preparation
method based on recombinant DNA (rDNA) technology, to be examined at a concentration of 2 mg/mL into a
using bacteria as host cells. It is produced under conditions polypropylene tube of 0.5 mL capacity. Add 4 µL of a
1 mg/mL solution of trypsin for peptide mapping R in a
designed to minimise microbial contamination of the product.
0.01 per cent V/V solution of trifluoroacetic acid R, cap
Prior to release, the following tests are carried out on each tightly and mix well. Incubate at about 37 ºC for 18 h.
batch of the final bulk product, unless exemption has been Add 125 µL of a 764 g/L (8 M) solution of guanidine
granted by the competent authority. hydrochloride R and mix well. Add 10 µL of a 154.2 g/L
Host-cell derived proteins: the limit is approved by the (1 M) solution of dithiothreitol R and mix well. Place the
competent authority. capped tube in boiling water for 1 min. Cool to room
Host-cell or vector derived DNA: the limit is approved by temperature.
the competent authority. Reference solution. Prepare at the same time and in the same
manner as for the test solution but use molgramostim CRS
CHARACTERS instead of the preparation to be examined.
Appearance: clear, colourless liquid. Examine the 2 tryptic digests by liquid chromatography
(2.2.29).
IDENTIFICATION
Column:
A. It shows the expected biological activity (see Assay).
- size: l = 0.10 m, 0 = 4.6 mm;
B. Isoelectric focusing (2.2.54).
- stationary phase: octadecylsilyl silica gel for
Test solution. Dilute the preparation to be examined with chromatography R (5 µm) with a pore size of 30 nm.
water R to obtain a concentration of 0.25 mg/mL.
Mobile phase:
Reference solution (a). Dilute molgramostim CRS with
water R to obtain a concentration of 0.25 mg/mL. - mobile phase A: dilute 1 mL of trifluoroacetic acid R in
1000 mL of water f or chromatography R;
Reference solution (b). Use an isoelectric point (pI)
calibration solution, in the pl range of 2.5-6.5, prepared - mobile phase B: dilute 1 mL of trifluoroacetic acid R in
according to the manufacturer's instructions. 100 mL of water for chromatography R; add 900 mL of
acetonitrile Rl and mix;
Focusing:
- pH gradient: 4.0-6.5; Time Mobile phase A Mobile phase B
catholyte: 8.91 g/L (O.l M) solution of 3-aminopropionic
acid R; o - 35.0 100 -7 65 o -7 35
- anolyte: 14.7 g/L (0.1 M) solution of glutamic acid R 35.0 - 105.0 65 -7 35 35 -7 65
in a 50 per cent V/V solution of dilute phosphoric 105.0 - 107.5 35 -7 100 65 -7 o
acid R (0.5 M);
- application: 20 µL. 107.5 - 120.0 100 o
Detection : immerse the gel in a suitable volume of a solution Flow rate: 1.0 mL/min.
containing 115 g/L of trichloroacetic acid R and 34.5 g/L
of suljosalicylic acid R and shake the container gently for Detection: spectrophotometer at 214 nm.
30 min. Transfer the gel to a mixture of 32 volumes of Equilibration: at initial conditions for at least 12 min.
glacial acetic acid R, 100 volumes of anhydrous ethanol R Injection: 200 µL.
Molgramostim concentrated solution EUROPEAN PHARMACOPOEIA 9.5
System suitability: the chromatograms obtained with the Reference solution (a). Dilute molgramostim CRS in water R
reference solution and the test solution are qualitatively to obtain a concentration of 0.02 mg/mL. Mix 1 volume of
similar to the Ph. Eur. reference chromatogram of this solution with 1 volume of concentrated SDS-PAGE sample
molgramostim digest. buffer R.
Results: the profile of the chromatogram obtained with Reference solution (b). Prepare as for reference solution (a),
the test solution corresponds to that of the chromatogram but using concentrated SDS-PAGE sample buffer for reducing
obtained with the reference solution. conditions R.
E. N- Terminal sequence analysis. Reference solution (c). Use a solution of molecular mass
Perform the Edman degradation using an automated markers suitable for calibrating SDS-PAGE gels in the range
of 14 400-94 000. Dissolve in sample buffer or sample buffer
solid-phase sequencer, operated in accordance with the
(reducing conditions), as appropriate.
manufacturer's instructions.
Sample treatment: boíl for 3 min.
Load about 1 nmol of the test preparation to a sequencing
cartridge using the protocol provided by the manufacturer. Application: SO µL; apply reduced and non-reduced solutions
Run 16 sequencing cycles, noting, if appropriate, the to separate gels.
presence of proline at positions 2, 6, 8 and 12. Detection: silver staining as described below.
Identify the phenylthiohydantoin (PTH)-amino acids Immerse the gel overnight in a mixture of 1Ovolumes of acetic
released at each sequencing cycle by reverse-phase acid R, 40 volumes of water R and SO volumes of methanol R.
liquid chromatography. The procedure may be carried Transfer the gel to a 100 g/L solution of glutaraldehyde R
out using the column and reagents recommended by and shake for about 30 min. Replace the glutaraldehyde
the manufacturer of the sequencing equipment for the solution with water R, and keep the gel in water R for
separation of PTH-amino acids. 20 min. Repeat this washing-step twice. Transfer the gel to a
The separation procedure is calibrated using: mixture containing 0.7S g/L of sodium hydroxide R, 14 g/L
of concentrated ammonia R and 8 g/L of silver nitrate R.
- the mixture of PTH-amino acids provided by the This solution is prepared immediately before use. Place the
manufacturer of the sequencer, with the gradient gel on a shaker in the dark for S min. Wash the gel for 30 s
conditions adjusted as indicated to achieve optimum in each of 3 containers with water R and shake the gel in a
resolution of all amino acids; mixture consisting of O.OS g/L of citric acid monohydrate R,
- a sample obtained from a blank sequencing cycle O.OS per cent V/V of formaldehyde R and O.OOS per cent V/V
obtained as recommended by the equipment of methanol R in water R. Protein bands become visible
manufacturer. during this step. Keep the gel in the solution until sufficiently
Results: the first 16 amino acids are: Ala-Pro-Ala-Arg-Ser- stained and then rinse the gel repeatedly with water R in a
Pro-Ser-Pro-Ser-Thr-Gln-Pro-Trp-Glu-His-Val. shaking water bath. Soak gels in a solution consisting of 10 per
cent V/V of acetic acid R and 1 per cent V/V of glycerol R.
TESTS System suitability:
Impurities with molecular masses differing from that of - the validation criteria are met (2.2.31);
molgramostim. Polyacrylamide gel electrophoresis (2.2.31) - a band is seen in the electropherogram obtained with test
under both reducing and non-reducing conditions. solution (h);
Gel dimensions: 0.7S mm thick. - a gradation of intensity of staining is seen in the
Resolving gel: 14 per cent acrylamide. electropherograms obtained with test solutions (a)-(h)
Sample buffer A. Mix equal volumes of water R and and (i)-(p);
concentrated SDS-PAGE sample buffer R. - the molecular mass of the principal band in the
electrophoregram obtained with reference solution (a)
Sample buffer B (reducing conditions). Mix equal volumes
or (b) is within the range of l S 100 to 17 100.
of water R and concentrated SDS-PAGE sample buffer for
reducing conditions R. Limits: compare the staining intensity of each
non-molgramostim band observed in the electropherogram
Test solution (a). Dilute the preparation to be examined in obtained with test solution (a) to the staining intensity
water R to obtain a concentration of 1.0 mg/mL. To 1 volume of the principal band in the electropherograms obtained
of this solution add 1 volume of concentrated SDS-PAGE with test solutions (b)-(h). Proceed similarly with the
sample buffer R. electropherograms obtained with test solutions (i)-(p ). The
Test solution (b) (2 per cent control). Dilute 0.020 mL of test impurity level is estimated as the dilution, in percentage, of
solution (a) to 1.0 mL with sample buffer A. the solution giving the electropherogram with the closest
Test solution (c) (1 per cent control). To 0.20 mL of test intensity of staining.
solution (b) add 0.20 mL of sample buffer A. Reducing conditions:
Test solution (d) (O.S per cent control). To 0.20 mL of test - impurity with an apparent molecular mass of 20 000:
solution (c) add 0.20 mL of sample buffer A. maximum 1 per cent;
Test solution (e) (0.2S per cent control). To 0.20 mL of test - impurity with an apparent molecular mass of 25 000:
solution (d) add 0.20 mL of sample buffer A. maximum 0.1 per cent;
Test solution (f) (0.1 per cent control). To 0.20 mL of test - impurity with an apparent molecular mass of 30 000:
solution (e) add 0.30 mL of sample buffer A. maximum 0.3 per cent;
Test solution (g) (O.OS per cent control). To 0.20 mL of test - total: maximum 2 per cent.
solution (f) add 0.20 mL of sample buffer A. Non-reducing conditions:
Test solution (h) (0.02S per cent control). To 0.20 mL of test - total of all impurities of molecular masses higher than
solution (g) add 0.20 mL of sample buffer A. 30 000: maximum 1 per cent.
Test solution (i). Prepare as for test solution (a), but Related proteins. Liquid chromatography (2.2.29): use the
using concentrated SDS-PAGE sample buffer for reducing normalisation procedure.
conditions R. Test solution (a). Dilute the preparation to be examined
Test solutions (j)-(p). Prepare as for test solutions (b)-(h), but with 0.05 M phosphate buffer solution pH 7.0 R to obtain a
using sample buffer B. concentration of O.S mg/mL.

EUROPEAN PHARMACOPOEIA 9.5 Mometasone furoate
Test solution (b). Mix 1 volume of test solution (a) with International Standard of molgramostim or with a reference
4 volumes of a 0.125 mg/mL solution of human albumin R or preparation calibrated in International Units, which yield the
bovine albumin R in 0.05 M phosphate buffer solution pH 7.0 R. same response (SO per cent maximal stimulation).
Reference solution (a). Dilute molgramostim CRS with 0.05 M The International Unit is the activity contained in a stated
phosphate buffer solution pH 7.0 R to obtain a concentration amount of the appropriate International Standard. The
of 0.5 mg/mL. equivalence in International Units of the International
Reference solution (b). Mix 1 volume of reference solution (a) Standard is stated by the World Health Organization.
with 4 volumes of a 0.125 mg/mL solution of human Add 50 µL of dilution medium to all wells of a 96-well
albumin R or bovine albumin R in 0.05 M phosphate buffer microtitre plate. Add an additional 50 µL of this solution to
solution pH 7.0 R. the wells designed for the blanks. Add 50 µL of each solution
Column: to be tested in triplica te (test preparation and reference
- size: l = 0.15 m, 0 = 4.6 mm; preparation ata concentration of about 65 IU/mL, plus a series
of 10 twofold dilutions to obtain a standard curve). Then
add to each well 50 µL of a TF-1 cell suspension containing
chromatography R (5 µm) with a pore size of 30 nm.
3 x 10 5 cells per millilitre, maintaining the cells in a uniform
Mobile phase: suspension during addition.
- mobile phase A: to about 800 mL of water for Incubate the plate at 36.0-38.0 ºC for a minimum of 24 h in a
chromatography R add 1.0 mL of trifluoroacetic acid R and humidified incubator using 6 ± 1 per cent C0 2 • Add 25 µL of
dilute to 1000 mL with water for chromatography R; a 5.0 g/L sterile solution of tetrazolium bromide R to each well.
- mobile phase B: to 100 mL of water for chromatography R Reincubate for 5 h. Remove the plates from the incubator and
add 1.0 mL of trifluoroacetic acid R and 900 mL of add to each well 100 µL of a 240 g/L solution of sodium dodecyl
acetonitrile Rl ; sulfate R previously adjusted to pH 2. 7 with hydrochloric acid.
Time Mobile phase A Mobile phase B Reincubate overnight.
(min) (per cent V/V) (per cent V/V) Determine the relative quantity of purple formazan product
o - 30 64-? 44 36-? 56 formed in each well by measuring the absorbance (2.2.25)
using a 96-well microtitre plate reader. Read each plate at
30 - 35 44 -7 o 56 -7 100
570 nm and at 690 nm. Subtract the reading at 690 nm from
35 - 45 o 100 the reading at 570 nm. Analyse the data by fitting a sigmoidal
<lose-response curve to the data obtained and by using a
45 - 50 o-? 64 100 -7 36
suitable statistical method, for example the 4-parameter model
50 - 60 64 36 (see 5.3).
The estimated potency is not less than 80 per cent and not
Flow rate: 1.2 mL/min. more than 125 per cent of the stated potency. The confidence
Detection: spectrophotometer at 214 nm. limits (P = 0.95) of the estimated potency are not less than
Injection: 100 µL of test solution (a), reference solutions (a) 74 per cent and not more than 136 per cent of the stated
and (b). potency.
System suitability: reference solution (b) : STORAGE
• ••
- rerenrzon • •
rime: 1
mo1gramosun .. •
= aouuL
- 1_ - - ... ""'
I"\ ~ --
L.L. 111111,
-- -

- repeatability: maximum relative standard deviation of below - 65 ºC.
5.0 per cent after 4 injections,
- resolution: mínimum 2 between the peaks due to albumin LABELLING
and molgramostim. The label states:
Limits: - the content, in milligrams of protein per millilitre,
- any impurity: for each impurity, maximum 1.5 per cent, - the potency, in International Units per milligram of protein.
- total of impurities eluting between 5 min and 30 min:
maximum 4 per cent.
Bacteria! endotoxins (2. 6.14): less than 5 IU in the volume 07/2018:1449
that contains 1.0 mg of protein.
ASSAY
Protein. Liquid chromatography (2.2.29) as described in the
test for related proteins. MOMETASONE FUROATE
Injection: 150 µL of test solution (b) and reference solution (b).
Calculate the content of molgramostim using the declared Mometasoni furoas
content of molgramostim in molgramostim CRS.
Potency. Determination of the biological activity of
molgramostim concentrated solution based on the stimulation
of proliferation of TF-1 cells by molgramostim.
The following method uses the conversion of tetrazolium
bromide (MTT) as a staining method. Validated alternative
stains such as Almar blue have also been found suitable.
o
TF-1 cells are incubated with varying dilutions of test
and reference preparations of molgramostim. They are C27H30Cl206 Mr 521.4
then incubated with a solution of MTT. This cytochemical [83919-23-7]
stain is converted by cellular dehydrogenases to a purple
DEFINITION
formazan product. The formazan is then measured
spectrophotometrically. The potency of the preparation to 9,21-Dichloro-11 ~-hydroxy-16a-methyl-3,20-dioxopregna
be examined is determined by comparison of the dilutions l,4-dien-17-yl furan-2-carboxylate.
of the test preparation with the dilutions of the appropriate Content: 98.0 per cent to 102.0 per cent (dried substance).
Mometasone furoate EUROPEAN PHARMACOPOEIA 9.5
CHARACTERS Related substances. Liquid chromatography (2.2.29). Prepare

Appearance: white or almost white powder. the solutions immediately before use.
Solubility: practically insoluble in water, soluble in acetone and Solvent mixture. Mix 200 mL of acetonitrile R and 200 mL of
-
in methylene chloride, slightly soluble in ethanol (96 per cent). water R, then add 0.4 mL of acetic acid R.
IDENTIFICATION
First identification: A, E, F.
Test solution (a). Dissolve 2S.O mg of the substance to be
examined in lS mL of acetonitrile R and dilute to SO.O mL
Test solution (b ). Dilute S.O mL of test solution (a) to 2S.O mL
Second identification: B, C, D.
-A. Infrared absorption spectrophotometry (2.2.24).
Comparison: mometasone furoate CRS.

B. Thin-layer chromatography (2.2.27).
Test solution. Dissolve 10 mg of the substance to be
Reference solution (a). Dissolve S mg of mometasone furoate
for system suitability CRS (containing impurities C and J) in
3 mL of acetonitrile R and dilute to 10.0 mL with the solvent
mixture.
Reference solution (b). Dilute 1.0 mL of test solution (a) to
examined in methylene chloride R and dilute to 1O mL with 100.0 mL with the solvent mixture. Dilute 1.0 mL of this
the same solvent.
Reference solution (a). Dissolve 20 mg of mometasone Reference solution (c). Dissolve 2S.O mg of mometasone
furoate CRS in methylene chloride R and dilute to 20 mL furoate monohydrate CRS in lS mL of acetonitrile R and dilute
to SO.O mL with the solvent mixture. Dilute S.O mL of the
Reference solution (b ). Dissolve 1O mg of anhydrous solution to 2S.O mL with the solvent mixture.
beclometasone dipropionate CRS in reference solution (a) Column:
and dilute to 10 mL with reference solution (a).
- size: l = 0.2S m, 0 = 4.6 mm;
Plate: TLC silica gel F254 plate R.
- stationary phase: end-capped octadecylsilyl silica gel for
Mobile phase: add a mixture of 1.2 volumes of water R and chromatography R (S µm).
8 volumes of methanol R to a mixture of lS volumes of
ether R and 77 volumes of methylene chloride R. Mobile phase: acetonitrile R, water for chromatography R
(SO:SO V/V).
Application: S µL.
Development: overa path of lS cm.
Detection: spectrophotometer at 2S4 nm.
Drying: in air.
Injection: 20 µL of test solution (a) and reference solutions (a)
Detection A: examine in ultraviolet light at 2S4 nm.
and (b).
Results A: the principal spot in the chromatogram obtained
Run time: 3.5 times the retention time of mometasone furo ate.
with the test solution is similar in position and size to
the principal spot in the chromatogram obtained with Identification of impurities: use the chromatogram supplied
reference solution (a). with mometasone furoate for system suitability CRS and the
chromatogram obtained with reference solution (a) to identify
Detection B: spray with alcoholic solution of sulfuric acid R.
the peaks due to impurities e and J.
Heat at 120 ºC for 10 min or until the spots appear. Allow to
cool; examine in daylight and in ultraviolet light at 36S nm. Relative retention with reference to mometasone furoate
Results B: the principal spot in the chromatogram obtained (retention time = about 24 min): impurity e = about 0.9;
impurity J = about l .S.
with the test solution is similar in position, colour in
daylight, fluorescence in ultraviolet light at 36S nm and System suitability: reference solution (a):
size to the principal spot in the chromatogram obtained - resolution: minimum 2.5 between the peaks due to
with reference solution (a). impurity e and mometasone furoate.
System suitability: reference solution (b): Limits:
- the chromatogram shows 2 spots which, when examined - impurity J: not more than 1.S times the area of the principal
in ultraviolet light at 36S nm, may not be completely peak in the chromatogram obtained with reference
separated. solution (b) (O.lS per cent);
C. Add about 2 mg to 2 mL of sulfuric acid R and shake to - unspecified impurities: for each impurity, not more than the
dissolve. Within lS mina light yellow colour develops. area of the principal peak in the chromatogram obtained
When examined in ultraviolet light at 36S nm, no with reference solution (b) (0.10 per cent);
fluorescence is seen. Add this solution to 10 mL of water R - total: not more than 3 times the area of the principal peak
and mix. The colour fades and there is no fluorescence. in the chromatogram obtained with reference solution (b)
D. Mix 80 mg with 0.30 g of anhydrous sodium carbonate R (0.3 per cent);
and ignite in a crucible until an almost white residue is - disregard limit: O.S times the area of the principal peak in
obtained. Allow to cool and dissolve the residue in S mL of the chromatogram obtained with reference solution (b)
dilute nitric acid R; filter. To 1 mL of the filtrate add 1 mL of (O.OS per cent).
water R. The solution gives reaction (a) of chlorides (2.3.1).
Loss on drying (2.2.32): maximum O.S per cent, determined
E. Examine the chromatograms obtained in the assay. on 1.000 g by drying in an oven at lOS ºC.
with test solution (b) is similar in retention time and size ASSAY
to the principal peak in the chromatogram obtained with Liquid chromatography (2.2.29) as described in the test for
reference solution (c). related substances with the following modification.
F. Loss on drying (see Tests). Injection: test solution (b) and reference solution (c).
TESTS Calculate the percentage content of C27 H 30 Cl 20 6 taking
into account the assigned content of mometasone furoate
Specific optical rotation (2.2. 7) : + SO to + SS (dried monohydrate CRS.
substance).
Dissolve SO.O mg in ethanol (96 per cent) R and dilute to IMPURITIES
1O.O mL with the same solvent. Specified impurities: J.

EUROPEAN PHARMACOPOEIA 9.5 Mometasone furoate

use (2034). lt is therefore not necessary to identify these
impurities for demonstration of compliance. See also 5.1 O. o
Control of impurities in substances for pharmaceutical use): A, o
AC~&R~ULKLUM~RQR~I~
F. 9,21-dichloro-l l~-hydroxy-16a-methyl-3,6,20-
trioxopregna-l,4-dien-l 7-yl furan-2-carboxylate,
A. 21-chloro-16a-methyl-3,20-dioxopregna-l,4,9(1 l)-trien- o
l 7-yl furan-2-carboxylate,
G. 9,21-dichloro- l l ~, l 7-dihydroxy- l 6a-methylpregna- l ,4-
diene-3,20-dione (mometasone),
o o
B. 9-chloro- l 7~-(2,2-dioxo-2,5-dihydro- l ,2A6-oxathiol-4-yl)- H. 9-chloro-11 ~,2 l-dihydroxy- l 6a-methyl-3,20-dioxopregna-

l l ~-hydroxy- l 6a- methyl-3-oxoandrosta-1,4-dien- l 7a-yl l ,4-dien- l 7-yl furan-2-carboxylate,
furan-2-carboxylate,
C. 2 l -chloro- l 6a-methyl-3, 11,20-trioxopregna-l ,4-dien-17 -yl

furan-2-carboxylate, l. 6~-acetoxy-9,21-dichloro-l l ~-hydroxy-16a-methyl-3,20-
dioxo-5~ -pregn -l -en-17 -yl furan -2-carboxylate,
D. 21-chloro-9,l l ~-epoxy-16a-methyl-3,20-dioxo-9~-pregna
l,4-dien-l 7-yl furan-2-carboxylate,
J. 9,21-dichloro- l l ~-hydroxy-6a, l 6a-dimethyl-3,20-
dioxopregna- l ,4-dien- l 7-yl furan-2-carboxylate,
E. 9,2 l-dichloro-16a-methyl-3,20-dioxopregna-l ,4-diene- K. 9-chloro-l l~,l 7,21-trihydroxy-16a-methylpregna-l,4-

l l ~, 17 -diyl di( furan- 2-carboxylate), diene-3,20-dione,
Mometasone furoate monohydrate EUROPEAN PHARMACOPOEIA 9.5
o
o
L. 9,l lp-epoxy-l 7,21-dihydroxy-16a-methyl-9p-pregna-1,4-
diene-3,20-dione, R. 9-chloro-l lp-hydroxy-16a-methyl-21-[(methyl-
sulfonyl)oxy]-3,20-dioxopregna- l ,4-dien-17-yl
o furan-2-carboxylate,
O CIO
CH3·---0~
····H 'o_)
o
M. 9-chloro-l lp,l 7-dihydroxy-16a-methylpregna-l,4-diene- o

3,20-dione, S. 9,21-dichloro-1 lp-hydroxy-16p-methyl-3,20-dioxopregna-
l,4-dien-17-yl furan-2-carboxylate,
o
o
T. 9,21-dichloro-llp-hydroxy-16a-methyl-3,20-dioxopregna-
N. 9-chloro-1l~,l7-dihydroxy-16a-methyi-3,20-dioxopregna- l,4-dien-17-yl 5-chlorofuran-2-carboxylate,
1,4-dien-21-yl methanesulfonate, U. unknown structure.
07/2018:2858
MOMETASONE FUROATE
o MONOHYDRATE
O. 9-chloro-11p,l7-dihydroxy-16a-methyl-3,20-dioxopregna-
1,4-dien-21-yl acetate,
Mometasoni furoas monohydricus
o
C27H30Cl206•H20 Mr 539.4
o [141646-00-6]
P. 9-chloro-l lp, 17-dihydroxy-16a-methyl-3,20-dioxopregna- DEFINITION

l,4-dien-21-yl furan-2-carboxylate, 9,21-Dichloro-11 P-hydroxy-16a-methyl-3,20-dioxopregna-
l ,4-dien-17 -yl furan-2-carboxylate monohydrate.
CHARACTERS
Appearance: white or almost white powder.
Solubility: practically insoluble in water, soluble in acetone and
in methylene chloride, slightly soluble in ethanol (96 per cent).
o
IDENTIFICATION
Q. 21-chloro-9,llp-epoxy-17-hydroxy-16a-methyl-9P- A. Infrared absorption spectrophotometry (2.2.24).
pregna-l,4-diene-3,20-dione, Comparison: mometasone furoate monohydrate CRS.

EUROPEAN PHARMACOPOEIA 9.S Mometasone furoate monohydrate
B. Examine the chromatograms obtained in the assay. Injection: test solution (b) and reference solution (c).
Results: the principal peak in the chromatogram obtained Calculate the percentage content of C 27H 30 Cl20 6 taking
with test solution (b) is similar in retention time and size into account the assigned content of mometasone furoate
to the principal peak in the chromatogram obtained with monohydrate CRS.
reference solution (e).
C. Water (see Tests). IMPURITIES
TESTS if present at a sufficient level, be detected by one or other of
Specific optical rotation (2.2.7): + SO to + SS (anhydrous the tests in the monograph. They are limited by the general
substance). acceptance criterion for other/unspecified impurities and/or
Dissolve SO.O mg in ethanol (96 per cent) R and dilute to by the general monograph Substances far pharmaceutical
10.0 mL with the same solvent. use (2034). It is therefore not necessary to identify these
Related substances. Liquid chromatography (2.2.29). Prepare Control of impurities in substances far pharmaceutical use): A,
the solutions immediately befare use. B, C, D, E, F, G, H, I, J, K, L, M, N, O, P, Q, R, S, T, U.
Solvent mixture. Mix 200 mL of acetonitrile R and 200 mL of
water R, then add 0.4 mL of acetic acid R.
Test solution (a). Dissolve 2S.O mg of the substance to be
examined in lS mL of acetonitrile R and dilute to SO.O mL
Test solution (b). Dilute S.O mL of test solution (a) to 2S.O mL
o
Reference solution (a). Dissolve S mg of mometasone furoate
far system suitability CRS (containing impurity C) in 3 mL of A. 21-chloro-16a-methyl-3,20-dioxopregna-l,4,9( 11)-trien-
acetonitrile R and dilute to 10.0 mL with the solvent mixture. 17 -yl furan-2-carboxylate,
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
Reference solution (c). Dissolve 2S.O mg of mometasone
furoate monohydrate CRS in lS mL of acetonitrile R and dilute
to SO.O mL with the solvent mixture. Dilute S.O mL of the
solution to 2S.O mL with the solvent mixture.
Column:
- size: l = 0.2S m, 0 = 4.6 mm; o
- stationary phase: end-capped octadecylsilyl silica gel far
B. 9-chloro-17~-(2,2-dioxo-2,S-dihydro-l,2;\ 6 -oxathiol-4-yl)-
chromatography R (S µm).
11 ~-hydroxy-16a-methyl-3-oxoandrosta-1,4-dien-17a-yl
Mnhifo ohase: acetnnitrile R. water far chromatoCTraDhv R furan-2-carboAylatc,
(so:~so v1V). - _, º i /

Detection: spectrophotometer at 2S4 nm.
Injection: 20 µL of test solution (a) and reference solutions (a)
and (b).
Run time: 3.5 times the retention time of mometasone furoate.
Identification of impurities: use the chromatogram supplied o
with mometasone furoate far system suitability CRS and the
chromatogram obtained with reference solution (a) to identify C. 21-chloro-16a-methyl-3,l l,20-trioxopregna-l,4-dien-17-yl
the peak due to impurity C. furan-2-carboxylate,
Relative retention with reference to mometasone furoate
(retention time= about 24 min): impurity e= about 0.9.
impurity e and mometasone furoate.
Limits:
- unspecified impurities: for each impurity, not more than the o
area of the principal peak in the chromatogram obtained
with reference solution (b) (0.10 per cent); D. 21-chloro-9,11 ~-epoxy-16a-methyl-3,20-dioxo-9~-pregna-
- total: not more than 3 times the area of the principal peak 1,4-dien-17-yl furan-2-carboxylate,
(0.3 per cent);
- disregard limit: O.S times the area of the principal peak in
(O.OS per cent).
Water (2.5.12): 2.S per cent to 4.0 per cent, determined on
0.200 g.
o
ASSAY
Liquid chromatography (2.2.29) as described in the test for E. 9,21-dichloro-16a-methyl-3,20-dioxopregna-1 ,4-diene-
related substances with the following modification. 11~,17-diyl di(furan-2-carboxylate),
General Notices (1) apply to all monographs and other texts S72S
Mometasone furoate monohydrate EUROPEAN PHARMACOPOEIA 9.5
o o
o
L. 9,11 ~-epoxy-17,21-dihydroxy-16a-methyl-9~-pregna-1,4-
F. 9,21-dichloro-11~-hydroxy-16a-methyl-3,6,20- diene-3,20-dione,
trioxopregna-1,4-dien-l 7-yl furan-2-carboxylate,
o
o
o
M. 9-chloro-11~,l 7-dihydroxy-16a-methylpregna-1,4-diene-
G. 9,21-dichloro-11~,17 -dihydroxy-16a-methylpregna-1,4- 3,20-dione,
diene-3,20-dione (mometasone),
o o
H. 9-chloro-11 ~,2 l -dihydroxy-16a.-methyl-3,20-dioxopregna- N. 9-chloro-llp,17-dihydroxy-16a-methyl-3,20-dioxopregna-
l ,4-dien-17-yl furan-2-carboxylate, l,4-dien-21-yl methanesulfonate,
O. 9-chloro-11~,17-dihydroxy-16a-methyl-3,20-dioxopregna-
I. 6~-acetoxy-9,21-dichloro-l lp-hydroxy-16a-methyl-3,20- 1,4-dien-21-yl acetate,
dioxo-5~-pregn-1-en-17-yl furan-2-carboxylate,
o
o
J. 9,21-dichloro-11~-hydroxy-6a,16a-dimethyl-3,20-
P. 9-chloro-11~,17-dihydroxy-16a-methyl-3,20-dioxopregna
dioxopregna-1,4-dien-17 -yl furan-2-carboxylate, l,4-dien-21-yl furan-2-carboxylate,
o o
K. 9-chloro-11~,l 7,21-trihydroxy-16a-methylpregna-1,4- Q. 21-chloro-9,l lp-epoxy-17-hydroxy-16a-methyl-9~

diene-3,20-dione, pregna-l,4-diene-3,20-dione,

EUROPEAN PHARMACOPOEIA 9.5 Mometasone furoate monohydrate
o
o T. 9,21-dichloro-11~-hydroxy-16a-methyl-3,20-dioxopregna
R. 9-chloro-11~-hydroxy-16a-methyl-21-[(methyl
l,4-dien-17 -yl 5-chlorofuran-2-carboxylate,
sulfonyl)oxy]-3,20-dioxopregna- l ,4-dien-17-yl U. unknown structure.
furan-2-carboxylate,
O CIO
CH3----0~
····H 6-J
o
S. 9,21-dichloro-l l~-hydroxy-16~-methyl-3,20-dioxopregna
l,4-dien-l 7-yl furan-2-carboxylate,

N
Nadroparin calcium ............................................................... 5731 Neostigmine metilsulfate ....................................................... 5733

EUROPEAN PHARMACOPOEIA 9.S Nadroparin calcium
01/2008:1134 Test solution (a). To 10.0 mg of the substance to be examined,

corrected 9.5 add 1.0 mL of water R.
Test solution (b). To 10.0 mg of the substance to be examined,
add O.SO mL of water R and O.SO mL of the interna! standard
solution.
Reference solution (a). Dilute 1.0 mL of anhydrous ethanol R
NADROPARIN CALCIUM to 100.0 mL with water R. Dilute 0.5 mL of the solution to
20.0 mL with water R.
N adroparinum calcicum Reference solution (b). To O.SO mL ofreference solution (a),
add O.SO mL of the interna! standard solution.
Column:
- material: nickel;
- size: l = l.S m, 0 = 2 mm;
- stationary phase: ethylvinylbenzene-divinylbenzene
copolymer R (lS0-180 µm).
Carrier gas: helium for chromatography R or nitrogen for
chromatography R.
Flow rate: 30 mL/min.
0
'so3-
Static head-space conditions that may be used:
= =
R H or 80 3 ( 1/ 2 Ca) , R' H or 80 3( 1/2 Ca) or CO-CH 3
- equilibration temperature: 90 ºC;
= = =
R2 H and R3 C0 2 ( 1/ 2 Ca) or R2 C0 2 ( 1/ 2 Ca) and R3 =H - equilibration time: lS min;
- pressurisation time: 1 min.
DEFINITION
Temperature:
Calcium salt of low-molecular-mass heparin obtained
by nitrous acid depolymerisation of heparin from pork - column: lSO ºC;
intestinal mucosa, followed by fractionation to eliminate - injection port and detector: 2SO ºC.
selectively most of the chains with a molecular mass Detection: flame ionisation.
lower than 2000. The majority of the components have Identification of peaks: use the chromatogram obtained with
a 2-0-sulfo-a-L-idopyranosuronic acid structure at the reference solution (b) to identify the peaks due to ethanol
non-reducing end anda 6-0-sulfo-2,S-anhydro-D-mannitol and 2-propanol.
structure at the reducing end of their chain.
Retention time: ethanol = about 2.S min; 2-propanol = about
Nadroparin calcium complies with the monograph 4 min.
Low-molecular-mass heparins (0828) with the modifications
and additional requirements below. Calculate the percentage content m!m of ethanol taking its
density at 20 ºC to be 0.792 g/mL.
The mass-average relative molecular mass ranges between
3600 and SOOO with a characteristic vaiue oÍ about 4300.
Limit:
The degree of sulfatation is about 2 per disaccharide unit.
- ethanol: maximum 1.0 per cent m!m.
The potency is not less than 9S IU and not more than 130 IU N-NO groups: maximum 0.2S ppm.
of anti-factor Xa activity per milligram, calculated with The content of N-NO-groups is determined by cleavage
reference to the dried substance. The ratio of anti-factor Xa of the N-NO bond with hydrobromic acid in ethyl acetate
activity to anti-factor IIa activity is between 2.S and 4.0. under a reflux condenser and detection of the released NO
by chemiluminescence.
IDENTIFICATION Description of the apparatus (Figure 1134.-1). Use a SOO mL
Carry out identification test A as described in the monograph borosilicate glass round-bottomed flask, above which is
Low-molecular-mass heparins (0828) using nadroparin attached a condenser which is equipped with:
calcium CRS. - on one side, a torion joint through which a stream of
Carry out identification test C as described in the monograph argon R can be introduced via a cannula;
Low-molecular-mass heparins (0828). The following - on the other side, a screw joint with a piston equipped with
requirements apply. a septum through which the reference solution and test
The mass-average relative molecular mass ranges between solution will be injected.
3600 and SOOO. The mass percentage of chains lower than The round-bottomed flask is connected in series to 3 bubble
2000 is not more than 1S per cent. The mass percentage of traps which are themselves connected to 2 cold traps, which
chains between 2000 and 8000 ranges between 7S per cent and are in turn connected to a chemiluminescence detector.
9S per cent. The mass percentage of chains between 2000 and Suitable tubing ensures the junctions are leak-free.
4000 ranges between 3S per cent and SS per cent.
Preparation of the chemiluminescence detector. Switch on the
TESTS chemiluminescence detector 48 h before use and start the
vacuum pump. The vacuum must be less than O.S mm Hg. 1 h
Appearance of solution. The solution is not more opalescent before use, open the oxygen valve at a pressure of 0.2 MPa
than reference suspension II (2.2.1) and not more intensely anda flow rate of 9.4 mL/min.
coloured than reference solution Y5 (2.2.2, Method JI).
Preparation of the bubble trap. In each bubble trap, place
Dissolve O.S g in water R and dilute to 10 mL with the same 30 mL of a 300 g/L solution of sodium hydroxide R in water R.
solvent.
Preparation of the cold traps.
Ethanol. Head-space gas chromatography (2.2.28).
- Trap at - 120 ºC: Slowly add liquid nitrogen to an
Interna[ standard solution. Dilute 1.0 mL of 2-propanol R isothermic flask containing 2SO mL of anhydrous ethanol R
to 100.0 mL with water R. Dilute 1.0 mL of the solution to whilst stirring with a wooden spatula until a paste is
SO.O mL with water R. obtained. Place the cold trap in the isothermic flask
Blank solution. l.O mL of water R. prepared as described.
N adroparin calcium EUROPEAN PHARMACOPOEIA 9.5
VALVE
...._
~
~
-15ºC
~
~
~
~
ARGON
NaOH 300 g/L
Liq. N2/ETHANOL Liq. N2/2-METHYLBUTANE
(BUBBLE TRAPS)
-120ºC -160ºC
SEPTUM (COLO TRAPS)
INJECTION
TREATED
ETHYL ACETATE
+ HBr
RECORDER CHEMILUMINESCENCE
DETECTOR
..__
HEATER
VACUUM
4mm Hg
Flask: round-bottom borosilicate glass flask equipped with a central rodavis joint, a torion joint on the left neck anda 15 mm screw joint on the right
neck; septum: silicone material, diameter 14 mm and thickness 3.5 mm.
Condenser: height 21 cm and interna! diameter 3 cm, with a lower rodavis joint andan upper torion joint.
Bubble traps: height 24 cm and interna! diameter 2.5 cm; interna! tubing: length 23 cm and interna! diameter 0.5 cm. Equipped with a centrally positioned
rotulex mounting with torion joints on the inlet and outlet.
Cold traps: height 16.5 cm and interna! diameter 4 cm; interna! tubing: length 14 cm and interna! diameter 1.3 cm. Equipped with torion joints on the
inlet and outlet and placed in an isothermic flask: internal depth 22 cm and internal diameter 8 cm.
Tubing: fluorinated ethylene propylene material, internal diameter 3.2 mm and thickness 0.8 mm.
Figure 1134.-1. - Apparatus used for the assay of N-NO groups
- Trap at - 160 ºC: Slowly add liquid nitrogen to an Evacuate the system by slowly turning the valve of the
isothermic flask containing 2SO mL of 2-methylbutane R chemiluminescence detector. At the same time tighten the
whilst stirring with a wooden spatula until a paste is inlet on the chemiluminescence detector.
obtained. Place the cold trap in the isothermic flask
When the system is equilibrated, the vacuum reaches
prepared as described.
4mmHg.
Drying of the 500 mL borosilicate-glass round-bottomed flask
and condenser. Boil SO mL of ethyl acetate R under reflux The signal of the zero adjuster on the chemiluminescence
for 1 h under argon R without connecting the system to the detector is set to 1Oper cent of the full scale of the recorder.
chemiluminescence detector. Through the septum of the SOO mL borosilicate glass
Test solution. Dry the substance to be examined for 12 h over round-bottomed flask, sequentially inject O.S mL of water R,
diphosphorus pentoxide R at 60 ºC under vacuum. Dissolve 2.0 mL of dilute hydrobromic acid R and then another 2.0 mL
0.10 g of the treated substance to be examined in 1.0 mL of of dilute hydrobromic acid R, making sure that the recorder
treated formamide R. Shake the solution obtained for 30 min. pen has returned to the baseline between each injection.
Reference solution. Dilute 0.1 mL of nitrosodipropylamine Inject SO.O µL of the reference solution, then SO.O µL of the test
solution R in 6.0 mL of anhydrous ethanol R. Dilute 0.1 mL of solution after the recorder pen has returned to the baseline.
the solution obtained in 1.0 mL of treated formamide R. (This
Calculate the content of N-NO groups of the substance to be
solution is equivalent to O.OS ppm of N-NO groups).
examine d.
Place SO mL of treated ethyl acetate R in the dry SOO mL
borosilicate glass round-bottomed flask equipped with a Free sulfates. Liquid chromatography (2.2.29).
septum. Connect the round-bottomed flask to the condenser Test solution. Dissolve 30.0 mg of the substance to be
which has been previously cooled to - 1S ºC for 2 h. examined in water R and dilute to 10.0 mL with the same
Connect the argon R cannula and adjust the flow rate to solven t.
0.1 L/min. Check that the system is leak-free. Only the Reference solution. Dissolve 1.4787 g of anhydrous sodium
connector to the chemiluminescence detector remains open sulfate R in water R and dilute to 1000.0 mL with the same
in order to avoid excess pressure. solvent. Dilute 1.0 mL of the solution to 200.0 mL with
Heat the treated ethyl acetate R to boiling. distilled water R (S ppm of sulfate ions).

EUROPEAN PHARMACOPOEIA 9.S Neostigmine metilsulfate
Column: Resolution (2.2.25): minimum 1.9 for the absorbance ratio.

- size: l =SO mm, 0 = 4.6 mm; Absorbance ratio: A 167 /A 161 = 0.84 to 0.87.
- stationary phase: anion-exchange resin. C. Infrared absorption spectrophotometry (2.2.24).
Chemical neutralisation system: neutralisation Comparison: neostigmine metilsulfate CRS.
micromembrane in line with the mobile phase for D. To SO mg add 0.4 g of potassium hydroxide R and 2 mL of
anion detection; continuously pump in counter-flow with a ethanol (96 per cent) R and heat on a water-bath for 3 min,
2.4S g/L solution of sulfuric acid R, at a flow rate of 4 mL/min. replacing the evaporated ethanol {96 per cent). Cool and
Mobile phase: add 2 mL of water R and 2 mL of diazobenzenesulfonic acid
- mobile phase A: 1.91 g/L solution of disodium tetraborate R; solution Rl. An orange-red colour develops.
- mobile phase B: 0.1 M sodium hydroxide; E. Dissolve 0.1 gin S mL of distilled water R and add 1 mL of
barium chloride solution Rl. No precipitate is formed. Add
(min)
2 mL of hydrochloric acid R and heat in a water-bath for
(per cent V/V) (per cent V/V)
1O min. A fine, white precipitate is formed.
o - 15 100 o
15 - 15.5 100 7 o o7 100 TESTS
15.5 - 25.5 o 100
Solution S. Dissolve 2.S g in distilled water R and dilute to
SO mL with the same solvent.
Flow rate: 1.0 mL/min. Appearance of solution. Solution S is clear (2.2.1) and
Detection: conductivity detector with a sensitivity of 30 µS. colourless (2.2.2, Method 11).
Injection: SO µL. Acidity or alkalinity. To 4.0 mL of solution S add 6.0 mL
Identification of peaks: use the chromatogram obtained with of water R and 0.1 mL of phenolphthalein solution Rl. The
the reference solution to identify the principal peak due to solution is colourless. Add 0.3 mL of 0.01 M sodium hydroxide;
the sulfate ion. the solution becomes red. Add 0.4 mL of 0.01 M hydrochloric
Retention time: sulfate ion= about 7.S min. Change the acid; the solution becomes colourless. Add 0.1 mL of methyl
composition of the mobile phase, if necessary, to obtain the
red solution R; the solution becomes redor yellowish-red.
prescribed retention time. Related substances. Liquid chromatography (2.2.29).
Limit: Test solution. Dissolve SO.O mg of the substance to be
- free sulfates: maximum O.S per cent. examined in the mobile phase and dilute to SO.O mL with the
mobile phase.
07/2018:0626 100.0 mL with the mobile phase. Dilute 1.0 mL of this solution
Reference solution (b ). Dissolve 4 mg of 3-dimethylaminophe-
nol R (impurity B) in SO.O mL of the mobile phase. Dilute
1.0 mL of the solution to 200.0 mL with the mobile phase.
NEOSTIGMINE METILSULFATE Reference solution (e). Dissolve the contents of a vial of
neostigmine impurity A CRS in 1.0 mL of reference solution (b).
Neostigmini metilsulfas Reference solution (d). Mix 1.0 mL of the mobile phase and
1.0 mL ofreference solution (a).
N O
º0
H3C, )l ~ + ,,CH 3
N
H3C, ,,.S03-
0
Column:
- size: l = 0.2S m, 0 = 4.0 mm;
1 I \
CH 3 H3C CH3 - stationary phase: base-deactivated end-capped octylsilyl
C 13 H 22 Np 6 S Mr 334.4
silica gel far chromatography R (S µm);
[Sl-60-S] - tempera tu re: 30 ºC.
Mobile phase: to 710 mL of a 3.6 g/L solution of sodium
DEFINITION dihydrogen phosphate R previously adjusted to pH 3.2 with
3-[ (Dimethylcarbamoyl)oxy]-N,N,N-trimethylanilinium phosphoric acid R, add 4.3 g of sodium dodecyl sulfate R and
methyl sulfate. 290 mL of acetonitrile far chromatography R.
Content: 98.S per cent to 101.0 per cent (dried substance). Flow rate: 1.6 mL/min.
CHARACTERS Detection : spectrophotometer at 220 nm.
Appearance: white or almost white, crystalline powder or Injection: SO µL of the test solution and reference solutions (a),
colourless crystals, hygroscopic. (c) and (d).
Solubility: very soluble in water, freely soluble in ethanol Run time: twice the retention time of neostigmine.
(96 per cent). Identification of impurities: use the chromatogram obtained
with reference solution (c) to identify the peaks due to
IDENTIFICATION
impurities A and B.
First identification: A, C.
Relative retention with reference to neostigmine (retention
Second identification: A, B, D, E. time = about 20 min): impurity B = about 0.56;
A. Melting point (2.2.14): 144 ºC to 149 ºC. impurity A= about 0.61.
B. Ultraviolet and visible absorption spectrophotometry System suitability:
(2.2.25).
- resolution: minimum 1.S between the peaks due to
Test solution. Dissolve SO mg in 0.5 M sulfuric acid and impurities B and A in the chromatogram obtained with
dilute to 100 mL with the same acid. reference solution (c);
Spectral range: 230-3SO nm. - signal-to-noise ratio: minimum 2S for the principal peak in
Absorption maxima: 261 nm and 267 nm. the chromatogram obtained with reference solution (d).
General Notices (1) apply to ali monographs and other texts S733
N eostigmine metilsulfate EUROPEAN PHARMACOPOEIA 9.5
Calculation of percentage contents: IMPURITIES

- correction factor: multiply the peak area of impurity B Specified impurities: B.
by 0.5;
- for each impurity, use the concentration of neostigmine in if present at a sufficient level, be detected by one or other of
reference solution (a). the tests in the monograph. They are limited by the general
Limits: acceptance criterion for other/unspecified impurities and/or
- impurity B: maximum 0.01 per cent; by the general monograph Substances for pharmaceutical use
- unspecified impurities: for each impurity, maximum (2034). It is therefore not necessary to identify these impurities
0.10 per cent; for demonstration of compliance. See also 5.1 O. Control of
- total: maximum 0.2 per cent; impurities in substances for pharmaceutical use): A, C.
- reporting threshold: O.OS per cent, except for impurity B.
Sulfates (2.4.13): maximum 200 ppm, determined on
solution S.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on A. 3-hydroxy-N,N,N-trimethylanilinium,
1.0 g.
ASSAY
Dissolve 0.300 gin 150 mL of water R and add 100 mL of
dilute sodium hydroxide solution R. Distil, collecting the
distillate in 40 mL of a 40 g/L solution of boric acid R until the
total volume in the collecting vessel is about 250 mL. Titrate
B. 3-( dimethylamino )phenol,
the solution in the collecting vessel with 0.1 M hydrochloric
acid, using 0.25 mL of methyl red mixed solution R as indicator.
Carry out a blank test.
1 mL of 0.1 M hydrochloric acid is equivalent to 33.44 mg of
C13H22N206S.
STORAGE
In an airtight container, protected from light.

p
Paraffin, light liquid ............................................................... 5737 Pimobendan for veterinary use ............................................. 5738
Paraffin, liquid ........................................................................ 5737 Polyoxypropylene stearyl ether............................................. 5739

EUROPEAN PHARMACOPOEIA 9.5 Paraffin, liquid
07/2018:0240 tube, for 5 s. Loosen the stopper, immediately place the tube
in a water-bath, avoiding contact of the tube with the bottom
or side of the bath, and heat for 1O min. After 2 min, 4 min,
6 min and 8 min, remove the tube from the bath and shake
as vigorously as possible, in the longitudinal direction of the
PARAFFIN, LIGHT LIQUID tu be for 5 s. At the end of 1O min of heating, remove the tube
from the water-bath and allow to stand for 10 min. Centrifuge
at 2000 g for 5 min. The lower layer is not more intensely
Paraffinum perliquidum coloured (2.2.2, Method I) than a mixture of 0.5 mL of blue
primary solution, 1.5 mL of red primary solution, 3.0 mL
DEFINITION
of yellow primary solution and 2 mL of a 1O g/L solution of
Purified mixture of liquid saturated hydrocarbons obtained hydrochloric acid R.
from petroleum.
Solid paraffins. Dry a suitable quantity of the substance to be
CHARACTERS examined by heating at 100 ºC for 2 h and cool in a desiccator
over sulfuric acid R. Place in a glass tube with an internal
Appearance: colourless, transparent, oily liquid, free from
diameter of about 25 mm, close the tube and immerse in a
fluorescence in daylight.
bath of iced water. After 4 h, the liquid is sufficiently clear for
Solubility: practically insoluble in water, slightly soluble in a black line, 0.5 mm wide, to be easily seen against a white
ethanol (96 per cent), miscible with hydrocarbons. background held vertically behind the tube.
IDENTIFICATION STORAGE
First identification: A, C. Protected from light.
Second identification: B, C.
FUNCTIONALITY-RELATED CHARACTERISTICS
Comparison: Ph. Eur. reference spectrum of liquid paraffin. This section provides information on characteristics that are
recognised as being relevant control parameters for one or
B. In a test tube cautiously boil 1 mL with 1 mL of 0.1 M more functions of the substance when used as an excipient
sodium hydroxide, with continuous shaking, for about 30 s. (see chapter 5.15). Sorne of the characteristics described in
On cooling to room temperature, 2 phases separate. To the the Functionality-related characteristics section may also be
aqueous phase add 0.1 mL of phenolphthalein solution R. present in the mandatory part of the monograph since they
The solution becomes red. also represent mandatory quality criteria. In such cases, a
C. Viscosity (see Tests). cross-reference to the tests described in the mandatory part is
included in the Functionality-related characteristics section.
TESTS Control of the characteristics can con tribute to the quality
Acidity or alkalinity. To 1O mL add 20 mL of boiling water R of a medicinal product by improving the consistency of the
and shake vigorously for 1 min. Separate the aqueous layer and manufacturing process and the performance of the medicinal
filter. To 1O mL of the fil trate, add 0.1 mL of phenolphthalein product during use. Where control methods are cited, they are
solution R. The solution is colourless. Not more than 0.1 mL recognised as being suitable for the purpose, but other methods
uf 0.1 M sudium hydroxide is req uired to change the colour of can also be use d. i-Vherever results f ora particular characteristic
the indicator to pink. are reported, the control method must be indicated.
Relative density (2.2.5): 0.810 to 0.875. The following characteristic may be relevant for light liquid
Viscosity (2.2.9): 25 mPa·s to 80 mPa·s. paraffin used as emollient in ointments, as vehicle in eye
preparations or as lubricant in tablets and capsules.
Polycyclic aromatic hydrocarhons. Use reagents for
ultraviolet spectrophotometry. Viscosity (see Tests).
Introduce 25.0 mL into a 125 mL separating funnel with
unlubricated ground-glass parts (stopper, stopcock). Add
07 /2018:0239
25 mL of hexane R which has been previously shaken twice
with one-fifth its volume of dimethyl sulfoxide R. Mix and add
5.0 mL of dimethyl sulfoxide R. Shake vigorously for 1 min
and allow to stand until 2 clear layers are formed. Transfer the
lower layer to a 2nd separating funnel, add 2 mL of hexane R
and shake the mixture vigorously. Allow to stand until 2 clear PARAFFIN, LIQUID
layers are formed. Separate the lower layer and measure
its absorbance (2.2.25) between 260 nm and 420 nm, using Paraffinum liquidum
as the compensation liquid the clear lower layer obtained
by vigorously shaking 5.0 mL of dimethyl sulfoxide R with DEFINITION
25 mL of hexane R for 1 min. Prepare a 7.0 mg/L reference Purified mixture of liquid saturated hydrocarbons obtained
solution of naphthalene R in trimethylpentane R and measure from petroleum.
the absorbance of the solution at the absorption maximum
at 275 nm, using trimethylpentane R as the compensation CHARACTERS
liquid. At no wavelength between 260 nm and 420 nm <loes Appearance: colourless, transparent, oily liquid, free from
the absorbance of the test solution exceed one-third that of the fluorescence in daylight.
reference solution at 275 nm.
Solubility: practically insoluble in water, slightly soluble in
Readily carbonisable suhstances. Use a ground-glass- ethanol (96 per cent), miscible with hydrocarbons.
stoppered tube about 125 mm long and 18 mm in internal
diameter, graduated at 5 mL and 1O mL; wash with hot IDENTIFICATION
water R (temperature at least 60 ºC), acetone R, heptane R and First identification: A, C.
finally with aceto ne R, dry at 100-11 O ºC. Cool in a desiccator.
Second identification: B, C.
Introduce 5 mL of the substance to be examined and add 5 mL
of nitrogen-free sulfuric acid Rl. Insert the stopper and shake A. Infrared absorption spectrophotometry (2.2.24).
as vigorously as possible, in the longitudinal direction of the Comparison: Ph. Eur. reference spectrum of liquid paraffin.
Pimobendan for veterinary use EUROPEAN PHARMACOPOEIA 9.5
B. In a test tube cautiously boíl 1 mL with 1 mL of 0.1 M FUNCTIONALITY-RELATED CHARACTERISTICS

sodium hydroxide, with continuous shaking, for about 30 s. This section provides information on characteristics that are
On cooling to room temperature, 2 phases separate. To the recognised as being relevant control parameters for one or
aqueous phase add 0.1 mL of phenolphthalein solution R. more functions of the substance when used as an excipient
The solution becomes red. (see chapter 5.15). Sorne of the characteristics described in
C. Viscosity (see Tests). the Functionality-related characteristics section may also be
TESTS cross-reference to the tests described in the mandatory part is
Acidity or alkalinity. To 10 mL add 20 mL of boiling water R included in the Functionality-related characteristics section.
and shake vigorously for 1 min. Separate the aqueous layer and Control of the characteristics can con tribute to the quality
filter. To 1O mL of the fil trate, add 0.1 mL of phenolphthalein of a medicinal product by improving the consistency of the
solution R. The solution is colourless. Not more than 0.1 mL manufacturing process and the performance of the medicinal
of 0.1 M sodium hydroxide is required to change the colour of product during use. Where control methods are cited, they are
the indicator to pink. recognised as being suitable for the purpose, but other methods
can also be used. Wherever results for a particular characteristic
Relative density (2.2.5): 0.827 to 0.890. are reported, the control method must be indicated.
Viscosity (2.2.9): 110 mPa·s to 230 mPa·s. The following characteristic may be relevant for liquid paraffin
Polycyclic aromatic hydrocarbons. Use reagents for used as emollient in ointments or as lubricant in tablets and
ultraviolet spectrophotometry. capsules.
Viscosity (see Tests).
Introduce 25.0 mL into a 125 mL separating funnel with
unlubricated ground-glass parts (stopper, stopcock). Add
25 mL of hexane R which has been previously shaken twice
with one-fifth its volume of dimethyl sulfoxide R. Mix and add 07/2018:2179
5.0 mL of dimethyl sulfoxide R. Shake vigorously for 1 min
and allow to stand until 2 clear layers are formed. Transfer the
lower layer to a 2ª separating funnel, add 2 mL of hexane R
and shake the mixture vigorously. Allow to stand until 2 clear
layers are formed. Separate the lower layer and measure
its absorbance (2.2.25) between 260 nm and 420 nm, using PIMOBENDAN FOR VETERINARY USE
as the compensation liquid the clear lower layer obtained
by vigorously shaking 5.o mL of dimethyl suljoxide R with Pimobendanum ad usum veterinarium
25 mL of hexane R for 1 min. Prepare a 7.0 mg/L reference
solution of naphthalene R in trimethylpentane R and measure
the absorbance of the solution at the absorption maximum o
at 275 nm, using trimethylpentane Ras the compensation
liquid. At no wavelength between 260 nm and 420 nm does
the absorbance of the test solution exceed one-third that of the
reference solution at 275 nm. and enantiomer
Readily carbonisable substances. Use a ground-glass-
C 19H 18N 4 0 2 Mr 334.4
stoppered tube about 125 mm long and 18 mm in internal
[74150-27-9]
diameter, graduated at 5 mL and 1O mL; wash with hot
water R (temperature at least 60 ºC), acetone R, heptane R and DEFINITION
finally with acetone R, dry at 100-11 O ºC. Cool in a desiccator.
Introduce 5 mL of the substance to be examined and add 5 mL
(5RS)-6-[2-( 4-Methoxyphenyl)-1H-benzimidazol-5-yl]-5-
methyl-4,5-dihydropyridazin-3(2H)-one.
of nitrogen-free sulfuric acid Rl. Insert the stopper and shake
as vigorously as possible, in the longitudinal direction of the Content: 98.0 per cent to 101.0 per cent (anhydrous substance).
tube, for 5 s. Loosen the stopper, immediately place the tube
in a water-bath, avoiding contact of the tube with the bottom CHARACTERS
or side of the bath, and heat for 1O min. After 2 min, 4 min, Appearance: white or slightly yellowish, hygroscopic powder.
6 min and 8 min, remove the tube from the bath and shake Solubility: practically insoluble in water, freely soluble in
as vigorously as possible, in the longitudinal direction of the dimethylformamide, slightly soluble in acetone and in
tube for 5 s. At the end of 1O min of heating, remove the tube methanol.
from the water-bath and allow to stand for 10 min. Centrifuge mp: about 242 ºC.
at 2000 g for 5 min. The lower layer is not more intensely
coloured (2.2.2, Method I) than a mixture of 0.5 mL of blue IDENTIFICATION
primary solution, 1.5 mL of red primary solution, 3.0 mL
Infrared absorption spectrophotometry (2.2.24).
of yellow primary solution and 2 mL of a 1O g/L solution of
hydrochloric acid R. Comparison: pimobendan CRS.
Solid paraffins. Dry a suitable quantity of the substance to be TESTS
examined by heating at 100 ºC for 2 h and cool in a desiccator
over sulfuric acid R. Place in a glass tube with an internal
diameter of about 25 mm, close the tube and immerse in a Test solution. Dissolve 50 mg of the substance to be examined
bath of iced water. After 4 h, the liquid is sufficiently clear for in methanol R and dilute to 10.0 mL with the same solvent.
a black line, 0.5 mm wide, to be easily seen against a white Reference solution (a). Dilute 1.0 mL of the test solution to
background held vertically behind the tube. 100.0 mL with methanol R. Dilute 2.0 mL of this solution to
10.0 mL with methanol R.
STORAGE Reference solution (b ). Dissolve the contents of a vial of
pimobendan for system suitability CRS (impurities A and B) in
Protected from light. 1.0 mL of methanol R.

EUROPEAN PHARMACOPOEIA 9.5 Polyoxypropylene stearyl ether
Column:
- size: l = 0.125 m, 0 = 4.6 mm;
chromatography Rl (5 µm); and enantiomer
- temperature: 45 ºC. A. (3RS)-4-[2-(4-methoxyphenyl)-1H-benzimidazol-5-yl]-3-

Mobile phase: methyl-4-oxobutanoic acid,
- mobile phase A: dissolve 3.0 g of potassium dihydrogen
o
phosphate R in 950 mL of water for chromatography R,
adjust to pH 2.5 with dilute phosphoric acid R and dilute to
1000 mL with water for chromatography R;
- mobile phase B: acetonitrile for chromatography R;
and enantiomer
o-6 85 -7 80 15 -7 20 B. (SRS)-6-[3-amino-4-( 4-methoxybenzamido )phenyl]-5-
methyl-4,5-dihydropyridazin-3(2H)-one.
- 6 - 20
Flow rate: 1 mL/min.

80 -7 20 20 -7 80
07/2018:2602

Injection: 10 µL.
Relative retention with reference to pimobendan POLYOXYPROPYLENESTEARYL
(retention time = about 8.3 min): impurity A = about 1.3; ETHER
impurity B = about 1.4.
System suitability: reference solution (b): Polyoxypropyleni aether stearylicus
impurities A and B. DEFINITION
Mixture of ethers of polyoxypropylene with linear alcohols,
Limits: mainly stearyl alcohol, obtained by the reaction of stearyl
- unspecified impurities: for each impurity, not more than the alcohol with propylene oxide. It may contain sorne free
area of the principal peak in the chromatogram obtained polyoxypropylene and various amounts of free stearyl alcohol.
with reference solution (a) (0.20 per cent); The number of moles of propylene oxide reacted per mole of
- total: not more than the area of the principal peak in the stearyl alcohol is 11 (nominal value). A suitable antioxidant
chromatogram obtained with reference solution (a) (0.2 per may be added.
cent); CHARACTERS
- disregard limit: 0.5 times the area of the principal peak in Appearance: colourless or pale yellow, clear or slightly turbid
the chromatogram obtained with reference solution (a) liquid.
(0.10 per cent).
Solubility: practically insoluble in water, soluble in ethanol
Heavy metals (2.4.8): maximum 10 ppm. (96 per cent), in mineral oils and in 2-propanol, practically
2.0 g complies with test F. Prepare the reference solution using insoluble in propylene glycol and in glycerol.
2 mL of lead standard solution (1O ppm Pb) R. Relative density: about 0.94 at 25 ºC.
Water (2.5.12): maximum 1.0 per cent, determined on 0.500 g. Refractive index: about 1.448 at 25 ºC.
Sulfated ash (2.4.14): maximum 0.1 per cent, determined on IDENTIFICATION
1.0 g. A. Infrared absorption spectrophotometry (2.2.24).
ASSAY Comparison: polyoxypropylene stearyl ether CRS.
B. Hydroxyl value (see Tests).
Dissolve 0.250 g in 5 mL of anhydrous formic acid R. Add
10 mL of acetic anhydride R and 70 mL of anhydrous acetic C. Saponification value (see Tests).
acid R. Titrate with 0.1 M perchloric acid, determining the D. Viscosity (2.2.9): 83 mPa·s to 95 mPa·s.
end-point potentiometrically (2.2.20).
TESTS
of C 19 H 18N 4 0 2 • Acid value (2.5.1): maximum 2.0.
Hydroxyl value (2.5.3, Method A): 60 to 77.
STORAGE Iodine value (2.5.4, Method A): maximum 3.0.
In an airtight container. Peroxide value (2.5.5, Method A): maximum 5.0.
IMPURITIES Saponification value (2.5.6): maximum 3.0.
Other detectable impurities (the following substances would, Propylene oxide. Head-space gas chromatography (2.2.28).
if present at a sufficient level, be detected by one or other of Propylene oxide stock solution. Weigh 45 g of cold macrogol
the tests in the monograph. They are limited by the general 200 Rl and add 1.000 g (m) of propylene oxide R. Mix carefully
acceptance criterion for other/unspecified impurities and/or by swirling to ensure a homogeneous solution. Add more
by the general monograph Substances for pharmaceutical use macrogol 200 Rl until the total weight is SO.O g and mix
(2034). It is therefore not necessary to identify these impurities again. [NOTE: the solution is stable for 1 month if stored at
for demonstration of compliance. See also 5.1 O. Control of - 20 ºC). Allow to reach room temperature. Dilute O.SO g of
impurities in substances for pharmaceutical use): A, B. this solution to 100.0 mL with water R.
Polyoxypropylene stearyl ether EUROPEAN PHARMACOPOEIA 9.5
Propylene oxide standard solution. Dilute 10.0 mL of propylene Calculate the content of propylene oxide in parts per million
oxide stock solution to 100.0 mL with water R. using the following expression:
Propionaldehyde stock solution. Weigh 0.1 g of A1 x m x 5
propionaldehyde R into a volumetric flask and dilute to
100.0 mL with water R.
Test solution (a). Weigh 1.00 g of the substance to be examined
into a 20 mL head-space vial. Add 0.5 mL of cold water R and A 1 area of the peak due to propylene oxide in the
seal the vial immediately with a polytetrafluoroethylene-coated chromatogram obtained with test solution (a);
silicon membrane and an aluminium cap. Mix carefully. A2 area of the peak due to propylene oxide in the
Test solution (b ). Weigh 1.00 g of the substance to be examined chromatogram obtained with test solution (b);
into a 20 mL head-space vial. Add 0.5 mL of cold propylene mass of the substance to be examined in test
oxide standard solution and seal the vial immediately with solution (a), in grams;
a polytetrafluoroethylene-coated silicon membrane and an mass of the substance to be examined in test
aluminium cap. Mix carefully. solution (b ), in grams;
Reference solution. Introduce 50.0 mL of cold propylene m
oxide standard solution into a volumetric flask, add 0.5 mL of mass of propylene oxide used to prepare the
cold propionaldehyde stock solution and dilute to 100.0 mL propylene oxide stock solution, in grams.
with water R. Introduce 0.5 mL of this solution into a 20 mL Limit:
head-space vial.
- propylene oxide: maximum 5 ppm.
Column:
Water (2.5.12): maximum 0.7 per cent, determined on 0.500 g.
- material: fused silica;
- size: l = 60 m, 0 = 0.32 mm; Sulfated ash (2.4.14): maximum 0.3 per cent.
- stationary phase: poly(dimethyl)siloxane R (film thickness Ignite a suitable crucible at 600 ± 25 ºC for 30 min, allow to
5 µm). cool in a desiccator over silica gel or other suitable desiccant
Carrier gas: helium for chromatography R. and weigh. Place 1.0 g of the substance to be examined in the
crucible and weigh. Carefully ignite and char the substance
using a gas burner in a fume cupboard. Carry out the test for
Split ratio: 10: l. sulfated ash (2.4.14) on the residue obtained, starting from
Static head-space conditions that may be used: "Moisten the substance to be examined ...".
- equilibration temperature: 90 ºC;
- equilibration time: 45 min.
STORAGE
Temperature: In an airtight container.
Time Temperature LABELLING
(mio) (ºC)
The label states the number of moles of propylene oxide
Column o- 5 50
reacted per mole of stearyl alcohol (nominal value).
5 - 31 50 -7 180
FUNCTIONALITY-RELATED CHARACTERISTICS
31 - 32.7 180 -7 230
This section provides information on characteristics that are
32.7 - 37.7 230 recognised as being relevant control parameters for one or
Injection port 150 more functions of the substance when used as an excipient
(see chapter 5.15). So me of the characteristics described in
Detector 250 the Functionality-related characteristics section may also be
Injection: a suitable volume, for example 1.0 mL, of the cross-reference to the tests described in the mandatory part is
gaseous phase of test solutions (a) and (b) and of the reference included in the Functionality-related characteristics section.
solution. Control of the characteristics can con tribute to the quality
Identification of peaks: use the chromatogram obtained with of a medicinal product by improving the consistency of the
the reference solution to identify the peaks due to propylene manufacturing process and the performance of the medicinal
oxide and propionaldehyde. product during use. Where control methods are cited, they are
Relative retention with reference to propylene oxide (retention recognised as being suitable for the purpose, but other methods
time = about 10.4 min): propionaldehyde = about 0.96. can also be used. Wherever results for a particular characteristic
System suitability: reference solution: are reported, the control method must be indicated.
- resolution: minimum 1.5 between the peaks due to The following characteristic may be relevant for
propionaldehyde and propylene oxide; polyoxypropylene stearyl ether used as solvent or emollient in
- signal-to-noise ratio: minimum 10 for the peak due to preparations for cutaneous application.
propylene oxide. Viscosity (see Identification).

