18 2

PHARMEUROPA 18.
2
CONTENTS
April 2006
New: Pharmeuropa, Pharmeuropa Bio and

Pharmeuropa Scientific Notes Online
Draft Monographs and General Texts

for Comment
201
Publication of Supplements 5.6 and 5.7
178 Adenosine
New: HELPDESK
180 Avian infectious bursal disease vaccine (inactivated)
Annual Report of Activities of the EDQM
181 Cholera vaccine (inactivated, oral)
General Information
193 Clozapine
Alfuzosin hydrochloride
Clopamide
Electronic version of the 5th Edition of the

European Pharmacopoeia
239
239
241
243
245
247
249
Copovidone
252
193
Drop point (2.2.17)
254
CRS: conditions of sale
194
Estradiol benzoate
256
List of CRS adopted at the November 2005 session
199
Eye preparations
258
List of texts adopted at the November 2005 session
200
Flavoxate hydrochloride
260
Updated work programme of the European Pharmacopoeia

(November 2005)
202
Herbal drugs: sampling and sample preparation (2.8.20)
262
Lidocaine hydrochloride
265
Lincomycin hydrochloride monohydrate
267
Macrogol 40 sorbitol heptaoleate
269
List of codes of groups of experts
203
List of Standard Terms: 5th Edition (2004)
204
Standard terms: new and revised terms
204
Elaboration / Revision of a monograph (Procedure 1)
205
Technical Guide: 4th Edition (2005) available
206
Proceedings of conferences of the EDQM
208
Pharmeuropa Scientific Notes
209
Pantoprazole sodium sesquihydrate
276
Pharmeuropa Bio
210
Pholcodine
278
Knowledge database
203
Preparations for nebulisation: characterisation (2.9.44)
280
Promethazine hydrochloride
283
International Conferences
Methylergometrine hydrogen maleate
270
Nifuroxazide
272
Oregano
274
213 Pyridoxine hydrochloride
Training Sessions on the 5th Edition of the European

Pharmacopoeia: Chemicals
285
Rabbit haemorrhagic disease vaccine (inactivated)
287
Riboavin
289
27-28 April 2006, Chicago, USA
214
Sodium alendronate
291
13-14 November 2006, Dublin, Ireland
228
Sodium hyaluronate
293
St. Johns wort dry extract, quantied
295
Streptokinase concentrated solution
298
Impurities Control: Setting Specifications

for Antibiotics and Peptides
21-22 September 2006, Strasbourg, France
219
New Microbiology Chapters of the European

Pharmacopoeia
2-3 October 2006, Strasbourg, France
223
Trimipramine maleate
301
Vaccines for veterinary use
304
Valaciclovir hydrochloride
309
Illustrations of powdered drugs in herbal monographs:
Certification of Suitability of the

Monographs of the Ph. Eur.
231 Eucalyptus leaf
List of certificates
231
Readers Tribune
235 Copovidone
235 Contents of the USP Forum (Vol. 32, No. 1)
Herbal Reference Standards
PHARMEUROPA Vol. 18, No. 2, April 2006
Boldo leaf
International Harmonisation
315
315
317
317
320
177
THE EUROPEAN PHARMACOPOEIA

5th Edition: initial volume 5.0 (2 volumes) + 8 Supplements (5.1 - 5.8)
Supplements 5.6, 5.7, 5.8 published in 2006
The 5th Edition 5.0 (2 volumes) has been available since June 2004 (for prices and ordering
information please consult http://book.pheur.org). It is comprised of texts that were implemented
on 1st January 2005 and a cumulative list of reagents.
Publication of Supplements
The supplements are not cumulative and are to be kept for the duration of the 5th Edition.
Modifications (revisions/corrections) to texts are indicated by a line in the margin.
Supplement 5.1 has been available since September 2004; it is comprised of texts that were
implemented on 1st April 2005.
Supplement 5.2 has been available since December 2004; it is comprised of texts that were
implemented on 1st July 2005.
Supplement 5.3 has been available since June 2005; it is comprised of texts that were
implemented on 1st January 2006.
Supplement 5.4 has been available since September 2005; it is comprised of texts that were
implemented on 1st April 2006 and a cumulative list of reagents.
Supplement 5.5 has been available since December 2005; it is comprised of texts that will be
implemented on 1st July 2006.
Supplement 5.6 will be available in June 2006; it is comprised of texts that will be
implemented on 1st January 2007.
Supplement 5.7 will be available in September 2006; it is comprised of texts that will be
implemented on 1st April 2007 and a cumulative list of reagents.
_____________________________________________________________________________
Online access is available to the database giving names of reagents,
especially chromatographic columns. The address is:
http://www.pheur.org/knowledge.htm
178
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179
The EDQM HELPDESK

for rapid user support
available at www.pheur.org
IF YOU HAVE A QUESTION

OR YOU WOULD LIKE
TO CONTACT THE EDQM
USE THE NEW EDQM HELPDESK
The EDQM has set up a new
HELPDESK to provide user support
online via the EDQM website.
It will replace the general
e-mail addresses that have been
used until now, and will cover
the activities of the EDQM.
WHAT ARE THE ADVANTAGES OF USING

THIS NEW HELPDESK?
Find an answer to your question through a comprehensive and detailed
list of Frequently Asked Questions (FAQs).
Use an electronic form tailored to the subject of your questions.
Use a personal message box for sending your questions via the
HELPDESK.
Receive an acknowledgment of receipt by return e-mail within a few
minutes with a unique reference number for your question.
Check the status of the response to your questions at any time via
your message box.
To contact the EDQM, the new HELPDESK procedure should be used.
The HELPDESK will enable the EDQM to track your questions and
answer them promptly and consistently.
HOW CAN I CONTACT AND/OR SEND

A QUESTION TO THE EDQM?
Click on the HELPDESK button and select the topic that
corresponds to the subject of your question.
If having read through the FAQs you do not nd a response
to your question, click on the link below the replies to the
FAQs.
Enter your account details. If you do not have an account,
you will need to create one before continuing.
Complete the electronic form with details of your question(s)
and click on validate. Please ensure that all the elds are lled in as
this will facilitate the efcient processing of your question.
All questions received will be considered in our continuous
review of the list of FAQs we publish on the EDQM website. To
nd out more about this new service, visit the HELPDESK today:
http://www.pheur.org/site/page_521.php
For more information, visit:

http://www.pheur.org
180
Annual report of activities of the EDQM
European Directorate for the

Quality of Medicines (EDQM)
Annual Report of Activities - 2005
The activities of the European Directorate for the Quality
of Medicines are described in terms of its 3 main areas of
responsibility:
1.
the European Pharmacopoeia, including

publication and communications activities, and
international relations,
2.
the procedure for Certication of Suitability of

monographs of the Pharmacopoeia,
3.
the European network of Ofcial Medicines

Control Laboratories (OMCLs).
In 2005, pharmacopoeia authorities and licensing

authorities all over the world showed growing interest
in the work of the European Pharmacopoeia, the
Certication of Suitability, and the network of Ofcial
Medicines Control Laboratories.
1. THE EUROPEAN PHARMACOPOEIA
PARTIES TO THE CONVENTION AND OBSERVERS
The European Pharmacopoeia convention has been
signed by 35 parties including the EU and the following
countries: Austria, Belgium, Bosnia and Herzegovina,
Bulgaria, Croatia, Cyprus, the Czech Republic, Denmark,
Estonia, Finland, The Former Yugoslav Republic
of Macedonia, France, Germany, Greece, Hungary,
Ireland, Iceland, Italy, Latvia, Lithuania, Luxembourg,
Malta, Norway, the Netherlands, Portugal, Serbia and
Montenegro (formerly Yugoslavia), Romania, the Slovak
Republic, Slovenia, Spain, Sweden, Switzerland, Turkey,
and the United Kingdom.
There are 3 new observer states at the European
Pharmacopoeia Commission: Brazil, Israel and
Madagascar, bringing the total number to 18, namely:
the World Health Organisation (WHO) plus 4 European
countries (Albania, Georgia, Poland, Ukraine) and
13 non-European countries (Algeria, Australia, Brazil,
Canada, China, Israel, Malaysia, Madagascar, Morocco,
Senegal, Syria, Tunisia, United States (FDA)).
GENERAL ACTIVITIES
The European Pharmacopoeia Commission continued
its work on the preparation of the supplements of the
5th Edition, which entered into force on 1 January 2005.
3 supplements (5.3, 5.4 and 5.5) were published in 2005
with implementation on 1 January 2006, 1 April 2006
and 1 July 2006.
At its 3 sessions in March, June and November 2005,
the European Pharmacopoeia Commission adopted
168 monographs (new and revised). The new procedure
(procedure IV) set up for new products based on
collaboration with the manufacturers and national
control laboratories continued to yield encouraging

results: 14 monographs reached at least the public
enquiry stage and half were adopted. The number of
documents produced (new, revised) was stable: 3025
in 2005 (3190 in 2004). The European Pharmacopoeias
biological standardisation work has been extended to
new areas: gene-transfer medicinal products, cell therapy
products and required control methods. A new chapter on
modern microbiological methods was adopted. A revision
of the assay of inactivated poliomyelitis vaccine will
considerably reduce the number of animals used.
A total of 203 days were devoted to meetings in 2005.
This includes the 3 plenary sessions of the Commission
and the corresponding preparatory meetings, the
meetings of the Groups of Experts and those of the
ad hoc Working Parties (76). This total also includes
the participation of members of the Secretariat in
various other meetings: meetings of the Pharmaceutical
Committee (Brussels) on medicines for human and
veterinary use, meetings of the various working parties
of the Committee for Medicinal Products for Human Use
(CHMP) and of the Committee for Veterinary Medicinal
Products (CVMP) of the EMEA (nearly 20 meetings such
as those of the Quality Working Party, Biotech Working
Party, Veterinary Immunological Products Working
Party, Inspectors Working Party and Herbal Medicinal
Products Working Party). In addition, the EMEA/
EDQM meeting of the chairs of these groups (chair
of the CHMP, chair of the European Pharmacopoeia
Commission) with the participation of the heads of the
scientic services of the EMEA/EDQM gave an overview
of the subjects of mutual interest and projects that could
be set up in the future. Members of the Secretariat also
attended meetings of the Pharmacopoeial Discussion
Group (PDG) for International Harmonisation with
Japan and the United States, preparatory meetings of the
Quality Working Party for ICH (Q4B), meetings of VICH
working parties and meetings to organise and take part
in international scientic conferences and congresses.
INTERNATIONAL HARMONISATION WITH THE
PHARMACOPOEIAS OF THE UNITED STATES
AND JAPAN
The Pharmacopoeial Discussion Group [European
Pharmacopoeia (Ph. Eur.), Japanese Pharmacopoeia
(JP) and United States Pharmacopeia (USP)] met in
Brussels (Belgium) on 9-12 May 2005, and in Chicago
(United States) on 7-10 November 2005, coinciding with
the International Conference on Harmonisation (ICH)
meetings, which was useful for exchanging information
on the progress of work. These meetings were set up
to nalise the harmonisation of a number of general
chapters and monographs. The WHO attended as an
observer.
181
Summary of agreements on harmonisation by the PDG

3 new monographs signed off: Calcium disodium
edetate, Dibasic calcium phosphate dihydrate, and
anhydrous dibasic calcium phosphate.
Revision of 6 monographs: Powdered cellulose,
Microcrystalline cellulose, Sodium starch
glycolate, Benzyl alcohol, Anhydrous lactose, and
Methylcellulose.
3 general chapters signed off: Microbial enumeration
methods, Tests for specied micro-organisms, and
Acceptance criteria for pharmaceutical preparations.
Implementation of harmonised texts
To accelerate the inter-regional implementation
of harmonised texts, the JP has introduced a new
procedure for the rapid implementation of harmonised
general chapters related to the Q6A guideline, since
full harmonisation is achieved only when all 3
pharmacopoeias have published and implemented a
monograph/general chapter.
Regulatory acceptance of pharmacopoeial
interchangeability
To facilitate this recognition, the ICH Steering
Committee has set up an Expert Working Group ICH-4B.
As the 3 pharmacopoeias are made legally binding by
the respective legislations, the regulatory authorities
have to set up a common procedure to allow ofcial
recognition of the 3 pharmacopoeias for texts considered
to be harmonised and therefore interchangeable; the
authorities must also deliver a clear message to the
industries in the 3 regions with a common date for it
entering into force.
2 meetings between the PDG and the Q4B group were
held on 10 May and November 9 2005, to discuss the
regulatory acceptance of harmonised monographs and
general chapters, particularly those of relevance to the
ICH Q6A guideline; the document detailing the roles
and responsibilities of the PDG and Q4B EWG was
discussed with further renement necessary; 5 packages
for harmonised general chapters were submitted by the
PDG to the Q4B group: dissolution, extractable volume,
particulate matter in parenterals, residue on ignition/
sulphated ash, and the sterility test.
Examination of the test for residue on ignition/
sulphated ash was completed by the Q4B group,
and tests, analytical procedures, and acceptance
criteria of the 3 pharmacopoeias will be recognised as
interchangeable by the regulatory authorities in the 3
regions once the harmonised text has been published
and implemented in all 3 regions. This revised text was
adopted by the European Pharmacopoeia Commission:
it will be included in supplement 5.6. The package for
dissolution was submitted to the Q4B group in August
2005. The USP has revised its text for extractable
volume and the PDG is awaiting feedback from the
Q4B group. Particulate matter was discussed at the
meeting of 9 November 2005, and the Q4B group
provided a preliminary report outlining a number of
issues remaining to be resolved in order to achieve
regulatory interchangeability. A number of issues
remain to be resolved for the sterility test in order to
achieve regulatory interchangeability. Additionally,
packages for the PDG harmonised texts on disintegration
182
and uniformity of dosage units are in preparation for

submission to the Q4B group.
Relations with industry associations
2 meetings with pharmaceutical industry
associations were held on 10 May and 8 November
2005 to exchange information on progress with the
current work programme and future harmonisation
needs. As important stakeholders, industry
associations were encouraged to play an active role
in the harmonisation process.
Excipients producers: 2 meetings were held with
Tri-PEC (IPEC Americas, IPEC Europe, Japanese
Pharmaceutical Excipients Council) on 12 May and
10 November 2005 to discuss the work programme
on harmonisation of excipient monographs. Current
issues include the policy for functionality-related
characteristics, use of additives and processing
aids in excipients, co-processed excipients, control
of impurities in excipients, and the future of
harmonisation.
STANDARD TERMS
The list of standard terms has been translated into 5
more languages (Estonian, Latvian, Lithuanian, Maltese
and Romanian), so that these terms are now available
in 27 languages, including all of the ofcial languages
of the 10 new EU member states. An electronic
version of the list of standard terms is available on
the EDQM internet site, in a specialised database. The
corresponding paper version has been available since
the beginning of 2005. Closer collaboration with the EU
(EMEA) has been initiated, with members of the EDQM
participating in EMEA meetings and a representative of
the EMEA participating in EDQM meetings.
COMMUNICATIONS AND PUBLIC RELATIONS
The European Pharmacopoeia Commission reinforced
its communications activities by organising the
following international scientic conferences, seminars,
training sessions and visits of the EDQM or specialised
exhibitions for professionals working in the area of quality
control of medicines. Events of media interest were
organised to commemorate the 10th anniversary of the
European network of to convey to the general public
the importance of the activities of the EDQM/European
Pharmacopoeia in ensuring the quality of all medicines
and in the ght against counterfeit products.
Symposia and conferences
Symposium on the quality of homoeopathic products in
the new European legislative framework, February 2005,
Strasbourg, France
A symposium organised by the EDQM was attended by 83
participants from 15 European countries. Presentations
by the European Commission, European regulatory
authorities, manufacturers associations and professional
associations focused on the current legislative situation,
the practice in different countries in application of the
legislation, and future needs for standardisation via the
European Pharmacopoeia.
A number of intractable issues have arisen and one
aim of the symposium was to work towards consensus
on these and to build a relevant work programme for
the future orientation of the European Pharmacopoeia

Working Party. The question of manufacturing methods
and the particular concerns related to raw materials
of human, animal and microbiological origin were
presented and discussed.
A round-table discussion at the end of the symposium
was aimed at compiling recommendations to be made to
the European Pharmacopoeia Commission.
After the plenary symposium, a regulatory debrieng
session drafted a document to be submitted to the
Commission in March 2005, with a view to reviewing the
work programme on homoeopathic preparations.
The need for input from all interested parties was clearly
identied and a strong wish to continue work at the
European Pharmacopoeia was evident.
Symposium on alternatives to whole cell pertussis
vaccine potency assay, 16 March 2005, Geneva,
Switzerland
This symposium was organised by the EDQM, with
the support of the European Centre for the Validation
of Alternative Methods (ECVAM, JRC, European
Commission) and the participation of the WHO (World
Health Organisation).
The aim of the symposium was to determine whether a
replacement test more acceptable in terms of animal
welfare for the current whole cell pertussis vaccine
potency assay could be evaluated within the Biological
Standardisation Programme. 40 participants from 21
countries took part in the meeting. The signicant
steps in whole cell pertussis vaccine development and
control were outlined, as well as an overview of the
work performed to develop humane and science-based
improvements of whole cell pertussis vaccine quality
control methods.
Information day of the European OMCLs and
10th anniversary (1995-2005), 27 May 2005, Rome, Italy
An information day was organised on 27 May in Rome to
celebrate the 10th Anniversary of the European network
of OMCLs. Participants in this event, which was open
to all, were able to learn more about all of the activities
and structures of the network. Representatives of the
licensing authorities, inspection and various industrial
associations (EFPIA, APIC-COFIC, AESGP, EGA,
EVM) took part in the programme. 2 round-tables for
prospective topics were also set up: the rst concerned
the construction of a partnership between the network
of OMCLs and industry to ght more effectively against
counterfeit medicines, and the second concerned
the development of future activities in the network.
Nearly 300 participants from 33 countries attended this
event. For further details, see the section on Biological
Standardisation.
Certication of suitability of monographs of the
European Pharmacopoeia. Implementation of the
5th Edition - New procedures for revision and renewal of
certicates, 27-28 October 2005, Istanbul, Turkey
2 days of exchanges and lively discussions conrmed
that the procedure for the certication of monographs of
the European Pharmacopoeia is a major tool of growing
importance for guaranteeing the quality of constantly
developing world trade. The procedure also plays an
important role in the implementation of the revised

European Directives (2001/83/EEC as amended by 2004/
27/EEC and 2001/82/EEC as amended by 2004/28/EEC).
The objectives of this conference were particularly
important since they involved reviewing the results
obtained since the last conference (4 years earlier),
and sharing points of view and experiences with the
principal users. The programme facilitated dialogue with
users, and indeed 12 workshops sessions (covering the
procedure for renewals and revisions, deciencies in
dossiers, sterile products and inspections, respectively)
and 56 individual consultations (or One-to-One sessions)
were organised, giving each attendee the opportunity
to express and exchange views with European authority
representatives and the assessors who evaluate the
certication dossiers.
Over 180 representatives involved in the quality of
medicines, from 32 countries including Canada, India,
China, South Korea, Israel and the United States,
participated in this international conference organised
by the EDQM. The success of the conference reected
the dynamism of international activities surrounding the
quality of medicines, and the importance of co-operation
between the different partners involved (EDQM,
European Commission, EMEA, national licensing
authorities, inspection and industries).
Training sessions
The EDQM organised 3 training sessions in 2005: in
London (March 2005), Strasbourg (December 2005),
and its rst training session in the United States (April
2005). This latter session was organised at the request
and with the support of the New Jersey Pharmaceutical
Quality Control Association (NJPQCA), and was attended
by more than 100 delegates from the East Coast of the
United States.
Other training sessions organised at the invitation of
the national authorities also took place in Belgrade,
Serbia (150 attendees, April 2005), Hang Zhou, China
(100 attendees, June 2005), Seoul, South Korea (150
attendees, August 2005) and Beijing, China (150
attendees, September 2005).
Ofcial visits to the EDQM
As part of its regular exchanges with its partners, the
EDQM welcomed the following groups to its premises.
Permanent representatives at the Council of Europe
(January and February 2005)
South Korean delegation (July 2005)
The purpose of this visit was to initiate relations between
the EDQM/European Pharmacopoeia and the Korean
authorities. The programme and the presentations aimed
to describe the role and responsibilities of the EDQM
and the European Pharmacopoeia Commission to the
visitors, and to give an overview of European procedures
and standards. The visit also included a tour of the
EDQM laboratories.
Chinese delegation (September 2005)
Exchanges with the Chinese authorities continued
to be developed, and the EDQM received a visit
from a delegation of representatives of the Chinese
Pharmacopoeia and the State Food and Drug
Administration (SFDA). The presentations and
183
discussions mainly concerned the work of the EDQM

and the European Pharmacopoeia, and the role of the
European network of OMCL. In September 2005, the
EDQM also went to Beijing to be received by the heads of
the SFDA, the Chinese Pharmacopoeia and the Chinese
National Institute for the Control of Pharmaceutical and
Biological Products (NICPB).
organisations. It was attended by over 3000 participants

from all over Europe. The EDQM was one of the
partners in creating this new type of event that brings
together students, lecturers, young professionals and
representatives of the pharmaceutical industries. The
main theme of the event was the interaction between
education, research and practical applications.
Russian delegation (September and December 2005)

In September, the EDQM welcomed a delegation of
Russian representatives of industry and of the Ministry
of Health, interested in the activities of the EDQM
and the European Pharmacopoeia. The permanent
representation of Russia at the Council of Europe was
also received in December. Possible rapprochement was
discussed.
CPhI, June 2005, Shanghai, China
Brazilian delegation (November 2005)

Following a request for observer status from Brazil, an
ofcial delegation visited the EDQM in November 2005
in Strasbourg, and was able to participate for the rst
time in a session of the European Pharmacopoeia
Commission.
Association of Southeast Asian Nations (ASEAN)
The EDQM participated in 2 training sessions under a
work programme initiated by the European Committee
for Standardisation (CEN) and commissioned by the EU
to evaluate and train control laboratories for the testing
of pharmaceutical products on the markets of the various
ASEAN countries.
A delegation of national control laboratories of ASEAN
member states participated in the rst training session
organised by the EDQM on its premises (December
2005).
Student visits (March, May and September 2005)
The EDQM welcomed 2 groups of Dutch students in
March and May 2005. The rst group, consisting of 40
pharmacy students at the University of Utrecht, visited
the EDQM as part of a programme of scientic visits
organised to familiarise students with pharmaceutical
science in other European countries. The second group
consisted of 40 students in international relations from
the University of Groningen.
In September 2005, a group of students studying for
a Masters degree from the Institut des Hautes tudes
Europennes in Strasbourg came to learn about the
activities of the Council of Europe that are carried out in
the EDQM.
Exhibitions and professional meetings
These exhibitions and professional meetings provided
an opportunity to meet users of the European
Pharmacopoeia from Europe and Asia. The EDQM
presented the 5th Edition, its publications and its
services to visitors through stands, presentations and
training sessions adapted to the needs of associations or
authorities.
The EDQM participated in the following events.
Pharmaceutical Sciences Fair and Exhibition, 12-17
June 2005, Nice, France
This fair was organised by the European Federation
for Pharmaceutical Sciences (EUFEPS), with several
EUFEPS Member Societies and other proactive scientic
184
The EDQM participated in the Congress of

Pharmaceutical Ingredients (CPhI) exhibition for the
3rd consecutive year. It was attended by nearly 10 000
visitors in 2005. The EDQM stand received many visits
by participants (1200 visitors in 3 days) who were able
to obtain answers to their questions on European
regulations concerning raw materials for pharmaceutical
use, and the publications and services of the EDQM. The
brochures and catalogues were translated into Chinese to
facilitate understanding.
XpoPharm, September 2005, Seoul, South Korea
At the request of the Korean Food and Drug Association
(KFDA) and the Korean Pharmaceutical Trade
Association (KPTA), the EDQM participated in the
XpoPharm 2005 exhibition during its rst ofcial visit to
South Korea.
Indian Pharmaceutical Congress (IPC), December 2005,
Hyderabad, India
The EDQM was invited to participate in this event by
the organisers. The attendees were mainly academics,
but also included representatives of the authorities and
industries.
European Pharmacopoeia exhibition for the general
public
At the time of the 40th anniversary of the European
Pharmacopoeia, it had been decided to organise an
exhibition aimed at members of the general public, who
often nd the concept of medicines to be mysterious.
This exhibition, entitled Find out about the European
Pharmacopoeia and Medicines, consisted of panels that
explained what a pharmacopoeia was, how it guaranteed
the quality of our medicines, and the history of the
European Pharmacopoeia and its work. The exhibition
content is available, in both English and French,
to authorities, universities, institutions etc. and to
anyone who wishes to use it as an educational tool for a
particular event.
During 2005, the exhibition was presented in several
different locations:
Faculty of Pharmacy, Louis Pasteur University,
Illkirch, Strasbourg (19-29 April);
Administrative centre, Place de lEtoile, Strasbourg
(30 April-26 May) and in the main building of the
Council of Europe as part of the Fte de lEurope;
Meeting of the national pharmacopoeias, Danish
Medicines Agency, Copenhagen, Denmark (30-31
May);
Pharmaceutical Sciences Fair and Exhibition, Nice
(12-17 June).
INTERNET SITE - http://www.pheur.org
Online services continued to be developed for users,
with access through the internet site of the EDQM. 2005

saw a number of very helpful new services being made
available.
Online access to electronic versions of the
publication Pharmeuropa and to the List of Standard
Terms.
Direct registration for conferences, symposia and
training sessions.
The Frequently Asked Questions (FAQ) page was
updated for all of the activities of the EDQM. New
questions and answerswere included and user
feedback was used to revise answers.
A new HELPDESK service was set up in June 2005,
to deal with a growing number of requests for
information on technical and scientic aspects of
European Pharmacopoeia texts, and to provide user
support online. This service can be accessed from
the internet site of the EDQM. After identifying
themselves or opening a personal account, users
can submit their questions. They will then receive
an acknowledgment of receipt which will include a
reference number and a link to a personal message
box. This message box will show the status of the
response to the users question by the EDQM and
nally the reply itself. A record of the questions and
the answers to these questions will be kept for one
year by the system. This new system will improve
the traceability of questions sent to the EDQM and
speed up the response time.
The KNOWLEDGE database continues to be the
central repository of technical and regulatory
information for the texts of the European
Pharmacopoeia. The KNOWLEDGE database is now
connected to the 2 other specialised databases for
reference substances and Certication. For a given
substance, users of the KNOWLEDGE database
have direct access to the list of reference substances
or preparations used in the monograph and the
Certicates of Suitability that have been granted.
PROVIDING REFERENCE SUBSTANCES AND
PREPARATIONS
106 new chemical reference substances (or spectra) and
biological reference preparations were adopted during
the year, bringing the number of substances available to
users of the European Pharmacopoeia to 1761. Extensive
collaborative studies were required for 52 of these
substances to determine the content of the substances
used in the assays. In addition, 118 substances were
replaced and the European Pharmacopoeia laboratory
regularly monitored 405 substances and carried out
quality control tests during the production of 506
batches. The number of chemical reference substances
and biological reference preparations distributed to
users continued to climb (from 135 431 vials in 2004 to
152 983 in 2005), and the number of orders increased
from 17 903 to 20 535. From bulk substances selected
by the European Pharmacopoeia Commission for use
as reference substances, the Production Unit of the
EDQM prepared 512 batches (lling 277 404 vials), and 8
batches by lyophilisation (lling 15 627 vials).
PREPARATION AND DISTRIBUTION OF SAMPLES
2773 new samples (compared with 2651 in 2004) were
received by the EDQM this year. The total number of

samples in stock was about 20 000. 403 studies were
carried out by the European Pharmacopoeia laboratory
to compare or check the analytical methods proposed for
new monographs or for revisions of monographs at the
request of the groups of experts of the Commission. The
Production Unit had to prepare 3128 samples for these
laboratory studies to check the quality of the substances
available on the market (multisource substances) or to
check the robustness of draft European monographs. In
addition, 9264 samples were prepared for distribution
to the various experts of the EDQM (for the elaboration
of monographs and the organisation of collaborative
studies, market surveillance studies, biological
standardisation projects, etc.).
BIOLOGICAL STANDARDISATION
The Biological Standardisation Programme (BSP,
Division IV) continued to pursue the following goals in
the area of standardisation of biologicals:
the establishment of European Pharmacopoeia
(working) standards;
the development and validation of new analytical
methods;
the validation of alternative methods in the
framework of the 3R concept (i.e. the Renement,
Reduction and Replacement of animal experiments).
To this end, collaborative studies are performed involving
all interested partners (e.g. OMCLs and manufacturers).
Participation in the collaborative study is not restricted
to members or observers of the Ph. Eur. Commission.
The results of the collaborative studies are published in
Pharmeuropa Bio, which, since 2001, is referenced in
MEDLINE and Index Medicus of the National Library of
Medicine (USA).
Since its start in 1992, 86 BSP projects have been
initiated and 80 BRPs or replacement batches have been
established.
In the year 2005, the following projects were pursued.
In the eld of vaccines for human use
Feasibility study for establishment of common in
vitro potency assay for inactivated poliomyelitis
vaccine (IPV)
Validation of alternatives to Auszyme ELISA kits for
in vitro potency assay of rDNA hepatitis B vaccines
Validation of serological method for potency assay
of acellular pertussis vaccine
Standardisation of assay for residual pertussis toxin
Standardisation of test on molecular size
distribution of haemophilus type b conjugate
vaccine
Standardisation of human inuenza vaccine
serology
Establishment of BRP and validation of methods for
vaccinia immunoglobulin
Validation of NMR methods for quality control of
polysaccharide vaccines
Establishment of replacement batches for hepatitis
A vaccine BRP
In the eld of vaccines for veterinary use
Establishment of mycoplasma reference strains
BRPs
185
Establishment of equine inuenza antiserum BRP

