Republic of Iraq

Ministry of higher education and scientific researches

University of Babylon
College of pharmacy
Stage One

The research revolves around the Analytical tool : High

Performance Liquid Chromatography ( HPLC )

preparation : Ameer Ali Mohsen

Supervisor : Dr.Essam Fadel
Introduction:

High performance liquid

chromatography (HPLC) is the premier
analytical technique used in many
pharmaceutical
applications including
potency/purity/performance assays,
pharmacokinetics/ bioanalytical
testing, purification, high-
throughput screening (HTS), In Process
Control (IPC) Monitoring and Quality
Control (QC) testing .

pharmaceutical industry is the

major consumer segment of HPLC and
has been the primary driving force for
higher
throughput and performance. This
article provides a brief review of
significant developments in HPLC
impacting
pharmaceutical analysis in the last
decade.

High performance liquid

chromatography (HPLC) is an integral
analytical tool in assessing drug
product stability. HPLC methods should
be able to separate, detect, and quantify
the various drug-related degradants
that can form on storage or
manufacturing, plus detect and
quantify any drug-related impurities
that may be introduced during
synthesis.

HPLC is the dominant separation

technique in modern pharmaceutical
and biomedical analysis because it
results in highly
efficient separations and in most cases
provides high detection sensitivity.
Most of the drugs in multi component
dosage
forms can be analyzed by HPLC method
because of the several advantages like
rapidity, specificity, accuracy, precision
and ease of automation in this method.
HPLC methods development and
validation play important roles in new
discovery,
development, manufacture of
pharmaceutical drugs and various
other studies related to humans and
animals. An analytical
procedure is developed to test a
defined characteristic of the drug
substance or drug product against
established acceptance
criteria for that characteristic.
: HPLC System

Typical HPLC system consists of the

: following main components

: Solvent Reservoirs .1
Storage of sufficient amount of HPLC
solvents for continuous operation of
the system. Could be equipped with an
online
degassing system and special filters to
isolate the solvent from the influence of
. the environment

: Pump .2
This provides the constant and
continuous flow of the mobile phase
through the system; most modern
pumps allow controlled mixing of
different solvents from different
. reservoirs
 : Injector .3
This allows an introduction (injection)
of the analytes mixture into
the stream of the mobile phase before it
enters the column; most modern
injectors are autosamplers, which allow
programmed injections of different
volumes of samples that are withdrawn
from the vials in the
. autosampler tray

: Column .4
This is the heart of HPLC system; it
actually produces a separation of the
analytes in the mixture . A column is
the place where the mobile
phase is in contact with the stationary
phase, forming an interface with
enormous surface. Most of the
chromatography development in recent
years went toward the design of many
different ways to enhance this
. interfacial contact

: Detector .5
This is a device for continuous
registration of specific physical
properties ( sometimes chemical
. properties)

As well as Data recording and

. processing device

? What is the importance of HPLC

The purpose high performance

liquid chromatography (HPLC)
analysis of any drugs is to confirm
the identity of a drug and provide
quantitative results and also to
monitor the progress of the therapy
of a disease(Measuring presented on
the Fig. 1.) is chromatogram
obtained for the plasma of
depressed patients 12 h before oral
.administration of dexamethasone

.Figure.1
 : HPLC has more applications like

HPLC has an expanding rule in drug .1

.discovery

protein identification and .2

.characterization

.Support of protein chemistry .3

Support of Assay development and .4

. screening

Purity Assessment of compound .5

. libraries. And the list goes on and on

: Conclusion
There are huge possibilities for further
HPLC method development in several
segments like creating new materials
for preparing specific and more
efficient stationary phases and related
with that, discovering new
combinations and modifications of
mobile phases.
In scope of signal registration, new,
more efficient hyphenated systems like
HPLC-MS and HPLC-NMR are invented,
as application of 1H NMR and 13C NMR
technique as well, which are more and
more improving day after day, and
whose application will open huge
possibilities and assistance in medical
diagnostics and in tracking the destiny
of healing substances in body liquids.