R
Raltegravir chewable tablets .................................................. 5743 Raltegravir tablets ................................................................... 5744

EUROPEAN PHARMACOPOEIA 9.5 Raltegravir chewable tablets
07/2018:2939 Column:
- size: l = 0.25 m, 0 = 4.6 mm;
RALTEGRAVIR CHEWABLE TABLETS
Mobile phase:
- mobile phase A: mix 20 volumes of acetonitrile for
Raltegraviri compressi masticabiles chromatography R and 80 volumes of a 1.36 g/L solution of
DEFINITION potassium dihydrogen phosphate R previously adjusted to
pH 3.0 with phosphoric acid R;
Raltegravir chewable tablets contain Raltegravir potassium
(2887). - mobile phase B: acetonitrile for chromatography R;
The tablets comply with the monograph Tablets (0478) and Time Mobile phase A Mobile phase B
with the following additional requirements. (min) (per cent V/V) (per cent V/V)
Content: 95.0 per cent to 105.0 per cent of the content of o- 2 100 o
raltegravir (C20 H 21 FN6 0 5 ) stated on the label. 2 - 27 100 ~ 50
IDENTIFICATION Flow rate: 1.0 mL/min.

Carry out either tests A, B or tests B, C. Detection: spectrophotometer at 220 nm.
A. Record the UV spectrum of the principal peak in the Injection: 15 µL of the test solution and reference solutions (b),
chromatograms obtained with the solutions used in (c) and (d).
the assay with a diode array detector in the range of Identification of impurities: use the chromatogram obtained
190-400 nm. with reference solution (d) to identify the peaks dueto
Results: the UV spectrum of the principal peak in impurities C and D; use the chromatogram obtained with
the chromatogram obtained with the test solution is reference solution (c) to identify the peak dueto impurity E.
similar to the UV spectrum of the principal peak in the Relative retention with reference to raltegravir (retention
chromatogram obtained with reference solution (a). time = about 22 min): impurity D = about 0.7;
B. Examine the chromatograms obtained in the assay. impurity C = about 0.8; impurity E= about 0.96.
Results: the principal peak in the chromatogram obtained System suitability: reference solution (c):
with the test solution is similar in retention time and size - resolution: minimum 1.5 between the peaks due to
to the principal peak in the chromatogram obtained with impurity E and raltegravir.
- correction f actors: multiply the peak are as of the following
Preparation: crush a tablet to a powder and homogenise. impurities by the corresponding correction factor:
Comparison: raltegravir potassium CRS. ;.1..1..1..1.yu.1..1.\..]
..... ,...,,r;hr r -
'-J-
h. irnnnrihr
1.1..v,.1..1..1...1.y ...... .1..1."']
n -
.A.-J -
LI. ,•
1.._ .....
Results: the spectrum obtained shows absorption maxima - for each impurity, use the concentration of raltegravir in
at about 1633 cm- 1, 1515 cm- 1, 1188 cm- 1, 810 cm- 1 and reference solution (b).
728 cm- 1, similar to the spectrum obtained with raltegravir Limits:
potassium CRS.
- impurity C: maximum 0.3 per cent;
Other absorption maxima may be present in the spectra.
- impurity D: maximum 0.2 per cent;
TESTS - unspecified impurities: for each impurity, maximum 0.2 per
Related substances. Liquid chromatography (2.2.29). cent;
Solvent mixture: acetonitrile R, water R {30:70 V/V). - total: maximum 0.8 per cent;
Test solution. Place 10 tablets in an appropriate volumetric - reporting threshold: 0.1 per cent.
flask to obtain a concentration of 1 mg/mL of raltegravir and Dissolution (2.9.3, Apparatus 2). The tablets comply with the
make up to volume with the solvent mixture. Stir vigorously test, unless otherwise justified and authorised.
for 1 h. Centrifuge a portion of the solution and dilute 20.0 mL
Dissolution medium: water R, 900 mL.
of the clear supernatant to 200.0 mL with the solvent mixture.
Rotation speed: 50 r/min.
Reference solution (a). Dissolve 22.0 mg of raltegravir
potassium CRS in the solvent mixture and dilute to 200.0 mL Time: 15 min.
with the solvent mixture. Analysis. Liquid chromatography (2.2.29).
Reference solution (b ). Dilute 1.0 mL of the test solution to Test solutions. Solutions from the dissolution test.
100.0 mL with the solvent mixture. Dilute 2.0 mL of this Reference solution. Using sonication if necessary, dissolve a
solution to 10.0 mL with the solvent mixture. suitable quantity of raltegravir potassium CRS in a suitable
Reference solution (e). Dissolve 2 mg of raltegravir quantity of a mixture of 30 volumes of acetonitrile R
impurity E CRS in the solvent mixture and dilute to 100.0 mL and 70 volumes of water R to obtain a concentration of
with the solvent mixture. Dilute 1.0 mL of the solution to raltegravir corresponding to the theoretical concentration
100.0 mL with the test solution. of raltegravir in the test solution, based on the labelled
Reference solution (d). In order to prepare impurities C and D content of the tablets.
in situ, dissolve 20 mg of raltegravir potassium R in a 40 g/L Column:
solution of sodium hydroxide R and dilute to 10 mL with the - size: l = 0.1 m, 0 = 4.6 mm;
same solvent. Stir the solution for 2 h at room temperature.
To 5 mL of the solution add 5 mL of a 103 g/L solution of - stationary phase: end-capped monolithic octadecylsilyl
hydrochloric acid R and dilute to 50 mL with the solvent silica gel for chromatography R;
mixture. - tempera tu re: 40 ºC.
Raltegravir tablets EUROPEAN PHARMACOPOEIA 9.5
Mobile phase: mix 38 volumes of acetonitrile R and

62 volumes of a 1.36 g/L solution of potassium dihydrogen
phosphate R previously adjusted to pH 3.0 with phosphoric
acid R.
Detection: spectrophotometer at 303 nm. D. N-[2-[4-[[(4-fluorophenyl)methyl]carbamoyl]-5-hydroxy-
Injection: 30 µL. l-methyl-6-oxo-1,6-dihydropyrimidin-2-yl]propan-2-
yl] oxamic acid,
Run time: 1 min.
_,,o l"~X"">rl"~
1.5 per cent determined on 6 injections. H,C \-I "HJ00HH V
Calculate the amount of dissolved raltegravir, expressed as a o
percentage of the content of raltegravir (C20 H 21 FN6 0 5 ) stated
on the label, taking into account the assigned content of E. N-benzyl-5-hydroxy-1-methyl-2-[2-[ (5-methyl-1,3,4-
raltegravir potassium CRS and a conversion factor of 0.9210. oxadiazol-2-yl)formamido ]propan-2-yl]-6-oxo-l,6-
dihydropyrimidine-4-carboxamide.
Acceptance criterion:
- Q = 85 per cent after 15 min.
07/2018:2938
ASSAY
Injection: test solution and reference solution (a). RALTEGRAVIR TABLETS
- repeatability: maximum relative standard deviation of Raltegraviri compressi
1.5 per cent determined on 6 injections. DEFINITION
Calculate the percentage content of raltegravir (C20H 21 FN 6 0 5 ) Raltegravir tablets contain Raltegravir potassium (2887).
taking into account the assigned content of raltegravir The tablets comply with the monograph Tablets (0478) and
potassium CRS anda conversion factor of 0.9210. with the folíowing additional requirements.
Content: 95.0 per cent to 105.0 per cent of the content of
IMPURITIES
raltegravir (C20 H 21 FNp 5 ) stated on the label.
Specified impurities: C, D.
IDENTIFICATION
Other detectable impuríties (the following substances would, if
present at a sufficient level, be detected by one or other of the Carry out either tests A, B or tests B, C.
tests in the monograph): A, B, E. A. Record the UV spectrum of the principal peak in the
chromatograms obtained with the solutions used in
the assay with a diode array detector in the range of
190-400 nm.
Results: the UV spectrum of the principal peak in
the chromatogram obtained with the test solution is
similar to the UV spectrum of the principal peak in the
chromatogram obtained with reference solution (a).
B. Examine the chromatograms obtained in the assay.
A. 2-(2-aminopropan-2-yl)-N-[(4-fluorophenyl)methyl]-
5-hydroxy-l-methyl-6-oxo-l,6-dihydropyrimidine-4-
with the test solution is similar in retention time and size
carboxamide,
to the principal peak in the chromatogram obtained with
Preparation: crush a tablet to a powder and homogenise.
Comparison: raltegravir potassium CRS.
Results: the spectrum obtained shows absorption maxima
at about 1633 cm- 1, 1515 cm-1, 1188 cm-1, 810 cm- 1 and
728 cm- 1, similar to the spectrum obtained with raltegravir
B. 2-[2-[ (E)-[ ( dimethylamino )methylidene] amino ]propan-
potassium CRS.
2-yl]-N-[ (4-fluorophenyl)methyl]-5-hydroxy-l-methyl-6-
oxo-1,6-dihydropyrimidine-4-carboxamide, Other absorption maxima may be present in the spectra.
TESTS
Solvent mixture: acetonitrile R, water R (30:70 V!V).
Test solution. Place 10 tablets in an appropriate volumetric
flask to obtain a concentration of 8 mg/mL of raltegravir and
make up to volume with the solvent mixture. Stir vigorously
for 1 h. Centrifuge a portion of the solution and dilute 5.0 mL
C. 2-[2-[2-(2-acetylhydrazin-l-yl)-2-oxoacetamido ]propan- of the clear supernatant to 100.0 mL with the solvent mixture.
2-yl]-N-[ (4-fluorophenyl)methyl]-5-hydroxy- l-methyl-6- Dilute SO.O mL of this solution to 200.0 mL with the solvent
oxo-1,6-dihydropyrimidine-4-carboxamide, mixture.

EUROPEAN PHARMACOPOEIA 9.5 Raltegravir tablets
Reference solution (a). Dissolve 22.0 mg of raltegravir Analysis. Liquid chromatography (2.2.29).
potassium CRS in the solvent mixture and dilute to 200.0 mL Test solutions. Solutions from the dissolution test.
Reference solution. Using sonication, dissolve a suitable
Referen ce solution (b). Dilute 1.0 mL of the test solution to quantity of raltegravir potassium CRS in a suitable
100.0 mL with the solvent mixture. Dilute 2.0 mL of this quantity of a mixture of 30 volumes of acetonitrile R
solution to 10.0 mL with the solvent mixture. and 70 volumes of water R to obtain a concentration of
Reference solution (e). Dissolve 2 mg of raltegravir raltegravir corresponding to the theoretical concentration
impurity E CRS in the solvent mixture and dilute to 100.0 mL of raltegravir in the test solution, based on the labelled
with the solvent mixture. Dilute 1.0 mL of the solution to content of the tablets.
100.0 mL with the test solution. Column:
Reference solution (d). In order to prepare impurities C and D - size: l = 0.1 m, 0 = 4.6 mm;
in situ, dissolve 20 mg of raltegravir potassium R in a 40 g/L
- stationary phase: end-capped monolithic octadecylsilyl
solution of sodium hydroxide R and dilute to 10 mL with the
silica gel f or chromatography R;
same solvent. Stir the solution for 2 h at room temperature.
To 5 mL of the solution add 5 mL of a 103 g/L solution of - tempera tu re: 40 ºC.
hydrochloric acid R and dilute to 50 mL with the solvent Mobile phase: mix 38 volumes of acetonitrile R and
mixture. 62 volumes of a 1.36 g/L solution of potassium dihydrogen
Column: phosphate R previously adjusted to pH 3.0 with phosphoric
acid R.
- size: l = 0.25 m, 0 = 4.6 mm;
chromatography R (5 µm); Detection: spectrophotometer at 303 nm.
- temperature: 40 ºC. Injection: 10 µL.
Mobíle phase: Run time: 1 min.
- mobile phase A: mix 20 volumes of acetonitrile for System suitability: reference solution:
chromatography R and 80 volumes of a 1.36 g/L solution of - repeatability: maximum relative standard deviation of
potassium dihydrogen phosphate R previously adjusted to 1.5 per cent determined on 6 injections.
pH 3.0 with phosphoric acid R; Calculate the amount of dissolved raltegravir, expressed as a
- mobile phase B: acetonitrile for chromatography R; percentage of the content of raltegravir (C 20 H 21 FN60 5 ) stated
Time
on the label, taking into account the assigned content of
Mobile phase A Mobile phase B
raltegravir potassium CRS anda conversion factor of 0.9210.
o-2 100 o Acceptance criteria:
- 15-45 per cent after 15 min;
2 - 27 100 ~ 50
- Q = 70 per cent after 60 min.
Flow rate: 1.0 mL/min. ASSAY
Injection: 15 µL of the test solution and reference solutions (b ), related substances with the foliowing modifications.
(c) and (d).
with reference solution (d) to identify the peaks due to System suitability: reference solution (a):
impurities C and D; use the chromatogram obtained with - repeatability: maximum relative standard deviation of
reference solution (c) to identify the peak dueto impurity E. 1.5 per cent determined on 6 injections.
Relative retention with reference to raltegravir (retention Calculate the percentage content of raltegravir (C 20 H 21 FN 60 5 )
time = about 22 min): impurity D = about 0.7; taking into account the assigned content of raltegravir
impurity C = about 0.8; impurity E = about 0.96. potassium CRS anda conversion factor of 0.9210.
System suitability: reference solution (c): IMPURITIES
- resolution: mínimum 1.5 between the peaks due to Specified impurities: C, D.
impurity E and raltegravir.
Other detectable impurities (the following substances would, if
Calculation of percentage contents: present at a sufficient level, be detected by one or other of the
- correction f actors: multiply the peak areas of the following tests in the monograph): A, B, E.
H3C CH 3 O
impurity C = 1.6; impurity D = 1.4;
- for each impurity, use the concentration of raltegravir in H,NxyNr~~
reference solution (b).
Limits:
H3C,. 0oH ~F
o
- impurity D: maximum 0.3 per cent; A. 2-(2-aminopropan-2-yl)-N-[(4-fluorophenyl)methyl]-
- unspecified impurities: for each impurity, maximum 0.2 per 5-hydroxy-l-methyl-6-oxo-l,6-dihydropyrimidine-4-
cent; carboxamide,
Dissolution (2.9.3, Apparatus 2). The tablets comply with
the test, unless otherwise justified and authorised. Use sinker
devices.
Dissolution medium: water R, 900 mL. B. 2-[2-[(E)-[(dimethylamino)methylidene]amino]propan-
Rotation speed: 100 r/min. 2-yl]-N-[ (4-fluorophenyl)methyl]-5-hydroxy-l-methyl-6-
Time: 15 min and 60 min. oxo-1,6-dihydropyrimidine-4-carboxamide,
Raltegravir tablets EUROPEAN PHARMACOPOEIA 9.5
C. 2-[2-[2-(2-acetylhydrazin-1-yl)-2-oxoacetamido ]propan- E. N-benzyl-5-hydroxy-1-methyl-2-[2-[ (5-methyl-1,3,4-

2-yl]-N- [(4-fluorophenyl)methyl)-5-hydroxy-1-methyl-6- oxadiazol-2-yl)formamido ]propan-2-yl]-6-oxo-l ,6-
oxo-l ,6-dihydropyrimidine-4-carboxamide, dihydropyrimidine-4-carboxamide.
O H3C CH3 O
HO,C)l~xrNr~~
H3C ... 0oH ~F
o
D. N-[2-[4-[[(4-fluorophenyl)methyl]carbamoyl)-5-hydroxy-
1-methyl-6-oxo-l,6-dihydropyrimidin-2-yl]propan-2-
yl] oxamic a cid,

s
Sevoflurane .............................................................................. 5749 Somatropin concentrated solution ....................................... 5752
Somatropin .............................................................................. 5750 Somatropin for injection ....................................................... 5754

EUROPEAN PHARMACOPOEIA 9.5 Sevoflurane
07/2018:2269 Carrier gas: helium f or chromatography R.
• SEVOFLURANE
Sevofluranum
Split ratio: 1:20.
Temperature:
Column
Time
(min)
o - 10
Temperature
(ºC)
40
CF 3 10 - 26 40 ~ 200
F~O~CF3 26 - 40 200
Injection port 200

C4H 3Fp Mr 200.1
[28523-86-6] Detector 225
DEFINITION Detection: flame ionisation.

l,l,l,3,3,3-Hexafluoro-2-(fluoromethoxy)propane.
lnjection: 2 µL; rinse the syringe with a solution containing
CHARACTERS ethylene chloride R before injection of the reference solutions;
Appearance: clear, colourless, volatile liquid. rinse the syringe with the substance to be examined before
injection of the test solution.
Solubility: slightly soluble in water, miscible with ethanol
(96 per cent). ldentification of impurities: use the chromatogram supplied
Relative density: about 1.52. with sevoflurane for system suitability CRS and the
chromatogram obtained with reference solution (b) to identify
bp: about 59 ºC. the peaks due to impurities A and B.
It is non-flammable.
Relative retention with reference to sevoflurane (retention
It decomposes in the presence of Lewis acids; this time = about 6.6 min): impurity A = about O. 78;
decomposition is inhibited by water in sufficient quantity. impurity B = about 0.83; interna! standard = about 1.35.
IDENTIFICATION System suitability: reference solution (b):
Infrared absorption spectrophotometry (2.2.24). - resolution: minimum 2.0 between the peaks due to
Preparation: examine the substance in the gaseous state or impurities A and B.
in the liquid state. Calculate the relative response factor (F1) for reference
Comparison: sevoflurane CRS. solution (a), using the following expression:
TESTS Mi X R
Acidity or alkalinity. Introduce 20.0 mL of the substance to M2
be examined and 20 mL of carbon dioxide-free water R into a
separating funnel, shake for 3 min and allo'v to stand. Collect M1 mass of the interna! standard in reference
the aqueous upper layer and add 0.2 mL of bromocresol purple solution (a), in milligrams;
solution R. Not more than 0.10 mL of 0.01 M sodium hydroxide mass of the substance to be examined in reference
or 0.60 mL of 0.01 M hydrochloric acid is required to change solution (a), in milligrams;
the colour of the indicator.
R ratio of the area of the peak due to sevoflurane to
Refractive index (2.2.6): 1.2745 to 1.2760. the area of the peak due to the interna! standard
Related substances. Gas chromatography (2.2.28). from the chromatogram obtained with reference
Interna[ standard: methylal R. solution (a).
Test solution. Introduce 20.0 mL of the substance to be Calculate the content of each impurity in the substance to be
examined into a vial and seal with a cap and septum. Using examined, in parts per million, using the following expression:
a microsyringe, add 5 µL of the interna! standard and mix 0.859 X R1 X 250
thoroughly.
Reference solution (a). Introduce 2.0 mL of ethylene chloride R
into a screw-cap vial and immediately seal with a cap and
septum. Using a microsyringe, add about 20 µL of the 0.859 = relative density of the internal standard;
substance to be examined. Record the quantity added, in 1.52 relative density of sevoflurane;
milligrams, of the substance to be examined (M2). Then, using
a microsyringe, add about 20 µL of the interna! standard. R1 ratio of the area of the peak due to the impurity to
Record the quantity added, in milligrams, of the interna! the area of the peak due to the interna! standard
standard (M1). from the chromatogram obtained with the test
solution;
Reference solution (b): sevoflurane for system suitability CRS
(containing impurities A and B). relative response factor for reference solution (a).
Reference solution (e). Introduce 20.0 mL of ethylene Limits:
chloride R into a vial and seal with a cap and septum. Using a
- impurity A: maximum 25 ppm;
microsyringe, add 20 µL of the substance to be examined and
mix thoroughly. Dilute 0.5 mL of this solution to 100.0 mL - impurity B: maximum 100 ppm;
with ethylene chloride R. - unspecified impurities: for each impurity, maximum
Column: 100 ppm;
- material: fused silica; - total: maximum 300 ppm;
- size: l = 30 m, 0 = 0.32 mm; disregard limit: the area of the peak due to sevoflurane in
- stationary phase: poly[(cyanopropyl)(phenyl)] [dimeth- the chromatogram obtained with reference solution (c)
yl]siloxane R (film thickness 3 µm). (5 ppm).
Somatropin EUROPEAN PHARMACOPOEIA 9.5
Fluorides: maximum 2 µg/mL. CF 3
Potentiometry (2.2.36, Method I). Use plastic utensils HO~CF3

throughout this test.
Buffer solution. Dissolve O.S g of sodium citrate R and SS g of C. l,l,l,3,3,3-hexafluoropropan-2-ol.
sodium chloride R in 3SO mL of water R. Carefully add 7S g
of sodium hydroxide R and shake to dissolve. Cool to room 01/2008:0951
temperature and carefully add 22S mL of glacial acetic acid R corrected 9.5
while stirring. Cool and add 300 mL of 2-propanol R. Dilute
with water R to 1000.0 mL. The apparent pH of this solution
is between S.O and S.S.
Test solution. Introduce SO.O mL of the substance to be
examined and SO.O mL of water R into a separating funnel, SOMATROPIN
shake vigorously for 3 min and allow the layers to separate
completely. Dilute 2S.O mL of the upper aqueous layer to Somatropinum
SO.O mL with the buffer solution.
FPTIPLSRLF DNAMLRAHRL HQLAFDTYQE FEEAYIPKEQ
Fluoride standard solution (1000 ppm F). Dissolve 221.0 mg
KYSFLQNPQT SLCFSESIPT PSNREETQQK SNLELLRISL
of sodium fluoride R, previously dried at l SO ºC for 4 h, in
LVYGASDSNV YDLLKDLEEG
water R. Add 1.0 mL of 0.01 M sodium hydroxide and dilute to
QTYSKFDTNS HNDDALLKNY
100.0 mL with water R. ,------,
GLLYCFRKDM DKVETFLRIV QCRSVEGSCG
Reference stock solutions. Dilute the fluoride standard solution
( 1000 ppm F) with water R to obtain solutions having known C990H 152sN262 O 300S7 Mr 22 12S
concentrations of about S µg, 2 µg, 0.5 µg, and 0.2 µg of
fluoride per millilitre. DEFINITION
Reference solutions. Dilute 2S.O mL of each reference stock Protein having the structure (191 amino-acid residues) of
solution to SO.O mL with the buffer solution. the major component of growth hormone produced by the
human pituitary.
Indicator electrode: fluoride-selective.
Content: 91.0 per cent to lOS.O per cent (anhydrous substance).
Reference electrode: silver-silver chloride.
By convention, for the purpose of labelling somatropin
Apparatus: voltmeter capable of a minimum reproducibility preparations, 1 mg of anhydrous somatropin
of ± 0.2 mV. (C990 H 1528N26iÜ 300S7) is equivalent to 3.0 IU of biological
Carry out the measurements on the reference solutions activity.
and test solution. To take measurements, transfer the
solution under test to a 100 mL beaker containing a PRODUCTION
polytetrafluoroethylene-coated magnetic stirring bar, and Somatropin is produced by a method based on recombinant
immerse the electrodes. Allow to stir on a magnetic stirrer DNA (rDNA) technology. During the course of product
with an insulated top until equilibrium is attained (about development, it must be demonstrated that the manufacturing
2-3 min), and record the potential. Rinse the electrodes with process produces a product having a biological activity of
the buffer solution and dry, taking care to avoid damaging the not less than 2.5 IU/mg, using a validated bioassay based on
crystal of the specific-ion electrode. growth promotion and approved by the competent authority.
Calculate the concentration of fluorides using the calibration Somatropin complies with the following additional
curve. requirements.
Non-volatile residue: maximum 100 mg/L. Host-cell-derived proteins. The limit is approved by the
Transfer 10.0 mL to a tared evaporating dish, evaporate to competent authority.
dryness on a water-bath and dry the residue at lOS ºC for 2 h. Host-cell- and vector-derived DNA. The limit is approved
The residue weighs a maximum of 1.0 mg. by the competent authority.
Water (2.5.12): maximum O.OSO per cent m/m, determined CHARACTERS
on 10.0 mL.
Appearance: white or almost white powder.
STORAGE
IDENTIFICATION
In an airtight, stainless-steel container, protected from light.
A. Capillary electrophoresis (2.2.47) as described in the test
IMPURITIES for charged variants with the following modifications.
Specified impurities: A, B. Injection: test solution (b); under pressure or vacuum,
Other detectable impurities (the following substances would, using the following sequence: sample injection for at
if present at a sufficient level, be detected by one or other of least 3 s then CZE buffer injection for 1 s.
the tests in the monograph. They are limited by the general Results: in the electropherogram obtained, only 1 principal
acceptance criterion for other/unspecified impurities and/or peak, corresponding to somatropin, is detected: no
by the general monograph Substances for pharmaceutical doubling of this peak is observed.
use (2034). It is therefore not necessary to identify these B. Examine the chromatograms obtained in the test for related
impurities for demonstration of compliance. See also 5.10. proteins.
Control of impurities in substances for pharmaceutical use): C. Results: the principal peak in the chromatogram obtained
CF 3 with the test solution is similar in retention time and size
F~OACF2 to the principal peak in the chromatogram obtained with
the reference solution.
C. Peptide mapping (2.2.55).
A. 1,1,3,3,3-pentafluoro-2-(fluoromethoxy)prop-1-ene,
CF 3
Test solution. Prepare a solution of the substance to be
H3CO
~ CF3 examined in O.OS M tris-hydrochloride buffer solution
pH 7.5 R to obtain a solution containing 2.0 mg/mL of
B. l,l,l,3,3,3-hexafluoro-2-methoxypropane, somatropin and transfer about 1.0 mL to a tube made
S7SO See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 9.5 Somatropin
from a suitable material such as polypropylene. Prepare Column:

a 1 mg/mL solution of trypsin for peptide mapping R in - size: l = 0.25 m, 0 = 4.6 mm;
O.OS M tris-hydrochloride buffer solution pH 7.S R and add - stationary phase: end-capped butylsilyl silica gel for
30 µL to the solution of the substance to be examined. Cap
chromatography R (5 µm) with a pore size of 30 nm; a
the tube and place in a water-bath at 37 ºC for 4 h. Remove silica saturation column is placed between the pump and
from the water-bath and stop the reaction immediately, the injector valve;
for example by freezing. If analysed immediately using an
automatic injector, maintain at 2-8 ºC. - temperature: 45 ºC.
Mobile phase: propano[ R, O.OS M tris-hydrochloride buffer
Reference solution. Prepare at the same time and in the same solution pH 7.S R (29:71 V/V).
manner as for the test solution, but using somatropin CRS
instead of the substance to be examined. Flow rate: 0.5 mL/min.
(2.2.29). Preconditioning of the column: rinse with 200-500 mL of a
0.1 per cent V/V solution of trifluoroacetic acid R in a 50 per
Column: cent V/V solution of acetonitrile R; repeat as necessary, to
- size: l = 0.25 m, 0 = 4.6 mm; improve column performance.
Injection: 20 µL.
- stationary phase: octylsilyl silica gel f or chromatography R
(5-10 µm) with a pore size of 30 nm; Relative retention with reference to somatropin (retention
time= about 33 min; if necessary adjust the concentration of
- tempera tu re: 30 ºC. propano[ R in the mobile phase) : desamidosomatropin = about
Mobile phase: 0.85.
- mobile phase A: dilute 1 mL of trifluoroacetic acid R to System suitability: resolution solution:
1000 mL with water for chromatography R; - resolution: minimum 1.0 between the peaks due to
desamidosomatropin and somatropin;
- mobile phase B: to 100 mL of water for chromatography R,
- symmetry factor: 0.9 to 1.8 for the peak dueto somatropin.
with acetonitrile Rl; Limit:
- total: maximum 6.0 per cent.
Dimer and related substances of higher molecular mass.
Size-exclusion chromatography (2.2.30): use the normalisation
o - 20 100 -7 80 o -7 20 procedure.
20 - 40 80 -7 75 20 -7 25 Test solution. Prepare a solution of the substance to be
40 - 65 75 -7 50 25 -7 50 examined in 0.02S M phosphate buffer solution pH 7.0 R,
containing 1.0 mg/mL of somatropin.
65 - 70 50 -7 20 50 -7 80
Reference solution. Dissolve the contents of a vial of
somatropin CRS in 0.02S M phosphate buffer solution pH 7.0 R
Flow rate: 1 mL/min. and dilute with the same solution to obtain a concentration
Detection: spectrophotometer at 214 nm. of l.O mgimL.
Injection: 100 µL. Resolution solution. Place 1 vial of somatropin CRS in an oven
at 50 ºC for a period sufficient to generate 1-2 per cent of
System suitability: the chromatograms obtained with the dimer (typically 12-24 h). Dissolve its contents in 0.02S M
test solution and the reference solution are similar to phosphate buffer solution pH 7.0 R and dilute with the same
the chromatogram of somatropin digest supplied with solution to obtain a concentration of 1.0 mg/mL.
somatropin CRS. Column:
Results: the profile of the chromatogram obtained with - size: l = 0.30 m, 0 = 7.8 mm;
the test solution corresponds to that of the chromatogram - stationary phase: hydrophilic silica gel for chromatography R
obtained with the reference solution. of a grade suitable for fractionation of globular proteins in
D. Examine the chromatograms obtained in the assay. the relative molecular mass range of 5000 to 150 000.
Results: the principal peak in the chromatogram obtained Mobile phase: 2-propanol R, 0.063 M phosphate buffer solution
with the test solution is similar in retention time and size pH 7.0 R (3:97 V/V); filter and <legas.
to the principal peak in the chromatogram obtained with Flow rate: 0.6 mL/min.
the reference solution. Detection: spectrophotometer at 214 nm.
Injection: 20 µL.
TESTS Relative retention with reference to somatropin monomer
Related proteins. Liquid chromatography (2.2.29): use the (retention time = 12 min to 17 min): related substances
normalisation procedure. Maintain the solutions at 2-8 ºC and of higher molecular mass = about 0.65; somatropin
use within 24 h. If an automatic injector is used, maintain it dimer = about 0.9.
at 2-8 ºC. System suitability: resolution solution:
Test solution. Prepare a solution of the substance to be - peak-to-valley ratio: minimum 2.5, where HP = height above
examined in O.OS M tris-hydrochloride buffer solution pH 7.S R, the baseline of the peak due to the dimer and Hv = height
containing 2.0 mg/mL of somatropin. above the baseline of the lowest point of the curve
Reference solution. Prepare a solution of somatropin CRS in
O.OS M tris-hydrochloride buffer solution pH 7.S R, containing Limit:
2.0 mg/mL of somatropin. - sum of the peaks with retention times less than that of the
principal peak: maximum 4.0 per cent.
Resolution solution. Dissolve the contents of a vial of
somatropin!desamidosomatropin resolution mixture CRS in Charged variants. Capillary electrophoresis (2.2.47).
O.OS M tris-hydrochloride buffer solution pH 7.S R to obtain a Test solution (a). Prepare a solution of the substance to be
concentration of 2 mg/mL of somatropin. examined containing 1 mg/mL of somatropin.
Somatropin concentrated solution EUROPEAN PHARMACOPOEIA 9.5
Test solution (b). Mix equal volumes of test solution (a) and 01/2008:0950
the reference solution. corrected 9.5
somatropin CRS in water R and dilute with the same solvent
to obtain a concentration of 1 mg/mL.
Capillary: SOMATROPIN CONCENTRATED
- material: uncoated fused silica; SOLUTION
- size: effective length = at least 70 cm, 0 = 50 µm.
Somatropini solutio concentrata
Temperature: 30 ºC.
FPTIPLSRLF DNAMLRAHRL HQLAFDTYQE FEEAYIPKEQ
CZE buffer: 13.2 g/L solution of ammonium phosphate R
KYSFLQNPQT SLCFSESIPT PSNREETQQK SNLELLRISL
adjusted to pH 6.0 with phosphoric acid R and filtered.
Detection: spectrophotometer at 200 nm. QTYSKFDTNS HNDDALLKNY
,-------¡
GLLYCFRKDM DKVETFLRIV QCRSVEGSCG
Set the autosampler to store the samples at 4 ºC during analysis.
C990H152sN262 O300S7 Mr 22 125
Preconditioning of the capillary: rinse with 1 M sodium
hydroxide for 20 min, with water R for 1O min and with DEFINITION
the CZE buffer for 20 min. Solution containing a protein having the structure
Between-run rinsing: rinse with 0.1 M sodium hydroxide for (191 amino-acid residues) of the major component of growth
2 min and with the CZE buffer for 6 min. hormone produced by the human pituitary. It may contain
buffer salts and other auxiliary substances.
Note: rinsing times may be adapted according to the length of
the capillary and the equipment used. Content: 91.0 per cent to 105.0 per cent of the amount of
somatropin stated on the label.
Injection: test solution (a) and the reference solution; under By convention, for the purpose of labelling somatropin
pressure or vacuum, using the following sequence: sample preparations, 1 mg of anhydrous somatropin
injection for at least 3 s then CZE buffer injection for 1 s. (C 990H 1528N 262 Ü 300S7) is equivalent to 3.0 IU of biological
The injection time and pressure may be adapted in order to activity.
meet the system suitability criteria. PRODUCTION
Migration: apply a field strength of 217 V/cm (20 kV for Somatropin concentrated solution is produced by a method
capillaries of 92 cm total length) for 80 min, using the CZE based on recombinant DNA (rDNA) technology. During the
buffer as the electrolyte in both buffer reservoirs. course of product development, it must be demonstrated
Relative migration with reference to somatropin: deamidated that the manufacturing process produces a product having
forms = 1.02 to 1.11. a biological activity of at least 2.5 IU/mg, using a validated
bioassay based on growth promotion and approved by the
System suitability: reference solution: competent authority.
- the electropherogram obtained is similar to the Somatropin concentrated solution complies with the following
electropherogram of somatropin supplied with additional requirements.
somatropin CRS; 2 peaks (Ip 12 ) eluting prior to the Host-cell-derived proteins. The limit is approved by the
principal peak and at least 2 peaks (1 3, 14 ) eluting after the competent authority.
principal peak are clearly visible. Host-cell- and vector-derived DNA. The limit is approved
Note: peak l 2 corresponds to the cleaved form and peak l 4 by the competent authority.
corresponds to the deamidated forms, eluting as a doublet.
CHARACTERS
Limits: Appearance: clear or slightly turbid, colourless solution.
- deamidatedforms: maximum 5.0 per cent;
IDENTIFICATION
- any other impurity: for each impurity, maximum 2.0 per A. Capillary electrophoresis (2.2.47) as described in the test
cent; for charged variants with the following modifications.
- total: maximum 10.0 per cent. Injection: test solution (b); under pressure or vacuum,
using the following sequence: sample injection for at
Water (2.5.32): maximum 10.0 per cent. least 3 s then CZE buffer injection for 1 s.
Bacterial endotoxins (2. 6.14): less than 5 IU /mg, if intended Results: in the electropherogram obtained, only 1 principal
for use in the manufacture of parenteral preparations without peak, corresponding to somatropin, is detected: no
a further appropriate procedure for removal of bacteria! doubling of this peak is observed.
endotoxins. B. Examine the chromatograms obtained in the test for related
proteins.
ASSAY Results: the principal peak in the chromatogram obtained
Size-exclusion chromatography (2.2.30) as described in the with the test solution is similar in retention time and size
test for dimer and related substances ofhigher molecular mass. to the principal peak in the chromatogram obtained with
the reference solution.
Calculate the content of somatropin (C 990 H 1528N 262 Ü 300S7) from C. Peptide mapping (2.2.55).
the declared content of C990 H 1528 N262 0 300S7 in somatropin CRS.
Test solution. Dilute the solution to be examined with
STORAGE
0.05 M tris-hydrochloride buffer solution pH 7.5 R so that
In an airtight container, at a temperature of 2 ºC to 8 ºC. If the it contains 2.0 mg/mL of somatropin and transfer about
substance is sterile, store in a sterile, airtight, tamper-proof 1.0 mL to a tube made from a suitable material such as
container. polypropylene. Prepare a 1 mg/mL solution of trypsin

EUROPEAN PHARMACOPOEIA 9.5 Somatropin concentrated solution
for peptide mapping R in 0.05 M tris-hydrochloride buffer Column:

solution pH 7.5 R and add 30 µL to the solution of the - size: l = 0.25 m, 0 = 4.6 mm;
substance to be examined. Cap the tube and place in a - stationary phase: end-capped butylsilyl silica gel for
water-bath at 37 ºC for 4 h. Remove from the water-bath chromatography R (5 µm) with a pore size of 30 nm; a
and stop the reaction immediately, for example by freezing. silica saturation column is placed between the pump and
If analysed immediately using an automatic injector, the injector valve;
maintain at 2-8 ºC.
Note: If a 2 mglmL somatropin concentration is not
Mobile phase: propano[ R, 0.05 M tris-hydrochloride buffer
obtainable, a similar digest relationship (micrograms of
solution pH 7.5 R (29:71 V/V).
trypsin per milligram of somatropin) may be used.
Reference solution. Prepare at the same time and in the same
manner as for the test solution, but using somatropin CRS Detection: spectrophotometer at 220 nm.
instead of the substance to be examined. Preconditioning of the column: rinse with 200-500 mL of a
CHROMATOGRAPHIC SEPARATION. Liquid chromatography
(2.2.29). cent V/V solution of acetonitrile R; repeat as necessary, to
improve column performance.
Column: Injection: 20 µL.
- size: l = 0.25 m, 0 = 4.6 mm; Relative retention with reference to somatropin
- stationary phase: octylsilyl silica gel for chromatography R (retention time = about 33 min; if necessary adjust
(5-10 µm) with a pore size of 30 nm; the concentration of propanol R in the mobile phase):
- temperature: 30 ºC. desamidosomatropin = about 0.85.
Mobile phase: System suitability: resolution solution:
mobile phase A: dilute 1 mL of trifluoroacetic acid R to - resolution: minimum 1.0 between the peaks due to
1000 mL with water for chromatography R; desamidosomatropin and somatropin;
mobile phase B: to 100 mL of water for chromatography R,
add 1 mL of trifluoroacetic acid R and dilute to 1000 mL Limit:
with acetonitrile Rl; - total: maximum 6.0 per cent.
Time Mobile phase A Mobile phase B Dimer and related substances of higher molecular mass.
(min) (per cent V/V) (per cent V/V) Size-exclusion chromatography (2.2.30) : use the normalisation
o - 20 100 7 80 o7 20
procedure.
Test solution. Dilute the solution to be examined in 0.025 M
20 - 40 80 7 75 20 7 25 phosphate buffer solution pH 7.0 R, so as to contain 1.0 mg/mL
40 - 65 75 7 50 25 7 50 of somatropin.
50 7 20 50 7 80
65 - 70
somatropin CRS in 0.025 M phosphate buffer solution pH 7.0 R
Flow rate: l mL/min.
and dilute with the same solution to obtain a concentration
of 1.0 mg/mL.
Resolution solution. Place 1 vial of somatropin CRS in an oven
Injection: 100 µL. at 50 ºC for a period sufficient to generate 1-2 per cent of
System suitability: the chromatograms obtained with the dimer (typically 12-24 h). Dissolve its contents in 0.025 M
test solution and the reference solution are similar to phosphate buffer solution pH 7.0 R and dilute with the same
the chromatogram of somatropin digest supplied with solution to obtain a concentration of 1.0 mg/mL.
somatropin CRS. Column:
Results: the profile of the chromatogram obtained with - size: l = 0.30 m, 0 = 7.8 mm;
the test solution corresponds to that of the chromatogram - stationary phase: hydrophilic silica gel for chromatography R
obtained with the reference solution. of a grade suitable for fractionation of globular proteins in
D. Examine the chromatograms obtained in the assay. the relative molecular mass range of 5000 to 150 000.
Results: the principal peak in the chromatogram obtained Mobile phase: 2-propanol R, 0.063 M phosphate buffer solution
with the test solution is similar in retention time and size pH 7.0 R (3:97 V/V); filter and <legas.
to the principal peak in the chromatogram obtained with Flow rate: 0.6 mL/min.
the reference solution. Detection: spectrophotometer at 214 nm.
TESTS Injection: 20 µL.
Relative retention with reference to somatropin monomer
Related proteins. Liquid chromatography (2.2.29) : use the
(retention time = 12 min to 17 min): related substances
normalisation procedure. Maintain the solutions at 2-8 ºC and
of higher molecular mass = about 0.65; somatropin
use within 24 h. If an automatic injector is used, maintain it
dimer = about 0.9.
at 2-8 ºC.
System suitability: resolution solution:
Test solution. Dilute the solution to be examined in 0.05 M
tris-hydrochloride buffer solution pH 7.5 R, so as to contain - peak-to-valley ratio: minimum 2.5, where HP = height above
2.0 mg/mL of somatropin. A weaker solution may be prepared, the baseline of the peak due to the dimer and Hv = height
in which case the injection volume is adjusted accordingly. above the baseline of the lowest point of the curve
Reference solution. Prepare a solution of somatropin CRS in
0.05 M tris-hydrochloride buffer solution pH 7.5 R, containing Limit:
2.0 mg/mL of somatropin. - sum of the peaks with retention times less than that of the
Resolution solution. Dissolve the contents of a vial of
somatropin!desamidosomatropin resolution mixture CRS in Charged variants. Capillary electrophoresis (2.2.47).
0.05 M tris-hydrochloride buffer solution pH 7.5 R to obtain a Test solution (a). Dilute the solution to be examined so as to
concentration of 2 mg/mL of somatropin. obtain a concentration of 1 mg/mL of somatropin.
Somatropin for injection EUROPEAN PHARMACOPOEIA 9.5
Test solution (b). Mix equal volumes of test solution (a) and 01/2008:0952
the reference solution. corrected 9.5
somatropin CRS in water R and dilute with the same solvent
Capillary:
SOMATROPIN FOR INJECTION
- material: uncoated fused silica;
- size: effective length = at least 70 cm, 0 = 50 µm. Somatropinum iniectabile
CZE buffer: 13.2 g/L solution of ammonium phosphate R FPTIPLSRLF DNAMLRAHRL HQLAFDTYQE FEEAYIPKEQ
adjusted to pH 6.0 with phosphoric acid R and filtered. KYSFLQNPQT SLCFSESIPT PSNREETQQK SNLELLRISL
Detection: spectrophotometer at 200 nm. QTYSKFDTNS HNDDALLKNY
,----,
Set the autosampler to store the samples at 4 ºC during analysis. GLLYCFRKDM DKVETFLRIV QCRSVEGSCG F
Preconditioning of the capillary: rinse with 1 M sodium C990H1s2sN262Ü300S7 Mr 22 125

hydroxide for 20 min, with water R for 10 min and with
the CZE buffer for 20 min. DEFINITION
Between-run rinsing: rinse with 0.1 M sodium hydroxide for Freeze-dried, sterile preparation of a protein having the
2 min and with the CZE buffer for 6 min. structure (191 amino-acid residues) of the major component
Note: rinsing times may be adapted according to the length of of growth hormone produced by the human pituitary.
the capillary and the equipment used. Content: 89.0 per cent to 105.0 per cent of the amount of
somatropin stated on the label.
lnjection: test solution (a) and the reference solution; under
pressure or vacuum, using the following sequence: sample By convention, for the purpose of labelling somatropin
injection for at least 3 s then CZE buffer injection for 1 s. The preparations, 1 mg of anhydrous somatropin
injection time and pressure may be adapted in order to meet (C990 H 1528N 262 Ü 300S7 ) is equivalent to 3.0 IU of biological
the system suitability criteria. activity.
Migration: apply a field strength of 217 V/ cm (20 kV for Somatropin for injection complies with the requirements of
capillaries of 92 cm total length) for 80 min, using the CZE the monograph Parenteral preparations (0520).
buffer as the electrolyte in both buffer reservoirs.
PRODUCTION
Relative migration with reference to somatropin: deamidated Somatropin for injection is prepared either from Somatropin
forms = 1.02 to 1.11. (0951) or from Somatropin concentrated solution (0950), or by
System suitability: reference solution: a method based on recombinant DNA (rDNA) technology
- the electropherogram obtained is similar to the in which the injectable preparation is produced without
electropherogram of somatropin supplied with the isolation of an intermediate solid or liquid bulk. In
somatropin CRS; 2 peaks (IP I2) eluting prior to the the latter case, during the course of product development,
principal peak and at least 2 peaks (1 3, I4) eluting after the it must be demonstrated that the manufacturing process
principal peak are clearly visible. produces a product having a biological activity of not less
than 2.5 IV /mg, using a validated bioassay based on growth
Note: peak 12 corresponds to the cleaved form and peak 14 promotion and approved by the competent authority. The
corresponds to the deamidated forms, eluting as a doublet. purified preparation, to which buffers and stabilisers may be
Limits: added, is filte red through a bacteria-retentive filter, aseptically
distributed in sterile containers of glass type I (3.2.1) and
- deamidatedforms: maximum 5.0 per cent;
freeze-dried. The containers are immediately sealed so as to
- any other impurity: for each impurity, maximum 2.0 per exclude microbial contamination and moisture.
cent;
Somatropin for injection complies with the following
- total: maximum 10.0 per cent. additional requirements.
Bacteria! endotoxins (2.6.14): less than 5 IV in the volume Host-cell-derived proteins. The limit is approved by the
that contains 1 mg of somatropin, if intended for use in the competent authority.
manufacture of parenteral preparations without a further
Host-cell- and vector-derived DNA. The limit is approved
appropriate procedure for removal of bacteria! endotoxins.
ASSAY Where somatropin for injectian is prepared from Somatropin
(0951) or fram Somatrapin cancentrated salution (0950),
Size-exclusion chromatography (2.2.30) as described in the campliance with the requirements far host-cell-derived
test for dimer and related substances ofhigher molecular mass. proteins, host-cell- and vector-derived DNA, identification
Calculate the content of somatropin (C 990 H 1528N 262 Ü 300S7 ) from test A, identification test C and charged variants need nat be
the declared content of C 990 H 1528 N 262 Ü 300S7 in somatropin CRS. reconfirmed by the manufacturer during subsequent praduction
af somatropin far injection.
STORAGE
CHARACTERS
In an airtight container at a temperature of - 20 ºC. Avoid
repeated freezing and thawing. If the solution is sterile, store Appearance: white or almost white powder.
in a sterile, airtight, tamper-proof container.
IDENTIFICATION
LABELLING A. Capillary electrophoresis (2.2.47) as described in the test
for charged variants with the following modifications.
The label states:
lnjection: test solution (b); under pressure or vacuum,
- the content of somatropin in milligrams per millilitre; using the following sequence: sample injection for at least
- the name and concentration of any auxiliary substance. 3 s then CZE buffer injection for 1 s.
5754 See the infarmation sectian an general managraphs (caver pages)

EUROPEAN PHARMACOPOEIA 9.5 Somatropin for injection
Results: in the electropherogram obtained, only 1 principal TESTS

peak, corresponding to somatropin, is detected: no Related proteins. Liquid chromatography (2.2.29) : use the
doubling of this peak is observed. normalisation procedure. Maintain the solutions at 2-8 ºC and
B. Examine the chromatograms obtained in the test for related use within 24 h. lf an automatic injector is used, maintain at
proteins. 2-8 ºC.
Results: the principal peak in the chromatogram obtained Test solution. Prepare a solution of the substance to be
with the test solution is similar in retention time and size examined in 0.05 M tris-hydrochloride buffer solution pH 7.5 R,
to the principal peak in the chromatogram obtained with containing 2.0 mg/mL of somatropin.
the reference solution. Reference solution. Prepare a solution of somatropin CRS in
0.05 M tris-hydrochloride buffer solution pH 7.5 R, containing
C. Peptide mapping (2.2.55). 2.0 mg/mL of somatropin.
SELECTIVE CLEA VAGE OF THE PEPTIDE BONDS Resolution solution. Dissolve the contents of a vial of
Test solution. Prepare a solution of the substance to be somatropin!desamidosomatropin resolution mixture CRS in
examined in 0.05 M tris-hydrochloride buffer solution 0.05 M tris-hydrochloride buffer solution pH 7.5 R to obtain a
pH 7.5 R to obtain a solution containing 2.0 mg/mL of concentration of 2 mg/mL of somatropin.
somatropin and transfer about 1.0 mL to a tube made Column:
from a suitable material such as polypropylene. Prepare - size: l = 0.25 m, 0 = 4.6 mm;
a 1 mg/mL solution of trypsin for peptide mapping R in - stationary phase: end-capped butylsilyl silica gel for
0.05 M tris-hydrochloride buffer solution pH 7.5 R and add chromatography R (5 µm) with a pore size of 30 nm; a
30 µL to the solution of the substance to be examined. Cap silica saturation column is placed between the pump and
the tube and place in a water-bath at 37 ºC for 4 h. Remove the injector valve;
from the water-bath and stop the reaction immediately,
for example by freezing. If analysed immediately using an
automatic injector, maintain at 2-8 ºC. Mobile phase: propano! R, 0.05 M tris-hydrochloride buffer
solution pH 7.5 R (29:71 V/V).
Reference solution. Prepare at the same time and in the same
manner as for the test solution, but using somatropin CRS
instead of the substance to be examined. Detection: spectrophotometer at 220 nm.
Preconditioning of the column: rinse with 200-500 mL of a
(2.2.29).
cent V/V solution of acetonitrile R; repeat as necessary, to
Column: improve column performance.
- size: l = 0.25 m, 0 = 4.6 mm; lnjection: 20 µL.
Relative retention with reference to somatropin
- stationary phase: octylsilyl silica gel for chromatography R (retention time = about 3 3 min; if necessary adjust
( 5-1 O µm) with ª pore size of 3o nm; the concentration of propano! R in the mobile phase):
- temperature: 30 ºC. desamidosomatropin = about 0.85.
Mobile phase: System suitability: resolution solution:
desamidosomatropin and somatropin;
1000 mL with water for chromatography R;
- mobile phase B: to 100 mL of water for chromatography R Limit:
with acetonitrile Rl; - total: maximum 13.0 per cent.
Dimer and related substances of higher molecular mass.
Time Mobile phase A Mobile phase B Size-exclusion chromatography (2.2.30): use the normalisation
(min) (per cent V/V) (per cent V/V) procedure.
o - 20 100 -7 80 o -7 20 Test solution. Prepare a solution of the substance to be
20 - 40 80 -7 75 20 -7 25 examined in 0.025 M phosphate buffer solution pH 7.0 R,
contaíníng 1.0 mg/mL of somatropín.
40 - 65 75 -7 50 25 -7 50
65 - 70 50 -7 20 50 -7 80 somatropin CRS in 0.025 M phosphate buffer solution pH 7.0 R
and dilute wíth the same solution to obtain a concentration
Flow rate: 1 mL/min. of 1.0 mg/mL.
Detection: spectrophotometer at 214 nm. Resolution solution. Place 1 vial of somatropin CRS in an oven
at 50 ºC for a period sufficient to generate 1-2 per cent of
lnjection: 100 µL. dimer (typically 12-24 h). Dissolve its contents in 0.025 M
System suitability: the chromatograms obtained with the phosphate buffer solution pH 7.0 R and dilute with the same
test solution and the reference solution are similar to solution to obtain a concentration of 1.0 mg/mL.
the chromatogram of somatropin digest supplied with Column:
somatropin CRS. - size: l = 0.30 m, 0 = 7.8 mm;
Results: the profile of the chromatogram obtained with - stationary phase: hydrophilic silica gel for chromatography R
the test solution corresponds to that of the chromatogram of a grade suitable for fractionation of globular proteins in
obtained with the reference solution. the relative molecular mass range of 5000 to 150 000.
D. Examine the chromatograms obtained in the assay. Mobile phase: 2-propanol R, 0.063 M phosphate buffer solution
pH 7.0 R (3:97 V/V); filter and <legas.
with the test solution is similar in retention time and size
to the principal peak in the chromatogram obtained with Detection: spectrophotometer at 214 nm.
the reference solution. Injection: 20 µL.
Somatropin for injection EUROPEAN PHARMACOPOEIA 9.5
Relative retention with reference to somatropin monomer Migration: apply a field strength of 217 V/cm (20 kV for
(retention time = 12 min to 17 min): related substances capillaries of 92 cm total length) for 80 min, using CZE buffer
of higher molecular mass = about 0.65; somatropin as the electrolyte in both buffer reservoirs.
dimer = about 0.9. Relative migration with reference to somatropin: deamidated
System suitability: resolution solution: forms = 1.02 to 1.11.
- peak-to-valley ratio: minimum 2.5, where HP = height above System suitability: reference solution:
the baseline of the peak due to the dimer and Hv = height - the electropherogram obtained is similar to the
above the baseline of the lowest point of the curve electropherogram of somatropin supplied with
separating this peak from the peak due to the monomer. somatropin CRS; 2 peaks (Ip 12) eluting prior to the
Limit: principal peak and at least 2 peaks (I 3, 14 ) eluting after the
principal peak are clearly visible.
- sum of the peaks with retention times less than that of the
Note: peak 12 corresponds to the cleaved form and peak ! 4
corresponds to the deamidated forms, eluting as a doublet.
Charged variants. Capillary electrophoresis (2.2.47). Limits:
Test solution (a). Prepare a solution of the substance to be - deamidated forms: maximum 6.5 per cent;
examined containing 1 mg/mL of somatropin. - any other ímpuríty: for each impurity, maximum 2.0 per
Test solution (b ). Mix equal volumes of test solution (a) and cent;
the reference solution. - total: maximum 11.5 per cent.
Reference solution. Dissolve the contents of a vial of Water (2.5.32): maximum 3.0 per cent, unless otherwise
somatropin CRS in water R and dilute with the same solvent justified and authorised.
Bacterial endotoxins (2.6.14): less than 5 IU/mg.
Capillary:
- material: uncoated fused silica; ASSAY
- size: effective length = at least 70 cm, 0 = 50 µm. Size-exclusion chromatography (2.2.30) as described in the
test for dimer and related substances ofhigher molecular mass.
Calculate the content of somatropin (C990 H 1528 N 262 Ü 300S7 ) from
CZE buffer: 13.2 g/L solution of ammonium phosphate R the declared content of C990H 1528N 262 0 300S7 in somatropin CRS.
adjusted to pH 6.0 with phosphoric acid R and filtered.
Detection: spectrophotometer at 200 nm. STORAGE
In a sterile, airtight, tamper-proof container, ata temperature
Set the autosampler to store the samples at 4 ºC during analysis.
of 2 ºC to 8 ºC.
Preconditioning of the capillary: rinse with 1 M sodium
hydroxide for 20 min, with water R for 1O min and with LABELLING
the CZE buffer for 20 min. The label states:
Between-run rinsing: rinse with 0.1 M sodium hydroxide for - the content of somatropin in the container, in milligrams;
2 min and with the CZE buffer for 6 min. - the composition and volume of the liquid to be added for
Note: rinsing times may be adapted according to the length of reconstitution;
the capillary and the equipment used. - the time within which the reconstituted solution shall be
Injection: test solution (a) and the reference solution; under used and the storage conditions during this period;
pressure or vacuum, using the following sequence: sample - the name and quantity of any excipient;
injection for at least 3 s then CZE buffer injection for 1 s. - the storage temperature;
The injection time and pressure may be adapted in order to - that the preparation shall not be shaken during
meet the system suitability criteria. reconstitution.

z
Zolmitriptan ............................................................................ 5759

EUROPEAN PHARMACOPOEIA 9.5 Zolmitriptan
07/2018:2737 Limit:
- impurity A: maximum 0.10 per cent.
Solvent mixture: mobile phase B, mobile phase A (10:90 V!V).
ZOLMITRIPTAN Test solution (a). Dissolve 10.0 mg of the substance to be
examined in the solvent mixture and dilute to 10.0 mL with
the solvent mixture.
Zolmitriptanum Test solution (b ). Dilute 1.0 mL of test solution (a) to 5.0 mL
o
Reference solution (a). Dilute 1.0 mL of test solution (a) to
o~ANH
¡
~~ ~j 1
100.0 mL with the solvent mixture. Dilute 1.0 mL of this
H Reference solution (b ). Dissolve 5 mg of zolmitriptan for system
N-CH3
suitability CRS (containing impurities C, H and I) in the
solvent mixture and dilute to 5.0 mL with the solvent mixture.
H3é
Reference solution (e). Dissolve 10.0 mg of zolmitriptan CRS
C16H21N302 Mr 287.4 in the solvent mixture and dilute to 1O.O mL with the solvent
[139264-17-8] mixture. Dilute 1.0 mL of the solution to 5.0 mL with the
solvent mixture.
DEFINITION
Column:
(45)-4-[ [3-[2-(Dimethylamino )ethyl]-lH-indol-5-yl]methyl]-
1,3-oxazolidin-2-one. - size: l = 0.10 m, 0 = 3.0 mm;
Content: 97.5 per cent to 102.0 per cent (anhydrous substance). - stationary phase: end-capped solid core phenylhexylsilyl
silica gel for chromatography R (2.7 µm);
CHARACTERS - temperature: 20 ºC.
Appearance: white or almost white powder. Mobile phase A: dissolve 2.72 g of potassium dihydrogen
Solubility: slightly soluble or very slightly soluble in water, phosphate R and 0.94 g of sodium hexanesulfonate R in water
freely soluble in methanol, sparingly soluble in acetone. for chromatography R, adjust to pH 2.0 with phosphoric acid R
and dilute to 1000 mL with water for chromatography R;
IDENTIFICATION Mobile phase B: acetonitrile Rl ;
Infrared absorption spectrophotometry (2.2.24). Time Mobile phase A Mobile phase B
Comparison: zolmitriptan CRS. (min) (per cent V!V) (per cent V/V)
TESTS
o - 0.5 90 10
Enantiomeric purity. Liquid chromatography (2.2.29). 0.5 - 4 90 -7 85 10 -7 15
Test solution. Dissolve 25.0 mg of the substance to be 4-8 85 15

examined in the mobile phase and dilute to 25.0 mL with the 8 -9 85 -7 80 15 -7 20
mobile phase.
Reference solution (a). Dilute 1.0 mL of the test solution to 9 - 10 80 20
100.0 mL with the mobile phase. Dilute 1.0 mL of this solution 10 - 12 80 -7 70 20 -7 30
12 - 13 70 30
Reference solution (b ). Dissolve 5 mg of zolmitriptan
impurity A CRS in the mobile phase and dilute to 5.0 mL with Flow rate: 0.8 mL/min.
the mobile phase. To 1.0 mL of the solution add 1.0 mL of
the test solution and dilute to 10.0 mL with the mobile phase.
Dilute 1.0 mL of this solution to SO.O mL with the mobile Injection: 2 µL of test solution (a) and reference solutions (a)
phase. and (b).
Column: Identification of impurities: use the chromatogram supplied
with zolmitriptan for system suitability CRS and the
- size: l = 0.25 m, 0 = 4.6 mm;
chromatogram obtained with reference solution (b) to identify
- stationary phase: amylose derivative of silica gel for chiral the peaks due to impurities C, H and I.
separation R (5 µm);
Relative retention with reference to zolmitriptan
- temperature: 35 ºC. (retention time = about 5 min): impurity H = about 0.97;
Mobile phase: diethylamine R, 2-propanol R, methanol R, impurity I = about 1.1; impurity C = about 2.0.
heptane R (0.1:10:15:75 V/V/V/V). System suitability: reference solution (b):
Flow rate: 1.0 mL/min. - peak-to-valley ratio: minimum 8, where lfp = height above
Detection: spectrophotometer at 285 nm. the baseline of the peak due to impurity H and Hv = height
Injection: 10 µL. above the baseline of the lowest point of the curve
separating this peak from the peak due to zolmitriptan;
Run time: twice the retention time of zolmitriptan.
minimum 1.5, where HP = height above the baseline of the
Relative retention with reference to zolmitriptan (retention peak due to impurity I and Hv = height above the baseline
time= about 7 min): impurity A= 0.7. of the lowest point of the curve separating this peak from
System suitability: reference solution (b): the peak due to zolmitriptan.
- resolution: mínimum 2.0 between the peaks due to Calculation of percentage contents:
impurity A and zolmitriptan. - correction factor: multiply the peak area of impurity C by
Calculation of percentage content: 2.0;
- for impurity A, use the concentration of zolmitriptan in - for each impurity, use the concentration of zolmitriptan in
reference solution (a). reference solution (a).
Zolmitriptan EUROPEAN PHARMACOPOEIA 9.5
Limits: o
o~ANH
~~ ~IÍ : 1
0.10 per cent; H

- total: maximum 0.5 per cent; NH 2
Water (2.5.12): maximum 0.5 per cent, determined on 0.500 g. D. (4S)-4-[[3-(2-aminoethyl)-1H-indol-5-yl]methyl]-1,3-
oxazolidin-2-one,
1.0 g.
ASSAY
lnjection: 2 µL of test solution (b) and reference solution (c).
Calculate the percentage content of C 16 H 21 N 30 2 taking into
account the assigned content of zolmitriptan CRS. E. (4S)-4-[ (4-aminophenyl)methyl]-1,3-oxazolidin-2-one,
IMPURITIES
Specified impurities: A, C.
Control of impurities in substances for pharmaceutical use):
B, D, E, F, G, H, l. F. (2S)-2-amino-3-[3-[2-( dimethylamino )ethyl]-lH-indol-
J-y 11------ -1
c: ..ljjJlVj:'dH-1-Vl,
1
OANH~I
\ _____(
~IÍ
~
H
o
1
N-CH 3
O~)lNH
~ ,,-
~
H3C 1
i IÍ
H
A. (4R)-4-[[3-[2-( dimethylamino)ethyl]-lH-indol-5-
yl]methyl]-1,3-oxazolidin-2-one, HN-CH3
G. (4S)-4-[ [3-[2-(methylamino )ethyl]-lH-indol-5-yl]methyl]-

l,3-oxazolidin-2-one,
B. N,N-dimethyl-2-[5-[[ (4S)-2-oxo-l,3-oxazolidin-4-
yl]methyl]-1H-indol-3-yl]ethan-1-amine N-oxide,
H. (4S)-4-[ (2-methyl-l,2,3,4-tetrahydro-9H-pyrido[3,4-
b]indol-6-yl)methyl]-1 ,3-oxazolidin-2-one,
O~ANH
~ ~¡j 1 C02H
... H i ~
H
N-CH 3
1
H3C
C. (4S,4'S)-4,4' -[[4-(dimethylamino)butane-1,1-
diyl]bis [[3-[2-( dimethylamino )ethyl]- lH-indole-2,5- l. 3-[2-( dimethylamino )ethyl]-5-[[ (4S)-2-oxo-l,3-
diyl]methylene] ]bis( l ,3-oxazolidin-2-one), oxazolidin-4-yl] methyl ]-1 H-indole-2-carboxylic a cid.