batch 2
In the eld of blood and plasma products

Establishment of BRP for normal human plasma for
assay of SD-plasma and brin sealant kits
Establishment of human coagulation factor VII
concentrate BRP
Establishment of replacement batches for human
normal immunoglobulin BRP
Establishment of immunoglobulin panel for anti-D
antibodies test BRP
Validation of in vitro assay method for tetanus
immunoglobulin
In the eld of biotechnology products
Establishment of somatropin CRS batch 2
Establishment of heparin sodium BRP batch 3
Establishment of an HPLC potency assay for
interferon alfa2
Establishment of low molecular mass heparin for
calibration BRP batch 2
The studies led to the adoption of the following reference
preparations in 2005.
Newcastle disease vaccine (inactivated) BRPs for the
in vitro potency assay
Immunoglobulin panel for anti-D antibodies test
BRP
Vaccinia immunoglobulin BRP
Human immunoglobulin BRP batch 3
Human immunoglobulin (molecular size) BRP
batch 1
The full reports on the concluded collaborative studies
were published or will be published in Pharmeuropa Bio
2005-1 or 2006-1, respectively.
In 2005, signicant progress was made towards the
establishment of mycoplasma reference strains. The
technically demanding production of a sufcient
number of vials of reference strains for Mycoplasma
hyorhinis, M. synoviae, M. orale, M. fermentans and
Acholeplasma laidlawii has been completed. Depending
on the results of nal suitability checks, it is assumed
that the reference preparations will be made available in
mid-2006. It is intended to make them globally available
in the context of both European and international
harmonisation efforts (VICH).
Strong efforts were made to apply the 3R concept to the
eld of quality control of biologicals. The in vitro potency
assay for Newcastle disease vaccine (inactivated) that
had been validated in a collaborative study in 2004 was
presented to the Ph. Eur. Expert Group 15V and led to a
revision of the monograph; the revised monograph was
adopted by the Ph. Eur. Commission in November 2005.
Furthermore, an international symposium was organised
together with the European Centre for Validation of
Alternative Methods (ECVAM) and with the support
of the WHO in Geneva on 16 March 2005, in order to
evaluate possibilities for replacement of the in vivo
potency test for whole cell pertussis vaccine (Kendrick
test). The proceedings of the symposium have been
published and follow-up activities have been initiated. 3
new projects were initiated in 2005 in the context of the
186
3R concept: one projects aims to replace the challenge

assay for tetanus immunoglobulin with an in vitro assay;
the second attempts to better standardise the assay for
residual pertussis toxin and ultimately replace the in
vivo histamine challenge test with an in vitro assay; the
third extends previous projects on the development of
serological assays to replace the in vivo challenge as the
batch potency test for vaccines containing diphtheria
and tetanus components to acelluar pertussis vaccine.
The goal is to enable the potency assay for vaccines
containing all of these components to be carried
out using serum from the same animals. This will
enormously reduce the number of animals needed for
these assays.
As in previous years, co-operation with international
partners continued; projects to establish common
standards were set up whenever possible with the WHO
Expert Committee on Biological Standardisation (ECBS);
an example includes the establishment of a standard for
low molecular mass heparin for calibration. The project
for the establishment of the immunoglobulin panel for
anti-D antibodies test BRP was run together with the
FDA/CBER.
TRANSLATIONS AND PUBLICATIONS
It should be noted that the European Pharmacopoeia
is published in both ofcial languages of the Council
of Europe, namely English and French. The EDQM
therefore has its own specialised translation service. In
2005, 258 texts were translated from English to French
(equivalent to 1246 pages with 300 words per page) and
186 from French to English (equivalent to 907 pages
with 300 words per page).
In the area of publications, the year 2005 issues of
Pharmeuropa comprised a total of 554 pages in French
and 532 pages in English, Pharmeuropa Bio (issues
in English only) comprised 54 pages, Pharmeuropa
Scientic Notes, a new publication in English only,
comprised 66 pages and the 5th Edition of the European
Pharmacopoeia comprised 4642 pages in French and
4370 in English. The 3 supplements for 2005 of the 5th
Edition comprised 1102 pages in French and 1034 in
English.
The 5th edition consists of 1920 monographs, 293 general
texts and 2297 descriptions of reagents, and is published
in both electronic and printed form.
A Certicate of Authenticity for the EDQM publications
was developed to protect against unauthorised copying.
This certicate contains open and hidden security
features. In addition, these certicates contain a unique
EDQM Publication ID, which serves for registration of
the electronic versions and allows users to verify their
genuine EDQM publication using an online registration.
The cumulative electronic edition of the European
Pharmacopoeia is available in 3 different formats: a
CD-ROM version for individual use, an intranet version
for use within company networks, and an online version
accessible through the internet. All 3 electronic editions
are based on browser technology and feature a powerful
search engine, hyperlinks between monographs, general
methods and reagents, and a direct link to the online
database for reference substances and the knowledge
database. The online version uses an improved interface
with some added features, such as displaying the number

of hits in the table of contents, and improved hitlist
presentation. All 3 electronic formats contain printable
Acrobat PDF les of the texts that are identical to those
of the paper version.
Pharmeuropa (including Pharmeuropa Bio and
Pharmeuropa Scientic Notes) is now also available
online. All issues stretching back to Volume 10 (1998)
have been indexed and are available as searchable
Acrobat PDF les. The online access is included for
subscribers to the printed version of Volume 18 (2006).
The necessary access code can be generated using the
information from the Certicate of Authenticity on the
inside-front cover of Pharmeuropa 18.1.
Access to the online version of Standard Terms has
now been included in the subscription to the printed
version, and the access is handled in the same way as for
Pharmeuropa, using the EPID number on the Certicate
of Authenticity on the inside-front cover.
2. CERTIFICATION OF SUITABILITY
OF MONOGRAPHS OF THE EUROPEAN
PHARMACOPOEIA
286 new applications (including 19 for products with
TSE risk) and 465 requests for revision were received,
in addition to the regular updates following the
publication of revised monographs in the supplements
of the European Pharmacopoeia. 780 (new or revised)
certicates were granted after assessment; a further 350
certicates were revised automatically to refer to the 5th
Edition of the European Pharmacopoeia.
In total, over 2700 applications have been received and
1900 certicates have been granted since the procedure
became operational, and these are regularly updated.
Requests for revision or renewal of certicates, which
increased by 50% compared with 2004, are now dealt
with according to a procedure that has been brought into
line with the procedure for marketing authorisation of
medicines. In particular, the deadlines for examination of
dossiers have been shortened according to well-dened
evaluation criteria based on the Community regulations
classication.
The 4th International Conference on Certication
successfully took place at the end of October 2005 (in
Istanbul). Presentations were made by the various
partners (industrial associations, national and European
authorities), and the conference was attended by 180
participants.
The procedure illustrates the exemplary collaboration
between the partners, namely the working parties of
the CHMP, CVMP, and the European Pharmacopoeia
Commission, which, while consulting Industry (EFPIA,
AESGP, CEFIC/APIC, IFHA, EGA, EAPPI, IPEC), worked
together to nd practical solutions to improve quality
assurance without complicating the administrative
procedures for evaluation. The licensing authorities have
clearly expressed their preference for the Certication
Procedure when there is a European Pharmacopoeia
monograph (guideline on Requirements in relation to
active substances, and implementation of directives
2001/82/EC, 2001/83/EC and 2003/63/EC, as amended).
The 3 Cs (Consultation, Co-ordination, Co-operation)
that characterise the procedure are implemented by

a Steering Committee consisting of the Chairs of the
European Pharmacopoeia Commission, the Joint
CHMP/CVMP Quality Working Party, the CHMP Biotech
Working Party, the CVMP Immunological Products
Working Party, the Herbal Medicinal Products Working
Party, the Inspection Working Party, and representatives
of the European Commission, authorities of
non-EU member states of the European Pharmacopoeia,
the EMEA and the EDQM. The Steering Committee met
twice in 2005, thus ensuring that decisions involving
licensing, pharmacopoeia and certication are taken in a
coherent manner.
In addition to the Steering Committee, which is
responsible for decisions on general policy, 2 technical
advisory boards have been set up, one for chemical
substances and the other for TSE risk substances.
They consist of expert rapporteurs who participate
in the evaluation of dossiers. These boards deal with
any technical or scientic questions raised by the
rapporteurs.
3. NETWORK OF OFFICIAL MEDICINES CONTROL
LABORATORIES (OMCLs)
The network was established in 1995 by an initiative of
EDQM in close co-operation with the Commission of
the European Union and is open to all countries that
have signed the European Pharmacopoeia Convention,
as well as to observers at the European Pharmacopoeia
Commission, provided that the criteria of the network
are fullled.
There are two levels of collaboration:
General activities involving all member states of the
network: all the OMCLs are invited to meetings and
are asked to participate in collaborative studies in all
the areas of general interest;
Activities restricted to the European Economic
Area (EEA): a number of activities take place within
the more restrictive regulatory framework for
medicines in the EU, notably those connected to
the Centralised Marketing Authorisation Procedure,
the Mutual Recognition Procedure (MRP), and the
Ofcial Control Authority Batch Release (OCABR) of
blood and plasma derivatives, human vaccines and
veterinary immunobiologicals.
Networking means sharing of know-how within a pool
of experts, work-sharing, and the mutual recognition
of test results based on commonly agreed procedures,
which consequently saves resources and costs in
testing of medicinal products. For that purpose, the
implementation and maintenance of harmonised quality
management systems (QMSs) (based on ISO/IEC 17025)
among the network members and the assessment
of established systems based on commonly agreed
procedures are required. Since 1997, several instruments
have been in place to help OMCLs work towards these
goals, such as training visits (staff members of an OMCL
visit the facilities of another OMCL to learn about quality
assurance (QA) topics or specic analytical techniques),
tutorials (tutors visit an OMCL to coach the personnel
on QA subjects), mutual joint visits (to observe and give
advice on the implementation of a QMS), and mutual
joint audits (to assess the technical competence and
187
the level of compliance of an OMCL). Annual meetings

also contribute towards networking, bringing together
representatives from the whole network to discuss and
exchange viewpoints on common interests, such as the
independent testing of medicines, and to summarise
the years activities and decide on an action plan for the
coming year. These meetings are organised by the EDQM
and hosted by one of the members of the network on a
rotating basis.
In 2005, the QA programme was specially focused on the

OMCLs of the new EU member states that had not yet
been visited, as a pre-assessment in preparation for the
mutual recognition visit by the Canadian authorities.
ACTIVITIES INVOLVING ALL OMCLS OF THE

NETWORK
Training visits are coordinated and supported by

the EDQM Division IV. In 2005 this programme was
fully developed and actively promoted. The following
training subjects were covered in 2005: chromogenic
assay, ELISA, LC-MS, potency assay and purity tests for
certain vaccines, PCR and molecular size distribution of
haemophilus type b vaccines.
Annual meeting of the plenary network
Training courses
The 10 annual meeting of the plenary network was

held on 23-26 May 2005 at the Istituto Superiore di
Sanit in Rome, Italy, and was attended by almost
200 representatives from 31 countries. For the rst
time, a satellite symposium to the annual meeting,
dedicated to the activities of the OMCL Network, was
organised by EDQM (the OMCL Information Day). The
symposium took place at the University La Sapienza,
Rome, on 27 May 2005, and was attended by about 300
participants from industry and public health services
from 33 countries. In plenary lectures and 2 roundtable discussions the broad spectrum of activities of the
network was presented to a large group of stakeholders.
In particular, the activities in the area of counterfeits of
medicinal products were discussed.
Division IV of the EDQM organised 2 training courses

in 2005, open to all scientic and technical staff of the
OMCL Network. The training session on QA general
issues took place at the EDQM on 19-21 April 2005,
with 54 participants from 26 countries. Another,
more technique-oriented training session in the
chemical-pharmaceutical eld took place at the EDQM
on 24-27 October 2005, with 92 participants from 35
countries (5 participants from4 African countries and
2 from Ukraine were sponsored by the WHO). Both
sessions were very positively rated by the participants
and the possibility of exchanging experiences with other
colleagues of the network was highly appreciated.
th
Quality management systems

The quality assurance (QA) programme of the OMCL
Network covers 4 different domains:
educational (tutorials, training courses);
assistance (training visits);
implementation of the quality management systems
(QMS) (mutual joint visits);
surveillance of the QMS (mutual joint audits).
During 2005, the following activities were carried out
within the OMCL Network as part of the QA programme,
coordinated by the EDQM Division IV:
5 mutual joint audits: 2 in OMCLs testing human
and veterinary medicines (chemical), and 3 in
OMCLs testing human medicines (1 biological, 2
chemical + biological);
7 mutual joint visits: 3 in OMCLs testing human
medicines (1 biological, 1 chemical, 1 chemical +
biological), 3 in OMCLs testing veterinary medicines
(1 biological, 2 chemical + biological) and 1 in an
OMCL testing human and veterinary medicines
(chemical + biological);
8 training visits organised for members of 2 OMCLs
(1 EU and 1 non EU);
1 tutorial carried out in an OMCL testing human
medicines (biological).
In total, since the beginning of the QA programme (Dec
1997), 30 mutual joint audits, 44 mutual joint visits,
2 tutorials and 9 training visits have been carried out
within the OMCL Network. These gures show the
strong commitment of the OMCL Network towards
installing a common approach for upgrading their
quality system to a harmonised high standard and to
benet from each others experience.
188
Another technical training session focusing on methods

used in the biological eld was held on 6-9 February
2006 at the EDQM. A training course for auditors
participating in the mutual joint audits/visits scheme is
foreseen later in 2006.
OMCL Network guidelines
The OMCL Network guideline Qualication of
equipment was adopted at the annual meeting in May
2005. It essentially deals with requirements for HPLC
equipment. Further annexes related to other equipment
such as gas chromatography are under development.
Collaboration with the European co-operation for
Accreditation (EA)
Discussions with representatives of the EA concerning
the recognition of the contribution of OMCLs in setting
up QMSs within their domain are progressing. In
particular, the following guidelines/documents were
approved by the EA as recommendation documents to
be used in the context of quality system audits of OMCLs
and are now also available from the EA website:
Validation of analytical procedures;
Scope of accreditation of Ofcial Medicines Control
Laboratories;
Uncertainty of measurement: Policy on the
estimation and application of uncertainty in
analytical measurement (compliance testing).
Prociency Testing Scheme (PTS) Studies
These studies have become a regular programme within
the network. In 2005, six studies were organised in the
physico-chemical eld, with an average participation of
34 national control laboratories, while in the biological
area 4 studies were organised, involving on average 11
national laboratories. A third PTS programme, agreed
with the WHO in 2004, and scheduled for July 2004 to
June 2006, was further developed. In 2005, 3 out of the
5 studies included in the programme were conducted,

1 was nalised and 1 was prepared. On average,
34 governmental control laboratories from Africa, Asia
(South East and Western Pacic), Europe and Central
and South America participated in each study. The
EDQM also participated in a special ASEAN training
programme for the PTS involving ofcial control
laboratories (central and regional) from 9 countries of
South East Asia sponsored by the EU.
the network;
an IT platform to improve communication between
the members of the network.
General studies on market surveillance (MSSs)
Where necessary, the results of these studies support

the revision of the relevant Ph. Eur. monographs and/or
general chapters and methods. This was the case for
the MSS on sub-division of tablets. As an outcome of
the study, the Ph. Eur. Commission agreed to revise the
monograph on tablets in June 2005.
In 1999, the EDQM initiated the development of a

computer programme for the statistical evaluation of
biological dilution assays in accordance with Chapter
5.3 of the Ph. Eur. At that time, most laboratories of the
OMCL network used software developed in-house, so
there was a strong demand for a common programme to
harmonise the presentation of assay data and the analysis
thereof. The lack of availability of suitable commercial
software resulted in the development of CombiStats,
which has been used to the general satisfaction of
the network since 2000. Initially the software was
only available to OMCLs but as of 1 November 2005
non-OMCL laboratories can also obtain a user licence.
Together with the public release of the software an
ofcial website has been launched at www.pheur.org/
combistats on which an online manual, tutorial,
examples and background information can be found. A
free demonstration version can also be downloaded.
Extranet site - OMCLnet
EU/EEA SPECIFIC ACTIVITIES
The extranet site OMCLnet, which was established

in 2003 and fundamentally restructured in 2004, has
meanwhile become a standard information platform for
OMCLs within the network. It hosts information about
adopted general OMCL guidelines, as well as minutes and
presentations of meetings covering the general activities
of the network as well as the MSS, PTS and QA topics.
At the beginning of 2005, the OMCLs were asked to reregister to obtain new access to the extranet site. The
password will be changed regularly to guarantee data
security. Due to the general acceptance of this IT tool by
the OMCLs, it was decided to make meeting documents
only available on OMCLnet prior to the corresponding
meeting, and not to re-distribute them during the
session. This new policy has proven to work satisfactorily
and contributes to reducing the paper document and
postal distribution costs.
Ofcial Control Authority Batch Release (OCABR) of

biologicals
These studies, aimed at screening the quality of products

commercialised in countries of the OMCL Network,
were carried out for the following preparations,
with the participation of 15 national laboratories on
average: equisetum stem, cadmium in herbal drugs and
trimethoprim (raw material and tablets).
2 additional MSSs on sub-division of tablets and on
uniformity of dosage units (Ph. Eur. general method
2.9.40), each involving 20 or more OMCLs of the
network, were nalised during 2005.
OMCL inventory database project

At the end of 2004/beginning of 2005, a campaign
of questionnaires was initiated by Division IV of the
EDQM, addressed to the OMCLs of the general network
(launched in 4 steps). The purpose of these activities was
to gain information about the competences and knowhow in the eld of quality control of medicinal products
throughout the network. The data collected by the
questionnaires will be the basis for an OMCL inventory
database, scheduled for development in 2006. A list of
specications for this database has been established
with the help of an external IT expert and under the
participation of a representative of the OMCLs in the
4th quarter of 2005. The future database should provide:
basic information on OMCLs of the network (name,
address, contact points, etc.);
basic information on competences available within
Data will be stored in a secure, access-controlled

environment. In a future step it might be envisaged
to grant access to other stakeholders (e.g. licensing
authorities, inspectorates etc.).
CombiStats
For the second consecutive year all 25 EU member

states, including those integrated as of 1 May 2004,
took part in the traditional condential exchange of
information on issues related specically to OCABR
during the annual meeting in specic dedicated sessions.
EEA member states and Switzerland, an MRA partner,
also participated as usual.
In addition, continuing the effort to facilitate the
integration of the new member states into the
harmonised system and following the successful
workshop held for the blood network at the Austrian
Federal Institute for Medicines (BiFA) in Vienna on
1-3 December 2004, a similar training workshop was
held for the vaccine network at the Agence Franaise
de Scurit Sanitaire des Produits de Sant (AFSSAPS)
in Lyon on 4-6 April 2005. 13 representatives from 8
of the 10 new member states and 3 participants from
the candidate states of Bulgaria, Croatia and Romania
participated in discussions, and saw presentations from
representatives of several of the long-standing member
states with a focus on the practical application of batchrelease techniques, including laboratory sessions. The
meeting was concluded with a visit to the Sano-Pasteur
manufacturing and control laboratories sites in Marcy
lEtoile. All participants expressed enthusiasm for this
successful exchange of experiences.
Another important workshop was held on 16-17
November 2005 at the National Institute for Biological
Standards and Control (UK) on the batch release of
oral poliomyelitis vaccines. 10 participants from 5
member states and observers from the EDQM and the
WHO participated in this hands-on workshop to gain
experience in the new transgenic mouse neurovirulence
189
assay, which will replace the neurovirulence assay

in monkeys, and to develop guidelines for the
implementation of training and testing procedures at
OMCLs.
Human biologicals
At the annual meeting, a review of OMCL batch release
activities since 2004 for both blood and vaccines and
special scientic presentations were highlighted. Of key
importance was the revision of the terms of reference
for the network to better integrate the new members
states and to increase the size of the advisory group
to accommodate the new depth of activity. The use
of the established communication network was also
emphasised. An increased interest in work-sharing and
possible subcontract testing between OMCLs for OCABR
was highlighted and will be pursued in 2006.
A revision of the administrative procedure for batch
release of biologicals to fully commit OMCLs to external
assessment of QA standards based on ISO 17025 was
adopted. A revision to the terms of reference for the
network was adopted and a position paper on a common
policy for access to information in the OCABR network
was approved and will be followed up. In a related
issue, an administrative procedure for application of
batch release in the context of Article 58 of EU Council
Regulation 726/2004 was presented.
Blood products and plasma derivatives
The following guideline was newly adopted:
Ofcial control authority protocol for approval of
plasma pools.
The following guideline revisions were adopted:
revisions to all blood and plasma-derived productrelated guidelines and the annex IId of the
administrative procedure for batch release of
biologicals to accommodate the change to plasma
pool approval in the newly adopted guideline (see
above);
revisions to all blood and plasma-derived productrelated guidelines to delete the need for hepatitis
C serology test for plasma pools to be in line with
the relevant Ph. Eur. Monograph, which had been
revised;
revisions to the guideline for clotting factor
concentrates, plasma inhibitor concentrates and
brin sealants to adapt to new products available on
the market;
revisions to the guideline for normal
immunoglobulin to be in line with monograph
revisions.
Human Vaccines
The following new internal procedures/position papers
were adopted in 2005:
to OCABR network members on OCABRnet, a restrictedaccess extranet site maintained by the EDQM.
5 new guidelines were adopted in 2005 as follows:
Approval of poliomyelitis vaccine (oral) (OPV)
monovalent bulk;
Poliomyelitis vaccine (oral) (OPV) trivalent
vaccine;
Hepatitis A (inactivated) and typhoid polysaccharide
combined vaccine (adsorbed) for mix at use format
products;
Pandemic inuenza vaccine;
Varicella vaccine.
The following guideline was approved for external
consultation:
Inuenza vaccine (surface antigen, inactivated,
virosome).
All adopted guidelines and administrative procedures are
available in a booklet published by the EDQM at the end
of December 2005. They are also available for download
on the EDQM website
(http://www.pheur.org/site/page_611.php).
Immunological Veterinary Medicinal Products (IVMPs)
Competent Authorities (CA) and OMCLs involved in
the control of veterinary immunological products took
part in the annual meeting again in 2005 with the
other branches of the OMCL network. The veterinary
participants focused on annual reports of activities and
scientic issues with a brief update on the progress
made on formalising procedures for harmonisation and
transparency of an OCABR system in the EU/EEA as
implemented under the current legislation.
In addition to the annual meeting, over the course of
2005, 2 meetings open to all EU/EEA interested parties
and a conference call were organised by the EDQM.
These and a number of other meetings hosted by the
EU Commission and the Veterinary Pharmaceutical
Committee were held to further advance the nalisation
of procedures and guidelines required for application of
the new Directive for Veterinary Medicine, which came
into force on 31 October 2005.
As a result, the following procedures and guidelines have
been developed and approved by the CA/OMCL network
and Veterinary Pharmaceutical Committee for use during
a pilot phase of 1 year. At the end of the pilot phase
an evaluation including an impact assessment will be
completed in cooperation with industry representatives,
and the future needs of the program determined.
Administrative procedure for harmonised application
of Article 81 of Directive 2001/82/EC as amended by
2004/28/EC (Ofcial batch protocol review OBPR)
Position paper on Japanese encephalitis vaccines

marketed in the EU;
Administrative procedure for application of Article

82 of Directive 2001/82/EC as amended by 2004/
28/EC (Ofcial control authority batch release
OCABR)
Position paper on the need for interaction between

OMCLs and licensing bodies.
Models for submission of batch protocols by a

Marketing Autorisation Holder for OCABR/ OBPR
OCABR procedure for pandemic situations;
Adopted internal procedures/position papers are available
190
Inactivated bacterial vaccines
Live bacterial vaccines

Inactivated viral vaccines
Live viral vaccines
Tuberculin PPD/ brucellin preparations
Product-specic technical guidelines for OCABR
Aujeszkys disease vaccine (inactivated)
Aujeszkys disease vaccine (live)
Brucellosis vaccine (live)
Brucellin preparations
Equine inuenza vaccine (inactivated)
Infectious bovine rhinotracheitis vaccine
(inactivated)
Infectious bovine rhinotracheitis vaccine (live)
Newcastle disease vaccine (inactivated)
Newcastle disease vaccine (live)
Rabies vaccine (inactivated) for veterinary use
Rabies vaccine for foxes (live)
Swine erysipelas vaccine (inactivated)
Swine erysipelas vaccine (live)
Tuberculin PPD, avian
Tuberculin PPD, bovine
All of the above noted procedures and guidelines are
available on the EDQM website for download
(http://www.pheur.org/site/page_634.php).
Meeting with manufacturers associations
Each of the sub-networks for OCABR of biologicals
held separate meetings in Strasbourg with the relevant
manufacturers association in the course of 2005 to allow
for exchange and feedback and to ensure transparency
and good will.
Representatives from the vaccine network met with
members of vaccine manufacturers (the European
Vaccine Manufacturers (EVM) Association and other
independent manufacturers) on 6 October 2005
and representatives from the blood network met
with representatives from the International Plasma
Fractionators Association (IPFA) and the Plasma Protein
Therapeutics Association (PPTA) on 25 November
2005. Representatives from the veterinary network for
IVMPs met with representatives from the International
Federation for Animal Health (IFAH) Europe on 12
December 2005.
Market surveillance for products with a centralised
marketing authorisation
The programme for sampling and testing of centrally
authorised products (CAP) was successfully continued in
2005. Since its implementation in 1999, the programme
has been continuously improved thanks to the
collaboration between all stakeholders:
the European Medicines Agency (EMEA), which is
the sponsor and has the overall responsibility for the
programme;
the EDQM, which coordinates sampling and testing
operations on the basis of the information provided

by the Marketing Authorisation (MA) Holders upon
request from the EMEA and reports to the EMEA the
outcome of sampling and testing operations with
proposals for follow-up actions where necessary
the national inspection services, which perform
product sampling on the market;
the OMCLs of the EU/EEA OMCL Network, which
perform analytical testing of products.
The CAP programme covers medicinal products for
both human and veterinary use. In 2005 it included
39 products (13 biotech products and 26 chemical/
pharmaceutical products). In addition, testing was
also carried out on the active substances of 4 of these
products. These gures correspond to a decrease of
5% compared to the 2004 CAP programme and an
increase of 5% compared to the 2003 CAP programme,
and follow the trend seen with the number of MAs
granted. Products to be included in the 2005 programme
were selected by EMEA expert committees from those
authorised in 2002 (year n-3), thus guaranteeing that
selected products have been launched and manufactured
on a large scale. Some products tested in previous years
were included as well for re-testing.
Market samples were collected for each product in 3
EU/EEA countries in order to have an overview of the
actual product quality of distributed batches. Sampling
took place mainly at wholesalers (75%), with less than
10% in retail or hospital pharmacies, mainly because of
the distribution scheme of centrally authorised products,
which are generally difcult to obtain in sufcient
quantities at the very end (pharmacies/hospital) of the
distribution chain without depleting the essential stock
needed for patient care. Overall, 93 sampling operations
were carried out by national inspection services in
24 EU/EEA countries. The market samples and noncommercially available standards and specic reagents
provided by the manufacturers represented 309 stock
items. Initial storage, coding and dispatching to OMCLs
for testing was dealt with by the EDQM.
Testing was usually performed by 2 OMCLs,
independently from the product manufacturers. A
new testing scheme was successfully introduced for
chemical products. 8 products (about one third) were
tested by a single OMCL. A second OMCL could be
involved at a second stage only in case of analytical
or compliance issues. This was the case for only one
product during 2005. It is intended to gradually phase
in such a testing format so that by 2007 all chemical
products will be tested according to the new scheme.
Given this background, it is planned that the OMCLs
that have successfully undergone external assessment of
their QA system, fullling the requirements for market
surveillance of medicinal products, will progressively be
given preference for participation in such CAP projects.
Overall, 31 OMCLs (human and veterinary sides) from
24 EU/EEA countries took part in the testing phase of
the 2005 programme, and a total of 77 testing operations
were carried out.
In 2005, all tested products were of appropriate quality.
Nevertheless, some minor issues related to the quality
of the analytical documentation (MA dossier and/or
Standard Operating Procedures for quality control) were
191
reported. Difculties were also encountered during the

setting-up of methods, e.g. for automated methods or
unusual techniques. All issues were thus reported to
the EMEA and their scientic experts, when necessary
proposing follow-up actions on the registration dossier
and/or on analytical testing methods. The MA holder
obtains access to the reports via the EMEA.
The collaboration between all parties was facilitated
by the extensive use of IT tools (EMEA Eudralink,
EDQM CAPnet server) and 3 productive meetings at the
EDQM; 3 QA documents were nalised by the Advisory
Group of the CAP programme and adopted during
the annual meeting, which took place in December
2005 in Strasbourg. Participants emphasised that the
implementation of a risk-based approach in the selection
of products to be tested to optimise the use of OMCL
resources should be carefully evaluated.
Post-marketing surveillance of products with an MA
according to the Mutual Recognition Procedure (MRP)
2005 has been an eventful year for the MRP-product
testing programme mainly due to 2 important steps
taken to enhance the programme.
In May 2005, at the annual OMCL meeting, the plenum
decided to continue with a regular programme after
having run a successful trial phase over approximately
4 years. At the same time, several OMCLs, mainly from
the new EU member states, announced their interest
in contributing to the programme in the future.
Meanwhile, Division IV of the EDQM has granted 36
OMCL contacts access to its extranet site MRPnet, which
192
is the shared IT communication platform of the MRPproduct testing group and the EDQM. At the beginning
of 2005, it was agreed henceforth to make all individual
Summary Reporting Sheets available to the MRPnet
users. These reports include concise information about
the outcome of testing of MRP-products performed
in the course of the annual programmes, and provide
complementary information to the results sheets, which
have been established previously on MRPnet.
The year 2005 was also the starting point for the
development of an MRP-product testing database,
which will be a common database hosting MRP-product
information and data on tests performed in the course of
this programme.
4. NEW EDQM BUILDING
The foundation-stone-laying ceremony took place
in Strasbourg (alle Kastner) on 28 April 2005. The
ceremony was opened by Mr. Terry Davis, SecretaryGeneral of the Council of Europe. Speeches were made
by Mr. Robert Grossmann, Deputy Mayor and President
of the Communaut Urbaine de Strasbourg, Ms Fabienne
Keller, Mayor of Strasbourg, Mr Ren van der Linden,
President of the Parliamentary Assembly, and Mr Adam
Daniel Rotfeld, Chairman-in-Ofce of the Committee
of Ministers of the Council of Europe and Minister for
Foreign Affairs of Poland. Work progressed as planned
in 2005 and the building should be ready, barring
unforeseen circumstances, by the end of December 2006.
This new building is critical to the future development of
the EDQM/European Pharmacopoeia and will help it to
respond to new public health needs in Europe.
General Information
General Information
5th EDITION OF THE EUROPEAN PHARMACOPOEIA
ELECTRONIC VERSION
1920 New and Revised Monographs and 293 General Texts
With the electronic version of the 5th Edition of the European Pharmacopoeia you can view 1920 monographs,
293 general texts (including general monographs and methods of analysis), 2297 reagents, and also have a direct
internet link to the most recent catalogue of reference substances, which contains 1755 references.
The electronic format has the following convenient features:
hierarchical table of contents, subject index and keyword search;

hyperlinks in the text of a monograph giving access to information on general methods, reagents and reference
substances used in the monograph;
changed (inserted and deleted) texts indicated in both the HTML version and the Acrobat version;
direct access from a monograph or a general method to the CRS database on the internet;
use of a standard internet browser to access the data;
direct printing of an Acrobat version for each individual monograph;
internet and intranet versions available;
NEW: direct links to the general notices and the list of general monographs from each text.
The electronic version is available in English, in French and in a bilingual version.
PRICES: SEE THE CATALOGUE ON OUR WEBSITE http://book.pheur.org

Our prices are indicated in EUR but we accept payments in national currencies.
________________________________________________________________________________
FREQUENTLY ASKED QUESTIONS:

WHAT IS THE ROLE OF
THE EUROPEAN PHARMACOPOEIA?
The European Pharmacopoeia is the original source of harmonised quality standards for medicines; all of its
published texts have undergone European harmonisation. These texts are mandatory in 34 European countries* and
in the European Union and replace any pre-existing national texts on the same subject.
As laid down in their national legislation, certain member states** may continue to issue a national pharmacopoeia
that republishes all or some of the harmonised European texts (most often General Chapters), translated if necessary
into the national language. In all cases it is the European text that is implemented and made legally binding in all of
the member states.
*Austria, Belgium, Bosnia and Herzegovina, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Portugal, Romania, Serbia and Montenegro, Slovak
Republic, Slovenia, Spain, Sweden, Switzerland, The Former Yugoslav Republic of Macedonia, Turkey, United Kingdom.
**Austria, Bulgaria, Czech Republic, Germany, Greece, Hungary, Portugal, Romania, Spain, Switzerland, United Kingdom.
193
General Information
REFERENCE SUBSTANCES, PREPARATIONS AND

SPECTRA OF THE EUROPEAN PHARMACOPOEIA
The reference substances and preparations are selected
and verified batches suitable for use as prescribed in the
European Pharmacopoeia. The European Pharmacopoeia
Commission does not guarantee their use for purposes
other than those prescribed. Each vial supplied contains a
quantity sufficient for the prescribed use.
a DDU (Incoterms 2000) basis, namely, delivered duty

unpaid and insurance included. Where the shipment is
identied below as airport consignment only, the goods
are shipped to the buyer on a CIP (Incoterms 2000) basis,
namely carriage and insurance included.
It is recommended that the products be used as soon as

possible.
The Council of Europe (EDQM) delivers the goods to

the buyer not cleared for import and not unloaded by
any means of transport.
The stability of the contents of opened vials or ampoules

cannot be guaranteed.
It should be noted that no certificates of analysis,
expiry dates, nor any data not relevant to the use of
the products as defined by the Ph. Eur. monograph
are provided with the reference material. The products
comply with the requirements of the monograph and are
monitored regularly.
For complete and daily updated information (e.g.
availability, origin, assigned value, batch validity),
consult the reference substances database on our
website.
CONDITIONS OF SALE
1755 reference substances, reference preparations
and reference spectra are supplied by the Technical
Secretariat of the European Pharmacopoeia Commission.
Prices
PRICE LIST
Prices are identied for each product in the catalogue.