INDEX
To aid users the index includes a reference to the supplement in which the latest version of a text can be found.
For example : Alfuzosin hydrochloride...............................................9.1‑4123
means the monograph Alfuzosin hydrochloride can be found on page 4123 of Supplement 9.1.
Note that where no reference to a supplement is made, the text can be found in the principal volume.
English index ..................................................................... 5763 Latin index .............................................................................. 5799

EUROPEAN PHARMACOPOEIA 9.5 Index
Numerics 2.2.7. Optical rotation ..................................................... 9.5-5535

1. General notices............................................................ 9.2-4275 2.2.8. Viscosity ................................................................. 9.4-5099
2.1.1. Droppers ........................................................................... 15 2.2.9. Capillary viscometer method ......................................... 27
2.1.2. Comparative table of porosity of sintered-glass filters.. 15 2.2. Physical and physicochemical methods ........................... 21
2.1.3. Ultraviolet ray lamps for analytical purposes............... 15 2.3.1. Identification reactions of ions and functional
2.1.4. Sieves ................................................................................. 16 groups ...................................................................................... 123
2.1.5. Tubes for comparative tests ............................................ 17 2.3.2. Identification of fatty oils by thin-layer
2.1.6. Gas detector tubes................................................. 9.3-4685 chromatography...................................................................... 126
2.1. Apparatus ............................................................................. 15 2.3.3. Identification of phenothiazines by thin-layer
2.2.10. Viscosity - Rotating viscometer method ..................... 28 chromatography...................................................................... 127
2.2.11. Distillation range ........................................................... 30 2.3.4. Odour .............................................................................. 127
2.2.12. Boiling point................................................................... 31 2.3. Identification...................................................................... 123
2.2.13. Determination of water by distillation ........................ 31 2.4.10. Lead in sugars............................................................... 136
2.2.14. Melting point - capillary method...................... 9.1-4037 2.4.11. Phosphates .................................................................... 136
2.2.15. Melting point - open capillary method ....................... 32 2.4.12. Potassium ...................................................................... 136
2.2.16. Melting point - instantaneous method........................ 33 2.4.13. Sulfates........................................................................... 136
2.2.17. Drop point ...................................................................... 33 2.4.14. Sulfated ash ................................................................... 136
2.2.18. Freezing point................................................................. 34 2.4.15. Nickel in polyols........................................................... 137
2.2.19. Amperometric titration................................................. 35 2.4.16. Total ash ........................................................................ 137
2.2.1. Clarity and degree of opalescence of liquids ..... 9.2-4285 2.4.17. Aluminium ................................................................... 137
2.2.20. Potentiometric titration ................................................ 35 2.4.18. Free formaldehyde ....................................................... 137
2.2.21. Fluorimetry..................................................................... 36 2.4.19. Alkaline impurities in fatty oils.................................. 138
2.2.22. Atomic emission spectrometry .................................... 36 2.4.1. Ammonium .................................................................... 131
2.2.23. Atomic absorption spectrometry ................................. 37 2.4.20. Determination of elemental impurities............ 9.5-5539
2.2.24. Absorption spectrophotometry, infrared.................... 39 2.4.21. Foreign oils in fatty oils by thin-layer
2.2.25. Absorption spectrophotometry, ultraviolet and chromatography...................................................................... 141
visible ......................................................................................... 41 2.4.22. Composition of fatty acids by gas chromatography.. 141
2.2.26. Paper chromatography .................................................. 42 2.4.23. Sterols in fatty oils........................................................ 144
2.2.27. Thin-layer chromatography.......................................... 43 2.4.24. Identification and control of residual solvents ......... 146
2.2.28. Gas chromatography...................................................... 44 2.4.25. Ethylene oxide and dioxan.......................................... 151
2.2.29. Liquid chromatography................................................. 46 2.4.26. N,N-Dimethylaniline ................................................... 152
2.2.2. Degree of coloration of liquids....................................... 22 2.4.27. Heavy metals in herbal drugs and herbal drug
2.2.30. Size-exclusion chromatography ................................... 47 preparations ............................................................................ 152
2.2.31. Electrophoresis ............................................................... 48 2.4.28. 2-Ethylhexanoic acid ................................................... 154
2.2.32. Loss on drying..................................................... 9.4-5099 2.4.29. Composition of fatty acids in oils rich in omega-3
2.2.33. Nuclear magnetic resonance spectrometry ................ 54 acids.......................................................................................... 155
2.2.34. Thermal analysis ............................................................ 57 2.4.2. Arsenic.................................................................... 9.4-5103
2.2.35. Osmolality....................................................................... 59 2.4.30. Ethylene glycol and diethylene glycol in ethoxylated
2.2.36. Potentiometric determination of ionic substances................................................................................ 157
concentration using ion-selective electrodes ........................ 60 2.4.31. Nickel in hydrogenated vegetable oils .............. 9.4-5103
2.2.37. X-ray fluorescence spectrometry ...................... 9.3-4689 2.4.32. Total cholesterol in oils rich in omega-3 acids ......... 157
2.2.38. Conductivity ................................................................... 62 2.4.3. Calcium ........................................................................... 132
2.2.39. Molecular mass distribution in dextrans .................... 62 2.4.4. Chlorides......................................................................... 132
2.2.3. Potentiometric determination of pH ............................. 24 2.4.5. Fluorides.......................................................................... 132
2.2.40. Near-infrared spectroscopy .......................................... 64 2.4.6. Magnesium ..................................................................... 132
2.2.41. Circular dichroism......................................................... 69 2.4.7. Magnesium and alkaline-earth metals ........................ 133
2.2.42. Density of solids............................................................. 70 2.4.8. Heavy metals................................................................... 133
2.2.43. Mass spectrometry......................................................... 71 2.4.9. Iron .................................................................................. 136
2.2.44. Total organic carbon in water for pharmaceutical 2,4-Dichlorobenzyl alcohol ................................................... 2242
use............................................................................................... 73 2.4. Limit tests........................................................................... 131
2.2.45. Supercritical fluid chromatography ............................. 74 2.5.10. Oxygen-flask method .................................................. 164
2.2.46. Chromatographic separation techniques ......... 9.2-4286 2.5.11. Complexometric titrations.......................................... 164
2.2.47. Capillary electrophoresis............................................... 81 2.5.12. Water : semi-micro determination .................... 9.4-5107
2.2.48. Raman spectroscopy...................................................... 86 2.5.13. Aluminium in adsorbed vaccines .............................. 165
2.2.49. Falling ball and automatic rolling ball viscometer 2.5.14. Calcium in adsorbed vaccines .................................... 166
methods .......................................................................... 9.3-4690 2.5.15. Phenol in immunosera and vaccines......................... 166
2.2.4. Approximate pH of solutions ......................................... 25 2.5.16. Protein in polysaccharide vaccines ............................ 166
2.2.54. Isoelectric focusing ........................................................ 89 2.5.17. Nucleic acids in polysaccharide vaccines .................. 166
2.2.55. Peptide mapping ............................................................ 90 2.5.18. Phosphorus in polysaccharide vaccines .................... 166
2.2.56. Amino acid analysis............................................ 9.3-4691 2.5.19. O-Acetyl in polysaccharide vaccines ......................... 167
2.2.57. Inductively coupled plasma-atomic emission 2.5.1. Acid value........................................................................ 161
spectrometry ........................................................................... 100 2.5.20. Hexosamines in polysaccharide vaccines.................. 167
2.2.58. Inductively coupled plasma-mass spectrometry...... 101 2.5.21. Methylpentoses in polysaccharide vaccines.............. 167
2.2.59. Glycan analysis of glycoproteins ................................ 103 2.5.22. Uronic acids in polysaccharide vaccines ................... 168
2.2.5. Relative density................................................................. 25 2.5.23. Sialic acid in polysaccharide vaccines ....................... 168
2.2.61. Characterisation of crystalline solids by 2.5.24. Carbon dioxide in gases .............................................. 168
microcalorimetry and solution calorimetry........................ 109 2.5.25. Carbon monoxide in gases ......................................... 169
2.2.64. Peptide identification by nuclear magnetic resonance 2.5.26. Nitrogen monoxide and nitrogen dioxide in gases.. 170
spectrometry ........................................................................... 112 2.5.27. Oxygen in gases............................................................ 170
2.2.65. Voltametric titration .................................................... 112 2.5.28. Water in gases............................................................... 170
2.2.66. Detection and measurement of radioactivity ........... 113 2.5.29. Sulfur dioxide ............................................................... 171
2.2.6. Refractive index................................................................ 26 2.5.2. Ester value....................................................................... 161
2.5.30. Oxidising substances ................................................... 171
Index EUROPEAN PHARMACOPOEIA 9.5
2.5.31. Ribose in polysaccharide vaccines ............................. 171 2.7.1. Immunochemical methods........................................... 239
2.5.32. Water : micro determination.............................. 9.4-5107 2.7.20. In vivo assay of poliomyelitis vaccine (inactivated).. 267
2.5.33. Total protein ................................................................. 172 2.7.21. Assay of human von Willebrand factor..................... 268
2.5.34. Acetic acid in synthetic peptides................................ 175 2.7.22. Assay of human coagulation factor XI ...................... 269
2.5.35. Nitrous oxide in gases ................................................. 176 2.7.23. Numeration of CD34/CD45+ cells in
2.5.36. Anisidine value............................................................. 176 haematopoietic products ....................................................... 269
2.5.37. Methyl, ethyl and isopropyl methanesulfonate in 2.7.24. Flow cytometry ............................................................ 271
methanesulfonic acid ............................................................. 176 2.7.25. Assay of human plasmin inhibitor............................. 272
2.5.38. Methyl, ethyl and isopropyl methanesulfonate in active 2.7.27. Flocculation value (Lf) of diphtheria and tetanus toxins
substances................................................................................ 177 and toxoids (Ramon assay) ................................................... 273
2.5.39. Methanesulfonyl chloride in methanesulfonic 2.7.28. Colony-forming cell assay for human
acid ........................................................................................... 178 haematopoietic progenitor cells............................................ 274
2.5.3. Hydroxyl value ............................................................... 161 2.7.29. Nucleated cell count and viability.............................. 275
2.5.40. Methyl, ethyl and isopropyl toluenesulfonate in active 2.7.2. Microbiological assay of antibiotics.................... 9.3-4719
substances................................................................................ 179 2.7.30. Assay of human protein C .......................................... 276
2.5.41. Methyl, ethyl and isopropyl benzenesulfonate in active 2.7.31. Assay of human protein S ........................................... 277
substances................................................................................ 180 2.7.32. Assay of human α-1-proteinase inhibitor ................. 278
2.5.4. Iodine value .................................................................... 161 2.7.34. Assay of human C1-esterase inhibitor ...................... 278
2.5.5. Peroxide value................................................................. 162 2.7.35. Immunonephelometry for vaccine component
2.5.6. Saponification value....................................................... 163 assay ......................................................................................... 279
2.5.7. Unsaponifiable matter ................................................... 163 2.7.4. Assay of human coagulation factor VIII ..................... 246
2.5.8. Determination of primary aromatic amino- 2.7.5. Assay of heparin.................................................... 9.1-4049
nitrogen.................................................................................... 163 2.7.6. Assay of diphtheria vaccine (adsorbed) ...................... 247
2.5.9. Determination of nitrogen by sulfuric acid 2.7.7. Assay of pertussis vaccine (whole cell)........................ 252
digestion .................................................................................. 164 2.7.8. Assay of tetanus vaccine (adsorbed)............................ 253
2.5. Assays ................................................................................. 161 2.7.9. Test for Fc function of immunoglobulin............ 9.3-4724
2.6.10. Histamine...................................................................... 194 2.7. Biological assays ................................................................ 239
2.6.11. Depressor substances................................................... 195 2.8.10. Solubility in alcohol of essential oils.......................... 284
2.6.12. Microbiological examination of non-sterile products : 2.8.11. Assay of 1,8-cineole in essential oils.......................... 285
microbial enumeration tests.................................................. 195 2.8.12. Essential oils in herbal drugs...................................... 285
2.6.13. Microbiological examination of non-sterile products : 2.8.13. Pesticide residues ......................................................... 286
test for specified micro-organisms ....................................... 199 2.8.14. Tannins in herbal drugs .............................................. 288
2.6.14. Bacterial endotoxins ........................................... 9.3-4707 2.8.15. Bitterness value............................................................. 288
2.6.15. Prekallikrein activator ................................................. 208 2.8.16. Dry residue of extracts ................................................ 289
2.6.16. Tests for extraneous agents in viral vaccines for human 2.8.17. Loss on drying of extracts........................................... 289
use.................................................................................... 9.4-5111 2.8.18. Determination of aflatoxin B1 in herbal drugs ......... 289
2.6.17. Test for anticomplementary activity of 2.8.1. Ash insoluble in hydrochloric acid .............................. 283
immunoglobulin............................................................ 9.3-4713 2.8.20. Herbal drugs : sampling and sample preparation .... 291
2.6.18. Test for neurovirulence of live virus vaccines .......... 212 2.8.21. Test for aristolochic acids in herbal drugs ................ 292
2.6.1. Sterility ............................................................................ 185 2.8.22. Determination of ochratoxin A in herbal drugs ...... 294
2.6.20. Anti-A and anti-B haemagglutinins .......................... 213 2.8.23. Microscopic examination of herbal drugs ................ 295
2.6.21. Nucleic acid amplification techniques....................... 214 2.8.25. High-performance thin-layer chromatography of
2.6.22. Activated coagulation factors ..................................... 219 herbal drugs and herbal drug preparations ........................ 295
2.6.24. Avian viral vaccines : tests for extraneous agents in 2.8.2. Foreign matter ................................................................ 283
seed lots ................................................................................... 219 2.8.3. Stomata and stomatal index.......................................... 283
2.6.25. Avian live virus vaccines : tests for extraneous agents in 2.8.4. Swelling index................................................................. 284
batches of finished product ................................................... 222 2.8.5. Water in essential oils .................................................... 284
2.6.26. Test for anti-D antibodies in human immunoglobu- 2.8.6. Foreign esters in essential oils ...................................... 284
lin.............................................................................................. 225 2.8.7. Fatty oils and resinified essential oils in essential
2.6.27. Microbiological examination of cell-based oils ............................................................................................ 284
preparations ................................................................... 9.2-4297 2.8.8. Odour and taste of essential oils .................................. 284
2.6.2. Mycobacteria .................................................................. 188 2.8.9. Residue on evaporation of essential oils ..................... 284
2.6.30. Monocyte-activation test.................................... 9.2-4299 2.8. Methods in pharmacognosy ............................................ 283
2.6.31. Microbiological examination of herbal medicinal 2.9.10. Ethanol content ............................................................ 315
products for oral use and extracts used in their 2.9.11. Test for methanol and 2-propanol ............................. 318
preparation .............................................................................. 232 2.9.12. Sieve test........................................................................ 319
2.6.33. Residual pertussis toxin and irreversibility of pertussis 2.9.14. Specific surface area by air permeability.......... 9.1-4053
toxoid ....................................................................................... 235 2.9.16. Flowability..................................................................... 321
2.6.34. Host-cell protein assays...................................... 9.1-4041 2.9.17. Test for extractable volume of parenteral
2.6.7. Mycoplasmas .................................................................. 188 preparations ............................................................................ 322
2.6.8. Pyrogens.......................................................................... 193 2.9.18. Preparations for inhalation : aerodynamic assessment
2.6.9. Abnormal toxicity .......................................................... 194 of fine particles........................................................................ 323
2.6. Biological tests ................................................................... 185 2.9.19. Particulate contamination : sub-visible particles...... 335
2.7.10. Assay of human coagulation factor VII .................... 258 2.9.1. Disintegration of tablets and capsules......................... 299
2.7.11. Assay of human coagulation factor IX ...................... 259 2.9.20. Particulate contamination : visible particles ............. 337
2.7.12. Assay of heparin in coagulation factors .................... 259 2.9.22. Softening time determination of lipophilic
2.7.13. Assay of human anti-D immunoglobulin ................. 260 suppositories ........................................................................... 338
2.7.14. Assay of hepatitis A vaccine ....................................... 262 2.9.23. Gas pycnometric density of solids ............................. 339
2.7.15. Assay of hepatitis B vaccine (rDNA) ......................... 263 2.9.25. Dissolution test for medicated chewing gums ......... 340
2.7.16. Assay of pertussis vaccine (acellular) ........................ 263 2.9.26. Specific surface area by gas adsorption ..................... 344
2.7.17. Assay of human antithrombin III .............................. 265 2.9.27. Uniformity of mass of delivered doses from multidose
2.7.18. Assay of human coagulation factor II ....................... 266 containers ................................................................................ 347
2.7.19. Assay of human coagulation factor X........................ 266 2.9.29. Intrinsic dissolution..................................................... 347

2.9.2. Disintegration of suppositories and pessaries ............ 301 3.2.6. Sets for the transfusion of blood and blood
2.9.31. Particle size analysis by laser light diffraction.......... 349 components ............................................................................. 433
2.9.32. Porosity and pore-size distribution of solids by 3.2.8. Sterile single-use plastic syringes ................................. 434
mercury porosimetry ............................................................. 352 3.2.9. Rubber closures for containers for aqueous
2.9.33. Characterisation of crystalline and partially crystalline parenteral preparations, for powders and for
solids by X-ray powder diffraction (XRPD) ....................... 354 freeze-dried powders..................................................... 9.5-5545
2.9.34. Bulk density and tapped density of powders............ 359 3.2. Containers.......................................................................... 423
2.9.35. Powder fineness............................................................ 362 3-O-Desacyl-4-monophosphoryl lipid A............................ 2207
2.9.36. Powder flow .................................................................. 362 4.1.1. Reagents ................................................................. 9.4-5131
2.9.37. Optical microscopy...................................................... 365 4.1.1. Reagents ................................................................. 9.5-5549
2.9.38. Particle-size distribution estimation by analytical 4.1.2. Standard solutions for limit tests ........................ 9.4-5247
sieving ...................................................................................... 367 4.1.3. Buffer solutions ..................................................... 9.4-5252
2.9.39. Water-solid interactions : determination of 4.1.3. Buffer solutions ..................................................... 9.5-5549
sorption-desorption isotherms and of water activity......... 369 4.1. Reagents, standard solutions, buffer solutions ..... 9.5-5549
2.9.3. Dissolution test for solid dosage forms ....................... 302 4.2.1. Primary standards for volumetric solutions...... 9.4-5258
2.9.40. Uniformity of dosage units ................................ 9.1-4055 4.2.2. Volumetric solutions............................................. 9.4-5258
2.9.41. Friability of granules and spheroids .......................... 375 4.2.2. Volumetric solutions............................................. 9.5-5550
2.9.42. Dissolution test for lipophilic solid dosage forms ... 377 4.2. Volumetric analysis.................................................. 9.4-5258
2.9.43. Apparent dissolution ................................................... 377 4-Aminobenzoic acid ............................................................. 1702
2.9.44. Preparations for nebulisation : characterisation....... 378 4. Reagents........................................................................ 9.4-5131
2.9.45. Wettability of porous solids including powders....... 381 5.10. Control of impurities in substances for pharmaceutical
2.9.47. Demonstration of uniformity of dosage units using use............................................................................................. 723
large sample sizes.................................................................... 384 5.1.10. Guidelines for using the test for bacterial
2.9.4. Dissolution test for transdermal patches .................... 309 endotoxins ............................................................................... 593
2.9.5. Uniformity of mass of single-dose preparations ........ 311 5.1.11. Determination of bactericidal, fungicidal or yeasticidal
2.9.6. Uniformity of content of single-dose preparations.... 312 activity of antiseptic medicinal products.................... 9.2-4348
2.9.7. Friability of uncoated tablets ........................................ 312 5.11. Characters section in monographs ............................... 729
2.9.8. Resistance to crushing of tablets .................................. 313 5.1.1. Methods of preparation of sterile products ....... 9.2-4333
2.9.9. Measurement of consistency by penetrometry .......... 313 5.1.2. Biological indicators and related microbial preparations
2.9. Pharmaceutical technical procedures ............................. 299 used in the manufacture of sterile products............... 9.2-4336
3.1.10. Materials based on non-plasticised poly(vinyl chloride) 5.12. Reference standards ............................................... 9.5-5563
for containers for non-injectable, aqueous solutions......... 410 5.1.3. Efficacy of antimicrobial preservation ........................ 577
3.1.11. Materials based on non-plasticised poly(vinyl 5.14. Gene transfer medicinal products for human use ...... 739
chloride) for containers for solid dosage forms for oral 5.1.4. Microbiological quality of non-sterile pharmaceutical
administration......................................................................... 412 preparations and substances for pharmaceutical use......... 579
3.1.1.1. Materials based on plasticised poly(vinyl chloride) for 5.1.5. Application of the F0 concept to steam sterilisation of
containers for human blood and blood components......... 391 aqueous preparations ............................................................. 580
3.1.1.2. Materials based on plasticised poly(vinyl chloride) for 5.15. Functionality-related characteristics of
tubing used in sets for the transfusion of blood and blood excipients ........................................................................ 9.2-4411
components ............................................................................. 393 5.1.6. Alternative methods for control of microbiological
3.1.13. Plastic additives ............................................................ 414 quality ............................................................................. 9.2-4339
3.1.14. Materials based on plasticised poly(vinyl chloride) 5.16. Crystallinity ..................................................................... 757
for containers for aqueous solutions for intravenous 5.17.1. Recommendations on dissolution testing................. 761
infusion .................................................................................... 416 5.17. Recommendations on methods for dosage forms
3.1.15. Polyethylene terephthalate for containers for testing....................................................................................... 761
preparations not for parenteral use ...................................... 419 5.1.7. Viral safety ...................................................................... 591
3.1.1. Materials for containers for human blood and blood 5.1.8. Microbiological quality of herbal medicinal products for
components ............................................................................. 391 oral use and extracts used in their preparation .................. 591
3.1.3. Polyolefins .............................................................. 9.4-5117 5.19. Extemporaneous preparation of radiopharmaceuti-
3.1.4. Polyethylene without additives for containers cals ............................................................................................ 767
for parenteral preparations and for ophthalmic 5.1.9. Guidelines for using the test for sterility..................... 592
preparations ................................................................... 9.2-4310 5.1. General texts on microbiology ........................................ 575
3.1.5. Polyethylene with additives for containers for parenteral 5.20. Elemental impurities ............................................. 9.3-4759
preparations and for ophthalmic preparations .......... 9.4-5120 5.2.11. Carrier proteins for the production of conjugated
3.1.6. Polypropylene for containers and closures for parenteral polysaccharide vaccines for human use............................... 626
preparations and ophthalmic preparations ................ 9.4-5123 5.2.12. Raw materials of biological origin for the production
3.1.7. Poly(ethylene - vinyl acetate) for containers and tubing of cell-based and gene therapy medicinal products ........... 627
for total parenteral nutrition preparations ................. 9.2-4318 5.2.13. Healthy chicken flocks for the production of inactivated
3.1.8. Silicone oil used as a lubricant ..................................... 408 vaccines for veterinary use .................................................... 630
3.1.9. Silicone elastomer for closures and tubing ................. 409 5.2.14. Substitution of in vivo method(s) by in vitro method(s)
3.1. Materials used for the manufacture of containers ........ 391 for the quality control of vaccines ............................... 9.3-4737
3.2.1. Glass containers for pharmaceutical use..................... 423 5.21. Chemometric methods applied to analytical data ...... 783
3.2.2.1. Plastic containers for aqueous solutions for 5.2.1. Terminology used in monographs on biological
infusion .................................................................................... 429 products ................................................................................... 599
3.2.2. Plastic containers and closures for pharmaceutical 5.2.2. Chicken flocks free from specified pathogens for the
use............................................................................................. 428 production and quality control of vaccines......................... 599
3.2.3. Sterile plastic containers for human blood and 5.22. Names of herbal drugs used in traditional Chinese
blood components .................................................................. 430 medicine ......................................................................... 9.4-5283
3.2.4. Empty sterile containers of plasticised poly(vinyl 5.2.3. Cell substrates for the production of vaccines for human
chloride) for human blood and blood components........... 432 use.................................................................................... 9.3-4733
3.2.5. Sterile containers of plasticised poly(vinyl chloride) for 5.23. Monographs on herbal drug extracts (information
human blood containing anticoagulant solution ............... 432 chapter) .................................................................................... 807
5.2.4. Cell cultures for the production of veterinary Aerodynamic assessment of fine particles in preparations for
vaccines.................................................................................... 606 inhalation (2.9.18.) ................................................................. 323
5.24. Chemical imaging .................................................. 9.3-4767 Aflatoxin B1 in herbal drugs, determination of (2.8.18.) ..... 289
5.2.5. Substances of animal origin for the production of Agar .......................................................................................... 1230
immunological veterinary medicinal products .................. 608 Agaricus phalloides for homoeopathic preparations .. 9.5-5621
5.2.6. Evaluation of safety of veterinary vaccines and Agnus castus fruit ................................................................... 1231
immunosera ........................................................................... 610 Agnus castus fruit dry extract ............................................... 1232
5.2.7. Evaluation of efficacy of veterinary vaccines and Agrimony................................................................................. 1233
immunosera ............................................................................ 612 Air, medicinal ......................................................................... 1653
5.2.8. Minimising the risk of transmitting animal spongiform Air, synthetic medicinal......................................................... 1655
encephalopathy agents via human and veterinary medicinal Akebia stem ............................................................................. 1234
products ................................................................................... 613 Alanine..................................................................................... 1656
5.2.9. Evaluation of safety of each batch of immunosera for Albendazole...................................................................... 9.4-5321
veterinary use.......................................................................... 625 Albumin solution, human ..................................................... 2660
5.2. General texts on biological products .............................. 599 Alchemilla................................................................................ 1235
5.3. Statistical analysis of results of biological assays and Alcoholimetric tables (5.5.) ..................................................... 675
tests.................................................................................. 9.2-4353 Alcuronium chloride.............................................................. 1658
5.4. Residual solvents ...................................................... 9.5-5553 Alendronate trihydrate, sodium ........................................... 3559
5.5. Alcoholimetric tables........................................................ 675 Alexandrian senna pods ........................................................ 1518
5.6. Assay of interferons .......................................................... 689 Alfacalcidol.............................................................................. 1660
5.7. Table of physical characteristics of radionuclides Alfadex .............................................................................. 9.3-4838
mentioned in the European Pharmacopoeia ............. 9.2-4385 Alfentanil hydrochloride ....................................................... 1662
5.8. Pharmacopoeial harmonisation ............................. 9.4-5265 Alfuzosin hydrochloride ................................................. 9.1-4123
5.9. Polymorphism ................................................................... 719 Alginate, sodium..................................................................... 3560
Alginic acid.............................................................................. 1665
A Alimemazine hemitartrate..................................................... 1666
Abacavir sulfate....................................................................... 1619 Alkaline-earth metals and magnesium (2.4.7.)..................... 133
Abbreviations and symbols (1.) ..................................... 9.2-4275 Alkaline impurities in fatty oils (2.4.19.) ............................... 138
Abnormal toxicity (2.6.9.)........................................................ 194 Allantoin .................................................................................. 1667
Absorption spectrophotometry, infrared (2.2.24.)................. 39 Allergen products ..................................................................... 811
Absorption spectrophotometry, ultraviolet and visible Allergen products, animal epithelia and outgrowths for... 1736
(2.2.25.) ...................................................................................... 41 Allergen products, Hymenoptera venoms for..................... 2732
Acacia ....................................................................................... 1229 Allergen products, mites for.................................................. 3077
Acacia, spray-dried ................................................................. 1620 Allergen products, moulds for .............................................. 3094
Acamprosate calcium ............................................................. 1621 Allergen products, pollens for............................................... 3362
Acanthopanax bark ......................................................... 9.2-4447 Allium sativum for homoeopathic preparations ................ 1590
Acarbose .................................................................................. 1622 Allopurinol .............................................................................. 1668
Acebutolol hydrochloride ...................................................... 1624 all-rac-α-Tocopherol............................................................... 3800
Aceclofenac....................................................................... 9.3-4835 all-rac-α-Tocopheryl acetate.................................................. 3802
Acemetacin .............................................................................. 1628 Almagate .................................................................................. 1670
Acesulfame potassium............................................................ 1629 Almond oil, refined ................................................................ 1671
Acetate trihydrate, sodium .................................................... 3558 Almond oil, virgin .................................................................. 1671
Acetazolamide......................................................................... 1630 Aloes, Barbados ...................................................................... 1236
Acetic acid, glacial .................................................................. 1632 Aloes, Cape.............................................................................. 1237
Acetic acid in synthetic peptides (2.5.34.) ............................. 175 Aloes dry extract, standardised............................................. 1238
Acetone .................................................................................... 1632 Alovudine (18F) injection ....................................................... 1127
Acetylcholine chloride ........................................................... 1633 Alphacyclodextrin ........................................................... 9.3-4838
Acetylcysteine.......................................................................... 1634 Alprazolam .............................................................................. 1672
β-Acetyldigoxin....................................................................... 1635 Alprenolol hydrochloride ...................................................... 1674
Acetylene intermix (1 per cent) in nitrogen................. 9.3-4836 Alprostadil ............................................................................... 1675
Acetylsalicylic acid ................................................................. 1638 Alteplase for injection ............................................................ 1677
Acetyltryptophan, N- ............................................................. 1639 Alternative methods for control of microbiological quality
Acetyltyrosine, N-................................................................... 1641 (5.1.6.) ............................................................................. 9.2-4339
Aciclovir................................................................................... 1642 Altizide..................................................................................... 1681
Acidum picrinicum for homoeopathic preparations ......... 1587 Alum......................................................................................... 1682
Acidum succinicum for homoeopathic preparations.. 9.5-5621 Aluminium (2.4.17.)................................................................. 137
Acid value (2.5.1.)..................................................................... 161 Aluminium chloride hexahydrate......................................... 1682
Acitretin ............................................................................ 9.5-5627 Aluminium hydroxide, hydrated, for adsorption ............... 1682
Actinobacillosis vaccine (inactivated), porcine .................. 1085 Aluminium in adsorbed vaccines (2.5.13.)............................ 165
Activated charcoal .................................................................. 2025 Aluminium magnesium silicate ..................................... 9.3-4839
Activated coagulation factors (2.6.22.)................................... 219 Aluminium oxide, hydrated .................................................. 1684
Adapalene ................................................................................ 1646 Aluminium phosphate gel ..................................................... 1685
Additives, plastic (3.1.13.) ....................................................... 414 Aluminium phosphate, hydrated.......................................... 1686
Adenine.................................................................................... 1647 Aluminium sodium silicate ................................................... 1686
Adeno-associated-virus vectors for human use .................... 748 Aluminium stearate................................................................ 1687
Adenosine ................................................................................ 1648 Aluminium sulfate.................................................................. 1689
Adenovirus vaccine (inactivated), canine............................ 1027 Alverine citrate................................................................. 9.3-4841
Adenovirus vaccine (live), canine......................................... 1028 Amantadine hydrochloride ................................................... 1691
Adipic acid............................................................................... 1649 Ambroxol hydrochloride ................................................ 9.2-4499
Adrenaline ............................................................................... 1650 Amfetamine sulfate ................................................................ 1694
Adrenaline tartrate ................................................................. 1652 Amidotrizoate, sodium .......................................................... 3561
Adsorption, gas, specific surface area by (2.9.26.)................ 344 Amidotrizoic acid dihydrate ................................................. 1694
Amikacin ................................................................................. 1696

Amikacin sulfate ..................................................................... 1698 Anticoagulant and preservative solutions for human blood
Amiloride hydrochloride dihydrate .............................. 9.2-4500 ................................................................................................ 1738
Amino acid analysis (2.2.56.) ......................................... 9.3-4691 Anticomplementary activity of immunoglobulin
Aminobenzoic acid, 4- ........................................................... 1702 (2.6.17.) ........................................................................... 9.3-4713
Aminocaproic acid ................................................................. 1703 Anti-D antibodies in human immunoglobulin, test for
Aminoglutethimide ................................................................ 1704 (2.6.26.) .................................................................................... 225
Aminophylline ........................................................................ 3752 Anti-D immunoglobulin for intravenous administration,
Aminophylline hydrate .......................................................... 3754 human .................................................................................... 2663
Aminosalicylate dihydrate, sodium...................................... 3562 Anti-D immunoglobulin, human ......................................... 2662
Amiodarone hydrochloride................................................... 1706 Anti-D immunoglobulin, human, assay of (2.7.13.) ............ 260
Amisulpride............................................................................. 1707 Antimicrobial preservation, efficacy of (5.1.3.) .................... 577
Amitriptyline hydrochloride ................................................. 1709 Antiseptic medicinal products, determination of bactericidal,
Amlodipine besilate................................................................ 1710 fungicidal or yeasticidal activity of (5.1.11.) .............. 9.2-4348
Ammonia (13N) injection....................................................... 1129 Antiserum, European viper venom ...................................... 1119
Ammonia solution, concentrated ......................................... 1712 Antithrombin III concentrate, human ................................. 2664
Ammonio methacrylate copolymer (type A)...................... 1712 Antithrombin III, human, assay of (2.7.17.) ......................... 265
Ammonio methacrylate copolymer (type B) ...................... 1713 Anti-T lymphocyte immunoglobulin for human use,
Ammonium (2.4.1.).................................................................. 131 animal .................................................................................... 1741
Ammonium bromide ............................................................. 1714 Apis for homoeopathic preparations.................................... 1592
Ammonium carbonicum for homoeopathic preparations.. 9.3- Apomorphine hydrochloride hemihydrate ......................... 1744
4827 Apparatus (2.1.) .......................................................................... 15
Ammonium chloride.............................................................. 1715 Apparent dissolution (2.9.43.)................................................. 377
Ammonium glycyrrhizate ..................................................... 1716 Application of the F0 concept to steam sterilisation of aqueous
Ammonium hydrogen carbonate ......................................... 1717 preparations (5.1.5.) ............................................................... 580
Amobarbital ............................................................................ 1717 Approximate pH of solutions (2.2.4.)....................................... 25
Amobarbital sodium .............................................................. 1718 Aprepitant................................................................................ 1746
Amomum fruit........................................................................ 1239 Aprotinin ................................................................................. 1747
Amomum fruit, round ........................................................... 1502 Aprotinin concentrated solution .......................................... 1749
Amorolfine hydrochloride..................................................... 1719 Arachis oil, hydrogenated...................................................... 1752
Amoxicillin sodium................................................................ 1720 Arachis oil, refined ................................................................. 1752
Amoxicillin trihydrate............................................................ 1723 Arginine ................................................................................... 1753
Amperometric titration (2.2.19.) .............................................. 35 Arginine aspartate .................................................................. 1754
Amphotericin B ...................................................................... 1725 Arginine hydrochloride ......................................................... 1755
Ampicillin................................................................................ 1727 Argon ....................................................................................... 1756
Ampicillin sodium.................................................................. 1729 Aripiprazole............................................................................. 1757
Ampicillin trihydrate.............................................................. 1731 Aristolochic acids in herbal drugs, test for (2.8.21) ............. 292
Amylmetacresol ...................................................................... 1733 Arnica flower........................................................................... 1251
Anacardium for homoeopathic preparations...................... 1591 Arnica tincture........................................................................ 1253
Anaemia vaccine (live), chicken, infectious ................. 9.5-5598 Arsenic (2.4.2.)................................................................. 9.4-5103
Anaesthetic ether .................................................................... 2422 Arsenicum album for homoeopathic preparations ..... 9.5-5623
Analysis, thermal (2.2.34.)......................................................... 57 Arsenious trioxide for homoeopathic preparations .... 9.5-5623
Analytical sieving, particle-size distribution estimation by Articaine hydrochloride......................................................... 1758
(2.9.38.) .................................................................................... 367 Artichoke leaf ................................................................... 9.4-5301
Anamirta cocculus for homoeopathic preparations .......... 1597 Artichoke leaf dry extract............................................... 9.4-5302
Anastrozole....................................................................... 9.3-4842 Ascorbate, calcium ................................................................. 1911
Andrographis herb .......................................................... 9.3-4807 Ascorbate, sodium .................................................................. 3563
Anemarrhena asphodeloides rhizome ................................. 1241 Ascorbic acid .................................................................... 9.3-4843
Angelica archangelica root .................................................... 1242 Ascorbyl palmitate.................................................................. 1762
Angelica dahurica root.................................................... 9.3-4808 Ash insoluble in hydrochloric acid (2.8.1.) ........................... 283
Angelica pubescens root ................................................. 9.3-4810 Ash leaf .................................................................................... 1257
Angelica sinensis root ..................................................... 9.1-4100 Ash, sulfated (2.4.14.)............................................................... 136
Animal anti-T lymphocyte immunoglobulin for human Ash, total (2.4.16.) .................................................................... 137
use........................................................................................... 1741 Asparagine monohydrate....................................................... 1762
Animal epithelia and outgrowths for allergen products.... 1736 Aspartame................................................................................ 1763
Animal immunosera for human use ...................................... 821 Aspartic acid..................................................................... 9.3-4845
Animal spongiform encephalopathies, products with risk of Assay of 1,8-cineole in essential oils (2.8.11.) ....................... 285
transmitting agents of ............................................................ 832 Assay of diphtheria vaccine (adsorbed) (2.7.6.).................... 247
Animal spongiform encephalopathy agents, minimising the Assay of heparin (2.7.5.) ................................................. 9.1-4049
risk of transmitting via human and veterinary medicinal Assay of heparin in coagulation factors (2.7.12.) ................. 259
products (5.2.8.)...................................................................... 613 Assay of hepatitis A vaccine (2.7.14.)..................................... 262
Aniseed ............................................................................. 9.2-4448 Assay of hepatitis B vaccine (rDNA) (2.7.15.) ...................... 263
Anise oil ................................................................................... 1248 Assay of human α-1-proteinase inhibitor (2.7.32.) .............. 278
Anisidine value (2.5.36.) .......................................................... 176 Assay of human anti-D immunoglobulin (2.7.13.) .............. 260
Antazoline hydrochloride...................................................... 1737 Assay of human antithrombin III (2.7.17.)............................ 265
Anthrax spore vaccine (live) for veterinary use.................. 1003 Assay of human C1-esterase inhibitor (2.7.34.).................... 278
Anthrax vaccine for human use (adsorbed, prepared from Assay of human coagulation factor II (2.7.18.)..................... 266
culture filtrates)....................................................................... 893 Assay of human coagulation factor IX (2.7.11.) ................... 259
Anti-A and anti-B haemagglutinins (2.6.20.)........................ 213 Assay of human coagulation factor VII (2.7.10.).................. 258
Antibiotics, microbiological assay of (2.7.2.) ............... 9.3-4719 Assay of human coagulation factor VIII (2.7.4.) .................. 246
Antibodies (anti-D) in human immunoglobulin, test for Assay of human coagulation factor X (2.7.19.)..................... 266
(2.6.26.) .................................................................................... 225 Assay of human coagulation factor XI (2.7.22.) ................... 269
Antibodies for human use, monoclonal ................................ 826 Assay of human plasmin inhibitor (2.7.25.).......................... 272
Assay of human protein C (2.7.30.)........................................ 276
Assay of human protein S (2.7.31.) ........................................ 277 Barium chloratum for homoeopathic preparations ........... 1594
Assay of human von Willebrand factor (2.7.21.).................. 268 Barium chloride dihydrate for homoeopathic
Assay of interferons (5.6.)........................................................ 689 preparations .......................................................................... 1594
Assay of pertussis vaccine (acellular) (2.7.16.)...................... 263 Barium sulfate ......................................................................... 1797
Assay of pertussis vaccine (whole cell) (2.7.7.) ..................... 252 Basic butylated methacrylate copolymer ...................... 9.4-5325
Assay of poliomyelitis vaccine (inactivated), in vivo BCG for immunotherapy......................................................... 894
(2.7.20.) .................................................................................... 267 BCG vaccine, freeze-dried....................................................... 895
Assay of tetanus vaccine (adsorbed) (2.7.8.) ......................... 253 Bearberry leaf.......................................................................... 1264
Assays (2.5.)............................................................................... 161 Beclometasone dipropionate ................................................. 1799
Astragalus mongholicus root ......................................... 9.2-4449 Beclometasone dipropionate monohydrate......................... 1802
Atenolol............................................................................. 9.1-4124 Beeswax, white ........................................................................ 1804
Atomic absorption spectrometry (2.2.23.) .............................. 37 Beeswax, yellow....................................................................... 1805
Atomic emission spectrometry (2.2.22.).................................. 36 Belamcanda chinensis rhizome............................................. 1266
Atomic emission spectrometry, inductively coupled plasma- Belladonna for homoeopathic preparations................. 9.2-4492
(2.2.57.) .................................................................................... 100 Belladonna leaf................................................................. 9.2-4452
Atomoxetine hydrochloride .................................................. 1767 Belladonna leaf dry extract, standardised .................... 9.2-4453
Atorvastatin calcium trihydrate............................................ 1769 Belladonna leaf tincture, standardised.......................... 9.2-4454
Atovaquone.............................................................................. 1771 Belladonna, prepared ...................................................... 9.2-4456
Atractylodes lancea rhizome ................................................. 1259 Benazepril hydrochloride ...................................................... 1806
Atractylodes rhizome, largehead .......................................... 1260 Bendroflumethiazide.............................................................. 1807
Atracurium besilate ................................................................ 1772 Benperidol ............................................................................... 1808
Atropine ............................................................................ 9.3-4847 Benserazide hydrochloride .................................................... 1810
Atropine sulfate................................................................ 9.3-4849 Bentonite.................................................................................. 1811
Aucklandia root ............................................................... 9.2-4450 Benzalkonium chloride.......................................................... 1811
Aujeszky’s disease vaccine (inactivated) for pigs ................ 1003 Benzalkonium chloride solution........................................... 1813
Aujeszky’s disease vaccine (live) for pigs for parenteral Benzathine benzylpenicillin .................................................. 1822
administration....................................................................... 1005 Benzbromarone....................................................................... 1815
Aurothiomalate, sodium ........................................................ 3565 Benzenesulfonate (methyl, ethyl and isopropyl) in active
Aurum chloratum natronatum for homoeopathic substances (2.5.41).................................................................. 180
preparations ................................................................... 9.5-5623 Benzethonium chloride ......................................................... 1816
Automatic rolling ball and falling ball viscometer methods Benzocaine........................................................................ 9.4-5326
(2.2.49.) ........................................................................... 9.3-4690 Benzoic acid ............................................................................ 1818
Avian infectious bronchitis vaccine (inactivated)............... 1007 Benzoin, Siam ......................................................................... 1272
Avian infectious bronchitis vaccine (live)............................ 1008 Benzoin, Sumatra.................................................................... 1273
Avian infectious bursal disease vaccine (inactivated) ........ 1010 Benzoin tincture, Siam........................................................... 1274
Avian infectious bursal disease vaccine (live) ..................... 1011 Benzoin tincture, Sumatra..................................................... 1274
Avian infectious encephalomyelitis vaccine (live) .............. 1013 Benzoyl peroxide, hydrous .................................................... 1819
Avian infectious laryngotracheitis vaccine (live)................ 1014 Benzyl alcohol .................................................................. 9.4-5327
Avian live virus vaccines : tests for extraneous agents in batches Benzyl benzoate ...................................................................... 1822
of finished product (2.6.25.).................................................. 222 Benzylpenicillin, benzathine ................................................. 1822
Avian paramyxovirus 1 (Newcastle disease) vaccine Benzylpenicillin potassium ............................................ 9.2-4505
(inactivated) .......................................................................... 1080 Benzylpenicillin, procaine .............................................. 9.2-4506
Avian paramyxovirus 1 (Newcastle disease) vaccine Benzylpenicillin sodium ................................................. 9.2-4508
(live) ....................................................................................... 1082 Betacarotene ............................................................................ 1829
Avian paramyxovirus 3 vaccine (inactivated) for turkeys.. 1016 Betacyclodextrin .............................................................. 9.3-4857
Avian tuberculin purified protein derivative....................... 3862 Betacyclodextrin, poly(hydroxypropyl) ether ..................... 2725
Avian viral tenosynovitis vaccine (live) ............................... 1017 Betadex.............................................................................. 9.3-4857
Avian viral vaccines : tests for extraneous agents in seed lots Betahistine dihydrochloride.................................................. 1831
(2.6.24.) .................................................................................... 219 Betahistine mesilate................................................................ 1832
Azaperone for veterinary use ................................................ 1778 Betamethasone ........................................................................ 1833
Azathioprine............................................................................ 1779 Betamethasone acetate ........................................................... 1835
Azelastine hydrochloride ....................................................... 1780 Betamethasone dipropionate.......................................... 9.3-4858
Azithromycin ................................................................... 9.3-4850 Betamethasone sodium phosphate....................................... 1839
Betamethasone valerate ......................................................... 1840
B Betaxolol hydrochloride......................................................... 1842
B19 virus (B19V), validation of nucleic acid amplification Bezafibrate ............................................................................... 1843
techniques for the quantification of B19V DNA in plasma Bicalutamide............................................................................ 1845
pools : guidelines..................................................................... 214 Bifonazole ................................................................................ 1846
Bacampicillin hydrochloride ................................................. 1787 Bilberry fruit, dried ................................................................ 1275
Bacitracin.......................................................................... 9.1-4129 Bilberry fruit dry extract, fresh, refined and standardised.. 1361
Bacitracin zinc.................................................................. 9.1-4133 Bilberry fruit, fresh................................................................. 1276
Baclofen ................................................................................... 1794 Biological assays (2.7.) ............................................................. 239
Bacterial endotoxins (2.6.14.)......................................... 9.3-4707 Biological assays and tests, statistical analysis of results of
Bacterial endotoxins, guidelines for using the test for (5.3.) ................................................................................ 9.2-4353
(5.1.10.) .................................................................................... 593 Biological indicators and related microbial preparations used
Bactericidal, fungicidal or yeasticidal activity of antiseptic in the manufacture of sterile products (5.1.2.) .......... 9.2-4336
medicinal products, determination of (5.1.11.) ......... 9.2-4348 Biological products, general texts on (5.2.) ........................... 599
Baical skullcap root ................................................................ 1262 Biological products, terminology used in monographs on
Bambuterol hydrochloride .................................................... 1795 (5.2.1.) ...................................................................................... 599
Barbados aloes ........................................................................ 1236 Biological tests (2.6.) ................................................................ 185
Barbary wolfberry fruit................................................... 9.3-4812 Biotin................................................................................. 9.5-5631
Barbital..................................................................................... 1797 Biperiden hydrochloride........................................................ 1848
Biphasic insulin injection ...................................................... 2770

Biphasic isophane insulin injection...................................... 2770 Bupivacaine hydrochloride.................................................... 1882

Birch leaf .................................................................................. 1276 Bupleurum root ............................................................... 9.4-5303
Bisacodyl.................................................................................. 1850 Buprenorphine ........................................................................ 1884
Bismuth subcarbonate............................................................ 1851 Buprenorphine hydrochloride .............................................. 1886
Bismuth subgallate.................................................................. 1852 Bursal disease vaccine (inactivated), infectious, avian....... 1010
Bismuth subnitrate, heavy ..................................................... 1853 Bursal disease vaccine (live), infectious, avian.................... 1011
Bismuth subsalicylate............................................................. 1854 Buserelin .................................................................................. 1887
Bisoprolol fumarate ................................................................ 1855 Buspirone hydrochloride ....................................................... 1889
Bistort rhizome ....................................................................... 1278 Busulfan ................................................................................... 1891
Bitter fennel ............................................................................. 1352 Butcher’s broom ...................................................................... 1296
Bitter-fennel fruit oil .............................................................. 1279 Butylated methacrylate copolymer, basic ..................... 9.4-5325
Bitter-fennel herb oil .............................................................. 1280 Butylhydroxyanisole ............................................................... 1893
Bitterness value (2.8.15.) .......................................................... 288 Butylhydroxytoluene .............................................................. 1894
Bitter-orange epicarp and mesocarp .................................... 1283 Butyl parahydroxybenzoate................................................... 1892
Bitter-orange-epicarp and mesocarp tincture..................... 1284
Bitter-orange flower................................................................ 1284 C
Bitter-orange-flower oil.......................................................... 1449 Cabergoline ............................................................................. 1897
Black cohosh............................................................................ 1285 Cachets.............................................................................. 9.4-5288
Blackcurrant leaf ..................................................................... 1290 Cadmium sulfate hydrate for homoeopathic prepara-
Black horehound..................................................................... 1289 tions........................................................................................ 1596
Bleomycin sulfate.................................................................... 1857 Cadmium sulfuricum for homoeopathic preparations...... 1596
Blood and blood components, empty sterile containers of Caffeine .................................................................................... 1898
plasticised poly(vinyl chloride) for (3.2.4.) ......................... 432 Caffeine monohydrate............................................................ 1899
Blood and blood components, human, materials for containers Calcifediol................................................................................ 1901
for (3.1.1.) ................................................................................ 391 Calcipotriol.............................................................................. 1902
Blood and blood components, sets for the transfusion of Calcipotriol monohydrate ..................................................... 1904
(3.2.6.) ...................................................................................... 433 Calcitonin (salmon)................................................................ 1906
Blood and blood components, sterile plastic containers for Calcitriol .................................................................................. 1909
(3.2.3.) ...................................................................................... 430 Calcium (2.4.3.)......................................................................... 132
Blood, anticoagulant and preservative solutions for .......... 1738 Calcium acetate....................................................................... 1910
Blood, sterile containers of plasticised poly(vinyl chloride) Calcium ascorbate .................................................................. 1911
containing anticoagulant solution (3.2.5.)........................... 432 Calcium carbonate.................................................................. 1912
Bogbean leaf ............................................................................ 1291 Calcium carboxymethylcellulose ................................... 9.4-5335
Boiling point (2.2.12.) ................................................................ 31 Calcium chloride dihydrate................................................... 1912
Boldo leaf................................................................................. 1292 Calcium chloride hexahydrate .............................................. 1913
Boldo leaf dry extract............................................................. 1293 Calcium dobesilate monohydrate ......................................... 1914
Borage (starflower) oil, refined ............................................. 1858 Calcium edetate, sodium ....................................................... 3568
Borax ........................................................................................ 1859 Calcium fluoratum for homoeopathic preparations ... 9.5-5624
Bordetella bronchiseptica vaccine (live) for dogs............... 1018 Calcium folinate...................................................................... 1914
Boric acid................................................................................. 1859 Calcium glucoheptonate ........................................................ 1917
Botulinum antitoxin ............................................................... 1115 Calcium gluconate .................................................................. 1917
Botulinum toxin type A for injection .................................. 1860 Calcium gluconate, anhydrous.............................................. 1918
Botulinum toxin type B for injection ................................... 1861 Calcium gluconate for injection ........................................... 1919
Bovine insulin .................................................................. 9.3-4929 Calcium glycerophosphate .................................................... 1920
Bovine leptospirosis vaccine (inactivated) .......................... 1019 Calcium hydrogen phosphate ............................................... 1921
Bovine parainfluenza virus vaccine (live)............................ 1021 Calcium hydrogen phosphate dihydrate.............................. 1922
Bovine respiratory syncytial virus vaccine (live) ................ 1022 Calcium hydroxide ................................................................. 1923
Bovine rhinotracheitis vaccine (inactivated), infectious ... 1067 Calcium in adsorbed vaccines (2.5.14.) ................................. 166
Bovine rhinotracheitis vaccine (live), infectious ................ 1068 Calcium iodatum for homoeopathic preparations ............. 1596
Bovine serum .......................................................................... 1863 Calcium iodide tetrahydrate for homoeopathic
Bovine tuberculin purified protein derivative..................... 3863 preparations .......................................................................... 1596
Bovine viral diarrhoea vaccine (inactivated) ...................... 1023 Calcium lactate........................................................................ 1923
Brimonidine tartrate .............................................................. 1864 Calcium lactate monohydrate ............................................... 1924
Bromazepam ........................................................................... 1865 Calcium lactate pentahydrate................................................ 1924
Bromhexine hydrochloride............................................. 9.1-4137 Calcium lactate trihydrate ..................................................... 1925
Bromocriptine mesilate.......................................................... 1868 Calcium levofolinate pentahydrate....................................... 1926
Bromperidol ............................................................................ 1870 Calcium levulinate dihydrate ................................................ 1928
Bromperidol decanoate.......................................................... 1872 Calcium pantothenate ............................................................ 1929
Brompheniramine maleate .................................................... 1873 Calcium pentetate (sodium) for radiopharmaceutical
Bronchitis vaccine (inactivated), infectious, avian ............. 1007 preparations ................................................................... 9.3-4800
Bronchitis vaccine (live), infectious, avian .......................... 1008 Calcium phosphate................................................................. 1929
Brotizolam ............................................................................... 1875 Calcium stearate ..................................................................... 1930
Brucellosis vaccine (live) (Brucella melitensis Rev. 1 strain) for Calcium sulfate dihydrate...................................................... 1932
veterinary use........................................................................ 1024 Calendula flower..................................................................... 1297
Buccal tablets and sublingual tablets............................. 9.3-4789 Calf coronavirus diarrhoea vaccine (inactivated)............... 1025
Buckwheat herb ...................................................................... 1294 Calf rotavirus diarrhoea vaccine (inactivated).................... 1026
Budesonide .............................................................................. 1876 Calicivirosis vaccine (inactivated), feline ............................ 1053
Bufexamac ............................................................................... 1878 Calicivirosis vaccine (live), feline ......................................... 1054
Buffer solutions (4.1.3.)................................................... 9.4-5252 Camphor, D-............................................................................ 1932
Buffer solutions (4.1.3.)................................................... 9.5-5549 Camphor, racemic .................................................................. 1934
Buflomedil hydrochloride...................................................... 1879 Candesartan cilexetil .............................................................. 1935
Bulk density and tapped density of powders (2.9.34.) ......... 359 Canine adenovirus vaccine (inactivated)............................. 1027
Bumetanide ............................................................................. 1881
Canine adenovirus vaccine (live).......................................... 1028 Catgut, sterile .......................................................................... 1207

Canine distemper vaccine (live) ........................................... 1029 Catgut, sterile, in distributor for veterinary use ................. 1219
Canine leptospirosis vaccine (inactivated) .......................... 1030 CD34/CD45+ cells in haematopoietic products, numeration
Canine parainfluenza virus vaccine (live) ........................... 1032 of (2.7.23.)................................................................................ 269
Canine parvovirosis vaccine (inactivated)........................... 1033 Cefaclor.................................................................................... 1969
Canine parvovirosis vaccine (live)........................................ 1034 Cefadroxil monohydrate........................................................ 1970
Cape aloes................................................................................ 1237 Cefalexin monohydrate.......................................................... 1972
Capecitabine............................................................................ 1937 Cefalotin sodium .................................................................... 1973
Cape jasmine fruit .................................................................. 1299 Cefamandole nafate................................................................ 1974
Capillary electrophoresis (2.2.47.) ............................................ 81 Cefapirin sodium.................................................................... 1976
Capillary viscometer method (2.2.9.)....................................... 27 Cefatrizine propylene glycol.................................................. 1977
Caprylate, sodium................................................................... 3569 Cefazolin sodium.................................................................... 1978
Caprylic acid ........................................................................... 1938 Cefepime dihydrochloride monohydrate ..................... 9.2-4513
Caprylocaproyl macrogolglycerides .............................. 9.1-4141 Cefixime............................................................................ 9.4-5335
Capsicum ................................................................................. 1300 Cefoperazone sodium ............................................................ 1984
Capsicum oleoresin, refined and standardised ................... 1302 Cefotaxime sodium ................................................................ 1985
Capsicum soft extract, standardised .................................... 1303 Cefoxitin sodium .................................................................... 1987
Capsicum tincture, standardised .......................................... 1304 Cefpodoxime proxetil ............................................................ 1989
Capsules ............................................................................ 9.4-5287 Cefprozil monohydrate ................................................... 9.4-5337
Capsules and tablets, disintegration of (2.9.1.) ..................... 299 Cefradine ................................................................................. 1994
Capsules, gastro-resistant ............................................... 9.4-5288 Ceftazidime pentahydrate............................................... 9.3-4865
Capsules, hard .................................................................. 9.4-5287 Ceftazidime pentahydrate with sodium carbonate for
Capsules, intrauterine .............................................................. 862 injection .......................................................................... 9.3-4867
Capsules, modified-release ............................................. 9.4-5288 Ceftriaxone sodium................................................................ 2000
Capsules, oromucosal...................................................... 9.3-4789 Cefuroxime axetil ............................................................ 9.2-4515
Capsules, rectal ......................................................................... 882 Cefuroxime sodium................................................................ 2003
Capsules, soft.................................................................... 9.4-5287 Celandine, greater................................................................... 1380
Capsules, vaginal....................................................................... 888 Celecoxib ................................................................................. 2004
Captopril .................................................................................. 1940 Celiprolol hydrochloride ....................................................... 2005
Caraway fruit........................................................................... 1305 Cell-based and gene therapy medicinal products, raw materials
Caraway oil .............................................................................. 1306 of biological origin for the production of (5.2.12.) ............ 627
Carbachol................................................................................. 1942 Cell-based preparations, microbiological examination of
Carbamazepine ....................................................................... 1943 (2.6.27.) ........................................................................... 9.2-4297
Carbasalate calcium................................................................ 1945 Cell count and viability, nucleated (2.7.29.).......................... 275
Carbidopa ......................................................................... 9.4-5333 Cell cultures for the production of veterinary vaccines
Carbimazole ............................................................................ 1948 (5.2.4.) ...................................................................................... 606
Carbocisteine........................................................................... 1949 Cell substrates for the production of vaccines for human use
Carbomers ............................................................................... 1950 (5.2.3.) ............................................................................. 9.3-4733
Carbon dioxide ....................................................................... 1951 Cellulose acetate .............................................................. 9.2-4516
Carbon dioxide in gases (2.5.24.) ........................................... 168 Cellulose acetate butyrate ...................................................... 2008
Carbon monoxide................................................................... 1953 Cellulose acetate phthalate .................................................... 2009
Carbon monoxide (15O) ......................................................... 1130 Cellulose, microcrystalline .................................................... 2010
Carbon monoxide in gases (2.5.25.)....................................... 169 Cellulose (microcrystalline) and carmellose sodium......... 3068
Carbon monoxide intermix (5 per cent) in nitrogen .. 9.3-4865 Cellulose, powdered ............................................................... 2013
Carboplatin.............................................................................. 1954 Centaury .................................................................................. 1310
Carboprost trometamol ......................................................... 1955 Centella .................................................................................... 1311
Carboxymethylcellulose......................................................... 1957 Cetirizine dihydrochloride .................................................... 2017
Carboxymethylcellulose calcium ................................... 9.4-5335 Cetostearyl alcohol ................................................................. 2019
Carboxymethylcellulose sodium........................................... 1958 Cetostearyl alcohol (type A), emulsifying ........................... 2019
Carboxymethylcellulose sodium, cross-linked ................... 2174 Cetostearyl alcohol (type B), emulsifying ........................... 2021
Carboxymethylcellulose sodium, low-substituted.............. 1958 Cetostearyl isononanoate ...................................................... 2022
Carisoprodol............................................................................ 1956 Cetostearyl sulfate, sodium ............................................ 9.4-5443
Carmellose............................................................................... 1957 Cetrimide................................................................................. 2022
Carmellose calcium ......................................................... 9.4-5335 Cetyl alcohol............................................................................ 2023
Carmellose sodium................................................................. 1958 Cetyl palmitate........................................................................ 2024
Carmellose sodium and microcrystalline cellulose............ 3068 Cetylpyridinium chloride ...................................................... 2025
Carmellose sodium, low-substituted.................................... 1958 Ceylon cinnamon bark oil ..................................................... 1318
Carmustine .............................................................................. 1960 Ceylon cinnamon leaf oil....................................................... 1318
Carnauba wax.......................................................................... 1960 CFC assay for human haematopoietic progenitor cells
Carprofen for veterinary use ................................................. 1961 (2.7.28.) .................................................................................... 274
Carrageenan ............................................................................ 1962 Chamomile flower, Roman ................................................... 1313
Carrier proteins for the production of conjugated Characterisation of crystalline and partially crystalline solids
polysaccharide vaccines for human use (5.2.11.) ............... 626 by X-ray powder diffraction (XRPD) (2.9.33.) ................... 354
Carteolol hydrochloride......................................................... 1963 Characterisation of crystalline solids by microcalorimetry and
Carvedilol ................................................................................ 1965 solution calorimetry (2.2.61.)................................................ 109
Cascara..................................................................................... 1307 Characterisation of preparations for nebulisation (2.9.44.).. 378
Cascara dry extract, standardised......................................... 1308 Characters section in monographs (5.11.) ............................ 729
Cassia oil .................................................................................. 1310 Charcoal, activated ................................................................. 2025
Castor oil, hydrogenated........................................................ 1966 Chemical imaging (5.24.) ............................................... 9.3-4767
Castor oil, polyoxyl................................................................. 2950 Chemical precursors for radiopharmaceutical
Castor oil, polyoxyl hydrogenated........................................ 2949 preparations ............................................................................ 813
Castor oil, refined ................................................................... 1967 Chemometric methods applied to analytical data (5.21.) ... 783
Castor oil, virgin ..................................................................... 1968 Chenodeoxycholic acid ......................................................... 2026

Chewable tablets .............................................................. 9.3-4792 Ciprofloxacin........................................................................... 2086

Chewing gums, medicated ............................................. 9.3-4784 Ciprofloxacin hydrochloride ................................................. 2088
Chewing gums, medicated, dissolution test for (2.9.25.)..... 340 Circular dichroism (2.2.41.) ...................................................... 69
Chicken anaemia vaccine (live), infectious .................. 9.5-5598 Cisatracurium besilate .................................................... 9.4-5347
Chicken flocks free from specified pathogens for the Cisplatin................................................................................... 2094
production and quality control of vaccines (5.2.2.) ........... 599 Citalopram hydrobromide.............................................. 9.3-4876
Chinese goldthread rhizome .......................................... 9.1-4102 Citalopram hydrochloride .............................................. 9.3-4877
Chitosan hydrochloride ......................................................... 2028 Citric acid ................................................................................ 2098
Chlamydiosis vaccine (inactivated), feline ......................... 1055 Citric acid monohydrate ........................................................ 2099
Chloral hydrate ....................................................................... 2029 Citronella oil............................................................................ 1319
Chlorambucil .......................................................................... 2029 Cladribine ................................................................................ 2100
Chloramine.............................................................................. 3816 Clarithromycin................................................................. 9.4-5351
Chloramphenicol ............................................................. 9.1-4142 Clarity and degree of opalescence of liquids (2.2.1.)... 9.2-4285
Chloramphenicol palmitate................................................... 2032 Clary sage oil ........................................................................... 1320
Chloramphenicol sodium succinate..................................... 2033 Classical swine-fever vaccine (live, prepared in cell
Chlorcyclizine hydrochloride................................................ 2034 cultures) ................................................................................. 1104
Chlordiazepoxide.................................................................... 2035 Clazuril for veterinary use..................................................... 2104
Chlordiazepoxide hydrochloride .......................................... 2036 Clebopride malate .................................................................. 2106
Chlorhexidine diacetate .................................................. 9.4-5339 Clemastine fumarate ....................................................... 9.3-4879
Chlorhexidine digluconate solution .............................. 9.3-4871 Clematis armandii stem .................................................. 9.3-4813
Chlorhexidine dihydrochloride ..................................... 9.4-5341 Clenbuterol hydrochloride .................................................... 2109
Chlorides (2.4.4.) ...................................................................... 132 Clindamycin hydrochloride ........................................... 9.3-4880
Chlormadinone acetate.......................................................... 2043 Clindamycin phosphate ......................................................... 2111
Chlorobutanol ......................................................................... 2045 Clioquinol................................................................................ 2114
Chlorobutanol hemihydrate .................................................. 2046 Clobazam ................................................................................. 2115
Chlorocresol ............................................................................ 2046 Clobetasol propionate ............................................................ 2116
Chloroquine phosphate ......................................................... 2047 Clobetasone butyrate.............................................................. 2118
Chloroquine sulfate ................................................................ 2048 Clodronate disodium tetrahydrate ....................................... 2119
Chlorphenamine maleate ...................................................... 2048 Clofazimine ............................................................................. 2120
Chlorpromazine hydrochloride ............................................ 2050 Clofibrate ................................................................................. 2122
Chlorpropamide ..................................................................... 2051 Clomifene citrate ............................................................. 9.5-5635
Chlorprothixene hydrochloride............................................ 2052 Clomipramine hydrochloride ............................................... 2124
Chlortalidone .......................................................................... 2054 Clonazepam............................................................................. 2126
Chlortetracycline hydrochloride........................................... 2055 Clonidine hydrochloride........................................................ 2127
Cholecalciferol ........................................................................ 2058 Clopamide ............................................................................... 2128
Cholecalciferol concentrate (oily form)............................... 2059 Clopidogrel besilate................................................................ 2129
Cholecalciferol concentrate (powder form) ........................ 2061 Clopidogrel hydrochloride .................................................... 2131
Cholecalciferol concentrate (water-dispersible form)........ 2062 Clopidogrel hydrogen sulfate ................................................ 2133
Cholera vaccine (inactivated), fowl ...................................... 1063 Clorazepate, dipotassium....................................................... 2291
Cholera vaccine (inactivated, oral)......................................... 898 Closantel sodium dihydrate for veterinary use................... 2134
Cholesterol............................................................................... 2064 Clostridium botulinum vaccine for veterinary use ............ 1035
Cholesterol for parenteral use ........................................ 9.2-4517 Clostridium chauvoei vaccine for veterinary use ............... 1036
Cholesterol in oils rich in omega-3 acids, total (2.4.32.) ..... 157 Clostridium novyi (type B) vaccine for veterinary use...... 1036
Choline ([11C]methyl) injection..................................... 9.4-5297 Clostridium perfringens vaccine for veterinary use........... 1038
Chondroitin sulfate sodium .................................................. 2067 Clostridium septicum vaccine for veterinary use............... 1040
Chromatographic separation techniques (2.2.46.) ...... 9.2-4286 Closures and containers for parenteral preparations
Chromatography, gas (2.2.28.).................................................. 44 and ophthalmic preparations, polypropylene for
Chromatography, liquid (2.2.29.) ............................................. 46 (3.1.6.) ............................................................................. 9.4-5123
Chromatography, paper (2.2.26.) ............................................. 42 Closures and containers for pharmaceutical use, plastic
Chromatography, size-exclusion (2.2.30.) ............................... 47 (3.2.2.) ...................................................................................... 428
Chromatography, supercritical fluid (2.2.45.)......................... 74 Closures and tubing, silicone elastomer for (3.1.9.)............. 409
Chromatography, thin-layer (2.2.27.) ...................................... 43 Closures for containers for aqueous parenteral preparations,
Chromium (51Cr) edetate injection ...................................... 1131 for powders and for freeze-dried powders, rubber
Chymotrypsin ......................................................................... 2069 (3.2.9.) ............................................................................. 9.5-5545
Ciclesonide .............................................................................. 2070 Clotrimazole............................................................................ 2136
Ciclopirox ................................................................................ 2071 Clove......................................................................................... 1322
Ciclopirox olamine ................................................................. 2072 Clove oil ................................................................................... 1323
Ciclosporin .............................................................................. 2074 Cloxacillin sodium ................................................................. 2137
Cilastatin sodium.................................................................... 2075 Clozapine ................................................................................. 2139
Cilazapril.................................................................................. 2077 Coagulation factor II, human, assay of (2.7.18.)................... 266
Cimetidine ........................................................................ 9.4-5343 Coagulation factor IX, human .............................................. 2674
Cimetidine hydrochloride .............................................. 9.4-5345 Coagulation factor IX, human, assay of (2.7.11.) ................. 259
Cinchocaine hydrochloride ................................................... 2082 Coagulation factor IX (rDNA) concentrated solution,
Cinchona bark......................................................................... 1314 human ............................................................................. 9.3-4915
Cinchona liquid extract, standardised ................................. 1316 Coagulation factor IX (rDNA) powder for solution for
Cineole ..................................................................................... 2083 injection, human............................................................ 9.3-4920
Cineole in essential oils, 1,8-, assay of (2.8.11.) .................... 285 Coagulation factors, activated (2.6.22.) ................................. 219
Cineole type niaouli oil.......................................................... 1453 Coagulation factors, assay of heparin (2.7.12.) ..................... 259
Cinnamon................................................................................ 1317 Coagulation factor VIIa (rDNA) concentrated solution,
Cinnamon bark oil, Ceylon................................................... 1318 human ............................................................................. 9.5-5687
Cinnamon leaf oil, Ceylon..................................................... 1318 Coagulation factor VII, human ............................................ 2666
Cinnarizine.............................................................................. 2084 Coagulation factor VII, human, assay of (2.7.10.)................ 258
Ciprofibrate ............................................................................. 2085 Coagulation factor VIII, human ........................................... 2672
Coagulation factor VIII, human, assay of (2.7.4.) ................ 246 Containers for aqueous solutions for intravenous infusion,
Coagulation factor VIII (rDNA), human ............................ 2673 materials based on plasticised poly(vinyl chloride) for
Coagulation factor X, assay of (2.7.19.) ................................. 266 (3.1.14.) .................................................................................... 416
Coagulation factor XI, human .............................................. 2681 Containers for human blood and blood components, materials
Coagulation factor XI, human, assay of (2.7.22.) ................. 269 based on plasticised poly(vinyl chloride) for (3.1.1.1.)...... 391
Coated granules ........................................................................ 860 Containers for human blood and blood components, materials
Coated tablets................................................................... 9.3-4791 for (3.1.1.) ................................................................................ 391
Cocaine hydrochloride........................................................... 2140 Containers for human blood and blood components, plastic,
Coccidiosis vaccine (live) for chickens ................................ 1042 sterile (3.2.3.)........................................................................... 430
Cocculus for homoeopathic preparations ........................... 1597 Containers for non-injectable aqueous solutions, materials
Coconut oil, refined................................................................ 2141 based on non-plasticised poly(vinyl chloride) for
Cocoyl caprylocaprate............................................................ 2142 (3.1.10.) .................................................................................... 410
Codeine hydrochloride dihydrate.................................. 9.5-5636 Containers for parenteral preparations and for
Codeine monohydrate..................................................... 9.5-5639 ophthalmic preparations, polyethylene with additives for
Codeine phosphate hemihydrate ................................... 9.5-5641 (3.1.5.) ............................................................................. 9.4-5120
Codeine phosphate sesquihydrate ........................................ 2148 Containers for parenteral preparations and for ophthalmic
Codergocrine mesilate ........................................................... 2149 preparations, polyethylene without additives for
Cod-liver oil, farmed.............................................................. 2151 (3.1.4.) ............................................................................. 9.2-4310
Cod-liver oil (type A)............................................................. 2155 Containers for pharmaceutical use, glass (3.2.1.) ................. 423
Cod-liver oil (type B) ............................................................. 2159 Containers for preparations not for parenteral use,
Codonopsis root .............................................................. 9.3-4815 polyethylene terephthalate for (3.1.15) ................................ 419
Coix seed........................................................................... 9.2-4457 Containers for solid dosage forms for oral administration,
Cola .......................................................................................... 1326 materials based on non-plasticised poly(vinyl chloride) for
Colchicine......................................................................... 9.4-5354 (3.1.11.) .................................................................................... 412
Cold-water vibriosis vaccine (inactivated) for Containers, materials used for the manufacture of (3.1.).... 391
salmonids........................................................................ 9.2-4428 Containers of plasticised poly(vinyl chloride) for human blood
Colestyramine ......................................................................... 2164 and blood components, empty sterile (3.2.4.)..................... 432
Colibacillosis vaccine (inactivated), neonatal piglet .......... 1077 Containers of plasticised poly(vinyl chloride) for human blood
Colibacillosis vaccine (inactivated), neonatal ruminant.... 1079 containing anticoagulant solution, sterile (3.2.5.) .............. 432
Colistimethate sodium.................................................... 9.4-5356 Contamination, microbial : microbial enumeration tests
Colistin sulfate ........................................................................ 2166 (2.6.12.) .................................................................................... 195
Colloidal anhydrous silica ..................................................... 3549 Contamination, microbial : test for specified micro-organisms
Colloidal hydrated silica ........................................................ 3550 (2.6.13.) .................................................................................... 199
Colloidal silica, hydrophobic ................................................ 3551 Content uniformity of single-dose preparations (2.9.6.) ..... 312
Colloidal silver, for external use ........................................... 3552 Control of impurities in substances for pharmaceutical use
Colony-forming cell assay for human haematopoietic (5.10.) ....................................................................................... 723
progenitor cells (2.7.28.) ........................................................ 274 Control of microbiological quality, alternative methods for
Colophony ............................................................................... 1327 (5.1.6.) ............................................................................. 9.2-4339
Coloration of liquids (2.2.2.) ..................................................... 22 Copolymer, basic butylated methacrylate .................... 9.4-5325
Common selfheal fruit-spike ................................................ 1327 Copolymer, grafted, macrogol poly(vinyl alcohol) ............ 2945
Common stinging nettle for homoeopathic preparations.. 1614 Copolymer, methacrylic acid - ethyl acrylate (1:1) ............ 3015
Comparative table of porosity of sintered-glass filters Copolymer, methacrylic acid - ethyl acrylate (1:1) dispersion
(2.1.2.) ........................................................................................ 15 30 per cent ............................................................................. 3017
Complexometric titrations (2.5.11.) ....................................... 164 Copolymer, methacrylic acid – methyl methacrylate
Composition of fatty acids by gas chromatography (1:1) ........................................................................................ 3018
(2.4.22.) .................................................................................... 141 Copolymer, methacrylic acid - methyl methacrylate
Composition of fatty acids in oils rich in omega-3 acids (1:2) ........................................................................................ 3019
(2.4.29.) .................................................................................... 155 Copolymer (type A), ammonio methacrylate..................... 1712
Compressed lozenges ...................................................... 9.3-4789 Copolymer (type B), ammonio methacrylate ..................... 1713
Concentrated solutions for haemodialysis .......................... 2628 Copovidone ...................................................................... 9.3-4882
Concentrates for injections or infusions................................ 872 Copper acetate monohydrate for homoeopathic
Concentrates for intrauterine solutions ................................. 862 preparations .......................................................................... 1599
Conductivity (2.2.38.) ................................................................ 62 Copper for homoeopathic preparations .............................. 1600
Coneflower herb, purple ........................................................ 1486 Copper sulfate ......................................................................... 2169
Coneflower root, narrow-leaved ........................................... 1447 Copper sulfate pentahydrate ................................................. 2170
Coneflower root, pale............................................................. 1467 Copper tetramibi tetrafluoroborate for radiopharmaceutical
Coneflower root, purple......................................................... 1488 preparations ................................................................... 9.2-4435
Conjugated estrogens ............................................................. 2410 Coriander................................................................................. 1329
Conjugated polysaccharide vaccines for human use, carrier Coriander oil ........................................................................... 1330
proteins for the production of (5.2.11.) ............................... 626 Coronavirus diarrhoea vaccine (inactivated), calf ............. 1025
Consistency by penetrometry, measurement of (2.9.9.)...... 313 Cortisone acetate .................................................................... 2170
Containers (3.2.) ....................................................................... 423 Cotton, absorbent ................................................................... 2172
Containers and closures for parenteral preparations Cottonseed oil, hydrogenated .............................................. 2173
and ophthalmic preparations, polypropylene for Couch grass rhizome.............................................................. 1331
(3.1.6.) ............................................................................. 9.4-5123 Creams ....................................................................................... 883
Containers and closures for pharmaceutical use, plastic Cresol, crude ........................................................................... 2173
(3.2.2.) ...................................................................................... 428 Crocus for homoeopathic preparations ............................... 1599
Containers and tubing for total parenteral nutrition Cromoglicate, sodium............................................................ 3574
preparations, poly(ethylene - vinyl acetate) for Croscarmellose sodium ......................................................... 2174
(3.1.7.) ............................................................................. 9.2-4318 Crospovidone .......................................................................... 2175
Containers for aqueous solutions for infusion, plastic Crotamiton .............................................................................. 2177
(3.2.2.1.) ................................................................................... 429 Crystalline and partially crystalline solids, characterisation
by X-ray powder diffraction (XRPD) of (2.9.33.)............... 354

Crystalline solids, characterisation by microcalorimetry and Detomidine hydrochloride for veterinary use .................... 2217
solution calorimetry (2.2.61.)................................................ 109 Devil’s claw dry extract .......................................................... 1334
Crystallinity (5.16.)................................................................... 757 Devil’s claw root...................................................................... 1335
Cuprum aceticum for homoeopathic preparations............ 1599 Dexamethasone....................................................................... 2218
Cuprum metallicum for homoeopathic preparations........ 1600 Dexamethasone acetate.......................................................... 2220
Cutaneous application, liquid preparations for .................... 864 Dexamethasone isonicotinate ............................................... 2222
Cutaneous application, powders for....................................... 874 Dexamethasone sodium phosphate...................................... 2223
Cutaneous application, semi-solid preparations for ............ 882 Dexchlorpheniramine maleate.............................................. 2226
Cutaneous application, veterinary liquid preparations Dexpanthenol.......................................................................... 2227
for .................................................................................... 9.3-4792 Dextran 1 for injection.................................................... 9.4-5363
Cutaneous foams ...................................................................... 864 Dextran 40 for injection ........................................................ 2229
Cutaneous patches .................................................................... 882 Dextran 60 for injection ........................................................ 2230
Cyanocobalamin ..................................................................... 2178 Dextran 70 for injection ........................................................ 2231
Cyanocobalamin (57Co) capsules .......................................... 1132 Dextranomer ........................................................................... 2231
Cyanocobalamin (57Co) solution .......................................... 1133 Dextrans, molecular mass distribution in (2.2.39.)................ 62
Cyanocobalamin (58Co) capsules .......................................... 1134 Dextrin .............................................................................. 9.2-4527
Cyanocobalamin (58Co) solution .......................................... 1134 Dextromethorphan hydrobromide....................................... 2233
Cyclamate, sodium ................................................................. 3576 Dextromoramide tartrate ...................................................... 2234
Cyclizine hydrochloride......................................................... 2179 Dextropropoxyphene hydrochloride.................................... 2235
Cyclopentolate hydrochloride ............................................... 2180 Dextrose............................................................................ 9.1-4167
Cyclophosphamide ................................................................. 2181 Dextrose monohydrate.................................................... 9.1-4168
Cyproheptadine hydrochloride............................................. 2182 Diacerein.................................................................................. 2236
Cyproterone acetate................................................................ 2183 Diazepam................................................................................. 2238
Cysteine hydrochloride monohydrate.................................. 2185 Diazoxide ................................................................................. 2240
Cystine ..................................................................................... 2187 Dibrompropamidine diisetionate ......................................... 2240
Cytarabine ............................................................................... 2188 Dibutyl phthalate .................................................................... 2241
Dichlorobenzyl alcohol, 2,4- ................................................. 2242
D Dichloromethane .................................................................... 3034
Dacarbazine............................................................................. 2193 Diclazuril for veterinary use.................................................. 2243
Dalteparin sodium.................................................................. 2194 Diclofenac potassium ............................................................. 2245
Danaparoid sodium................................................................ 2196 Diclofenac sodium.................................................................. 2246
Dandelion herb with root ...................................................... 1332 Dicloxacillin sodium .............................................................. 2248
Dandelion root........................................................................ 1333 Dicycloverine hydrochloride.......................................... 9.1-4147
Dapsone ................................................................................... 2198 Didanosine .............................................................................. 2250
Daunorubicin hydrochloride ................................................ 2199 Dienogest ................................................................................. 2252
D-Camphor .............................................................................. 1932 Diethylcarbamazine citrate.................................................... 2254
Decyl oleate ............................................................................. 2200 Diethylene glycol and ethylene glycol in ethoxylated
Deferiprone ...................................................................... 9.5-5647 substances (2.4.30.)................................................................. 157
Deferoxamine mesilate.................................................... 9.3-4887 Diethylene glycol monoethyl ether ...................................... 2256
Degree of coloration of liquids (2.2.2.) .................................... 22 Diethylene glycol palmitostearate......................................... 2257
Dembrexine hydrochloride monohydrate for veterinary Diethyl phthalate .................................................................... 2253
use.................................................................................... 9.2-4526 Diethylstilbestrol..................................................................... 2258
Demeclocycline hydrochloride ............................................. 2203 Diffraction, laser light, particle size analysis by (2.9.31.) ... 349
Demonstration of uniformity of dosage units using large Difloxacin hydrochloride trihydrate for veterinary use..... 2259
sample sizes (2.9.47.).............................................................. 384 Digitalis leaf............................................................................. 1336
Density of powders, bulk density and tapped (2.9.34.)........ 359 Digitoxin .................................................................................. 2260
Density of solids (2.2.42.) .......................................................... 70 Digoxin .................................................................................... 2261
Density of solids, gas pycnometric (2.9.23.).......................... 339 Dihydralazine sulfate, hydrated ..................................... 9.2-4528
Density, relative (2.2.5.) ............................................................. 25 Dihydrocodeine hydrogen tartrate ....................................... 2266
Dental type silica .................................................................... 3550 Dihydroergocristine mesilate ................................................ 2267
Depressor substances (2.6.11.) ................................................ 195 Dihydroergotamine mesilate................................................. 2269
Deptropine citrate................................................................... 2205 Dihydroergotamine tartrate .................................................. 2271
Dequalinium chloride ............................................................ 2206 Dihydrostreptomycin sulfate for veterinary use ................. 2272
Desacyl-4-monophosphoryl lipid A, 3-O- ......................... 2207 Dihydrotachysterol ................................................................. 2274
Desflurane................................................................................ 2209 Diltiazem hydrochloride........................................................ 2276
Desipramine hydrochloride................................................... 2210 Dimenhydrinate...................................................................... 2277
Deslanoside ............................................................................. 2211 Dimercaprol ............................................................................ 2279
Desloratadine .......................................................................... 2212 Dimethylacetamide ................................................................ 2280
Desmopressin .......................................................................... 2213 Dimethylaniline, N,N- (2.4.26.) .............................................. 152
Desogestrel .............................................................................. 2215 Dimethyl sulfoxide ................................................................. 2279
Desoxycortone acetate ........................................................... 2216 Dimeticone .............................................................................. 2281
Detection and measurement of radioactivity (2.2.66.) ........ 113 Dimetindene maleate ............................................................. 2282
Detector tubes, gas (2.1.6.) ............................................. 9.3-4685 Dinoprostone ................................................................... 9.2-4529
Determination of aflatoxin B1 in herbal drugs (2.8.18.) ...... 289 Dinoprost trometamol ........................................................... 2283
Determination of bactericidal, fungicidal or yeasticidal activity Dioscorea nipponica rhizome ........................................ 9.5-5613
of antiseptic medicinal products (5.1.11.) .................. 9.2-4348 Dioscorea oppositifolia rhizome........................................... 1337
Determination of elemental impurities (2.4.20.) ......... 9.5-5539 Diosmin ................................................................................... 2286
Determination of nitrogen by sulfuric acid digestion Dioxan and ethylene oxide (2.4.25.)....................................... 151
(2.5.9.) ...................................................................................... 164 Dip concentrates .............................................................. 9.3-4792
Determination of primary aromatic amino-nitrogen Diphenhydramine hydrochloride ......................................... 2288
(2.5.8.) ...................................................................................... 163 Diphenoxylate hydrochloride................................................ 2289
Determination of water by distillation (2.2.13.) ..................... 31 Diphtheria and tetanus toxins and toxoids, flocculation value
(Lf) of, (Ramon assay) (2.7.27.)............................................ 273
Diphtheria and tetanus vaccine (adsorbed) .......................... 899 Docetaxel trihydrate ............................................................... 2307
Diphtheria and tetanus vaccine (adsorbed, reduced antigen(s) Docusate sodium .................................................................... 2309
content).................................................................................... 900 Dodecyl gallate........................................................................ 2309
Diphtheria antitoxin............................................................... 1115 Dog rose................................................................................... 1338
Diphtheria, tetanus and hepatitis B (rDNA) vaccine Domperidone .......................................................................... 2310
(adsorbed) ............................................................................... 901 Domperidone maleate............................................................ 2312
Diphtheria, tetanus and pertussis (acellular, component) Dopamine hydrochloride ...................................................... 2313
vaccine (adsorbed) ................................................................. 902 Dopexamine dihydrochloride ............................................... 2315
Diphtheria, tetanus and pertussis (acellular, component) Dorzolamide hydrochloride .................................................. 2316
vaccine (adsorbed, reduced antigen(s) content)................. 903 Dosage forms (glossary) ................................................. 9.2-4423
Diphtheria, tetanus and pertussis (whole cell) vaccine Dosage units, demonstration of uniformity using large sample
(adsorbed) ............................................................................... 905 sizes (2.9.47.) ........................................................................... 384
Diphtheria, tetanus and poliomyelitis (inactivated) vaccine Dosage units, uniformity of (2.9.40.) ............................ 9.1-4055
(adsorbed, reduced antigen(s) content)............................... 906 Dosulepin hydrochloride................................................ 9.4-5364
Diphtheria, tetanus, pertussis (acellular, component) and DOTATOC (gallium (68Ga)) injection ................................. 1150
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5583 Doxapram hydrochloride ...................................................... 2319
Diphtheria, tetanus, pertussis (acellular, component) and Doxazosin mesilate................................................................. 2320
hepatitis B (rDNA) vaccine (adsorbed) ............................... 910 Doxepin hydrochloride .......................................................... 2322
Diphtheria, tetanus, pertussis (acellular, component) and Doxorubicin hydrochloride................................................... 2323
poliomyelitis (inactivated) vaccine (adsorbed)................... 911 Doxycycline hyclate ................................................................ 2324
Diphtheria, tetanus, pertussis (acellular, component) and Doxycycline monohydrate..................................................... 2326
poliomyelitis (inactivated) vaccine (adsorbed, reduced Doxylamine hydrogen succinate........................................... 2328
antigen(s) content) ................................................................. 913 Droperidol ............................................................................... 2329
Diphtheria, tetanus, pertussis (acellular, component), Droppers (2.1.1.)......................................................................... 15
hepatitis B (rDNA), poliomyelitis (inactivated) and Drop point (2.2.17.).................................................................... 33
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5585 Drops (nasal) and sprays (liquid nasal) ................................. 867
Diphtheria, tetanus, pertussis (acellular, component), Drops, oral........................................................................ 9.3-4786
poliomyelitis (inactivated) and haemophilus type b conjugate Drospirenone .......................................................................... 2331
vaccine (adsorbed) ........................................................ 9.5-5587 Dry extracts ............................................................................... 818
Diphtheria, tetanus, pertussis (whole cell) and poliomyelitis Drynaria rhizome ............................................................ 9.2-4459
(inactivated) vaccine (adsorbed) .......................................... 920 Dry residue of extracts (2.8.16.).............................................. 289
Diphtheria, tetanus, pertussis (whole cell), poliomyelitis Duck plague vaccine (live)..................................................... 1046
(inactivated) and haemophilus type b conjugate vaccine Duck viral hepatitis type I vaccine (live) ............................. 1047
(adsorbed) ...................................................................... 9.5-5590 Duloxetine hydrochloride...................................................... 2332
Diphtheria vaccine (adsorbed)................................................ 924 Dutasteride .............................................................................. 2334
Diphtheria vaccine (adsorbed), assay of (2.7.6.)................... 247 Dwarf pine oil ......................................................................... 1340
Diphtheria vaccine (adsorbed, reduced antigen content) ... 925 Dydrogesterone....................................................................... 2336
Dipivefrine hydrochloride ..................................................... 2290
Dipotassium clorazepate........................................................ 2291 E
Dipotassium phosphate ......................................................... 2292 Ear drops and ear sprays.......................................................... 856
Diprophylline .......................................................................... 2293 Ear powders............................................................................... 856
Dipyridamole .......................................................................... 2294 Ear preparations........................................................................ 855
Dirithromycin ......................................................................... 2296 Ear preparations, semi-solid ................................................... 856
Disintegration of suppositories and pessaries (2.9.2.) ......... 301 Ear sprays and ear drops.......................................................... 856
Disintegration of tablets and capsules (2.9.1.) ...................... 299 Ear tampons .............................................................................. 856
Disodium clodronate tetrahydrate ....................................... 2119 Ear washes ................................................................................. 856
Disodium edetate.................................................................... 2297 Ebastine.................................................................................... 2341
Disodium etidronate .............................................................. 2433 Eclipta herb ...................................................................... 9.2-4461
Disodium pamidronate pentahydrate .................................. 3259 Econazole................................................................................. 2342
Disodium phosphate .............................................................. 2298 Econazole nitrate .................................................................... 2343
Disodium phosphate dihydrate............................................. 2299 Edetate (chromium (51Cr)) injection.................................... 1131
Disodium phosphate dodecahydrate.................................... 2299 Edetate, disodium ................................................................... 2297
Disopyramide.......................................................................... 2300 Edetate, sodium calcium........................................................ 3568
Disopyramide phosphate ....................................................... 2301 Edetic acid ............................................................................... 2344
Dispersible tablets............................................................ 9.3-4792 Edotreotide (gallium (68Ga)) injection................................. 1150
Dissolution, apparent (2.9.43.)................................................ 377 Edrophonium chloride........................................................... 2345
Dissolution, intrinsic (2.9.29.) ................................................ 347 Effervescent granules ............................................................... 860
Dissolution test for lipophilic solid dosage forms (2.9.42.).. 377 Effervescent powders ............................................................... 875
Dissolution test for solid dosage forms (2.9.3.) .................... 302 Effervescent tablets.......................................................... 9.3-4791
Dissolution test for transdermal patches (2.9.4.).................. 309 Efficacy of antimicrobial preservation (5.1.3.)...................... 577
Dissolution testing, recommendations on (5.17.1.) ............. 761 Efficacy of veterinary vaccines and immunosera, evaluation of
Distemper vaccine (live), canine........................................... 1029 (5.2.7.) ...................................................................................... 612
Distemper vaccine (live) for mustelids ................................ 1045 Egg drop syndrome 76 vaccine (inactivated) ..................... 1048
Distillation range (2.2.11.)......................................................... 30 Egg phospholipids for injection..................................... 9.2-4533
Distribution estimation by analytical sieving, particle-size Elder flower ............................................................................. 1342
(2.9.38.) .................................................................................... 367 Electrophoresis (2.2.31.) ............................................................ 48
Disulfiram................................................................................ 2302 Electrophoresis, capillary (2.2.47.) ........................................... 81
Dithranol ................................................................................. 2303 Elemental impurities (5.20.)........................................... 9.3-4759
DL-Methionine ........................................................................ 3025 Elemental impurities, determination of (2.4.20.) ........ 9.5-5539
DL-α-Tocopheryl hydrogen succinate................................... 3806 Eleutherococcus...................................................................... 1344
Dobesilate monohydrate, calcium ........................................ 1914 Emedastine difumarate .......................................................... 2346
Dobutamine hydrochloride ................................................... 2304 Emetine hydrochloride pentahydrate .................................. 2347
Docetaxel ................................................................................. 2305

Empty sterile containers of plasticised poly(vinyl chloride) for Ethanol, anhydrous................................................................. 2419
human blood and blood components (3.2.4.)..................... 432 Ethanol content (2.9.10.) ......................................................... 315
Emulsifying cetostearyl alcohol (type A)............................. 2019 Ether ......................................................................................... 2421
Emulsifying cetostearyl alcohol (type B) ............................. 2021 Ether, anaesthetic.................................................................... 2422
Emulsions, solutions and suspensions, oral ................. 9.3-4786 Ethinylestradiol ....................................................................... 2422
Enalaprilat dihydrate.............................................................. 2349 Ethionamide ............................................................................ 2424
Enalapril maleate .................................................................... 2348 Ethosuximide ................................................................... 9.4-5369
Encephalitis vaccine (inactivated), tick-borne...................... 988 Ethoxylated substances, ethylene glycol and diethylene glycol
Encephalomyelitis vaccine (live), infectious, avian ............ 1013 in (2.4.30.) ............................................................................... 157
Endotoxins, bacterial (2.6.14.) ....................................... 9.3-4707 Ethyl acetate ............................................................................ 2427
Endotoxins, bacterial, guidelines for using the test for Ethyl acrylate - methacrylic acid copolymer (1:1).............. 3015
(5.1.10.) .................................................................................... 593 Ethyl acrylate - methacrylic acid copolymer (1:1) dispersion
Enilconazole for veterinary use ............................................ 2351 30 per cent ............................................................................. 3017
Enoxaparin sodium ................................................................ 2352 Ethylcellulose.................................................................... 9.2-4539
Enoxolone ................................................................................ 2354 Ethylenediamine ..................................................................... 2431
Enrofloxacin for veterinary use...................................... 9.2-4535 Ethylene glycol and diethylene glycol in ethoxylated substances
Entacapone .............................................................................. 2357 (2.4.30.) .................................................................................... 157
Entecavir monohydrate.......................................................... 2359 Ethylene glycol monopalmitostearate .................................. 2430
Enzootic pneumonia vaccine (inactivated), porcine.......... 1086 Ethylene glycol monostearate................................................ 2430
Ephedra herb ........................................................................... 1346 Ethylene oxide and dioxan (2.4.25.) ....................................... 151
Ephedrine ................................................................................ 2360 Ethylhexanoic acid, 2- (2.4.28.)............................................... 154
Ephedrine hemihydrate ......................................................... 2361 Ethylmorphine hydrochloride............................................... 2431
Ephedrine hydrochloride....................................................... 2362 Ethyl oleate .............................................................................. 2428
Ephedrine hydrochloride, racemic ....................................... 2363 Ethyl parahydroxybenzoate ................................................... 2428
Epinastine hydrochloride....................................................... 2364 Ethyl parahydroxybenzoate sodium..................................... 3577
Epinephrine ............................................................................. 1650 Etidronate disodium............................................................... 2433
Epinephrine tartrate ............................................................... 1652 Etilefrine hydrochloride......................................................... 2433
Epirubicin hydrochloride ...................................................... 2365 Etodolac ................................................................................... 2435
Eplerenone............................................................................... 2367 Etofenamate............................................................................. 2437
Eptacog alfa (activated) concentrated solution............ 9.5-5687 Etomidate................................................................................. 2439
Equine herpesvirus vaccine (inactivated)............................ 1050 Etoposide .......................................................................... 9.1-4152
Equine influenza vaccine (inactivated) ................................ 1051 Eucalyptus leaf ........................................................................ 1349
Equisetum stem ...................................................................... 1347 Eucalyptus oil .......................................................................... 1349
Ergocalciferol .......................................................................... 2368 Eucommia bark................................................................ 9.2-4462
Ergoloid mesilates................................................................... 2149 Eugenol .................................................................................... 2443
Ergometrine maleate .............................................................. 2370 European goldenrod............................................................... 1376
Ergotamine tartrate ................................................................ 2371 European viper venom antiserum ........................................ 1119
Erysipelas vaccine (inactivated), swine................................ 1104 Evaluation of efficacy of veterinary vaccines and immunosera
Erythritol ................................................................................. 2373 (5.2.7.) ...................................................................................... 612
Erythromycin .......................................................................... 2374 Evaluation of safety of each batch of immunosera for
Erythromycin estolate ............................................................ 2378 veterinary use (5.2.9.)............................................................. 625
Erythromycin ethylsuccinate.......................................... 9.3-4893 Evaluation of safety of veterinary vaccines and immunosera
Erythromycin lactobionate ............................................. 9.2-4536 (5.2.6.) ...................................................................................... 610
Erythromycin stearate............................................................ 2388 Evening primrose oil, refined................................................ 2444
Erythropoietin concentrated solution.................................. 2391 Evodia fruit....................................................................... 9.2-4464
Escitalopram............................................................................ 2395 Excipients, functionality-related characteristics of
Escitalopram oxalate .............................................................. 2397 (5.15.) .............................................................................. 9.2-4411
Eserine salicylate..................................................................... 3335 Exemestane.............................................................................. 2445
Esketamine hydrochloride.............................................. 9.3-4895 Extemporaneous preparation of radiopharmaceuticals
Esomeprazole magnesium dihydrate ................................... 2401 (5.19.) ....................................................................................... 767
Esomeprazole magnesium trihydrate................................... 2402 Extractable volume of parenteral preparations, test for
Essential oils .............................................................................. 814 (2.9.17.) .................................................................................... 322
Essential oils, assay of 1,8-cineole in (2.8.11.)....................... 285 Extracts, dry .............................................................................. 818
Essential oils, fatty oils and resinified essential oils in Extracts, dry residue of (2.8.16.)............................................. 289
(2.8.7.) ...................................................................................... 284 Extracts, herbal drug ................................................................ 815
Essential oils, foreign esters in (2.8.6.)................................... 284 Extracts, herbal drug, monographs on (information chapter)
Essential oils in herbal drugs (2.8.12.) ................................... 285 (5.23.) ....................................................................................... 807
Essential oils, odour and taste (2.8.8.) ................................... 284 Extracts, liquid (fluid) .............................................................. 817
Essential oils, residue on evaporation (2.8.9.)....................... 284 Extracts, loss on drying of (2.8.17.)........................................ 289
Essential oils, solubility in alcohol (2.8.10.) .......................... 284 Extracts, soft.............................................................................. 817
Essential oils, water in (2.8.5.)................................................. 284 Extracts used in the preparation of herbal medicinal products
Ester value (2.5.2.) .................................................................... 161 for oral use, microbiological examination (2.6.31.) ........... 232
Estradiol benzoate .................................................................. 2404 Extracts used in the preparation of herbal medicinal products
Estradiol hemihydrate..................................................... 9.1-4151 for oral use, microbiological quality (5.1.8.) ....................... 591
Estradiol valerate .................................................................... 2407 Extracts, water for preparation of......................................... 3932
Estriol ................................................................................ 9.5-5651 Extraneous agents in viral vaccines for human use, tests for
Estrogens, conjugated............................................................. 2410 (2.6.16.) ........................................................................... 9.4-5111
Etacrynic acid.......................................................................... 2413 Extraneous agents : tests in batches of finished product of
Etamsylate................................................................................ 2414 avian live virus vaccines (2.6.25.) ......................................... 222
Etanercept ......................................................................... 9.5-5652 Extraneous agents : tests in seed lots of avian viral vaccines
Ethacridine lactate monohydrate.......................................... 2415 (2.6.24.) .................................................................................... 219
Ethambutol hydrochloride .................................................... 2416 Eye drops ................................................................................... 857
Ethanol (96 per cent).............................................................. 2417 Eye lotions ................................................................................. 857
Eye preparations ....................................................................... 857 Ferrous sulfate, dried.............................................................. 2466

Eye preparations, semi-solid ................................................... 858 Ferrous sulfate heptahydrate ................................................. 2467
Ferrum metallicum for homoeopathic preparations ......... 1601
F FET (18F) injection........................................................... 9.3-4797
F0 concept to steam sterilisation of aqueous preparations, Feverfew ................................................................................... 1355
application of (5.1.5.) ............................................................. 580 Fexofenadine hydrochloride.................................................. 2468
Factor II, human coagulation, assay of (2.7.18.)................... 266 Fibrinogen, human ................................................................. 2682
Factor IX, human coagulation .............................................. 2674 Fibrin sealant kit ..................................................................... 2469
Factor IX, human coagulation, assay of (2.7.11.) ................. 259 Filgrastim concentrated solution................................... 9.3-4899
Factor IX (rDNA) concentrated solution, human Films, orodispersible ....................................................... 9.3-4790
coagulation ..................................................................... 9.3-4915 Finasteride ............................................................................... 2473
Factor IX (rDNA) powder for solution for injection, human Fineness, powder (2.9.35.) ....................................................... 362
coagulation ..................................................................... 9.3-4920 Fipronil for veterinary use.............................................. 9.5-5661
Factor VIIa (rDNA) concentrated solution, human coagulation Fish oil, rich in omega-3 acids .............................................. 2474
......................................................................................... 9.5-5687 Flavoxate hydrochloride......................................................... 2476
Factor VII, human coagulation............................................. 2666 Flecainide acetate.................................................................... 2478
Factor VII, human coagulation, assay of (2.7.10.)................ 258 Fleeceflower root..................................................................... 1356
Factor VIII, human coagulation ........................................... 2672 Flocculation value (Lf) of diphtheria and tetanus toxins and
Factor VIII, human coagulation, assay of (2.7.4.) ................ 246 toxoids (Ramon assay) (2.7.27.)............................................ 273
Factor VIII (rDNA), human coagulation ............................ 2673 Flowability (2.9.16.).................................................................. 321
Factor X, human coagulation, assay of (2.7.19.) ................... 266 Flow cytometry 18
(2.7.24.).......................................................... 271
Factor XI, human coagulation .............................................. 2681 FLT ( F) injection .................................................................. 1127
Factor XI, human coagulation, assay of (2.7.22.) ................. 269 Flubendazole ........................................................................... 2479
Falling ball and automatic rolling ball viscometer methods Flucloxacillin magnesium octahydrate ................................ 2480
(2.2.49.) ........................................................................... 9.3-4690 Flucloxacillin sodium ............................................................. 2482
Famotidine............................................................................... 2449 Fluconazole.............................................................................. 2484
Fat, hard ................................................................................... 2639 Flucytosine............................................................................... 2485
Fat, hard with additives.......................................................... 2640 Fludarabine phosphate...........................................................
18
2487
Fatty acids, composition by gas chromatography (2.4.22.).. 141 Fludeoxyglucose ( F) injection ............................................ 1135
Fatty acids in oils rich in omega-3 acids, composition of Fludrocortisone acetate.......................................................... 2489
(2.4.29.) .................................................................................... 155 Flumazenil ...............................................................................
11
2490
Fatty oils, alkaline impurities in (2.4.19.) .............................. 138 Flumazenil (N-[ C]methyl) injection.................................. 1137
Fatty oils and resinified essential oils in essential oils Flumequine.............................................................................. 2492
(2.8.7.) ...................................................................................... 284 Flumetasone pivalate.............................................................. 2493
Fatty oils, foreign oils in, by thin-layer chromatography Flunarizine dihydrochloride.................................................. 2494
(2.4.21.) .................................................................................... 141 Flunitrazepam ......................................................................... 2495
Fatty oils, identification by thin-layer chromatography Flunixin meglumine for veterinary use ............................... 2496
(2.3.2.) ...................................................................................... 126 Fluocinolone acetonide.......................................................... 2497
Fatty oils, sterols in (2.4.23.).................................................... 144 Fluocortolone pivalate............................................................ 2500
Fatty oils, vegetable................................................................... 848 Fluorescein .............................................................................. 2501
Fc function of immunoglobulin, test for (2.7.9.)......... 9.3-4724 Fluorescein 18
sodium ................................................................ 2502
Febantel for veterinary use .................................................... 2450 Fluoride ( F) solution for radiolabelling............................. 1139
Feeding stuffs for veterinary use, medicated, premixes for.. 875 Fluorides (2.4.5.)....................................................................... 132
Felbinac .................................................................................... 2451 Fluorimetry (2.2.21.) 18
.................................................................. 36
Feline calicivirosis vaccine (inactivated).............................. 1053 Fluorocholine ( F) injection.................................................
18
1139
Feline calicivirosis vaccine (live)........................................... 1054 Fluorodeoxythymidine ( F) injection ................................. 1127
18
Feline chlamydiosis vaccine (inactivated) ........................... 1055 Fluorodopa ( F) (prepared by electrophilic substitution)
Feline infectious enteritis (feline panleucopenia) vaccine injection ................................................................................. 1141
18
(inactivated) .......................................................................... 1056 Fluoroethyl-L-tyrosine18 ( F) injection ........................... 9.3-4797
Feline infectious enteritis (feline panleucopenia) vaccine Fluoromisonidazole ( F) injection....................................... 1146
(live) ....................................................................................... 1057 Fluorouracil ...................................................................... 9.2-4545
Feline leukaemia vaccine (inactivated) ................................ 1058 Fluoxetine hydrochloride....................................................... 2505
Feline panleucopenia vaccine (inactivated)......................... 1056 Flupentixol dihydrochloride.................................................. 2507
Feline panleucopenia vaccine (live) ..................................... 1057 Fluphenazine decanoate......................................................... 2509
Feline viral rhinotracheitis vaccine (inactivated) ............... 1059 Fluphenazine dihydrochloride .............................................. 2510
Feline viral rhinotracheitis vaccine (live) ............................ 1060 Fluphenazine enantate ........................................................... 2512
Felodipine ................................................................................ 2452 Flurazepam monohydrochloride .......................................... 2513
Felypressin ............................................................................... 2454 Flurbiprofen............................................................................. 2514
Fenbendazole for veterinary use........................................... 2455 Fluspirilene .............................................................................. 2516
Fenbufen .................................................................................. 2456 Flutamide................................................................................. 2517
Fennel, bitter ........................................................................... 1352 Fluticasone propionate........................................................... 2518
Fennel, sweet ........................................................................... 1353 Flutrimazole ............................................................................ 2520
Fenofibrate............................................................................... 2457 Fluvastatin sodium ................................................................. 2521
Fenoterol hydrobromide ........................................................ 2458 Fluvoxamine 18
maleate.............................................................. 2523
Fentanyl............................................................................. 9.4-5373 FMISO ( F) injection ............................................................ 1146
Fentanyl citrate................................................................. 9.4-5374 Foams, cutaneous ..................................................................... 864
Fenticonazole nitrate .............................................................. 2462 Foams, intrauterine .................................................................. 862
Fenugreek ................................................................................ 1354 Foams, medicated ..................................................................... 859
Fermentation, products of ....................................................... 830 Foams, rectal ............................................................................. 882
Ferric chloride hexahydrate................................................... 2463 Foams, vaginal........................................................................... 889
Ferrous fumarate..................................................................... 2464 Folic acid hydrate............................................................. 9.5-5662
Ferrous gluconate ................................................................... 2465 Folinate, calcium..................................................................... 1914
Follitropin ......................................................................... 9.5-5663

Follitropin concentrated solution .................................. 9.5-5669 Gas-gangrene antitoxin (novyi) ............................................ 1116
Foot-and-mouth disease (ruminants) vaccine Gas-gangrene antitoxin (perfringens).................................. 1117
(inactivated) ................................................................... 9.5-5597 Gas-gangrene antitoxin (septicum)...................................... 1118
Foreign esters in essential oils (2.8.6.).................................... 284 Gas pycnometric density of solids (2.9.23.)........................... 339
Foreign matter (2.8.2.) ............................................................. 283 Gastro-resistant capsules ................................................ 9.4-5288
Foreign oils in fatty oils by thin-layer chromatography Gastro-resistant granules ......................................................... 860
(2.4.21.) .................................................................................... 141 Gastro-resistant tablets ................................................... 9.3-4791
Formaldehyde, free (2.4.18.).................................................... 137 Gefitinib ............................................................................ 9.3-4906
Formaldehyde solution (35 per cent) ................................... 2537 Gelatin............................................................................... 9.3-4907
Formic acid....................................................................... 9.4-5376 Gels............................................................................................. 884
Formoterol fumarate dihydrate............................................. 2537 Gels for injections..................................................................... 873
Foscarnet sodium hexahydrate ...................................... 9.2-4546 Gemcitabine hydrochloride................................................... 2570
Fosfomycin calcium................................................................ 2541 Gemfibrozil....................................................................... 9.5-5679
Fosfomycin sodium ................................................................ 2542 General notices (1.) ......................................................... 9.2-4275
Fosfomycin trometamol......................................................... 2543 General texts on biological products (5.2.) ........................... 599
Fosinopril sodium .................................................................. 2544 General texts on microbiology (5.1.) ..................................... 575
Fourstamen stephania root.................................................... 1357 Gene transfer medicinal products for human use (5.14.).... 739
Fowl cholera vaccine (inactivated) ....................................... 1063 Gentamicin sulfate.................................................................. 2573
Fowl-pox vaccine (live) .......................................................... 1064 Gentian root ............................................................................ 1365
Framycetin sulfate .................................................................. 2547 Gentian tincture...................................................................... 1366
Frangula bark .......................................................................... 1358 Gestodene ................................................................................ 2575
Frangula bark dry extract, standardised .............................. 1359 Ginger ...................................................................................... 1367
Frankincense, Indian.............................................................. 1392 Gingival solutions ............................................................ 9.3-4788
Fraxinus rhynchophylla bark ................................................ 1360 Ginkgo dry extract, refined and quantified ......................... 1368
Free formaldehyde (2.4.18.)..................................................... 137 Ginkgo leaf .............................................................................. 1370
Freezing point (2.2.18.) .............................................................. 34 Ginseng .................................................................................... 1372
Fresh bilberry fruit dry extract, refined and standardised.. 1361 Ginseng dry extract ................................................................ 1374
Friability of granules and spheroids (2.9.41.)........................ 375 Glass containers for pharmaceutical use (3.2.1.) .................. 423
Friability of uncoated tablets (2.9.7.) ..................................... 312 Glibenclamide ......................................................................... 2577
Fructose.................................................................................... 2548 Gliclazide ................................................................................. 2578
Fucus ........................................................................................ 1405 Glimepiride ............................................................................. 2580
Fulvestrant ............................................................................... 2549 Glipizide................................................................................... 2582
Fumitory ........................................................................... 9.2-4465 Glossary (dosage forms) ................................................. 9.2-4423
Functional groups and ions, identification reactions of Glucagon, human ................................................................... 2584
(2.3.1.) ...................................................................................... 123 Glucoheptonate, calcium ....................................................... 1917
Functionality-related characteristics of excipients Glucosamine hydrochloride ........................................... 9.5-5680
(5.15.) .............................................................................. 9.2-4411 Glucosamine sulfate potassium chloride ...................... 9.5-5681
Fungicidal, bactericidal or yeasticidal activity of antiseptic Glucosamine sulfate sodium chloride........................... 9.5-5682
medicinal products, determination of (5.1.11.) ......... 9.2-4348 Glucose.............................................................................. 9.1-4167
Furosemide ....................................................................... 9.4-5377 Glucose, liquid ........................................................................ 2590
Furunculosis vaccine (inactivated, oil-adjuvanted, injectable) Glucose, liquid, spray-dried .................................................. 2591
for salmonids.................................................................. 9.2-4427 Glucose monohydrate ..................................................... 9.1-4168
Fusidate, sodium..................................................................... 3579 Glutamic acid .......................................................................... 2593
Fusidic acid.............................................................................. 2552 Glutathione.............................................................................. 2594
Glycan analysis of glycoproteins (2.2.59.) ............................. 103
G Glycerol.................................................................................... 2595
Gabapentin ....................................................................... 9.4-5381 Glycerol (85 per cent) ............................................................ 2597
Gadobutrol monohydrate ............................................... 9.3-4905 Glycerol dibehenate................................................................ 2598
Gadodiamide hydrate...................................................... 9.1-4165 Glycerol distearate .................................................................. 2599
Galactose.................................................................................. 2562 Glycerol formal ....................................................................... 2600
Galantamine hydrobromide .................................................. 2563 Glycerol monocaprylate......................................................... 2601
Gallium (67Ga) citrate injection ............................................ 1148 Glycerol monocaprylocaprate ............................................... 2602
Gallium (68Ga) chloride solution for radiolabelling ........... 1148 Glycerol monolinoleate.......................................................... 2603
Gallium (68Ga) DOTATOC injection ................................... 1150 Glycerol mono-oleate ............................................................. 2604
Gallium (68Ga) edotreotide injection ................................... 1150 Glycerol monostearate 40-55 ......................................... 9.2-4551
Gammacyclodextrin ........................................................ 9.4-5382 Glycerol triacetate................................................................... 3825
Gammadex ....................................................................... 9.4-5382 Glyceryl trinitrate solution .................................................... 2606
Ganciclovir .............................................................................. 2566 Glycine ..................................................................................... 2608
Gargles .............................................................................. 9.3-4788 Glycoproteins, glycan analysis of (2.2.59.) ............................ 103
Garlic for homoeopathic preparations................................. 1590 Glycopyrronium bromide...................................................... 2609
Garlic powder.......................................................................... 1364 Glycyrrhizate ammonium...................................................... 1716
Gas adsorption, specific surface area by (2.9.26.)................. 344 Goldenrod................................................................................ 1375
Gas chromatography (2.2.28.)................................................... 44 Goldenrod, European............................................................. 1376
Gas detector tubes (2.1.6.) .............................................. 9.3-4685 Goldenseal rhizome................................................................ 1378
Gases, carbon dioxide in (2.5.24.) .......................................... 168 Gonadorelin acetate......................................................... 9.4-5383
Gases, carbon monoxide in (2.5.25.)...................................... 169 Gonadotrophin, chorionic.............................................. 9.4-5385
Gases, nitrogen monoxide and nitrogen dioxide in Gonadotrophin, equine serum, for veterinary use............. 2613
(2.5.26.) .................................................................................... 170 Goserelin.................................................................................. 2614
Gases, nitrous oxide in (2.5.35.) ............................................. 176 Grafted copolymer, macrogol poly(vinyl alcohol) ............. 2945
Gases, oxygen in (2.5.27.) ........................................................ 170 Gramicidin .............................................................................. 2616
Gases, water in (2.5.28.)........................................................... 170 Granisetron hydrochloride.................................................... 2617
Gas-gangrene antitoxin, mixed ............................................. 1116 Granules..................................................................................... 860
Granules and powders for oral solutions and Heavy metals (2.4.8.) ................................................................ 133
suspensions..................................................................... 9.3-4786 Heavy metals in herbal drugs and herbal drug preparations
Granules and powders for syrups .................................. 9.3-4787 (2.4.27.) .................................................................................... 152
Granules and spheroids, friability of (2.9.41.)....................... 375 Hedera helix for homoeopathic preparations ..................... 1602
Granules, coated ....................................................................... 860 Helium ..................................................................................... 2642
Granules, effervescent .............................................................. 860 Heparin, assay of (2.7.5.) ................................................ 9.1-4049
Granules, gastro-resistant ........................................................ 860 Heparin calcium .............................................................. 9.3-4911
Granules, modified-release...................................................... 860 Heparin in coagulation factors, assay of (2.7.12.)................. 259
Greater celandine.................................................................... 1380 Heparins, low-molecular-mass ............................................. 2646
Green tea........................................................................... 9.4-5304 Heparin sodium ............................................................... 9.3-4913
Griseofulvin............................................................................. 2619 Hepatitis A immunoglobulin, human.................................. 2683
Guaiacol ................................................................................... 2620 Hepatitis A (inactivated, adsorbed) and typhoid polysaccharide
Guaifenesin....................................................................... 9.4-5385 vaccine ..................................................................................... 929
Guanethidine monosulfate .................................................... 2623 Hepatitis A (inactivated) and hepatitis B (rDNA) vaccine
Guar.......................................................................................... 1381 (adsorbed) ............................................................................... 930
Guarana............................................................................. 9.4-5306 Hepatitis A vaccine, assay of (2.7.14.).................................... 262
Guar galactomannan .............................................................. 2623 Hepatitis A vaccine (inactivated, adsorbed).......................... 931
Guidelines for using the test for bacterial endotoxins Hepatitis A vaccine (inactivated, virosome) ......................... 932
(5.1.10.) .................................................................................... 593 Hepatitis B immunoglobulin for intravenous administration,
Guidelines for using the test for sterility (5.1.9.) .................. 592 human .................................................................................... 2684
Hepatitis B immunoglobulin, human .................................. 2684
H Hepatitis B (rDNA), diphtheria and tetanus vaccine
Haemagglutinins, anti-A and anti-B (2.6.20.)....................... 213 (adsorbed) ............................................................................... 901
Haematopoietic products, numeration of CD34/CD45+ cells Hepatitis B (rDNA), diphtheria, tetanus and pertussis
in (2.7.23.) ............................................................................... 269 (acellular, component) vaccine (adsorbed)......................... 910
Haematopoietic progenitor cells, human, colony-forming cell Hepatitis B (rDNA), diphtheria, tetanus, pertussis (acellular,
assay for (2.7.28.) .................................................................... 274 component), poliomyelitis (inactivated) and haemophilus
Haematopoietic stem cells, human....................................... 2682 type b conjugate vaccine (adsorbed)........................... 9.5-5585
Haemodiafiltration and haemofiltration, solutions for...... 2631 Hepatitis B vaccine (rDNA) .................................................... 935
Haemodialysis, concentrated solutions for ......................... 2628 Hepatitis B vaccine (rDNA), assay of (2.7.15.)...................... 263
Haemodialysis solutions, concentrated, water for Hepatitis C virus (HCV), validation of nucleic acid
diluting................................................................................... 2627 amplification techniques for the detection of HCV RNA in
Haemodialysis, solutions for ................................................. 2628 plasma pools : guidelines ....................................................... 214
Haemofiltration and haemodiafiltration, solutions for...... 2631 Hepatitis type I vaccine (live), viral, duck ........................... 1047
Haemophilus type b and meningococcal group C conjugate Heptaminol hydrochloride .................................................... 2649
vaccine ..................................................................................... 926 Herbal drug extracts................................................................. 815
Haemophilus type b (conjugate), diphtheria, tetanus Herbal drug extracts, monographs on (information chapter)
and pertussis (acellular, component) vaccine (5.23.) ....................................................................................... 807
(adsorbed) ...................................................................... 9.5-5583 Herbal drug preparations ........................................................ 819
Haemophilus type b (conjugate), diphtheria, tetanus, pertussis Herbal drugs..................................................................... 9.2-4419
(acellular, component) and poliomyelitis (inactivated) Herbal drugs and herbal drug preparations, heavy metals in
vaccine (adsorbed) ........................................................ 9.5-5587 (2.4.27.) .................................................................................... 152
Haemophilus type b (conjugate), diphtheria, tetanus, pertussis Herbal drugs and herbal drug preparations, high-performance
(acellular, component), hepatitis B (rDNA) and poliomyelitis thin-layer chromatography of (2.8.25.)................................ 295
(inactivated) vaccine (adsorbed) ................................. 9.5-5585 Herbal drugs, determination of aflatoxin B1 in (2.8.18.) ..... 289
Haemophilus type b (conjugate), diphtheria, tetanus, Herbal drugs, essential oils in (2.8.12.).................................. 285
pertussis (whole cell) and poliomyelitis (inactivated) vaccine Herbal drugs for homoeopathic preparations..................... 1570
(adsorbed) ...................................................................... 9.5-5590 Herbal drugs, microscopic examination of (2.8.23)............. 295
Haemophilus type b conjugate vaccine......................... 9.5-5592 Herbal drugs : sampling and sample preparation (2.8.20.).. 291
Haemorrhagic disease vaccine (inactivated), rabbit........... 1092 Herbal drugs, tannins in (2.8.14.)........................................... 288
Halofantrine hydrochloride................................................... 2633 Herbal drugs, test for aristolochic acids in (2.8.21) ............. 292
Haloperidol.............................................................................. 2634 Herbal medicinal products for oral use and extracts used in
Haloperidol decanoate ........................................................... 2636 their preparation, microbiological examination (2.6.31.).. 232
Halothane ................................................................................ 2637 Herbal medicinal products for oral use and extracts used in
Hamamelis bark...................................................................... 1382 their preparation, microbiological quality (5.1.8.) ............. 591
Hamamelis leaf........................................................................ 1383 Herbal preparations.................................................................. 819
Hard capsules ................................................................... 9.4-5287 Herbal substances ............................................................ 9.2-4419
Hard fat .................................................................................... 2639 Herbal teas................................................................................. 820
Hard fat with additives........................................................... 2640 Herbal teas, instant................................................................... 820
Hard paraffin........................................................................... 3268 Herpesvirus vaccine (inactivated), equine........................... 1050
Harmonisation, pharmacopoeial (5.8.)......................... 9.4-5265 Herpes zoster (shingles) vaccine (live)................................... 982
Hawthorn berries.................................................................... 1384 Hexamidine diisetionate ........................................................ 2650
Hawthorn leaf and flower ...................................................... 1385 Hexetidine ............................................................................... 2651
Hawthorn leaf and flower dry extract .................................. 1386 Hexosamines in polysaccharide vaccines (2.5.20.) ............... 167
Hawthorn leaf and flower liquid extract, quantified .......... 1387 Hexylresorcinol ....................................................................... 2652
HCP assays (2.6.34.) ........................................................ 9.1-4041 Highly purified water ............................................................. 3933
Healthy chicken flocks for the production of inactivated High-molecular-mass macrogols.......................................... 2952
vaccines for veterinary use (5.2.13.)..................................... 630 High-performance thin-layer chromatography of herbal drugs
Heavy bismuth subnitrate...................................................... 1853 and herbal drug preparations (2.8.25.) ................................ 295
Heavy kaolin............................................................................ 2847 Histamine (2.6.10.) ................................................................... 194
Heavy magnesium carbonate ................................................ 2955 Histamine dihydrochloride ................................................... 2653
Heavy magnesium oxide........................................................ 2962 Histamine for homoeopathic preparations .................. 9.1-4118
Histaminum for homoeopathic preparations .............. 9.1-4118

Histidine .................................................................................. 2654 Human coagulation factor IX (rDNA) powder for solution for
Histidine hydrochloride monohydrate ................................ 2655 injection .......................................................................... 9.3-4920
Homatropine hydrobromide ................................................. 2657 Human coagulation factor VII.............................................. 2666
Homatropine methylbromide ............................................... 2658 Human coagulation factor VIIa (rDNA) concentrated
Homoeopathic pillules, coated.............................................. 1587 solution ........................................................................... 9.5-5687
Homoeopathic pillules, impregnated ............................ 9.1-4118 Human coagulation factor VII, assay of (2.7.10.)................. 258
Homoeopathic preparations........................................... 9.1-4117 Human coagulation factor VIII ............................................ 2672
Homoeopathic preparations, acidum succinicum for.. 9.5-5621 Human coagulation factor VIII, assay of (2.7.4.) ................. 246
Homoeopathic preparations, Agaricus phalloides Human coagulation factor VIII (rDNA) ............................. 2673
for .................................................................................... 9.5-5621 Human coagulation factor X, assay of (2.7.19.) .................... 266
Homoeopathic preparations, Allium sativum for .............. 1590 Human coagulation factor XI ............................................... 2681
Homoeopathic preparations, ammonium carbonicum Human coagulation factor XI, assay of (2.7.22.) .................. 269
for .................................................................................... 9.3-4827 Human fibrinogen .................................................................. 2682
Homoeopathic preparations, Anacardium for.................... 1591 Human glucagon..................................................................... 2584
Homoeopathic preparations, Apis for.................................. 1592 Human haematopoietic progenitor cells, colony-forming cell
Homoeopathic preparations, arsenicum album for .... 9.5-5623 assay for (2.7.28.) .................................................................... 274
Homoeopathic preparations, aurum chloratum natronatum Human haematopoietic stem cells........................................ 2682
for ................................................................................... 9.5-5623 Human hepatitis A immunoglobulin................................... 2683
Homoeopathic preparations, barium chloratum for.......... 1594 Human hepatitis B immunoglobulin ................................... 2684
Homoeopathic preparations, Belladonna for............... 9.2-4492 Human hepatitis B immunoglobulin for intravenous
Homoeopathic preparations, cadmium sulfuricum for..... 1596 administration....................................................................... 2684
Homoeopathic preparations, calcium fluoratum for .. 9.5-5624 Human insulin ........................................................................ 2768
Homoeopathic preparations, calcium iodatum for ............ 1596 Human measles immunoglobulin ........................................ 2685
Homoeopathic preparations, Cocculus for ......................... 1597 Human normal immunoglobulin for intramuscular
Homoeopathic preparations, Crocus for ............................. 1599 administration....................................................................... 2685
Homoeopathic preparations, cuprum aceticum for........... 1599 Human normal immunoglobulin for intravenous
Homoeopathic preparations, cuprum metallicum for....... 1600 administration....................................................................... 2687
Homoeopathic preparations, ferrum metallicum for ........ 1601 Human normal immunoglobulin for subcutaneous
Homoeopathic preparations, hedera helix for .................... 1602 administration....................................................................... 2689
Homoeopathic preparations, herbal drugs for.................... 1570 Human papillomavirus vaccine (rDNA) ............................... 936
Homoeopathic preparations, hydrastis canadensis for ...... 1603 Human plasma for fractionation .......................................... 2691
Homoeopathic preparations, hyoscyamus for .................... 1604 Human plasma (pooled and treated for virus
Homoeopathic preparations, hypericum for....................... 1605 inactivation) ................................................................... 9.2-4555
Homoeopathic preparations, kalium bichromicum for..... 1608 Human plasmin inhibitor, assay of (2.7.25.) ......................... 272
Homoeopathic preparations, magnesium fluoratum for ... 1609 Human protein C, assay of (2.7.30.)....................................... 276
Homoeopathic preparations, magnesium phosphoricum Human protein S, assay of (2.7.31.)........................................ 277
for ........................................................................................... 1609 Human prothrombin complex .............................................. 2695
Homoeopathic preparations, mother tinctures for ............ 1571 Human rabies immunoglobulin ........................................... 2696
Homoeopathic preparations, Nux-vomica for ............. 9.3-4829 Human rubella immunoglobulin ......................................... 2698
Homoeopathic preparations, petroleum rectificatum for.. 1612 Human tetanus immunoglobulin ......................................... 2698
Homoeopathic preparations, pillules for ............................. 1586 Human varicella immunoglobulin ....................................... 2700
Homoeopathic preparations, selenium for................... 9.2-4494 Human varicella immunoglobulin for intravenous
Homoeopathic preparations, Staphysagria for.................... 1612 administration....................................................................... 2700
Homoeopathic preparations, sulfur for ............................... 1614 Human von Willebrand factor .............................................. 2701
Homoeopathic preparations, Urtica dioica for ................... 1614 Human von Willebrand factor, assay of (2.7.21.) ................. 268
Homoeopathic stocks (methods of preparation of) and Hyaluronate, sodium.............................................................. 3583
potentisation .................................................................. 9.2-4479 Hyaluronidase ........................................................................ 2702
Homoepathic preparations, Ignatia for......................... 9.3-4827 Hydralazine hydrochloride............................................. 9.2-4557
Honey ....................................................................................... 2659 Hydrastis canadensis for homoeopathic preparations ....... 1603
Honey bee for homoeopathic preparations......................... 1592 Hydrochloric acid, concentrated .......................................... 2704
Hop strobile............................................................................. 1388 Hydrochloric acid, dilute ....................................................... 2704
Horse-chestnut........................................................................ 1389 Hydrochlorothiazide .............................................................. 2705
Horse-chestnut dry extract, standardised............................ 1390 Hydrocodone hydrogen tartrate 2.5-hydrate ...................... 2706
Host-cell protein assays (2.6.34.) ................................... 9.1-4041 Hydrocortisone ....................................................................... 2708
Houttuynia herb............................................................... 9.4-5307 Hydrocortisone acetate .......................................................... 2711
Human α-1-proteinase inhibitor .......................................... 2694 Hydrocortisone hydrogen succinate..................................... 2713
Human α-1-proteinase inhibitor, assay of (2.7.32.).............. 278 Hydrogenated arachis oil ....................................................... 1752
Human albumin injection, iodinated (125I) ......................... 1152 Hydrogenated castor oil......................................................... 1966
Human albumin solution ...................................................... 2660 Hydrogenated cottonseed oil................................................. 2173
Human anti-D immunoglobulin .......................................... 2662 Hydrogenated soya-bean oil.................................................. 3630
Human anti-D immunoglobulin, assay of (2.7.13.) ............. 260 Hydrogenated vegetable oils, nickel in (2.4.31.) .......... 9.4-5103
Human anti-D immunoglobulin for intravenous Hydrogenated wool fat........................................................... 3943
administration....................................................................... 2663 Hydrogen peroxide solution (30 per cent) .......................... 2715
Human antithrombin III, assay of (2.7.17.)........................... 265 Hydrogen peroxide solution (3 per cent) ............................ 2715
Human antithrombin III concentrate .................................. 2664 Hydromorphone hydrochloride............................................ 2716
Human C1-esterase inhibitor................................................ 2665 Hydrophobic colloidal silica.................................................. 3551
Human C1-esterase inhibitor, assay of (2.7.34.) ................... 278 Hydrous wool fat .................................................................... 3944
Human coagulation factor II, assay of (2.7.18.).................... 266 Hydroxocobalamin acetate .................................................... 2717
Human coagulation factor IX ............................................... 2674 Hydroxocobalamin chloride.................................................. 2718
Human coagulation factor IX, assay of (2.7.11.) .................. 259 Hydroxocobalamin sulfate..................................................... 2719
Human coagulation factor IX (rDNA) concentrated Hydroxycarbamide ................................................................. 2720
solution ........................................................................... 9.3-4915 Hydroxychloroquine sulfate ........................................... 9.3-4922
Hydroxyethylcellulose ..................................................... 9.3-4924
Hydroxyethylmethylcellulose ................................................ 3037 Immunonephelometry for vaccine component assay