However, please note that prices and package sizes are
subject to change without notice.
The European Directorate for the Quality of Medicines
(EDQM) does not operate a discount policy. The sale
prices are exclusive of duties and taxes and are given
in Euros. It is the responsibility of the buyer (or the
recipient of the delivery if different from the buyer) to
contact the national scal or customs authorities to pay
the duties and taxes. In no event shall the said duties and
taxes be paid by the Council of Europe (EDQM).
In the European Union (EU), there is no VAT identication number for organisations with diplomatic status.
The Council of Europe (EDQM) therefore has no VAT
identication number and is not subject to duties and
taxes.
The goods remain the property of the Council of Europe
(EDQM) until the invoice has been paid in full. Catalogue
items are not returnable for exchange or refund.
DELIVERY AND RELATED COSTS
Unless otherwise stated below or specically agreed with

the customer, the goods are shipped to the buyer on
194
The Council of Europe (EDQM) bears the cost and

risks of packing, transport to the delivery site and
insurance.
In no event shall the Council of Europe (EDQM) be
held responsible for any deterioration of the goods
due to their delayed delivery by the carrier.
The buyer is responsible for the cost of import
customs clearance, for paying the duties and taxes
required in the country of import and for unloading
the goods. The buyer shall be entirely responsible
if the goods are held up at customs at the time of
import into the buyers country. In addition, the
buyer is responsible for any risks associated with use
of their own carrier.
Where the shipping costs are paid by the customer,
the goods are shipped to the buyer on an EX Works
(Incoterms 2000) basis, with neither carriage nor
insurance included. Therefore, the Council of
Europe (EDQM) takes no responsibility in any case of
deterioration or loss of goods.
In no event shall the Council of Europe (EDQM) be
able to provide any assistance.
Delivery charges
The extra charges are applied per shipment. A shipment
comprises only the reference substances that can be
shipped under the same conditions. Consequently, goods
requiring specic packaging (e.g. ice), dangerous goods
or controlled substances will be invoiced separately from
the rest of the order and extra charges will be incurred.
As one order could include several shipments, the
Council of Europe (EDQM) advises its customers to regroup their orders by type of shipment so the customers
can better track the progress of a complete order and save
money in shipping charges.
Where the buyer requests shipping conditions other than
those recommended in our catalogue, or another carrier,
the Council of Europe (EDQM) takes no responsibility in
any case of deterioration of the goods or loss of parcel.
Extra charges (postage and packaging) will be applied in
the following cases (for larger quantities, prices are given
on request). Please note that prices are subject to change
without notice.
General Information
a) Shipment at room temperature

France: no extra charge, price is inclusive of
packaging and postage. At the clients request, Express
Courier delivery is charged at 18 EUR per shipment
20 sales units). For orders of over 100 vials (20 sales

units): prices on request
(Note: for countries outside France, our shipment is
by airport consignment only)
EU: 18 EUR per shipment
Other European countries : 80 EUR per shipment
Other European countries: 250 EUR per shipment
Outside Europe: 120 EUR per shipment
(Note: for India, South America and Africa, our

shipment is by airport consignment only)
Shipping costs paid by the customer: 10 EUR per
shipment
b) Shipment under ice (+ 5 C): sent in cooled freight
containers and by express courier
(Note: for all countries inside the EU (with the
exception of Cyprus, Estonia, Malta, Poland) our
shipment is on a door to door basis. For all other
countries (and the exceptions above), our shipment is
f) Dangerous goods: sent by carrier chosen by EDQM

(Note: for countries outside France, our shipment is
EU: 150 EUR per item
Other European countries: 180 EUR per item
Outside Europe: 250 EUR per item
g) Dangerous goods in excepted quantities: sent by
carrier chosen by EDQM
(Note: for countries inside the EU (with the exception
of Cyprus and Malta), our shipment is on a door to
door basis. For all other countries (and the exceptions
above), our shipment is by airport consignment only)
EU: 100 EUR per item
Other European countries: 125 EUR per item

shipment
Outside Europe: 125 EUR per item
c) Shipment under ice (- 20 C): sent in cooled freight

containers and by express courier
h) Dangerous goods sent by road: carrier chosen by

EDQM

shipment
d) Shipment under dry-ice: sent in cooled freight
containers (dry-ice) and by express courier
shipment
e) Hepatitis C virus BRP, B19 virus DNA for NAT: dry ice
+ dangerous goods from 5 to 100 vials (from 1 to
EU: 150 EUR per item (door to door)

Other European countries: 180 EUR per item (door
to door)
Outside Europe: cannot be sent
i) Precursors (controlled drugs: sent by carrier chosen
by EDQM)
(Note for countries outside the EU, our shipment is by
airport consignment only)
France: no extra charge, price is inclusive of packaging
and postage. At the clients request, Express Courier
delivery is charged at 18 EUR per shipment
NB: these extra charges include packaging, shipping
and management of permits
j) Psychotropic substances (controlled drugs: sent by
carrier chosen by EDQM)
(Note for countries outside France, our shipment is by
France: no extra charge, price is inclusive of packaging
and postage. At the clients request, Express Courier
delivery is charged at 18 EUR per shipment
EU (except France): 110 EUR per shipment
195
General Information
Outside EU: 160 EUR per shipment

k) Narcotics (controlled drugs: sent by carrier chosen by
EDQM)
(Note for countries outside France, our shipment is by
details of the Delivery/Dispatch address (if different)

including name of company, post code, town,
country (please note STREET ADDRESS ONLY, no
P.O. Boxes)
contact name, telephone number, fax number and
e-mail address: an e-mail address is required for order
conrmation and shipping notication purposes
France: 50 EUR per shipment
VAT number (mandatory within the European

Union)
EU (except France): 110 EUR per shipment
your order reference/purchase order reference
Outside EU: 160 EUR per shipment
item order code

l) Reference spectra
ofcial name of the CRS/BRP as set out in the CRS

catalogue
sales/unit quantity
name and account number of the carrier (if you wish

France: France: no extra charge, price is inclusive of
to use your own)
packaging and postage. At the clients request, Express
Courier delivery is charged at 18 EUR per shipment
If orders are received without the ofcial name of the
CRS/BRP and the full item order code (as set out in
the catalogue) the EDQM takes no responsibility for an
incorrect item being dispatched.
shipment.
Unfortunately, we will not be able to process any orders

received without the above information.
How do I order?
SPECIAL DOCUMENTATION is required for controlled drugs and

chemical precursors of narcotics (Vienna Convention)
(see our catalogue for further information).
The reference substances, reference preparations and

reference spectra are supplied by the EDQM.
Payment
ORDER FORM
Please send your order using the CRS order form (see
page xv of the CRS catalogue) or by sending an ofcial
purchase order on company letterhead to the EDQM.
The order form may be downloaded from www.pheur.org
under Reference Substances.
Fax: +33 (0)3 88 41 27 71 for the attention of CRS Sales
E-mail: crs@pheur.org
Payment can be made by cheque made payable to the

Council of Europe/EDQM and sent to the above address
(see 3.1) or by bank transfer to our bank.
Socit Gnrale, 255, route de Mittelhausbergen, 67200
Strasbourg, France
IBAN account number for international transfers:
(FR 76) 30003 02360 00550034256 76
National transfers: 30003 02360 00550034256 76
SWIFT:
SOGEFRPP
You can also pay by credit card (Visa, Eurocard,

Mastercard or American Express) by writing down the
card number, the expiry date and the card holders name,
and including the card holders signature. Please note
Customers are nancially responsible for duplicate orders that we do not accept credit card numbers by telephone.
in the following cases:
In all cases, the payment should be net of charge for
the Council of Europe and invoices should be paid
conrmation orders that are not clearly marked as
within 30 days from the date of invoice. Any other fees,
being a conrmation of an order that has already
been sent to the Council of Europe (EDQM)
such as customs duties, taxes, or tariffs are also the
submission of the same order multiple times (i.e., via responsibility of the customer.
fax, e-mail, mail or any combination thereof)
For certain countries, especially those having strict
monetary regulations, we reserve the right to require
Please note we do not accept orders by telephone.
pre-payment for new clients and large orders. In case of
If you are using any other documentation other than the doubt, please contact us at crs@pheur.org. Payment by
ofcial CRS order form please ensure you have included: letter of credit is not accepted.
Letter:
Council of Europe, European Directorate for

the Quality of Medicines, FAO CRS Sales Team,
BP907, F-67029 Strasbourg Cedex, France
details of the Invoicing/Billing address including

name of company, post code, town, country and
telephone number
196
General Information
Council of Europe
European Directorate for the Quality of Medicines
CRS Order Form

Tel: +33 (0)3 88 41 30 30
BP 907, 67029 Strasbourg Cedex 1 (France)

website: http://www.pheur.org
Helpdesk: http://www.pheur.org/site/page_521.php
SIRET: 778860080010
APE Code APE:990Z
VAT N: Not applicable to Council of Europe - diplomatic privilege.
Fax: + 33 (0)3 88 41 27 71
BILLING ADDRESS
Your Client Code
Company Name*
Invoice Address*
DELIVERY ADDRESS
City*
PostCode*
Country*
City*
PostCode*
Country*
Contact Name*
VAT N(* in Europe)
Tel*
E-mail
Contact Name*
VAT N(* in Europe)
Tel*
Email
(Please complete if different from invoicing address)
Company Name*
Delivery Address*
Fax
Fax
All items marked with an asterisk (*) are mandatory
Your Order Reference* [reference]

Reference*
Date*
Item*
Unit Price
(1) A CRS/BRP may include several individual vials (see sale unit in catalogue), in such instances do not order in terms of total number
of vials.
Quantity(1)*
Total
Total Goods/
DELIVERY CHARGES and PRICES

The price should not be regarded as representing the selling price of a commercial product. The prices quoted in our catalogue are
exclusive of duties and taxes. Extra handling charges may be applied. Please see our catalogue for details.
CONDITIONS OF SALE
We sell on our standard terms of business. For details please see our catalogue.
Payment
I would like to pay now. I will automatically receive an invoice/receipt
I enclose a cheque made payable to Council of Europe/EDQM
I wish to pay by credit card
Visa N
Euro/Mastercard N
Expiry Date
Name
American Express N
Signature
I will pay on receipt on a invoice
197
General Information
ORDER FORM
CATALOGUE OF
- CHEMICAL REFERENCE SUBSTANCES - BIOLOGICAL REFERENCE PREPARATIONS - INFRARED REFERENCE SPECTRA - MISCELLANEOUS REAGENTSThe catalogue of reference substances of the European Directorate for the Quality of Medicines is
a publication of the Council of Europe, issued three times a year to include the latest substances
adopted by the European Pharmacopoeia Commission.
This catalogue is free; if you would like to receive subsequent updated versions, please complete
and return this form.
RECIPIENT:
Please check the appropriate box:
Prof
Dr
Mr
Ms
Surname...................................................................First name...........................................................
Company/Laboratory ..........................................................................................................................
Industrial Laboratory
Private Control Laboratory
Public Control Laboratory
Department:
Invoicing/Purchasing
Analytical Laboratory
Address.................................................................................................................................................
..............................................................................................................................................................
City.............................................................Postal Code .........................Country.................................
Tel.............................................................................Fax......................................................................
ADDITIONAL RECIPIENT IN YOUR COMPANY/LABORATORY:

Please check the appropriate box:
Prof
Dr
Mr
Ms
Surname...................................................................First name...........................................................
Department: Invoicing/Purchasing
Analytical Laboratory
Other service (please describe)...................................................................................
EUROPEAN DIRECTORATE FOR THE QUALITY OF MEDICINES

226, avenue de Colmar - BP 907 - F 67029 Strasbourg Cedex 1, France
Fax +33(0)3 88 41 27 71
198
General Information
LIST OF REFERENCE SUBSTANCES ADOPTED

AT THE NOVEMBER 2005 SESSION OF
THE EUROPEAN PHARMACOPOEIA COMMISSION,
AND NOW AVAILABLE
NEW REFERENCE SUBSTANCES
REPLACEMENT BATCHES
Name
Code
Name
Code
Buspirone hydrochloride CRS

Calcipotriol monohydrate CRS
Cefalotin for impurity B identication CRS
Cefotaxime acid CRS
Cefotaxime sodium
for peak identication CRS
Desogestrel for system suitability CRS
Dihydrotachysterol CRS
Dihydrotachysterol for system suitability CRS
Gembrozil CRS
Glimepiride CRS
Hydromorphone hydrochloride CRS
myo-Inositol CRS
Pamidronate disodium pentahydrate CRS
Rocuronium for peak identication CRS
Spectinomycin hydrochloride CRS
Spectinomycin for system suitability CRS
Sultamicillin CRS
Sultamicillin for peak identication CRS
Trandolapril CRS
Trandolapril impurity C CRS
Trandolapril impurity D CRS
Y0000131
Y0000473
Y0000505
Y0000420
Alfacalcidol CRS 4
Allopurinol impurity C CRS 2
Betadex CRS 4
Carbamazepine impurity A CRS 4
Ceftazidime impurity A CRS 2
4-Epitretracycline hydrochloride CRS 6
Erythromycin C CRS 4
Fenobrate impurity A CRS 3
Flucloxacillin sodium CRS 6
Hydrocortisone CRS 8
Insulin (human) CRS 3
N-Acetyl-cys1-calcitonin CRS 3
Propofol CRS 3
Residual solvents solution class 1 CRS 2
Sodium cromoglicate CRS 2
Terbutaline impurity C CRS 3
A0325450
A0350030
B0950000
Y0000033
C0690510
E0600000
E1320000
F0048005
F0150000
H1300000
I0310000
C0200010
Y0000016
R0250000
S0750000
T0050015
Y0000506
Y0000510
Y0000544
Y0000482
Y0000513
Y0000515
Y0000446
Y0000485
Y0000524
Y0000527
Y0000597
Y0000549
Y0000530
Y0000532
Y0000501
Y0000495
Y0000496
________________________________________________________________________________
COUNTERFEIT MEDICINES - SURVEY REPORT (2006)

Counterfeit medicines pose an ever-greater threat to public health in Europe today. In an effort to adequately
measure the scope of the phenomenon and reduce the inherent risks, the Council of Europe has commissioned a
survey on issues related to this particularly disquieting form of fraud. This report
covers, among others: the current and estimated market and trade matters; the
status of pharmaceutical regulation; national and international co-operation between
authorities, the industry and wholesalers; detection systems and procedures; the
adequacy of legal, judicial and administrative systems; and professional training in the
matter. It also sets out to dene counterfeit medicine and pharmaceutical crime.
Authors: Jonathan Harper, Bertrand Gellie
ISBN: 92-871-5863-0
Price: 25 EUR (Europe) / US$ 38 (outside Europe), + 10 % postage
Available in English only
Council of Europe Publishing - Sales Unit
Ms Sophie Lobey, F-67075 Strasbourg Cedex, France.
Tel: +33 (0)3 88 41 25 81 - Fax: +33 (0)3 88 41 39 10
E-mail: publishing@coe.int - Website: http://book.coe.int
199
General Information
LIST OF TEXTS ADOPTED

AT THE NOVEMBER 2005 SESSION
OF THE EUROPEAN PHARMACOPOEIA COMMISSION
Unless otherwise indicated, these texts will be published in Supplement 5.6 of the European Pharmacopoeia.
Individual copies of texts will not be supplied.
NEW TEXTS
GENERAL CHAPTERS
2.6.27. Microbiological control of cellular products
2.7.23. Numeration of CD34/CD45+ cells in
haematopoietic products
2.7.24. Flow cytometry
2.7.27. Flocculation value (Lf) of diphtheria and tetanus
toxins and toxoids (Ramon assay)
2.9.31. Particle size analysis by laser light diffraction
2.9.33. Characterisation of crystalline and partially
crystalline solids by X-ray powder diffraction
(XRPD)
2.9.43. Apparent dissolution of powders
5.12. Reference standards
5.14. Gene transfer medicinal products for human use
MONOGRAPHS
Vaccines for human use
Anthrax vaccine for human use (adsorbed, prepared from
culture ltrates) (2188)
Diphtheria, tetanus and poliomyelitis (inactivated)
vaccine (adsorbed, reduced antigen(s) content) (2328)
Diphtheria, tetanus, pertussis (acellular, component) and
poliomyelitis (inactivated) vaccine (adsorbed, reduced
antigen(s) content) (2329)
Feline chlamydiosis vaccine (inactivated) (2324)
Mycoplasma gallisepticum vaccine (inactivated) (1942)
Monographs
Alverine citrate (2156)
Benzoin, Sumatra (1814)

Benzoin tincture, Sumatra (1813)
Capsicum oleoresin, rened and quantied (2336)
Capsicum tincture, standardised (2337)
Cascara tincture, standardised (1844)
Cefepime dihydrochloride monohydrate (2126)
Cladribine (2174)
Dembrexine hydrochloride monohydrate for veterinary
use (2169)
Dopexamine dihydrochloride (1748)
Enalaprilat dihydrate (1749)
Gemcitabine hydrochloride (2306)
Guaiacol (1978)
Human haematopoietic stem cells (2323)
Human von Willebrand factor (2298)
Letrozole (2334)
Loratadine (2124)
Mandarin oil (2355)
Manganese glycerophosphate, hydrated (2163)
Milk thistle dry extract, rened and standardised (2071)
Modanil (2307)
Naproxen sodium (1702)
Oxitropium bromide (2170)
Purple coneower herb (1823)
Purple coneower root (1824)
Pyrrolidone (2180)
Terazosin hydrochloride dihydrate (2021)
Tropisetron hydrochloride (2102)
Verbena herb (1854)
REVISED TEXTS
GENERAL CHAPTERS
1.
General notices
2.2.7. Optical rotation
2.2.25. Absorption spectrophotometry, ultraviolet and
visible
2.4.14. Sulphated ash
2.4.22. Composition of fatty acids by gas chromatography
2.6.7. Mycoplasma
2.6.12. Microbiological examination of non-sterile
products (total viable aerobic count)
2.6.13. Microbiological examination of non-sterile
products (test for specied micro-organisms)
2.7.5. Assay of heparin
2.7.20. In vivo assay of poliomyelitis vaccine (inactivated)
5.1.4. Microbiological quality of pharmaceutical
preparations
200
MONOGRAPHS
Dosage forms
Liquid preparations for oral use (0672)
Nasal preparations (0676)
Preparations for irrigation (1116)
Vaccines for human use
Diphtheria and tetanus vaccine (adsorbed, reduced
antigen(s) content) (0647) (previously Diphtheria and
tetanus vaccine (adsorbed) for adults and adolescents)
Diphtheria vaccine (adsorbed, reduced antigen content)
(0646) (previously Diphtheria vaccine (adsorbed) for
adults and adolescents)
Mareks disease vaccine (live) (0589)
Newcastle disease vaccine (inactivated) (0870)
Tetanus vaccine for veterinary use (0697)
General Information
Monographs
Agnus castus fruit (2147)
Ascorbic acid (0253)
Ascorbyl palmitate (0807)
Azithromycin (1649)
Bendroumethiazide (0370)
Betacarotene (1069)
Bisacodyl (0595)
Bromazepam (0879)
Bromocriptine mesilate (0596)
Buserelin (1077)
Calcitonin (salmon) (0471)
Calcium ascorbate (1182)
Cetostearyl alcohol (type A), emulsifying (0801)
Cetostearyl alcohol (type B), emulsifying (0802)
Cholecalciferol concentrate (oily form) (0575)
Cholecalciferol concentrate (powder form) (0574)
Cholecalciferol concentrate (water-dispersible form)
(0598)
Cyanocobalamin (0547)
Diazepam (0022)
Fluconazole (2287)
Glycerol mono-olate (1430)
Human anti-D immunoglobulin (0557)
Human anti-D immunoglobulin for intravenous
administration (1527)
Human coagulation factor VIII (0275)
Human brinogen (0024)
Human normal immunoglobulin (0338)
Human normal immunoglobulin for intravenous
administration (0918)
Human plasma (pooled and treated for virus inactivation)
(1646)
Human plasma for fractionation (0853)

Hydroxocobalamin acetate (0913)
Hydroxocobalamin chloride (0914)
Hydroxocobalamin sulphate (0915)
Levodopa (0038)
Lorazepam (1121)
Magnesium oxide, heavy (0041)
Magnesium oxide, light (0040)
Mannitol (0559)
Methotrexate (0560)
Phytomenadione (1036)
Phytosterol (1911)
Prednisolone acetate (0734)
Riboavin sodium phosphate (0786)
Sodium ascorbate (1791)
Sodium sulphite, anhydrous (0775)
Sodium sulphite heptahydrate (0776)
Stannous chloride dihydrate (1266)
Stearic acid (1474)
all-rac-Tocopherol (0692)
RRR-D-Tocopherol (1256)
all-rac-D-Tocopheryl acetate (0439)
RRR-D-Tocopheryl acetate (1257)
D-Tocopheryl acetate concentrate (powder form) (0691)
DL-D-Tocopheryl hydrogen succinate (1258)
RRR-D-Tocopheryl hydrogen succinate (1259)
Vitamin A (0217)
Vitamin A concentrate (oily form), synthetic (0219)
Vitamin A concentrate (powder form), synthetic (0218)
Vitamin A concentrate (solubilisate/emulsion), synthetic
(0220)
Zuclopenthixol decanoate (1707)R
________________________________________________________________________________
NEW: PHARMEUROPA, PHARMEUROPA BIO AND

PHARMEUROPA SCIENTIFIC NOTES ONLINE
Pharmeuropa, Pharmeuropa Bio and Pharmeuropa Scientic Notes Online are now available as a complementary
service for subscribers to the printed edition of Pharmeuropa, and will be offered for a trial period without an
additional fee for those who ordered the printed version of Pharmeuropa Vol. 18 (2006). All issues stretching back to
volume 10 (1998) are stored as Acrobat PDF les, and can be searched with a search engine identical to the one used
for the online versions of the European Pharmacopoeia and the Standard Terms.
A username and a password are required to access Pharmeuropa Online, and instructions for creating these using the
EDQM Certicate of Authenticity can be found on the inside-front cover of Pharmeuropa 18.1.
201
General Information
UPDATED WORK PROGRAMME OF

THE EUROPEAN PHARMACOPOEIA
(November 2005)
The following new monographs, general chapters and revision items have recently been added to the work
programme of the European Pharmacopoeia.
Interested parties are encouraged to contact the EDQM (lynn.kelso-eleuterio@pheur.org) in order to participate
in the work by provision of data and samples. All information received will be taken into account in monograph
preparation.
Text to be revised
2.2.33. Nuclear magnetic resonance spectrometry: general revision of the chapter to modernise it
2.6.20. Anti-A and Anti-B haemagglutinins: improvement of the method
2.9.9. Measurement of consistency by penetrometry: dimension of apparatus
5.3. Statistical analysis of results of biological assays and tests: Latin square design
Agnus castus fruit / Agnus casti fructus (2147): denition, assay
Cefazolin sodium (0988): related substances
Clove oil (1091): E-caryophyllene content
Cocaine hydrochloride (0073): related substances
Colistin sulphate (0320): loss on drying
Devils claw root / Harpagophyti radix (1095): assay
Ethyl parahydroxybenzoate sodium (2134): heavy metals
Extracts (0765): addition of a section on oleoresins
Folic acid (0067): related substances
Glucagon, human (1635): general revision
Gonadorelin acetate (0827): general revision
Human anti-D immunoglobulin (0557): quality of albumin used as stabiliser
Human anti-D immunoglobulin for intravenous administration (1527): quality of albumin used as stabiliser
Levothyroxine sodium (0401): chiral purity
Liquid preparations for cutaneous application (0927): uniformity of dosage units
Melissa leaf / Melissae folium (1447): assay
Minocycline hydrochloride (1030): denition, degree of hydration
Nicotinamide (0047): related substances
Phytomenadione (1036): related substances
Protamine hydrochloride (0686): test for absorbance
Rectal preparations (1145): uniformity of dosage units
Roselle / Hibisci sabdariffae os (1623): identication
Semi-solid preparations for cutaneous application (0132): uniformity of dosage units
Thiamine hydrochloride (0303): related substances
Thiamine nitrate (0531): related substances
Vaginal preparations (1164): uniformity of dosage units
Willow bark / Salicis cortex (1583): assay
New texts to be elaborated

Cholesterol for parenteral use (2397)
Cod-liver oil, farmed (2398)
Microcrystalline wax (2399)
Parafn, synthetic (2263)
Trimagnesium citrate 9-hydrate (2402)
Trimagnesium citrate 14-hydrate (2401)
202
General Information
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i]Za^hid[gZ[ZgZcXZhjWhiVcXZhjhZY^ci]Zbdcd\gVe]
i]Za^hid[XZgi^[^XViZh\gVciZY#
________________________________________________________________________________
LIST OF CODES OF GROUPS OF EXPERTS

(November 2004)
Groups of experts
1
6
6B
7
10A
10B
10C
10D
Microbiology
Biological substances
Human blood and blood products
Antibiotics
Organic chemistry - synthetic products
Organic chemistry - synthetic products
Organic chemistry - synthetic Products
Organic chemistry - synthetic Products
11
12
13A
13B
13H
14
15
15V
Organic chemistry - natural products

Galenical products
Phytochemistry
Phytochemistry
Fatty oils and derivatives
Radioactive compounds
Sera and vaccines
Veterinary sera and vaccines
Working parties
BOT
BSR
CEL
CRB
CTP
FRC
GEL
GTP
HFA
HOM
ICP
INC
Botulinum toxin
Bovine serum
Cellulose derivatives
Carbohydrates
Cell therapy products
Functionality-related characteristics
Gelatin
Gene therapy products
Propellants
Homeopathy
Inductively coupled plasma spectrometry
Inorganic chemistry
INH
LEC
MAB
MMM
MYC
P4
POW
RGN
ST
STA
VIT
WAT
Inhalations
Lecithins for pharmaceutical purposes
Monoclonal antibodies
Alternative microbiological methods
Mycoplasmas
Procedure 4
Powder characterisation techniques
Reagents
Standard terms
Statistics
Vitamins
Water
203
General Information
LIST OF STANDARD TERMS

5th EDITION (printed version available)
(27 European languages)
The present list of Standard Terms is a revised list that was drawn up in response to a request from the European
Commission. It covers medicines for both human and veterinary use. These Standard Terms are to be used in
answering the questions in Module 1 (items 1.2 and 1.3) of the EU application form.
The list of Standard Terms is composed of:
an introduction:
a section of general principles and instructions for the use of Standard Terms,
the summary of the changes (amendments, additions, deletions) performed since the last publication
(December 2002),
the procedure for the addition, deletion or modification of terms in the list of Standard Terms (requests
restricted to licensing authorities);
3 lists of standard terms:
list of pharmaceutical forms,
list of routes and/or methods of administration,
list of containers, closures and administration devices.
The 5th Edition contains translations in 27 European languages: Bulgarian, Croatian, Czech, Danish, Dutch, English,
Finnish, French, German, Greek, Hungarian, Icelandic, Italian, Macedonian, Norwegian, Polish, Portuguese, Slovak,
Slovenian, Spanish, Swedish and Turkish. 5 new languages have been added compared to the printed version
published in December 2002: Estonian, Latvian, Lithuanian, Maltese and Romanian.
The corresponding online version is available only to people who ordered the printed version of the 5th Edition
(December 2004).
Price: see the catalogue on our website http://book.pheur.org
________________________________________________________________________________
NEW AND REVISED STANDARD TERMS

These new or revised standard terms have been recently adopted; once their translation into the 26 other languages is
available, they will be included in the online version of the standard terms.
NEW STANDARD TERMS
DEFINITION
Intraosseous use
Administration of a medicinal product into the bone marrow.