Hydroxyethyl salicylate .......................................................... 2723 (2.7.35.) .................................................................................... 279
Hydroxyethyl starches ............................................................ 3649 Immunosera and vaccines, phenol in (2.5.15.) ..................... 166
Hydroxyl value (2.5.3.)............................................................. 161 Immunosera and vaccines, veterinary, evaluation of efficacy
Hydroxypropylbetadex........................................................... 2725 of (5.2.7.).................................................................................. 612
Hydroxypropylcellulose ......................................................... 2727 Immunosera and vaccines, veterinary, evaluation of safety
Hydroxypropylcellulose, low-substituted ..................... 9.5-5691 (5.2.6.) ...................................................................................... 610
Hydroxypropylmethylcellulose ............................................. 2738 Immunosera for human use, animal ...................................... 821
Hydroxypropylmethylcellulose phthalate ............................ 2740 Immunosera for veterinary use............................................... 823
Hydroxypropyl starch............................................................. 3645 Immunosera for veterinary use, evaluation of the safety of
Hydroxypropyl starch, pregelatinised .................................. 3647 each batch (5.2.9.)................................................................... 625
Hydroxyzine hydrochloride............................................ 9.5-5693 Implants ..................................................................................... 873
Hymecromone......................................................................... 2731 Impurities, elemental, determination of (2.4.20.)........ 9.5-5539
Hymenoptera venoms for allergen products....................... 2732 Impurities in substances for pharmaceutical use, control of
Hyoscine .................................................................................. 2733 (5.10.) ....................................................................................... 723
Hyoscine butylbromide................................................... 9.5-5694 Inactivated vaccines for veterinary use, healthy chicken flocks
Hyoscine hydrobromide ........................................................ 2735 for the production of (5.2.13.) .............................................. 630
Hyoscyamine sulfate............................................................... 2737 Indapamide.............................................................................. 2755
Hyoscyamus for homoeopathic preparations ..................... 1604 Indian frankincense................................................................ 1392
Hypericum............................................................................... 1526 Indicators, approximate pH of solutions (2.2.4.) .................... 25
Hypericum for homoeopathic preparations........................ 1605 Indigo plant leaf ...................................................................... 1393
Hypromellose .......................................................................... 2738 Indinavir sulfate ...................................................................... 2757
Hypromellose phthalate ......................................................... 2740 Indium (111In) chloride solution ........................................... 1153
Indium (111In) oxine solution ................................................ 1154
I Indium (111In) pentetate injection ........................................ 1154
Ibuprofen ................................................................................. 2745 Indometacin ............................................................................ 2759
Iceland moss............................................................................ 1392 Inductively coupled plasma-atomic emission spectrometry
ICH (5.8.).......................................................................... 9.4-5265 (2.2.57.) .................................................................................... 100
Ichthammol ............................................................................. 2747 Inductively coupled plasma-mass spectrometry (2.2.58.) ... 101
Identification (2.3.) ................................................................... 123 Infectious bovine rhinotracheitis vaccine (inactivated)..... 1067
Identification and control of residual solvents (2.4.24.) ...... 146 Infectious bovine rhinotracheitis vaccine (live).................. 1068
Identification of fatty oils by thin-layer chromatography Infectious bronchitis vaccine (inactivated), avian .............. 1007
(2.3.2.) ...................................................................................... 126 Infectious bronchitis vaccine (live), avian ........................... 1008
Identification of phenothiazines by thin-layer chromatography Infectious bursal disease vaccine (inactivated), avian........ 1010
(2.3.3.) ...................................................................................... 127 Infectious bursal disease vaccine (live), avian..................... 1011
Identification reactions of ions and functional groups Infectious chicken anaemia vaccine (live) .................... 9.5-5598
(2.3.1.) ...................................................................................... 123 Infectious encephalomyelitis vaccine (live), avian.............. 1013
Idoxuridine .............................................................................. 2748 Infectious enteritis vaccine (inactivated), feline ................. 1056
Ifosfamide ................................................................................ 2749 Infectious enteritis vaccine (live), feline .............................. 1057
Ignatia for homoeopathic preparations ........................ 9.3-4827 Infectious laryngotracheitis vaccine (live), avian ............... 1014
Imatinib mesilate ............................................................. 9.2-4561 Infectious rhinotracheitis vaccine (live), turkey ................. 1107
Imipenem monohydrate ................................................. 9.4-5391 Influenza vaccine (inactivated), equine ............................... 1051
Imipramine hydrochloride .................................................... 2754 Influenza vaccine (inactivated), porcine.............................. 1087
Immunochemical methods (2.7.1.) ........................................ 239 Influenza vaccine (live, nasal) ................................................. 939
Immunoglobulin for human use, anti-T lymphocyte, Influenza vaccine (split virion, inactivated) .......................... 942
animal .................................................................................... 1741 Influenza vaccine (surface antigen, inactivated)................... 943
Immunoglobulin for intramuscular administration, human Influenza vaccine (surface antigen, inactivated, prepared in
normal.................................................................................... 2685 cell cultures) ............................................................................ 945
Immunoglobulin for intravenous administration, human Influenza vaccine (surface antigen, inactivated, virosome).. 947
anti-D ..................................................................................... 2663 Influenza vaccine (whole virion, inactivated) ....................... 949
Immunoglobulin for intravenous administration, human Influenza vaccine (whole virion, inactivated, prepared in cell
hepatitis B.............................................................................. 2684 cultures) ................................................................................... 950
Immunoglobulin for intravenous administration, human Infrared absorption spectrophotometry (2.2.24.)................... 39
normal.................................................................................... 2687 Infusions .................................................................................... 872
Immunoglobulin for intravenous administration, human Inhalation gas, krypton (81mKr)............................................. 1159
varicella.................................................................................. 2700 Inhalation powders.......................................................... 9.4-5292
Immunoglobulin for subcutaneous administration, human Inhalation, preparations for ........................................... 9.4-5288
normal.................................................................................... 2689 Inhalation, preparations for : aerodynamic assessment of fine
Immunoglobulin, human anti-D .......................................... 2662 particles (2.9.18.) .................................................................... 323
Immunoglobulin, human anti-D, assay of (2.7.13.) ............. 260 Injectable insulin preparations.............................................. 2776
Immunoglobulin, human hepatitis A .................................. 2683 Injections ................................................................................... 871
Immunoglobulin, human hepatitis B................................... 2684 Injections, gels for..................................................................... 873
Immunoglobulin, human measles ........................................ 2685 Injections or infusions, concentrates for ............................... 872
Immunoglobulin, human rabies ........................................... 2696 Injections or infusions, powders for....................................... 872
Immunoglobulin, human rubella ......................................... 2698 Inositol, myo- .......................................................................... 2761
Immunoglobulin, human tetanus......................................... 2698 Inserts, ophthalmic................................................................... 858
Immunoglobulin, human varicella....................................... 2700 Instant herbal teas .................................................................... 820
Immunoglobulin, test for anticomplementary activity of Insulin aspart........................................................................... 2762
(2.6.17.) ........................................................................... 9.3-4713 Insulin, bovine ................................................................. 9.3-4929
Immunoglobulin, test for Fc function of (2.7.9.)......... 9.3-4724 Insulin glargine ................................................................ 9.5-5699
Immunological veterinary medicinal products, substances of Insulin, human........................................................................ 2768
animal origin for the production of (5.2.5.)........................ 608 Insulin injection, biphasic ..................................................... 2770
Insulin injection, biphasic isophane..................................... 2770

Insulin injection, isophane .................................................... 2771 Isopropyl isostearate........................................................ 9.2-4563

Insulin injection, soluble ....................................................... 2771 Isopropyl myristate................................................................. 2820
Insulin lispro ........................................................................... 2771 Isopropyl palmitate................................................................. 2821
Insulin, porcine ................................................................ 9.3-4931 Isosorbide dinitrate, diluted ................................................. 2821
Insulin preparations, injectable............................................. 2776 Isosorbide mononitrate, diluted .......................................... 2823
Insulin zinc injectable suspension ........................................ 2778 Isotretinoin ....................................................................... 9.5-5705
Insulin zinc injectable suspension (amorphous) ................ 2779 Isoxsuprine hydrochloride..................................................... 2826
Insulin zinc injectable suspension (crystalline) .................. 2779 Ispaghula husk ........................................................................ 1400
Interferon alfa-2 concentrated solution ............................... 2780 Ispaghula seed ......................................................................... 1400
Interferon beta-1a concentrated solution ..................... 9.5-5701 Isradipine ................................................................................. 2828
Interferon gamma-1b concentrated solution ...................... 2785 Itraconazole ............................................................................. 2829
Interferons, assay of (5.6.)........................................................ 689 Ivermectin................................................................................ 2832
International System (SI) units (1.) ............................... 9.2-4275 Ivy leaf ...................................................................................... 1401
Intramammary preparations for veterinary use ................... 861
Intraruminal delivery systems ....................................... 9.3-4785 J
Intrauterine capsules ................................................................ 862 Javanese turmeric.................................................................... 1544
Intrauterine foams .................................................................... 862 Java tea ..................................................................................... 1402
Intrauterine preparations for veterinary use ......................... 862 Josamycin................................................................................. 2837
Intrauterine solutions, suspensions ........................................ 862 Josamycin propionate............................................................. 2839
Intrauterine sticks..................................................................... 862 Juniper...................................................................................... 1404
Intrauterine tablets ................................................................... 862 Juniper oil ................................................................................ 1404
Intrinsic dissolution (2.9.29.) .................................................. 347
In vitro method(s) for the quality control of vaccines,
K
substitution of in vivo method(s) by (5.2.14.)............ 9.3-4737
In vivo assay of poliomyelitis vaccine (inactivated) Kalium bichromicum for homoeopathic preparations ...... 1608
(2.7.20.) .................................................................................... 267 Kanamycin acid sulfate .......................................................... 2845
Iobenguane (123I) injection .................................................... 1155 Kanamycin monosulfate ........................................................ 2846
Iobenguane (131I) injection for diagnostic use..................... 1156 Kaolin, heavy........................................................................... 2847
Iobenguane (131I) injection for therapeutic use................... 1157 Kelp........................................................................................... 1405
Iobenguane sulfate for radiopharmaceutical preparations.. 9.2- Ketamine hydrochloride ........................................................ 2847
4435 Ketobemidone hydrochloride ............................................... 2848
Iodinated (125I) human albumin injection ........................... 1152 Ketoconazole ........................................................................... 2849
Iodinated povidone ................................................................ 3394 Ketoprofen ............................................................................... 2851
Iodine ....................................................................................... 2788 Ketorolac trometamol ............................................................ 2853
Iodine value (2.5.4.).................................................................. 161 Ketotifen hydrogen fumarate ................................................ 2854
Iodixanol.................................................................................. 2788 Knotgrass ................................................................................. 1406
Iodohippurate (sodium) dihydrate for radiopharmaceutical Krypton (81mKr) inhalation gas ............................................. 1159
preparations ................................................................... 9.2-4438 Kudzuvine root ................................................................ 9.3-4816
Iodomethylnorcholesterol (131I) injection............................ 1159 Kudzuvine root, Thomson.............................................. 9.3-4820
Iohexol...................................................................................... 2791
Ionic concentration, potentiometric determination of using L
ion-selective electrodes (2.2.36.)............................................. 60 Labetalol hydrochloride.................................................. 9.4-5397
Ions and functional groups, identification reactions of Lacosamide....................................................................... 9.5-5709
(2.3.1.) ...................................................................................... 123 Lactic acid................................................................................ 2860
Ion-selective electrodes, potentiometric determination of Lactic acid, (S)- ....................................................................... 2861
ionic concentration (2.2.36.) ................................................... 60 Lactitol monohydrate ...................................................... 9.3-4939
Iopamidol................................................................................. 2795 Lactobionic acid...................................................................... 2863
Iopanoic acid ........................................................................... 2797 Lactose .............................................................................. 9.3-4940
Iopromide ................................................................................ 2798 Lactose monohydrate ...................................................... 9.3-4942
Iotrolan .................................................................................... 2801 Lactulose ........................................................................... 9.5-5710
Ioxaglic acid............................................................................. 2803 Lactulose, liquid............................................................... 9.5-5712
Ipecacuanha liquid extract, standardised ............................ 1395 Lamivudine.............................................................................. 2871
Ipecacuanha, prepared ........................................................... 1395 Lamotrigine ............................................................................. 2873
Ipecacuanha root .................................................................... 1397 Lansoprazole .................................................................... 9.3-4945
Ipecacuanha tincture, standardised...................................... 1398 Largehead atractylodes rhizome ........................................... 1260
Ipratropium bromide ............................................................. 2805 Laryngotracheitis vaccine (live), infectious, avian ............. 1014
Irbesartan................................................................................. 2807 Laser light diffraction, particle size analysis by (2.9.31.) .... 349
Irinotecan hydrochloride trihydrate.............................. 9.3-4933 Laurilsulfate, sodium....................................................... 9.1-4200
Iron (2.4.9.)................................................................................ 136 Lauromacrogol 400................................................................. 2876
Iron for homoeopathic preparations.................................... 1601 Lauroyl macrogolglycerides............................................ 9.1-4173
Irrigation, preparations for...................................................... 880 Lavender flower ............................................................... 9.5-5614
Isatis root ................................................................................. 1399 Lavender oil...................................................................... 9.5-5615
Isoconazole .............................................................................. 2810 Lavender oil, spike........................................................... 9.5-5617
Isoconazole nitrate.................................................................. 2811 Lead in sugars (2.4.10.) ............................................................ 136
Isoelectric focusing (2.2.54.) ..................................................... 89 Leflunomide ............................................................................ 2879
Isoflurane ................................................................................. 2813 Lemon oil................................................................................. 1411
Isoleucine................................................................................. 2814 Lemon verbena leaf ................................................................ 1412
Isomalt .............................................................................. 9.4-5392 Leptospirosis vaccine (inactivated), bovine......................... 1019
Isoniazid............................................................................ 9.5-5703 Leptospirosis vaccine (inactivated), canine ......................... 1030
Isophane insulin injection ..................................................... 2771 Letrozole .................................................................................. 2880
Isoprenaline hydrochloride ................................................... 2817 Leucine..................................................................................... 2881
Isoprenaline sulfate................................................................. 2818 Leukaemia vaccine (inactivated), feline............................... 1058
Isopropyl alcohol .................................................................... 2819 Leuprorelin .............................................................................. 2883
Levamisole for veterinary use ............................................... 2885 Lubricant, silicone oil (3.1.8.).................................................. 408
Levamisole hydrochloride ..................................................... 2886 Lufenuron for veterinary use ................................................ 2929
Levetiracetam ................................................................... 9.4-5399 Lutetium (177Lu) solution for radiolabelling................. 9.3-4799
Levocabastine hydrochloride ......................................... 9.3-4946 Lycopus lucidus herb....................................................... 9.1-4105
Levocarnitine........................................................................... 2890 Lymecycline............................................................................. 2930
Levodopa ................................................................................. 2892 Lynestrenol .............................................................................. 2932
Levodropropizine.................................................................... 2893 Lyophilisates, oral ............................................................ 9.3-4792
Levofolinate pentahydrate, calcium...................................... 1926 Lysine acetate .......................................................................... 2933
Levomenthol............................................................................ 2894 Lysine hydrochloride.............................................................. 2935
Levomepromazine hydrochloride......................................... 2895
Levomepromazine maleate.................................................... 2896 M
Levomethadone hydrochloride ............................................. 2897 Macrogol 15 hydroxystearate ................................................ 2939
Levonorgestrel......................................................................... 2899 Macrogol 20 glycerol monostearate...................................... 2940
Levothyroxine sodium ........................................................... 2902 Macrogol 30 dipolyhydroxystearate ..................................... 2941
Levulinate dihydrate, calcium ............................................... 1928 Macrogol 40 sorbitol heptaoleate.......................................... 2941
Lidocaine ................................................................................. 2903 Macrogol 6 glycerol caprylocaprate...................................... 2939
Lidocaine hydrochloride........................................................ 2905 Macrogol cetostearyl ether .................................................... 2942
Light liquid paraffin ........................................................ 9.5-5737 Macrogolglycerol cocoates..................................................... 2948
Light magnesium carbonate .................................................. 2956 Macrogolglycerol hydroxystearate........................................ 2949
Light magnesium oxide.......................................................... 2963 Macrogolglycerol ricinoleate ................................................. 2950
Lime flower.............................................................................. 1414 Macrogol isotridecyl ether..................................................... 2943
Limit tests (2.4.) ........................................................................ 131 Macrogol lauryl ether............................................................. 2943
Limit tests, standard solutions for (4.1.2.).................... 9.4-5247 Macrogol oleate....................................................................... 2944
Lincomycin hydrochloride .................................................... 2906 Macrogol oleyl ether............................................................... 2945
Linen thread, sterile, in distributor for veterinary use ..... 1220 Macrogol poly(vinyl alcohol) grafted copolymer ............... 2945
Linoleoyl macrogolglycerides......................................... 9.1-4174 Macrogols ................................................................................ 2950
Linseed ..................................................................................... 1415 Macrogols, high-molecular-mass ......................................... 2952
Linseed oil, virgin ................................................................... 2908 Macrogol stearate.................................................................... 2947
Liothyronine sodium.............................................................. 2909 Macrogol stearyl ether ........................................................... 2947
Lipophilic solid dosage forms, dissolution test for Magaldrate............................................................................... 2953
(2.9.42.) .................................................................................... 377 Magnesium (2.4.6.)................................................................... 132
Liquid chromatography (2.2.29.) .............................................. 46 Magnesium acetate tetrahydrate ........................................... 2954
Liquid extraction preparations ............................................... 817 Magnesium aluminium silicate...................................... 9.3-4839
Liquid (fluid) extracts .............................................................. 817 Magnesium and alkaline-earth metals (2.4.7.)...................... 133
Liquid glucose ......................................................................... 2590 Magnesium aspartate dihydrate..................................... 9.3-4951
Liquid glucose, spray-dried ................................................... 2591 Magnesium carbonate, heavy................................................ 2955
Liquid lactulose................................................................ 9.5-5712 Magnesium carbonate, light .................................................. 2956
Liquid maltitol ........................................................................ 2974 Magnesium chloride 4.5-hydrate.......................................... 2957
Liquid paraffin ................................................................. 9.5-5737 Magnesium chloride hexahydrate ........................................ 2957
Liquid paraffin, light ....................................................... 9.5-5737 Magnesium citrate .................................................................. 2958
Liquid preparations for cutaneous application ..................... 864 Magnesium citrate dodecahydrate........................................ 2959
Liquid preparations for cutaneous application, Magnesium citrate nonahydrate ........................................... 2959
veterinary........................................................................ 9.3-4792 Magnesium fluoratum for homoeopathic preparations..... 1609
Liquid preparations for oral use .................................... 9.3-4785 Magnesium gluconate ............................................................ 2960
Liquids, clarity and degree of opalescence of (2.2.1.).. 9.2-4285 Magnesium glycerophosphate............................................... 2960
Liquid sorbitol (crystallising)................................................ 3626 Magnesium hydroxide............................................................ 2961
Liquid sorbitol (non-crystallising) ....................................... 3627 Magnesium lactate dihydrate ................................................ 2961
Liquid sorbitol, partially dehydrated.................................... 3628 Magnesium oxide, heavy ....................................................... 2962
Liquid sucrose .................................................................. 9.4-5448 Magnesium oxide, light.......................................................... 2963
Liquorice dry extract for flavouring purposes .................... 1415 Magnesium peroxide.............................................................. 2963
Liquorice root ......................................................................... 1416 Magnesium phosphoricum for homoeopathic
Lisinopril dihydrate ................................................................ 2910 preparations .......................................................................... 1609
Lithium carbonate .................................................................. 2912 Magnesium pidolate ............................................................... 2964
Lithium citrate ........................................................................ 2913 Magnesium stearate................................................................ 2965
L-Methionine ([11C]methyl) injection .................................. 1161 Magnesium sulfate heptahydrate .......................................... 2968
Lobeline hydrochloride.......................................................... 2913 Magnesium trisilicate ............................................................. 2968
Lomustine ................................................................................ 2914 Magnolia biondii flower bud.......................................... 9.3-4817
Long pepper ............................................................................ 1417 Magnolia officinalis bark ................................................ 9.2-4467
Loosestrife ............................................................................... 1419 Magnolia officinalis flower ............................................. 9.3-4819
Loperamide hydrochloride.................................................... 2915 Maize oil, refined .................................................................... 2969
Loperamide oxide monohydrate........................................... 2917 Maize starch ............................................................................ 2969
Lopinavir.................................................................................. 2918 Malathion................................................................................. 2970
Loratadine................................................................................ 2922 Maleic acid............................................................................... 2971
Lorazepam ............................................................................... 2924 Malic acid ................................................................................ 2972
Losartan potassium ................................................................ 2925 Mallow flower.......................................................................... 1424
Loss on drying (2.2.32.) .................................................. 9.4-5099 Mallow leaf .............................................................................. 1425
Loss on drying of extracts (2.8.17.) ........................................ 289 Maltitol..................................................................................... 2973
Lovage root.............................................................................. 1420 Maltitol, liquid ........................................................................ 2974
Lovastatin ................................................................................ 2927 Maltodextrin............................................................................ 2975
Low-molecular-mass heparins.............................................. 2646 Mandarin epicarp and mesocarp.......................................... 1426
Low-substituted hydroxypropylcellulose ...................... 9.5-5691 Mandarin oil............................................................................ 1428
Lozenges and pastilles ..................................................... 9.3-4789 Manganese gluconate ............................................................. 2976
Lozenges, compressed ..................................................... 9.3-4789

Manganese glycerophosphate, hydrated .............................. 2976 Meningococcal group C and haemophilus type b conjugate
Manganese sulfate monohydrate ................................... 9.2-4567 vaccine ..................................................................................... 926
Mannheimia vaccine (inactivated) for cattle....................... 1071 Meningococcal group C conjugate vaccine ........................... 956
Mannheimia vaccine (inactivated) for sheep ...................... 1072 Meningococcal polysaccharide vaccine ................................. 958
Mannitol ........................................................................... 9.2-4567 Menthol, racemic .................................................................... 2998
Maprotiline hydrochloride .................................................... 2980 Mepivacaine hydrochloride ................................................... 2999
Marbofloxacin for veterinary use ......................................... 2981 Meprobamate .......................................................................... 3001
Marek’s disease vaccine (live)................................................ 1073 Mepyramine maleate.............................................................. 3001
Marshmallow leaf ................................................................... 1429 Mercaptopurine ...................................................................... 3003
Marshmallow root .................................................................. 1430 Mercuric chloride ................................................................... 3003
Mass spectrometry (2.2.43.) ...................................................... 71 Mercury porosimetry, porosity and pore-size distribution of
Mass spectrometry, inductively coupled plasma- (2.2.58.).. 101 solids by (2.9.32.).................................................................... 352
Mass uniformity of delivered doses from multidose containers Meropenem trihydrate ........................................................... 3004
(2.9.27.) .................................................................................... 347 Mesalazine ............................................................................... 3005
Mass uniformity of single-dose preparations (2.9.5.) .......... 311 Mesna ................................................................................ 9.2-4569
Mastic ....................................................................................... 1430 Mesterolone ............................................................................. 3009
Mate leaf............................................................................ 9.4-5309 Mestranol ................................................................................. 3010
Materials based on non-plasticised poly(vinyl chloride) Metabisulfite, potassium ........................................................ 3386
for containers for non-injectable, aqueous solutions Metabisulfite, sodium............................................................. 3591
(3.1.10.) .................................................................................... 410 Metacresol................................................................................ 3011
Materials based on non-plasticised poly(vinyl chloride) for Metal catalyst or metal reagent residues (5.20.)........... 9.3-4759
containers for solid dosage forms for oral administration Metal catalyst or metal reagent residues, determination of
(3.1.11.) .................................................................................... 412 (2.4.20.) ........................................................................... 9.5-5539
Materials based on plasticised poly(vinyl chloride) for Metamizole sodium monohydrate........................................ 3012
containers for aqueous solutions for intravenous infusion Metered-dose preparations for inhalation, non-
(3.1.14.) .................................................................................... 416 pressurised...................................................................... 9.4-5291
Materials based on plasticised poly(vinyl chloride) for Metered-dose preparations for inhalation, pressurised ...... 9.4-
containers for human blood and blood components 5289
(3.1.1.1.) ................................................................................... 391 Metformin hydrochloride...................................................... 3014
Materials based on plasticised poly(vinyl chloride) for Methacrylate copolymer, basic butylated ..................... 9.4-5325
tubing used in sets for the transfusion of blood and blood Methacrylic acid - ethyl acrylate copolymer (1:1).............. 3015
components (3.1.1.2.)............................................................. 393 Methacrylic acid - ethyl acrylate copolymer (1:1) dispersion
Materials for containers for human blood and blood 30 per cent ............................................................................. 3017
components (3.1.1.)................................................................ 391 Methacrylic acid - methyl methacrylate copolymer (1:1).. 3018
Materials used for the manufacture of containers (3.1.)...... 391 Methacrylic acid - methyl methacrylate copolymer (1:2).. 3019
Matricaria flower .................................................................... 1431 Methadone hydrochloride ..................................................... 3020
Matricaria liquid extract ........................................................ 1433 Methane ................................................................................... 3021
Matricaria oil........................................................................... 1434 Methane intermix (2 per cent) in nitrogen .................. 9.3-4953
Meadowsweet .......................................................................... 1436 Methanesulfonate (methyl, ethyl and isopropyl) in active
Measles immunoglobulin, human ........................................ 2685 substances (2.5.38.)................................................................. 177
Measles, mumps and rubella vaccine (live)........................... 952 Methanesulfonic acid, methanesulfonyl chloride in
Measles, mumps, rubella and varicella vaccine (live) .......... 953 (2.5.39.) .................................................................................... 178
Measles vaccine (live)............................................................... 955 Methanesulfonic acid, methyl, ethyl and isopropyl
Measurement and detection of radioactivity (2.2.66.) ......... 113 methanesulfonate in (2.5.37.)................................................ 176
Measurement of consistency by penetrometry (2.9.9.)........ 313 Methanesulfonyl chloride in methanesulfonic acid
Mebendazole ........................................................................... 2983 (2.5.39.) .................................................................................... 178
Meclozine dihydrochloride............................................. 9.4-5403 Methanol.................................................................................. 3022
Medicated chewing gums ............................................... 9.3-4784 Methanol and 2-propanol, test for (2.9.11.) .......................... 318
Medicated chewing gums, dissolution test for (2.9.25.)....... 340 Methenamine .......................................................................... 3023
Medicated feeding stuffs for veterinary use, premixes for .. 875 Methionine .............................................................................. 3024
Medicated foams....................................................................... 859 Methionine ([11C]methyl) injection, L- ................................ 1161
Medicated plasters .................................................................... 884 Methionine, DL- ...................................................................... 3025
Medicated tampons .................................................................. 887 Methods in pharmacognosy (2.8.).......................................... 283
Medicated vaginal tampons..................................................... 889 Methods of preparation of homoeopathic stocks and
Medicinal air ........................................................................... 1653 potentisation .................................................................. 9.2-4479
Medicinal air, synthetic.......................................................... 1655 Methods of preparation of sterile products (5.1.1.) .... 9.2-4333
Medium-chain triglycerides .................................................. 3839 Methotrexate ........................................................................... 3026
Medronic acid for radiopharmaceutical preparations.. 9.2-4437 Methylcellulose ....................................................................... 3031
Medroxyprogesterone acetate................................................ 2985 Methyldopa.............................................................................. 3033
Mefenamic acid....................................................................... 2987 Methylene blue........................................................................ 3049
Mefloquine hydrochloride..................................................... 2989 Methylene chloride................................................................. 3034
Megestrol acetate .................................................................... 2990 Methylergometrine maleate .................................................. 3035
Meglumine............................................................................... 2992 Methyl, ethyl and isopropyl benzenesulfonate in active
Meldonium dihydrate............................................................. 2993 substances (2.5.41).................................................................. 180
Melilot ...................................................................................... 1437 Methyl, ethyl and isopropyl methanesulfonate in active
Melissa leaf .............................................................................. 1438 substances (2.5.38.)................................................................. 177
Melissa leaf dry extract .......................................................... 1439 Methyl, ethyl and isopropyl methanesulfonate in
Meloxicam ............................................................................... 2994 methanesulfonic acid (2.5.37.) .............................................. 176
Melphalan ................................................................................ 2996 Methyl, ethyl and isopropyl toluenesulfonate in active
Melting point - capillary method (2.2.14.) ................... 9.1-4037 substances (2.5.40.)................................................................. 179
Melting point - instantaneous method (2.2.16.) ..................... 33 Methylhydroxyethylcellulose................................................. 3037
Melting point - open capillary method (2.2.15.) .................... 32 Methyl methacrylate – methacrylic acid copolymer (1:1).. 3018
Menadione............................................................................... 2998 Methyl methacrylate - methacrylic acid copolymer (1:2).. 3019
Methyl nicotinate.................................................................... 3028 Molecular mass distribution in dextrans (2.2.39.).................. 62

Methyl parahydroxybenzoate................................................ 3029 Molgramostim concentrated solution........................... 9.5-5719
Methyl parahydroxybenzoate, sodium................................. 3591 Molsidomine ........................................................................... 3084
Methylpentoses in polysaccharide vaccines (2.5.21.) ........... 167 Molybdate dihydrate, sodium ............................................... 3592
Methylphenidate hydrochloride ........................................... 3037 Mometasone furoate........................................................ 9.5-5721
Methylphenobarbital .............................................................. 3039 Mometasone furoate monohydrate ............................... 9.5-5724
Methylprednisolone................................................................ 3040 Monoclonal antibodies for human use .................................. 826
Methylprednisolone acetate............................................ 9.4-5404 Monocyte-activation test (2.6.30.)................................. 9.2-4299
Methylprednisolone hydrogen succinate ............................. 3044 Monographs on herbal drug extracts (information chapter)
Methylpyrrolidone, N- ........................................................... 3046 (5.23.) ....................................................................................... 807
Methylrosanilinium chloride ......................................... 9.4-5406 Monophosphoryl lipid A, 3-O-desacyl-4- .......................... 2207
Methyl salicylate ..................................................................... 3030 Montelukast sodium........................................................ 9.1-4179
Methyltestosterone ................................................................. 3048 Morantel hydrogen tartrate for veterinary use ................... 3090
Methylthioninium chloride ................................................... 3049 Morphine hydrochloride ....................................................... 3091
Metixene hydrochloride......................................................... 3050 Morphine sulfate..................................................................... 3093
Metoclopramide............................................................... 9.4-5407 Moss, Iceland........................................................................... 1392
Metoclopramide hydrochloride monohydrate............. 9.4-5409 Mother tinctures for homoeopathic preparations .............. 1571
Metolazone .............................................................................. 3053 Motherwort ............................................................................. 1444
Metoprolol succinate .............................................................. 3055 Moulds for allergen products ................................................ 3094
Metoprolol tartrate ................................................................. 3056 Moutan bark..................................................................... 9.4-5311
Metrifonate .............................................................................. 3058 Mouthwashes.................................................................... 9.3-4788
Metronidazole ......................................................................... 3059 Moxidectin for veterinary use............................................... 3095
Metronidazole benzoate......................................................... 3060 Moxifloxacin hydrochloride .................................................. 3098
Mexiletine hydrochloride ...................................................... 3062 Moxonidine ............................................................................. 3100
Mianserin hydrochloride ....................................................... 3063 Mucoadhesive preparations............................................ 9.3-4789
Miconazole .............................................................................. 3064 Mullein flower ......................................................................... 1445
Miconazole nitrate.................................................................. 3066 Multidose containers, uniformity of mass of delivered doses
Microbial enumeration tests (microbiological examination of (2.9.27.) .................................................................................... 347
non-sterile products) (2.6.12.) .............................................. 195 Mumps, measles and rubella vaccine (live)........................... 952
Microbiological assay of antibiotics (2.7.2.) ................. 9.3-4719 Mumps, measles, rubella and varicella vaccine (live) .......... 953
Microbiological examination of cell-based preparations Mumps vaccine (live) ............................................................... 959
(2.6.27.) ........................................................................... 9.2-4297 Mupirocin ................................................................................ 3101
Microbiological examination of herbal medicinal products for Mupirocin calcium ................................................................. 3102
oral use and extracts used in their preparation (2.6.31.)... 232 Mycobacteria (2.6.2.)................................................................ 188
Microbiological examination of non-sterile products : Mycophenolate mofetil .......................................................... 3104
microbial enumeration tests (2.6.12.) .................................. 195 Mycophenolate sodium.......................................................... 3105
Microbiological examination of non-sterile products : test for Mycoplasma gallisepticum vaccine (inactivated) ............... 1075
specified micro-organisms (2.6.13.) ..................................... 199 Mycoplasmas (2.6.7.)................................................................ 188
Microbiological quality, alternative methods for control of myo-Inositol ............................................................................ 2761
(5.1.6.) ............................................................................. 9.2-4339 Myrrh ................................................................................ 9.2-4468
Microbiological quality of herbal medicinal products for oral Myrrh tincture ................................................................. 9.2-4469
use and extracts used in their preparation (5.1.8.)............. 591 Myxomatosis vaccine (live) for rabbits ................................ 1076
Microbiological quality of non-sterile pharmaceutical
preparations and substances for pharmaceutical use N
(5.1.4.) ...................................................................................... 579 Nabumetone ............................................................................ 3109
Microbiology, general texts on (5.1.) ..................................... 575 N-Acetyltryptophan ............................................................... 1639
Microcalorimetry and solution calorimetry, characterisation N-Acetyltyrosine ..................................................................... 1641
of crystalline solids by (2.2.61.) ............................................ 109 Nadolol..................................................................................... 3110
Microcrystalline cellulose ...................................................... 2010 Nadroparin calcium ........................................................ 9.5-5731
Microcrystalline cellulose and carmellose sodium ............ 3068 Naftidrofuryl hydrogen oxalate ............................................ 3113
Micro determination of water (2.5.32.)......................... 9.4-5107 Nalidixic acid .......................................................................... 3115
Microscopic examination of herbal drugs (2.8.23) .............. 295 Naloxone hydrochloride dihydrate................................ 9.3-4957
Microscopy, optical (2.9.37.) ................................................... 365 Naltrexone hydrochloride...................................................... 3118
Midazolam............................................................................... 3069 Names of herbal drugs used in traditional Chinese medicine
Milbemycin oxime for veterinary use ........................... 9.2-4570 (5.22.) .............................................................................. 9.4-5283
Milk thistle dry extract, refined and standardised ............. 1440 Nandrolone decanoate ........................................................... 3120
Milk thistle fruit...................................................................... 1441 Naphazoline hydrochloride ................................................... 3122
Minimising the risk of transmitting animal spongiform Naphazoline nitrate ................................................................ 3123
encephalopathy agents via human and veterinary medicinal Naproxen ................................................................................. 3124
products (5.2.8.)...................................................................... 613 Naproxen sodium ................................................................... 3126
Minocycline hydrochloride dihydrate........................... 9.2-4573 Narrow-leaved coneflower root ............................................ 1447
Minoxidil ................................................................................. 3072 Nasal drops and liquid nasal sprays ....................................... 867
Mint oil, partly dementholised ............................................. 1443 Nasal powders ........................................................................... 867
Mirtazapine ............................................................................. 3074 Nasal preparations .................................................................... 866
Misoprostol.............................................................................. 3075 Nasal preparations, semi-solid................................................ 868
Mites for allergen products ................................................... 3077 Nasal sprays (liquid) and nasal drops .................................... 866
Mitomycin ............................................................................... 3078 Nasal sticks ................................................................................ 868
Mitoxantrone hydrochloride ................................................. 3079 Nasal washes.............................................................................. 868
Modafinil ................................................................................. 3081 Nateglinide .............................................................................. 3128
Modified-release capsules ............................................... 9.4-5288 Near-infrared spectroscopy (2.2.40.)........................................ 64
Modified-release granules........................................................ 860 Nebulisation, characterisation of preparations for
Modified-release tablets .................................................. 9.3-4791 (2.9.44.) .................................................................................... 378
Mofetil mycophenolate .......................................................... 3104

Nebulisation, liquid preparations for ............................ 9.4-5289 Norgestrel ................................................................................ 3185

Neohesperidin-dihydrochalcone .......................................... 3130 Normal immunoglobulin for intramuscular administration,
Neomycin sulfate .................................................................... 3132 human .................................................................................... 2685
Neonatal piglet colibacillosis vaccine (inactivated)............ 1077 Normal immunoglobulin for intravenous administration,
Neonatal ruminant colibacillosis vaccine (inactivated)..... 1079 human .................................................................................... 2687
Neostigmine bromide............................................................. 3134 Normal immunoglobulin for subcutaneous administration,
Neostigmine metilsulfate ................................................ 9.5-5733 human .................................................................................... 2689
Neroli oil .................................................................................. 1449 Nortriptyline hydrochloride........................................... 9.3-4967
Netilmicin sulfate............................................................. 9.2-4579 Noscapine ................................................................................ 3187
Nettle leaf................................................................................. 1450 Noscapine hydrochloride hydrate......................................... 3188
Nettle root................................................................................ 1452 Notoginseng root .................................................................... 1454
Neurovirulence test of live viral vaccines (2.6.18.)............... 212 Nuclear magnetic resonance spectrometry (2.2.33.).............. 54
Nevirapine ........................................................................ 9.3-4959 Nuclear magnetic resonance spectrometry, peptide
Nevirapine hemihydrate ........................................................ 3139 identification by (2.2.64.)....................................................... 112
Newcastle disease vaccine (inactivated)............................... 1080 Nucleated cell count and viability (2.7.29.) ........................... 275
Newcastle disease vaccine (live)............................................ 1082 Nucleic acid amplification techniques (2.6.21.) .................... 214
Niaouli oil, cineole type ......................................................... 1453 Nucleic acids in polysaccharide vaccines (2.5.17.) ............... 166
Nicardipine hydrochloride ............................................. 9.3-4960 Numeration of CD34/CD45+ cells in haematopoietic products
Nicergoline ....................................................................... 9.4-5415 (2.7.23.) .................................................................................... 269
Nickel in hydrogenated vegetable oils (2.4.31.) ........... 9.4-5103 Nutmeg oil ............................................................................... 1455
Nickel in polyols (2.4.15.) ........................................................ 137 Nux-vomica for homoeopathic preparations ............... 9.3-4829
Niclosamide............................................................................. 3142 Nystatin.................................................................................... 3189
Niclosamide monohydrate .................................................... 3143
Nicorandil................................................................................ 3144 O
Nicotinamide........................................................................... 3145 O-Acetyl in polysaccharide vaccines (2.5.19.)....................... 167
Nicotine.................................................................................... 3146 Oak bark .................................................................................. 1456
Nicotine ditartrate dihydrate................................................. 3147 Octoxinol 10............................................................................ 3193
Nicotine resinate ..................................................................... 3148 Octyldodecanol ................................................................ 9.4-5421
Nicotinic acid ................................................................... 9.3-4961 Octyl gallate............................................................................. 3193
Nifedipine ................................................................................ 3151 Odour (2.3.4.)............................................................................ 127
Niflumic acid........................................................................... 3152 Odour and taste of essential oils (2.8.8.)................................ 284
Nifuroxazide............................................................................ 3154 Ofloxacin.................................................................................. 3195
Nikethamide............................................................................ 3155 Oils, essential............................................................................. 814
Nilutamide............................................................................... 3156 Oils, fatty, identification by thin-layer chromatography
Nimesulide............................................................................... 3157 (2.3.2.) ...................................................................................... 126
Nimodipine ............................................................................. 3158 Oils, fatty, vegetable.................................................................. 848
Nitrazepam .............................................................................. 3159 Oils rich in omega-3 acids, composition of fatty acids in
Nitrendipine ............................................................................ 3160 (2.4.29.) .................................................................................... 155
Nitric acid ................................................................................ 3161 Oils rich in omega-3 acids, total cholesterol in (2.4.32.) ..... 157
Nitric oxide.............................................................................. 3162 Ointments .................................................................................. 883
Nitrofural ................................................................................. 3163 Olanzapine............................................................................... 3196
Nitrofurantoin......................................................................... 3164 Oleic acid ................................................................................. 3197
Nitrogen ................................................................................... 3165 Oleoresins .................................................................................. 818
Nitrogen determination by sulfuric acid digestion (2.5.9.).. 164 Oleoyl macrogolglycerides ............................................. 9.1-4185
Nitrogen determination, primary aromatic amino (2.5.8.).. 163 Oleyl alcohol............................................................................ 3199
Nitrogen, low-oxygen............................................................. 3166 Olive leaf .................................................................................. 1456
Nitrogen monoxide and nitrogen dioxide in gases Olive leaf dry extract .............................................................. 1458
(2.5.26.) .................................................................................... 170 Olive oil, refined .............................................................. 9.3-4973
Nitroprusside, sodium ........................................................... 3593 Olive oil, virgin ................................................................ 9.3-4973
Nitrous oxide........................................................................... 3166 Olmesartan medoxomil ......................................................... 3201
Nitrous oxide in gases (2.5.35.)............................................... 176 Olsalazine sodium .................................................................. 3203
Nizatidine ................................................................................ 3167 Omega-3-acid ethyl esters 60 ................................................ 3205
N-Methylpyrrolidone ............................................................. 3046 Omega-3-acid ethyl esters 90 ......................................... 9.2-4583
NMR spectrometry (2.2.33.) ..................................................... 54 Omega-3 acids, composition of fatty acids in oils rich in
NMR spectrometry, peptide identification by (2.2.64.)....... 112 (2.4.29.) .................................................................................... 155
N,N-Dimethylaniline (2.4.26.) ................................................ 152 Omega-3 acids, fish oil rich in .............................................. 2474
Nomegestrol acetate ............................................................... 3169 Omega-3 acids, total cholesterol in oils rich in (2.4.32.) ..... 157
Nonoxinol 9............................................................................. 3170 Omega-3-acid triglycerides ................................................... 3209
Non-sterile pharmaceutical preparations and substances for Omeprazole ............................................................................. 3211
pharmaceutical use, microbiological quality of (5.1.4.)..... 579 Omeprazole magnesium ........................................................ 3213
Non-sterile products, microbiological examination of Omeprazole sodium ............................................................... 3214
(microbial enumeration tests) (2.6.12.) ............................... 195 Ondansetron hydrochloride dihydrate ................................ 3216
Non-sterile products, microbiological examination of (test for Opalescence of liquids, clarity and degree of (2.2.1.).. 9.2-4285
specified micro-organisms) (2.6.13.).................................... 199 Ophthalmic inserts ................................................................... 858
Noradrenaline hydrochloride......................................... 9.3-4962 Opium dry extract, standardised.......................................... 1459
Noradrenaline tartrate .................................................... 9.3-4964 Opium, prepared .................................................................... 1460
Norepinephrine hydrochloride ...................................... 9.3-4962 Opium, raw.............................................................................. 1461
Norepinephrine tartrate.................................................. 9.3-4964 Opium tincture, standardised ............................................... 1463
Norethisterone ........................................................................ 3174 Optical microscopy (2.9.37.) ................................................... 365
Norethisterone acetate ........................................................... 3175 Optical rotation (2.2.7.) .................................................. 9.5-5535
Norfloxacin....................................................................... 9.3-4965 Oral drops......................................................................... 9.3-4786
Norflurane ............................................................................... 3179 Oral lyophilisates ............................................................. 9.3-4792
Norgestimate ........................................................................... 3184
Oral powders............................................................................. 874 Paracetamol ...................................................................... 9.4-5429

Oral solutions, emulsions and suspensions.................. 9.3-4786 Paraffin, hard........................................................................... 3268
Oral use, liquid preparations for ................................... 9.3-4785 Paraffin, light liquid ........................................................ 9.5-5737
Oral use, veterinary semi-solid preparations for.................. 890 Paraffin, liquid ................................................................. 9.5-5737
Orbifloxacin for veterinary use............................................. 3217 Paraffin, white soft.................................................................. 3270
Orciprenaline sulfate .............................................................. 3219 Paraffin, yellow soft ................................................................ 3271
Oregano ................................................................................... 1464 Parahydroxybenzoate, butyl .................................................. 1892
Organ preservation, solutions for......................................... 3612 Parahydroxybenzoate, ethyl .................................................. 2428
Oriental cashew for homoeopathic preparations ............... 1591 Parahydroxybenzoate, methyl ............................................... 3029
Orientvine stem ...................................................................... 1466 Parahydroxybenzoate, propyl................................................ 3436
Orodispersible films ........................................................ 9.3-4790 Parahydroxybenzoate, sodium ethyl .................................... 3577
Orodispersible tablets ..................................................... 9.3-4792 Parahydroxybenzoate, sodium methyl................................. 3591
Oromucosal capsules....................................................... 9.3-4789 Parahydroxybenzoate, sodium propyl.................................. 3599
Oromucosal drops, oromucosal sprays and sublingual Parainfluenza virus vaccine (live), bovine ........................... 1021
sprays............................................................................... 9.3-4788 Parainfluenza virus vaccine (live), canine ........................... 1032
Oromucosal preparations ............................................... 9.3-4787 Paraldehyde ............................................................................. 3271
Oromucosal preparations, semi-solid ........................... 9.3-4788 Paramyxovirus 1 (Newcastle disease) vaccine (inactivated),
Oromucosal solutions and oromucosal suspensions... 9.3-4788 avian ....................................................................................... 1080
Oromucosal sprays, oromucosal drops and sublingual Paramyxovirus 1 (Newcastle disease) vaccine (live),
sprays............................................................................... 9.3-4787 avian ....................................................................................... 1082
Oromucosal suspensions and oromucosal solutions... 9.3-4787 Paramyxovirus 3 vaccine (inactivated) for turkeys, avian.. 1016
Orphenadrine citrate.............................................................. 3220 Parenteral preparations............................................................ 871
Orphenadrine hydrochloride ................................................ 3222 Parenteral preparations, test for extractable volume of
Oseltamivir phosphate .................................................... 9.2-4585 (2.9.17.) .................................................................................... 322
Osmolality (2.2.35.) .................................................................... 59 Parnaparin sodium ................................................................. 3272
Ouabain ................................................................................... 3225 Paroxetine hydrochloride ...................................................... 3272
Oxacillin sodium monohydrate ............................................ 3226 Paroxetine hydrochloride hemihydrate ............................... 3275
Oxaliplatin ............................................................................... 3228 Particles, fine, aerodynamic assessment of in preparations for
Oxazepam ................................................................................ 3231 inhalation (2.9.18.) ................................................................. 323
Oxcarbazepine......................................................................... 3232 Particle size analysis by laser light diffraction (2.9.31.)....... 349
Oxeladin hydrogen citrate ..................................................... 3234 Particle-size distribution estimation by analytical sieving
Oxfendazole for veterinary use ............................................. 3235 (2.9.38.) .................................................................................... 367
Oxidising substances (2.5.30.)................................................. 171 Particulate contamination : sub-visible particles (2.9.19.) ... 335
Oxitropium bromide .............................................................. 3236 Particulate contamination : visible particles (2.9.20.)........... 337
Oxolinic acid ........................................................................... 3237 Parvovirosis vaccine (inactivated), canine........................... 1033
Oxprenolol hydrochloride ..................................................... 3239 Parvovirosis vaccine (inactivated), porcine......................... 1089
Oxybuprocaine hydrochloride .............................................. 3239 Parvovirosis vaccine (live), canine........................................ 1034
Oxybutynin hydrochloride .................................................... 3241 Passion flower ......................................................................... 1469
Oxycodone hydrochloride ..................................................... 3242 Passion flower dry extract ..................................................... 1470
Oxygen ..................................................................................... 3243 Pastes .......................................................................................... 884
Oxygen (15O) ........................................................................... 1163 Pasteurella vaccine (inactivated) for sheep.......................... 1084
Oxygen (93 per cent).............................................................. 3244 Pastilles and lozenges ...................................................... 9.3-4789
Oxygen-flask method (2.5.10.)................................................ 164 Patches, cutaneous.................................................................... 882
Oxygen in gases (2.5.27.) ......................................................... 170 Patches, transdermal ................................................................ 873
Oxymetazoline hydrochloride............................................... 3245 Patches, transdermal, dissolution test for (2.9.4.)................. 309
Oxytetracycline dihydrate...................................................... 3247 Pea starch................................................................................. 3277
Oxytetracycline hydrochloride.............................................. 3249 Pefloxacin mesilate dihydrate................................................ 3277
Oxytocin .................................................................................. 3250 Pelargonium root .................................................................... 1470
Oxytocin concentrated solution............................................ 3251 Pemetrexed disodium heptahydrate..................................... 3279
Penbutolol sulfate ................................................................... 3281
P Penetrometry, measurement of consistency by (2.9.9.) ....... 313
Paclitaxel ........................................................................... 9.4-5425 Penicillamine........................................................................... 3282
Pale coneflower root............................................................... 1467 Penicillin G, benzathine......................................................... 1822
Palmitic acid............................................................................ 3259 Penicillin G potassium .................................................... 9.2-4505
Pamidronate disodium pentahydrate................................... 3259 Penicillin G, procaine...................................................... 9.2-4506
Pancreas powder ..................................................................... 3260 Penicillin G sodium......................................................... 9.2-4508
Pancuronium bromide ........................................................... 3263 Penicillin V ....................................................................... 9.1-4189
Panleucopenia vaccine (inactivated), feline ........................ 1056 Penicillin V potassium .................................................... 9.1-4190
Panleucopenia vaccine (live), feline ..................................... 1057 Pentaerythrityl tetranitrate, diluted...................................... 3284
Pansy, wild (flowering aerial parts)...................................... 1558 Pentamidine diisetionate ....................................................... 3286
Pantoprazole sodium sesquihydrate..................................... 3264 Pentazocine.............................................................................. 3287
Pantothenate, calcium ............................................................ 1929 Pentazocine hydrochloride .................................................... 3287
Papaverine hydrochloride...................................................... 3265 Pentazocine lactate ................................................................. 3288
Paper chromatography (2.2.26.) ............................................... 42 Pentetate sodium calcium for radiopharmaceutical
Papillomavirus vaccine (rDNA), human .............................. 936 preparations ................................................................... 9.3-4800
Paraben, butyl ......................................................................... 1892 Pentobarbital ........................................................................... 3289
Paraben, ethyl.......................................................................... 2428 Pentobarbital sodium ............................................................. 3289
Paraben, methyl ...................................................................... 3029 Pentoxifylline .......................................................................... 3290
Paraben, propyl ....................................................................... 3436 Pentoxyverine hydrogen citrate ............................................ 3292
Paraben, sodium ethyl............................................................ 3577 Peony root, red........................................................................ 1471
Paraben, sodium methyl ........................................................ 3591 Peony root, white .................................................................... 1472
Paraben, sodium propyl......................................................... 3599 Pepper ...................................................................................... 1474
Pepper, long............................................................................. 1417

Peppermint leaf................................................................ 9.2-4469 Phenylbutazone....................................................................... 3319

Peppermint leaf dry extract............................................ 9.2-4471 Phenylbutyrate, sodium ......................................................... 3595
Peppermint oil ........................................................................ 1478 Phenylephrine ......................................................................... 3321
Pepsin powder......................................................................... 3293 Phenylephrine hydrochloride................................................ 3322
Peptide identification by nuclear magnetic resonance Phenylmercuric acetate.......................................................... 3324
spectrometry (2.2.64.)............................................................ 112 Phenylmercuric borate........................................................... 3324
Peptide mapping (2.2.55.).......................................................... 90 Phenylmercuric nitrate .......................................................... 3325
Peptides, synthetic, acetic acid in (2.5.34.) ............................ 175 Phenylpropanolamine hydrochloride .................................. 3325
Perborate, hydrated sodium .................................................. 3594 Phenytoin................................................................................. 3326
Pergolide mesilate................................................................... 3295 Phenytoin sodium................................................................... 3327
Perindopril tert-butylamine .................................................. 3296 Phloroglucinol......................................................................... 3329
Peritoneal dialysis, solutions for ........................................... 3299 Phloroglucinol dihydrate ....................................................... 3330
Permethrin (25:75) ................................................................. 3301 pH of solutions, approximate (2.2.4.)....................................... 25
Peroxide value (2.5.5.).............................................................. 162 Pholcodine monohydrate ............................................... 9.1-4192
Perphenazine........................................................................... 3303 Phosphates (2.4.11.).................................................................. 136
Pertussis (acellular, component), diphtheria and tetanus Phospholipids for injection, egg .................................... 9.2-4533
vaccine (adsorbed) ................................................................. 902 Phospholipids for injection, soya................................... 9.4-5445
Pertussis (acellular, component), diphtheria and tetanus Phosphoric acid, concentrated.............................................. 3333
vaccine (adsorbed, reduced antigen(s) content)................. 903 Phosphoric acid, dilute .......................................................... 3334
Pertussis (acellular, component), diphtheria, tetanus and Phosphorus in polysaccharide vaccines (2.5.18.) ................. 166
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5583 pH, potentiometric determination of (2.2.3.) ......................... 24
Pertussis (acellular, component), diphtheria, tetanus and Phthalylsulfathiazole .............................................................. 3334
hepatitis B (rDNA) vaccine (adsorbed) ............................... 910 Physical and physicochemical methods (2.2.) ........................ 21
Pertussis (acellular, component), diphtheria, tetanus and Physostigmine salicylate ........................................................ 3335
poliomyelitis (inactivated) vaccine (adsorbed)................... 911 Phytomenadione ..................................................................... 3336
Pertussis (acellular, component), diphtheria, tetanus and Phytosterol............................................................................... 3337
poliomyelitis (inactivated) vaccine (adsorbed, reduced Picosulfate, sodium ................................................................ 3596
antigen(s) content) ................................................................. 913 Picotamide monohydrate....................................................... 3338
Pertussis (acellular, component), diphtheria, tetanus, Picric acid for homoeopathic preparations ......................... 1587
hepatitis B (rDNA), poliomyelitis (inactivated) and Piglet colibacillosis vaccine (inactivated), neonatal ........... 1077
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5585 Pillules for homoeopathic preparations............................... 1586
Pertussis (acellular, component), diphtheria, tetanus, Pillules, homoeopathic, coated ............................................. 1587
poliomyelitis (inactivated) and haemophilus type b conjugate Pillules, homoeopathic, impregnated............................ 9.1-4118
vaccine (adsorbed) ........................................................ 9.5-5587 Pilocarpine hydrochloride ..................................................... 3339
Pertussis toxin (residual) and pertussis toxoid (irreversibility Pilocarpine nitrate .................................................................. 3340
of) (2.6.33.).............................................................................. 235 Pimobendan for veterinary use...................................... 9.5-5738
Pertussis vaccine (acellular), assay of (2.7.16.)...................... 263 Pimozide .................................................................................. 3343
Pertussis vaccine (acellular, component, adsorbed)............. 960 Pindolol.................................................................................... 3344
Pertussis vaccine (acellular, co-purified, adsorbed) ............. 962 Pine (dwarf) oil....................................................................... 1340
Pertussis vaccine (whole cell, adsorbed)................................ 963 Pine sylvestris oil .................................................................... 1480
Pertussis vaccine (whole cell), assay of (2.7.7.) ..................... 252 Pioglitazone hydrochloride ................................................... 3345
Pertussis (whole cell), diphtheria and tetanus vaccine Pipemidic acid trihydrate ...................................................... 3346
(adsorbed) ............................................................................... 905 Piperacillin............................................................................... 3347
Pertussis (whole cell), diphtheria, tetanus and poliomyelitis Piperacillin sodium ................................................................ 3349
(inactivated) vaccine (adsorbed) .......................................... 920 Piperazine adipate .................................................................. 3351
Pertussis (whole cell), diphtheria, tetanus, poliomyelitis Piperazine citrate .................................................................... 3352
(inactivated) and haemophilus type b conjugate vaccine Piperazine hydrate .................................................................. 3352
(adsorbed) ...................................................................... 9.5-5590 Piracetam ................................................................................. 3353
Peru balsam ............................................................................. 1479 Pirenzepine dihydrochloride monohydrate ........................ 3354
Pessaries ..................................................................................... 888 Piretanide................................................................................. 3356
Pessaries and suppositories, disintegration of (2.9.2.) ......... 301 Pirfenidone .............................................................................. 3357
Pesticide residues (2.8.13.)....................................................... 286 Piroxicam................................................................................. 3358
Pethidine hydrochloride ........................................................ 3304 Pivampicillin ........................................................................... 3359
Petroleum rectificatum for homoeopathic preparations ... 1612 Pivmecillinam hydrochloride................................................ 3361
Pharmaceutical preparations.......................................... 9.5-5569 Plasma for fractionation, human.......................................... 2691
Pharmaceutical technical procedures (2.9.) .......................... 299 Plasma (pooled and treated for virus inactivation),
Pharmacognosy, methods in (2.8.)......................................... 283 human ............................................................................. 9.2-4555
Pharmacopoeial harmonisation (5.8.) .......................... 9.4-5265 Plasmid vectors for human use, bacterial cells used for the
Phenazone................................................................................ 3305 manufacture of........................................................................ 741
Pheniramine maleate.............................................................. 3306 Plasmin inhibitor, assay of human (2.7.25.).......................... 272
Phenobarbital .......................................................................... 3307 Plasters, medicated ................................................................... 882
Phenobarbital sodium............................................................ 3309 Plastic additives (3.1.13.) ......................................................... 414
Phenol ...................................................................................... 3310 Plastic containers and closures for pharmaceutical use
Phenol in immunosera and vaccines (2.5.15.) ...................... 166 (3.2.2.) ...................................................................................... 428
Phenolphthalein...................................................................... 3311 Plastic containers for aqueous solutions for infusion
Phenolsulfonphthalein ........................................................... 3311 (3.2.2.1.) ................................................................................... 429
Phenothiazines, identification by thin-layer chromatography Plastic containers for human blood and blood components,
(2.3.3.) ...................................................................................... 127 sterile (3.2.3.)........................................................................... 430
Phenoxyethanol ...................................................................... 3312 Plastic syringes, single-use, sterile (3.2.8.)............................. 434
Phenoxymethylpenicillin ................................................ 9.1-4189 Platycodon root................................................................ 9.4-5313
Phenoxymethylpenicillin potassium ............................. 9.1-4190 Pneumococcal polysaccharide conjugate vaccine
Phentolamine mesilate ........................................................... 3317 (adsorbed) ............................................................................... 965
Phenylalanine .......................................................................... 3318 Pneumococcal polysaccharide vaccine .................................. 967
Pneumonia vaccine (inactivated), porcine enzootic .......... 1086 Poly(vinyl alcohol)........................................................... 9.3-4979
Poliomyelitis (inactivated), diphtheria and tetanus vaccine Poly(vinyl alcohol) macrogol grafted copolymer ............... 2945
(adsorbed, reduced antigen(s) content)............................... 906 Poly(vinyl chloride) (non-plasticised) for containers for solid
Poliomyelitis (inactivated), diphtheria, tetanus and pertussis dosage forms for oral administration, materials based on
(acellular, component) vaccine (adsorbed)......................... 911 (3.1.11.) .................................................................................... 412
Poliomyelitis (inactivated), diphtheria, tetanus and pertussis Poly(vinyl chloride), non-plasticised, materials based on for
(acellular, component) vaccine (adsorbed, reduced antigen(s) containers for non-injectable aqueous solutions (3.1.10.).. 410
content).................................................................................... 913 Poly(vinyl chloride), plasticised, empty sterile containers of
Poliomyelitis (inactivated), diphtheria, tetanus and pertussis for human blood and blood components (3.2.4.) .............. 432
(whole cell) vaccine (adsorbed) ............................................ 920 Poly(vinyl chloride), plasticised, materials based on for
Poliomyelitis (inactivated), diphtheria, tetanus, pertussis containers for aqueous solutions for intravenous infusion
(acellular, component) and haemophilus type b conjugate (3.1.14.) .................................................................................... 416
vaccine (adsorbed) ........................................................ 9.5-5587 Poly(vinyl chloride), plasticised, materials based on for
Poliomyelitis (inactivated), diphtheria, tetanus, pertussis containers for human blood and blood components
(acellular, component), hepatitis B (rDNA) and haemophilus (3.1.1.1.) ................................................................................... 391
type b conjugate vaccine (adsorbed)........................... 9.5-5585 Poly(vinyl chloride), plasticised, materials based on for
Poliomyelitis (inactivated), diphtheria, tetanus, pertussis tubing used in sets for the transfusion of blood and blood
(whole cell) and haemophilus type b conjugate vaccine components (3.1.1.2.)............................................................. 393
(adsorbed) ...................................................................... 9.5-5590 Poly(vinyl chloride), plasticised, sterile containers of for
Poliomyelitis vaccine (inactivated) ......................................... 969 human blood containing anticoagulant solution (3.2.5.) .. 432
Poliomyelitis vaccine (inactivated), in vivo assay of Poppy petals, red..................................................................... 1492
(2.7.20.) .................................................................................... 267 Porcine actinobacillosis vaccine (inactivated) .................... 1085
Poliomyelitis vaccine (oral) ............................................ 9.1-4091 Porcine enzootic pneumonia vaccine (inactivated) ........... 1086
Pollens for allergen products................................................. 3362 Porcine influenza vaccine (inactivated) ............................... 1087
Poloxamers .............................................................................. 3363 Porcine insulin ................................................................. 9.3-4931
Polyacrylate dispersion 30 per cent...................................... 3365 Porcine parvovirosis vaccine (inactivated).......................... 1089
Polyamide 6/6 suture, sterile, in distributor for veterinary Porcine progressive atrophic rhinitis vaccine
use.................................................................................... 9.5-5609 (inactivated) .......................................................................... 1090
Polyamide 6 suture, sterile, in distributor for veterinary use Pore-size distribution of solids by mercury porosimetry,
......................................................................................... 9.5-5609 porosity and (2.9.32.) ............................................................. 352
Polyethyleneglycols................................................................. 2950 Poria ......................................................................................... 1484
Polyethyleneglycols, high-molecular-mass.......................... 2952 Porosimetry, mercury, porosity and pore-size distribution of
Polyethylene oxides, high-molecular-mass ......................... 2952 solids by (2.9.32.).................................................................... 352
Polyethylene terephthalate for containers for preparations not Porosity and pore-size distribution of solids by mercury
for parenteral use (3.1.15.) .................................................... 419 porosimetry (2.9.32.).............................................................. 352
Poly(ethylene terephthalate) suture, sterile, in distributor for Porosity of sintered-glass filters (2.1.2.)................................... 15
veterinary use................................................................. 9.5-5609 Porous solids including powders, wettability of (2.9.45.) .... 381
Poly(ethylene - vinyl acetate) for containers and tubing for Potassium (2.4.12.) ................................................................... 136
total parenteral nutrition preparations (3.1.7.).......... 9.2-4318 Potassium acetate.................................................................... 3375
Polyethylene with additives for containers for parenteral Potassium bromide................................................................. 3376
preparations and for ophthalmic preparations Potassium carbonate............................................................... 3377
(3.1.5.) ............................................................................. 9.4-5120 Potassium chloride ................................................................. 3377
Polyethylene without additives for containers for Potassium citrate..................................................................... 3378
parenteral preparations and for ophthalmic preparations Potassium clavulanate ............................................................ 3378
(3.1.4.) ............................................................................. 9.2-4310 Potassium clavulanate, diluted.............................................. 3381
Polygonum cuspidatum rhizome and root.......................... 1481 Potassium dichromate for homoeopathic preparations..... 1608
Polygonum orientale fruit .............................................. 9.2-4472 Potassium dihydrogen phosphate......................................... 3382
Polymorphism (5.9.)................................................................. 719 Potassium disulfite.................................................................. 3386
Polymyxin B sulfate ................................................................ 3366 Potassium hydrogen aspartate hemihydrate ....................... 3383
Polyolefins (3.1.3.) ........................................................... 9.4-5117 Potassium hydrogen carbonate ............................................. 3384
Polyoxyl castor oil................................................................... 2950 Potassium hydrogen tartrate ................................................. 3384
Polyoxyl hydrogenated castor oil .......................................... 2949 Potassium hydroxide ....................................................... 9.4-5430
Polyoxypropylene stearyl ether ...................................... 9.5-5739 Potassium iodide..................................................................... 3385
Polypropylene for containers and closures for parenteral Potassium metabisulfite ......................................................... 3386
preparations and ophthalmic preparations (3.1.6.)... 9.4-5123 Potassium nitrate .................................................................... 3386
Polysaccharide vaccines for human use, conjugated, carrier Potassium perchlorate............................................................ 3387
proteins for the production of (5.2.11.) ............................... 626 Potassium permanganate....................................................... 3388
Polysaccharide vaccines, hexosamines in (2.5.20.)............... 167 Potassium sodium tartrate tetrahydrate .............................. 3388
Polysaccharide vaccines, methylpentoses in (2.5.21.) .......... 167 Potassium sorbate ................................................................... 3389
Polysaccharide vaccines, nucleic acids in (2.5.17.) ............... 166 Potassium sulfate .................................................................... 3389
Polysaccharide vaccines, O-acetyl in (2.5.19.)....................... 167 Potato starch............................................................................ 3390
Polysaccharide vaccines, phosphorus in (2.5.18.)................. 166 Potentiometric determination of ionic concentration using
Polysaccharide vaccines, protein in (2.5.16.)......................... 166 ion-selective electrodes (2.2.36.)............................................. 60
Polysaccharide vaccines, ribose in (2.5.31.) .......................... 171 Potentiometric determination of pH (2.2.3.) .......................... 24
Polysaccharide vaccines, sialic acid in (2.5.23.) .................... 168 Potentiometric titration (2.2.20.).............................................. 35
Polysaccharide vaccines, uronic acids in (2.5.22.) ................ 168 Potentisation, methods of preparation of homoeopathic stocks
Polysorbate 20 ......................................................................... 3368 and................................................................................... 9.2-4479
Polysorbate 40 ......................................................................... 3369 Poultices..................................................................................... 884
Polysorbate 60 ......................................................................... 3370 Pour-on preparations ...................................................... 9.3-4792
Polysorbate 80 .................................................................. 9.2-4591 Povidone ........................................................................... 9.2-4592
Polystyrene sulfonate, sodium .............................................. 3597 Povidone, iodinated................................................................ 3394
Poly(vinyl acetate) ........................................................... 9.3-4977 Powdered cellulose ................................................................. 2013
Poly(vinyl acetate) dispersion 30 per cent .................... 9.3-4978 Powder fineness (2.9.35.) ......................................................... 362

Powder flow (2.9.36.)................................................................ 362 Propylene glycol dilaurate .............................................. 9.2-4598