Intrasternal use is excluded.
Medicated thread
Gastric use
Administration of a medicinal product to the stomach by means of an

appropriate device
REVISION OF EXISTING STANDARD

TERM
DEFINITION
Gastroenteral use
Administration of a medicinal product to the stomach or duodenum

gastroenteral tract by means of an appropriate device. For use only
when gastric use and intestinal use do not apply
204
General Information
ELABORATION / REVISION OF A MONOGRAPH

(Procedure 1)
The European Pharmacopoeia Commission decides to
elaborate/revise a monograph
j
Group of experts:
a rapporteur prepares a draft monograph, which is
evaluated by the experts
j
j
Pharmeuropa (4 issues per year):

the draft monograph is published for public enquiry,
which lasts 3 months
j
The national pharmacopoeia authorities process the
comments received on the draft
The EDQM-Division 1 compiles the comments sent by

the national authorities
The revised draft is

published for further
enquiry, if necessary
j
j
The group of experts examines the comments and

revises the draft monograph accordingly
j
The draft is proposed to the
Commission
j
European Pharmacopoeia Commission
j
- adopts the monograph, if necessary with slight
modications
- adopts the implementation date (about 1 year
after the adoption of the monograph)
j
does not adopt the monograph
j
EUROPEAN PHARMACOPOEIA (3 supplements per year):
the monograph is published about 6 months after adoption
205
General Information
ETHICAL EYE - BIOMEDICAL RESEARCH (2004)

What are the rules and underlying values governing biomedical research in Europe? What form do these values take
and where do they originate? Does biomedical research pose a threat to individuals and their rights? What balance
should be struck between freedom of research and protection of the individual? All these questions are examined in
this book from a pan-European perspective.
The authors look at various international and European standards, including the Helsinki Declaration of the World
Medical Association, EU Directive 2001/20 on pharmaceutical research and the Council of Europes Convention on
Human Rights and Biomedicine. The last named was signed in Oviedo in 1997 and is the rst binding international
treaty on the subject, with a special chapter on scientic research on human beings. The Convention establishes a
common minimum level of protection of fundamental rights throughout Europe. It will soon be supplemented by an
additional protocol specically concerned with biomedical research.
The book contains a glossary and a list of relevant international conventions and treaties, websites and publications.
It is aimed at both specialists and a wider public interested in this subject.
Contents
BIOMEDICAL RESEARCH IN EUROPE
Preface
Introduction by Claude Huriet (France)
History and denitions by Povl Riis (Denmark)
Germany: current legislation by Jochen Taupitz (Germany)

Central and eastern Europe: research-related problems for
transition countries by Eugenijus Gefenas (Lithuania)
Italy: some shortcomings of biomedical research by Stphane
Bauzon (Italy)
United Kingdom: data protection and condentiality by
Michel Coleman and Vivienne Harpwood (United Kingdom)
ETHICAL DILEMMAS IN RESEARCH

Uses and abuses of biomedical research
by Jan Helge Solbakk (Norway)
Selection and recruitment of participants: European standards
by Herman Nys (Belgium)
Placebo: its action and place in health research
by Andrzej Grski (Poland)
Cancer clinical trials by Maxime Seligmann (France)
Some ethical considerations in industry-sponsored clinical
trials by Tom Gallacher and N. Sreeharan (United Kingdom)
Women in biomedical research by Outi Leena L. Hovatta
(Sweden)
EUROPE AND BIOMEDICAL RESEARCH

European law and biomedical research by Peteris Zilgalvis
(Council of Europe)
APPENDICES
Selected websites
Draft protocol on biomedical research, Council of Europe
The Helsinki Declaration, World Medical Association
ISBN 92-871-5462-7. Price: 15 EUR (Europe) / US$ 23 (outside Europe), + 10 % postage.

This book is available in French and English from:
Ms Sophie Lobey, F- 67075 Strasbourg Cedex, France.
Tel: +33 (0)3 88 41 25 81 - Fax: +33 (0)3 88 41 39 10
E-mail: publishing@coe.int - Website http://book.coe.int
________________________________________________________________________________
NEW: TECHNICAL GUIDE FOR THE

ELABORATION OF MONOGRAPHS - 4th Edition (2005)
now available as a free download on the EDQM website
The Technical Guide for the Elaboration of Monographs describes the scientific approach used for the elaboration of
monographs and the establishment of specifications of the European Pharmacopoeia. The guide also describes how
to scientifically elaborate the various sections that must be included in each monograph, for example, definition,
characters, the physical and chemical reactions constituting the identification section, purity tests, assay methods
and storage conditions. It is continually being updated.
Available in English and French as a free download from the EDQM website http://www.pheur.org
206
General Information
GUIDE TO SAFETY AND QUALITY ASSURANCE

FOR ORGANS, TISSUES AND CELLS
2nd Edition (2004)
The purpose of this guide is to provide guidance for all those involved in transplantation to maximise the quality,
and thereby the success rate, of transplants, and to minimise the risks to all involved in this complex procedure. It
includes safety and quality standards for the procurement, preservation, processing and distribution of organs, tissues
and cells of human origin (allogeneic and autologous) used for transplantation purposes. This guide will be regularly
updated, in line with the latest technical advances.
As the European Union Directive on Tissues and Cells (2004/23/EC) was recently adopted, the European Commission
will build on the Council of Europes guide when establishing technical standards under the directive. This
co-operation will ensure that the same standards are applied throughout Europe.
The 2nd Edition of the Guide can be obtained in English and French from:
Tel: +33 (0)3 88 41 25 81 - Fax : +33 (0)3 88 41 39 10
Questions and comments on the content should be sent directly to the division in charge:
Council of Europe, Health Division
Mr Karl-Friedrich Bopp, F-67075 Strasbourg Cedex, France.
Tel: +33 (0)3 88 41 22 14 - Fax: +33 (0)3 88 41 27 26
E-mail: karl-friedrich.bopp@coe.int
Council of Europe website: www.coe.int
Health and Ethics: www.coe.int/T/E/Social_Cohesion/Health
________________________________________________________________________________
VISIT OUR WEBSITE http://www.pheur.org

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download about 1600 safety datasheets and about 190 leaflets
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NEW: the reference substances database is now updated daily with information on availability; other accessible
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find official surveys in progress, announcements of conferences and international seminars, etc.
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207
General Information
EDQM CONFERENCE PROCEEDINGS

The following free Conference Proceedings are available to download from the EDQM website
Certicates of Suitability of Monographs of the
European Pharmacopoeia: Implementation of the
5th Edition New Procedures for Revision and
Renewal of Certicates
Quality on the Move: Dynamics of the European

Pharmacopoeia
4-6 October 2004, Budapest, Hungary
Process Analytical Technologies International Symposium
3-4 May 2004, Cannes, France
27-28 October 2005, Istanbul, Turkey
OMCL Information Day: Place and Role of the European Microbiological Control Methods in the European
Pharmacopoeia: Present and Future
OMCL Network within the Regulatory Framework in
Europe
5-6 May 2003, Copenhagen, Denmark
27 May 2005, Rome, Italy
Foot and Mouth Disease Vaccines: Current Situation

17-18 March 2003, Strasbourg, France
Alternatives to Whole Cell Pertussis Vaccine Potency

Assay
Standardisation and Quality Control Cell and Gene

Therapy Products
24-25 February 2003, Strasbourg, France
16 March 2005, Geneva, Switzerland
Quality of Homoeopathic Products in the New European

Replacement, Reduction and Renement of the Use of
Legislative Framework
Animals in the Quality Control of Vaccines
15 February 2005, Strasbourg, France
7-8 November 2002, Strasbourg, France
Serological Potency Tests for Diphtheria and Other
Vaccines
Excipients: Classical Requirements and Functionality

Related Testing
4-5 April 2002, Brussels, Belgium
6-7 October 2004, Budapest, Hungary
******
The following Conference Proceedings can be ordered from the EDQM. For prices and ordering please consult the
catalogue on our website http://book.pheur.org
Certication of Suitability of Monographs of the
European Pharmacopoeia (CEP) - New Developments
of the Procedure, How to Apply for a CEP
8-9 November 2001, Athens, Greece
The Future Face of the European Pharmacopoeia Current Concerns in Pharmaceutical Analysis
8-9 February 2001, Cannes, France
Herbal Medicinal Products: Quality Evaluation Contribution of the European Pharmacopoeia
16-17 November 2000, Nice, France
Tetanus Vaccine for Human Use
22-23 June 2000, Strasbourg, France
Mycoplasma Testing: The Potentialities and Role
of PCR Tests
13-14 March 2000, Paris, France
Biologicals beyond 2000: Challenge for Quality

Standards in an Evolving Field
27-29 September 1999, Strasbourg, France
General Monographs on Dosage Forms and PharmacoTechnological Test Methods
26-27 October 1998, Seville, Spain
The Vision of the European Pharmacopoeia in the
21st Century - The Dynamics of Quality of Medicines in
Europe
4-7 December 1996, Prague, Czech Republic
Sterility Tests and Efcacy of Antimicrobial
Preservation
5-6 February 1996, Barcelona, Spain
All above proceedings are available in English only and are not included in the subscription to Pharmeuropa.
208
General Information
INSPECTIONS AND AUDITS

Success factors and improvement prospects
38th International Seminar
7 & 8 June 2006 - Montpellier, France
THE HEALTH INDUSTRY PROFESSIONALS ANNUAL EVENT
Quality assurance & control / regulatory affairs / production / supply chain / R & D etc.
IDENTIFY AND BUILD UP YOUR STRENGTHS
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News and prospects
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Web: www.sfstp.org
________________________________________________________________________________
PHARMEUROPA SCIENTIFIC NOTES

As from Pharmeuropa 17.4, Scientic Notes are principally published in a new publication called Pharmeuropa
Scientic Notes.
Articles published in Pharmeuropa Scientic Notes are indexed in the PubMed database of the National Library of
Medicine, available online (www.ncbi.nlm.nih.gov).
The rst edition of Pharmeuropa Scientic Notes (Pharmeuropa SN 2005-1) became available in August 2005.
This issue is included in the subscription to Pharmeuropa, and is not available separately.
SCIENTIFIC NOTES 2005-1

Few Bicyclic Acetals at Reducing End of LowMolecular-Weight Heparins: Might they Restrict
Specication of Pharmacopoeia?
The Control of Impurities in Chlortalidone
Using a Reversed-Phase Stationary Phase
Development of an in vivo Test Procedure for the Ease

of Breaking of Scored Tablets
Chromogenic Assay of Human Coagulation Factor VIII:
Statistical Comparison of 2 Working Dilution
Procedures
Factor VIII Test in Reference Preparations:

Compensation for Different Dilutions
Impurity Profile of Amino Acids?
The Control of Impurities in Amitriptyline

Hydrochloride Using a Reversed-Phase
Hybrid Stationary Phase
Quality Criteria of Homoeopathic Mother Tinctures:

Considerations Regarding Suitable Tests for
Homoeopathic Monographs
A Precise Colour Determination Method for Tablets an Application of Instrumental Colour Measurement
in the Pharmaceutical Development
Instructions for Authors
Batch Variability of Bacitracin: HPLC versus MEKC
Available now (English only)
For prices and ordering information please consult the catalogue on our website http://book.pheur.org
209
General Information
PHARMEUROPA BIO
These issues are included in the subscription to Pharmeuropa.
BIOLOGICALS 2005-1
Collaborative Study to Establish a New Biological
Reference Preparation for Prekallikrein Activator
Collaborative Study for the Establishment of the Ph.
Eur. BRP Batch 1 for Anti-Vaccinia Immunoglobulin
Feasibility Study to Develop a Common in vitro
D-Antigen Assay for Inactivated Poliomyelitis Vaccines
Allergy Vaccines: a Need for Standardisation in Mass
Units of Major Allergen
Efficacy Demonstration of Tetanus Vaccines by double
antigen ELISA
International Symposium on Alternatives to Whole Cell
Pertussis Vaccine Potency Assay
Collaborative Study for the Validation of Serological

Methods for Potency Testing of Diphtheria Toxoid
Vaccines: Part 1
Collaborative Study for the Validation of Serological
Methods for Potency Testing of Diphtheria Toxoid
Vaccines: Extended study: Correlation of Serology with
In Vivo Toxin Neutralisation
Establishment of European Pharmacopoeia Biological
Reference Preparations Batch 2 for rDNA Hepatitis B
vaccine (Method A and B)
Control of Clostridium Perfringens Vaccines by Means
of an Indirect Competitive ELISA for the Epsilon
Toxin Component Examination of the Assay by a
Collaborative Study
BIOLOGICALS 2004-1
BIOLOGICALS 2003-1
Validation Study to Evaluate the Reproducibility of a

Candidate In Vitro Potency Assay of Newcastle Disease
Vaccines and to Establish the Suitability of a Candidate
Biological Reference Preparation
Establishment of Batch 4 of the Biological Reference
Preparation (BRP) for Rabies Vaccine (Inactivated) for
Veterinary Use
Collaborative Study for the Establishment of
Erythropoietin BRP Batch 2
Somatropin and its Variants: Structural
Characterization and Methods of Analysis
Capillary Electrophoresis for the Control of Impurities
of rDNA Somatropin
Collaborative Study to Establish the Low-MolecularMass Heparin for Assay European Pharmacopoeia
Biological Reference Preparation (BSP060)
Collaborative Study for the Establishment of The

European Pharmacopoeia BRP Batch 1 for
Diphtheria Toxin
Collaborative Study for the Establishment of
European Pharmacopoeia BRP Batch 2 for Inactivated
Poliomyelitis Vaccine for In Vitro D Antigen Assay
Collaborative Study for the Establishment of A Global
(WHO International / US/Ph. Eur.) Standard for the
Potency Assay of Human Anti-D Immunoglobulin
Feasibility Study to Evaluate the Correlation Between
Results of a Candidate In Vitro Assay and Established
In Vivo Assays for Potency Determination of Newcastle
Disease Vaccines
BIOLOGICALS 2003-2
Collaborative Studies for the Establishment of
Reference Substances for the Microbiological Assay of
Antibiotics
Collaborative Study for Establishment of a Global
Standard for the Potency Assay of Human Anti-D
Immunoglobulin
Collaborative Study for Establishment of a European
Pharmacopoeia Biological Reference Preparation (BRP)
For B19 Virus DNA Testing of Plasma Pools by Nucleic
Acid Amplication Technique
BIOLOGICALS 2002-1
Collaborative Study for Establishment of an HPLCMethod for Batch Consistency Control of Recombinant
Interferon-Alfa-2
Calibration of European Pharmacopoeia BRP Batch 3/
Mega 2 (US/FDA) Standard for Human Coagulation
Factor VIII Concentrate for Use in the Potency Assay
Collaborative Study for the Establishment of the
European Pharmacopoeia BRP for Oral Poliomyelitis
Vaccine (OPV) Batch 3 for Use in the Potency Assay
Establishment of the European Pharmacopoeia BRP for
Hepatitis A Vaccine Type B (Aventis Pasteur) Batch 2
For prices and ordering information please consult the catalogue on our website http://book.pheur.org
210
General Information
GUIDE TO THE PREPARATION, USE AND QUALITY

ASSURANCE OF BLOOD COMPONENTS - 12th Edition (2006)
In the absence of substitutes, the use of blood components remains essential in therapy. This guide contains a
compendium of measures designed to ensure the safety, efcacy and quality of blood components and is particularly
intended for all those working in blood transfusion services. In accordance with the approach recommended by the
Council of Europe in this eld, it is based on the premise of voluntary, non-remunerated blood donation. It describes
the different blood components and gives information on their clinical indications and possible side effects. This
guide continues to be the golden standard for blood transfusion services and forms the basis for many national
guidelines in Europe and around the world. For example, in 2000 Australia mandated the guide in its standard for
blood components. During the elaboration of this 12th Edition, the Council of Europe and the European Commission
have worked closely together to ensure that the requirements set under Article 29 of the European Union Directive
2002/98/EC are compatible with those of this guide. Where necessary, chapters have been revised to take into account
the new technological advances. The Guide to the preparation, use and quality assurance of blood components will
be of interest to blood transfusion centres, legislators, health personnel and to all those working in the eld of blood
transfusion.
The European Pharmacopoeia monograph on human plasma for fractionation refers inter alia to the
recommendations made in this Guide.
The 12th Edition of the Guide can be obtained in English and French from:
Tel: +33 (0)3 88 41 25 81 - Fax: +33 (0)3 88 41 39 10
Questions and comments on the content should be sent directly to the division in charge:
Council of Europe, Health Division
Mr Karl-Friedrich Bopp, F-67075 Strasbourg Cedex, France.
Tel: +33 (0)3 88 41 22 14 - Fax: +33 (0)3 88 41 27 26
E-mail: karl-friedrich.bopp@coe.int
Council of Europe website: www.coe.int
Health and Ethics: www.coe.int/T/E/Social_Cohesion/Health
NEWS
EUROPEAN PHARMACOPOEIA
5th Edition of the list of Standard Terms (Dec. 2004)

Printed and online versions available
The present list of Standard Terms is a revised list that was drawn up
in response to a request from the European Commission. It covers
medicines for both human and veterinary use. These Standard Terms
are to be used in answering the questions in Module 1 (item 1.2 and
1.3) of the EU application form.
The publication contains 3 lists of Standard Terms:
the list of pharmaceutical forms,
the list of routes and/or methods of administration,
the list of containers, closures and administration devices.
The 5th Edition contains translations in 27 European languages. New
languages that have been added since 2002 are: Estonian, Latvian,
Lithuanian, Maltese and Romanian.
With every printed publication of the 5th Edition ordered, access is
granted FREE to the online version.
How to purchase
this publication
All EDQM publications may be
ordered quickly and easily using
our online bookshop:
http://book.pheur.org
Ordering online is entirely secure
and helps us to process your order
more efciently.
Other ways to order include:
by fax: 00 33 (0) 3 88 41 27 71
by telephone: 00 33 (0) 3 88 41 30 30
by e-mail: orders@pheur.org
by surface mail:
European Directorate for the Quality of Medicines (EDQM)
Council of Europe
226 avenue de Colmar BP 907
F- 67029 STRASBOURG Cedex 1, FRANCE
For more information, visit our internet site:

http://www.pheur.org
211
General Information
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212
AGENDA 2006
TRAINING SESSIONS ON THE 5th EDITION
OF THE EUROPEAN PHARMACOPOEIA
CHEMICALS NEW PROGRAMME
Chicago, USA, 27-28 April 2006
NEW SESSION ANNOUNCED: Dublin, Ireland, 13-14 November 2006
***
The EDQM is pleased to announce a new training programme on the European Pharmacopoeia 5th Edition. The
new programme aims to provide professionals with an in-depth and up-to-date knowledge of the most important
and practical aspects of the European Pharmacopoeia. New additions for 2006 include practical examples and case
studies, question and answer sessions giving you the opportunity to clarify any issues that may arise and the chance
to meet one-to-one with the speakers who work in a certain area, thus providing you with more meaningful and
worthwhile interactions.
Do not miss these opportunities to meet the EDQM/European Pharmacopoeia.
More information will be available on the EDQM website www.pheur.org and in the next issues of Pharmeuropa.
***
INTERNATIONAL CONFERENCES & SYMPOSIA
NEW! TWO SYMPOSIA ANNOUNCED FOR 2006
Impurities Control: Setting Specications for Antibiotics and Peptides
Strasbourg, France, 21-22 September 2006
-----------------------------------------------------------New Microbiology Chapters of the European Pharmacopoeia
Strasbourg, France, 2-3 October 2006
GENERAL CONDITIONS FOR REGISTRATION AT THE EDQM CONFERENCES

AND TRAINING SESSIONS
HOW TO REGISTER
Register as soon as possible: places are limited.
We recommend using a separate form for each participant. Please ll in the registration form* and return it to
the EDQM Public Relations Unit, by post, by fax or via the HelpDesk duly completed with the selected method of
payment.
NEW Online Registration: Online conference registration is now possible. To use the online registration form, just
click on the icon EDQM Events - Register online and follow the steps described.
REGISTRATION FEE
A special rate is given to permanent staff of national authorities, university, R&D public centres.
The registration fee is not subject to VAT and covers attendance at the lectures, working documents, lunches,
coffee breaks and the ofcial dinner.
The registration fee does not include your hotel accommodation costs (see hotel reservation form) and travel
expenses.
METHOD OF PAYMENT
You can make your payment: by bank transfer, by enclosed cheque or by credit card.
The relevant bank transfer information is given on the invoice.
CANCELLATION POLICY
One month before the event 80% of the registration fee will be refunded; no refunds will be given after this date.
However, registration may be transferred to another person at any time. In case of no show, the registration fee is
due.
* All information requested on the registration form is necessary and used only for the organisation of the seminar.
213
27-28 April 2006 Location: Radisson Hotel, 160E Huron St., Chicago, USA
NEW PROGRAMME: TRAINING SESSION 5TH EDITION
2006 TRAINING SESSION: UNITED STATES

THE EUROPEAN PHARMACOPOEIA 5TH EDITION
CHEMICAL DRUG PRODUCTS AND SUBSTANCES
Duration: 2 days, location: Radisson SAS, 160E Huron Street, Chicago, USA
Working language: English
***
NEW PROGRAMME
THURSDAY 27 APRIL 2006
8:45-9:30 Registration and Welcome Coffee
9:30-9:45 Opening remarks and general introduction
9:45-10:10 Dr Agns Artiges, Director of the European Directorate for the Quality of Medicines,
EDQM, Council of Europe
European regulations for medicines: How does the system work? Relationship between EU/EMEA
and the EDQM of the Council of Europe. Place and roles of the EDQM and the European
Pharmacopoeia. General organisation of the EDQM.
10:10-10:30 Dr Claude Coune, Head of Publications Division, EDQM, Council of Europe
Elaboration and revision of the European Pharmacopoeia.
10:30-11:00 Coffee Break
11:00-11:45 Dr Emmanuelle Charton, Scientific Officer, EDQM, Council of Europe
How to use the European Pharmacopoeia: Understanding the general notices, general chapters,
general monographs and monographs on dosage forms. How to use them in practice.
11:45-12:15 Open discussion with the speakers
12:15-13:30 Lunch Break
13:30-14:15 Dr Emmanuelle Charton
Cases studies of specific monographs (active substances and excipients), use of reference standards
How to use the general monograph Substances for pharmaceutical use to control impurities and
decision trees for impurities. How to interpret chromatograms and list of impurities.
15:00-15:20 Open discussion with the panel of speakers
15:20-15:40 Coffee Break
15:40-16:25 Dr Andrea Lodi, Deputy Head of the Laboratory Division, EDQM, Council of Europe
Identification of the need and uses of a reference standard. Overview of the policy and process used
to establish of a reference standard
17:00-18:00 One to One Consultations
Subject to a prior appointment, questions has to be sent prior the session to guarantee an efficient
answer/advice. Topics under which a consultation can be arranged: General questions on the EDQM,
the European regulatory framework and harmonisation; Technical questions on PhEur monographs
and texts; Reference standards; Electronic version of the European Pharmacopoeia
214
FRIDAY 28 APRIL 2006
27-28 April 2006 Location: Radisson Hotel, 160E Huron St., Chicago, USA
NEW PROGRAMME: TRAINING SESSION 5TH EDITION
8:00-8:45 Welcome Coffee

8:45-9:30 Dr Claude Coune
How we build a monograph
How to successfully submit a monograph?
10:15-10:30 Open discussion with the speakers
10:30-11:00 Coffee break
11:00-11:45 Dr Andrea Lodi
How to use the European Pharmacopoeia in your daily laboratory activity: Case study. Verification
and/or validation of test procedures. System suitability requirements. Reagent and reference
standards. Alternative methods.
11:45-12:00 Open discussion with the speaker
12:00-12:45 Ms Fiona Gilchrist, Public Relations Unit, EDQM, Council of Europe
EDQM Internet sites: Features, which will help you, conduct your regulatory surveillance. How to
make the best use of the online services, specialised databases and the new users support: the
HELPDESK.
12:45-13:00 Open discussion with the speaker
13:00-14:00 Lunch break
14:00-14:20 Dr Agns Artiges
Pharmacopoeias and international harmonisation process
14:20-15:00 Dr Claude Coune
The European Pharmacopoeia Publications (printed and electronic publications).
15:00-15:45 Dr Agns Artiges
The certification procedure and inspections: General considerations, How to apply for a Certificate.
Revisions and renewals in the procedure of certification.
16:00 Final addresses and Closure of the meeting
16:15-17:15 One to One Consultations
Topics covered: General questions on the EDQM, the European regulatory framework and
harmonisation; Technical questions on PhEur monographs and texts; Certification procedure;
Publications and services; Electronic version of the European Pharmacopoeia
During coffee breaks, the electronic/online version of the European Pharmacopoeia will be
set up and demonstrated to participants interested in learning more about this version.
Who should attend?
This conference should be attended by professionals from industry, in particular persons involved in
the manufacture and control of drug substances/products or the preparation of registration dossiers;
from inspectorates, regulatory agencies and academic institutions.
Contact details:
Further information on the training course, accommodation and registration forms will be made
available on the EDQM internet site: http://www.pheur.org. Register at:
http://www.pheur.org/site/page_644.php or contact the Public Relations Unit, EDQM; 226 avenue
de Colmar, BP907, F-67029, Strasbourg, Cedex 1, FRANCE; Tel: 00 33 3 88 41 30 30 (Dial 4);
Fax: 00 33 3 88 41 27 71; Email: Via the EDQM Helpdesk - go to:
215
Please complete and send this form to the Public Relations Unit: By fax: +33 3 88 41 27 71, or via the EDQM
HELPDESK http://www.pheur.org/site/page_521.php: Go to the topic Conferences, Events, Public Relations
and read the question May I send my registration form via the EDQMs internet site for further instructions, or
alternatively register ONLINE, click on Events Register Online and select the name of the event.
27-28 April 2006 Radisson Hotel, 160E Huron St., Chicago, USA
REGISTRATION FORM:
TRAINING SESSION 5th EDITION
REGISTRATION DETAILS
DATE OF REGISTRATION (DD/MM/YY) :
_ __ __ _
REGISTRATION FEE (*See general conditions)

950 * or 400 *
Yes, I am interested in reserving a meeting
ONE TO ONE MEETING
Which topic(s)?
EU Regulations Monographs, revisions
No, I am not interested
Publications Certification
Internet
PARTICIPANT DETAILS Please complete one form per participant

Title (Dr., Mr, Mrs, Ms, )
First Name
Family Name
Company/Institution
Address for
Correspondence
Postcode
Town
Country
Telephone
Fax
E-mail
AREA OF ACTIVITY/ Manufacturer of raw material
Manufacturer of medicines
OCCUPATION:
QA
QC
Regulatory authority:
Licensing
Inspection
OMCL
Other (please specify)
for human use for veterinary use

R&D
Regulatory affairs
Pharmacopoeias
University
PAYMENT (NEW)
Following receipt of your registration form, we will send you an invoice. Please note that we must receive payment
before the conference takes place. Details of payment methods will be outlined on the invoice. However, you will
be able to settle your invoice by:
1. PERSONAL OR COMPANY CHEQUE made payable to Council of Europe/EDQM
2. BANK TRANSFER
3. CREDIT CARD
DETAILS FOR INVOICING PURPOSES (if different from participant details)
Company/Institution
Address
Postcode
Town
Country
VAT Number (EU only)
Contact Name
Telephone
Fax
E-mail
PO Number/ Reference
CANCELLATION CHARGES: I have read and accept the cancellation terms as stated on our website.
Date
Signature
FOR MORE INFORMATION PLEASE VISIT REGULARLY THE WEBSITE : http://www.pheur.org
216
27-28 April 2006 Location: Radisson Hotel, 160E Huron Street, Chicago, USA
ONE-TO-ONE RESERVATION FORM:
2006 TRAINING SESSION: UNITED STATES

EUROPEAN PHARMACOPOEIA 5TH EDITION CHEMICALS
27-28 APRIL 2006
ONE TO ONE CONSULTATION QUESTION FORM

If you would like to register for a one-to-one consultation (15 minutes) with a member of the EDQM
scientific team during the training course, please complete this consultation request form and send it
to the EDQM by fax to +33 3 88 41 27 71 or e-mail via the Helpdesk on the website:
Priority will be given to those who book in advance.
PARTICIPANT DETAILS
Title (Dr., Mr, Mrs, Ms)
First Name
Family Name
Company/Institution
E-mail
The time slots available for one-to-one consultations will be limited. The EDQM shall do all it can to
accommodate all interested participants.
If it is on a specific application, please mention the reference number
Your question:
Date
Signature
FOR MORE INFORMATION ABOUT TRAINING SESSIONS AND CONFERENCES ORGANISED BY THE EDQM
PLEASE VISIT REGULARLY THE WEBSITE : http://www.pheur.org
217
HOTEL RESERVATION FORM

2006 TRAINING COURSE CHICAGO: 27-28 APRIL 2006
27-28 APRIL 2006 Location: Radisson Hotel, 160E Huron St. Chicago, USA
HOTEL REGISTRATION FORM:
Location: Radisson Hotel & Suites Hotel, Chicago, USA

To be filled in and sent by fax before 27 MARCH 2006
Fax: +1-312-757-5158
Global reservation:
40 rooms reserved from Wednesday 26th April to Friday 28th April included.
Contact reservations: RADISSON HOTEL & SUITES: 160E Huron St., Chicago, IL 60611
Contact: Reservations and indicate you are part of the EDQM/European Pharmacopoeia training course;
Tel: +1-312-787-2900; Fax: +1-312-757-5158; E-mail: radchicago@ihrco.com
General information: The quotas of rooms reserved will be available until 27 March 2006.
After this date the availability and the negotiated prices will not be guaranteed. The rooms should be reserved
individually by each participant and are available on a first come, first served basis. Cancellation policy: Before
26 April 2006 at no further cost, after this date and in case of no show, one night will be charged to your credit
card.
Participant:
Family name:
First name:
Address:
City:
Postcode:
State/Country:
Tel:
Fax:
E-mail:
Hotel reservation:
Arrival day/hour:
Departure day/hour:
Please tick the box indicating the type of room you wish to reserve:
Standard Single Room at 149USD per night*
Standard Double Room at 149USD per night*
Single Corner Suite at 189USD per night*
Double Corner Suite at 189USD per night
*Note: Room rates are subject to state and local taxes and excludes breakfast
Please tick the box(es) indicating number of nights accommodation you require:
Yes Night 26 April
Yes Night 27 April
Yes, Night 28 April
Method of payment:
Credit card: Visa
EuroCard/ MasterCard
Amex
Credit card number:
Diners Card
Expiry date:
Cardholder: ______________________________________________________
I authorise the RADISSON HOTEL & SUITES to charge against my credit card the amount equivalent to one
night in order to block my reservation for the duration of the training session.
HOTEL CONFIRMATION RESERVATION N: ____________ Signature: ______________________
218
NEW! 2006 SYMPOSIUM: STRASBOURG

IMPURITIES CONTROL: SETTING SPECIFICATIONS FOR
ANTIBIOTICS AND PEPTIDES
21-22 September 2006
Duration: 2 days, Location: Strasbourg, France
***
FIRST ANNOUNCEMENT
The European Pharmacopoeia has developed a general policy on impurities control set out in the general
monograph Substances for Pharmaceutical Use and the general chapter 5.10 Control of impurities in
substances for pharmaceutical use. Antibiotics and peptides are not covered by this general policy and the
main aim of this symposium is to develop a general approach for these two classes of product that will then
be reflected in monographs.
Learning objectives of this meeting are:
Gain a thorough insight and understanding of impurities control
Increase knowledge of the general monograph and chapter
Discover how manufacturers deal with and use these specifications
Learn more about the regulatory assessment of applications and assessors views and expectations
Hear case studies and examples of how these specifications are utilised in practice
Participants will have the possibility to meet and discuss with those involved in the elaboration of
monographs and to have an influence on the development of future specifications. Speakers will come from
industry, the European and American regulatory agencies, and the European Pharmacopoeia.
Who should attend?
This symposium should be attended by professionals from industry, in particular persons involved in the
manufacture and control of drug substances/products, including laboratory technicians, laboratory
consultants and those involved in the preparation of registration dossiers; from inspectorates, regulatory
agencies, OMCLs, and academic institutions.
Note: All participants from regulatory authorities and agencies will also be invited to attend a small meeting
to discuss the main outcomes of the symposium. More details will be provided following registration.
Register ONLINE to benefit from a reduced registration fee

Further information on the programme, accommodation and registration forms will be made available on the
EDQM internet site: http://www.pheur.org. How to contact us: Address: Public Relations Unit, EDQM, 226
avenue de Colmar BP 907, F-67029, Strasbourg, Cedex 1:
Tel: +33 (0) 3 88 41 30 30 (Dial 4 for Conferences);
Fax: + 33 (0) 3 88 41 27 71;
Internet: See under HELPDESK on our website: http://www.pheur.org/site/page_521.php
FOR MORE INFORMATION PLEASE VISIT THE WEBSITE: http://www.pheur.org
219
Location: Strasbourg, France
21-22 SEPTEMBER 2006
REGISTRATION FORM: SYMPOSIUM ON IMPURITIES CONTROL - ANTIBIOTICS AND PEPTIDES
_ __ __ _

650 *
or
300 *
REGISTRATION FEE ONLINE: If you register online, you benefit from a reduced registration fee of
600 * for industry, consultants, etc. and 250 * for national authorities, university, etc. (*See general conditions)
MEETING FOR NATIONAL AUTHORITIES DELEGATES ONLY
Yes, I shall be attending No, I shall not

First Name
Family Name
Company/Institution
Address for
Correspondence
Postcode
Town
Country
Telephone
Fax
E-mail
AREA OF ACTIVITY/
OCCUPATION:
University
Manufacturer of raw material

Manufacturer of medicines for human use for veterinary use
QA
QC
R&D
Regulatory affairs
Licensing
Pharmacopoeias
Inspection
OMCL
Other (please specify)__________________________________________________________________
PAYMENT (NEW)
2. BANK TRANSFER
3. CREDIT CARD
Company/Institution
Address
Postcode
Town
Country
Contact Name
Telephone
Fax
E-mail
Date
Signature
FOR MORE INFORMATION ABOUT CONFERENCES ORGANISED BY THE EDQM
220
IMPURITIES SYMPOSIUM: STRASBOURG

MONOPOLE METROPOLE HOTEL RESERVATION FORM

FAX: +33 (0) 3 88 32 82 55
To be filled in and sent by fax before the 30 August 2006

IMPURITIES SYMPOSIUM
Global reservation:
Contact reservations:
General information:
Cancellation policy:
Participant:
Hotel reservation:
BEST WESTERN
MONOPOLE
METROPOLE
50 single rooms have been reserved for Wednesday 20 and Thursday 21 September
2006. 10 single rooms have been reserved for Friday 22 and Saturday 23 September
2006.
BEST WESTERN - MONOPOLE METROPOLE HOTEL: 16 rue Kuhn,
67000 Strasbourg, France; Contact: Vronique Tel +33 (0)3 88 14 39 14,
Fax +33 (0) 3 88 32 82 55, E-mail: infos@bw-monopole.com.
The quotas of rooms reserved will be available until 30 August 2006.
After this date the availability and the negotiated prices will not be guaranteed. The
rooms should be reserved individually by each participant on a first come, first served
basis.
Before 30 August 2006 at no further cost, after this date and in case of no show one
night will be charged on your credit card.
Surname
Forename
Company/ employer
Address
City
Postal Code
Country
E-mail
Tel N
Fax N
Arrival day/hour /.....

Departure day/hour /..
Please indicate the selected nights by a circle
SINGLE
DBLE
NIGHT
20/09/2006
80 *
95 *
YES / NO
NIGHT
NIGHT
NIGHT
21/09/2006 22/09/2006 23/09/2006
YES / NO
YES / NO
YES / NO
* Taxes and breakfast inclusive
Method of payment:
Credit card number:
Credit card:
Visa
EuroCard / MasterCard Amex

Expiry date
Cardholder: ______________________________________________________
I authorise the BEST WESTERN MONOPOLE METROPOLE HOTEL to charge against my credit
card the amount equivalent to one night in order to block my reservation for the duration of the
seminar.
Date: _____________________
Signature: ________________________________________
HOTEL CONFIRMATION RESERVATION N:___________
Signature:________________
221
IMPURITIES SYMPOSIUM: STRASBOURG

MERCURE ST JEAN HOTEL RESERVATION FORM

FAX: +33 (0) 3 88 23 05 39
IMPURITIES SYMPOSIUM

Global reservation:
Participant:
Hotel reservation:
MERCURE
ST JEAN
20 single rooms have been reserved for Wednesday 20 September to Saturday 23

September 2006 (4 nights).
MERCURE ST JEAN HOTEL: 3 rue du Maire Kuss, 67000 Strasbourg, France
Contact: Tel +33 (0)3 88 32 80 80, Fax +33 (0)3 88 23 05 39,
E-mail: h1813@accor.com.
basis.
Surname
Forename
Company/ employer
Address
City
Postal Code
Country
E-mail
Tel N
Fax N

SINGLE
DBLE
NIGHT
20/09/2006
NIGHT
21/09/2006
NIGHT
22/09/2006
NIGHT
23/09/2006
92 *
102 *
YES / NO
YES / NO
YES / NO
YES / NO

Method of payment:
Credit card:
Visa
EuroCard / MasterCard
Credit card number:
Amex
Expiry date
Cardholder: ______________________________________________________
I authorise the MERCURE ST JEAN HOTEL to charge against my credit card the amount equivalent
to one night in order to block my reservation for the duration of the seminar.
Date: _____________________
Signature: ________________________________________
222
Signature:________________
NEW! 2006 SYMPOSIUM: STRASBOURG

NEW MICROBIOLOGY CHAPTERS OF THE
EUROPEAN PHARMACOPOEIA
2-3 October 2006
Duration: 2 days, Location: Strasbourg, France
***
FIRST ANNOUNCEMENT
The symposium will start with an overview of the Regulatory Environment for Medicines in Europe, with
the aim of understanding the roles of:
The National Pharmacopoeia Authorities
The European Agency for the Evaluation of Medicinal Products (EMEA)
The European Pharmacopoeia (PhEur)
The Pharmacopoeial Discussion Group (PDG), involving USP, JP and PhEur
The first day will be devoted to Microbial Contamination of Non-Sterile Products: the new, internationally
harmonised chapters of the European Pharmacopoeia (revised general chapters 2.6.12, 2.6.13 and 5.1.4).
Participants will learn about:
The principal changes described in the revised chapters: their impact on microbiological control
The implementation programme for the new chapters: indeed, a transition period is foreseen in
order for users to adapt to the revised chapters.
The second day will cover the new chapter on Alternative Methods for Control of Microbiological Quality
(general chapter 5.1.6). Participants will learn about:
The historical background of the chapter
The contents of the chapter
The regulatory acceptance of the new chapter
During both days, participants will have the opportunity to hear presentations and meet with the
specialists who have been involved in the elaboration of the new chapters. Representatives from national,
European and American regulatory authorities will also be present to facilitate the understanding of
marketing authorisation applications.
Who should attend?
This symposium should be attended by professionals from industry, in particular persons involved in the
manufacture and control of drug substances/products or the preparation of registration dossiers; from
inspectorates, regulatory agencies, OMCLs, and academic institutions.
Register ONLINE to benefit from a reduced registration fee

Further information on the symposium, accommodation and registration forms will be made available on
the EDQM internet site: http://www.pheur.org. How to contact us: Address: Public Relations Unit,
EDQM, 226 avenue de Colmar BP 907, F-67029, Strasbourg, Cedex 1:
Fax: + 33 (0) 3 88 41 27 71;
FOR MORE INFORMATION PLEASE VISIT THE WEBSITE: http://www.pheur.org
223
HELPDESK http://www.pheur.org/site/page_521.php: Go to the topic Conferences, Events, Public Relations and read
the question May I send my registration form via the EDQMs internet site for further instructions, or alternatively
register ONLINE, click on Events Register Online and select the name of the event.
_ __ __ _

650 *
or
300 *
600 * for industry, consultants, etc. and 250 * for national authorities, university, etc. (*See general conditions)
2-3 OCTOBER 2006
REGISTRATION FORM: SYMPOSIUM ON NEW MYCROBIOLOGY CHAPTERS OF THE PH.EUR

First Name
Family Name
Company/Institution
Address for
Correspondence
Postcode
Town
Country
Telephone
Fax
E-mail
AREA OF ACTIVITY/
OCCUPATION:
University
Manufacturer of raw material

Manufacturer of medicines for human use for veterinary use
QA
QC
R&D
Regulatory affairs
Licensing
Pharmacopoeias
Inspection
OMCL
Other (please specify)__________________________________________________________________
PAYMENT (NEW)
2. BANK TRANSFER
3. CREDIT CARD
Company/Institution
Address
Postcode
Town
Country
Contact Name
Telephone
Fax
E-mail
CANCELLATION CHARGES: I have read and accept the cancellation terms as stated on our web-site.
Date
Signature
FOR MORE INFORMATION ABOUT CONFERENCES ORGANISED BY THE EDQM
224
MICROBIOLOGY SYMPOSIUM: STRASBOURG

2-3 OCTOBER 2006
MONOPOLE METROPOLE HOTEL RESERVATION FORM

FAX: +33 (0) 3 88 32 82 55
Global reservation:
2-3 OCTOBER 2006
MICROBIOLOGY SYMPOSIUM
To be filled in and sent by fax before the 9 September 2006
Participant:
Hotel reservation:
BEST WESTERN
MONOPOLE
METROPOLE
10 single rooms have been reserved for Saturday 30 September 2006;

30 single rooms have been reserved for Sunday 1st and Monday 2 October 2006;
20 single rooms have been reserved for Tuesday 3 October 2006.
BEST WESTERN - MONOPOLE METROPOLE HOTEL: 16 rue Kuhn,
67000 Strasbourg, France; Contact: Vronique Tel +33 (0)3 88 14 39 14,
Fax +33 (0) 3 88 32 82 55, E-mail: infos@bw-monopole.com.
The quotas of rooms reserved will be available until 9 September 2006.
basis.
Before 9 September 2006 at no further cost, after this date and in case of no show one
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Address
City
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Country
E-mail
Tel N
Fax N

SINGLE
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80 *
95 *
YES / NO
NIGHT
NIGHT
NIGHT
01/10/2006 02/10/2006 03/10/2006
YES / NO
YES / NO
YES / NO
Method of payment:
Credit card:
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Amex
Expiry date
Cardholder: ______________________________________________________
I authorise the BEST WESTERN MONOPOLE METROPOLE HOTEL to charge against my credit
card the amount equivalent to one night in order to block my reservation for the duration of the
seminar.
Date: _____________________
Signature: ________________________________________
Signature:________________
225
2-3 October 2006, STRASBOURG
IBIS CENTRE GARE HOTEL RESERVATION FORM
FAX: +33 (0) 3 88 23 98 99
To be filled in and sent by fax before the 9 September 2006
2-3 October 2006 Location: Strasbourg, France
HOTEL RESERVATION FORM: MICROBIOLOGY SYMPOSIUM
Global reservation:
Participant:
Hotel reservation:
20 rooms have been reserved from Sunday 1st to Tuesday 3 October 2006
(3 nights)
IBIS CENTRE GARE HOTEL, 10 Place de la Gare, 67000 Strasbourg (France),
Contact: Tel +33 (0)3 88 23 98 98, Fax +33 (0)3 88 23 98 99,
E-mail: h3018@accor.com
The quotas of rooms reserved will be available until 9 September 2006.
After this date the availability and the negotiated prices will not be guaranteed.
The rooms should be reserved individually by each participant on a first come,
first served basis.
Before 20 September 2006 at no further cost, after this date and in case of no
show one night will be charged on your credit card.
Surname
Forename
Company/ employer
Address
City
Postal Code
Country
E-mail
Tel N
Fax N

NIGHT
NIGHT
NIGHT
01/10/2006 02/10/2006 3/10/2006
YES
YES
YES
62,- **
74,- **
74,- **
* Prices for a single or a double room are the same
** Taxes inclusive. Breakfast extra at 6,50,- per person/per day
Please tick the type of room you require Smoking room Non smoking room
IBIS
CENTRE
GARE
Method of payment:
SINGLE*
DOUBLE* TWIN*
Credit card: Diners
Visa
Credit card number:
EuroCard/MasterCard
Amex
Expiry date
Cardholder: ______________________________________________________
I authorise the IBIS STRASBOURG CENTRE GARE HOTEL to charge against my credit card the
amount equivalent to one night in order to block my reservation for the duration of the seminar.
Date: _____________________
Signature: ________________________________________
226
Signature:____________________
MICROBIOLOGY SYMPOSIUM: STRASBOURG

2-3 OCTOBER 2006
MERCURE ST JEAN HOTEL RESERVATION FORM

FAX: +33 (0) 3 88 23 05 39
Global reservation:
2-3 OCTOBER 2006
Participant:
Hotel reservation:
MERCURE
ST JEAN
20 single rooms have been reserved for Saturday 30 September, 30 single

rooms for Sunday 1st and Monday 2nd October 2007 and 20 single rooms for
Tuesday 3 October 2006.
MERCURE ST JEAN HOTEL: 3 rue du Maire Kuss, 67000 Strasbourg, France
Contact: Tel +33 (0)3 88 32 80 80, Fax +33 (0)3 88 23 05 39,
E-mail:h1813@accor.com
rooms should be reserved individually by each participant on a first come, first
served basis.
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Forename
Company/ employer
Address
City
Postal Code
Country
E-mail
Tel N
Fax N
Arrival day/hour /..... Departure day/hour /..

SINGLE
DBLE
NIGHT
30/09/2006
92 *
102 *
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NIGHT
NIGHT
NIGHT
01/10/2006 02/10/2006 03/10/2006
YES / NO
YES / NO
YES / NO

Method of payment:
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Credit card:
Visa
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Expiry date
Cardholder: ______________________________________________________
I authorise the MERCURE ST JEAN HOTEL to charge against my credit card the amount
equivalent to one night in order to block my reservation for the duration of the seminar.
Date: _______________
Signature: _____________________________________
HOTEL CONFIRMATION RESERVATION N:_______ Signature:________________
227
NEW! 2006 TRAINING COURSE: DUBLIN
THE EUROPEAN PHARMACOPOEIA 5TH EDITION

CHEMICALS
13-14 NOVEMBER 2006
Duration: 1.5 days, Location: Dublin, Ireland
***
FIRST ANNOUNCEMENT
The EDQM is delighted to announce its first training session in Dublin on the European Pharmacopoeia
on the 13-14 November 2006. The programme has been specifically designed to enable participants to
expand their knowledge and familiarise themselves with the work and procedures of the European
Pharmacopoeia. It will allow participants an opportunity to meet each other, exchange ideas and share
information.
The programme will cover:
An overview of the European regulatory system
Practical advice on how to use and interpret the European Pharmacopoeia, its monographs and
policies
An outline of how monographs are elaborated and revised, and how to participate in the process
The regulatory requirements of the Certification procedure, and offer guidance and advice on how
to prepare a successful dossier
The harmonisation programme, the relationship between the Pharmacopoeial Discussion Group
(PDG), Pharmacopoeias and ICH
During both days, participants will have the opportunity to hear presentations and meet with EDQM
scientific staff and it will also be possible to arrange one-to-one consultations. The services provided and
publications published by the EDQM will also be presented and information given on their use.
Who should attend?
This training session is intended for professionals from the pharmaceutical industry, regulatory agencies and
academic institutions.
Register online to benefit from a reduced registration fee

Further information:
To learn more about the programme, accommodation and to download the registration forms, please visit the
EDQM internet site: http://www.pheur.org. How to contact us: Address: Public Relations Unit, EDQM, 226
avenue de Colmar BP 907, F-67029, Strasbourg, Cedex 1:
Fax: + 33 (0) 3 88 41 27 71;
228
13-14 November 2006 Location: Dublin, Ireland
REGISTRATION FORM:
_ __ __ _

650 *
or
300 *
600 * for industry and 250 * for National Authorities, University. (*See general conditions)
ONE TO ONE MEETING
Yes, I am interested in reserving a meeting
No, I am not interested
Which topic(s)? EU Regulations Monographs, revisions Publications Certification
Internet

First Name
Family Name
Company/Institution
Address for
Correspondence
Postcode
Town
Country
Telephone
Fax
E-mail
AREA OF ACTIVITY/ Manufacturer of raw material
Manufacturer of medicines
OCCUPATION:
QA
QC
Licensing
Inspection
OMCL
Other (please specify)
for human use for veterinary use

R&D
Regulatory affairs
Pharmacopoeias
University
PAYMENT (NEW)
2. BANK TRANSFER
3. CREDIT CARD
Company/Institution
Address
Postcode
Town
Country
Contact Name
Telephone
Fax
E-mail
Date
Signature
FOR MORE INFORMATION PLEASE VISIT REGULARLY THE WEBSITE : http://www.pheur.org
229
ONE TO ONE CONSULTATION QUESTION FORM

13-14 November 2006 Location: Dublin, Ireland
ONE-TO-ONE RESERVATION FORM:
2006 TRAINING SESSION: DUBLIN

EUROPEAN PHARMACOPOEIA 5TH EDITION CHEMICALS
13-14 NOVEMBER 2006
If you would like to register for a one-to-one consultation (15 minutes) with a member of the EDQM
scientific team during the training course, please complete this consultation request form and send it
to the EDQM by fax to +33 3 88 41 27 71 or e-mail via the EDQM Helpdesk on the website:
Priority will be given to those who book in advance.
PARTICIPANT DETAILS
Title (Dr., Mr, Mrs, Ms)
First Name
Family Name
Company/Institution
E-mail
The time slots available for one-to-one consultations will be limited. The EDQM shall do all it can to
accommodate all interested participants.
If it is on a specific application, please mention the reference number
Your question:
Date
Signature
FOR MORE INFORMATION ABOUT TRAINING SESSIONS AND CONFERENCES ORGANISED BY THE EDQM
230
Certification of suitability
Certification of Suitability
of Monographs of the Ph. Eur.
LIST OF CERTIFICATES
CERTIFICATION SECTION OF THE EDQMS
WEBSITE
Visit the certication section of the EDQMs website
(www.pheur.org) and nd:
warnings/news;
answers to frequently asked questions;
general information on the certication procedure;
daily updated list of certicates granted.
LIST OF CERTIFICATES
Granted certicates
A daily updated list (more than 1900 certicates) is
available online in the certication section of our website
(www.pheur.org). The database can be searched by
substance name, certicate number or holder name. TSE
certicates can also be searched selectively. To print the
full list of certicates: select certicate number in the
database and search for CEP.
Voided or suspended certicates

A list of voided certicates is available online in the
certication section of our website (www.pheur.org) and
is also published in Pharmeuropa. This list includes:
the certicates not renewed after the 5-year validity
period (expired certicates);
the certicates withdrawn at the request of the
applicant (production stopped, site closed etc.);
the certicates suspended for a dened period or
withdrawn by the EDQM (for GMP deciencies or
insufcient information no longer in line with
regulatory requirements); these are identied by an
asterisk.
Warning: in the online search of the database, no
difference is made between suspended, expired or
withdrawn certicates.
Chemical purity and microbiological quality

These certicates have not been renewed / have been withdrawn.
Substance
Acetylcysteine
Certificate
R0-CEP 1997-071-Rev 00
Allopurinol
Alprazolam
Amiloride Hydrochloride
Amoxicillin Sodium
R0-CEP 1997-016-Rev 00
R0-CEP 1998-111-Rev 02
R0-CEP 1995-045-Rev 02
R1-CEP 1996-096-Rev 00
Amoxicillin Sodium
R1-CEP 1998-002-Rev 01
Amoxicillin trihydrate
Amoxicillin trihydrate; Compacted
Amoxicillin trihydrate
Amoxillin Sodium sterile
R0-CEP 1993-001-Rev 00
R0-CEP 1995-032-Rev 02
R0-CEP 1998-127-Rev 00
R1-CEP 1996-096-Rev 00
Ampicillin Sodium
Ampicillin Sodium; Sterile
Ampicillin Trihydrate
Ampicillin Trihydrate
Ampicillin, Anhydrous
Aspartic acid
Ascorbic acid
Ascorbic acid
R0-CEP 1995-004-Rev 01
R0-CEP 1998-132-Rev 00
R0-CEP 1994-010-Rev 00
R0-CEP 1992-001-Rev 02
R0-CEP 1994-009-Rev 00
R0-CEP 2000-295-Rev 02
R0-CEP 1995-019-Rev 01
R0-CEP 1997-035 Rev 02
Holder/Detenteur
Sterling SNIFF Italia; I 06073 Solomeo Di
Corciano (Perugia)
Siegfried Cms Ag/Ltd; CH 4800 Zofingen
Degussa AG; D 01445 Radebeul
Ribbon SRL Pharmaceutical and Chemical;
I 20145 Milano
Sandoz Industrial Products S.A.; E 08520
Barcelona
Flamma SpA; I 24125 Bergamo
Gist-Brocades BV; NL 2600 MA Delft
Smithkline Beecham; FR 35380 Pllan le Grand
Ribbon Srl Pharmaceutical and Chemical I-20145
Milano
Gist Brocades B.v; NL 2600 AK Delft
Gist-Brocades B V; NL 2600 MA Delft
Biochemie SA; E 08400 Granollers - Barcelona
Gist-Brocades B V; NL 2600 MA Delft
Bim Sifram Group; FR 75010 Paris
Pliva D D Zagreb; CRO 10000 Zagreb
Merck Kgaa D 64271 Darmstadt
* suspended, withdrawn or not renewed by EDQM following an inspection

** withdrawn by CEP Holder after suspension by EDQM as a result of an inspection
231
Atenolol
R0-CEP 1998-033 Rev 02
Beclometasone Dipropionate
Benzylpenicillin potassium
Benzylpenicillin potassium; sterile
Benzylpenicillin sodium; Sterile
Benzylpenicillin, benzathine;
Sterile, ( FA Process, sterile,
lecithin and polysorbate 80 coated;
Material Code Numbers 451028
and 451387)
(Soya lecithin coated 1,2 %)
Sterile, (Soya lecithin coated
1.2%)
Benzylpenicillin, procaine
Benzylpenicillin, procaine
Benzylpenicillin, procaine; (Soya
Lecithin Coated 1.2%), sterile
Benzylpenicillin, procaine; (Soya
lecithin coated)
Caffeine
R1-CEP 1992-013-Rev 00
R1-CEP 1992-003-Rev 00
R0-CEP 1996-086-Rev 01
R0-CEP 1996-085-Rev 01
R1-CEP 1994-018-Rev 04
Teva Pharmaceutical Industrie Ltd; IL 49131 Petah

Tiqva
Hoechst Marion Roussel; F 92800 Puteaux
Gist-Brocades Bv; NL 2600 MA Delft
Sandoz GmbH; A 6250 Kundl, Tyrol
R0-CEP 1995-013-Rev 01
R0-CEP 1996-087-Rev 01
R0-CEP 1995-012-Rev 02
R0-CEP 1996-011-Rev 02
R0-CEP 1996-037-Rev 02

Gist-Brocades Bv; NL 2600 MA Delft
Gist Brocades BV; S 645 41 Strngnas
R0-CEP 1996-010-Rev 01
R1-CEP 1997-047-Rev 00
Caffeine
Captopril
R0-CEP 2000-178-Rev 01
R0-CEP 1997-120-Rev 00
Carbamazepine
Carbamazepine
R0-CEP 1997-117-Rev 00
R0-CEP 1996-089-Rev 00
Carbasalate calcium
Cefadroxil monohydrate
R0-CEP 1997-056Rev 01
R0-CEP 1998-007 Rev 01
Cefadroxil monohydrate
R1-CEP 1998-019-Rev 00
Cefazolin sodium
Cefradine
R0-CEP 1998-054-Rev 03
R1-CEP 1997-106-Rev 00
Ceftriaxone Sodium; sterile
R0-CEP 1997-095-Rev 00
Cholecalciferol
Desmopressin
Diclofenac sodium
Diclofenac sodium
Dicloxacillin Sodium sterile
R0-CEP 1996-046-Rev 00
R0-CEP 1994-015-Rev 02
R0-CEP 1996-034-Rev 02
R0-CEP 1998-072-Rev 01
R0-CEP 1996-092- Rev 00
Digoxin
R0-CEP 1992-011-Rev 02
Hangzhou Minsheng Pharmaceutical Group Co;

RC 310 011 Hangzhou
KW Pfaffenschmidt GmbH; D 22459 Hamburg
Sinova Pharmaceuticals Co (Pte) Ltd; SGP 629534
Singapore
Vis Farmaceutici; I 35129 Padova
Fis-Fabbrica Sintetici SpA I-36041 Alte Di
Montecchio Maggi
DSM Minera BV, NL 3600 AC Maarssen
Dsm Anti-infectives Chemferm SA E 08130 Santa
Perpetua De Mogoda
Ranbaxy Laboratories Ltd; IND 110 019 New
Delhi
Amifarma S L; E 08389 Palafolls, Barcelona
Delhi
Delhi
Roche Vitamins Ltd; CH 4070 Basel
Ferring SA; F 94250 Gentilly
Degussa-Hls AG; D 01445 Radebeul
Klinge Pharma & Co Ltd; IRL County Kerry
Ribbon Srl Pharmaceutical and Chemical I-20145
Milano
Procter & Gamble Pharmaceuticals; F 92201
Neuilly Sur Seine Cdex
Dr Reddys Laboratories Ltd IND 500 016
Hyderabad
Ganes Chemicals Inc; USA NJ 08070 Pennsville
Wacker Chemie GmbH; D 81737 Munchen
Dow Corning France; F 69432 Lyon Cedex 03
Kraemer And Martin Pharma Handels Gmbh; D
47804 Krefeld
Eli Lilly SA; IRL County Cork
Diltiazem Hydrochloride Process I R0-CEP 1997-081 Rev 01

Dimenhydrinate
Dimeticone
Dimeticone; (20, 50)
Doxycycline hyclate
R0-CEP 1998-081-Rev 00
R0-CEP 1998-016-Rev 00
R0-CEP 1995-048-Rev 02
R0-CEP 1996-063-Rev 00
Fluoxetine hydrochloride
R0-CEP 1999-046-Rev 02

232
Flucloxacillin Sodium; n-butyl

acetate process; powder;
compacted
Folic acid
Fructose
Gelatin limed hide gelatin
Guaifenesin
Haloperidol
Hydrocortisone
Hydrocortisone Acetate
Ibuprofen
Iopamidol
Isosorbide Monohydrate, diluted
70% and 80%
Isosorbide Mononitrate, Diluted
Isoleucine
Itraconazole
Ketoconazole
Ketoprofen
Ketoprofen
Levamisole Hydrochloride
Loperamide Hydrochloride
Mebendazole; Polymorph C
Metamizole sodium
Methotrexate
Metronidazole
Miconazole nitrate
Midazolam
Minocycline Hydrochloride
Nimodipine
Paracetamol
Paracetamol
Paracetamol
Paracetamol
Pentoxifylline
Phenazone
Phenoxymethylpenicillin
potassium
Phenylephrine Hydrochloride
Potassium clavulanate
R0-CEP 1996-029-Rev 04
Biochemie SA; E 08520 Les Franqueses Del

Valls
R0-CEP 1994-002-Rev 00
R0-CEP 1997-013-Rev 00
R0-CEP 2000-386-Rev 00
BASF Aktiengesellschaft; D 67056 Ludwigshafen

Danisco Sweeteners Ltd; GB RH1 6YS Redhill
Gelita Group (DGF Stoess, Kind & Knox), D
69412 Eberbach
R0-CEP 2002-194-Rev 00*
Schtz & Co (GmbH & Co); D 20095 Hamburg
R0-CEP 2000-179-Rev 01
Janssen Pharmaceutical Ltd IRL County Cork
R0-CEP 1996-043-Rev 00
Diosynth BV; NL 5340 BH Oss
R0-CEP 1995-042-Rev 00
Diosynth BV; NL 5340 BH Oss
R0-CEP 1997-124- Rev 02
BASF PLC ; GB NE23 9JL Cramlington
R0-CEP-1999-130 Rev 02
Recordati SpA; I 20148 Milano
R0-CEP 2001-329-Rev 00 ** Calao SRL, I 20151 Milano
R0-CEP 1999-107-Rev 00
R0-CEP 1998-041-Rev 03
R0-CEP 2001-150-Rev 01
R0-CEP 1997-105-Rev 02
R0-CEP 1994-019-Rev 01
R0-CEP 1995-046-Rev 02
R1-CEP 1993-006-Rev 00
R1-CEP 1996-088-Rev 01
R0-CEP 1997-036-Rev 00
R0-CEP 2001-220-Rev 00 *
Whyte Chemicals; GB N3 2UA Finchley, London

Amino GmbH; D 38373 Frellstedt
Janssen Pharmaceutical Ltd; IRL County Cork
Aventis Pharma Ltd; GB RM10 7XS Dagenham
Nordic Synthesis Ab; S 69185 Karlskoga
Janssen Pharmaceutica NV; B 2340 Beerse
Janssen Pharmaceutica NV; B 2340 Beerse
Vani Chemicals & Intermediates Ltd ;
IND 500 055 Hyderabad
R0-CEP 1996-026-Rev 00
Pfizer And Pfizer Mack; D 89252 Illertissen
R0-CEP 1993-004-Rev 00
Rhne-Poulenc Rorer SA; F 92165 Antony Cedex
R0-CEP 1998-153-Rev 01
Janssen Pharmaceutical Ltd IRL County Cork
R0-CEP 1998-114-Rev 01
Diosynth BV; NL 5349 AB Oss
R0-CEP-1999-072 Rev 02
Nippon Kayaku Co. Ltd; J 102-8172 Tokyo
R0-CEP-1999-105 Rev 02
Siegfried Ltd; CH 4800 Zofingen
R0-CEP 2000-002-Rev 00 ** Indukern Chemie AG; CH 8952 Schlieren
R0-CEP 1996-057-Rev 01
BASF Corporation; USA 78343 Bishop, Texas
R0-CEP 2002-020-Rev 01*
Farmson Analgesics (Unit of Farmson
Pharmaceutical Gujarat Pvt Ltd); IND 391 340
Nandesari, Dist Vadodara, Gujarat
R0-CEP 2001-092-Rev 02
Bristol Laboratories Ltd; GB HA1 2EN Middlesex
R0-CEP 1996-035-Rev 01
Degussa-Hls AG; D 01445 Radebeul
R0-CEP 2001-042-Rev 00 * Vani Chemicals & Intermediates Ltd ;
R1-CEP-1994-017 Rev 02
DSM Anti-Infectives; NL 2613 AX Delft
R0-CEP 1995-044-Rev 02
R0-CEP 1999-112-Rev 00
Potassium clavulanate; (The