Powders and granules for oral solutions and Propylene glycol monolaurate............................................... 3439
suspensions..................................................................... 9.3-4786 Propylene glycol monopalmitostearate................................ 3440
Powders and granules for syrups ................................... 9.3-4787 Propylene glycol monostearate ............................................. 3440
Powders and tablets for rectal solutions and suspensions... 882 Propyl gallate........................................................................... 3435
Powders, bulk density and tapped density of (2.9.34.)......... 359 Propyl parahydroxybenzoate................................................. 3436
Powders, ear .............................................................................. 856 Propyl parahydroxybenzoate, sodium ................................. 3599
Powders, effervescent ............................................................... 875 Propylthiouracil ...................................................................... 3441
Powders for cutaneous application......................................... 874 Propyphenazone ..................................................................... 3442
Powders for eye drops and powders for eye lotions ............. 858 Protamine sulfate.................................................................... 3443
Powders for injections or infusions........................................ 872 Protein assays, host-cell (2.6.34.) ................................... 9.1-4041
Powders for oral drops.................................................... 9.3-4786 Protein C, human, assay of (2.7.30.)....................................... 276
Powders, inhalation ......................................................... 9.4-5292 Protein in polysaccharide vaccines (2.5.16.) ......................... 166
Powders, nasal........................................................................... 867 Protein S, human, assay of (2.7.31.) ....................................... 277
Powders, oral ............................................................................. 874 Protein, total (2.5.33.) .............................................................. 172
Powders, wettability of porous solids including (2.9.45.) .... 381 Prothrombin complex, human.............................................. 2695
Pramipexole dihydrochloride monohydrate ....................... 3394 Protirelin.................................................................................. 3444
Pravastatin sodium ................................................................. 3396 Proxyphylline .......................................................................... 3445
Prazepam ................................................................................. 3397 Pseudoephedrine hydrochloride........................................... 3446
Praziquantel............................................................................. 3398 Psyllium seed........................................................................... 1486
Prazosin hydrochloride.......................................................... 3400 Pullulan.................................................................................... 3447
Prednicarbate .......................................................................... 3401 Purified water ................................................................... 9.4-5475
Prednisolone............................................................................ 3403 Purified water, highly............................................................. 3933
Prednisolone acetate............................................................... 3404 Purple coneflower herb.......................................................... 1486
Prednisolone pivalate ............................................................. 3406 Purple coneflower root .......................................................... 1488
Prednisolone sodium phosphate........................................... 3407 Pycnometric density of solids, gas (2.9.23.) .......................... 339
Prednisone........................................................................ 9.3-4980 Pygeum africanum bark ........................................................ 1490
Pregabalin ......................................................................... 9.2-4595 Pyrantel embonate.................................................................. 3448
Pregelatinised hydroxypropyl starch .................................... 3647 Pyrazinamide .......................................................................... 3449
Pregelatinised starch............................................................... 3649 Pyridostigmine bromide ........................................................ 3450
Prekallikrein activator (2.6.15.)............................................... 208 Pyridoxine hydrochloride...................................................... 3451
Premixes for medicated feeding stuffs for veterinary use ... 875 Pyrimethamine ....................................................................... 3453
Preparations for inhalation ............................................ 9.4-5288 Pyrogens (2.6.8.) ....................................................................... 193
Preparations for inhalation : aerodynamic assessment of fine Pyrophosphate (sodium) decahydrate for radiopharmaceutical
particles (2.9.18.) .................................................................... 323 preparations ................................................................... 9.2-4439
Preparations for irrigation....................................................... 880 Pyrrolidone.............................................................................. 3453
Preparations for nebulisation : characterisation (2.9.44.) .... 378
Pressurised pharmaceutical preparations.............................. 880 Q
Prilocaine.......................................................................... 9.4-5431 Quality control of vaccines, substitution of in vivo method(s)
Prilocaine hydrochloride ................................................ 9.4-5432 by in vitro method(s) for the (5.2.14.) ........................ 9.3-4737
Primaquine diphosphate........................................................ 3415 Quality of non-sterile pharmaceutical preparations and
Primary aromatic amino-nitrogen, determination of substances for pharmaceutical use, microbiological
(2.5.8.) ...................................................................................... 163 (5.1.4.) ...................................................................................... 579
Primary standards for volumetric solutions (4.2.1.) ... 9.4-5258 Quantified hawthorn leaf and flower liquid extract........... 1387
Primidone ......................................................................... 9.3-4982 Quetiapine fumarate .............................................................. 3457
Primula root ............................................................................ 1485 Quillaia bark............................................................................ 1491
Probenecid............................................................................... 3417 Quinapril hydrochloride........................................................ 3459
Procainamide hydrochloride................................................. 3418 Quinidine sulfate .................................................................... 3461
Procaine benzylpenicillin ............................................... 9.2-4506 Quinine hydrochloride .......................................................... 3463
Procaine hydrochloride.......................................................... 3419 Quinine sulfate........................................................................ 3464
Prochlorperazine maleate ...................................................... 3420
Products of fermentation......................................................... 830
R
Products of recombinant DNA technology........................... 836
Products with risk of transmitting agents of animal spongiform Rabbit haemorrhagic disease vaccine (inactivated) ........... 1092
encephalopathies .................................................................... 832 Rabeprazole sodium ............................................................... 3469
Progenitor cells, human haematopoietic, colony-forming cell Rabeprazole sodium hydrate................................................. 3470
assay for (2.7.28.) .................................................................... 274 Rabies immunoglobulin, human .......................................... 2696
Progesterone............................................................................ 3420 Rabies vaccine for human use prepared in cell cultures...... 976
Progressive atrophic rhinitis vaccine (inactivated), Rabies vaccine (inactivated) for veterinary use .................. 1093
porcine ................................................................................... 1090 Rabies vaccine (live, oral) for foxes and raccoon dogs ...... 1096
Proguanil hydrochloride................................................. 9.2-4597 Racecadotril............................................................................. 3472
Proline...................................................................................... 3424 Racementhol............................................................................ 2998
Promazine hydrochloride ...................................................... 3425 Racemic camphor ................................................................... 1934
Promethazine hydrochloride................................................. 3426 Racemic ephedrine hydrochloride ....................................... 2363
Propacetamol hydrochloride................................................. 3427 Racemic menthol .................................................................... 2998
Propafenone hydrochloride................................................... 3428 Racephedrine hydrochloride................................................. 2363
Propanol................................................................................... 3430 Raclopride ([11C]methoxy) injection.................................... 1165
Propanol and methanol, 2-, test for (2.9.11.) ........................ 318 Radioactivity, detection and measurement of (2.2.66.) ....... 113
Propantheline bromide .......................................................... 3431 Radionuclides, table of physical characteristics (5.7.).. 9.2-4385
Propofol ................................................................................... 3432 Radiopharmaceutical preparations ........................................ 832
Propranolol hydrochloride .................................................... 3434 Radiopharmaceutical preparations, chemical precursors
Propylene glycol...................................................................... 3437 for ............................................................................................. 813
Propylene glycol dicaprylocaprate........................................ 3438
Radiopharmaceutical preparations, copper tetramibi Ribwort plantain ..................................................................... 1497

tetrafluoroborate for...................................................... 9.2-4435 Rice starch ............................................................................... 3487
Radiopharmaceutical preparations, iobenguane sulfate Rifabutin .................................................................................. 3488
for .................................................................................... 9.2-4435 Rifampicin ............................................................................... 3489
Radiopharmaceutical preparations, medronic acid Rifamycin sodium................................................................... 3490
for .................................................................................... 9.2-4437 Rifaximin ................................................................................. 3492
Radiopharmaceutical preparations, pentetate sodium calcium Rilmenidine dihydrogen phosphate ..................................... 3494
for .................................................................................... 9.3-4800 Risedronate sodium 2.5-hydrate........................................... 3495
Radiopharmaceutical preparations, sodium iodohippurate Risperidone ............................................................................. 3496
dihydrate for................................................................... 9.2-4438 Ritonavir .................................................................................. 3498
Radiopharmaceutical preparations, sodium pyrophosphate Rivastigmine............................................................................ 3501
decahydrate for .............................................................. 9.2-4439 Rivastigmine hydrogen tartrate ............................................ 3503
Radiopharmaceutical preparations, tetra-O-acetyl-mannose Rizatriptan benzoate .............................................................. 3504
triflate for ....................................................................... 9.2-4443 Rocuronium bromide............................................................. 3506
Radiopharmaceuticals, extemporaneous preparation of Rolling ball (automatic) and falling ball viscometer methods
(5.19.) ....................................................................................... 767 (2.2.49.) ........................................................................... 9.3-4690
Raloxifene hydrochloride ...................................................... 3473 Roman chamomile flower...................................................... 1313
Raltegravir chewable tablets ........................................... 9.5-5743 Ropinirole hydrochloride ...................................................... 3508
Raltegravir potassium ..................................................... 9.4-5437 Ropivacaine hydrochloride monohydrate ........................... 3509
Raltegravir tablets ............................................................ 9.5-5744 Roselle ...................................................................................... 1498
Raman spectroscopy (2.2.48.) ................................................... 86 Rosemary leaf.......................................................................... 1499
Ramipril ................................................................................... 3475 Rosemary oil ........................................................................... 1500
Ramon assay, flocculation value (Lf) of diphtheria and tetanus Rosuvastatin calcium.............................................................. 3511
toxins and toxoids (2.7.27.) ................................................... 273 Rotating viscometer method - viscosity (2.2.10.) ................... 28
Ranitidine hydrochloride................................................ 9.2-4603 Rotation, optical (2.2.7.) ................................................. 9.5-5535
Rapeseed oil, refined .............................................................. 3479 Rotavirus diarrhoea vaccine (inactivated), calf .................. 1026
Raw materials of biological origin for the production of Rotavirus vaccine (live, oral)................................................... 978
cell-based and gene therapy medicinal products (5.2.12.).. 627 Round amomum fruit ............................................................ 1502
Reagents (4.1.1.)............................................................... 9.4-5131 Roxithromycin ........................................................................ 3513
Reagents (4.1.1.)............................................................... 9.5-5549 RRR-α-Tocopherol.................................................................. 3801
Reagents (4.)..................................................................... 9.4-5131 RRR-α-Tocopheryl acetate..................................................... 3804
Reagents, standard solutions, buffer solutions (4.1.) .. 9.5-5549 RRR-α-Tocopheryl hydrogen succinate ............................... 3808
Recombinant DNA technology, products of......................... 836 Rubber closures for containers for aqueous parenteral
Recommendations on dissolution testing (5.17.1.) .............. 761 preparations, for powders and for freeze-dried powders
Recommendations on methods for dosage forms testing (3.2.9.) ............................................................................. 9.5-5545
(5.17.) ....................................................................................... 761 Rubella immunoglobulin, human ........................................ 2698
Rectal capsules .......................................................................... 882 Rubella, measles and mumps vaccine (live) .......................... 952
Rectal foams .............................................................................. 882 Rubella, measles, mumps and varicella vaccine (live).......... 953
Rectal preparations................................................................... 881 Rubella vaccine (live) ............................................................... 981
Rectal preparations, semi-solid............................................... 882 Ruminant colibacillosis vaccine (inactivated), neonatal.... 1079
Rectal solutions and suspensions, powders and tablets for.. 881 Rupatadine fumarate ....................................................... 9.3-4987
Rectal solutions, emulsions and suspensions........................ 882 Rutoside trihydrate ................................................................. 3516
Rectal tampons.......................................................................... 882
Red peony root........................................................................ 1471 S
Red poppy petals..................................................................... 1492 Saccharin.................................................................................. 3521
Reference standards (5.12.) ............................................ 9.5-5563 Saccharin sodium ................................................................... 3522
Refractive index (2.2.6.) ............................................................. 26 Safety, viral (5.1.7.) ................................................................... 591
Relative density (2.2.5.).............................................................. 25 Safflower flower ...................................................................... 1503
Remifentanil hydrochloride ........................................... 9.2-4604 Safflower oil, refined............................................................... 3523
Repaglinide.............................................................................. 3479 Saffron for homoeopathic preparations............................... 1599
Reserpine ................................................................................. 3481 Sage leaf (Salvia officinalis) ................................................... 1505
Residual pertussis toxin and irreversibility of pertussis toxoid Sage leaf, three-lobed ............................................................. 1506
(2.6.33.) .................................................................................... 235 Sage oil, Spanish...................................................................... 1524
Residual solvents (5.4.) ................................................... 9.5-5553 Sage tincture ............................................................................ 1507
Residual solvents, identification and control (2.4.24.) ......... 146 Salbutamol ............................................................................... 3523
Residue on evaporation of essential oils (2.8.9.)................... 284 Salbutamol sulfate................................................................... 3526
Resistance to crushing of tablets (2.9.8.) ............................... 313 Salicylic acid ............................................................................ 3528
Resorcinol ................................................................................ 3481 Salmeterol xinafoate ............................................................... 3529
Respiratory syncytial virus vaccine (live), bovine .............. 1022 Salmonella Enteritidis vaccine (inactivated) for chickens.. 1097
Restharrow root ...................................................................... 1493 Salmonella Enteritidis vaccine (live, oral) for chickens ..... 1098
Retroviridae-derived vectors for human use......................... 746 Salmonella Typhimurium vaccine (inactivated) for
Rhatany root............................................................................ 1494 chickens ................................................................................. 1100
Rhatany tincture ..................................................................... 1495 Salmonella Typhimurium vaccine (live, oral) for
Rhinotracheitis vaccine (inactivated), bovine, infectious.. 1067 chickens ................................................................................. 1101
Rhinotracheitis vaccine (inactivated), viral, feline ............. 1059 Salmon oil, farmed ................................................................. 3531
Rhinotracheitis vaccine (live), bovine, infectious............... 1068 Salvia miltiorrhiza root and rhizome............................ 9.1-4108
Rhinotracheitis vaccine (live), infectious, turkey ............... 1107 Sanguisorba root..................................................................... 1509
Rhinotracheitis vaccine (live), viral, feline .......................... 1060 Saponification value (2.5.6.) .................................................... 163
Rhubarb ................................................................................... 1496 Saquinavir mesilate................................................................. 3533
Ribavirin .................................................................................. 3482 Saw palmetto extract .............................................................. 1509
Riboflavin................................................................................. 3484 Saw palmetto fruit .................................................................. 1512
Riboflavin sodium phosphate ............................................... 3485 Schisandra fruit................................................................ 9.1-4110
Ribose in polysaccharide vaccines (2.5.31.) .......................... 171

Scopolamine ............................................................................ 2733 Sodium aurothiomalate ......................................................... 3565

Scopolamine butylbromide ............................................ 9.5-5694 Sodium benzoate .................................................................... 3566
Scopolamine hydrobromide .................................................. 2735 Sodium bromide ..................................................................... 3567
Selamectin for veterinary use................................................ 3535 Sodium calcium edetate......................................................... 3568
Selegiline hydrochloride ........................................................ 3537 Sodium calcium pentetate for radiopharmaceutical
Selenium disulfide .................................................................. 3538 preparations ................................................................... 9.3-4800
Selenium for homoeopathic preparations .................... 9.2-4494 Sodium caprylate .................................................................... 3569
Selfheal fruit-spike, common ................................................ 1327 Sodium carbonate................................................................... 3570
Semi-micro determination of water (2.5.12.)............... 9.4-5107 Sodium carbonate decahydrate............................................. 3570
Semi-solid ear preparations..................................................... 856 Sodium carbonate monohydrate .......................................... 3571
Semi-solid eye preparations .................................................... 858 Sodium carboxymethylcellulose ........................................... 1958
Semi-solid intrauterine preparations ..................................... 862 Sodium carboxymethylcellulose, cross-linked.................... 2174
Semi-solid nasal preparations ................................................. 868 Sodium carboxymethylcellulose, low-substituted .............. 1958
Semi-solid oromucosal preparations............................. 9.3-4788 Sodium cetostearyl sulfate.............................................. 9.4-5443
Semi-solid preparations for cutaneous application .............. 882 Sodium chloride...................................................................... 3573
Semi-solid preparations for oral use, veterinary .................. 890 Sodium chromate (51Cr) sterile solution.............................. 1167
Semi-solid rectal preparations ................................................ 882 Sodium citrate ......................................................................... 3574
Semi-solid vaginal preparations ............................................. 889 Sodium cromoglicate ............................................................. 3574
Senega root .............................................................................. 1515 Sodium cyclamate................................................................... 3576
Senna leaf................................................................................. 1516 Sodium dihydrogen phosphate dihydrate ........................... 3577
Senna leaf dry extract, standardised .................................... 1517 Sodium disulfite ...................................................................... 3591
Senna pods, Alexandrian....................................................... 1518 Sodium ethyl parahydroxybenzoate..................................... 3577
Senna pods, Tinnevelly .......................................................... 1519 Sodium fluoride ...................................................................... 3579
Separation techniques, chromatographic (2.2.46.) ...... 9.2-4286 Sodium fluoride (18F) injection ............................................. 1168
Serine........................................................................................ 3539 Sodium fusidate ...................................................................... 3579
Sertaconazole nitrate .............................................................. 3540 Sodium glycerophosphate, hydrated .................................... 3582
Sertraline hydrochloride................................................. 9.4-5441 Sodium hyaluronate ............................................................... 3583
Sesame oil, refined........................................................... 9.1-4197 Sodium hydrogen carbonate ................................................. 3585
Sets for the transfusion of blood and blood components Sodium hydroxide .................................................................. 3585
(3.2.6.) ...................................................................................... 433 Sodium iodide......................................................................... 3586
Sevoflurane ....................................................................... 9.5-5749 Sodium iodide (123I) injection ............................................... 1169
Shampoos .................................................................................. 864 Sodium iodide (123I) solution for radiolabelling ................. 1170
Shellac ..................................................................................... 3547 Sodium iodide (131I) capsules for diagnostic use ................ 1170
Shingles (herpes zoster) vaccine (live) ................................... 982 Sodium iodide (131I) capsules for therapeutic use .............. 1171
Sialic acid in polysaccharide vaccines (2.5.23.)..................... 168 Sodium iodide (131I) solution ................................................ 1172
Siam benzoin........................................................................... 1272 Sodium iodide (131I) solution for radiolabelling ................. 1173
Siam benzoin tincture ............................................................ 1274 Sodium iodohippurate (123I) injection ................................. 1175
Sieves (2.1.4.)............................................................................... 16 Sodium iodohippurate (131I) injection ................................. 1176
Sieve test (2.9.12.) ..................................................................... 319 Sodium iodohippurate dihydrate for radiopharmaceutical
Sieving, analytical, particle-size distribution estimation by preparations ................................................................... 9.2-4438
(2.9.38.) .................................................................................... 367 Sodium lactate solution ......................................................... 3586
SI (International System) units (1.) ............................... 9.2-4275 Sodium laurilsulfate ........................................................ 9.1-4200
Sildenafil citrate ............................................................... 9.2-4611 Sodium lauroylsarcosinate for external use......................... 3589
Silica, colloidal anhydrous..................................................... 3549 Sodium metabisulfite ............................................................. 3591
Silica, colloidal hydrated........................................................ 3550 Sodium methyl parahydroxybenzoate ................................. 3591
Silica, dental type.................................................................... 3550 Sodium molybdate (99Mo) solution (fission) ...................... 1176
Silica, hydrophobic colloidal ................................................. 3551 Sodium molybdate dihydrate................................................ 3592
Silicate, aluminium magnesium..................................... 9.3-4839 Sodium nitrite ......................................................................... 3593
Silicate, aluminium sodium................................................... 1686 Sodium nitroprusside............................................................. 3593
Silicone elastomer for closures and tubing (3.1.9.)............... 409 Sodium perborate, hydrated.................................................. 3594
Silicone oil used as a lubricant (3.1.8.)................................... 408 Sodium pertechnetate (99mTc) injection (accelerator-
Silk suture, sterile, braided, in distributor for veterinary use produced) ....................................................................... 9.3-4801
................................................................................................ 1221 Sodium pertechnetate (99mTc) injection (fission) ............... 1178
Silver, colloidal, for external use........................................... 3552 Sodium pertechnetate (99mTc) injection (non-fission) ....... 1180
Silver nitrate ............................................................................ 3552 Sodium phenylbutyrate.......................................................... 3595
Simeticone ............................................................................... 3553 Sodium phosphate (32P) injection......................................... 1180
Simvastatin .............................................................................. 3554 Sodium picosulfate ................................................................. 3596
Single-dose preparations, uniformity of content (2.9.6.)..... 312 Sodium polystyrene sulfonate ............................................... 3597
Single-dose preparations, uniformity of mass (2.9.5.) ......... 311 Sodium propionate ................................................................. 3598
Sintered-glass filters (2.1.2.) ...................................................... 15 Sodium propyl parahydroxybenzoate .................................. 3599
Sitagliptin phosphate monohydrate .............................. 9.1-4198 Sodium pyrophosphate decahydrate for radiopharmaceutical
Sitagliptin tablets .................................................................... 3557 preparations ................................................................... 9.2-4439
Size-exclusion chromatography (2.2.30.)................................. 47 Sodium risedronate 2.5-hydrate ........................................... 3495
(S)-Lactic acid ......................................................................... 2861 Sodium salicylate .................................................................... 3601
Smallpox vaccine (live) ............................................................ 983 Sodium selenite....................................................................... 3601
Sodium acetate ([1-11C]) injection........................................ 1166 Sodium selenite pentahydrate ............................................... 3602
Sodium acetate trihydrate...................................................... 3558 Sodium (S)-lactate solution................................................... 3587
Sodium alendronate trihydrate ............................................. 3559 Sodium starch glycolate (type A)................................... 9.1-4200
Sodium alginate ...................................................................... 3560 Sodium starch glycolate (type B)................................... 9.1-4202
Sodium aluminium silicate.................................................... 1686 Sodium starch glycolate (type C).......................................... 3604
Sodium amidotrizoate............................................................ 3561 Sodium stearate....................................................................... 3605
Sodium aminosalicylate dihydrate ....................................... 3562 Sodium stearyl fumarate........................................................ 3606
Sodium ascorbate ................................................................... 3563 Sodium sulfate, anhydrous .................................................... 3607
Sodium sulfate decahydrate................................................... 3607 Spheroids and granules, friability of (2.9.41.) ....................... 375
Sodium sulfite ......................................................................... 3608 Spike lavender oil............................................................. 9.5-5617
Sodium sulfite heptahydrate.................................................. 3608 Spiramycin........................................................................ 9.2-4612
Sodium tetrachloroaurate dihydrate for homoeopathic Spirapril hydrochloride monohydrate ................................. 3638
preparations ................................................................... 9.5-5623 Spironolactone ........................................................................ 3639
Sodium thiosulfate.................................................................. 3609 Spot-on preparations....................................................... 9.3-4793
Sodium valproate.................................................................... 3609 Sprays (liquid nasal) and drops (nasal).................................. 867
Soft capsules ..................................................................... 9.4-5287 Sprays, veterinary ............................................................ 9.3-4793
Softening time determination of lipophilic suppositories Squalane................................................................................... 3641
(2.9.22.) .................................................................................... 338 Standard solutions for limit tests (4.1.2.)...................... 9.4-5247
Soft extracts ............................................................................... 817 Standards, reference (5.12.) ............................................ 9.5-5563
Solid dosage forms, dissolution test for (2.9.3.).................... 302 Stannous chloride dihydrate.................................................. 3644
Solid dosage forms, recommendations on dissolution testing Stanozolol ................................................................................ 3644
of (5.17.1.)................................................................................ 761 Staphysagria for homoeopathic preparations ..................... 1612
Solids by mercury porosimetry, porosity and pore-size Star anise.................................................................................. 1529
distribution of (2.9.32.).......................................................... 352 Star anise oil ............................................................................ 1530
Solids, density of (2.2.42.).......................................................... 70 Starches, hydroxyethyl ........................................................... 3649
Solids, gas pycnometric density of (2.9.23.) .......................... 339 Starch glycolate (type A), sodium.................................. 9.1-4200
Solids (porous) including powders, wettability of (2.9.45.).. 381 Starch glycolate (type B), sodium .................................. 9.1-4202
Solifenacin succinate ....................................................... 9.3-4991 Starch glycolate (type C), sodium......................................... 3604
Solubility in alcohol of essential oils (2.8.10.)....................... 284 Starch, hydroxypropyl ............................................................ 3645
Soluble tablets .................................................................. 9.3-4792 Starch, hydroxypropyl, pregelatinised.................................. 3647
Solution calorimetry and microcalorimetry, characterisation Starch, maize ........................................................................... 2969
of crystalline solids by (2.2.61.) ............................................ 109 Starch, pea ............................................................................... 3277
Solutions, emulsions and suspensions, oral ................. 9.3-4786 Starch, potato .......................................................................... 3390
Solutions for haemodialysis................................................... 2628 Starch, pregelatinised ............................................................. 3649
Solutions for haemodialysis, concentrated, water for Starch, rice ............................................................................... 3487
diluting................................................................................... 2627 Starch, wheat ........................................................................... 3937
Solutions for haemofiltration and haemodiafiltration ....... 2631 Starflower (borage) oil, refined ............................................. 1858
Solutions for organ preservation .......................................... 3612 Statistical analysis of results of biological assays and tests
Solutions for peritoneal dialysis............................................ 3299 (5.3.) ................................................................................ 9.2-4353
Solutions, suspensions, intrauterine....................................... 862 Stavudine ................................................................................. 3653
Solvents, residual (5.4.) ................................................... 9.5-5553 Steam sterilisation of aqueous preparations, application of the
Solvents, residual, identification and control (2.4.24.)......... 146 F0 concept (5.1.5.) ................................................................... 580
Somatostatin............................................................................ 3613 Stearic acid............................................................................... 3655
Somatropin ....................................................................... 9.5-5750 Stearoyl macrogolglycerides ........................................... 9.1-4206
Somatropin concentrated solution ................................ 9.5-5752 Stearyl alcohol ......................................................................... 3657
Somatropin for injection ................................................ 9.5-5754 Stem cells, human haematopoietic ....................................... 2682
Somatropin solution for injection ........................................ 3620 Stephania root, fourstamen ................................................... 1357
Sophora flower ........................................................................ 1520 Sterile braided silk suture in distributor for veterinary
Sophora flower-bud................................................................ 1522 use........................................................................................... 1221
Sorbic acid ............................................................................... 3622 Sterile catgut............................................................................ 1207
Sorbitan laurate................................................................ 9.1-4203 Sterile catgut in distributor for veterinary use.................... 1219
Sorbitan oleate.................................................................. 9.1-4203 Sterile containers of plasticised poly(vinyl chloride) for human
Sorbitan palmitate ........................................................... 9.1-4204 blood containing anticoagulant solution (3.2.5.) ............... 432
Sorbitan sesquioleate....................................................... 9.1-4205 Sterile linen thread in distributor for veterinary use ......... 1220
Sorbitan stearate .............................................................. 9.1-4205 Sterile non-absorbable strands in distributor for veterinary
Sorbitan trioleate ............................................................. 9.1-4206 use........................................................................................... 1222
Sorbitol..................................................................................... 3625 Sterile non-absorbable sutures ....................................... 9.5-5603
Sorbitol, liquid (crystallising) ............................................... 3626 Sterile plastic containers for human blood and blood
Sorbitol, liquid (non-crystallising) ....................................... 3627 components (3.2.3.)................................................................ 430
Sorbitol, liquid, partially dehydrated ................................... 3628 Sterile polyamide 6/6 suture in distributor for veterinary
Sotalol hydrochloride............................................................. 3629 use.................................................................................... 9.5-5609
Soya-bean oil, hydrogenated ................................................. 3630 Sterile polyamide 6 suture in distributor for veterinary
Soya-bean oil, refined...................................................... 9.3-4992 use.................................................................................... 9.5-5609
Soya phospholipids for injection ................................... 9.4-5445 Sterile poly(ethylene terephthalate) suture in distributor for
Spanish sage oil ....................................................................... 1524 veterinary use................................................................. 9.5-5609
Specific surface area by air permeability (2.9.14.) ....... 9.1-4053 Sterile products, biological indicators and related microbial
Specific surface area by gas adsorption (2.9.26.) .................. 344 preparations used in the manufacture of (5.1.2.) ...... 9.2-4336
Spectinomycin dihydrochloride pentahydrate.................... 3631 Sterile products, methods of preparation of (5.1.1.) ... 9.2-4333
Spectinomycin sulfate tetrahydrate for veterinary use ...... 3633 Sterile single-use plastic syringes (3.2.8.) .............................. 434
Spectrometry, atomic absorption (2.2.23.).............................. 37 Sterile synthetic absorbable braided sutures ....................... 1212
Spectrometry, atomic emission (2.2.22.) ................................. 36 Sterile synthetic absorbable monofilament sutures ............ 1213
Spectrometry, mass (2.2.43.) ..................................................... 71 Sterility (2.6.1.).......................................................................... 185
Spectrometry, nuclear magnetic resonance (2.2.33.) ............. 54 Sterility, guidelines for using the test for (5.1.9.).................. 592
Spectrometry, X-ray fluorescence (2.2.37.) .................. 9.3-4689 Sterols in fatty oils (2.4.23.) ..................................................... 144
Spectrophotometry, infrared absorption (2.2.24.) ................. 39 Sticks .......................................................................................... 884
Spectrophotometry, ultraviolet and visible absorption Sticks, intrauterine.................................................................... 862
(2.2.25.) ...................................................................................... 41 Sticks, nasal ............................................................................... 868
Spectroscopy, near-infrared (2.2.40.)....................................... 64 St. John’s wort.......................................................................... 1526
Spectroscopy, Raman (2.2.48.).................................................. 86 St. John’s wort dry extract, quantified.................................. 1527
SPF chicken flocks for the production and quality control of Stomata and stomatal index (2.8.3.) ....................................... 283
vaccines (5.2.2.)....................................................................... 599 Stramonium leaf .............................................................. 9.2-4473

Stramonium, prepared .................................................... 9.2-4475 Sweet orange oil ...................................................................... 1535

Strands, sterile non-absorbable, in distributor for veterinary Swelling index (2.8.4.) .............................................................. 284
use .......................................................................................... 1222 Swine erysipelas vaccine (inactivated) ................................. 1104
Streptokinase concentrated solution .................................... 3658 Swine-fever vaccine (live, prepared in cell cultures),
Streptomycin sulfate ............................................................... 3660 classical .................................................................................. 1104
Strontium (89Sr) chloride injection....................................... 1181 Symbols and abbreviations (1.)...................................... 9.2-4275
Strychnos ignatii for homoeopathic preparations....... 9.3-4827 Synthetic absorbable braided sutures, sterile ...................... 1212
Subdivision of tablets ...................................................... 9.3-4790 Synthetic absorbable monofilament sutures, sterile ........... 1213
Sublingual sprays, oromucosal drops and oromucosal Syringes, plastic, sterile single-use (3.2.8.) ............................ 434
sprays............................................................................... 9.3-4787 Syrups................................................................................ 9.3-4787
Sublingual tablets and buccal tablets............................. 9.3-4789 Szechwan lovage rhizome ............................................... 9.4-5314
Substances for pharmaceutical use................................ 9.3-4777
Substances for pharmaceutical use, control of impurities in T
(5.10.) ....................................................................................... 723 Table of physical characteristics of radionuclides mentioned in
Substances of animal origin for the production of the European Pharmacopoeia (5.7.)............................ 9.2-4385
immunological veterinary medicinal products (5.2.5.) ..... 608 Tablets ............................................................................... 9.3-4790
Substitution of in vivo method(s) by in vitro method(s) for the Tablets and capsules, disintegration of (2.9.1.) ..................... 299
quality control of vaccines (5.2.14.) ............................ 9.3-4737 Tablets and powders for rectal solutions and suspensions .. 882
Sub-visible particles, particulate contamination (2.9.19.) ... 335 Tablets, buccal .................................................................. 9.3-4789
Sucralfate .......................................................................... 9.4-5447 Tablets, chewable ............................................................. 9.3-4792
Sucralose ........................................................................... 9.2-4615 Tablets, coated .................................................................. 9.3-4791
Sucrose ..................................................................................... 3664 Tablets, dispersible........................................................... 9.3-4792
Sucrose, liquid.................................................................. 9.4-5448 Tablets, effervescent......................................................... 9.3-4791
Sucrose monopalmitate.......................................................... 3665 Tablets for intrauterine solutions and suspensions .............. 862
Sucrose stearate....................................................................... 3666 Tablets for use in the mouth.................................................... 887
Sufentanil ................................................................................. 3668 Tablets for vaginal solutions and suspensions ...................... 889
Sufentanil citrate ..................................................................... 3669 Tablets, gastro-resistant................................................... 9.3-4791
Sugars, lead in (2.4.10.) ............................................................ 136 Tablets, intrauterine.................................................................. 862
Sugar spheres........................................................................... 3670 Tablets, modified-release ................................................ 9.3-4791
Sulbactam sodium .................................................................. 3671 Tablets, orodispersible..................................................... 9.3-4792
Sulfacetamide sodium ............................................................ 3673 Tablets, resistance to crushing (2.9.8.) ................................... 313
Sulfadiazine ............................................................................. 3674 Tablets, soluble ................................................................. 9.3-4792
Sulfadimethoxine............................................................. 9.4-5450 Tablets, subdivision of..................................................... 9.3-4790
Sulfadimethoxine sodium for veterinary use ............... 9.4-5451 Tablets, sublingual ........................................................... 9.3-4789
Sulfadimidine .......................................................................... 3678 Tablets, uncoated ............................................................. 9.3-4791
Sulfadoxine .............................................................................. 3680 Tablets, uncoated, friability of (2.9.7.) ................................... 312
Sulfafurazole............................................................................ 3681 Tablets, vaginal.......................................................................... 888
Sulfaguanidine ........................................................................ 3681 Tacalcitol monohydrate.......................................................... 3707
Sulfamerazine.......................................................................... 3682 Tacrolimus monohydrate................................................ 9.3-4997
Sulfamethizole......................................................................... 3683 Tadalafil.................................................................................... 3708
Sulfamethoxazole.................................................................... 3684 Talc ........................................................................................... 3710
Sulfamethoxypyridazine for veterinary use......................... 3685 Tamoxifen citrate .................................................................... 3712
Sulfanilamide........................................................................... 3686 Tampons, ear ............................................................................. 856
Sulfasalazine ..................................................................... 9.2-4617 Tampons, medicated ............................................................... 887
Sulfated ash (2.4.14.) ................................................................ 136 Tampons, rectal......................................................................... 882
Sulfates (2.4.13.)........................................................................ 136 Tampons, vaginal, medicated .................................................. 889
Sulfathiazole ............................................................................ 3689 Tamsulosin hydrochloride..................................................... 3714
Sulfinpyrazone......................................................................... 3689 Tannic acid .............................................................................. 3716
Sulfur dioxide (2.5.29.)............................................................. 171 Tannins in herbal drugs (2.8.14.)............................................ 288
Sulfur for external use............................................................ 3691 Tapped density and bulk density of powders (2.9.34.)......... 359
Sulfur for homoeopathic preparations................................. 1614 Tartaric acid............................................................................. 3716
Sulfuric acid............................................................................. 3691 Teat dips............................................................................ 9.3-4793
Sulindac.................................................................................... 3692 Tea tree oil ............................................................................... 1536
Sulpiride................................................................................... 3693 Teat sprays ........................................................................ 9.3-4793
Sultamicillin ............................................................................ 3694 Technetium (99mTc) bicisate injection ........................... 9.2-4440
Sultamicillin tosilate dihydrate ............................................. 3697 Technetium (99mTc) colloidal rhenium sulfide injection ... 1183
Sumatra benzoin ..................................................................... 1273 Technetium (99mTc) colloidal sulfur injection ..................... 1184
Sumatra benzoin tincture ...................................................... 1274 Technetium (99mTc) colloidal tin injection .......................... 1185
Sumatriptan succinate............................................................ 3699 Technetium (99mTc) etifenin injection.................................. 1185
Sunflower oil, refined ............................................................. 3701 Technetium (99mTc) exametazime injection ........................ 1187
Supercritical fluid chromatography (2.2.45.) .......................... 74 Technetium (99mTc) gluconate injection .............................. 1188
Suppositories ............................................................................. 881 Technetium (99mTc) human albumin injection ................... 1189
Suppositories and pessaries, disintegration of (2.9.2.) ......... 301 Technetium (99mTc) macrosalb injection ............................. 1190
Suppositories, lipophilic, softening time determination Technetium (99mTc) mebrofenin injection .................... 9.2-4440
(2.9.22.) .................................................................................... 338 Technetium (99mTc) medronate injection ............................ 1192
Suspensions, solutions and emulsions, oral ................. 9.3-4786 Technetium (99mTc) mertiatide injection ............................. 1193
Suspensions, solutions, intrauterine....................................... 862 Technetium (99mTc) microspheres injection ........................ 1194
Sutures, sterile non-absorbable ...................................... 9.5-5603 Technetium (99mTc) oxidronate injection ............................ 1195
Sutures, sterile synthetic absorbable braided ..................... 1212 Technetium (99mTc) pentetate injection ............................... 1196
Sutures, sterile synthetic absorbable monofilament .......... 1213 Technetium (99mTc) sestamibi injection ........................ 9.2-4441
Suxamethonium chloride ...................................................... 3701 Technetium (99mTc) succimer injection................................ 1199
Suxibuzone .............................................................................. 3702 Technetium (99mTc) tin pyrophosphate injection ............... 1199
Sweet fennel............................................................................. 1353
Teicoplanin .............................................................................. 3717 Tetanus vaccine (adsorbed), assay of (2.7.8.) ........................ 253
Telmisartan .............................................................................. 3718 Tetanus vaccine for veterinary use ....................................... 1106
Temazepam.............................................................................. 3720 Tetracaine hydrochloride....................................................... 3742
Temozolomide.................................................................. 9.4-5455 Tetracosactide.......................................................................... 3743
Tenosynovitis vaccine (live), viral, avian ............................. 1017 Tetracycline ............................................................................. 3744
Tenoxicam ........................................................................ 9.4-5456 Tetracycline hydrochloride.................................................... 3746
Terazosin hydrochloride dihydrate ...................................... 3724 Tetra-O-acetyl-mannose triflate for radiopharmaceutical
Terbinafine hydrochloride ..................................................... 3727 preparations ................................................................... 9.2-4443
Terbutaline sulfate .................................................................. 3728 Tetrazepam .............................................................................. 3747
Terconazole.............................................................................. 3729 Tetryzoline hydrochloride ..................................................... 3749
Terfenadine.............................................................................. 3730 Thallous (201Tl) chloride injection ........................................ 1202
Teriparatide ............................................................................. 3732 Theobromine........................................................................... 3749
Terlipressin ....................................................................... 9.2-4623 Theophylline ........................................................................... 3750
Terminology used in monographs on biological products Theophylline-ethylenediamine ............................................. 3752
(5.2.1.) ...................................................................................... 599 Theophylline-ethylenediamine hydrate ............................... 3754
Test for anticomplementary activity of immunoglobulin Theophylline monohydrate ................................................... 3751
(2.6.17.) ........................................................................... 9.3-4713 Thermal analysis (2.2.34.).......................................................... 57
Test for anti-D antibodies in human immunoglobulin Thermogravimetry (2.2.34.)...................................................... 57
(2.6.26.) .................................................................................... 225 Thiamazole .............................................................................. 3755
Test for aristolochic acids in herbal drugs (2.8.21) .............. 292 Thiamine hydrochloride ........................................................ 3756
Test for extractable volume of parenteral preparations Thiamine nitrate ..................................................................... 3758
(2.9.17.) .................................................................................... 322 Thiamphenicol ........................................................................ 3760
Test for Fc function of immunoglobulin (2.7.9.)......... 9.3-4724 Thin-layer chromatography (2.2.27.) ....................................... 43
Test for methanol and 2-propanol (2.9.11.) .......................... 318 Thiocolchicoside crystallised from ethanol ................. 9.3-4999
Test for neurovirulence of live virus vaccines (2.6.18.)........ 212 Thiocolchicoside hydrate................................................ 9.3-5002
Test for specified micro-organisms (microbiological Thioctic acid............................................................................ 3765
examination of non-sterile products) (2.6.13.) ................... 199 Thiomersal............................................................................... 3766
Testosterone............................................................................. 3734 Thiopental sodium and sodium carbonate ......................... 3767
Testosterone decanoate .......................................................... 3736 Thioridazine ............................................................................ 3768
Testosterone enantate ............................................................. 3738 Thioridazine hydrochloride .................................................. 3770
Testosterone isocaproate ........................................................ 3740 Thomson kudzuvine root ............................................... 9.3-4820
Testosterone propionate......................................................... 3741 Three-lobed sage leaf ............................................................. 1506
Tests for extraneous agents in viral vaccines for human use Threonine ................................................................................ 3771
(2.6.16.) ........................................................................... 9.4-5111 Thyme ...................................................................................... 1538
Tetanus and diphtheria toxins and toxoids, flocculation value Thyme oil, thymol type.......................................................... 1540
(Lf) of, (Ramon assay) (2.7.27.)............................................ 273 Thyme, wild............................................................................. 1559
Tetanus and diphtheria vaccine (adsorbed) .......................... 899 Thymol..................................................................................... 3772
Tetanus and diphtheria vaccine (adsorbed, reduced antigen(s) Thymol type thyme oil........................................................... 1540
content).................................................................................... 900 Tiabendazole ........................................................................... 3773
Tetanus antitoxin for human use .......................................... 1119 Tiamulin for veterinary use................................................... 3774
Tetanus antitoxin for veterinary use..................................... 1123 Tiamulin hydrogen fumarate for veterinary use................. 3777
Tetanus, diphtheria and hepatitis B (rDNA) vaccine Tianeptine sodium.................................................................. 3779
(adsorbed) ............................................................................... 901 Tiapride hydrochloride .......................................................... 3781
Tetanus, diphtheria and pertussis (acellular, component) Tiaprofenic acid ...................................................................... 3782
vaccine (adsorbed) ................................................................. 902 Tibolone................................................................................... 3783
Tetanus, diphtheria and pertussis (acellular, component) Ticarcillin sodium .................................................................. 3784
vaccine (adsorbed, reduced antigen(s) content)................. 903 Tick-borne encephalitis vaccine (inactivated) ...................... 988
Tetanus, diphtheria and pertussis (whole cell) vaccine Ticlopidine hydrochloride ..................................................... 3786
(adsorbed) ............................................................................... 905 Tigecycline........................................................................ 9.4-5457
Tetanus, diphtheria and poliomyelitis (inactivated) vaccine Tilidine hydrochloride hemihydrate .................................... 3788
(adsorbed, reduced antigen(s) content)............................... 906 Timolol maleate ...................................................................... 3789
Tetanus, diphtheria, pertussis (acellular, component) and Tinctures .................................................................................... 817
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5583 Tinidazole ................................................................................ 3791
Tetanus, diphtheria, pertussis (acellular, component) and Tinnevelly senna pods............................................................ 1519
hepatitis B (rDNA) vaccine (adsorbed) ............................... 910 Tinzaparin sodium ................................................................. 3792
Tetanus, diphtheria, pertussis (acellular, component) and Tioconazole ............................................................................. 3793
poliomyelitis (inactivated) vaccine (adsorbed)................... 911 Tiotropium bromide monohydrate ............................... 9.3-5004
Tetanus, diphtheria, pertussis (acellular, component) and Titanium dioxide .................................................................... 3796
poliomyelitis (inactivated) vaccine (adsorbed, reduced Titration, amperometric (2.2.19.) ............................................. 35
antigen(s) content) ................................................................. 913 Titration, potentiometric (2.2.20.)............................................ 35
Tetanus, diphtheria, pertussis (acellular, component), Titrations, complexometric (2.5.11.)...................................... 164
hepatitis B (rDNA), poliomyelitis (inactivated) and Titration, voltametric (2.2.65.)................................................ 112
haemophilus type b conjugate vaccine (adsorbed) ... 9.5-5585 Tizanidine hydrochloride ...................................................... 3797
Tetanus, diphtheria, pertussis (acellular, component), Tobramycin.............................................................................. 3799
poliomyelitis (inactivated) and haemophilus type b conjugate Tocopherol, all-rac-α-............................................................. 3800
vaccine (adsorbed) ........................................................ 9.5-5587 Tocopherol, RRR-α-................................................................ 3801
Tetanus, diphtheria, pertussis (whole cell) and poliomyelitis Tocopheryl acetate, all-rac-α- ............................................... 3802
(inactivated) vaccine (adsorbed) .......................................... 920 α-Tocopheryl acetate concentrate (powder form).............. 3805
Tetanus, diphtheria, pertussis (whole cell), poliomyelitis Tocopheryl acetate, RRR-α-................................................... 3804
(inactivated) and haemophilus type b conjugate vaccine Tocopheryl hydrogen succinate, DL-α- ................................ 3806
(adsorbed) ...................................................................... 9.5-5590 Tocopheryl hydrogen succinate, RRR-α- ............................. 3808
Tetanus immunoglobulin, human ........................................ 2698 Tolbutamide............................................................................. 3810
Tetanus vaccine (adsorbed) ..................................................... 987 Tolfenamic acid ....................................................................... 3811

Tolnaftate ................................................................................. 3812 Tubing used in sets for the transfusion of blood and blood
Tolterodine tartrate................................................................. 3813 components, materials based on plasticised poly(vinyl
Tolu balsam ............................................................................. 1541 chloride) for (3.1.1.2.) ............................................................ 393
Toluenesulfonate (methyl, ethyl and isopropyl) in active Turkey infectious rhinotracheitis vaccine (live) ................. 1107
substances (2.5.40.)................................................................. 179 Turmeric, Javanese.................................................................. 1544
Torasemide .............................................................................. 3815 Turmeric rhizome................................................................... 1545
Tormentil ................................................................................. 1542 Turpentine oil................................................................... 9.4-5316
Tormentil tincture .................................................................. 1542 Tylosin for veterinary use ............................................... 9.3-5008
Tosylchloramide sodium ....................................................... 3816 Tylosin phosphate bulk solution for veterinary use .... 9.3-5012
Total ash (2.4.16.)...................................................................... 137 Tylosin phosphate for veterinary use ............................ 9.3-5017
Total cholesterol in oils rich in omega-3 acids (2.4.32.) ...... 157 Tylosin tartrate for veterinary use ................................. 9.3-5021
Total organic carbon in water for pharmaceutical use Typhoid polysaccharide and hepatitis A (inactivated,
(2.2.44.) ...................................................................................... 73 adsorbed) vaccine ................................................................... 929
Total protein (2.5.33.)............................................................... 172 Typhoid polysaccharide vaccine ............................................. 990
Toxicity, abnormal (2.6.9.)....................................................... 194 Typhoid vaccine ........................................................................ 992
Traditional Chinese medicine, names of herbal drugs used in Typhoid vaccine (live, oral, strain Ty 21a) ............................ 992
(5.22.) .............................................................................. 9.4-5283 Tyrosine ................................................................................... 3870
Tragacanth ............................................................................... 1543 Tyrothricin............................................................................... 3871
Tramadol hydrochloride ........................................................ 3817
Tramazoline hydrochloride monohydrate........................... 3818 U
Trandolapril............................................................................. 3819 Ubidecarenone ........................................................................ 3875
Tranexamic acid...................................................................... 3821 Udder-washes................................................................... 9.3-4793
Transdermal patches ................................................................ 873 Ultraviolet and visible absorption spectrophotometry
Transdermal patches, dissolution test for (2.9.4.)................. 309 (2.2.25.) ...................................................................................... 41
Trapidil..................................................................................... 3822 Ultraviolet ray lamps for analytical purposes (2.1.3.) ............ 15
Trehalose dihydrate ................................................................ 3823 Uncaria stem with hooks ................................................ 9.3-4821
Tretinoin .................................................................................. 3824 Uncoated tablets .............................................................. 9.3-4791
Triacetin ................................................................................... 3825 Undecylenic acid..................................................................... 3876
Triamcinolone ......................................................................... 3826 Uniformity of content of single-dose preparations (2.9.6.).. 312
Triamcinolone acetonide ....................................................... 3827 Uniformity of dosage units (2.9.40.) ............................. 9.1-4055
Triamcinolone hexacetonide ................................................. 3829 Uniformity of dosage units, demonstration using large sample
Triamterene ............................................................................. 3830 sizes (2.9.47.) ........................................................................... 384
Tribenoside.............................................................................. 3832 Uniformity of mass of delivered doses from multidose
Tributyl acetylcitrate............................................................... 3833 containers (2.9.27.) ................................................................. 347
Trichloroacetic acid ................................................................ 3835 Uniformity of mass of single-dose preparations (2.9.5.) ..... 311
Triclabendazole for veterinary use ....................................... 3836 Units of the International System (SI) used in the
Triethanolamine...................................................................... 3849 Pharmacopoeia and equivalence with other units
Triethyl citrate......................................................................... 3837 (1.) ................................................................................... 9.2-4275
Trifluoperazine hydrochloride .............................................. 3837 Unsaponifiable matter (2.5.7.)................................................. 163
Triflusal .................................................................................... 3838 Urea .......................................................................................... 3877
Triglycerides, medium-chain ................................................ 3839 Urofollitropin ................................................................... 9.4-5463
Triglycerides, omega-3-acid .................................................. 3209 Urokinase.......................................................................... 9.4-5464
Triglycerol diisostearate ......................................................... 3841 Uronic acids in polysaccharide vaccines (2.5.22.) ................ 168
Trihexyphenidyl hydrochloride ............................................ 3841 Ursodeoxycholic acid ............................................................ 3880
Trimebutine maleate ....................................................... 9.3-5005 Urtica dioica for homoeopathic preparations..................... 1614
Trimeprazine hemitartrate .................................................... 1666
Trimetazidine dihydrochloride ............................................. 3843
V
Trimethadione......................................................................... 3845
Trimethoprim.......................................................................... 3846 Vaccine component assay by immunonephelometry
Trimipramine maleate............................................................ 3848 (2.7.35.) .................................................................................... 279
Tri-n-butyl phosphate ............................................................ 3834 Vaccines, adsorbed, aluminium in (2.5.13.) .......................... 165
Tritiated (3H) water injection ................................................ 1202 Vaccines, adsorbed, calcium in (2.5.14.)................................ 166
Trolamine................................................................................. 3849 Vaccines and immunosera, phenol in (2.5.15.)..................... 166
Trometamol ............................................................................. 3851 Vaccines and immunosera, veterinary, evaluation of efficacy
Tropicamide............................................................................. 3852 of (5.2.7.).................................................................................. 612
Tropisetron hydrochloride..................................................... 3853 Vaccines and immunosera, veterinary, evaluation of safety
Trospium chloride .................................................................. 3855 (5.2.6.) ...................................................................................... 610
Troxerutin......................................................................... 9.3-5007 Vaccines for human use .................................................. 9.5-5571
Trypsin ..................................................................................... 3857 Vaccines for human use, cell substrates for the production of
Tryptophan .............................................................................. 3858 (5.2.3.) ............................................................................. 9.3-4733
TSE, animal, minimising the risk of transmitting via human Vaccines for human use, conjugated polysaccharide, carrier
and veterinary medicinal products (5.2.8.) ......................... 613 proteins for the production of (5.2.11.) ............................... 626
TSE, animal, products with risk of transmitting agents of.. 832 Vaccines for human use, viral, tests for extraneous agents in
Tuberculin for human use, old.............................................. 3861 (2.6.16.) ........................................................................... 9.4-5111
Tuberculin purified protein derivative, avian ..................... 3862 Vaccines for veterinary use............................................. 9.5-5574
Tuberculin purified protein derivative, bovine ................... 3863 Vaccines for veterinary use, inactivated, healthy chicken flocks
Tuberculin purified protein derivative for human use....... 3864 for the production of (5.2.13.) .............................................. 630
Tuberculosis (BCG) vaccine, freeze-dried............................. 895 Vaccines, polysaccharide, hexosamines in (2.5.20.) ............. 167
Tubes for comparative tests (2.1.5.).......................................... 17 Vaccines, polysaccharide, methylpentoses in (2.5.21.)......... 167
Tubing and closures, silicone elastomer for (3.1.9.) ............. 409 Vaccines, polysaccharide, nucleic acids in (2.5.17.) ............. 166
Tubing and containers for total parenteral nutrition Vaccines, polysaccharide, O-acetyl in (2.5.19.) ..................... 167
preparations, poly(ethylene - vinyl acetate) for Vaccines, polysaccharide, phosphorus in (2.5.18.) ............... 166
(3.1.7.) ............................................................................. 9.2-4318 Vaccines, polysaccharide, protein in (2.5.16.)....................... 166
Vaccines, polysaccharide, ribose in (2.5.31.)......................... 171 Vinpocetine ............................................................................. 3914

Vaccines, polysaccharide, sialic acid in (2.5.23.) .................. 168 Viper venom antiserum, European ...................................... 1119
Vaccines, polysaccharide, uronic acids in (2.5.22.) .............. 168 Viral diarrhoea vaccine (inactivated), bovine ..................... 1023
Vaccines, SPF chicken flocks for the production and quality Viral hepatitis type I vaccine (live), duck ............................ 1047
control of (5.2.2.) ................................................................... 599 Viral rhinotracheitis vaccine (inactivated), feline .............. 1059
Vaccines, substitution of in vivo method(s) by in vitro Viral rhinotracheitis vaccine (live), feline ........................... 1060
method(s) for the quality control of (5.2.14.) ............ 9.3-4737 Viral safety (5.1.7.).................................................................... 591
Vaccines, veterinary, cell cultures for the production of Viral tenosynovitis vaccine (live), avian .............................. 1017
(5.2.4.) ...................................................................................... 606 Viral vaccines for human use, tests for extraneous agents in
Vaccines, viral live, test for neurovirulence (2.6.18.) ........... 212 (2.6.16.) ........................................................................... 9.4-5111
Vaginal capsules ........................................................................ 888 Viscometer method, capillary (2.2.9.)...................................... 27
Vaginal foams............................................................................ 889 Viscometer methods, falling ball and automatic rolling ball
Vaginal preparations ................................................................ 887 (2.2.49.) ........................................................................... 9.3-4690
Vaginal preparations, semi-solid ............................................ 889 Viscose wadding, absorbent .................................................. 3915
Vaginal solutions and suspensions, tablets for...................... 889 Viscosity (2.2.8.)............................................................... 9.4-5099
Vaginal solutions, emulsions and suspensions...................... 888 Viscosity - rotating viscometer method (2.2.10.) ................... 28
Vaginal tablets ........................................................................... 888 Visible and ultraviolet absorption spectrophotometry
Vaginal tampons, medicated ................................................... 889 (2.2.25.) ...................................................................................... 41
Valaciclovir hydrochloride ............................................. 9.3-5029 Visible particles, particulate contamination (2.9.20.) .......... 337
Valaciclovir hydrochloride, hydrated ............................ 9.3-5032 Vitamin A ................................................................................ 3917
Valerian dry aqueous extract ............................................... 1549 Vitamin A concentrate (oily form), synthetic .............. 9.3-5036
Valerian dry hydroalcoholic extract ..................................... 1550 Vitamin A concentrate (powder form), synthetic ....... 9.3-5037
Valerian root..................................................................... 9.1-4111 Vitamin A concentrate (solubilisate/emulsion),
Valerian root, cut ............................................................. 9.1-4113 synthetic.......................................................................... 9.3-5038
Valerian tincture ..................................................................... 1554 Voltametric titration (2.2.65.) ................................................. 112
Validation of nucleic acid amplification techniques for the Volumetric analysis (4.2.) ............................................... 9.4-5258
detection of B19 virus (B19V) DNA in plasma pools : Volumetric solutions (4.2.2.) .......................................... 9.4-5258
guidelines................................................................................. 214 Volumetric solutions (4.2.2.) .......................................... 9.5-5550
Validation of nucleic acid amplification techniques for the Volumetric solutions, primary standards for (4.2.1.).. 9.4-5258
detection of hepatitis C virus (HCV) RNA in plasma pools : von Willebrand factor, human.............................................. 2701
guidelines................................................................................. 214 von Willebrand factor, human, assay of (2.7.21.) ................. 268
Valine ....................................................................................... 3890 Voriconazole............................................................................ 3921
Valnemulin hydrochloride for veterinary use .................... 3892
Valproate, sodium................................................................... 3609 W
Valproic acid............................................................................ 3893 Warfarin sodium..................................................................... 3927
Valsartan .................................................................................. 3895 Warfarin sodium clathrate..................................................... 3928
Vancomycin hydrochloride ................................................... 3896 Washes, nasal............................................................................. 868
Vanillin..................................................................................... 3898 Water (15O) injection .............................................................. 1203
Vapour, preparations to be converted into................... 9.4-5289 Water, determination by distillation (2.2.13.) ......................... 31
Vardenafil hydrochloride trihydrate..................................... 3898 Water for diluting concentrated haemodialysis solutions.. 2627
Varicella immunoglobulin for intravenous administration, Water for injections ......................................................... 9.1-4215
human .................................................................................... 2700 Water for pharmaceutical use, total organic carbon in
Varicella immunoglobulin, human ...................................... 2700 (2.2.44.) ...................................................................................... 73
Varicella, measles, mumps and rubella vaccine (live).......... 953 Water for preparation of extracts.......................................... 3932
Varicella vaccine (live) ............................................................. 994 Water, highly purified ............................................................ 3933
Vectors for human use, adenovirus ........................................ 742 Water in essential oils (2.8.5.) ................................................. 284
Vectors for human use, plasmid ............................................. 740 Water in gases (2.5.28.) ............................................................ 170
Vectors for human use, plasmid, bacterial cells used for the Water : micro determination (2.5.32.) ........................... 9.4-5107
manufacture of........................................................................ 741 Water, purified ................................................................. 9.4-5475
Vectors for human use, poxvirus ............................................ 744 Water : semi-micro determination (2.5.12.) ................. 9.4-5107
Vecuronium bromide ...................................................... 9.1-4211 Water-solid interactions: determination of sorption-
Vedaprofen for veterinary use............................................... 3901 desorption isotherms and of water activity (2.9.39) .......... 369
Vegetable fatty oils .................................................................... 848 Wettability of porous solids including powders (2.9.45.) .... 381
Venlafaxine hydrochloride .................................................... 3902 Wheat-germ oil, refined......................................................... 3937
Verapamil hydrochloride ....................................................... 3904 Wheat-germ oil, virgin........................................................... 3938
Verbena herb ........................................................................... 1555 Wheat starch............................................................................ 3937
Veterinary liquid preparations for cutaneous White beeswax ........................................................................ 1804
application ...................................................................... 9.3-4792 White horehound ................................................................... 1557
Veterinary medicinal products, immunological, substances of White peony root.................................................................... 1472
animal origin for the production of (5.2.5.)........................ 608 White soft paraffin.................................................................. 3270
Veterinary semi-solid preparations for oral use ................... 890 Wild pansy (flowering aerial parts)...................................... 1558
Veterinary vaccines and immunosera, evaluation of efficacy of Wild thyme.............................................................................. 1559
(5.2.7.) ...................................................................................... 612 Willow bark ............................................................................. 1561
Viability, nucleated cell count and (2.7.29.) .......................... 275 Willow bark dry extract ......................................................... 1562
Vibriosis (cold-water) vaccine (inactivated) for Wool alcohols .......................................................................... 3938
salmonids........................................................................ 9.2-4428 Wool fat.................................................................................... 3939
Vibriosis vaccine (inactivated) for salmonids .............. 9.2-4429 Wool fat, hydrogenated.......................................................... 3943
VICH (5.8.)....................................................................... 9.4-5265 Wool fat, hydrous.................................................................... 3944
Vigabatrin ................................................................................ 3906 Wormwood.............................................................................. 1563
Vinblastine sulfate .................................................................. 3907
Vincristine sulfate................................................................... 3908
X
Vindesine sulfate.............................................................. 9.3-5034
Vinorelbine tartrate ......................................................... 9.4-5469 Xanthan gum.................................................................... 9.2-4627

Xenon (133Xe) injection .......................................................... 1204 Zanthoxylum bungeanum pericarp...................................... 1565

X-ray fluorescence spectrometry (2.2.37.).................... 9.3-4689 Zidovudine .............................................................................. 3962
X-ray powder diffraction (XRPD), characterisation of Zinc acetate dihydrate ............................................................ 3964
crystalline and partially crystalline solids by (2.9.33.)....... 354 Zinc acexamate ....................................................................... 3965
Xylazine hydrochloride for veterinary use .......................... 3948 Zinc chloride ........................................................................... 3967
Xylitol ....................................................................................... 3949 Zinc gluconate......................................................................... 3967
Xylometazoline hydrochloride.............................................. 3951 Zinc oxide ................................................................................ 3968
Xylose ....................................................................................... 3953 Zinc stearate ............................................................................ 3968
Zinc sulfate heptahydrate ...................................................... 3969
Y Zinc sulfate hexahydrate ........................................................ 3970
Yarrow ...................................................................................... 1564 Zinc sulfate monohydrate...................................................... 3970
Yeasticidal, bactericidal or fungicidal activity of antiseptic Zinc undecylenate .................................................................. 3970
medicinal products, determination of (5.1.11.) ......... 9.2-4348 Ziprasidone hydrochloride monohydrate............................ 3971
Yellow beeswax........................................................................ 1805 Ziprasidone mesilate trihydrate ............................................ 3973
Yellow fever vaccine (live) ....................................................... 996 Zolmitriptan ..................................................................... 9.5-5759
Yellow soft paraffin................................................................. 3271 Zolpidem tartrate.................................................................... 3975
Yersiniosis vaccine (inactivated) for salmonids ........... 9.2-4430 Zopiclone ................................................................................. 3976
Yohimbine hydrochloride ...................................................... 3957 Zoster (shingles) vaccine (live), herpes.................................. 982
Zuclopenthixol decanoate...................................................... 3978
Z
Zanamivir hydrate .................................................................. 3961
Numerics Acidum pipemidicum trihydricum........................................ 3346

α-1-Proteinasi inhibitor humanum ...................................... 2694 Acidum salicylicum................................................................. 3528
Acidum (S)-lacticum............................................................... 2861
A Acidum sorbicum .................................................................... 3622
Acidum stearicum ................................................................... 3655
Abacaviri sulfas ....................................................................... 1619
Acidum succinicum ad praeparationes homoeopathicas...... 9.5-
Absinthii herba ........................................................................ 1563
5621
Acaciae gummi ........................................................................ 1229
Acidum sulfuricum ................................................................. 3691
Acaciae gummi dispersione desiccatum ................................ 1620
Acidum tartaricum ................................................................. 3716
Acamprosatum calcicum ........................................................ 1621
Acidum thiocticum.................................................................. 3765
Acanthopanacis gracilistyli cortex .................................. 9.2-4447
Acidum tiaprofenicum............................................................ 3782
Acarbosum ............................................................................... 1622
Acidum tolfenamicum ............................................................ 3811
Acari ad producta allergenica................................................ 3077
Acidum tranexamicum........................................................... 3821
Acebutololi hydrochloridum................................................... 1624
Acidum trichloroaceticum...................................................... 3835
Aceclofenacum.................................................................. 9.3-4835
Acidum undecylenicum .......................................................... 3876
Acemetacinum ......................................................................... 1628
Acidum ursodeoxycholicum ................................................... 3880
Acesulfamum kalicum ............................................................ 1629
Acidum valproicum ................................................................ 3893
Acetazolamidum ..................................................................... 1630
Acitretinum....................................................................... 9.5-5627
Acetonum ................................................................................. 1632
Adapalenum ............................................................................ 1646
Acetylcholini chloridum.......................................................... 1633
Adeninum ................................................................................ 1647
Acetylcysteinum....................................................................... 1634
Adenosinum............................................................................. 1648
β-Acetyldigoxinum.................................................................. 1635
Adeps A 3-O-desacyl-4-monophosphorylatus ..................... 2207
Acetylenum (1 per centum) in nitrogenio intermixtum ....... 9.3-
Adeps lanae.............................................................................. 3939
4836
Adeps lanae cum aqua............................................................ 3944
Aciclovirum.............................................................................. 1642
Adeps lanae hydrogenatus...................................................... 3943
Acidi methacrylici et ethylis acrylatis polymerisati 1:1 dispersio
Adeps solidus ........................................................................... 2639
30 per centum ........................................................................ 3017
Adeps solidus cum additamentis ........................................... 2640
Acidi methacrylici et ethylis acrylatis polymerisatum 1:1 ... 3015
Adrenalini tartras ................................................................... 1652
Acidi methacrylici et methylis methacrylatis polymerisatum
Adrenalinum ........................................................................... 1650
1:1 ........................................................................................... 3018
Aer medicinalis........................................................................ 1653
Acidi methacrylici et methylis methacrylatis polymerisatum
Aer medicinalis artificiosus .................................................... 1655
1:2 ........................................................................................... 3019
Aether ....................................................................................... 2421
Acidum 4-aminobenzoicum................................................... 1702
Aether anaestheticus ............................................................... 2422
Acidum aceticum glaciale....................................................... 1632
Aetherolea .................................................................................. 814
Acidum acetylsalicylicum ....................................................... 1638
Agar .......................................................................................... 1230
Acidum adipicum.................................................................... 1649
Agni casti fructus..................................................................... 1231
Acidum alginicum................................................................... 1665
Agni casti fructus extractum siccum ..................................... 1232
Acidum amidotrizoicum dihydricum ................................... 1694
Agrimoniae herba ................................................................... 1233
Acidum aminocaproicum....................................................... 1703
Akebiae caulis .......................................................................... 1234
Acidum ascorbicum ......................................................... 9.3-4843
Alaninum ................................................................................. 1656
Acidum asparticum.......................................................... 9.3-4845
Albendazolum................................................................... 9.4-5321
Acidum benzoicum ................................................................. 1818
Albumini humani solutio ....................................................... 2660
Acidum boricum...................................................................... 1859
Alchemillae herba.................................................................... 1235
Acidum caprylicum ................................................................. 1938
Alcohol 2,4-dichlorobenzylicus .............................................. 2242
Acidum chenodeoxycholicum................................................. 2026
Alcohol benzylicus ............................................................ 9.4-5327
Acidum citricum...................................................................... 2098
Alcohol cetylicus ...................................................................... 2023
Acidum citricum monohydricum .......................................... 2099
Alcohol cetylicus et stearylicus ............................................... 2019
Acidum edeticum .................................................................... 2344
Alcohol cetylicus et stearylicus emulsificans A ..................... 2019
Acidum etacrynicum............................................................... 2413
Alcohol cetylicus et stearylicus emulsificans B ..................... 2021
Acidum folicum hydricum .............................................. 9.5-5662
Alcoholes adipis lanae............................................................. 3938
Acidum formicum ............................................................ 9.4-5376
Alcohol isopropylicus .............................................................. 2819
Acidum fusidicum ................................................................... 2552
Alcohol oleicus ......................................................................... 3199
Acidum glutamicum ............................................................... 2593
Alcohol stearylicus................................................................... 3657
Acidum hydrochloridum concentratum................................ 2704
Alcuronii chloridum................................................................ 1658
Acidum hydrochloridum dilutum ......................................... 2704
Alfacalcidolum......................................................................... 1660
Acidum iopanoicum................................................................ 2797
Alfadexum......................................................................... 9.3-4838
Acidum ioxaglicum ................................................................. 2803
Alfentanili hydrochloridum ................................................... 1662
Acidum lacticum ..................................................................... 2860
Alfuzosini hydrochloridum ............................................. 9.1-4123
Acidum lactobionicum ........................................................... 2863
Alimemazini hemitartras ....................................................... 1666
Acidum maleicum ................................................................... 2971
Allantoinum............................................................................. 1667
Acidum malicum..................................................................... 2972
Allii sativi bulbi pulvis............................................................ 1364
Acidum medronicum ad radiopharmaceutica .............. 9.2-4437
Allium sativum ad praeparationes homoeopathicas............ 1590
Acidum mefenamicum ........................................................... 2987
Allopurinolum ......................................................................... 1668
Acidum nalidixicum ............................................................... 3115
Almagatum .............................................................................. 1670
Acidum nicotinicum ........................................................ 9.3-4961
Aloe barbadensis ..................................................................... 1236
Acidum niflumicum................................................................ 3152
Aloe capensis............................................................................ 1237
Acidum nitricum..................................................................... 3161
Aloes extractum siccum normatum....................................... 1238
Acidum oleicum ...................................................................... 3197
Alovudini (18F) solutio iniectabilis......................................... 1127
Acidum oxolinicum................................................................. 3237
Alprazolamum......................................................................... 1672
Acidum palmiticum ................................................................ 3259
Alprenololi hydrochloridum................................................... 1674
Acidum phosphoricum concentratum ................................... 3333
Alprostadilum.......................................................................... 1675
Acidum phosphoricum dilutum............................................. 3334
Alteplasum ad iniectabile ....................................................... 1677
Acidum picricum ad praeparationes homoeopathicas ........ 1587
Althaeae folium ....................................................................... 1429
Althaeae radix ......................................................................... 1430 Aprotininum ............................................................................ 1747