Netherlands)
Propyphenazone
R0-CEP 1996-012-Rev 02
Pyridoxine Hydrochloride
Quinidine sulphate
Quinine hydrochloride
Quinine hydrochloride
Quinine sulphate
Quinine sulphate
Selegiline Hydrochloride
R1-CEP 1994-003-Rev 02
R1-CEP 1992-007-Rev 00
R0-CEP 1997-030-Rev 01
R1-CEP 1992-009 Rev 03
R0-CEP 1998-003-Rev 01
R1-CEP 1992-006- Rev 02
R0-CEP 1997-020-Rev 00
R0-CEP 2001-041-Rev 00 *

Smithkline Beecham Pharmaceuticals; GB BN14
8QH Worthing
Vani Chemicals & Intermediates Ltd ;
BASG Aktiengesellschaft; D 67056 Ludwigshafen
Roche Diagnostics GmbH; D 68298 Mannheim
Roche Diagnostics GmbH; D 68298 Mannheim
Teva Group Bulk Pharmaceuticals Division; IL
49131 Petah Tiqva

233
Sodium Valproate
Sulfadiazine
Sulfadiazine
R0-CEP 1996-015-Rev 02
R0-CEP 1992-008-Rev 00
R0-CEP 1995-040-Rev 01
Sulfamethoxazole
Tamoxifen Citrate
Terfenadine
Terfenadine
Theophylline
Thiamine Hydrochloride
Thiamine nitrate
Trimethoprim
Vinblastine Sulphate
R0-CEP 1995-026-Rev 02
R0-CEP 1997-024-Rev 01
R0-CEP 1997-003-Rev 00
R0-CEP 1995-037-Rev 01
R1-CEP 1998-018-Rev 00 *
R1-CEP 1994-005-Rev 02
R1-CEP 1994-006-Rev 02
R0-CEP 1999-132-Rev 02
R1-CEP 1992-005-Rev 00

DSM Minera BV; NL 3600 AC Maarssen
Southwest Synthetic Pharmaceutical General
Factory; RC 400 025 Chongqing
Shionogi & Co Ltd; J 541-0045 Osaka
Ranbaxy Laboratories Ltd; IND 144 533 Toansa
Avik Pharmaceutical Ltd; IND 396 195 Vapi
KW Pfannenschmidt GmbH; D 22459 Hamburg
Welding GmbH & Co KG; D 20354 Hamburg
Eli Lilly SA - Irish Branch; IRL County Cork

*** CEP restored
Transmissible spongiform encephalopathy risk products

The following certicates have been withdrawn.
Substance
Bordet Gengou Agar Bae
(Product codes: 211050 and
211051)
Brain Heart Infusion
Chocolate Agar Plates
EMB Agar, Levine EMB Agar
(Product codes 276100, 276200,
276300, 276400, 251000, 212088,
212095 and 254000)
Gelatin, Alkaline-Processed
Chromium-Tanned Hide GelatinSPA
Gelatin, Acid bone gelatin
Certificate
R0-CEP 2000-318-Rev 02
Holder/Detenteur
BD Diagnostic Systems ; USA 21152 Sparks
R0-CEP 2000-267-Rev 00
R0-CEP 2001-328-Rev 01
R0-CEP 2000-315-Rev 01

R0-CEP 2000-113-Rev 02
Croda Colloids Ltd; GB WA8 8UB Widnes,

Cheshire
R0-CEP 2000-227-Rev 03
Gelatin, Limed bone gelatin
R0-CEP 2000-228-Rev 03
Gelatin, Limed bone gelatin

Gelatin limed hide gelatin
R0-CEP 2000-085-Rev 05
R0-CEP 2000-386-Rev 00
Gelatin US Enzymatic Bone

Gelatin Hydrolysate
Gelatin acid bone gelatin,
American origin
American origin
including NaOH treatment
Gelatin; / Acid bone gelatin;
(French [European] origin)
PPLO Broth w/o CV, FD
R0-CEP 2002-153-Rev 01

Cheshire
Cheshire
Nitta Gelatin Inc; J 556-0022 Osaka
Gelita Group (DGF Stoess, Kind & Knox), D69412 Eberbach
Gelita Group; D 69412 Eberbach
R0-CEP 2000-139-Rev 05
PB Gelatins; B 1800 Vilvoorde
R0-CEP-2000-026-Rev 03
Rousselot SAS; F 92641 Boulogne Billancourt

Cedex
Rousselot SAS; F 92641 Boulogne Billancourt
Cedex
Skw Biosystems; F 92641 Boulogne Billancourt
Cedex
234
R0-CEP-2001-411-Rev 02
R0-CEP 2000-030-Rev 00
R0-CEP 2002-084-Rev 01
Readers Tribune
Readers Tribune
The Secretariat of the European Pharmacopoeia recalls that articles reect the authors opinions. In order that all
opinions are taken into account, concerned users and readers of Pharmeuropa are invited to send their comments
to the Secretariat. Readers are reminded that details regarding contributions made to Pharmeuropa are cited on the
rear cover of this issue.
HERBAL REFERENCE STANDARDS

Dr K. Helliwell
The current evaluation and gradual introduction into
the European Pharmacopoeia (Ph. Eur.) of Chemical
Reference Standards for use in Herbal Drug and Herbal
Drug Preparation monographs appears to be a suitable
point to initiate a discussion as to the types of reference
standards that are applicable to such monographs.
However, before embarking upon this discussion, it is
necessary to understand the signicant changes that
have taken place within the herbal medicinal products
sector of the pharmaceutical industry during the last
few decades. Within this context, it is worth noting that,
30-40 years ago:
a signicant decrease in the training and expertise

in the traditional physical methods for the
assessment of herbal drugs, particularly microscopical
identication.
a signicant increase in regulatory requirements;
herbal drug preparation reference standards;
the introduction of clinical trial based herbal

medicinal products;
chemical reference standards.
the introduction of new and improved analytical

techniques;
HERBAL DRUG REFERENCE STANDARDS
As a result, during this period, the emphasis for quality

control has changed from a physical to a chemical
evaluation. Yet, it is only within the last 12-18 months
that Chemical Reference Standards specically for use
in the application of Herbal Drug and Herbal Drug
Preparations monographs are being evaluated and
gradually introduced into the Ph. Eur. (e.g., Boldine CRS,
Coumarin CRS and Ruscogenins CRS). Table 1 lists the
there was a limited range of major pharmacopoeial
number of herbal drug (excluding exudates) and herbal
herbal drugs available, these included: solanaceous
drug preparation monographs that have been included in
plants (belladonna, hyoscyamus and stramonium),
various editions of the Ph. Eur. since 1971. This clearly
anthraquinone containing plants (aloes, cascara,
shows that a signicant increase in the inclusion of such
frangula, senna leaves and pods), ipecacuanha and
monographs has taken place within the last decade. This
opium;
trend is set to continue with the intention of establishing
quantitative determinations, if applicable, were mainly monographs on traditional Chinese herbal drugs (and
herbal drug preparations) in the Ph. Eur. (Pharmeuropa
titrametric or spectrophotometric;
17.4). It is also worth noting that herbal medicinal
many other plants used as herbal drugs had no assay
products from other cultures, for example, Ayurvedic
or equivalent; quality control was based primarily
herbal drugs, are to be included in some National
on macroscopy, microscopy, extractable matter, ash
Pharmacopoeias, for example, the British Pharmacopoeia.
values, foreign matter, loss on drying etc.;
In consideration of the changes that have taken place,
chromatography as a routine quality control tool,
this appears to be an appropriate time to examine the
either qualitative or quantitative, was in its infancy;
types of reference standards that could, or should, be
made available for the evaluation of herbal drugs/herbal
analysts were appropriately trained and familiar with
the physical rather than chemical characteristics of drug preparations/herbal medicinal products. In this
context, the following types of reference standards are
the herbal drugs which they were handling;
proposed:
and that over the intervening period, the changes have
herbal drug reference standards;
been dramatic:
a signicant increase in the overall use of herbal

medicinal products, both Western herbal medicines
and those from other cultures (traditional Chinese,
Ayurvedic, etc.);
a perceived necessity to have a qualitative and
quantitative chemical evaluation of herbal drugs,
herbal drug preparations and herbal medicinal
products;
Diagram 1 represents the inter-relationship between

herbal drugs, herbal drug preparations and herbal
medicinal products. This demonstrates that not only
highly processed herbal drug preparations, such as
extraction products (e.g., dry extracts and tinctures), may
be incorporated into herbal medicinal products, but that
a herbal drug, in a cut (e.g., herbal tea) or powdered state
(e.g., tablet/capsule), may also form the basis of a herbal
medicinal product.
Dr K. Helliwell. William Ransom & Son plc, Hitchin, Herts, SG5 1RT, England
235
Readers Tribune
As has already been stated, expertise in the eld of the

microscopical identication of herbal drugs is on a steep
decline. However,
microscopical identication is still a mandatory
element of pharmacopoeial monographs for herbal
drugs;
more and more monographs for herbal drugs are
being introduced into pharmacopoeias;
many herbal drugs in the market place are traded in
a cut or powdered form.
One proposal to attempt to ease the present situation is to
incorporate into the Ph. Eur. diagramatic representations
of the diagnostic microscopical characters of herbal
drugs (Pharmeuropa 17.2 and 17.3). This may have
both positive and negative consequences, the negative
being that these diagramatic representations become
no more than a tick-list for analysts undertaking the
microscopical identication and that quite obvious
characters from other sources, such as adulterants, may
be ignored. It should be remembered that diagnostic
characters are characteristic of but not necessarily
exclusive to a particular herbal drug.
An alternative approach would be to make available
Herbal Drug Reference Standards (HDRS) of
authenticated herbal drugs (freed from foreign matter) in
the same way that Chemical Reference Standards (CRS)
are available. The following points should be noted with
respect to the use of such standards:
HDRS for a specic herbal drug would be supplied
milled to the particle size stated for the microscopical
identication in the monograph for that herbal
drug and would be designated as a primary standard
(HDRS1);
have been permitted in addition to G.glabra L. The

description of the microscopical diagnostic characters
has not changed. It would be extremely useful to
have HDRS for each of these species, particularly as
the content of the measured constituent (glycyrrhizic
acid) varies signicantly between the species;
Pharmacopoeial Expert Groups/Committees are
looking at the elaboration of pharmacopoeial
monographs on traditional Chinese herbal drugs:
in traditional Chinese medicine substitution of one
herbal drug for another is not uncommon and the
same Chinese name can apply to more than one
herbal drug;
HDRS will be essential for these, often unfamiliar,
materials.
HERBAL DRUG PREPARATION REFERENCE
STANDARDS AND CHEMICAL REFERENCE
STANDARDS
Herbal drug preparation reference standards and
chemical reference standards are chemical standards to
be used for the qualitative and quantitative evaluation
of chemical constituents which are either characteristic
of, or should be restricted in, or absent from a specied
herbal drug, herbal drug preparation or herbal medicinal
product.
The following should be noted in the use of these two
types of chemical standards.
Quantitative aspects
the primary function of a HDRS would be to allow

conrmation of the microscopical identity of a herbal
drug by comparison against a reference standard;
Assay: constituents to be quantied may be either

active components responsible for the therapeutic
activity of the herbal drug/herbal drug preparation/
herbal medicinal product, or, where the active
components are not as yet identied, may be
marker compounds characteristic of the herbal
drug/herbal drug preparation/herbal medicinal
product which may or may not be chemically
similar to any active component.
it is preferable to compare against authentic material

rather than a diagramatic representation as other
characters besides diagnostic characters should be
assessed;
The constituent(s) to be quantied usually from

a small percentage of the overall composition of
the herbal drug/herbal drug preparation/herbal
medicinal product; often, less than 1 per cent.
HDRS would aid the identication of the presence

of any foreign elements, not present in the HDRS,
should adulterants/foreign matter be present in the
test sample;
In many cases there is no economically viable

source of the pure individual chemical
constituent(s) to be quantied.
microscopical characters of dried herbal drugs do not

change with time;
HDRS would be useful in identifying the presence

of a stated herbal drug in herbal drug preparations
(e.g., cut herbal drugs, powdered herbal drugs) and in
herbal medicinal products (e.g., herbal teas, tablets,
capsules).
Such HDRS would be physical standards to be used
for the microscopical identication of those structural
components which are characteristic of a specied herbal
drug in order to conrm the identity of that herbal drug
or indicate its presence in a herbal drug preparation or a
herbal medicinal product.
Examples of applications for HDRS would be:
liquorice root: recently two further species,
Glycyrrhiza inata Bat. and G.uralensis Risch
236
In some cases quantication is based upon a series

of related chemical constituents rather than a single
compound.
It is preferable to have the same constituent(s)
quantied at each stage of the cascade:
Herbal Drug
r
Herbal Drug Preparation
r
Herbal Medicinal Product (see Diagram1)
Qualitative aspects
These may be critical as it may be a specic constituent
which allows differentiation between plant species, some
Readers Tribune
of which may be toxic e.g., star anise (Illicium verum)

and poisonous Illicium species.
HERBAL DRUG PREPARATION REFERENCE
STANDARDS
An alternative designation, for example, HDPRS1, would

seem more appropriate for these primary standards as
they do not meet the strict criteria required of a CRS1.
CHEMICAL REFERENCE STANDARDS
These are herbal drug preparations derived from the

Key considerations
processing of herbal drugs, usually extraction products
(e.g., dry extracts or tinctures) whose relevant properties
Where possible the chemical reference standard
have been determined and which are considered to be
should meet all of the criteria for chemical
appropriate for use as a primary standard for the intended
reference standards for synthetic drugs (as
purpose.
proposed in the draft Ph. Eur. document
5.12. Pharmaceutical reference standards
Key considerations
PA/PH/SG (04) 45 ANP).
Is a chemical reference standard an option (this
should always be the preferred route, if viable)?
Where this is not possible, particularly with regard
to conrmation of assigned content via a mass
Has the specied constituent(s) for which the
balance, it would still be preferable to have a
herbal drug preparation is to be used as a standard
designated primary standard even if the content of
sufcient stability to make it a useful reference
specied constituent(s) had an assigned value as low
standard (particularly important when the herbal
as 80 per cent m/m.
drug preparation is water/ethanol based, such as a
tincture)?
The content of specied constituent(s) for which
the herbal drug preparation is to be used as a
standard is not an absolute value, but an assigned
value determined by a specied method.
Currently all primary Chemical Reference Standards are

designated CRS1, perhaps chemical reference standards
for herbal products which do not meet all of the CRS1
criteria should have a different designation, for example,
Chemical Reference Standard (Herbal Products) CRS1 (HP).
It is often not possible to verify the assigned content

of the specied constituent(s) by a calculation of the At present, there are few primary reference standards
mass balance.
for herbal drugs/herbal drug preparations/herbal
medicinal products. Those that are in the process of
Depending upon the herbal drug and the type of
herbal drug preparation, the assigned content could being introduced are chemical standards. This early
stage in the introduction of reference standards for
be as high as 40-50 per cent m/m for some dry
herbal products is an ideal opportunity for industry/
extracts and signicantly less than 1 per cent m/m
pharmacopoeial authorities/regulatory authorities to
for some tinctures.
dene the types of reference standard that would best
Currently, it is proposed that Herbal Drug Preparation
satisfy the needs of this sector of the pharmaceutical
Reference Standards are designated as CRS1.
industry and agree a nomenclature for such standards
that gives clarity and transparency in their use.
Table 1 - Herbal drug and herbal drug preparation monographs included in

various editions of the European Pharmacopoeia
Year
Number of Herbal Drugs
Number of Herbal Drug

Preparations
1st Edition Volume II
1971
12
2nd Edition 17th Fascicle
1993
31
5*
3rd Edition
1997
37
4th Edition
2002
96
13
5th Edition up to and

including Supplement 5.4
2006
119
27
* General monographs on Extracts and Tinctures (1992) had recently been introduced
237
Readers Tribune
Diagram 1 - Inter-relationships of herbal drugs/herbal drug preparations/herbal medicinal products

Herbal drug
Cut, Powdered
Extracted (dry extracts, tinctures, etc.)
Herbal drug preparation
238
Solid dosage forms (tablets, capsules)

Oral liquid products (tinctures, mixtures, etc.)
Semi-solid dosage forms (ointments, creams)
Herbal medicinal product
Teas
Solid dosage forms (tablets, capsules)
Adenosine
Draft monographs and

general texts for comment
IMPORTANT NOTICE
This section contains proposals for new and revised monographs and general texts, intended for inclusion in the
European Pharmacopoeia and submitted for public comment. You can comment through the appropriate national
authority at the address listed on the inside back cover of this issue. Readers from countries that are not members of the
European Pharmacopoeia Commission should send their comments directly to the European Directorate for the Quality
of Medicines. In order to facilitate the work of the Secretariats of the national authorities and the European Directorate
for the Quality of Medicines who collect the comments, please mention in any correspondence the PA/PH reference
number indicated at the beginning of each text. Comments that propose modifications of limits should be supported by
analytical data obtained on a significant number of batches. Proposed changes of methodology should be supported by
experimental results of a comparative trial of the method published in Pharmeuropa for comment and the proposed
alternative. Only comments sent before 30 June 2006 will be considered before the final version is prepared.
It is stressed that these proposals have not been adopted by the European Pharmacopoeia Commission and must not
be regarded as official standards.
In the case of proposals for revision, text to be deleted is crossed out and replacements or additions are underlined.
In certain cases, draft monographs require, for appropriate checking, the use of a reference material that is not yet
commercially available; in exceptional circumstances, we will try to make the necessary substance available; please
enquire to the European Directorate for the Quality of Medicines.
Reference: PA/PH/Exp. 10D/T (05) 35 ANP
TESTS
Solution S. Suspend 5.0 g in 100 ml of distilled water R and
NOTE ON THE MONOGRAPH
heat to boiling. Allow to cool, filter with the aid of vacuum
It is proposed to replace the TLC test for related substances and dilute to 100 ml with distilled water R.
with an LC test. Only comments on this test are requested. Appearance of solution. Solution S is colourless (2.2.2,
XXXX:1486 Method II).
Acidity or alkalinity. To 10 ml of solution S, add 0.1 ml
of bromocresol purple solution R and 0.1 ml of 0.01 M
ADENOSINE
hydrochloric acid. The solution is yellow. Add 0.4 ml of
0.01 M sodium hydroxide. The solution is violet-blue.
Adenosinum
Specific optical rotation (2.2.7) : 45 to 49 (dried
substance).
Dissolve 1.25 g in 1 M hydrochloric acid and dilute to
50.0 ml with the same acid. The specific optical rotation is
determined within 10 min of preparing the solution.
Related substances. Examine by thin-layer chromatography
(2.2.27), using a TLC silica gel F254 plate R.
Test solution. Dissolve 0.20 g of the substance to be
examined in dilute acetic acid R with slight heating and
dilute to 5 ml with the same acid.
Reference solution (a). Dilute 1 ml of the test solution to
C10H13N5O4
Mr 267.2 100 ml with water R.
Reference solution (b). Dissolve 10 mg of adenosine CRS
DEFINITION
and 10 mg of adenine CRS in dilute acetic acid R, with
9--D-Ribofuranosyl-9H-purin-6-amine.
heating if necessary, and dilute to 10 ml with the same acid.
Apply to the plate 5 l of each solution. Develop over a
Content : 99.0 per cent to 101.0 per cent (dried substance).
path of 12 cm using a mixture of 10 volumes of water R,
30 volumes of concentrated ammonia R and 60 volumes
CHARACTERS
of propanol R. Allow the plate to dry in a current of warm
Appearance : white or almost white, crystalline powder.
air and examine in ultraviolet light at 254 nm. Any spot in
Solubility : slightly soluble in water, soluble in hot water,
the chromatogram obtained with the test solution, apart
practically insoluble in ethanol (96 per cent) and in
from the principal spot, is not more intense than the spot
methylene chloride. It dissolves in dilute mineral acids.
in the chromatogram obtained with reference solution (a)
(1 per cent). Spray with a 5 g/l solution of potassium
mp : about 234 C.
permanganate R in 1 M sodium hydroxide. Allow the plate
to dry in a current of warm air and examine in daylight. Any
IDENTIFICATION
spot in the chromatogram obtained with the test solution,
Infrared absorption spectrophotometry (2.2.24).
apart from the principal spot, is not more intense than
the spot in the chromatogram obtained with reference
Comparison : adenosine CRS.
239
Adenosine
solution (a) (1 per cent). The test is not valid unless the
chromatogram obtained with reference solution (b) shows
two clearly separated spots.
Liquid chromatography (2.2.29).
System suitability : reference solution (b) :

resolution : minimum 1.5 between the peaks due to
impurities A and G.
Limits :
Solvent mixture. Dissolve 6.8 g of potassium hydrogen
correction factors : for the calculation of content,
sulphate R and 3.4 g of tetrabutylammonium hydrogen
multiply the peak areas of the following impurities by
sulphate R1 in water R and dilute to 1000 ml with the
the corresponding correction factor : impurity A = 0.6 ;
same solvent. Adjust to pH 6.5 with a 120 g/l solution of
impurity G = 1.4 ;
potassium hydroxide R.
impurity A : not more than twice the area of the principal
Test solution. Dissolve 20 mg of the substance to be
peak in the chromatogram obtained with reference
examined in the mobile phase and dilute to 20.0 ml with the
solution (a) (0.2 per cent) ;
mobile phase.
impurity G : not more than the area of the principal peak
Reference solution (a). Dilute 1.0 ml of the test solution
(0.1 per cent) ;
to 100.0 ml with the mobile phase. Dilute 1.0 ml of this
solution to 10.0 ml with the mobile phase.
unspecified impurities : for each impurity, not more
than the area of the principal peak in the chromatogram
Reference solution (b). Dissolve 5 mg of adenine CRS
obtained with reference solution (a) (0.10 per cent) ;
(impurity A) and 5 mg of inosine R (impurity G) in the mobile
total : not more than 5 times the area of the principal peak
phase and dilute to 50 ml with the mobile phase. Dilute
4.0 ml of this solution to 100.0 ml with the mobile phase.
(0.5 per cent) ;
Column :
disregard limit : 0.5 times the area of the principal peak
size : l = 0.25 m, = 4.6 mm ;
(0.05 per cent).
stationary phase : end-capped octadecylsilyl silica gel
for chromatography R (5 m)(1) ;
Chlorides (2.4.4) : maximum 100 ppm.
temperature : below 30 C.
Dilute 10 ml of solution S to 15 ml with water R.
Sulphates (2.4.13) : maximum 200 ppm, determined on
Mobile phase : mix 40 volumes of a 0.1 g/l solution of
solution S.
sodium azide R and 60 volumes of the solvent mixture.
Ammonium (2.4.1, Method B) : maximum 10 ppm,
Flow rate : 1.5 ml/min.
determined on 0.5 g.
Detection : spectrophotometer at 254 nm.
Prepare the standard using 5 ml of ammonium standard
Injection : 20 l.
solution (1 ppm NH4) R.
Run time : 1.5 times the retention time of adenosine.
Loss on drying (2.2.32) : maximum 0.5 per cent, determined
on 1.000 g by drying in an oven at 100-105 C.
Relative retention with reference to adenosine (retention
Sulphated ash (2.4.14) : maximum 0.1 per cent, determined
time = about 13 min) : impurity A = about 0.3 ;
on 1.0 g.
impurity G = about 0.4.
The following chromatogram is shown for information but will not be published in the European Pharmacopoeia.
1. impurity F
2. impurity A
3. impurity G
4. impurity H
5. adenosine
Figure 1486.-1. Chromatogram for the test for related substances of adenosine : solution of adenosine spiked with
impurities A, F, G and H
(1)
Hypersil ODS, Symmetry RP18 and Nucleodur C18 Pyramid are suitable.
240
ASSAY
Dissolve 0.200 g, warming slightly if necessary, in a mixture
of 20 ml of acetic anhydride R and 30 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 26.72 mg
of C10H13N5O4.
IMPURITIES
Specified impurities : A, G.
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 5.10. Control of impurities in substances for
pharmaceutical use) : B, C, D, E, F, H.
H. 2-amino-9--D-ribofuranosyl-1,9-dihydro-6H-purin-6-one
(guanosine).
Reagents
Inosine. C10H12N4O5. (Mr 268.23). XXXXXXX. [58-63-9].
Hypoxanthine 9--D-ribofuranoside.
mp : 222 C to 226 C.
A. adenine,
Reference: PA/PH/Exp. 10B/T (05) 96 ANP
B. D-ribose,

It is proposed that the monograph be revised, in particular
to bring the related substances test into line with the
current policy for the control of impurities.
XXXX:1287
ALFUZOSIN HYDROCHLORIDE
Alfuzosini hydrochloridum
C. R = H : adenosine 3-(dihydrogen phosphate),

D. R = PO3H2 : adenosine 3-(trihydrogen diphosphate),
E. R = PO2H-O-PO3H2 : adenosine 3-(tetrahydrogen
triphosphate),
C19H28ClN5O4
Mr 425.9
DEFINITION
(RS)-N-[3-[(4-Amino-6,7-dimethoxyquinazolin-2yl)(methyl)amino]propyl]tetrahydrofuran-2-carboxamide
hydrochloride.
Content : 98.5 99.0 per cent to 101.0 per cent (anhydrous
substance).
CHARACTERS
Appearance : white or almost white, crystalline powder,
slightly hygroscopic.
Solubility : freely soluble in water, sparingly soluble in
ethanol (96 per cent), practically insoluble in methylene
chloride.
F. 1--D-ribofuranosylpyrimidine-2,4(1H,3H)-dione (uridine),
G. 9--D-ribofuranosyl-1,9-dihydro-6H-purin-6-one (inosine),
IDENTIFICATION
A. Infrared absorption spectrophotometry (2.2.24).
Examine the substances prepared as discs.
Comparison : alfuzosin hydrochloride CRS.
B. To 1 ml of solution S (see Tests) add 1 ml of water R. The
solution It gives reaction (a) of chlorides (2.3.1).
TESTS
Solution S. Dissolve 0.500 g in carbon dioxide-free water R
and dilute to 25.0 ml with the same solvent.
pH (2.2.3) : 4.0 to 6.0 5.5 for freshly prepared solution S.
241
Optical rotation (2.2.7) : 0.10 to + 0.10, determined on

solution S.
Related substances. Liquid chromatography (2.2.29).
Test solution. Dissolve 20.0 40.0 mg of the substance to be
examined in the mobile phase and dilute to 100.0 ml with
the mobile phase.
Reference solution (a). Dilute 1.0 ml of the test solution to
50.0 100.0 ml with the mobile phase. Dilute 5.0 1.0 ml of
this solution to 20.0 10.0 ml with the mobile phase.
Reference solution (b). Dissolve 5 mg of alfuzosin
impurity A CRS in the mobile phase and dilute to 25 ml
with the mobile phase. To 1 ml of the solution, add 1 ml
of the test solution and dilute to 100 ml with the mobile
phase. Dissolve 4 mg of alfuzosin for system suitability CRS
(containing impurities A and D) in the mobile phase and
dilute to 10.0 ml with the mobile phase.
The chromatographic procedure may be carried out using :
a stainless steel column 0.15 m long and 4.6 mm
in internal diameter packed with microparticulate
octadecylsilyl silica gel for chromatography R (5 m),
with a carbon loading of 18.5 per cent, a specific surface
area of 320 m2/g, a pore size of 15 nm, and end-capped
with hexamethyldisilane.
Column :
size : l = 0.15 m, = 4.6 mm ;
stationary phase : octadecylsilyl silica gel for
chromatography R (5 m)(2).
Mobile phase : mix 1 volume of tetrahydrofuran R,
20 volumes of acetonitrile R and 80 volumes of a solution
prepared as follows : dilute 5.0 ml of perchloric acid R in
900 ml of water R, adjust to pH 3.5 with dilute sodium
hydroxide solution R and dilute to 1000 ml with water R.
Inject 20 l of reference solution (b). Adjust the sensitivity
of the system so that the height of the two peaks in the
chromatogram obtained is at least 50 per cent of the full scale
of the recorder. The test is not valid unless the resolution
between the peaks corresponding to alfuzosin and alfuzosin
impurity A is at least 3.0. Inject 20 l of the test solution and
20 l of reference solution (a). In the chromatogram obtained
with the test solution : the area of any peak, apart from the
principal peak, is not greater than 0.6 times the area of the
principal peak in the chromatogram obtained with reference
solution (a) (0.3 per cent) ; the sum of the areas of all the
peaks, apart from the principal peak, is not greater than the
area of the principal peak in the chromatogram obtained
with reference solution (a) (0.5 per cent). Disregard any peak
with an area less than 0.025 times that of the principal peak
in the chromatogram obtained with reference solution (a).
Injection : 10 l.
Run time : twice the retention time of alfuzosin.
Relative retention with reference to alfuzosin (retention
time = about 8 min) : impurity D = about 0.4 ;
impurity A = about 1.2.
peak-to-valley ratio : minimum 5, 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 alfuzosin.
Limits :
impurity D : not more than twice the area of the principal
(0.3 per cent) ;
(0.05 per cent).
Water (2.5.12). Not more than 2.0 per cent, determined on
0.500 g by the semi-micro determination of water : maximum
0.5 per cent, determined on 1.000 g.
on 1.0 g.
ASSAY
Dissolve 0.300 g in a mixture of 40 ml of anhydrous
acetic acid R and 40 ml of acetic anhydride 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 42.59 mg of
C19H28ClN5O4.
STORAGE
In an airtight container, protected from light.
Figure 1287.-1. Chromatogram for the test for related substances of alfuzosin hydrochloride : test solution spiked
with impurities A, B and D
(2) Inertsil ODS2 is suitable.
242
Avian infectious bursal disease vaccine (inactivated)
IMPURITIES
Reference: PA/PH/Exp. 15V/T (04) 27 ANP
Specified impurities : D.

The monograph has been revised to delete vaccines
grown in chickens from its scope, because no vaccine is
produced in chickens anymore. Therefore, inactivation
test C has also been deleted. This monograph is now
impurities and/or by the general monograph Substances for intended for vaccines used as a booster vaccination or after
pharmaceutical use (2034). It is therefore not necessary to priming with a live vaccine. As the serological threshold
of 10 000 Ph. Eur. U./ml was too strict for current levels
of protection, the potency and immunogenicity tests have
been changed. Furthermore, the monograph is presented
pharmaceutical use) : A, B, C, E.
in the new format to be used in future for all veterinary
vaccines. This editorial re-arrangement does not entail
technical change.
XXXX:0960
AVIAN INFECTIOUS BURSAL DISEASE

VACCINE (INACTIVATED)
Vaccinum bursitidis infectivae aviariae
inactivatum
A. N-[3-[(4-amino-6,7-dimethoxyquinazolin-2yl)(methyl)amino]propyl]furan-2-carboxamide,
B. R-Cl : 2-chloro-6,7-dimethoxyquinazolin-4-amine,
C. (RS)-N-[3-[(4-amino-6,7-dimethoxyquinazolin2-yl)amino]propyl]-N-methyltetrahydrofuran2-carboxamide,
D. N-(4-amino-6,7-dimethoxyquinazolin-2-yl)-Nmethylpropane-1,3-diamine,
E. N-[3-[(4-amino-6,7-dimethoxyquinazolin-2yl)(methyl)amino]propyl]formamide.
1. DEFINITION
Avian infectious bursal disease vaccine (inactivated) consists
of an emulsion or a suspension of a suitable strain of
infectious avian bursal disease virus type 1 which has been
inactivated in such a manner that immunogenic activity
is retained. The vaccine is intended for use in breeding
domestic fowl chickens to protect their progeny from
infectious avian bursal disease.
2. PRODUCTION
2-1. PREPARATION OF THE VACCINE
The virus is propagated grown in fertilised embryonated
hens eggs from healthy flocks, or in suitable cell cultures
(5.2.4) or in chickens from a flock free from specified
pathogens (5.2.2).
The vaccine may contain one or more suitable adjuvants.
2-2. SUBSTRATE FOR VIRUS PROPAGATION
2-2-1. Embryonated hens eggs
If the vaccine virus is grown in embryonated hens eggs,
they are obtained from healthy flocks.
2-2-2. Cell cultures
If the vaccine is grown in cell cultures, they comply with the
requirements for cell cultures for production of veterinary
vaccines (5.2.4).
2-2-3. Chickens
If the vaccine is grown in chickens, they are obtained from a
flock free from specified pathogens (5.2.2).
2-3. CHOICE OF VACCINE COMPOSITION
The vaccine is shown to be satisfactory with respect to safety
(5.2.6) and efficacy (5.2.7). The following test may be used
during demonstration of immunogenicity (2-3-1.).
2-3-1. Immunogenicity. Each of at least twenty A test is
carried out for each route and method of administration to be
recommended using in each case chickens from a flock free
from specified pathogens (SPF) (5.2.2) and not older than the
youngest age to be recommended for vaccination (close to the
point of lay). The quantity of the vaccine virus administered
to each chicken is not greater than the minimum potency
to be stated on the label. Use 2 groups of not less than
20 chickens from a flock free from specified pathogens
243
Avian infectious bursal disease vaccine (inactivated)
(5.2.2), and of the recommended age for vaccination, is

injected with the minimum recommended dose of vaccine by
one of the recommended routes. treated as follows :
group A : unvaccinated controls ;
group B : vaccinated with inactivated avian infectious
bursal disease vaccine.
Serum samples are collected from each chicken just before
administration of the vaccine, 4-6 weeks later and at the time
of egg collection for hatching. serum samples are collected
from each bird and, The antibody response is measured
in a serum-neutralisation (SN) test. A suitable standard,
calibrated in Ph. Eur. Units against infectious avian bursal
disease serum BRP, is included in the test. The vaccine
complies with the test if the mean antibody level in the sera
from the vaccinated birds is at least 10 000 Ph. Eur. Units
per millilitre.
Eggs are collected for hatching 5 to 7 weeks after vaccination
at the end of the period of lay and the test described below is
carried out with 3-week-old at least 15-day-old chickens from
that egg collection. Eggs are collected again towards the end
of the period of lay and the test is repeated with chickens
from that egg collection which are at least 15 days old.
25 chickens from vaccinated (group B) hens and 10 control
chickens of the same breed and age from unvaccinated
(group A) hens are challenged with an eye-drop application
of a quantity of a virulent strain of infectious avian
bursal disease virus sufficient to produce severe signs of
disease, including lesions of the bursa of Fabricius, in all
unvaccinated chickens. 3-4 days after challenge, the bursa
of Fabricius is removed from each chicken. The bursae
are examined for evidence of infection by histological
examination or and by testing for the presence of infectious
avian bursal disease antigen in an agar-gel precipitation test
by a suitable method. The vaccine complies with the test if 3
or fewer of the chickens from vaccinated group B hens show
evidence of infectious avian bursal disease infection. The
test is not valid unless all the chickens from unvaccinated
group A hens are affected show evidence of infectious avian
bursal disease infection.
Where there is more than one recommended route of
administration, the test described under Potency is carried
out in parallel with the above immunogenicity test, using
different groups of birds for each recommended route. The
serological response of the birds inoculated by routes other
than that used in the immunogenicity test is not significantly
less than that of the group vaccinated by that route.
2-4. MANUFACTURERS TESTS
2-4-1. Inactivation
An amplification test for residual live infectious avian
bursal disease virus is carried out on each batch of antigen
immediately after inactivation and on the final bulk vaccine
or, if the vaccine contains an adjuvant, on the bulk antigen
or the mixture of bulk antigens immediately before the
addition of any adjuvant, to confirm inactivation ; the
test is carried out in fertilised hen eggs or in suitable cell
cultures or, where chickens have been used for production
of the vaccine, in chickens from a flock free from specified
pathogens (5.2.2) ; the quantity of inactivated virus used in
the test is equivalent to not less than 10 doses of the vaccine.
No live virus is detected.
2-4-2. Batch potency test
Where the test described below is not carried out, an
alternative validated method is used, the criteria for
acceptance being set with reference to a batch of vaccine
that has given satisfactory results in the test described under
244
Immunogenicity (section 2-3-1.). The following test may be

used. Vaccinate each of not less than 10 chickens, 4 weeks of
age and from an SPF flock (5.2.2), with one dose of vaccine
by one of the recommended routes. 4-6 weeks later, collect
serum samples from each bird and 10 unvaccinated control
birds of the same age and from the same source. Measure the
antibody response in an SN test. Include in the test a suitable
standard, calibrated in Ph.Eur. Units against infectious
avian bursal disease serum BRP. The vaccine complies with
the test if the mean antibody level titre in the sera from
the vaccinated birds is not less than 10 000 Ph. Eur. Units
per millilitre equal to or greater than the titres obtained
with a batch that has given satisfactory results in the test
described under Immunogenicity (section 2-3-1.). There are
no antibodies in the sera of the unvaccinated birds.
3. BATCH TESTS
3-1. Identification
In When injected into chickens with no free from antibodies
to against infectious avian bursal disease virus type 1, the
vaccine stimulates the production of specific such antibodies.
3-2. Bacteria and fungi
The vaccine complies with the test for sterility prescribed in
the monograph on Vaccines for veterinary use (0062).
3-3. Extraneous agents
Use 10 chickens, 14-28 days old from an SPF flock (5.2.2).
Vaccinate each chicken by a recommended route with a
double dose of the vaccine. After 3 weeks, inject 1 dose by
the same route. Collect serum samples from each of the
birds used in the safety test or inactivation test C chicken
three 2 weeks after vaccination later and carry out tests
for antibodies to the following agents by the methods
prescribed in chapter 5.2.2 : avian encephalomyelitis virus,
avian leucosis viruses, haemagglutinating avian adenovirus,
egg-drop syndrome virus, avian infectious bronchitis virus,
avian infectious laryngotracheitis virus, influenza A virus,
Mareks disease virus, Newcastle disease virus. The vaccine
does not stimulate the formation of antibodies against these
agents.
3-4. Safety
Use not less than 10 chickens, 14-28 days old, from SPF
flocks (5.2.2). Administer to each chicken by a recommended
route a double dose of vaccine. Observe the birds at least
daily for not less than 21 days. No abnormal local or
systemic reaction occurs. Note : this test may be omitted if
inactivation test C is carried out. The vaccine complies with
the test if no bird shows notable signs of disease or dies from
causes attributable to the vaccine.
3-5. Inactivation
A. For vaccine prepared with embryo-adapted strains of
virus, inject 2/5 of a dose into the allantoic cavity or onto
the chorio-allantoic membrane of ten 9- to 11-day-old
fertilised hen eggs from an SPF flock (SPF eggs)
(5.2.2). Incubate the eggs and observe for 6 days. Pool
separately the allantoic liquid or membranes from eggs
containing live embryos, and that from eggs containing
dead embryos, excluding those that die from non-specific
causes within 24 h of the injection.
Inject into the allantoic cavity or onto the chorio-allantoic
membrane of each of ten 9- to 11-day-old SPF eggs 0.2 ml
of the pooled allantoic liquid or crushed chorio-allantoic
membranes from the live embryos and, into each of
10 similar eggs, 0.2 ml of the pooled liquid or membranes
from the dead embryos and incubate for 6 days. Examine
each embryo for lesions of infectious avian bursal disease.
Cholera vaccine (inactivated, oral)
The production process must be validated to show that no

clinically signifiant quantities of active toxin are present in
the product.
CHOICE OF VACCINE STRAIN
The vaccine consists of a mixture of epidemic V. cholerae
strains inactivated by a suitable method such as heat
or formalin inactivation. All strains express smooth
lipopolysaccharide (LPS). The CTB is produced by
recombinant DNA technology in a strain that lacks the gene
for cholera toxin subunit A (ctxA-). Selected V. cholerae
strains are low cholera-toxin producers.
The World Health Organisation (WHO) can recommend new
vaccine strains or antigens that may be used if necessary, in
accordance with the regulations in force in the signatory
states of the Convention on the Elaboration of a European
Pharmacopoeia.
SEED LOTS
The strains of V. cholerae used shall be identified by historical
records that include information on the origin of the strains
and their subsequent manipulation. Characterisation and
maintenance of the recombinant strain and plasmids used
for production of the recombinant B subunit of cholera
toxin (rCTB) and the origin of the gene for cholera toxin
subunit B (ctxB) are documented. The stability of the rCTB
plasmid in the recombinant strain during storage and beyond
the passage level used in production is confirmed.
Characterisation of the rCTB is undertaken using a variety of
analytical techniques including determination of molecular
size, charge and amino acid composition. Techniques
suitable for such purposes include sodium dodecyl sulphate
polyacrylamide gel electrophoresis (SDS-PAGE) and different
liquid chromatographies. The identity of the product is
confirmed by at least partial N-terminal and C-terminal
amino acid sequencing.
Master seed lots are grown on agar plates, which may
contain appropriate antibiotics. Colonies are used to
produce working seed lots in liquid media that are free from
4. LABELLING
antibiotics. Cultures derived from the working seed lot must
The label states whether the strain in the vaccine is
have the same characteristics as the cultures of the strain
embryo-adapted or cell-culture-adapted.
from which the master seed lot was derived.
Only a seed lot that complies with the following requirements
may be used in the preparation of the monovalent cell
harvest.
Reference: PA/PH/Exp. 15/T (05) 23 ANP
Identification. Master seed lots are identified by colony
XXXX:2327 morphology, biochemical characterisation, molecular assays
or immunoassays. Working seed lots are identified by colony
morphology and by molecular assays or immunoassays.
CHOLERA VACCINE (INACTIVATED, Purity. Purity of master seed lots and working seed lots is
ORAL)
verified by methods of suitable sensitivity.
PROPAGATION AND HARVEST
Vaccinum cholerae perorale inactivatum Each strain is grown separately from the working seed lot.
Cultures are checked at different stages of fermentation
DEFINITION
(subcultures and main culture) for purity, identity, cell
Cholera vaccine (inactivated, oral) is a homogeneous
suspension of inactivated suitable strains of Vibrio cholerae opacity, pH and biochemical characteristics. Unsatisfactory
cultures must be discarded.
serogroup O1 representing serotypes and biotypes of
Production cultures are shown to be consistent in respect of
epidemic strains. The vaccine may contain the B subunit
growth rate, pH and yield of cells or cell products.
of cholera toxin (CTB). Just prior to ingestion, one dose of
vaccine suspension is mixed with a suitable buffer as stated MONOVALENT CELL HARVEST
on the label.
Only a monovalent harvest that complies with established
specifications for the following tests may be used.
PRODUCTION
pH (2.2.3) : within the range approved for the particular
GENERAL PROVISIONS
product.
The production method must be validated to yield
consistently vaccines comparable with the vaccine of proven Identification. Relevant antigenic characteristics are verified
clinical efficacy and safety in man.
by suitable immunological or biochemical assays.
If more than 20 per cent of the embryos die at either
stage repeat that stage. The vaccine complies with the
test if there is no evidence of lesions of infectious avian
bursal disease and if, in any repeat test, not more than
20 per cent of the embryos die from non-specific causes.
Antibiotics may be used in the test to control extraneous
bacterial infection.
B. For vaccine prepared with cell-culture-adapted strains
of virus, inoculate 10 doses of the vaccine into suitable
cell cultures. If the vaccine contains an oil adjuvant,
eliminate it by suitable means. Incubate at 38 1 C for
7 days. Make a passage on another set of cell cultures
and incubate at 38 1 C for 7 days. The cultures show
no signs of infection.
C. For vaccine prepared with strains of virus not adapted to
embryos or cell cultures, inject two doses intramuscularly
into each of twenty 14- to 28-day-old chickens from flocks
free from specified pathogens (5.2.2). 4 days later, kill
ten of the chickens and remove the bursa of Fabricius
from each chicken. Pool the bursae and homogenise in
an equal volume of a suitable liquid. Inject 1 ml of the
bursal homogenate into each of a further ten chickens
of the same age and from the same source. Examine
microscopically the bursa of Fabricius of each remaining
chicken from the first group and of each chicken from
the second group 21 days after the injection ; there is no
evidence of infectious avian bursal disease infection. In
addition there are no signs of any disease attributable
to the vaccine and no abnormal local reaction develops.
The chickens of the second group do not have antibodies
against infectious avian bursal disease virus when
examined 21 days after the injection.
3-6. Potency
The vaccine complies with the requirements of the test
mentioned under Immunogenicity (section 2-3-1.) using
1 dose of the vaccine administered by a recommended route.
245
Cholera vaccine (inactivated, oral)
Purity. Samples of culture are examined by microscopy of

Gram-stained smears, by inoculation of appropriate culture
media or by another suitable procedure.
Opacity. The absorbance at 600 nm (2.2.25) is within the
range approved for the particular product.
INACTIVATED MONOVALENT CELL BULK
To limit the possibility of contamination, inactivation is
initiated as soon as possible after preparation. Bacteria
are inactivated after washing, either by treatment with
formaldehyde or by heating under conditions that ensure
inactivation.
Only an inactivated monovalent cell bulk that complies with
established specifications for the following tests may be used
in the preparation of the final bulk.
product.
Identification : verified by slide agglutination.
Inactivation. Complete inactivation is verified by a suitable
culture method.
Sterility (2.6.1). It complies with the test for sterility, carried
out using 10 ml for each medium.
Opacity. The inactivation process may affect the accuracy of
opacity measurements.
Purity. Samples of culture are examined by microscopy of
Gram-stained smears, by inoculation of appropriate culture
media or other suitable procedure.
Smooth LPS content : verified by a suitable immunoassay
(2.7.1).
Residual cholera toxin. The absence of cholera toxin is
verified by a suitable immunoassay (2.7.1) or biochemical
assay.
Free formaldehyde (2.4.18) : to be determined where
formaldehyde is used for inactivation.
PURIFIED rCTB
Production of the rCTB follows the guidelines for assuring
the quality of pharmaceutical and biological products
prepared by recombinant technology. Prior to harvest,
the cell culture is checked for purity and opacity. rCTB is
harvested by suitable filtration, concentrated by diafiltration,
purified by chromatography, filter-sterilised and stored
under suitable conditions. The pH of the pooled eluate is
adjusted prior to buffer exchange.
Only purified rCTB that complies with established
specifications for the following tests may be used in the
preparation of the final bulk.
product.
Purity : verified by SDS-PAGE (2.2.31) and an appropriate
liquid chromatography method (2.2.29).
246
rCTB. The amount of rCTB is determined by a suitable

immunoassay (2.7.1).
FINAL BULK
The final bulk vaccine is prepared by aseptically mixing
suitable buffer with monovalent cell bulks. Where used, the
rCTB bulk is added in appropriate amounts. Preservative, if
used, may be added at this stage.
Only a final bulk that complies with the following
requirements may be used in the preparation of the final lot.
Antimicrobial preservative. Where applicable, determine
the amount of antimicrobial preservative by a suitable
chemical or physico-chemical method. The amount is not
less than 85 per cent and not greater then 115 per cent of
the intended amount.
FINAL LOT
The final bulk is mixed to homogeneity and filled asceptically
into suitable containers.
Only a final lot that is within the limits approved for the
particular product and is satisfactory with respect to each of
the requirements given below under Identification, Tests and
Assay may be released for use.
IDENTIFICATION
Serotypes are detected by a suitable immunoassay (2.7.1) or
molecular assay. rCTB is detected by a suitable immunoassay
(2.7.1). The antigen content assays may also serve as an
identity test.
TESTS
product.
Sterility (2.6.1). It complies with the test for sterility.
Free formaldehyde (2.4.18) : maximum 0.2 g/l, where
applicable.
Antimicrobial preservative. Where applicable, determine
the amount of antimicrobial preservative by a suitable
chemical or physico-chemical method. The amount is not
less than 85 per cent and not greater than 115 per cent of
the intended amount.
ASSAY
Antigen content. The amount of smooth LPS, and rCTB
where applicable, is determined by a suitable immunoassay
(2.7.1).
LABELLING
The label states :
the method used for inactivation ;
the serogroup, serotypes and biotypes of vaccine strains ;
the number of bacteria per human dose ;
the amount of rCTB.
Clopamide
Reference: PA/PH/Exp. 10C/T (98) 11 ANP 3R

The monograph proposal published in Pharmeuropa 11.1
has been completely reviewed. It now inludes a new LC
method that covers additional impurities. A GC method has
been introduced to control impurity D. Its limit has been
set up using the TTC (Threshold of Toxicological Concern)
approach : a tolerated daily dose of 1.5 g/day corresponds
to a limit of 0.03 per cent for a 5 mg daily intake of
clopamide. The immediate precursor of impurity D is an
N-nitroso compound (cis-2,6-dimethyl-1-nitroso-piperidine).
Limiting impurity D to a low level allows the indirect
limitation of the potentially toxic N-nitroso compound to
an even lower level.
XXXX:1747
Solubility : slightly soluble in water, sparingly soluble in

methanol, slightly soluble in anhydrous ethanol.
It shows polymorphism.
IDENTIFICATION
Comparison : clopamide CRS.
If the spectra obtained in the solid state show differences,
dissolve the substance to be examined and the reference
substance separately in the minimum volume of methanol R,
evaporate to dryness on a water-bath and record new spectra
using the residues.
TESTS
Test solution. Dissolve 100.0 mg of the substance to be
examined in methanol R and dilute to 10.0 ml with the same
solvent.
CLOPAMIDE
Reference solution (a). Dissolve 10.0 mg of clopamide for
system suitability CRS (containing impurities A, B, C and H)
Clopamidum
in methanol R and dilute to 1.0 ml with the same solvent.
Reference solution (b). Dilute 2.0 ml of the test solution to
100.0 ml with methanol R. Dilute 2.0 ml of this solution to
20.0 ml with methanol R.
Reference solution (c). Dilute 2.5 ml of reference solution (b)
to 10.0 ml with methanol R.
Column :
size : l = 0.15 m, = 4.6 mm ;
C14H20ClN3O3S
Mr 345.8 stationary phase : octylsilyl silica gel for
chromatography R (5 m)(3) ;
DEFINITION
temperature : 28 C.
4-Chloro-N-[(2RS,6SR)-2,6-dimethylpiperidin-1-yl]-3Mobile
phase :
sulfamoylbenzamide.
mobile
phase A : dissolve 1.0 g of ammonium acetate R
in 950 ml of water R, adjust to pH 2.0 with concentrated
phosphoric acid R and dilute to 1000 ml with water R ;
CHARACTERS
mobile
phase B : acetonitrile R ;
Appearance : white or almost white, hygroscopic, crystalline
mobile phase C : water R, tetrahydrofuran R (20:80 V/V) ;
powder.
1. impurity C
2. impurity G
3. clopamide
4. impurity H
5. impurity B
Figure 1747.-1. Chromatogram for the test for related substances of clopamide
(3) Zorbax Eclipse XDB-C8 5 m is suitable.
247
Clopamide
Time
(min)
0 - 35
Mobile phase A
(per cent V/V)
95 75
Mobile phase B
(per cent V/V)
5 25
Mobile phase C
(per cent V/V)
0
35 - 45
75 35
25 65
45 - 50
35 30
65 0
0 70
50 - 60
30
70
60 - 63
30 95
05
70 0
63 - 73
95

Injection: 10 l.
Identification of impurities : use the chromatogram
supplied with clopamide for system suitability CRS and
the chromatogram obtained with reference solution (a) to
identify the peaks due to impurities A, B, C and H.
Relative retention with reference to clopamide (retention
time = about 33.3 min) : impurity C = about 0.81 ;
impurity G = about 0.84 ; impurity A = about 0.92 ;
impurity H = about 1.23 ; impurity B = about 1.43.
System suitability : reference solution (a) :
impurity A and clopamide.
Limits :
the corresponding correction factor : impurity B = 0.4 ;
impurity C = 0.7 ; impurity H = 0.4 ;
impurities A, B, C, H : for each impurity, not more than
the area of the principal peak in the chromatogram
obtained with reference solution (b) (0.2 per cent) ;
impurity G : not more than 0.5 times the area of the

principal peak in the chromatogram obtained with
reference solution (b) (0.1 per cent) ;
than 0.5 times the area of the principal peak in the
chromatogram obtained with reference solution (b)
(0.10 per cent) ;
in the chromatogram obtained with reference solution (b)
(1.0 per cent) ;
disregard limit : the area of the principal peak in the
chromatogram obtained with reference solution (c)
(0.05 per cent).
Impurity D. Gas chromatography (2.2.28).
examined in methanol R and dilute to 10.0 ml with the same
solvent.
Reference solution (a). Dissolve 5.0 mg of clopamide
impurity D CRS in methanol R and dilute to 10.0 ml with
the same solvent. Dilute 5.0 ml of this solution to 100.0 ml
with methanol R.
Reference solution (b). Dilute 1.0 ml of reference solution (a)
to 10.0 ml with methanol R.
Column :
material : fused silica ;
size : l = 30 m, = 0.32 mm ;
stationary phase : poly(dimethyl)(diphenyl)siloxane R1(4)
(film thickness 0.25 m).
Carrier gas : helium for chromatography R.
Split ratio : 1:10.
1. impurity C
2. impurity A
3. clopamide
4. impurity B
Figure 1747.-2. Chromatogram for the test for related substances of clopamide
(4) SPB 20 (Supelco) is suitable.
248
Clozapine
Temperature :
Column
Injection port
Time
(min)
0-1
Temperature
(C)
60
1 - 10
60 150
10 - 13.75
150 300
13.75 - 23.75
300
0 - 12
200
12 - 13
200 300
13 - 18
300
Detector
D. (2RS,6SR)-2,6-dimethylpiperidin-1-amine,
300
Detection : flame ionisation.

Injection: 1 l.
Retention time : impurity D = about 4.8 min.
signal-to-noise ratio : minimum 10 for the principal peak.
Limit :
impurity D : not more than 0.3 times the area of the
corresponding peak in the chromatogram obtained with
reference solution (a) (0.03 per cent).
Heavy metals (2.4.8) : maximum 20 ppm.
1.0 g complies with test C. Prepare the reference solution
using 2 ml of lead standard solution (10 ppm Pb) R.
on 1.000 g by drying in an oven at 105 C.
on 1.0 g.
G. 3-(aminosulfonyl)-4-chloro-N-(2-methylpiperidin-1yl)benzamide,
H. 4-chloro-3-[[[(1Z)-(dimethylamino)methylene]amino]sulfonyl]-N-(2,6-dimethylpiperidin-1-yl)benzamide.
Reagents
Poly(dimethyl)(diphenyl)siloxane R1. XXXXXXX.

ASSAY
Stationary phase for gas chromatography.
Dissolve 0.280 g in 70 ml of anhydrous acetic acid R. Titrate Contains 80 per cent of methyl groups and 20 per cent of
phenyl groups.
of C14H20ClN3O3S.
STORAGE
It is proposed to replace the TLC test for related substances
IMPURITIES
with an LC test. Only comments on this test are requested.
XXXX:1191
Specified impurities : A, B, C, D, G, H.
CLOZAPINE
Clozapinum
A. 4-chloro-N-[(2RS,6RS)-2,6-dimethylpiperidin-1-yl]-3sulfamoylbenzamide (trans-clopamide),
C18H19ClN4
B. R = H : 4-chlorobenzoic acid,
C. R = SO2-NH2 : 4-chloro-3-sulfamoylbenzoic acid,
Mr 326.8
DEFINITION
8-Chloro-11-(4-methylpiperazin-1-yl)-5H-dibenzo[b,e][1,
4]diazepine.
249
Clozapine
CHARACTERS
Appearance : yellow, crystalline powder.
Solubility : practically insoluble in water, freely soluble in
methylene chloride, soluble in ethanol (96 per cent). It
dissolves in dilute acetic acid.
IDENTIFICATION
A. Melting point (2.2.14) : 182 C to 186 C.
B. Infrared absorption spectrophotometry (2.2.24).
Comparison : clozapine CRS.
Examine the substances prepared as discs.
TESTS
(2.2.27), using as the coating substance a suitable silica gel
with a fluorescent indicator having an optimal intensity at
254 nm. Before use, develop the plate with a mixture of
25 volumes of methanol R and 75 volumes of methylene
chloride R. Allow the plate to dry in air for 15 min.
Test solution (a). Dissolve 0.20 g of the substance to be
examined in methylene chloride R and dilute to 5 ml with
the same solvent.
Test solution (b). Dilute 1 ml of test solution (a) to 10 ml
with methylene chloride R.
Reference solution (a). Dissolve 20 mg of clozapine CRS
in methylene chloride R and dilute to 5 ml with the same
solvent.
Reference solution (b). Dilute 1.5 ml of test solution (b) to
50 ml with methylene chloride R.
Reference solution (c). Dilute 1 ml of test solution (b) to
50 ml with methylene chloride R.
Reference solution (d). Dilute 5 ml of reference solution (c)
to 10 ml with methylene chloride R.
Reference solution (e). Dissolve 10 mg of clozapine CRS
and 10 mg of oxazepam CRS in methylene chloride R and
dilute to 5 ml with the same solvent.
Apply separately to the plate 5 l of each solution. Develop

over a path of 10 cm using a mixture of 25 volumes of
methanol R and 75 volumes of methylene chloride R. Allow
the plate to dry in air and examine in ultraviolet light at
254 nm. Any spot in the chromatogram obtained with test
solution (a), apart from the principal spot, is not more intense
than the spot in the chromatogram obtained with reference
solution (b) (0.3 per cent) ; at most one such spot is more
intense than the spot in the chromatogram obtained with
reference solution (c) (0.2 per cent) ; and at most two such
spots are more intense than the spot in the chromatogram
obtained with reference solution (d) (0.1 per cent). The
test is not valid unless the chromatogram obtained with
reference solution (e) shows two clearly separated spots
and the chromatogram obtained with reference solution (d)
shows a clearly visible spot.
Solvent mixture : water R, methanol R2 (20:80 V/V).
Solution A. Dissolve about 2.04 g of potassium dihydrogen
phosphate R in 1000 ml of water R and adjust to pH 2.4
with phosphoric acid R. The pH of this solution must not
be below 2.4.
examined in 80 ml of methanol R2 and dilute to 100.0 ml
with water R.
to 10.0 ml with the solvent mixture. Dilute 1.0 ml of this
solution to 100.0 ml with the solvent mixture.
Reference solution (b). Dissolve 4 mg of clozapine for peak
identification CRS (containing impurities A, B, C and D) in
4 ml of methanol R2, add 1 ml of the test solution and 1 ml
of water R and dilute to 10 ml with the solvent mixture.
Column :
size : l = 0.125 m, = 4.6 mm ;
for chromatography R (5 m)(5).
1. impurity C
2. clozapine
3. impurity D
4. impurity A
5. impurity B
Figure 1191.-1. Chromatogram for the test for related substances of clozapine : solution of clozapine spiked with
impurities A, B, C and D
(5) Nucleosil 100-5 C18 HD, YMC-Pack ODS-AQ S-5 (0.15 m 4.6 mm ; 5 m), Inertsil ODS3 and Kromasil RP (0.125 m 4.0 mm ; 5 m) are suitable.
250
Clozapine
Mobile phase :
ASSAY
mobile phase A : acetonitrile for chromatography R,

methanol R2, solution A (1:1:8 V/V/V) ;
Dissolve 0.100 g in 50 ml of anhydrous acetic acid R. Titrate

mobile phase B : solution A, acetonitrile for

chromatography R, methanol R2 (2:4:4 V/V/V) ;

of C18H19ClN4.
Time
(min)
0-4
Mobile phase A
(per cent V/V)
100
Mobile phase B
(per cent V/V)
0
4 - 24
100 0
0 100
24 - 29
100
IMPURITIES
Specified impurities : A, B, C, D.

Injection : 20 l.
Run time : 2.5 times the retention time of clozapine.
Relative retention with reference to clozapine (retention
time = about 11 min) : impurity C = about 0.9 ;
impurity D = about 1.1 ; impurity A = about 1.6 ;
impurity B = about 1.7.
A. 8-chloro-5,10-dihydro-11H-dibenzo[b,e][1,4]diazepin-11one,

impurity C and clozapine.
Limits :
correction factor : for the calculation of content, multiply
the peak area of impurity D by 2.7 ;
impurity A : not more than the area of the principal peak
(0.1 per cent) ;
B. 11,11-(piperazine-1,4-diyl)bis(8-chloro-5H-dibenzo[b,e][1,
4]diazepine),
impurities B, D : for each impurity, not more than twice
impurity C : not more than 3 times the area of the
reference solution (a) (0.3 per cent) ;
total: not more than 6 times the area of the principal peak C. 8-chloro-11-(piperazin-1-yl)-5H-dibenzo[b,e][1,4]diazepine,
(0.6 per cent) ;
(0.05 per cent) ; disregard any peak due to the blank.
on 1.0 g.
D. 4-chloro-N1-[2-[(4-methylpiperazin-1-yl)carbonyl]phenyl]benzene-1,2-diamine.
251
Copovidone
pH (2.2.3) : 3.0 to 7.0 for solution S.