Altizidum ................................................................................. 1681 Aqua ad dilutionem solutionum concentratarum ad
Alumen..................................................................................... 1682 haemodialysim ...................................................................... 2627
Aluminii chloridum hexahydricum....................................... 1682 Aqua ad extracta praeparanda.............................................. 3932
Aluminii hydroxidum hydricum ad adsorptionem ............. 1682 Aqua ad iniectabile .......................................................... 9.1-4215
Aluminii magnesii silicas................................................. 9.3-4839 Aquae (15O) solutio iniectabilis .............................................. 1203
Aluminii natrii silicas ............................................................. 1686 Aquae tritiatae (3H) solutio iniectabilis ................................ 1202
Aluminii oxidum hydricum ................................................... 1684 Aqua purificata ................................................................ 9.4-5475
Aluminii phosphas hydricus................................................... 1686 Aqua valde purificata ............................................................. 3933
Aluminii phosphatis liquamen............................................... 1685 Arachidis oleum hydrogenatum............................................. 1752
Aluminii stearas ...................................................................... 1687 Arachidis oleum raffinatum ................................................... 1752
Aluminii sulfas ........................................................................ 1689 Argenti nitras........................................................................... 3552
Alverini citras ................................................................... 9.3-4841 Argentum colloidale ad usum externum............................... 3552
Amanita phalloides ad praeparationes homoeopathicas...... 9.5- Arginini aspartas..................................................................... 1754
5621 Arginini hydrochloridum ....................................................... 1755
Amantadini hydrochloridum ................................................. 1691 Argininum................................................................................ 1753
Ambroxoli hydrochloridum............................................. 9.2-4499 Argon ........................................................................................ 1756
Amfetamini sulfas ................................................................... 1694 Aripiprazolum ......................................................................... 1757
Amikacini sulfas ...................................................................... 1698 Arnicae flos .............................................................................. 1251
Amikacinum ............................................................................ 1696 Arnicae tinctura ...................................................................... 1253
Amiloridi hydrochloridum dihydricum ......................... 9.2-4500 Arsenii trioxidum ad praeparationes homoeopathicas.. 9.5-5623
Aminoglutethimidum ............................................................. 1704 Articaini hydrochloridum ...................................................... 1758
Amiodaroni hydrochloridum ................................................. 1706 Ascorbylis palmitas ................................................................. 1762
Amisulpridum ......................................................................... 1707 Asparaginum monohydricum ................................................ 1762
Amitriptylini hydrochloridum ............................................... 1709 Aspartamum............................................................................ 1763
Amlodipini besilas................................................................... 1710 Astragali mongholici radix .............................................. 9.2-4449
Ammoniae (13N) solutio iniectabilis ...................................... 1129 Atenololum ....................................................................... 9.1-4124
Ammoniae solutio concentrata .............................................. 1712 Atomoxetini hydrochloridum ................................................ 1767
Ammonii bromidum ............................................................... 1714 Atorvastatinum calcicum trihydricum.................................. 1769
Ammonii carbonas ad praeparationes homoeopathicas ...... 9.3- Atovaquonum.......................................................................... 1771
4827 Atractylodis lanceae rhizoma................................................. 1259
Ammonii chloridum ............................................................... 1715 Atractylodis macrocephalae rhizoma .................................... 1260
Ammonii glycyrrhizas............................................................. 1716 Atracurii besilas....................................................................... 1772
Ammonii hydrogenocarbonas................................................ 1717 Atropa belladonna ad praeparationes homoeopathicas ....... 9.2-
Ammonio methacrylatis copolymerum A ............................. 1712 4492
Ammonio methacrylatis copolymerum B ............................. 1713 Atropini sulfas .................................................................. 9.3-4849
Amobarbitalum ....................................................................... 1717 Atropinum ........................................................................ 9.3-4847
Amobarbitalum natricum ...................................................... 1718 Aucklandiae radix............................................................ 9.2-4450
Amomi fructus......................................................................... 1239 Aurantii amari epicarpii et mesocarpii tinctura.................. 1284
Amomi fructus rotundus ........................................................ 1502 Aurantii amari epicarpium et mesocarpium ....................... 1283
Amorolfini hydrochloridum ................................................... 1719 Aurantii amari flos ................................................................. 1284
Amoxicillinum natricum ........................................................ 1720 Aurantii dulcis aetheroleum .................................................. 1535
Amoxicillinum trihydricum ................................................... 1723 Auricularia ................................................................................ 855
Amphotericinum B.................................................................. 1725 Azaperonum ad usum veterinarium..................................... 1778
Ampicillinum........................................................................... 1727 Azathioprinum ........................................................................ 1779
Ampicillinum natricum .......................................................... 1729 Azelastini hydrochloridum..................................................... 1780
Ampicillinum trihydricum ..................................................... 1731 Azithromycinum .............................................................. 9.3-4850
Amygdalae oleum raffinatum ................................................ 1671
Amygdalae oleum virginale.................................................... 1671 B
Amyla hydroxyethyla.............................................................. 3649 Bacampicillini hydrochloridum ............................................. 1787
Amylmetacresolum.................................................................. 1733 Bacitracinum .................................................................... 9.1-4129
Amylum hydroxypropylum .................................................... 3645 Bacitracinum zincum....................................................... 9.1-4133
Amylum hydroxypropylum pregelificatum........................... 3647 Baclofenum .............................................................................. 1794
Amylum pregelificatum .......................................................... 3649 Ballotae nigrae herba .............................................................. 1289
Anamirta cocculus ad praeparationes homoeopathicas ...... 1597 Balsamum peruvianum .......................................................... 1479
Anastrozolum ................................................................... 9.3-4842 Balsamum tolutanum............................................................. 1541
Andrographidis herba ...................................................... 9.3-4807 Bambuteroli hydrochloridum ................................................ 1795
Anemarrhenae asphodeloides rhizoma ................................. 1241 Barbitalum............................................................................... 1797
Angelicae archangelicae radix................................................ 1242 Barii chloridum dihydricum ad praeparationes
Angelicae dahuricae radix............................................... 9.3-4808 homoeopathicas ..................................................................... 1594
Angelicae pubescentis radix............................................. 9.3-4810 Barii sulfas ............................................................................... 1797
Angelicae sinensis radix................................................... 9.1-4100 BCG ad immunocurationem.................................................... 894
Anisi aetheroleum ................................................................... 1248 Beclometasoni dipropionas..................................................... 1799
Anisi fructus...................................................................... 9.2-4448 Beclometasoni dipropionas monohydricus ........................... 1802
Anisi stellati aetheroleum ....................................................... 1530 Belamcandae chinensis rhizoma............................................ 1266
Anisi stellati fructus ................................................................ 1529 Belladonnae folii extractum siccum normatum............ 9.2-4453
Antazolini hydrochloridum.................................................... 1737 Belladonnae folii tinctura normata................................ 9.2-4454
Anticorpora monoclonalia ad usum humanum .................... 826 Belladonnae folium .......................................................... 9.2-4452
Antithrombinum III humanum densatum........................... 2664 Belladonnae pulvis normatus.......................................... 9.2-4456
Apis mellifera ad praeparationes homoeopathicas............... 1592 Benazeprili hydrochloridum .................................................. 1806
Apomorphini hydrochloridum hemihydricum..................... 1744 Bendroflumethiazidum........................................................... 1807
Aprepitantum .......................................................................... 1746 Benperidolum .......................................................................... 1808
Aprotinini solutio concentrata ............................................... 1749

Benserazidi hydrochloridum .................................................. 1810 Cadmii sulfas hydricus ad praeparationes

Bentonitum .............................................................................. 1811homoeopathicas ..................................................................... 1596
Benzalkonii chloridi solutio.................................................... 1813 Calcifediolum........................................................................... 1901
Benzalkonii chloridum ........................................................... 1811 Calcii acetas ............................................................................. 1910
Benzbromaronum ................................................................... 1815 Calcii ascorbas ......................................................................... 1911
Benzethonii chloridum............................................................ 1816 Calcii carbonas ........................................................................ 1912
Benzocainum .................................................................... 9.4-5326 Calcii chloridum dihydricum................................................. 1912
Benzoe sumatranus ................................................................. 1273 Calcii chloridum hexahydricum ............................................ 1913
Benzoe tonkinensis .................................................................. 1272 Calcii dobesilas monohydricus............................................... 1914
Benzois sumatrani tinctura .................................................... 1274 Calcii fluoridum ad praeparationes homoeopathicas... 9.5-5624
Benzois tonkinensis tinctura .................................................. 1274 Calcii folinas ............................................................................ 1914
Benzoylis peroxidum cum aqua............................................. 1819 Calcii glucoheptonas ............................................................... 1917
Benzylis benzoas...................................................................... 1822 Calcii gluconas......................................................................... 1917
Benzylpenicillinum benzathinum .......................................... 1822 Calcii gluconas ad iniectabile................................................. 1919
Benzylpenicillinum kalicum............................................ 9.2-4505 Calcii gluconas anhydricus..................................................... 1918
Benzylpenicillinum natricum.......................................... 9.2-4508 Calcii glycerophosphas ............................................................ 1920
Benzylpenicillinum procainum ....................................... 9.2-4506 Calcii hydrogenophosphas ...................................................... 1921
Betacarotenum ........................................................................ 1829 Calcii hydrogenophosphas dihydricus ................................... 1922
Betadexum ........................................................................ 9.3-4857 Calcii hydroxidum .................................................................. 1923
Betahistini dihydrochloridum ................................................ 1831 Calcii iodidum tetrahydricum ad praeparationes
Betahistini mesilas................................................................... 1832 homoeopathicas ..................................................................... 1596
Betamethasoni acetas.............................................................. 1835 Calcii lactas.............................................................................. 1923
Betamethasoni dipropionas............................................. 9.3-4858 Calcii lactas monohydricus .................................................... 1924
Betamethasoni natrii phosphas.............................................. 1839 Calcii lactas pentahydricus .................................................... 1924
Betamethasoni valeras ............................................................ 1840 Calcii lactas trihydricus.......................................................... 1925
Betamethasonum..................................................................... 1833 Calcii laevulinas dihydricus ................................................... 1928
Betaxololi hydrochloridum..................................................... 1842 Calcii levofolinas pentahydricus ............................................ 1926
Betulae folium ......................................................................... 1276Calcii pantothenas .................................................................. 1929
Bezafibratum ........................................................................... 1843Calcii stearas............................................................................ 1930
Bicalutamidum........................................................................ 1845 Calcii sulfas dihydricus........................................................... 1932
Bifonazolum............................................................................. 1846Calcipotriolum......................................................................... 1902
Biotinum ........................................................................... 9.5-5631Calcipotriolum monohydricum ............................................. 1904
Biperideni hydrochloridum .................................................... 1848 Calcitoninum salmonis........................................................... 1906
Bisacodylum............................................................................. 1850Calcitriolum............................................................................. 1909
Bismuthi subcarbonas............................................................. 1851 Calendulae flos ........................................................................ 1297
Bismuthi subgallas .................................................................. 1852 Camelliae sinensis non fermentata folia ........................ 9.4-5304
Bismuthi subnitras ponderosus .............................................. 1853 Camphora racemica................................................................ 1934
Bismuthi subsalicylas .............................................................. 1854 Candesartanum cilexetili ....................................................... 1935
Bisoprololi fumaras ................................................................. 1855 Capecitabinum ........................................................................ 1937
Bistortae rhizoma .................................................................... 1278 Capsici extractum spissum normatum.................................. 1303
Bleomycini sulfas..................................................................... 1857 Capsici fructus ......................................................................... 1300
Boldi folii extractum siccum................................................... 1293 Capsici oleoresina raffinata et normata................................ 1302
Boldi folium ............................................................................. 1292
Capsici tinctura normata ....................................................... 1304
Boraginis officinalis oleum raffinatum ................................. 1858 Capsulae............................................................................ 9.4-5287
Captoprilum ............................................................................ 1940
Borax ........................................................................................ 1859
Brimonidini tartras................................................................. 1864 Carbacholum ........................................................................... 1942
Bromazepamum ...................................................................... 1865 Carbamazepinum.................................................................... 1943
Bromhexini hydrochloridum........................................... 9.1-4137 Carbasalatum calcicum .......................................................... 1945
Bromocriptini mesilas ............................................................. 1868 Carbidopum...................................................................... 9.4-5333
Bromperidoli decanoas ........................................................... 1872 Carbimazolum......................................................................... 1948
Bromperidolum ....................................................................... 1870 Carbo activatus ....................................................................... 2025
Brompheniramini maleas ...................................................... 1873 Carbocisteinum ....................................................................... 1949
Brotizolamum.......................................................................... 1875 Carbomera ............................................................................... 1950
Budesonidum ........................................................................... 1876 Carbonei dioxidum ................................................................. 1951
Bufexamacum.......................................................................... 1878 Carbonei monoxidum............................................................. 1953
Buflomedili hydrochloridum .................................................. 1879 Carbonei monoxidum (15O) ................................................... 1130
Bumetanidum.......................................................................... 1881 Carbonei monoxidum (5 per centum) in nitrogenio
Bupivacaini hydrochloridum ................................................. 1882 intermixtum.................................................................... 9.3-4865
Bupleuri radix................................................................... 9.4-5303 Carboplatinum ........................................................................ 1954
Buprenorphini hydrochloridum............................................. 1886 Carboprostum trometamolum ............................................... 1955
Buprenorphinum..................................................................... 1884 Carboxymethylamylum natricum A .............................. 9.1-4200
Buserelinum ............................................................................. 1887
Carboxymethylamylum natricum B............................... 9.1-4202
Buspironi hydrochloridum ..................................................... 1889 Carboxymethylamylum natricum C ..................................... 3604
Busulfanum.............................................................................. 1891Carisoprodolum....................................................................... 1956
Butylhydroxyanisolum............................................................ 1893 Carmellosum............................................................................ 1957
Butylhydroxytoluenum ........................................................... 1894 Carmellosum calcicum .................................................... 9.4-5335
Butylis parahydroxybenzoas .................................................. 1892 Carmellosum natricum........................................................... 1958
Carmellosum natricum conexum .......................................... 2174
C Carmellosum natricum substitutum humile ........................ 1958
C1-esterasi inhibitor humanus............................................... 2665 Carmustinum .......................................................................... 1960
Cabergolinum .......................................................................... 1897 Carprofenum ad usum veterinarium.................................... 1961
Carrageenanum....................................................................... 1962
Carteololi hydrochloridum..................................................... 1963
Carthami flos ........................................................................... 1503 Chlorpromazini hydrochloridum .......................................... 2050

Carthami oleum raffinatum................................................... 3523 Chlorpropamidum .................................................................. 2051
Carvedilolum ........................................................................... 1965 Chlorprothixeni hydrochloridum .......................................... 2052
Carvi aetheroleum .................................................................. 1306 Chlortalidonum....................................................................... 2054
Carvi fructus............................................................................ 1305 Chlortetracyclini hydrochloridum ......................................... 2055
Caryophylli floris aetheroleum............................................... 1323 Cholecalciferoli pulvis ............................................................. 2061
Caryophylli flos........................................................................ 1322 Cholecalciferolum.................................................................... 2058
Cefaclorum............................................................................... 1969 Cholecalciferolum densatum oleosum................................... 2059
Cefadroxilum monohydricum................................................ 1970 Cholecalciferolum in aqua dispergibile ................................. 2062
Cefalexinum monohydricum ................................................. 1972 Cholesterolum.......................................................................... 2064
Cefalotinum natricum ............................................................ 1973 Cholesterolum ad usum parenteralem ........................... 9.2-4517
Cefamandoli nafas .................................................................. 1974 Cholini ([11C]methyl) solutio iniectabilis ....................... 9.4-5297
Cefapirinum natricum............................................................ 1976 Chondroitini natrii sulfas....................................................... 2067
Cefatrizinum propylen glycolum ........................................... 1977 Chorda resorbilis sterilis ......................................................... 1207
Cefazolinum natricum............................................................ 1978 Chorda resorbilis sterilis in fuso ad usum veterinarium ..... 1219
Cefepimi dihydrochloridum monohydricum................. 9.2-4513 Chromii (51Cr) edetatis solutio iniectabilis ........................... 1131
Cefiximum ........................................................................ 9.4-5335 Chymotrypsinum..................................................................... 2069
Cefoperazonum natricum....................................................... 1984 Ciclesonidum ........................................................................... 2070
Cefotaximum natricum .......................................................... 1985 Ciclopirox olaminum .............................................................. 2072
Cefoxitinum natricum ............................................................ 1987 Ciclopiroxum ........................................................................... 2071
Cefpodoximum proxetili......................................................... 1989 Ciclosporinum ......................................................................... 2074
Cefprozilum monohydricum ........................................... 9.4-5337 Cilastatinum natricum ........................................................... 2075
Cefradinum.............................................................................. 1994 Cilazaprilum............................................................................ 2077
Ceftazidimum pentahydricum ........................................ 9.3-4865 Cimetidini hydrochloridum............................................. 9.4-5345
Ceftazidimum pentahydricum et natrii carbonas ad Cimetidinum..................................................................... 9.4-5343
iniectabile........................................................................ 9.3-4867 Cimicifugae rhizoma............................................................... 1285
Ceftriaxonum natricum.......................................................... 2000 Cinchocaini hydrochloridum ................................................. 2082
Cefuroximum axetili ........................................................ 9.2-4515 Cinchonae cortex..................................................................... 1314
Cefuroximum natricum.......................................................... 2003 Cinchonae extractum fluidum normatum............................ 1316
Celecoxibum............................................................................. 2004 Cineolum.................................................................................. 2083
Celiprololi hydrochloridum .................................................... 2005 Cinnamomi cassiae aetheroleum ........................................... 1310
Cellulae stirpes haematopoieticae humanae ........................ 2682 Cinnamomi cortex .................................................................. 1317
Cellulosi acetas ................................................................. 9.2-4516 Cinnamomi zeylanici corticis aetheroleum .......................... 1318
Cellulosi acetas butyras........................................................... 2008 Cinnamomi zeylanici folii aetheroleum................................ 1318
Cellulosi acetas phthalas......................................................... 2009 Cinnarizinum .......................................................................... 2084
Cellulosi pulvis......................................................................... 2013 Ciprofibratum.......................................................................... 2085
Cellulosum microcristallinum................................................ 2010 Ciprofloxacini hydrochloridum ............................................. 2088
Cellulosum microcristallinum et carmellosum natricum.... 3068 Ciprofloxacinum...................................................................... 2086
Centaurii herba ....................................................................... 1310 Cisatracurii besilas ........................................................... 9.4-5347
Centellae asiaticae herba ........................................................ 1311 Cisplatinum ............................................................................. 2094
Cera alba .................................................................................. 1804 Citaloprami hydrobromidum.......................................... 9.3-4876
Cera carnauba ......................................................................... 1960 Citaloprami hydrochloridum .......................................... 9.3-4877
Cera flava ................................................................................. 1805 Citri reticulatae aetheroleum ................................................. 1428
Cetirizini dihydrochloridum .................................................. 2017 Citri reticulatae epicarpium et mesocarpium....................... 1426
Cetobemidoni hydrochloridum .............................................. 2848 Citronellae aetheroleum ......................................................... 1319
Cetostearylis isononanoas ...................................................... 2022 Cladribinum ............................................................................ 2100
Cetrimidum ............................................................................. 2022 Clarithromycinum ........................................................... 9.4-5351
Cetylis palmitas ....................................................................... 2024 Clazurilum ad usum veterinarium ....................................... 2104
Cetylpyridinii chloridum ........................................................ 2025 Clebopridi malas ..................................................................... 2106
Chamomillae romanae flos .................................................... 1313 Clemastini fumaras.......................................................... 9.3-4879
Chelidonii herba...................................................................... 1380 Clematidis armandii caulis ............................................. 9.3-4813
Chinidini sulfas ....................................................................... 3461 Clenbuteroli hydrochloridum................................................. 2109
Chinini hydrochloridum......................................................... 3463 Clindamycini hydrochloridum........................................ 9.3-4880
Chinini sulfas........................................................................... 3464 Clindamycini phosphas .......................................................... 2111
Chitosani hydrochloridum ..................................................... 2028 Clioquinolum........................................................................... 2114
Chlorali hydras ........................................................................ 2029 Clobazamum ........................................................................... 2115
Chlorambucilum ..................................................................... 2029 Clobetasoli propionas.............................................................. 2116
Chloramphenicoli natrii succinas.......................................... 2033 Clobetasoni butyras................................................................. 2118
Chloramphenicoli palmitas .................................................... 2032 Clofaziminum.......................................................................... 2120
Chloramphenicolum ........................................................ 9.1-4142 Clofibratum.............................................................................. 2122
Chlorcyclizini hydrochloridum .............................................. 2034 Clomifeni citras ................................................................ 9.5-5635
Chlordiazepoxidi hydrochloridum ........................................ 2036 Clomipramini hydrochloridum.............................................. 2124
Chlordiazepoxidum................................................................. 2035 Clonazepamum ....................................................................... 2126
Chlorhexidini diacetas ..................................................... 9.4-5339 Clonidini hydrochloridum...................................................... 2127
Chlorhexidini digluconatis solutio.................................. 9.3-4871 Clopamidum ............................................................................ 2128
Chlorhexidini dihydrochloridum.................................... 9.4-5341 Clopidogreli besilas.................................................................. 2129
Chlormadinoni acetas............................................................. 2043 Clopidogreli hydrochloridum ................................................. 2131
Chlorobutanolum.................................................................... 2045 Clopidogreli hydrogenosulfas ................................................. 2133
Chlorobutanolum hemihydricum.......................................... 2046 Closantelum natricum dihydricum ad usum
Chlorocresolum........................................................................ 2046 veterinarium .......................................................................... 2134
Chloroquini phosphas ............................................................. 2047 Clotrimazolum ........................................................................ 2136
Chloroquini sulfas ................................................................... 2048 Cloxacillinum natricum ......................................................... 2137
Chlorphenamini maleas ......................................................... 2048 Clozapinum ............................................................................. 2139

Cocaini hydrochloridum......................................................... 2140 Delphinium staphisagria ad praeparationes

Cocois oleum raffinatum ........................................................ 2141 homoeopathicas ..................................................................... 1612
Cocoylis caprylocapras ............................................................ 2142 Dembrexini hydrochloridum monohydricum ad usum
Codeini hydrochloridum dihydricum............................. 9.5-5636 veterinarium ................................................................... 9.2-4526
Codeini phosphas hemihydricus ..................................... 9.5-5641 Demeclocyclini hydrochloridum ............................................ 2203
Codeini phosphas sesquihydricus........................................... 2148 Deptropini citras...................................................................... 2205
Codeinum monohydricum .............................................. 9.5-5639 Dequalinii chloridum.............................................................. 2206
Codergocrini mesilas ............................................................... 2149 Desfluranum ............................................................................ 2209
Codonopsidis radix........................................................... 9.3-4815 Desipramini hydrochloridum................................................. 2210
Coffeinum ................................................................................ 1898 Deslanosidum .......................................................................... 2211
Coffeinum monohydricum ..................................................... 1899 Desloratadinum....................................................................... 2212
Coicis semen...................................................................... 9.2-4457 Desmopressinum ..................................................................... 2213
Colae semen ............................................................................. 1326 Desogestrelum.......................................................................... 2215
Colchicinum...................................................................... 9.4-5354 Desoxycortoni acetas............................................................... 2216
Colestyraminum ...................................................................... 2164 Detomidini hydrochloridum ad usum veterinarium........... 2217
Colistimethatum natricum .............................................. 9.4-5356 Dexamethasoni acetas ............................................................ 2220
Colistini sulfas ......................................................................... 2166 Dexamethasoni isonicotinas................................................... 2222
Colophonium ........................................................................... 1327 Dexamethasoni natrii phosphas ............................................ 2223
Compressi .......................................................................... 9.3-4790 Dexamethasonum ................................................................... 2218
Copolymerum macrogolo et alcoholi poly(vinylico) Dexchlorpheniramini maleas................................................. 2226
constatum............................................................................... 2945 Dexpanthenolum..................................................................... 2227
Copolymerum methacrylatis butylati basicum.............. 9.4-5325 Dextranomerum ...................................................................... 2231
Copovidonum ................................................................... 9.3-4882 Dextranum 1 ad iniectabile ............................................ 9.4-5363
Coptidis rhizoma .............................................................. 9.1-4102 Dextranum 40 ad iniectabile.................................................. 2229
Coriandri aetheroleum ........................................................... 1330 Dextranum 60 ad iniectabile.................................................. 2230
Coriandri fructus..................................................................... 1329 Dextranum 70 ad iniectabile.................................................. 2231
Corpora ad usum pharmaceuticum ............................... 9.3-4777 Dextrinum......................................................................... 9.2-4527
Cortisoni acetas ....................................................................... 2170 Dextromethorphani hydrobromidum ................................... 2233
Crataegi folii cum flore extractum fluidum quantificatum.. 1387 Dextromoramidi tartras ......................................................... 2234
Crataegi folii cum flore extractum siccum ............................ 1386 Dextropropoxypheni hydrochloridum................................... 2235
Crataegi folium cum flore....................................................... 1385 Diacereinum ............................................................................ 2236
Crataegi fructus ....................................................................... 1384 Diazepamum ........................................................................... 2238
Cresolum crudum.................................................................... 2173 Diazoxidum ............................................................................. 2240
Croci sativi stigma ad praeparationes homoeopathicas ...... 1599 Dibrompropamidini diisetionas............................................. 2240
Crospovidonum ....................................................................... 2175 Dibutylis phthalas ................................................................... 2241
Crotamitonum......................................................................... 2177 Diclazurilum ad usum veterinarium .................................... 2243
Cupri acetas monohydricus ad praeparationes Diclofenacum kalicum ............................................................ 2245
homoeopathicas ..................................................................... 1599 Diclofenacum natricum.......................................................... 2246
Cupri sulfas.............................................................................. 2169 Dicloxacillinum natricum ...................................................... 2248
Cupri sulfas pentahydricus..................................................... 2170 Dicycloverini hydrochloridum ........................................ 9.1-4147
Cupri tetramibi tetrafluoroboras ad radiopharmaceutica ... 9.2- Didanosinum ........................................................................... 2250
4435 Dienogestum ............................................................................ 2252
Cuprum ad praeparationes homoeopathicas........................ 1600 Diethylcarbamazini citras ...................................................... 2254
Curcumae longae rhizoma ..................................................... 1545 Diethylenglycoli aether monoethylicus.................................. 2256
Curcumae zanthorrhizae rhizoma ........................................ 1544 Diethylenglycoli palmitostearas ............................................. 2257
Cyamopsidis seminis pulvis.................................................... 1381 Diethylis phthalas.................................................................... 2253
Cyanocobalamini (57Co) capsulae ......................................... 1132 Diethylstilbestrolum ................................................................ 2258
Cyanocobalamini (57Co) solutio ............................................ 1133 Difloxacini hydrochloridum trihydricum ad usum
Cyanocobalamini (58Co) capsulae ......................................... 1134 veterinarium .......................................................................... 2259
Cyanocobalamini (58Co) solutio ............................................ 1134 Digitalis purpureae folium ..................................................... 1336
Cyanocobalaminum................................................................ 2178 Digitoxinum............................................................................. 2260
Cyclizini hydrochloridum....................................................... 2179 Digoxinum ............................................................................... 2261
Cyclopentolati hydrochloridum ............................................. 2180 Dihydralazini sulfas hydricus ......................................... 9.2-4528
Cyclophosphamidum .............................................................. 2181 Dihydrocodeini hydrogenotartras.......................................... 2266
Cynarae folii extractum siccum ...................................... 9.4-5302 Dihydroergocristini mesilas.................................................... 2267
Cynarae folium................................................................. 9.4-5301 Dihydroergotamini mesilas .................................................... 2269
Cyproheptadini hydrochloridum ........................................... 2182 Dihydroergotamini tartras ..................................................... 2271
Cyproteroni acetas .................................................................. 2183 Dihydrostreptomycini sulfas ad usum veterinarium ........... 2272
Cysteini hydrochloridum monohydricum............................. 2185 Dihydrotachysterolum ............................................................ 2274
Cystinum .................................................................................. 2187 Dikalii clorazepas .................................................................... 2291
Cytarabinum............................................................................ 2188 Dikalii phosphas...................................................................... 2292
Diltiazemi hydrochloridum.................................................... 2276
D Dimenhydrinatum .................................................................. 2277
Dacarbazinum......................................................................... 2193 Dimercaprolum ....................................................................... 2279
Dalteparinum natricum ......................................................... 2194 Dimethylacetamidum ............................................................. 2280
Danaparoidum natricum ....................................................... 2196 Dimethylis sulfoxidum............................................................ 2279
Dapsonum................................................................................ 2198 Dimeticonum ........................................................................... 2281
Daunorubicini hydrochloridum............................................. 2199 Dimetindeni maleas ................................................................ 2282
D-Camphora ............................................................................ 1932 Dinatrii clodronas tetrahydricus ........................................... 2119
Decylis oleas ............................................................................. 2200 Dinatrii edetas ......................................................................... 2297
Deferipronum ................................................................... 9.5-5647 Dinatrii etidronas.................................................................... 2433
Deferoxamini mesilas ...................................................... 9.3-4887 Dinatrii pamidronas pentahydricus...................................... 3259
Dinatrii phosphas.................................................................... 2298
Dinatrii phosphas dihydricus................................................. 2299 Epirubicini hydrochloridum................................................... 2365

Dinatrii phosphas dodecahydricus ........................................ 2299 Eplerenonum............................................................................ 2367
Dinitrogenii oxidum ............................................................... 3166 Equiseti herba .......................................................................... 1347
Dinoprostonum ................................................................ 9.2-4529 Ergocalciferolum...................................................................... 2368
Dinoprostum trometamolum ................................................. 2283 Ergometrini maleas ................................................................. 2370
Dioscoreae nipponicae rhizoma...................................... 9.5-5613 Ergotamini tartras................................................................... 2371
Dioscoreae oppositifoliae rhizoma......................................... 1337 Erythritolum ............................................................................ 2373
Diosminum .............................................................................. 2286 Erythromycini estolas.............................................................. 2378
Diphenhydramini hydrochloridum ....................................... 2288 Erythromycini ethylsuccinas ........................................... 9.3-4893
Diphenoxylati hydrochloridum.............................................. 2289 Erythromycini lactobionas .............................................. 9.2-4536
Dipivefrini hydrochloridum ................................................... 2290 Erythromycini stearas ............................................................. 2388
Diprophyllinum ....................................................................... 2293 Erythromycinum ..................................................................... 2374
Dipyridamolum....................................................................... 2294 Erythropoietini solutio concentrata ....................................... 2391
Dirithromycinum .................................................................... 2296 Escitaloprami oxalas ............................................................... 2397
Disopyramidi phosphas .......................................................... 2301 Escitalopramum ...................................................................... 2395
Disopyramidum....................................................................... 2300 Eserini salicylas ....................................................................... 3335
Disulfiramum .......................................................................... 2302 Esketamini hydrochloridum............................................ 9.3-4895
Dithranolum ............................................................................ 2303 Esomeprazolum magnesicum dihydricum ............................ 2401
DL-Methioninum ..................................................................... 3025 Esomeprazolum magnesicum trihydricum ........................... 2402
DL-α-Tocopherylis hydrogenosuccinas .................................. 3806 Estradioli benzoas ................................................................... 2404
Dobutamini hydrochloridum................................................. 2304 Estradioli valeras ..................................................................... 2407
Docetaxelum ............................................................................ 2305 Estradiolum hemihydricum ............................................ 9.1-4151
Docetaxelum trihydricum ...................................................... 2307 Estriolum........................................................................... 9.5-5651
Dodecylis gallas ....................................................................... 2309 Estrogeni coniuncti.................................................................. 2410
Domperidoni maleas............................................................... 2312 Etamsylatum............................................................................ 2414
Domperidonum ....................................................................... 2310 Etanerceptum.................................................................... 9.5-5652
Dopamini hydrochloridum .................................................... 2313 Ethacridini lactas monohydricus ........................................... 2415
Dopexamini dihydrochloridum ............................................. 2315 Ethambutoli hydrochloridum ................................................ 2416
Dorzolamidi hydrochloridum ................................................ 2316 Ethanolum (96 per centum) ................................................... 2417
Dosulepini hydrochloridum ............................................ 9.4-5364 Ethanolum anhydricum ......................................................... 2419
Doxaprami hydrochloridum .................................................. 2319 Ethinylestradiolum .................................................................. 2422
Doxazosini mesilas.................................................................. 2320 Ethionamidum......................................................................... 2424
Doxepini hydrochloridum ...................................................... 2322 Ethosuximidum ................................................................ 9.4-5369
Doxorubicini hydrochloridum ............................................... 2323 Ethylcellulosum................................................................. 9.2-4539
Doxycyclini hyclas ................................................................... 2324 Ethylendiaminum.................................................................... 2431
Doxycyclinum monohydricum............................................... 2326 Ethylenglycoli monopalmitostearas ....................................... 2430
Doxylamini hydrogenosuccinas ............................................. 2328 Ethylis acetas............................................................................ 2427
Droperidolum .......................................................................... 2329 Ethylis oleas ............................................................................. 2428
Drospirenonum ....................................................................... 2331 Ethylis parahydroxybenzoas................................................... 2428
Drynariae rhizoma........................................................... 9.2-4459 Ethylis parahydroxybenzoas natricus.................................... 3577
Duloxetini hydrochloridum.................................................... 2332 Ethylmorphini hydrochloridum ............................................. 2431
Dutasteridum........................................................................... 2334 Etilefrini hydrochloridum....................................................... 2433
Dydrogesteronum .................................................................... 2336 Etodolacum .............................................................................. 2435
Etofenamatum ......................................................................... 2437
E Etomidatum ............................................................................. 2439
Ebastinum ................................................................................ 2341 Etoposidum ....................................................................... 9.1-4152
Echinaceae angustifoliae radix............................................... 1447 Eucalypti aetheroleum ............................................................ 1349
Echinaceae pallidae radix....................................................... 1467 Eucalypti folium ...................................................................... 1349
Echinaceae purpureae herba .................................................. 1486 Eucommiae cortex............................................................ 9.2-4462
Echinaceae purpureae radix................................................... 1488 Eugenolum ............................................................................... 2443
Ecliptae herba ................................................................... 9.2-4461 Evodiae fructus ................................................................. 9.2-4464
Econazoli nitras....................................................................... 2343 Exemestanum .......................................................................... 2445
Econazolum ............................................................................. 2342
Edrophonii chloridum............................................................. 2345 F
Eleutherococci radix................................................................ 1344 Factor humanus von Willebrandi.......................................... 2701
Emedastini difumaras............................................................. 2346 Factoris IX coagulationis humani (ADNr) pulvis ad solutionem
Emetini hydrochloridum pentahydricum ............................. 2347 iniectabilem .................................................................... 9.3-4920
Emplastra transcutanea............................................................ 873 Factoris IX coagulationis humani (ADNr) solutio
Enalaprilatum dihydricum..................................................... 2349 concentrata ..................................................................... 9.3-4915
Enalaprili maleas .................................................................... 2348 Factoris VIIa coagulationis humani (ADNr) solutio
Enilconazolum ad usum veterinarium ................................. 2351 concentrata ..................................................................... 9.5-5687
Enoxaparinum natricum........................................................ 2352 Factor IX coagulationis humanus.......................................... 2674
Enoxolonum............................................................................. 2354 Factor VII coagulationis humanus ........................................ 2666
Enrofloxacinum ad usum veterinarium ........................ 9.2-4535 Factor VIII coagulationis humanus....................................... 2672
Entacaponum........................................................................... 2357 Factor VIII coagulationis humanus (ADNr) ........................ 2673
Entecavirum monohydricum ................................................. 2359 Factor XI coagulationis humanus.......................................... 2681
Ephedrae herba ....................................................................... 1346 Fagopyri herba......................................................................... 1294
Ephedrini hydrochloridum ..................................................... 2362 Famotidinum ........................................................................... 2449
Ephedrini racemici hydrochloridum ..................................... 2363 Febantelum ad usum veterinarium....................................... 2450
Ephedrinum ............................................................................. 2360 Felbinacum .............................................................................. 2451
Ephedrinum hemihydricum ................................................... 2361 Felodipinum............................................................................. 2452
Epinastini hydrochloridum .................................................... 2364 Felypressinum .......................................................................... 2454

Fenbendazolum ad usum veterinarium................................ 2455 Fluvoxamini maleas................................................................ 2523

Fenbufenum ............................................................................. 2456 Foeniculi amari fructus .......................................................... 1352
Fenofibratum ........................................................................... 2457 Foeniculi amari fructus aetheroleum .................................... 1279
Fenoteroli hydrobromidum .................................................... 2458 Foeniculi amari herbae aetheroleum .................................... 1280
Fentanyli citras ................................................................. 9.4-5374 Foeniculi dulcis fructus........................................................... 1353
Fentanylum....................................................................... 9.4-5373 Follitropini solutio concentrata....................................... 9.5-5669
Fenticonazoli nitras ................................................................ 2462 Follitropinum.................................................................... 9.5-5663
Ferri chloridum hexahydricum.............................................. 2463 Formaldehydi solutio (35 per centum).................................. 2537
Ferrosi fumaras ....................................................................... 2464 Formoteroli fumaras dihydricus ............................................ 2537
Ferrosi gluconas....................................................................... 2465 Foscarnetum natricum hexahydricum........................... 9.2-4546
Ferrosi sulfas desiccatus .......................................................... 2466 Fosfomycinum calcicum ......................................................... 2541
Ferrosi sulfas heptahydricus ................................................... 2467 Fosfomycinum natricum ........................................................ 2542
Ferrum ad praeparationes homoeopathicas ......................... 1601 Fosfomycinum trometamolum............................................... 2543
Fexofenadini hydrochloridum................................................ 2468 Fosinoprilum natricum........................................................... 2544
Fibrini glutinum ...................................................................... 2469 Fragmenta epithelii phaneraeque bestiarum ad producta
Fibrinogenum humanum ....................................................... 2682 allergenica .............................................................................. 1736
Fila non resorbilia sterilia ............................................... 9.5-5603 Framycetini sulfas ................................................................... 2547
Fila non resorbilia sterilia in fuso ad usum veterinarium .. 1222 Frangulae cortex...................................................................... 1358
Fila resorbilia synthetica monofilamenta sterilia................. 1213 Frangulae corticis extractum siccum normatum ................. 1359
Fila resorbilia synthetica torta sterilia .................................. 1212 Fraxini folium.......................................................................... 1257
Filgrastimi solutio concentrata........................................ 9.3-4899 Fraxini rhynchophyllae cortex ............................................... 1360
Filipendulae ulmariae herba.................................................. 1436 Fructosum ................................................................................ 2548
Filum bombycis tortum sterile in fuso ad usum Fucus vel Ascophyllum............................................................ 1405
veterinarium .......................................................................... 1221 Fulvestrantum ......................................................................... 2549
Filum ethyleni polyterephthalici sterile in fuso ad usum Fumariae herba ................................................................ 9.2-4465
veterinarium ................................................................... 9.5-5609 Furosemidum.................................................................... 9.4-5377
Filum lini sterile in fuso ad usum veterinarium .................. 1220
Filum polyamidicum-6/6 sterile in fuso ad usum G
veterinarium ................................................................... 9.5-5609 Gabapentinum.................................................................. 9.4-5381
Filum polyamidicum-6 sterile in fuso ad usum Gadobutrolum monohydricum....................................... 9.3-4905
veterinarium ................................................................... 9.5-5609 Gadodiamidum hydricum............................................... 9.1-4165
Finasteridum............................................................................ 2473 Galactosum .............................................................................. 2562
Fipronilum ad usum veterinarium................................. 9.5-5661 Galantamini hydrobromidum ............................................... 2563
Flavoxati hydrochloridum...................................................... 2476 Gallii (67Ga) citratis solutio iniectabilis................................. 1148
Flecainidi acetas ...................................................................... 2478 Gallii (68Ga) chloridi solutio ad radio-signandum............... 1148
Flubendazolum........................................................................ 2479 Gallii (68Ga) edotreotidi solutio iniectabilis.......................... 1150
Flucloxacillinum magnesicum octahydricum....................... 2480 Gammadexum.................................................................. 9.4-5382
Flucloxacillinum natricum..................................................... 2482 Ganciclovirum ......................................................................... 2566
Fluconazolum .......................................................................... 2484 Gardeniae fructus.................................................................... 1299
Flucytosinum ........................................................................... 2485 Gefitinibum....................................................................... 9.3-4906
Fludarabini phosphas ............................................................. 2487 Gelatina............................................................................. 9.3-4907
Fludeoxyglucosi (18F) solutio iniectabilis............................... 1135 Gemcitabini hydrochloridum................................................. 2570
Fludrocortisoni acetas............................................................. 2489 Gemfibrozilum.................................................................. 9.5-5679
Flumazenili (N-[11C]methyl) solutio iniectabilis.................. 1137 Gentamicini sulfas................................................................... 2573
Flumazenilum.......................................................................... 2490 Gentianae radix....................................................................... 1365
Flumequinum .......................................................................... 2492 Gentianae tinctura .................................................................. 1366
Flumetasoni pivalas ................................................................ 2493 Gestodenum ............................................................................. 2575
Flunarizini dihydrochloridum ............................................... 2494 Ginkgonis extractum siccum raffinatum et quantificatum.. 1368
Flunitrazepamum.................................................................... 2495 Ginkgonis folium ..................................................................... 1370
Flunixini megluminum ad usum veterinarium ................... 2496 Ginseng extractum siccum...................................................... 1374
Fluocinoloni acetonidum........................................................ 2497 Ginseng radix........................................................................... 1372
Fluocortoloni pivalas .............................................................. 2500 Glibenclamidum...................................................................... 2577
Fluoresceinum ......................................................................... 2501 Gliclazidum.............................................................................. 2578
Fluoresceinum natricum......................................................... 2502 Glimepiridum .......................................................................... 2580
Fluoridi (18F) solutio ad radio-signandum ........................... 1139 Glipizidum ............................................................................... 2582
Fluorocholini (18F) solutio iniectabilis................................... 1139 Glossa ................................................................................ 9.2-4423
Fluorodopae (18F) ab electrophila substitutione solutio Glucagonum humanum.......................................................... 2584
iniectabilis .............................................................................. 1141 Glucosamini hydrochloridum ......................................... 9.5-5680
Fluoroethyl-L-tyrosini (18F) solutio iniectabilis.............. 9.3-4797 Glucosamini sulfas kalii chloridum................................ 9.5-5681
Fluoromisonidazoli (18F) solutio iniectabilis......................... 1146 Glucosamini sulfas natrii chloridum.............................. 9.5-5682
Fluorouracilum................................................................. 9.2-4545 Glucosum .......................................................................... 9.1-4167
Fluoxetini hydrochloridum .................................................... 2505 Glucosum liquidum................................................................. 2590
Flupentixoli dihydrochloridum.............................................. 2507 Glucosum liquidum dispersione desiccatum......................... 2591
Fluphenazini decanoas ........................................................... 2509 Glucosum monohydricum ............................................... 9.1-4168
Fluphenazini dihydrochloridum............................................ 2510 Glutathionum .......................................................................... 2594
Fluphenazini enantas ............................................................. 2512 Glycerol-formalum .................................................................. 2600
Flurazepami monohydrochloridum ...................................... 2513 Glyceroli dibehenas ................................................................. 2598
Flurbiprofenum ....................................................................... 2514 Glyceroli distearas ................................................................... 2599
Fluspirilenum .......................................................................... 2516 Glyceroli monocaprylas .......................................................... 2601
Flutamidum ............................................................................. 2517 Glyceroli monocaprylocapras ................................................. 2602
Fluticasoni propionas.............................................................. 2518 Glyceroli monolinoleas .......................................................... 2603
Flutrimazolum......................................................................... 2520 Glyceroli mono-oleas............................................................... 2604
Fluvastatinum natricum......................................................... 2521
Glyceroli monostearas 40-55 ........................................... 9.2-4551 Hydroxycarbamidum.............................................................. 2720

Glyceroli trinitratis solutio ..................................................... 2606 Hydroxychloroquini sulfas .............................................. 9.3-4922
Glycerolum............................................................................... 2595 Hydroxyethylcellulosum .................................................. 9.3-4924
Glycerolum (85 per centum) .................................................. 2597 Hydroxyethylis salicylas.......................................................... 2723
Glycinum.................................................................................. 2608 Hydroxypropylbetadexum...................................................... 2725
Glycopyrronii bromidum........................................................ 2609 Hydroxypropylcellulosum....................................................... 2727
Gonadorelini acetas ......................................................... 9.4-5383 Hydroxypropylcellulosum substitutum humile ............. 9.5-5691
Gonadotrophinum chorionicum..................................... 9.4-5385 Hydroxyzini hydrochloridum ......................................... 9.5-5693
Gonadotropinum sericum equinum ad usum Hymecromonum...................................................................... 2731
veterinarium .......................................................................... 2613 Hymenopteri venena ad producta allergenica ..................... 2732
Goserelinum............................................................................. 2614 Hyoscini butylbromidum................................................. 9.5-5694
Gossypii oleum hydrogenatum............................................... 2173 Hyoscini hydrobromidum ...................................................... 2735
Gramicidinum ......................................................................... 2616 Hyoscinum ............................................................................... 2733
Graminis rhizoma ................................................................... 1331 Hyoscyamini sulfas ................................................................. 2737
Granisetroni hydrochloridum ................................................ 2617 Hyoscyamus niger ad praeparationes homoeopathicas ....... 1604
Granula ad praeparationes homoeopathicas........................ 1586 Hyperici herba ......................................................................... 1526
Granula homoeopathica imbuta..................................... 9.1-4118 Hyperici herbae extractum siccum quantificatum............... 1527
Granula homoeopathica velata.............................................. 1587 Hypericum perforatum ad praeparationes
Granulata................................................................................... 860 homoeopathicas ..................................................................... 1605
Griseofulvinum........................................................................ 2619 Hypromellosi phthalas ............................................................ 2740
Guaiacolum ............................................................................. 2620 Hypromellosum ....................................................................... 2738
Guaifenesinum.................................................................. 9.4-5385
Guanethidini monosulfas ....................................................... 2623 I
Guaranae semen............................................................... 9.4-5306 Ibuprofenum ............................................................................ 2745
Guar galactomannanum ........................................................ 2623 Ichthammolum ........................................................................ 2747
Idoxuridinum .......................................................................... 2748
H Iecoris aselli domestici oleum................................................. 2151
Halofantrini hydrochloridum ................................................ 2633 Iecoris aselli oleum A .............................................................. 2155
Haloperidoli decanoas ............................................................ 2636 Iecoris aselli oleum B .............................................................. 2159
Haloperidolum ........................................................................ 2634 Ifosfamidum............................................................................. 2749
Halothanum............................................................................. 2637 Imatinibi mesilas .............................................................. 9.2-4561
Hamamelidis cortex ................................................................ 1382 Imipenemum monohydricum ......................................... 9.4-5391
Hamamelidis folium ............................................................... 1383 Imipramini hydrochloridum .................................................. 2754
Harpagophyti extractum siccum............................................ 1334 Immunoglobulinum anti-T lymphocytorum ex animali ad
Harpagophyti radix................................................................. 1335 usum humanum.................................................................... 1741
Hederae folium ........................................................................ 1401 Immunoglobulinum humanum anti-D ................................ 2662
Hedera helix ad praeparationes homoeopathicas ................ 1602 Immunoglobulinum humanum anti-D ad usum
Helianthi annui oleum raffinatum........................................ 3701 intravenosum......................................................................... 2663
Helium...................................................................................... 2642 Immunoglobulinum humanum hepatitidis A ...................... 2683
Heparina massae molecularis minoris.................................. 2646 Immunoglobulinum humanum hepatitidis B ...................... 2684
Heparinum calcicum ....................................................... 9.3-4911 Immunoglobulinum humanum hepatitidis B ad usum
Heparinum natricum....................................................... 9.3-4913 intravenosum......................................................................... 2684
Heptaminoli hydrochloridum ................................................ 2649 Immunoglobulinum humanum morbillicum ....................... 2685
Hexamidini diisetionas........................................................... 2650 Immunoglobulinum humanum normale ad usum
Hexetidinum ............................................................................ 2651 intramusculum ...................................................................... 2685
Hexylresorcinolum .................................................................. 2652 Immunoglobulinum humanum normale ad usum
Hibisci sabdariffae flos............................................................ 1498 intravenosum......................................................................... 2687
Hippocastani semen ................................................................ 1389 Immunoglobulinum humanum normale ad usum
Hippocastani seminis extractum siccum normatum............ 1390 subdermicum ......................................................................... 2689
Histamini dihydrochloridum ................................................. 2653 Immunoglobulinum humanum rabicum.............................. 2696
Histaminum ad praeparationes homoeopathicas.......... 9.1-4118 Immunoglobulinum humanum rubellae .............................. 2698
Histidini hydrochloridum monohydricum............................ 2655 Immunoglobulinum humanum tetanicum........................... 2698
Histidinum ............................................................................... 2654 Immunoglobulinum humanum varicellae............................ 2700
Homatropini hydrobromidum ............................................... 2657 Immunoglobulinum humanum varicellae ad usum
Homatropini methylbromidum ............................................. 2658 intravenosum......................................................................... 2700
Houttuyniae herba ........................................................... 9.4-5307 Immunosera ad usum veterinarium ....................................... 823
Hyaluronidasum ..................................................................... 2702 Immunosera ex animale ad usum humanum ........................ 821
Hydralazini hydrochloridum .......................................... 9.2-4557 Immunoserum botulinicum ................................................... 1115
Hydrargyri dichloridum ......................................................... 3003 Immunoserum contra venena viperarum europaearum..... 1119
Hydrastis canadensis ad praeparationes homoeopathicas .. 1603 Immunoserum diphthericum ................................................. 1115
Hydrastis rhizoma................................................................... 1378 Immunoserum gangraenicum (Clostridium novyi) ............. 1116
Hydrochlorothiazidum ........................................................... 2705 Immunoserum gangraenicum (Clostridium perfringens) ... 1117
Hydrocodoni hydrogenotartras 2.5-hydricus ....................... 2706 Immunoserum gangraenicum (Clostridium septicum) ....... 1118
Hydrocortisoni acetas ............................................................. 2711 Immunoserum gangraenicum mixtum ................................. 1116
Hydrocortisoni hydrogenosuccinas........................................ 2713 Immunoserum tetanicum ad usum humanum.................... 1119
Hydrocortisonum .................................................................... 2708 Immunoserum tetanicum ad usum veterinarium ............... 1123
Hydrogenii peroxidum 30 per centum .................................. 2715 Indapamidum.......................................................................... 2755
Hydrogenii peroxidum 3 per centum .................................... 2715 Indii (111In) chloridi solutio .................................................... 1153
Hydromorphoni hydrochloridum .......................................... 2716 Indii (111In) oxini solutio ........................................................ 1154
Hydroxocobalamini acetas..................................................... 2717 Indii (111In) pentetatis solutio iniectabilis ............................. 1154
Hydroxocobalamini chloridum.............................................. 2718 Indinaviri sulfas ...................................................................... 2757
Hydroxocobalamini sulfas...................................................... 2719 Indometacinum ....................................................................... 2759

Inhalanda.......................................................................... 9.4-5288 Kalii citras................................................................................ 3378

Insulini zinci amorphi suspensio iniectabilis........................ 2779 Kalii clavulanas ....................................................................... 3378
Insulini zinci cristallini suspensio iniectabilis ...................... 2779 Kalii clavulanas dilutus .......................................................... 3381
Insulini zinci suspensio iniectabilis........................................ 2778 Kalii dihydrogenophosphas .................................................... 3382
Insulinum aspartum ............................................................... 2762 Kalii hydrogenoaspartas hemihydricus................................. 3383
Insulinum biphasicum iniectabile ......................................... 2770 Kalii hydrogenocarbonas ........................................................ 3384
Insulinum bovinum ......................................................... 9.3-4929 Kalii hydrogenotartras............................................................ 3384
Insulinum glarginum ....................................................... 9.5-5699 Kalii hydroxidum ............................................................. 9.4-5430
Insulinum humanum.............................................................. 2768 Kalii iodidum........................................................................... 3385
Insulinum isophanum biphasicum iniectabile ..................... 2770 Kalii metabisulfis..................................................................... 3386
Insulinum isophanum iniectabile .......................................... 2771 Kalii natrii tartras tetrahydricus ........................................... 3388
Insulinum lisprum................................................................... 2771 Kalii nitras ............................................................................... 3386
Insulinum porcinum ........................................................ 9.3-4931 Kalii perchloras........................................................................ 3387
Insulinum solubile iniectabile ................................................ 2771 Kalii permanganas .................................................................. 3388
Interferoni alfa-2 solutio concentrata ................................... 2780 Kalii sorbas .............................................................................. 3389
Interferoni beta-1a solutio concentrata.......................... 9.5-5701 Kalii sulfas................................................................................ 3389
Interferoni gamma-1b solutio concentrata ........................... 2785 Kanamycini monosulfas ......................................................... 2846
int-rac-α-Tocopherolum ......................................................... 3800 Kanamycini sulfas acidus ....................................................... 2845
int-rac-α-Tocopherylis acetas................................................. 3802 Kaolinum ponderosum ........................................................... 2847
Iobenguani (123I) solutio iniectabilis ...................................... 1155 Ketamini hydrochloridum...................................................... 2847
Iobenguani (131I) solutio iniectabilis ad usum Ketoconazolum ........................................................................ 2849
diagnosticum.......................................................................... 1156 Ketoprofenum .......................................................................... 2851
Iobenguani (131I) solutio iniectabilis ad usum Ketorolacum trometamolum.................................................. 2853
therapeuticum........................................................................ 1157 Ketotifeni hydrogenofumaras................................................. 2854
Iobenguani sulfas ad radiopharmaceutica..................... 9.2-4435 Kryptonum (81mKr) ad inhalationem ................................... 1159
Iodinati (125I) humani albumini solutio iniectabilis ............ 1152
Iodixanolum ............................................................................ 2788 L
Iodomethylnorcholesteroli (131I) solutio iniectabilis ............. 1159 Labetaloli hydrochloridum.............................................. 9.4-5397
Iodum ....................................................................................... 2788 Lacca......................................................................................... 3547
Iohexolum ................................................................................ 2791 Lacosamidum ................................................................... 9.5-5709
Iopamidolum ........................................................................... 2795 Lactitolum monohydricum ............................................. 9.3-4939
Iopromidum............................................................................. 2798 Lactosum ........................................................................... 9.3-4940
Iotrolanum............................................................................... 2801 Lactosum monohydricum................................................ 9.3-4942
Ipecacuanhae extractum fluidum normatum ...................... 1395 Lactulosum ....................................................................... 9.5-5710
Ipecacuanhae pulvis normatus .............................................. 1395 Lactulosum liquidum....................................................... 9.5-5712
Ipecacuanhae radix ................................................................. 1397 Lamivudinum .......................................................................... 2871
Ipecacuanhae tinctura normata ............................................ 1398 Lamotriginum ......................................................................... 2873
Ipratropii bromidum............................................................... 2805 Lansoprazolum ................................................................. 9.3-4945
Irbesartanum ........................................................................... 2807 Lanugo cellulosi absorbens ..................................................... 3915
Irinotecani hydrochloridum trihydricum ...................... 9.3-4933 Lanugo gossypii absorbens...................................................... 2172
Isatidis radix ............................................................................ 1399 Lauromacrogolum 400 ........................................................... 2876
Isoconazoli nitras .................................................................... 2811 Lavandulae aetheroleum ................................................. 9.5-5615
Isoconazolum........................................................................... 2810 Lavandulae flos................................................................. 9.5-5614
Isofluranum ............................................................................. 2813 Leflunomidum ......................................................................... 2879
Isoleucinum.............................................................................. 2814 Leonuri cardiacae herba......................................................... 1444
Isomaltum ......................................................................... 9.4-5392 Letrozolum ............................................................................... 2880
Isoniazidum ...................................................................... 9.5-5703 Leucinum ................................................................................. 2881
Isoprenalini hydrochloridum ................................................. 2817 Leuprorelinum ......................................................................... 2883
Isoprenalini sulfas ................................................................... 2818 Levamisoli hydrochloridum ................................................... 2886
Isopropylis isostearas........................................................ 9.2-4563 Levamisolum ad usum veterinarium .................................... 2885
Isopropylis myristas................................................................. 2820 Levetiracetamum.............................................................. 9.4-5399
Isopropylis palmitas ................................................................ 2821 Levistici radix .......................................................................... 1420
Isosorbidi dinitras dilutus....................................................... 2821 Levocabastini hydrochloridum........................................ 9.3-4946
Isosorbidi mononitras dilutus ................................................ 2823 Levocarnitinum ....................................................................... 2890
Isotretinoinum .................................................................. 9.5-5705 Levodopum .............................................................................. 2892
Isoxsuprini hydrochloridum................................................... 2826 Levodropropizinum................................................................. 2893
Isradipinum ............................................................................. 2828 Levomentholum....................................................................... 2894
Itraconazolum ......................................................................... 2829 Levomepromazini hydrochloridum ....................................... 2895
Ivermectinum........................................................................... 2832 Levomepromazini maleas....................................................... 2896
Levomethadoni hydrochloridum ........................................... 2897
J Levonorgestrelum .................................................................... 2899
Josamycini propionas.............................................................. 2839 Levothyroxinum natricum ..................................................... 2902
Josamycinum ........................................................................... 2837 Lichen islandicus ..................................................................... 1392
Juniperi aetheroleum .............................................................. 1404 Lidocaini hydrochloridum...................................................... 2905
Juniperi galbulus ..................................................................... 1404 Lidocainum .............................................................................. 2903
Ligustici chuanxiong rhizoma ......................................... 9.4-5314
K Limonis aetheroleum .............................................................. 1411
Kalii acetas............................................................................... 3375 Lincomycini hydrochloridum................................................. 2906
Kalii bichromas ad praeparationes homoeopathicas ........... 1608 Lini oleum virginale................................................................ 2908
Kalii bromidum ....................................................................... 3376 Lini semen ................................................................................ 1415
Kalii carbonas.......................................................................... 3377 Liothyroninum natricum ....................................................... 2909
Kalii chloridum ....................................................................... 3377 Liquiritiae extractum siccum ad saporandum ..................... 1415
Liquiritiae radix ...................................................................... 1416 Magnoliae biondii flos immaturus ................................. 9.3-4817
Lisinoprilum dihydricum........................................................ 2910 Magnoliae officinalis cortex ............................................ 9.2-4467
Lithii carbonas......................................................................... 2912 Magnoliae officinalis flos ................................................. 9.3-4819
Lithii citras ............................................................................... 2913 Malathionum ........................................................................... 2970
L-Methionini ([11C]methyl) solutio iniectabilis .................... 1161 Maltitolum ............................................................................... 2973
Lobelini hydrochloridum ........................................................ 2913 Maltitolum liquidum .............................................................. 2974
Lomustinum............................................................................. 2914 Maltodextrinum ...................................................................... 2975
Loperamidi hydrochloridum .................................................. 2915 Malvae folium.......................................................................... 1425
Loperamidi oxidum monohydricum ..................................... 2917 Malvae sylvestris flos............................................................... 1424
Lopinavirum ............................................................................ 2918 Mangani gluconas ................................................................... 2976
Loratadinum............................................................................ 2922 Mangani glycerophosphas hydricus....................................... 2976
Lorazepamum.......................................................................... 2924 Mangani sulfas monohydricus ........................................ 9.2-4567
Losartanum kalicum............................................................... 2925 Mannitolum...................................................................... 9.2-4567
Lovastatinum........................................................................... 2927 Maprotilini hydrochloridum .................................................. 2980
Lufenuronum ad usum veterinarium ................................... 2929 Marbofloxacinum ad usum veterinarium ............................ 2981
Lupuli flos ................................................................................ 1388 Marrubii herba ........................................................................ 1557
Lutetii (177Lu) solutio ad radio-signandum ................... 9.3-4799 Masticabilia gummis medicata ....................................... 9.3-4784
Lycii fructus ...................................................................... 9.3-4812 Mastix....................................................................................... 1430
Lycopi herba...................................................................... 9.1-4105 Mate folium ...................................................................... 9.4-5309
Lymecyclinum.......................................................................... 2930 Matricariae aetheroleum ........................................................ 1434
Lynestrenolum ......................................................................... 2932 Matricariae extractum fluidum ............................................. 1433
Lysini acetas............................................................................. 2933 Matricariae flos ....................................................................... 1431
Lysini hydrochloridum............................................................ 2935 Maydis amylum....................................................................... 2969
Lythri herba ............................................................................. 1419 Maydis oleum raffinatum....................................................... 2969
Mebendazolum ........................................................................ 2983
M Meclozini dihydrochloridum........................................... 9.4-5403
Macrogol 20 glyceroli monostearas........................................ 2940 Medroxyprogesteroni acetas ................................................... 2985
Macrogol 40 sorbitoli heptaoleas ........................................... 2941 Mefloquini hydrochloridum ................................................... 2989
Macrogol 6 glyceroli caprylocapras........................................ 2939 Megestroli acetas ..................................................................... 2990
Macrogola ................................................................................ 2950 Megluminum ........................................................................... 2992
Macrogola massae molecularis magnae................................ 2952 Mel ............................................................................................ 2659
Macrogolglyceridorum caprylocaprates.......................... 9.1-4141 Melaleucae aetheroleum......................................................... 1536
Macrogolglyceridorum laurates ...................................... 9.1-4173 Meldonium dihydricum.......................................................... 2993
Macrogolglyceridorum linoleates .................................... 9.1-4174 Meliloti herba .......................................................................... 1437
Macrogolglyceridorum oleates......................................... 9.1-4185 Melissae folii extractum siccum ............................................. 1439
Macrogolglyceridorum stearates ..................................... 9.1-4206 Melissae folium........................................................................ 1438
Macrogolglyceroli cocoates ..................................................... 2948 Meloxicamum.......................................................................... 2994
Macrogolglyceroli hydroxystearas.......................................... 2949 Melphalanum .......................................................................... 2996
Macrogolglyceroli ricinoleas ................................................... 2950 Menadionum ........................................................................... 2998
Macrogoli 15 hydroxystearas ................................................. 2939 Menthae arvensis aetheroleum partim mentholum
Macrogoli 30 dipolyhydroxystearas ....................................... 2941 depletum................................................................................. 1443
Macrogoli aether cetostearylicus ............................................ 2942 Menthae piperitae aetheroleum ............................................. 1478
Macrogoli aether isotridecylicus ............................................ 2943 Menthae piperitae folii extractum siccum ..................... 9.2-4471
Macrogoli aether laurilicus .................................................... 2943 Menthae piperitae folium ................................................ 9.2-4469
Macrogoli aether oleicus ......................................................... 2945 Mentholum racemicum .......................................................... 2998
Macrogoli aether stearylicus................................................... 2947 Menyanthidis trifoliatae folium............................................. 1291
Macrogoli oleas........................................................................ 2944 Mepivacaini hydrochloridum................................................. 2999
Macrogoli stearas..................................................................... 2947 Meprobamatum....................................................................... 3001
Magaldratum ........................................................................... 2953 Mepyramini maleas ................................................................ 3001
Magnesii acetas tetrahydricus ................................................ 2954 Mercaptopurinum ................................................................... 3003
Magnesii aspartas dihydricus.......................................... 9.3-4951 Meropenemum trihydricum................................................... 3004
Magnesii chloridum 4.5-hydricum ........................................ 2957 Mesalazinum ........................................................................... 3005
Magnesii chloridum hexahydricum....................................... 2957 Mesnum............................................................................. 9.2-4569
Magnesii citras......................................................................... 2958 Mesterolonum.......................................................................... 3009
Magnesii citras dodecahydricus ............................................. 2959 Mestranolum............................................................................ 3010
Magnesii citras nonahydricus ................................................ 2959 Metacresolum .......................................................................... 3011
Magnesii gluconas ................................................................... 2960 Metamizolum natricum monohydricum .............................. 3012
Magnesii glycerophosphas....................................................... 2960 Metformini hydrochloridum .................................................. 3014
Magnesii hydrogenophosphas trihydricus ad praeparationes Methadoni hydrochloridum ................................................... 3020
homoeopathicas ..................................................................... 1609 Methanolum ............................................................................ 3022
Magnesii hydroxidum ............................................................. 2961 Methanum ............................................................................... 3021
Magnesii lactas dihydricus ..................................................... 2961 Methanum (2 per centum) in nitrogenio intermixtum.. 9.3-4953
Magnesii oxidum leve ............................................................. 2963 Methenaminum....................................................................... 3023
Magnesii oxidum ponderosum............................................... 2962 Methioninum........................................................................... 3024
Magnesii peroxidum................................................................ 2963 Methotrexatum........................................................................ 3026
Magnesii pidolas...................................................................... 2964 Methylcellulosum .................................................................... 3031
Magnesii stearas ...................................................................... 2965 Methyldopum .......................................................................... 3033
Magnesii subcarbonas levis .................................................... 2956 Methyleni chloridum............................................................... 3034
Magnesii subcarbonas ponderosus......................................... 2955 Methylergometrini maleas...................................................... 3035
Magnesii sulfas heptahydricus ............................................... 2968 Methylhydroxyethylcellulosum .............................................. 3037
Magnesii trisilicas.................................................................... 2968 Methylis nicotinas ................................................................... 3028
Magnesium fluoratum ad praeparationes homoeopathicas.. 1609 Methylis parahydroxybenzoas ............................................... 3029
Methylis parahydroxybenzoas natricus................................. 3591

Methylis salicylas..................................................................... 3030 Naproxenum natricum ........................................................... 3126

Methylphenidati hydrochloridum.......................................... 3037 Nasalia ....................................................................................... 866
Methylphenobarbitalum ......................................................... 3039 Nateglinidum ........................................................................... 3128
Methylprednisoloni acetas............................................... 9.4-5404 Natrii acetas trihydricus ......................................................... 3558
Methylprednisoloni hydrogenosuccinas ................................ 3044 Natrii acetatis ([1-11C]) solutio iniectabilis........................... 1166
Methylprednisolonum............................................................. 3040 Natrii alendronas trihydricus ................................................ 3559
Methylrosanilinii chloridum ........................................... 9.4-5406 Natrii alginas ........................................................................... 3560
Methyltestosteronum............................................................... 3048 Natrii amidotrizoas................................................................. 3561
Methylthioninii chloridum ..................................................... 3049 Natrii aminosalicylas dihydricus ........................................... 3562
Metixeni hydrochloridum....................................................... 3050 Natrii ascorbas......................................................................... 3563
Metoclopramidi hydrochloridum monohydricum ........ 9.4-5409 Natrii aurothiomalas .............................................................. 3565
Metoclopramidum............................................................ 9.4-5407 Natrii benzoas.......................................................................... 3566
Metolazonum........................................................................... 3053 Natrii bromidum ..................................................................... 3567
Metoprololi succinas ............................................................... 3055 Natrii calcii edetas................................................................... 3568
Metoprololi tartras .................................................................. 3056 Natrii calcii pentetas ad radiopharmaceutica ............... 9.3-4800
Metrifonatum .......................................................................... 3058 Natrii caprylas ......................................................................... 3569
Metronidazoli benzoas............................................................ 3060 Natrii carbonas........................................................................ 3570
Metronidazolum...................................................................... 3059 Natrii carbonas decahydricus................................................. 3570
Mexiletini hydrochloridum .................................................... 3062 Natrii carbonas monohydricus .............................................. 3571
Mianserini hydrochloridum ................................................... 3063 Natrii cetylo- et stearylosulfas ......................................... 9.4-5443
Miconazoli nitras .................................................................... 3066 Natrii chloridum ..................................................................... 3573
Miconazolum ........................................................................... 3064 Natrii chromatis (51Cr) solutio sterilis ................................... 1167
Midazolamum ......................................................................... 3069 Natrii citras.............................................................................. 3574
Milbemycinum oximum ad usum veterinarium........... 9.2-4570 Natrii cromoglicas ................................................................... 3574
Millefolii herba ........................................................................ 1564 Natrii cyclamas........................................................................ 3576
Minocyclini hydrochloridum dihydricum...................... 9.2-4573 Natrii dihydrogenophosphas dihydricus ............................... 3577
Minoxidilum ............................................................................ 3072 Natrii docusas.......................................................................... 2309
Mirtazapinum ......................................................................... 3074 Natrii fluoridi (18F) solutio iniectabilis.................................. 1168
Misoprostolum......................................................................... 3075 Natrii fluoridum...................................................................... 3579
Mitomycinum .......................................................................... 3078 Natrii fusidas ........................................................................... 3579
Mitoxantroni hydrochloridum............................................... 3079 Natrii glycerophosphas hydricus ............................................ 3582
Modafinilum............................................................................ 3081 Natrii hyaluronas .................................................................... 3583
Molgramostimi solutio concentrata................................ 9.5-5719 Natrii hydrogenocarbonas ...................................................... 3585
Molsidominum ........................................................................ 3084 Natrii hydroxidum .................................................................. 3585
Mometasoni furoas .......................................................... 9.5-5721 Natrii iodidi (123I) solutioad radio-signandum..................... 1170
Mometasoni furoas monohydricus ................................. 9.5-5724 Natrii iodidi (123I) solutio iniectabilis .................................... 1169
Montelukastum natricum ............................................... 9.1-4179 Natrii iodidi (131I) capsulae ad usum diagnosticum............. 1170
Moranteli hydrogenotartras ad usum veterinarium............ 3090 Natrii iodidi (131I) capsulae ad usum therapeuticum........... 1171
Morphini hydrochloridum ..................................................... 3091 Natrii iodidi (131I) solutio........................................................ 1172
Morphini sulfas ....................................................................... 3093 Natrii iodidi (131I) solutio ad radio-signandum.................... 1173
Moutan cortex .................................................................. 9.4-5311 Natrii iodidum......................................................................... 3586
Moxidectinum ad usum veterinarium .................................. 3095 Natrii iodohippuras dihydricus ad radiopharmaceutica...... 9.2-
Moxifloxacini hydrochloridum .............................................. 3098 4438
Moxonidinum.......................................................................... 3100 Natrii iodohippurati (123I) solutio iniectabilis....................... 1175
Mucores ad producta allergenica........................................... 3094 Natrii iodohippurati (131I) solutio iniectabilis....................... 1176
Mupirocinum........................................................................... 3101 Natrii lactatis solutio .............................................................. 3586
Mupirocinum calcicum........................................................... 3102 Natrii laurilsulfas ............................................................. 9.1-4200
Musci medicati .......................................................................... 859 Natrii lauroylsarcosinas ad usum externum ........................ 3589
Mycophenolas mofetil ............................................................. 3104 Natrii metabisulfis................................................................... 3591
Mycophenolatum natricum.................................................... 3105 Natrii molybdas dihydricus.................................................... 3592
myo-Inositolum ....................................................................... 2761 Natrii molybdatis (99Mo) fissione formati solutio ................ 1176
Myristicae fragrantis aetheroleum......................................... 1455 Natrii nitris .............................................................................. 3593
Myrrha .............................................................................. 9.2-4468 Natrii nitroprussias ................................................................. 3593
Myrrhae tinctura.............................................................. 9.2-4469 Natrii perboras hydricus......................................................... 3594
Myrtilli fructus recens............................................................. 1276 Natrii pertechnetatis (99mTc) acceleratore formati solutio
Myrtilli fructus recentis extractum siccum raffinatum et iniectabilis ....................................................................... 9.3-4801
normatum .............................................................................. 1361 Natrii pertechnetatis (99mTc) fissione formati solutio
Myrtilli fructus siccus.............................................................. 1275 iniectabilis .............................................................................. 1178
Natrii pertechnetatis (99mTc) sine fissione formati solutio
N iniectabilis .............................................................................. 1180
Nabumetonum......................................................................... 3109 Natrii phenylbutyras ............................................................... 3595
N-Acetyltryptophanum........................................................... 1639 Natrii phosphatis (32P) solutio iniectabilis ............................ 1180
N-Acetyltyrosinum .................................................................. 1641 Natrii picosulfas....................................................................... 3596
Nadololum ............................................................................... 3110 Natrii polystyrenesulfonas ...................................................... 3597
Nadroparinum calcicum ................................................. 9.5-5731 Natrii propionas ...................................................................... 3598
Naftidrofuryli hydrogenooxalas............................................. 3113 Natrii pyrophosphas decahydricus ad radiopharmaceutica.. 9.2-
Naloxoni hydrochloridum dihydricum .......................... 9.3-4957 4439
Naltrexoni hydrochloridum ................................................... 3118 Natrii risedronas 2.5-hydricus ............................................... 3495
Nandroloni decanoas .............................................................. 3120 Natrii salicylas ......................................................................... 3601
Naphazolini hydrochloridum................................................. 3122 Natrii selenis ............................................................................ 3601
Naphazolini nitras .................................................................. 3123 Natrii selenis pentahydricus ................................................... 3602
Naproxenum ............................................................................ 3124 Natrii (S)-lactatis solutio ........................................................ 3587
Natrii stearas ........................................................................... 3605
Natrii stearylis fumaras .......................................................... 3606 Olsalazinum natricum............................................................ 3203

Natrii sulfas anhydricus ......................................................... 3607 Omega-3 acidorum esteri ethylici 60..................................... 3205
Natrii sulfas decahydricus ...................................................... 3607 Omega-3 acidorum esteri ethylici 90.............................. 9.2-4583
Natrii sulfis .............................................................................. 3608 Omega-3 acidorum triglycerida............................................. 3209
Natrii sulfis heptahydricus ..................................................... 3608 Omeprazolum .......................................................................... 3211
Natrii tetrachloroauras dihydricus ad praeparationes Omeprazolum magnesicum.................................................... 3213
homoeopathicas .............................................................. 9.5-5623 Omeprazolum natricum ......................................................... 3214
Natrii thiosulfas....................................................................... 3609 Ondansetroni hydrochloridum dihydricum ......................... 3216
Natrii valproas......................................................................... 3609 Ononidis radix......................................................................... 1493
Neohesperidin-dihydrochalconum......................................... 3130 Ophthalmica .............................................................................. 857
Neomycini sulfas ..................................................................... 3132 Opii extractum siccum normatum ........................................ 1459
Neostigmini bromidum........................................................... 3134 Opii pulvis normatus .............................................................. 1460
Neostigmini metilsulfas.................................................... 9.5-5733 Opii tinctura normata ............................................................ 1463
Neroli aetheroleum.................................................................. 1449 Opium crudum ........................................................................ 1461
Netilmicini sulfas.............................................................. 9.2-4579 Orbifloxacinum ad usum veterinarium................................ 3217
Nevirapinum..................................................................... 9.3-4959 Orciprenalini sulfas................................................................. 3219
Nevirapinum hemihydricum.................................................. 3139 Origani herba .......................................................................... 1464
Niaouli typo cineolo aetheroleum.......................................... 1453 Orphenadrini citras ................................................................ 3220
Nicardipini hydrochloridum ........................................... 9.3-4960 Orphenadrini hydrochloridum .............................................. 3222
Nicergolinum .................................................................... 9.4-5415 Orthosiphonis folium .............................................................. 1402
Nicethamidum ......................................................................... 3155 Oryzae amylum ....................................................................... 3487
Niclosamidum.......................................................................... 3142 Oseltamiviri phosphas ..................................................... 9.2-4585
Niclosamidum monohydricum .............................................. 3143 Ouabainum.............................................................................. 3225
Nicorandilum........................................................................... 3144 Oxacillinum natricum monohydricum................................. 3226
Nicotinamidum ....................................................................... 3145 Oxaliplatinum ......................................................................... 3228
Nicotini ditartras dihydricus.................................................. 3147 Oxazepamum .......................................................................... 3231
Nicotini resinas........................................................................ 3148 Oxcarbazepinum ..................................................................... 3232
Nicotinum ................................................................................ 3146 Oxeladini hydrogenocitras ..................................................... 3234
Nifedipinum............................................................................. 3151 Oxfendazolum ad usum veterinarium.................................. 3235
Nifuroxazidum ........................................................................ 3154 Oxitropii bromidum................................................................ 3236
Nilutamidum ........................................................................... 3156 Oxprenololi hydrochloridum ................................................. 3239
Nimesulidum ........................................................................... 3157 Oxybuprocaini hydrochloridum ............................................ 3239
Nimodipinum .......................................................................... 3158 Oxybutynini hydrochloridum ................................................ 3241
Nitrazepamum......................................................................... 3159 Oxycodoni hydrochloridum ................................................... 3242
Nitrendipinum......................................................................... 3160 Oxygenium............................................................................... 3243
Nitrofuralum............................................................................ 3163 Oxygenium (15O) ..................................................................... 1163
Nitrofurantoinum.................................................................... 3164 Oxygenium 93 per centum ..................................................... 3244
Nitrogenii oxidum ................................................................... 3162 Oxymetazolini hydrochloridum............................................. 3245
Nitrogenium............................................................................. 3165 Oxytetracyclini hydrochloridum............................................ 3249
Nitrogenium oxygenio depletum............................................ 3166 Oxytetracyclinum dihydricum ............................................... 3247
Nizatidinum............................................................................. 3167 Oxytocini solutio concentrata ................................................ 3251
N-Methylpyrrolidonum .......................................................... 3046 Oxytocinum ............................................................................. 3250
Nomegestroli acetas................................................................. 3169
Nonoxinolum 9........................................................................ 3170 P
Noradrenalini hydrochloridum ...................................... 9.3-4962 Paclitaxelum ..................................................................... 9.4-5425
Noradrenalini tartras ...................................................... 9.3-4964 Paeoniae radix alba ................................................................ 1472
Norethisteroni acetas .............................................................. 3175 Paeoniae radix rubra .............................................................. 1471
Norethisteronum ..................................................................... 3174 Pancreatis pulvis...................................................................... 3260
Norfloxacinum ................................................................. 9.3-4965 Pancuronii bromidum ............................................................ 3263
Norfluranum............................................................................ 3179 Pantoprazolum natricum sesquihydricum ........................... 3264
Norgestimatum ........................................................................ 3184 Papaverini hydrochloridum ................................................... 3265
Norgestrelum ........................................................................... 3185 Papaveris rhoeados flos........................................................... 1492
Nortriptylini hydrochloridum......................................... 9.3-4967 Paracetamolum ................................................................ 9.4-5429
Noscapini hydrochloridum hydricum ................................... 3188 Paraffinum liquidum ....................................................... 9.5-5737
Noscapinum ............................................................................. 3187 Paraffinum perliquidum.................................................. 9.5-5737
Notoginseng radix ................................................................... 1454 Paraffinum solidum ................................................................ 3268
Nystatinum .............................................................................. 3189 Paraldehydum ......................................................................... 3271
Parenteralia ............................................................................... 871
O Parnaparinum natricum ........................................................ 3272
Octoxinolum 10....................................................................... 3193 Paroxetini hydrochloridum .................................................... 3272
Octyldodecanolum ........................................................... 9.4-5421 Paroxetini hydrochloridum hemihydricum .......................... 3275
Octylis gallas ............................................................................ 3193 Passiflorae herba...................................................................... 1469
Oenotherae oleum raffinatum................................................ 2444 Passiflorae herbae extractum siccum..................................... 1470
Ofloxacinum ............................................................................ 3195 Pefloxacini mesilas dihydricus ............................................... 3277
Olanzapinum........................................................................... 3196 Pelargonii radix ....................................................................... 1470
Oleae folii extractum siccum.................................................. 1458 Pemetrexedum dinatricum heptahydricum.......................... 3279
Oleae folium............................................................................. 1456 Penbutololi sulfas .................................................................... 3281
Olea herbaria ............................................................................ 848 Penicillaminum ....................................................................... 3282
Olibanum indicum.................................................................. 1392 Pentaerythrityli tetranitras dilutus........................................ 3284
Olivae oleum raffinatum ................................................. 9.3-4973 Pentamidini diisetionas .......................................................... 3286
Olivae oleum virginale..................................................... 9.3-4973 Pentazocini hydrochloridum.................................................. 3287
Olmesartanum medoxomilum............................................... 3201 Pentazocini lactas.................................................................... 3288

Pentazocinum .......................................................................... 3287 Plantaginis ovatae semen ....................................................... 1400

Pentobarbitalum ..................................................................... 3289 Plantaginis ovatae seminis tegumentum............................... 1400
Pentobarbitalum natricum..................................................... 3289 Plantarum medicinalium extracta .......................................... 815
Pentoxifyllinum ....................................................................... 3290 Plasma humanum ad separationem ..................................... 2691
Pentoxyverini hydrogenocitras............................................... 3292 Plasma humanum coagmentatum conditumque ad
Pepsini pulvis ........................................................................... 3293 exstinguendum virum.................................................... 9.2-4555
Pergolidi mesilas...................................................................... 3295 Platycodonis radix............................................................ 9.4-5313
Permethrinum 25:75 ............................................................... 3301 Pollines ad producta allergenica ............................................ 3362
Perphenazinum ....................................................................... 3303 Poloxamera .............................................................................. 3363
Persicariae tinctoriae folium.................................................. 1393 Polyacrylatis dispersio 30 per centum ................................... 3365
Pethidini hydrochloridum ...................................................... 3304 Poly(alcohol vinylicus)..................................................... 9.3-4979
Petroleum ad praeparationes homoeopathicas..................... 1612 Polygalae radix ........................................................................ 1515
Pharmaceutica.................................................................. 9.5-5569 Polygoni avicularis herba ....................................................... 1406
Phenazonum ............................................................................ 3305 Polygoni cuspidati rhizoma et radix...................................... 1481
Pheniramini maleas ................................................................ 3306 Polygoni multiflori radix ........................................................ 1356
Phenobarbitalum..................................................................... 3307 Polygoni orientalis fructus............................................... 9.2-4472
Phenobarbitalum natricum.................................................... 3309 Polymyxini B sulfas................................................................. 3366
Phenolphthaleinum ................................................................. 3311 Polyoxypropyleni aether stearylicus ............................... 9.5-5739
Phenolsulfonphthaleinum....................................................... 3311 Polysorbatum 20...................................................................... 3368
Phenolum ................................................................................. 3310 Polysorbatum 40...................................................................... 3369
Phenoxyethanolum ................................................................. 3312 Polysorbatum 60...................................................................... 3370
Phenoxymethylpenicillinum............................................ 9.1-4189 Polysorbatum 80............................................................... 9.2-4591
Phenoxymethylpenicillinum kalicum ............................. 9.1-4190 Poly(vinylis acetas)........................................................... 9.3-4977
Phentolamini mesilas.............................................................. 3317 Poly(vinylis acetas) dispersio 30 per centum ................. 9.3-4978
Phenylalaninum ...................................................................... 3318 Poria ......................................................................................... 1484
Phenylbutazonum ................................................................... 3319 Povidonum........................................................................ 9.2-4592
Phenylephrini hydrochloridum.............................................. 3322 Povidonum iodinatum............................................................ 3394
Phenylephrinum ...................................................................... 3321 Praeadmixta ad alimenta medicata ad usum veterinarium.. 875
Phenylhydrargyri acetas ......................................................... 3324 Praecursores chimici ad radiopharmaceutica......................... 813
Phenylhydrargyri boras .......................................................... 3324 Praeparationes ad irrigationem ............................................... 880
Phenylhydrargyri nitras.......................................................... 3325 Praeparationes buccales................................................... 9.3-4787
Phenylpropanolamini hydrochloridum................................. 3325 Praeparationes celeres ad ptisanam ........................................ 820
Phenytoinum ........................................................................... 3326 Praeparationes homoeopathicae ..................................... 9.1-4117
Phenytoinum natricum........................................................... 3327 Praeparationes insulini iniectabiles....................................... 2776
Phloroglucinolum .................................................................... 3329 Praeparationes intramammariae ad usum veterinarium..... 861
Phloroglucinolum dihydricum ............................................... 3330 Praeparationes intraruminales ....................................... 9.3-4785
Pholcodinum monohydricum ......................................... 9.1-4192 Praeparationes intra-uterinae ad usum veterinarium .......... 862
Phospholipida ex ovo ad iniectabile ............................... 9.2-4533 Praeparationes liquidae ad usum dermicum.......................... 864
Phospholipida ex soia ad iniectabile .............................. 9.4-5445 Praeparationes liquidae peroraliae................................. 9.3-4785
Phthalylsulfathiazolum........................................................... 3334 Praeparationes liquidae veterinariae ad usum dermicum
Physostigmini salicylas ........................................................... 3335 ......................................................................................... 9.3-4792
Phytomenadionum.................................................................. 3336 Praeparationes molles ad usum dermicum............................. 882
Phytosterolum.......................................................................... 3337 Praeparationes molles veterinariae peroraliae ....................... 890
Picotamidum monohydricum ................................................ 3338 Praeparationes pharmaceuticae in vasis cum pressu............. 880
Pilocarpini hydrochloridum................................................... 3339 Pramipexoli dihydrochloridum monohydricum .................. 3394
Pilocarpini nitras..................................................................... 3340 Pravastatinum natricum ........................................................ 3396
Pimobendanum ad usum veterinarium......................... 9.5-5738 Prazepamum............................................................................ 3397
Pimozidum............................................................................... 3343 Praziquantelum....................................................................... 3398
Pindololum .............................................................................. 3344 Prazosini hydrochloridum...................................................... 3400
Pini pumilionis aetheroleum.................................................. 1340 Prednicarbatum....................................................................... 3401
Pini sylvestris aetheroleum ..................................................... 1480 Prednisoloni acetas.................................................................. 3404
Pioglitazoni hydrochloridum ................................................. 3345 Prednisoloni natrii phosphas ................................................. 3407
Piperacillinum ........................................................................ 3347 Prednisoloni pivalas................................................................ 3406
Piperacillinum natricum ........................................................ 3349 Prednisolonum ........................................................................ 3403
Piperazini adipas..................................................................... 3351 Prednisonum..................................................................... 9.3-4980
Piperazini citras....................................................................... 3352 Pregabalinum ................................................................... 9.2-4595
Piperazinum hydricum ........................................................... 3352 Prilocaini hydrochloridum .............................................. 9.4-5432
Piperis fructus.......................................................................... 1474 Prilocainum ...................................................................... 9.4-5431
Piperis longi fructus ................................................................ 1417 Primaquini diphosphas........................................................... 3415
Piracetamum ........................................................................... 3353 Primidonum ..................................................................... 9.3-4982
Pirenzepini dihydrochloridum monohydricum.................... 3354 Primulae radix......................................................................... 1485
Piretanidum............................................................................. 3356 Probenecidum.......................................................................... 3417
Pirfenidonum........................................................................... 3357 Procainamidi hydrochloridum............................................... 3418
Piroxicamum ........................................................................... 3358 Procaini hydrochloridum ....................................................... 3419
Piscis oleum omega-3 acidis abundans ................................. 2474 Prochlorperazini maleas ......................................................... 3420
Pisi amylum ............................................................................. 3277 Producta ab arte ADN recombinandorum............................. 836
Pivampicillinum ...................................................................... 3359 Producta ab fermentatione....................................................... 830
Pivmecillinami hydrochloridum ........................................... 3361 Producta allergenica ................................................................. 811
Plantae ad ptisanam ................................................................. 820 Producta cum possibili transmissione vectorium
Plantae medicinales ......................................................... 9.2-4419 enkephalopathiarum spongiformium animalium................ 832
Plantae medicinales ad praeparationes homoeopathicas .... 1570 Progesteronum......................................................................... 3420
Plantae medicinales praeparatae............................................. 819 Proguanili hydrochloridum............................................. 9.2-4597
Plantaginis lanceolatae folium............................................... 1497 Prolinum .................................................................................. 3424
Promazini hydrochloridum.................................................... 3425 Riboflavinum ........................................................................... 3484

Promethazini hydrochloridum............................................... 3426 Ricini oleum hydrogenatum................................................... 1966
Propacetamoli hydrochloridum ............................................. 3427 Ricini oleum raffinatum ......................................................... 1967
Propafenoni hydrochloridum................................................. 3428 Ricini oleum virginale............................................................. 1968
Propanolum ............................................................................. 3430 Rifabutinum............................................................................. 3488
Propanthelini bromidum........................................................ 3431 Rifampicinum .......................................................................... 3489
Propofolum .............................................................................. 3432 Rifamycinum natricum .......................................................... 3490
Propranololi hydrochloridum ................................................ 3434 Rifaximinum............................................................................ 3492
Propylenglycoli dicaprylocapras............................................. 3438 Rilmenidini dihydrogenophosphas ........................................ 3494
Propylenglycoli dilauras .................................................. 9.2-4598 Risperidonum .......................................................................... 3496
Propylenglycoli monolauras ................................................... 3439 Ritonavirum............................................................................. 3498
Propylenglycoli monopalmitostearas..................................... 3440 Rivastigmini hydrogenotartras .............................................. 3503
Propylenglycolum .................................................................... 3437 Rivastigminum ........................................................................ 3501
Propylis gallas .......................................................................... 3435 Rizatriptani benzoas ............................................................... 3504
Propylis parahydroxybenzoas ................................................ 3436 Rocuronii bromidum .............................................................. 3506
Propylis parahydroxybenzoas natricus ................................. 3599 Ropiniroli hydrochloridum..................................................... 3508
Propylthiouracilum ................................................................. 3441 Ropivacaini hydrochloridum monohydricum ...................... 3509
Propyphenazonum .................................................................. 3442 Rosae pseudo-fructus .............................................................. 1338
Protamini sulfas ...................................................................... 3443 Rosmarini aetheroleum .......................................................... 1500
Prothrombinum multiplex humanum .................................. 2695 Rosmarini folium .................................................................... 1499
Protirelinum ............................................................................ 3444 Rosuvastatinum calcicum....................................................... 3511
Proxyphyllinum ....................................................................... 3445 Roxithromycinum ................................................................... 3513
Prunellae spica......................................................................... 1327 RRR-α-Tocopherolum............................................................. 3801
Pruni africanae cortex ............................................................ 1490 RRR-α-Tocopherylis acetas .................................................... 3804
Pseudoephedrini hydrochloridum ......................................... 3446 RRR-α-Tocopherylis hydrogenosuccinas............................... 3808
Psyllii semen............................................................................. 1486 Rupatadini fumaras ......................................................... 9.3-4987
Puerariae lobatae radix ................................................... 9.3-4816 Rusci rhizoma .......................................................................... 1296
Puerariae thomsonii radix .............................................. 9.3-4820 Rutosidum trihydricum .......................................................... 3516
Pullulanum .............................................................................. 3447
Pulveres ad usum dermicum.................................................... 874 S
Pulveres perorales...................................................................... 874 Sabalis serrulatae extractum .................................................. 1509
Pyranteli embonas................................................................... 3448 Sabalis serrulatae fructus........................................................ 1512
Pyrazinamidum....................................................................... 3449 Sacchari monopalmitas .......................................................... 3665
Pyridostigmini bromidum ...................................................... 3450 Saccharinum ............................................................................ 3521
Pyridoxini hydrochloridum.................................................... 3451 Saccharinum natricum ........................................................... 3522
Pyrimethaminum .................................................................... 3453 Sacchari sphaerae.................................................................... 3670
Pyrrolidonum .......................................................................... 3453 Sacchari stearas ....................................................................... 3666
Saccharum ............................................................................... 3664
Q Saccharum liquidum........................................................ 9.4-5448
Quercus cortex ......................................................................... 1456 Salbutamoli sulfas .................................................................. 3526
Quetiapini fumaras ................................................................. 3457 Salbutamolum ......................................................................... 3523
Quillajae cortex ....................................................................... 1491 Salicis cortex ............................................................................ 1561
Quinaprili hydrochloridum.................................................... 3459 Salicis corticis extractum siccum ........................................... 1562
Salmeteroli xinafoas................................................................ 3529
R Salmonis domestici oleum ...................................................... 3531
Rabeprazolum natricum......................................................... 3469 Salviae lavandulifoliae aetheroleum ..................................... 1524
Rabeprazolum natricum hydricum ....................................... 3470 Salviae miltiorrhizae radix et rhizoma .......................... 9.1-4108
Racecadotrilum ....................................................................... 3472 Salviae officinalis folium ........................................................ 1505
Raclopridi ([11C]methoxy) solutio iniectabilis ...................... 1165 Salviae sclareae aetheroleum ................................................. 1320
Radiopharmaceutica ................................................................. 832 Salviae tinctura ....................................................................... 1507
Raloxifeni hydrochloridum .................................................... 3473 Salviae trilobae folium............................................................ 1506
Raltegraviri compressi...................................................... 9.5-5744 Sambuci flos............................................................................. 1342
Raltegraviri compressi masticabiles ................................ 9.5-5743 Sanguisorbae radix.................................................................. 1509
Raltegravirum kalicum .................................................... 9.4-5437 Saquinaviri mesilas ................................................................. 3533
Ramiprilum.............................................................................. 3475 Schisandrae chinensis fructus.......................................... 9.1-4110
Ranitidini hydrochloridum ............................................. 9.2-4603 Scopolamini butylbromidum .......................................... 9.5-5694
Rapae oleum raffinatum......................................................... 3479 Scopolamini hydrobromidum ................................................ 2735
Ratanhiae radix....................................................................... 1494 Scopolaminum......................................................................... 2733
Ratanhiae tinctura .................................................................. 1495 Scutellariae baicalensis radix ................................................. 1262
Rectalia....................................................................................... 881 Selamectinum ad usum veterinarium................................... 3535
Remifentanili hydrochloridum........................................ 9.2-4604 Selegilini hydrochloridum ...................................................... 3537
Repaglinidum........................................................................... 3479 Selenii disulfidum.................................................................... 3538
Reserpinum .............................................................................. 3481 Selenium ad praeparationes homoeopathicas ............... 9.2-4494
Resorcinolum ........................................................................... 3481 Semecarpus anacardium ad praeparationes
Rhamni purshianae cortex ..................................................... 1307 homoeopathicas ..................................................................... 1591
Rhamni purshianae extractum siccum normatum .............. 1308 Sennae folii extractum siccum normatum ............................ 1517
Rhei radix................................................................................. 1496 Sennae folium .......................................................................... 1516
Rhenii sulfidi colloidalis et technetii (99mTc) solutio Sennae fructus acutifoliae ...................................................... 1518
iniectabilis .............................................................................. 1183 Sennae fructus angustifoliae................................................... 1519
Ribavirinum............................................................................. 3482 Serinum .................................................................................... 3539
Ribis nigri folium..................................................................... 1290 Serpylli herba ........................................................................... 1559
Riboflavini natrii phosphas .................................................... 3485 Sertaconazoli nitras ................................................................ 3540

Sertralini hydrochloridum............................................... 9.4-5441 Sufentanilum ........................................................................... 3668

Serum bovinum ....................................................................... 1863 Sulbactamum natricum.......................................................... 3671
Sesami oleum raffinatum ................................................ 9.1-4197 Sulfacetamidum natricum...................................................... 3673
Sevofluranum.................................................................... 9.5-5749 Sulfadiazinum ......................................................................... 3674
Sildenafili citras................................................................ 9.2-4611 Sulfadimethoxinum.......................................................... 9.4-5450
Silica ad usum dentalem ........................................................ 3550 Sulfadimethoxinum natricum ad usum veterinarium.. 9.4-5451
Silica colloidalis anhydrica..................................................... 3549 Sulfadimidinum....................................................................... 3678
Silica colloidalis hydrica ......................................................... 3550 Sulfadoxinum .......................................................................... 3680
Silica hydrophobica colloidalis............................................... 3551 Sulfafurazolum ........................................................................ 3681
Silybi mariani extractum siccum raffinatum et normatum.. 1440 Sulfaguanidinum..................................................................... 3681
Silybi mariani fructus ............................................................. 1441 Sulfamerazinum ...................................................................... 3682
Simeticonum ............................................................................ 3553 Sulfamethizolum ..................................................................... 3683
Simvastatinum......................................................................... 3554 Sulfamethoxazolum ................................................................ 3684
Sinomenii caulis ...................................................................... 1466 Sulfamethoxypyridazinum ad usum veterinarium.............. 3685
Sitagliptini compressi .............................................................. 3557 Sulfanilamidum....................................................................... 3686
Sitagliptini phosphas monohydricus............................... 9.1-4198 Sulfasalazinum ................................................................. 9.2-4617
Soiae oleum hydrogenatum.................................................... 3630 Sulfathiazolum......................................................................... 3689
Soiae oleum raffinatum ................................................... 9.3-4992 Sulfinpyrazonum ..................................................................... 3689
Solani amylum ........................................................................ 3390 Sulfur ad praeparationes homoeopathicas............................ 1614
Solidaginis herba ..................................................................... 1375 Sulfur ad usum externum....................................................... 3691
Solidaginis virgaureae herba.................................................. 1376 Sulfuris colloidalis et technetii (99mTc) solutio iniectabilis... 1184
Solifenacini succinas ........................................................ 9.3-4991 Sulindacum .............................................................................. 3692
Solutiones ad conservationem partium corporis.................. 3612 Sulpiridum ............................................................................... 3693
Solutiones ad haemocolaturam haemodiacolaturamque .... 2631 Sultamicillini tosilas dihydricus............................................. 3697
Solutiones ad haemodialysem ................................................ 2628 Sultamicillinum ....................................................................... 3694
Solutiones ad peritonealem dialysem .................................... 3299 Sumatriptani succinas ............................................................ 3699
Solutiones anticoagulantes et sanguinem humanum Suxamethonii chloridum ........................................................ 3701
conservantes........................................................................... 1738 Suxibuzonum........................................................................... 3702
Somatostatinum ...................................................................... 3613
Somatropini solutio concentrata..................................... 9.5-5752 T
Somatropini solutio iniectabilis ............................................. 3620 Tacalcitolum monohydricum ................................................. 3707
Somatropinum.................................................................. 9.5-5750 Tacrolimusum monohydricum ....................................... 9.3-4997
Somatropinum iniectabile ............................................... 9.5-5754 Tadalafilum.............................................................................. 3708
Sophorae japonicae flos .......................................................... 1520 Talcum...................................................................................... 3710
Sophorae japonicae flos immaturus ...................................... 1522 Tamoxifeni citras..................................................................... 3712
Sorbitani lauras ................................................................ 9.1-4203 Tamponae medicatae ................................................................ 887
Sorbitani oleas .................................................................. 9.1-4203 Tamsulosini hydrochloridum ................................................. 3714
Sorbitani palmitas............................................................ 9.1-4204 Tanaceti parthenii herba ........................................................ 1355
Sorbitani sesquioleas ........................................................ 9.1-4205 Tanninum ................................................................................ 3716
Sorbitani stearas ............................................................... 9.1-4205 Taraxaci officinalis herba cum radice ................................... 1332
Sorbitani trioleas .............................................................. 9.1-4206 Taraxaci officinalis radix........................................................ 1333
Sorbitolum ............................................................................... 3625 Technetii (99mTc) bicisati solutio iniectabilis.................. 9.2-4440
Sorbitolum liquidum cristallisabile ....................................... 3626 Technetii (99mTc) et etifenini solutio iniectabilis................... 1185
Sorbitolum liquidum non cristallisabile................................ 3627 Technetii (99mTc) exametazimi solutio iniectabilis............... 1187
Sorbitolum liquidum partim deshydricum ........................... 3628 Technetii (99mTc) gluconatis solutio iniectabilis.................... 1188
Sotaloli hydrochloridum ......................................................... 3629 Technetii (99mTc) humani albumini solutio iniectabilis....... 1189
Spectinomycini dihydrochloridum pentahydricum ............. 3631 Technetii (99mTc) macrosalbi suspensio iniectabilis.............. 1190
Spectinomycini sulfas tetrahydricus ad usum Technetii (99mTc) mebrofenini solutio iniectabilis ......... 9.2-4440
veterinarium .......................................................................... 3633 Technetii (99mTc) medronati solutio iniectabilis ................... 1192
Spicae aetheroleum .......................................................... 9.5-5617 Technetii (99mTc) mertiatidi solutio iniectabilis.................... 1193
Spiramycinum .................................................................. 9.2-4612 Technetii (99mTc) microsphaerarum suspensio iniectabilis .. 1194
Spiraprili hydrochloridum monohydricum .......................... 3638 Technetii (99mTc) oxidronati solutio iniectabilis ................... 1195
Spironolactonum ..................................................................... 3639 Technetii (99mTc) pentetatis solutio iniectabilis .................... 1196
Squalanum............................................................................... 3641 Technetii (99mTc) sestamibi solutio iniectabilis .............. 9.2-4441
Stanni colloidalis et technetii (99mTc) solutio iniectabilis..... 1185 Technetii (99mTc) succimeri solutio iniectabilis..................... 1199
Stanni pyrophosphatis et technetii (99mTc) solutio Teicoplaninum......................................................................... 3717
iniectabilis .............................................................................. 1199 Telmisartanum........................................................................ 3718
Stannosi chloridum dihydricum ............................................ 3644 Temazepamum........................................................................ 3720
Stanozololum ........................................................................... 3644 Temozolomidum .............................................................. 9.4-5455
Stavudinum.............................................................................. 3653 Tenoxicamum................................................................... 9.4-5456
Stephaniae tetrandrae radix................................................... 1357 Terazosini hydrochloridum dihydricum ............................... 3724
Stramonii folium .............................................................. 9.2-4473 Terbinafini hydrochloridum .................................................. 3727
Stramonii pulvis normatus.............................................. 9.2-4475 Terbutalini sulfas .................................................................... 3728
Streptokinasi solutio concentrata........................................... 3658 Terconazolum.......................................................................... 3729
Streptomycini sulfas ................................................................ 3660 Terebinthinae aetheroleum ............................................. 9.4-5316
Strontii (89Sr) chloridi solutio iniectabilis ............................. 1181 Terfenadinum.......................................................................... 3730
Strychnos ignatii ad praeparationes homoeopathicas... 9.3-4827 Teriparatidum ......................................................................... 3732
Strychnos nux-vomica ad praeparationes homoeopathicas.. 9.3- Terlipressinum.................................................................. 9.2-4623
4829 tert-Butylamini perindoprilum.............................................. 3296
Styli ............................................................................................. 884 Testosteroni decanoas ............................................................. 3736
Sucralfatum....................................................................... 9.4-5447 Testosteroni enantas ............................................................... 3738
Sucralosum........................................................................ 9.2-4615 Testosteroni isocaproas ........................................................... 3740
Sufentanili citras ..................................................................... 3669
Testosteroni propionas............................................................ 3741 Tributylis acetylcitras.............................................................. 3833

Testosteronum ......................................................................... 3734 Tricalcii phosphas ................................................................... 1929
Tetracaini hydrochloridum .................................................... 3742 Triclabendazolum ad usum veterinarium............................ 3836
Tetracosactidum...................................................................... 3743 Triethylis citras........................................................................ 3837
Tetracyclini hydrochloridum.................................................. 3746 Trifluoperazini hydrochloridum............................................ 3837
Tetracyclinum.......................................................................... 3744 Triflusalum .............................................................................. 3838
Tetra-O-acetylmannosi triflas ad radiopharmaceutica........ 9.2- Triglycerida media .................................................................. 3839
4443 Triglyceroli diisostearas .......................................................... 3841
Tetrazepamum ........................................................................ 3747 Trigonellae foenugraeci semen............................................... 1354
Tetryzolini hydrochloridum ................................................... 3749 Trihexyphenidyli hydrochloridum......................................... 3841
Thallosi (201Tl) chloridi solutio iniectabilis ........................... 1202 Trimebutini maleas.......................................................... 9.3-5005
Theobrominum........................................................................ 3749 Trimetazidini dihydrochloridum........................................... 3843
Theophyllinum ........................................................................ 3750 Trimethadionum ..................................................................... 3845
Theophyllinum et ethylenediaminum ................................... 3752 Trimethoprimum .................................................................... 3846
Theophyllinum et ethylenediaminum hydricum.................. 3754 Trimipramini maleas.............................................................. 3848
Theophyllinum monohydricum ............................................. 3751 Tri-n-butylis phosphas............................................................ 3834
Thiamazolum .......................................................................... 3755 Tritici aestivi oleum raffinatum............................................. 3937
Thiamini hydrochloridum...................................................... 3756 Tritici aestivi oleum virginale ................................................ 3938
Thiamini nitras ....................................................................... 3758 Tritici amylum ........................................................................ 3937
Thiamphenicolum ................................................................... 3760 Trolaminum............................................................................. 3849
Thiocolchicosidum ex ethanolo cristallisatum............... 9.3-4999 Trometamolum........................................................................ 3851
Thiocolchicosidum hydricum .......................................... 9.3-5002 Tropicamidum......................................................................... 3852
Thiomersalum ......................................................................... 3766 Tropisetroni hydrochloridum................................................. 3853
Thiopentalum natricum et natrii carbonas .......................... 3767 Trospii chloridum.................................................................... 3855
Thioridazini hydrochloridum ................................................ 3770 Troxerutinum ................................................................... 9.3-5007
Thioridazinum ........................................................................ 3768 Trypsinum................................................................................ 3857
Threoninum............................................................................. 3771 Tryptophanum......................................................................... 3858
Thymi herba ............................................................................ 1538 Tuberculini aviarii derivatum proteinosum purificatum ... 3862
Thymi typo thymolo aetheroleum ......................................... 1540 Tuberculini bovini derivatum proteinosum purificatum.... 3863
Thymolum................................................................................ 3772 Tuberculini derivatum proteinosum purificatum ad usum
Tiabendazolum ........................................................................ 3773 humanum .............................................................................. 3864
Tiamulini hydrogenofumaras ad usum veterinarium ......... 3777 Tuberculinum pristinum ad usum humanum ..................... 3861
Tiamulinum ad usum veterinarium...................................... 3774 Tylosini phosphas ad usum veterinarium...................... 9.3-5017
Tianeptinum natricum ........................................................... 3779 Tylosini phosphatis solutio ad usum veterinarium....... 9.3-5012
Tiapridi hydrochloridum ........................................................ 3781 Tylosini tartras ad usum veterinarium.......................... 9.3-5021
Tibolonum................................................................................ 3783 Tylosinum ad usum veterinarium.................................. 9.3-5008
Ticarcillinum natricum........................................................... 3784 Tyrosinum................................................................................ 3870
Ticlopidini hydrochloridum ................................................... 3786 Tyrothricinum ......................................................................... 3871
Tigecyclinum ..................................................................... 9.4-5457
Tiliae flos .................................................................................. 1414 U
Tilidini hydrochloridum hemihydricum ............................... 3788 Ubidecarenonum..................................................................... 3875
Timololi maleas ....................................................................... 3789 Uncariae rhynchophyllae ramulus cum uncis ............... 9.3-4821
Tincturae maternae ad praeparationes homoeopathicas .... 1571 Ureum ...................................................................................... 3877
Tinidazolum............................................................................. 3791 Urofollitropinum .............................................................. 9.4-5463
Tinzaparinum natricum ......................................................... 3792 Urokinasum ...................................................................... 9.4-5464
Tioconazolum .......................................................................... 3793 Urtica dioica ad praeparationes homoeopathicas ................ 1614
Tiotropii bromidum monohydricum .............................. 9.3-5004 Urticae folium.......................................................................... 1450
Titanii dioxidum ..................................................................... 3796 Urticae radix............................................................................ 1452
Tizanidini hydrochloridum .................................................... 3797 Uvae ursi folium ...................................................................... 1264
Tobramycinum ........................................................................ 3799
α-Tocopherylis acetatis pulvis ................................................ 3805
V
Tolbutamidum ........................................................................ 3810
Tolnaftatum............................................................................. 3812 Vaccina ad usum humanum ........................................... 9.5-5571
Tolterodini tartras................................................................... 3813 Vaccina ad usum veterinarium....................................... 9.5-5574
Torasemidum........................................................................... 3815 Vaccinum actinobacillosidis inactivatum ad suem.............. 1085
Tormentillae rhizoma ............................................................. 1542 Vaccinum adenovirosidis caninae vivum ............................. 1028
Tormentillae tinctura.............................................................. 1542 Vaccinum adenovirosis caninae inactivatum....................... 1027
Tosylchloramidum natricum ................................................. 3816 Vaccinum anaemiae infectivae pulli vivum .................. 9.5-5598
Toxinum botulinicum A ad iniectabile................................. 1860 Vaccinum anthracis adsorbatum ab colato culturarum ad usum
Toxinum botulinicum B ad iniectabile ................................. 1861 humanum ................................................................................ 893
Tragacantha............................................................................. 1543 Vaccinum anthracis vivum ad usum veterinarium ............. 1003
Tramadoli hydrochloridum.................................................... 3817 Vaccinum aphtharum epizooticarum inactivatum ad
Tramazolini hydrochloridum monohydricum ..................... 3818 ruminantes...................................................................... 9.5-5597
Trandolaprilum....................................................................... 3819 Vaccinum Bordetellae bronchisepticae vivum ad canem .... 1018
Trapidilum............................................................................... 3822 Vaccinum bronchitidis infectivae aviariae inactivatum...... 1007
Trehalosum dihydricum ......................................................... 3823 Vaccinum bronchitidis infectivae aviariae vivum................ 1008
Tretinoinum............................................................................. 3824 Vaccinum brucellosis (Brucella melitensis stirps Rev. 1) vivum
Triacetinum ............................................................................. 3825 ad usum veterinarium .......................................................... 1024
Triamcinoloni acetonidum .................................................... 3827 Vaccinum bursitidis infectivae aviariae inactivatum .......... 1010
Triamcinoloni hexacetonidum .............................................. 3829 Vaccinum bursitidis infectivae aviariae vivum .................... 1011
Triamcinolonum ..................................................................... 3826 Vaccinum calicivirosis felinae inactivatum .......................... 1053
Triamterenum ......................................................................... 3830 Vaccinum calicivirosis felinae vivum .................................... 1054
Tribenosidum .......................................................................... 3832 Vaccinum chlamydiosidis felinae inactivatum ..................... 1055

Vaccinum cholerae aviariae inactivatum ............................. 1063 Vaccinum inactivatum diarrhoeae vituli coronaviro
Vaccinum cholerae perorale inactivatum ............................... 898 illatae...................................................................................... 1025
Vaccinum Clostridii botulini ad usum veterinarium .......... 1035 Vaccinum inactivatum diarrhoeae vituli rotaviro illatae ... 1026
Vaccinum Clostridii chauvoei ad usum veterinarium......... 1036 Vaccinum influenzae equinae inactivatum .......................... 1051
Vaccinum Clostridii novyi B ad usum veterinarium........... 1036 Vaccinum influenzae inactivatum ad suem ......................... 1087
Vaccinum Clostridii perfringentis ad usum veterinarium .. 1038 Vaccinum influenzae inactivatum ex cellulis corticisque
Vaccinum Clostridii septici ad usum veterinarium ............. 1040 antigeniis praeparatum .......................................................... 945
Vaccinum coccidiosidis vivum ad pullum............................. 1042 Vaccinum influenzae inactivatum ex cellulis virisque integris
Vaccinum colibacillosis fetus a partu recentis inactivatum ad praeparatum ............................................................................ 950
ruminantes............................................................................. 1079 Vaccinum influenzae inactivatum ex corticis antigeniis
Vaccinum colibacillosis fetus a partu recentis inactivatum ad praeparatum ............................................................................ 943
suem ....................................................................................... 1077 Vaccinum influenzae inactivatum ex corticis antigeniis
Vaccinum diarrhoeae viralis bovinae inactivatum.............. 1023 praeparatum virosomale ........................................................ 947
Vaccinum diphtheriae adsorbatum ......................................... 924 Vaccinum influenzae inactivatum ex viris integris
Vaccinum diphtheriae, antigeniis minutum, adsorbatum .... 925 praeparatum ............................................................................ 949
Vaccinum diphtheriae et tetani adsorbatum .......................... 899 Vaccinum influenzae inactivatum ex virorum fragmentis
Vaccinum diphtheriae et tetani, antigeni-o(-is) minutum, praeparatum ............................................................................ 942
adsorbatum.............................................................................. 900 Vaccinum influenzae vivum pernasale ................................... 939
Vaccinum diphtheriae, tetani et hepatitidis B (ADNr) Vaccinum laryngotracheitidis infectivae aviariae vivum .... 1014
adsorbatum.............................................................................. 901 Vaccinum leptospirosis bovinae inactivatum ....................... 1019
Vaccinum diphtheriae, tetani et pertussis ex cellulis integris Vaccinum leptospirosis caninae inactivatum ....................... 1030
adsorbatum.............................................................................. 905 Vaccinum leucosis felinae inactivatum ................................. 1058
Vaccinum diphtheriae, tetani et pertussis sine cellulis ex Vaccinum mannheimiae bovinae inactivatum .................... 1071
elementis praeparatum adsorbatum ..................................... 902 Vaccinum mannheimiae inactivatum ad ovem ................... 1072
Vaccinum diphtheriae, tetani et pertussis sine cellulis Vaccinum meningococcale classis C coniugatum ................... 956
ex elementis praeparatum, antigeni-o(-is) minutum, Vaccinum meningococcale polysaccharidicum....................... 958
adsorbatum.............................................................................. 903 Vaccinum morbi Aujeszkyi ad suem inactivatum ............... 1003
Vaccinum diphtheriae, tetani et poliomyelitidis inactivatum, Vaccinum morbi Aujeszkyi vivum ad suem ad usum
antigeni-o(-is) minutum, adsorbatum .................................. 906 parenteralem.......................................................................... 1005
Vaccinum diphtheriae, tetani, pertussis ex cellulis integris et Vaccinum morbi Carrei vivum ad canem............................. 1029
poliomyelitidis inactivatum adsorbatum.............................. 920 Vaccinum morbi Carrei vivum ad mustelidas ..................... 1045
Vaccinum diphtheriae, tetani, pertussis ex cellulis integris, Vaccinum morbi haemorrhagici cuniculi inactivatum........ 1092
poliomyelitidis inactivatum et haemophili stirpis b coniugatum Vaccinum morbillorum, parotitidis et rubellae vivum .......... 952
adsorbatum..................................................................... 9.5-5590 Vaccinum morbillorum, parotitidis, rubellae et varicellae
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex vivum........................................................................................ 953
elementis praeparatum et haemophili stirpis b coniugatum Vaccinum morbillorum vivum................................................. 955
adsorbatum..................................................................... 9.5-5583 Vaccinum morbi Marek vivum .............................................. 1073
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex elementis Vaccinum morbi partus diminutionis MCMLXXVI inactivatum
praeparatum et hepatitidis B (ADNr) adsorbatum ............. 910 ad pullum............................................................................... 1048
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex elementis Vaccinum Mycoplasmatis galliseptici inactivatum.............. 1075
praeparatum et poliomyelitidis inactivatum adsorbatum .. 911 Vaccinum myxomatosidis vivum ad cuniculum .................. 1076
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex elementis Vaccinum panleucopeniae felinae infectivae inactivatum .. 1056
praeparatum et poliomyelitidis inactivatum, antigeni-o(-is) Vaccinum panleucopeniae felinae infectivae vivum ............ 1057
minutum, adsorbatum............................................................ 913 Vaccinum papillomaviri humani (ADNr) .............................. 936
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex Vaccinum parainfluenzae viri canini vivum ........................ 1032
elementis praeparatum, hepatitidis B (ADNr), poliomyelitidis Vaccinum paramyxoviris 3 aviarii inactivatum ad
inactivatum et haemophili stirpis b coniugatum meleagrem.............................................................................. 1016
adsorbatum..................................................................... 9.5-5585 Vaccinum parotitidis vivum..................................................... 959
Vaccinum diphtheriae, tetani, pertussis sine cellulis ex elementis Vaccinum parvovirosis caninae inactivatum ....................... 1033
praeparatum, poliomyelitidis inactivatum et haemophili Vaccinum parvovirosis caninae vivum ................................. 1034
stirpis b coniugatum adsorbatum................................. 9.5-5587 Vaccinum parvovirosis inactivatum ad suem ...................... 1089
Vaccinum encephalitidis ixodibus advectae inactivatum...... 988 Vaccinum pasteurellae inactivatum ad ovem....................... 1084
Vaccinum encephalomyelitidis infectivae aviariae vivum .. 1013 Vaccinum pertussis ex cellulis integris adsorbatum............... 963
Vaccinum erysipelatis suillae inactivatum............................ 1104 Vaccinum pertussis sine cellulis copurificatum adsorbatum.. 962
Vaccinum febris flavae vivum .................................................. 996 Vaccinum pertussis sine cellulis ex elementis praeparatum
Vaccinum febris typhoidis ........................................................ 992 adsorbatum.............................................................................. 960
Vaccinum febris typhoidis polysaccharidicum........................ 990 Vaccinum pestis anatis vivum................................................ 1046
Vaccinum febris typhoidis vivum perorale (stirpis Ty 21a) .. 992 Vaccinum pestis classicae suillae vivum ex cellulis .............. 1104
Vaccinum furunculosidis inactivatum ad salmonidas cum Vaccinum pneumococcale polysaccharidicum........................ 967
adiuvatione oleosa ad iniectionem ............................... 9.2-4427 Vaccinum pneumococcale polysaccharidicum coniugatum
Vaccinum haemophili stirpi b et meningococcale classis C adsorbatum.............................................................................. 965
coniugatum .............................................................................. 926 Vaccinum pneumoniae enzooticae suillae inactivatum ...... 1086
Vaccinum haemophili stirpis b coniugatum .................. 9.5-5592 Vaccinum poliomyelitidis inactivatum ................................... 969
Vaccinum hepatitidis A inactivatum adsorbatum ................. 931 Vaccinum poliomyelitidis perorale ................................. 9.1-4091
Vaccinum hepatitidis A inactivatum adsorbatum et febris Vaccinum pseudopestis aviariae inactivatum ...................... 1080
typhoidis polysaccharidicum.................................................. 929 Vaccinum pseudopestis aviariae vivum ................................ 1082
Vaccinum hepatitidis A inactivatum et hepatitidis B (ADNr) Vaccinum rabiei ex cellulis ad usum humanum .................... 976
adsorbatum.............................................................................. 930 Vaccinum rabiei inactivatum ad usum veterinarium ......... 1093
Vaccinum hepatitidis A inactivatum virosomale ................... 932 Vaccinum rabiei perorale vivum ad vulpem et
Vaccinum hepatitidis B (ADNr) .............................................. 935 nyctereutem ........................................................................... 1096
Vaccinum hepatitidis viralis anatis stirpis I vivum ............. 1047 Vaccinum rhinitidis atrophicantis ingravescentis suillae
Vaccinum herpesviris equini inactivatum ............................ 1050 inactivatum............................................................................ 1090
Vaccinum rhinotracheitidis infectivae bovinae Via praeparandi stirpes homoeopathicas et potentificandi .. 9.2-
inactivatum............................................................................ 1067 4479
Vaccinum rhinotracheitidis infectivae bovinae vivum ........ 1068 Vigabatrinum........................................................................... 3906
Vaccinum rhinotracheitidis infectivae vivum ad Vinblastini sulfas ..................................................................... 3907
meleagrem.............................................................................. 1107 Vincristini sulfas...................................................................... 3908
Vaccinum rhinotracheitidis viralis felinae inactivatum ...... 1059 Vindesini sulfas................................................................. 9.3-5034
Vaccinum rhinotracheitidis viralis felinae vivum ................ 1060 Vinorelbini tartras............................................................ 9.4-5469
Vaccinum rotaviri vivum perorale .......................................... 978 Vinpocetinum .......................................................................... 3914
Vaccinum rubellae vivum......................................................... 981 Violae herba cum flore............................................................ 1558
Vaccinum Salmonellae Enteritidis inactivatum ad Vitamini A synthetici densati pulvis .............................. 9.3-5037
pullum .................................................................................... 1097 Vitaminum A........................................................................... 3917
Vaccinum Salmonellae Enteritidis vivum perorale ad Vitaminum A syntheticum densatum oleosum ............. 9.3-5036
pullum .................................................................................... 1098 Vitaminum A syntheticum, solubilisatum densatum in aqua
Vaccinum Salmonellae Typhimurium inactivatum ad dispergibile ...................................................................... 9.3-5038
pullum .................................................................................... 1100 Voriconazolum ........................................................................ 3921
Vaccinum Salmonellae Typhimurium vivum perorale ad
pullum .................................................................................... 1101 W
Vaccinum tenosynovitidis viralis aviariae vivum ................ 1017 Warfarinum natricum............................................................ 3927
Vaccinum tetani adsorbatum................................................... 987 Warfarinum natricum clathratum........................................ 3928
Vaccinum tetani ad usum veterinarium ............................... 1106
Vaccinum tuberculosis (BCG) cryodesiccatum....................... 895
X
Vaccinum varicellae vivum ...................................................... 994
Vaccinum variolae gallinaceae vivum ................................. 1064 Xanthani gummi .............................................................. 9.2-4627
Vaccinum variolae vivum......................................................... 983 Xenoni (133Xe) solutio iniectabilis .......................................... 1204
Vaccinum vibriosidis aquae frigidae inactivatum ad Xylazini hydrochloridum ad usum veterinarium ................ 3948
salmonidas ...................................................................... 9.2-4428 Xylitolum ................................................................................. 3949
Vaccinum vibriosidis inactivatum ad salmonidas ....... 9.2-4429 Xylometazolini hydrochloridum ............................................ 3951
Vaccinum viri parainfluenzae bovini vivum ........................ 1021 Xylosum.................................................................................... 3953
Vaccinum viri syncytialis meatus spiritus bovini vivum..... 1022
Vaccinum yersiniosidis inactivatum ad salmonidas..... 9.2-4430 Y
Vaccinum zonae vivum ............................................................ 982 Yohimbini hydrochloridum.................................................... 3957
Vaginalia .................................................................................... 887
Valacicloviri hydrochloridum ......................................... 9.3-5029 Z
Valacicloviri hydrochloridum hydricum........................ 9.3-5032 Zanamivirum hydricum......................................................... 3961
Valerianae extractum aquosum siccum ................................ 1549 Zanthoxyli bungeani pericarpium......................................... 1565
Valerianae extractum hydroalcoholicum siccum ................. 1550 Zidovudinum ........................................................................... 3962
Valerianae radix............................................................... 9.1-4111 Zinci acetas dihydricus ........................................................... 3964
Valerianae radix minutata.............................................. 9.1-4113 Zinci acexamas ........................................................................ 3965
Valerianae tinctura ................................................................. 1554 Zinci chloridum ....................................................................... 3967
Valinum.................................................................................... 3890 Zinci gluconas.......................................................................... 3967
Valnemulini hydrochloridum ad usum veterinarium ......... 3892 Zinci oxidum ........................................................................... 3968
Valsartanum ............................................................................ 3895 Zinci stearas ............................................................................. 3968
Vancomycini hydrochloridum................................................ 3896 Zinci sulfas heptahydricus ...................................................... 3969
Vanillinum ............................................................................... 3898 Zinci sulfas hexahydricus ....................................................... 3970
Vardenafili hydrochloridum trihydricum ............................. 3898 Zinci sulfas monohydricus...................................................... 3970
Vaselinum album .................................................................... 3270 Zinci undecylenas.................................................................... 3970
Vaselinum flavum ................................................................... 3271 Zingiberis rhizoma .................................................................. 1367
Vecuronii bromidum ....................................................... 9.1-4211 Ziprasidoni hydrochloridum monohydricum....................... 3971
Vedaprofenum ad usum veterinarium.................................. 3901 Ziprasidoni mesilas trihydricus ............................................. 3973
Venlafaxini hydrochloridum.................................................. 3902 Zolmitriptanum................................................................ 9.5-5759
Verapamili hydrochloridum................................................... 3904 Zolpidemi tartras .................................................................... 3975
Verbasci flos ............................................................................. 1445 Zopiclonum.............................................................................. 3976
Verbenae citriodorae folium .................................................. 1412 Zuclopenthixoli decanoas ....................................................... 3978
Verbenae herba........................................................................ 1555

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EURO PEAN

Published in accordance with the

© Council of Europe, 67075 Strasbourg Cedex, France - 2017

CONTENTS OF SUPPLEMENT 9.5 xlv

GENERAL CHAPTERS 5533

2. Methods of analysis 5533

2.2. Physical and physicochemical methods 5533

2.2.7. Optical rotation 5535

2.4. Limit tests 5537

2.4.20. Determination of elemental impurities 5539

3. Materials for containers and containers 5543

3.2. Containers 5543

4.1.1. Reagents 5549

4.1.3. Buffer solutions 5549

4.2.2. Volumetric solutions 5550

5. General texts 5551

5.4. Residual solvents 5553

5.12. Reference standards 5563

GENERAL MONOGRAPHS 5567

VACCINES FOR HUMAN USE 5581

VACCINES FOR VETERINARY USE 5595

MONOGRAPHS ON SUTURES FOR HUMAN USE 5601

MONOGRAPHS ON SUTURES FOR VETERINARY USE 5607

MONOGRAPHS ON HERBAL DRUGS AND HERBAL DRUG PREPARATIONS 5611

MONOGRAPHS ON HOMOEOPATHIC PREPARATIONS 5619

Note: on the first page of each chapter/section there is a list of contents.

CONTENTS OF SUPPLEMENT 9.5

Isotretinoin (1019) Paraffin, liquid (0239)

MONOGRAPHS Follitropin concentrated solution (2286)

TEXTS WHOSE TITLE HAS CHANGED

Codeine monohydrate (0076) (previously Codeine)

The following texts are deleted as of 1 /anuary 2018.

The following text is deleted as of 1 July 2017.

The following text is deleted as of 1 April 2017.

2.2. Physical and physicochemical

5534 See the information section on general monographs (cover pages)

07/2018:20207 - a temperature control system that indicates the temperature

5536 See the information section on general monographs (cover pages)

2.4. Limit tests

5538 See the information section on general monographs (cover pages)

07/2018:20420 peroxide, at various concentrations, can be used to dissolve

Yes Perform closed- V erify procedure

Perform open- or closed-

Figure 2.4.20.-1. - Elemental impurities decision tree: sample preparation

5540 See the information section on general monographs (cover pages)

Perform the Verify the

Perform analysis using

Verify the Yes

Figure 2.4.20.-2. - Elemental impurities decision tree: measurement

ACCURACY INTERMEDIA TE PRECISION

5542 See the information section on general monographs (cover pages)

5544 See the information section on general monographs (cover pages)

( 1) See ISO 7864, Sterile hypodermic needles for single use.

5546 See the information section on general monographs (cover pages)

5548 See the information section on general monographs (cover pages)

07/2018:40100 mp: 161 ºC to 162 ºC.

07/2018:40202 1 mL of 0.1 M ammonium and cerium sulfate is equivalent to

4.2.2. VOLUMETRIC SOLUTIONS 0.1 M Cerium sulfate. 3001100.

5550 See the information section on general monographs (cover pages)

5.4. Residual solvents

5552 See the information section on general monographs (cover pages)

07/2018:50400 IMPURITIES: GUIDELINES

LIMITING RESIDUAL SOLVENT LEVELS 3. GENERAL PRINCIPLES