Appearance of solution. Solution S is not more opalescent
than reference suspension III (2.2.1) and not more intensely
The revision proposal for this monograph is published
coloured than reference solution B5, R5 or BY5 (2.2.2,
below as part of the framework of harmonisation with the Method II).
JP and the USP. The stage 4 draft provided by the JP is
Aldehydes : maximum 500 ppm, expressed as acetaldehyde.
published in the International harmonisation section.
XXXX:0891 Test solution. Dissolve 1.0 g of the substance to be examined
in phosphate buffer solution pH 9.0 R and dilute to 100.0 ml
with the same solvent. Tightly stopper the flask and heat at
COPOVIDONE
60 C for 1 h. Allow to cool.
Reference solution. Dissolve 0.140 g of acetaldehyde
Copovidonum
ammonia trimer trihydrate R in water R and dilute to
200.0 ml with the same solvent. Dilute 1.0 ml of this solution
to 100.0 ml with phosphate buffer solution pH 9.0 R.
Into 3 identical spectrophotometric cells with a path length
of 1 cm, introduce separately 0.5 ml of the test solution,
0.5 ml of the reference solution and 0.5 ml of water R
(blank). To each cell, add 2.5 ml of phosphate buffer
solution pH 9.0 R and 0.2 ml of nicotinamide-adenine
dinucleotide solution R. Mix and stopper tightly. Allow to
stand at 22 2 C for 2-3 min and measure the absorbance
(C6H9NO)n, (C4H6O2)m
Mr (111.1)n + (86.1)m (2.2.25) of each solution at 340 nm, using water R as the
compensation liquid. To each cell, add 0.05 ml of aldehyde
dehydrogenase solution R, mix and stopper tightly. Allow
DEFINITION
to stand at 22 2 C for 5 min. Measure the absorbance of
Copolymer of 1-ethenylpyrrolidin-2-one and ethenyl acetate each solution at 340 nm using water R as the compensation
in the mass proportion 3:2.
liquid. Determine the content of aldehydes using the
Content :
following expression :
nitrogen (N ; Ar 14.01) : 7.0 per cent to 8.0 per cent (dried
substance) ;
ethenyl acetate (C4H6O2 ; Mr 86.10) : 35.3 per cent to
42.0 per cent (dried substance).
K-value : 90.0 per cent to 110.0 per cent of the value stated At1 = absorbance of the test solution before the addition
on the label.
of aldehyde dehydrogenase ;
At2 = absorbance of the test solution after the addition
CHARACTERS
of aldehyde dehydrogenase ;
Appearance : white or yellowish-white, hygroscopic powder
As1 = absorbance of the reference solution before the
or flakes.
addition of aldehyde dehydrogenase ;
Solubility : freely soluble in water, in ethanol (96 per cent)
As2 = absorbance of the reference solution after the
and in methylene chloride.
addition of aldehyde dehydrogenase ;
IDENTIFICATION
Ab1 = absorbance of the blank before the addition of
aldehyde dehydrogenase ;
First identification : A.
A
Second identification : B, C.
= absorbance of the blank after the addition of
b2
aldehyde dehydrogenase ;
m
= mass of povidone copovidone, in grams, calculated
Preparation : discs.
with reference to the dried substance ;
Dry the substance to be examined and the reference
C
concentration (mg/ml) of acetaldehyde in the
=
substance at 105 C for 3 h.
reference solution, calculated from the weight of
Comparison : Ph. Eur. reference spectrum of
the acetaldehyde ammonia trimer trihydrate with
copovidone.
the factor 0.72.
B. To 1 ml of solution S (see Tests) add 5 ml of water R and
Peroxides : maximum 400 ppm, expressed as H2O2.
0.2 ml of 0.05 M iodine. A red colour appears.
10 ml of solution S to 25 ml with water R. Prepare a
Dilute
C. Dissolve 0.7 g of hydroxylamine hydrochloride R in
solution
containing 1.0 g of dried copovidone in 25 ml of
10 ml of methanol R, add 20 ml of a 40 g/l solution of
water
R.
Add 2 ml of titanium trichloride-sulphuric acid
sodium hydroxide R and filter if necessary. To 5 ml of
reagent R and allow to stand for 30 min. The absorbance
this solution add 0.1 g of the substance to be examined
and boil for 2 min. Transfer 50 l to a filter paper and add (2.2.25) of the solution, measured at 405 nm using a mixture
of 25 ml of a 40 g/l solution of the substance to be examined
0.1 ml of a mixture of equal volumes of ferric chloride
and 2 ml of a 13 per cent V/V solution of sulphuric acid R
solution R1 and hydrochloric acid R. A violet colour
as the compensation liquid, is not greater than 0.35.
appears.
Hydrazine. Thin-layer chromatography (2.2.27). Use freshly
TESTS
prepared solutions.
Solution S. Dissolve 10 g in water R, adding the substance
Test solution. To 25 ml of solution S add 0.5 ml of a 50 g/l
to be examined in small portions with constant stirring and solution of salicylaldehyde R in methanol R, mix and heat
dilute to 100 ml with the same solvent.
in a water-bath at 60 C for 15 min. Allow to cool, add 2.0 ml
252
Copovidone
of xylene R toluene R, shake for 2 min and centrifuge. Use

the clear supernatant layer.
Reference solution. Dissolve 9 mg of salicylaldehyde
azine R in xylene R toluene R and dilute to 100 ml with
the same solvent. Dilute 1 ml of this solution to 10 ml with
xylene R toluene R.
Plate : TLC silanised silica gel plate R.
Mobile phase : water R, methanol R (20:80 1:2 V/V).
Application : 10 l.
Development : over a path of 15 cm.
Drying : in air.
Detection : examine in ultraviolet light at 365 nm.
Results : the Rf value of the fluorescent spot in the
chromatogram obtained with the reference solution is
about 0.3
Limit :
hydrazine : any spot corresponding to salicylaldehyde
azine in the chromatogram obtained with the test solution
is not more intense than the spot in the chromatogram
obtained with the reference solution (1 ppm).
Monomers : maximum 0.1 per cent.
Dissolve 10.0 g in 30 ml of methanol R and add slowly
20.0 ml of iodine bromide solution R. Allow to stand for
30 min protected from light with repeated shaking. Add
10 ml of a 100 g/l solution of potassium iodide R and titrate
with 0.1 M sodium thiosulphate until a yellow colour is
obtained. Continue titration dropwise until the solution
becomes colourless. Carry out a blank titration. Not more
than 1.8 ml of 0.1 M sodium thiosulphate is used.
Impurity A. Liquid chromatography (2.2.29).
examined in water R and dilute to 50.0 ml with the same
solvent.
Reference solution. Dissolve 100 mg of 2-pyrrolidone R in
water R and dilute to 100 ml with the same solvent. Dilute
1.0 ml to 100.0 ml with water R.
Precolumn :
size : l = 0.025 m, = 4 mm,
for chromatography R (5 m).(6)
Column :
size : l = 0.25 m, = 4 mm,
stationary phase : spherical aminohexadecylsilyl silica
gel for chromatography R (5 m)(7),
temperature : 30 C.
Mobile phase : water R, adjusted to pH 2.4 with phosphoric
acid R.
Flow rate : 1 ml/min.
Detection : spectrophotometer at 205 nm. A detector is
placed between the precolumn and the analytical column. A
second detector is placed after the analytical column.
Injection : 10 l. When impurity A has left the precolumn
(after about 1.2 min) switch the flow directly from the pump
to the analytical column. Before the next chromatogram is
run, wash the precolumn by reversed flow.
Limit :
impurity A : not more than the area of the principal peak
obtained with the reference solution (0.5 per cent).
(6)
(7)
(8)
(9)
Monomers (impurities B and C) and impurity A. Liquid

chromatography (2.2.29).
examined in 1 ml of methanol R, sonicate and dilute
to 10.0 ml with water R. Filter if necessary to remove
undissolved particles.
Reference solution. Dissolve 50 mg of 1-vinyl-2pyrrolidone R (impurity B), 50 mg of vinyl acetate R
(impurity C) and 300 mg of 2-pyrrolidone R (impurity A) in
methanol R and dilute to 100.0 ml with the same solvent.
Dilute 1.0 ml of the solution to 100.0 ml with mobile phase A.
Dilute 5 ml of this solution to 100.0 ml with mobile phase A.
Precolumn :
size : l = 0.025 m, = 4 mm ;
Column :
size : l = 0.25 m, = 4 mm ;
temperature : 30 C.
Mobile phase :
mobile phase A : methanol R, acetonitrile R, water R
(5:5:90) ;
mobile phase B : methanol R, acetonitrile R, water R
(5:45:50).
Time
(min)
0-2
Mobile phase A
(per cent V/V)
100
Mobile phase B
(per cent V/V)
0
2 - 26
100 80
0 20
26 - 27
80 0
20 100
27 - 36
100
36 - 38
0 100
100 0

Detection : spectrophotometer at 205 nm and 235 nm.
Injection : 10 l. After each injection of the test solution,
wash the polymeric material of copovidone from the guard
column by passing the mobile phase through the column
backwards for about 30 min at the same flow rate as applied
in the test.
Elution order : impurity A, impurity C, impurity B.
System suitability : reference solution :
impurity A and impurity C, and to impurity C and
impurity B ;
repeatability : maximum relative standard deviation of
2.0 per cent for each peak after 6 injections.
Limits :
impurity A : not more than the area of the corresponding
peak in the chromatogram obtained with the reference
solution (0.5 per cent) ;
impurities B, C : for each impurity, not more than the area
of the corresponding peak in the chromatogram obtained
with the reference solution (10 ppm),
12 ml of solution S complies with limit test A. Prepare the
standard using lead standard solution (2 ppm Pb) R.
Lichrospher 100 RP 18E is suitable.

Supelcosil ABZ plus is suitable.
Nucleosil 120-5 C18 is suitable.
Aquasil C18 (thermo hypersil) is suitable.
253
2.2.17. Drop point

on 0.500 g by drying in an oven at 100-105 C for 3 h.
on 1 g.
Viscosity, expressed as K-value : 90.0 per cent to 110.0 per
cent of the nominal K-value.
Dilute 5.0 ml of solution S to 50.0 ml with water R. Allow
to stand for 1 h and determine the viscosity (2.2.9) of the
solution at 25 0.1 C using viscometer No. 1 with a
minimum flow time of 100 s. Calculate the K-value from the
IMPURITIES
Specified impurities : A, B, C.
A. pyrrolidin-2-one (2-pyrrolidone),
B. 1-vinylpyrrolidin-2-one,
c
=
=
percentage concentration (g/100 ml) of the

substance to be examined, calculated with
reference to the dried substance ;
viscosity of the solution relative to that of water.
ASSAY
Ethenyl acetate. Determine the saponification value (2.5.6)
on 2.00 g of the substance to be examined. Multiply the
result obtained by 0.1534 to obtain the percentage content
of the ethenyl acetate component.
Nitrogen. Carry out the determination of nitrogen (2.5.9)
using 30.0 mg of the substance to be examined and 1 g of
a mixture of 3 parts of copper sulphate R and 997 parts of
dipotassium sulphate R, heating until a clear, light green
solution is obtained and then for a further 45 min.
Nitrogen. Weigh 0.1 g in a combustion flask. Add 5 g of
a powdered mixture of 33 g of dipotassium sulfate R, 1 g
of copper sulfate R and 1 g of titanium dioxide R, and
wash down any adhering particles from the neck of the
flask with a small amount of water R. Add 7 ml of sulfuric
acid R, allowing it to flow down the inside wall of the flask.
Heat the flask gradually until the solution has a clear,
yellow-green colour, and the inside wall of the flask is free
from any carbonised material. Heat for a further 45 min.
After cooling, cautiously add 20 ml of water R, and connect
the flask to the distillation apparatus previously washed
by passing steam through it. To the absorption flask add
30 ml of a 4 per cent V/V solution of boric acid R, 3 drops
of bromocresol green-methyl red solution R and sufficient
water R to immerse the lower end of the condenser tube.
Add 30 ml of strong sodium hydroxide solution R through a
funnel. Cautiously rinse the funnel with 10 ml of water R,
immediately close the clamp attached to the rubber tube,
then start the distillation with steam to obtain 80-100 ml
of distillate. Remove the absorption flask from the lower
end of the condenser tube, rinsing the end part with a
small quantity of water R, and titrate the distillate with
0.025 M sulfuric acid until the colour of the solution
changes from green through pale greyish blue to pale greyish
red-purple. Carry out a blank determination, and make any
necessary correction.
1 ml of 0.025 M sulfuric acid is equivalent to 0.7004 mg of N.
STORAGE
In an airtight container.
LABELLING
The label states the K-value.
254
C. vinyl acetate.
Reference: PA/PH/Exp. 13H/T (05) 36 ANP

NOTE ON THE GENERAL METHOD
The method is revised to include a description of the
automated drop point method. This is based on data
obtained from manufacturers of the automated apparatus,
which is more commonly used than the manual one.
XXXX:20217
2.2.17. DROP POINT

The drop point is the temperature at which the first drop of
the melting substance to be examined falls from a cup under
defined conditions.
METHOD A
Apparatus. The apparatus (see Figure 2.2.17.-1) consists
of 2 metal sheaths (A) and (B) screwed together. Sheath
(A) is fixed to a mercury thermometer. A metal cup (F)
is loosely fixed to the lower part of sheath (B) by means
of 2 tightening bands (E). Fixed supports (D) 2 mm long
determine the exact position of the cup, and in addition
are used to centre the thermometer. A hole (C) pierced in
the wall of sheath (B) is used to balance the pressure. The
draining surface of the cup must be flat and the edges of the
outflow orifice must be at right angles to it. The lower part
of the mercury thermometer has the form and size shown
in the figure ; it covers a range from 0 C to 110 C and on
its scale a distance of 1 mm represents a difference of 1 C.
The mercury reservoir of the thermometer has a diameter of
3.5 0.2 mm and a height of 6.0 0.3 mm. The apparatus
is placed in the axis of a test-tube about 200 mm long and
with an external diameter of about 40 mm. It is fixed to the
test-tube by means of a laterally grooved stopper through
which the thermometer passes. The opening of the cup is
placed about 15 mm from the bottom of the test-tube. The
whole device is immersed in a beaker with a capacity of about
1 litre, filled with water. The bottom of the test-tube is placed
about 25 mm from the bottom of the beaker. The water level
reaches the upper part of sheath (A). A stirrer is used to
ensure that the temperature of the water remains uniform.
2.2.17. Drop point
Figure 2.2.17.-1. Apparatus for the determination

of drop point
A. sample cup
D. light slit
G. sample
B. heating block
E. cartridge assembly
H. photo-sensor
C. light source
F. heating element
J. collector sleeve
Figure 2.2.17.-2. Automated drop point apparatus
Method. Melt the substance according to the conditions

described in the monograph. Remove the excess substance
Method. Fill the cup to the brim with the substance to be
at the 2 ends of the cup with a spatula. Condition the
examined, without melting it, unless otherwise prescribed.
sample at the temperature and for the time prescribed in
Remove the excess substance at the 2 ends of the cup with
the monograph before making the measurement. Press
a spatula. When sheaths (A) and (B) have been assembled
the cup into the cup holder, and then press the collector
press the cup into its housing in sheath (B) until it touches sleeve onto the cup. Place the cartridge assembly in the
the supports. Remove with a spatula the substance pushed heating block. Set the instrument to the initial isothermal
out by the thermometer. Place the apparatus in the
conditions and rate for subsequent heating as described in
water-bath as described above. Heat the water-bath and
the monograph of the substance to be examined. Start the
when the temperature is at about 10 C below the presumed temperature programme. When the first drop of molten
drop point, adjust the heating rate to about 1 C/min. Note sample falls through the hole at the bottom of the sample
the temperature at the fall of the first drop. Carry out at
cup, interrupting the light beam, the signal from the
least 3 determinations, each time with a fresh sample of the photo-sensor causes the temperature of the heating block
substance. The difference between the readings must not
to be recorded automatically.
exceed 3 C. The mean of three readings is the drop point
Calibration. Periodically the temperature scale of the
of the substance.
instrument needs to be checked by measuring the drop
point of calibration substances. Pure benzoic acid
and benzophenone are used as these substances melt
METHOD B - AUTOMATED METHOD
without decomposition and have melting points that are
Apparatus. The apparatus (see Figure 2.2.17.-2) consists of
known precisely. They also have little tendency towards
a cartridge assembly comprising a cup holder into which
polymorphism. Prepare 3 sample cups for each of the 2
the sample cup containing the sample is loosely fixed, and
calibration substances benzophenone and benzoic acid.
a collector sleeve with a horizontal light slit, which is fixed
Benzophenone must be pre-dried in a desiccator overnight.
below the cup. This assembly is placed in a heating block.
Place the sample cups on a clean surface. Into each sample
The block is a metal cylinder with a cylindrical hole along
cup, introduce a small quantity of the sample and press it
its vertical axis in which the cartridge assembly is placed.
down with a rod (diameter about 4.5 mm). Check that the
There is another, narrower cylindrical vertical hole in which opening is completely filled. Fill the sample cup about half
a temperature sensor sits. This is positioned level with
full and compact the sample with a rod (diameter about
the sample cup. The heating block is surrounded by an
9 mm). Fill the sample cup completely and compact, adding
electrical heating element. Below the heating block a lamp is more sample and compacting again if necessary, until the
mounted such that a beam of light shines through the light sample cup is completely full.
slit in the collector sleeve, and onto a photo-sensor mounted
opposite. The heating block is capable of being maintained Temperature programme for benzoic acid : start
at a precise, pre-defined temperature by the heating element, temperature = 118.0 C ; heating rate = 0.2 C/min ; end
and of being heated at a slow and steady, pre-defined rate
temperature = 126.0 C. After inserting the cup at 118 C, a
after an initial isothermal period.
waiting time of 30 s is set before heating starts.
Dimensions in millimetres
255
Estradiol benzoate
Temperature programme for benzophenone : start

temperature = 44.0 C ; heating rate = 0.2 C/min ; end
temperature = 56.0 C. After inserting the cup at 44 C, a
waiting time of 30 s is set before heating starts.
Check the 3 single results : none varies by more than 0.3 C
from the mean value. If the differences are greater, prepare
and measure 3 more samples.
Calculate the mean corrected temperature T2 using the
T1
mean drop point temperature of 3 samples, in C ;
correction factor, applied to compensate for the

difference in temperature between the sample
and the point in the heating block where the
temperature is measured ; this will vary depending
upon the design of the automatic drop point
instrument.
Taking into account the melting point (T0) of the calibration

substance, the accuracy of the temperature scale is
satisfactory if |T2 T0| is not greater than 0.3 C.

The test for related substances has been updated in light of
new data received from a manufacturer : a new specified
impurity (impurity G) has been added and a correction
factor for impurity A introduced.
XXXX:0139
ESTRADIOL BENZOATE
Estradioli benzoas
IDENTIFICATION
Comparison : estradiol benzoate CRS.
substance separately in acetone R, evaporate to dryness and
record new spectra using the residues.
TESTS
Specific optical rotation (2.2.7) : + 55.0 to + 59.0 (dried
substance).
Dissolve 0.250 g in acetone R and dilute to 25.0 ml with the
same solvent.
examined in acetonitrile R and dilute to 10.0 ml with the
same solvent.
Reference solution (a). Dissolve 5 mg of estradiol
benzoate impurity E CRS estradiol benzoate for system
suitability CRS (containing impurities A, B, C, E and G) in
5 ml of acetonitrile R , add 2.5 ml of the test solution and
dilute to 10 2.5 ml with acetonitrile R the same solvent.
Reference solution (b). Dilute 1.0 0.5 ml of the test solution
to 100.0 ml with acetonitrile R.
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : octylsilyl silica gel for
Mobile phase :
mobile phase A : water R, acetonitrile R (40:60 V/V) ;
mobile phase B : acetonitrile R ;
Time
(min)
0 - tR 20
Mobile phase A
(per cent V/V)
100
Mobile phase B
(per cent V/V)
0
tR - (tR + 1) 20 - 21
100 10
0 90
(tR + 1) - (tR + 10) 21 - 31
10
90
31 - 32
10 100
90 0
tR = retention time of impurity E

Injection : 10 l.
supplied with estradiol benzoate for system suitability CRS
and the chromatogram obtained with reference solution (a)
to identify the peaks due to impurities A, B, C, E and G.
Elution order : impurity A, impurity F, estradiol benzoate,
C25H28O3
Mr 376.5
impurity E, impurity B, impurity D, impurity C.
Relative retention with reference to estradiol benzoate
DEFINITION
(retention time = about 19 min) : impurity A = about 0.3 ;
17-Hydroxyestra-1,3,5(10)-trien-3-yl benzoate.
impurity E = about 1.1 ; impurity B = about 1.2 ;
impurity G = about 1.3 ; impurity C = about 1.5.
CHARACTERS
Appearance : almost white, crystalline powder or colourless
estradiol benzoate and impurity E.
crystals.
peak-to-valley ratio : minimum 2.0, where Hp = height
Solubility : practically insoluble in water, freely soluble in
above the baseline of the peak due to impurity E and
methylene chloride, sparingly soluble in acetone, slightly
Hv = height above the baseline of the lowest point of the
soluble in methanol.
curve separating this peak from the peak due to estradiol
benzoate.
(10) Zorbax C8 or Kromasil C8 are suitable.
256
Estradiol benzoate
1. impurity A
4. impurity E
7. impurity G
2. impurity F
5. impurity B
8. blank
3. estradiol benzoate
6. impurity D
9. impurity C
Figure 0139.-1. Chromatogram for the test for related substances of estradiol benzoate : test solution spiked with
impurities A to G
Limits :
impurity C = 0.7 ;
impurity C : not more than the area of the principal peak
(0.5 per cent) ;
impurities B, E, G : for each impurity, not more
(0.3 per cent) ;
impurity A : not more than 0.4 times the area of the
any impurity : for each impurity, not more than 0.5 times
(0.10 per cent) ;
total: not more than 1.5 twice the area of the principal
solution (b) (1.5 1.0 per cent) ;
disregard limit : 0.05 0.1 times the area of the principal
solution (b) (0.05 per cent).
Calculate the content of C25H28O3 taking the specific

absorbance to be 500.
IMPURITIES
Specified impurities : A, B, C, D, E, F, G.
pharmaceutical use) : D, F.
A. R1 = R2 = R3 = H, R4 = OH : estradiol,
B. R1 = CO-C6H5, R2 = CH3, R3 = H, R4 = OH :
17-hydroxy-4-methylestra-1,3,5(10)-trien-3-yl benzoate,
C. R1 = CO-C6H5, R2 = R3 = H, R4 = O-CO-C6H5 :
estra-1,3,5(10)-triene-3,17-diyl dibenzoate,
ASSAY
D. R1 = R2 = R3 = H, R4 = O-CO-C6H5 : 3-hydroxyestra-1,3,
5(10)-trien-17-yl benzoate,
Dissolve 25.0 mg in anhydrous ethanol R and dilute to
250.0 ml with the same solvent. Dilute 10.0 ml of this
solution to 100.0 ml with anhydrous ethanol R. Measure the E. R1 = CO-C6H5, R2 = R4 = H, R3 = OH :
absorbance (2.2.25) at the absorption maximum at 231 nm.
17-hydroxyestra-1,3,5(10)-trien-3-yl benzoate,
257
Eye preparations
F. 17-hydroxyestra-1,3,5(10),9(11)-tetraen-3-yl benzoate,
G. 17-oxo-estra-1,3,5(10)-trien-3-yl benzoate.
PRODUCTION
During the development of an eye preparation, whose
formulation contains an antimicrobial preservative, the
necessity for and the efficacy of the chosen preservative
shall be demonstrated to the satisfaction of the competent
authority. A suitable test method together with criteria
for judging the preservative properties of the formulation
are provided in the text on Efficacy of antimicrobial
preservation (5.1.3).
Eye preparations are prepared using materials and methods
designed to ensure sterility and to avoid the introduction
of contaminants and the growth of micro-organisms ;
recommendations on this aspect are provided in the text on
Methods of preparation of sterile products (5.1.1).
In the manufacture of eye preparations containing dispersed
particles, measures are taken to ensure a suitable and
controlled particle size with regard to the intended use.
During development, it must be demonstrated that the
nominal content can be withdrawn from the container of
liquid and semi-solid eye preparations supplied in single-dose
containers.
TESTS
Sterility (2.6.1). Eye preparations comply with the test for
sterility. Applicators supplied separately also comply with
the test for sterility. Remove the applicator with aseptic
A systematic revision of the monographs prescribing the
precautions from its package and transfer it to a tube of
test for Deliverable mass or volume is being carried out,
culture medium so that it is completely immersed. Incubate
with a view to deleting the test because of its weakness (only and interpret the results as described in the test for sterility.
one container is tested). Instead, a general requirement is
Deliverable mass or volume (2.9.28). Liquid and semi-solid
added under Production. This corresponds to a European
eye preparations supplied in single-dose containers comply
legal requirement for trade.
with the test.
Eye drops : the possibility to use multidose containers for
eye drops that do not contain antimicrobial preservatives
STORAGE
is now offered, provided these multidose containers are
Unless otherwise prescribed, store in a sterile, airtight,
designed to avoid microbial contamination.
tamper-proof container.
Semi-solid eye preparations : in the case of eye lotions and
LABELLING
eye drops, a larger container than that prescribed in the
monograph may be used, where justified and authorised ;
The label states the name of any added antimicrobial
it is proposed to extend this possibility to semi-solid eye
preservative.
preparations, as it appears that preparations in containers
containing more than 10 g would already be marketed
Eye drops
in Europe. The readers are invited to comment on this
extension, in particular in view of the risk of microbial
DEFINITION
contamination.
Eye drops are sterile aqueous or oily solutions, emulsions or
XXXX:1163 suspensions of one or more active substances intended for
instillation into the eye.
EYE PREPARATIONS
Eye drops may contain excipients, for example, to adjust
the tonicity or the viscosity of the preparation, to adjust
Ophthalmica
or stabilise the pH, to increase the solubility of the active
substance, or to stabilise the preparation. These substances
DEFINITION
do not adversely affect the intended medicinal action or, at
Eye preparations are sterile liquid, semi-solid or solid
the concentrations used, cause undue local irritation.
preparations intended for administration upon the eyeball
Aqueous preparations supplied in multidose containers
and/or to the conjunctiva, or for insertion in the conjunctival contain a suitable antimicrobial preservative in appropriate
sac.
concentration except when the preparation itself has
Where applicable, containers for eye preparations comply
adequate antimicrobial properties. The antimicrobial
with the requirements of Materials used for the manufacture preservative chosen must be compatible with the other
of containers (3.1 and subsections) and Containers (3.2 and ingredients of the preparation and must remain effective
subsections).
throughout the period of time during which eye drops are
Several categories of eye preparations may be distinguished : in use.
If eye drops are prescribed without do not contain
eye drops ;
antimicrobial preservatives they are supplied wherever
eye lotions ;
possible in single-dose containers or in multidose containers
powders for eye drops and powders for eye lotions ;
designed to deliver the preparation without microbial
semi-solid eye preparations ;
contamination. Eye drops intended for use in surgical
ophthalmic inserts.
procedures do not contain antimicrobial preservatives ; they
258
Eye preparations
and are supplied in single-dose containers or in multidose

containers designed to deliver the preparation without
microbial contamination.
Eye drops that are solutions, examined under suitable
conditions of visibility, are practically clear and practically
free from particles.
Eye drops that are suspensions may show a sediment that is
readily redispersed on shaking to give a suspension which
remains sufficiently stable to enable the correct dose to be
delivered.
Multidose preparations are supplied in containers that allow
successive drops of the preparation to be administered. The
containers contain at most 10 ml of the preparation, unless
otherwise justified and authorised.
TESTS
Particle size. Unless otherwise justified and authorised, eye
drops in the form of a suspension comply with the following
test : introduce a suitable quantity of the suspension
into a counting cell or with a micropipette onto a slide,
as appropriate, and scan under a microscope an area
corresponding to 10 g of the solid phase. For practical
reasons, it is recommended that the whole sample is first
scanned at low magnification (e.g. 50) and particles greater
than 25 m are identified. These larger particles can then
be measured at a larger magnification (e.g. 200 to 500).
For each 10 g of solid active substance, not more than 20
particles have a maximum dimension greater than 25 m,
and not more than 2 of these particles have a maximum
dimension greater than 50 m. None of the particles has a
maximum dimension greater than 90 m.
LABELLING
The label states :
where applicable, that the contents are to be used on one
occasion only ;
for multidose containers, the period after opening the
container after which the contents must not be used.
This period does not exceed 4 weeks, unless otherwise
justified and authorised.
Powders for eye drops and powders for eye

lotions
DEFINITION
Powders for the preparation of eye drops and eye lotions are
supplied in a dry, sterile form to be dissolved or suspended
in an appropriate liquid vehicle at the time of administration.
They may contain excipients to facilitate dissolution or
dispersion, to prevent caking, to adjust the tonicity, to adjust
or stabilise the pH or to stabilise the preparation.
After dissolution or suspension in the prescribed liquid, they
comply with the requirements for eye drops or eye lotions, as
appropriate.
TESTS
Uniformity of dosage units. Single-dose powders for eye
drops and eye lotions comply with the test for uniformity of
dosage units (2.9.40) or, where justified and authorised, with
the tests for uniformity of content and/or uniformity of mass
shown below. Herbal drugs and herbal drug preparations
present in the dosage form are not subject to the provisions
of this paragraph.
LABELLING
Uniformity of content (2.9.6). Unless otherwise prescribed
or justified and authorised, single-dose powders for eye
The label states, for multidose containers, the period after
opening the container after which the contents must not be drops and eye lotions with a content of active substance
used. This period does not exceed 4 weeks, unless otherwise less then 2 mg or less than 2 per cent of the total mass
comply with test B for uniformity of content of single-dose
preparations. If the preparation has more than one active
substance, the requirement applies only to those substances
Eye lotions
that correspond to the above condition.
Uniformity of mass (2.9.5). Single-dose powders for eye
DEFINITION
Eye lotions are sterile aqueous solutions intended for use in drops and eye lotions comply with the test for uniformity of
rinsing or bathing the eye or for impregnating eye dressings. mass of single-dose preparations. If the test for uniformity of
content is prescribed for all the active substances, the test
Eye lotions may contain excipients, for example to adjust
for uniformity of mass is not required.
the tonicity or the viscosity of the preparation or to adjust
or stabilise the pH. These substances do not adversely affect
Semi-solid eye preparations
the intended action or, at the concentrations used, cause
undue local irritation.
DEFINITION
Eye lotions supplied in multidose containers contain
Semi-solid eye preparations are sterile ointments, creams
a suitable antimicrobial preservative in appropriate
or gels intended for application to the conjunctiva. They
concentration except when the preparation itself has
contain one or more active substances dissolved or dispersed
adequate antimicrobial properties. The antimicrobial
in a suitable basis. They have a homogeneous appearance.
preservative chosen is compatible with the other ingredients
Semi-solid eye preparations comply with the requirements of
of the preparation and remains effective throughout the
the monograph on Semi-solid preparations for cutaneous
period of time during which the eye lotions are in use.
application (0132). The basis is non-irritant to the
If eye lotions are prescribed without an do not contain
conjunctiva.
antimicrobial preservatives, they are supplied in single-dose
Semi-solid eye preparations are packed in small, sterilised
containers. Eye lotions intended for use in surgical
collapsible tubes fitted or provided with a sterilised
procedures or in first-aid treatment do not contain an
cannula and having a content of not more than. The
antimicrobial preservative and are supplied in single-dose
containers contain at most 10 g of the preparation, unless
containers.
otherwise justified and authorised. The tubes must be
Eye lotions, examined under suitable conditions of visibility, well-closed to prevent microbial contamination. Semi-solid
are practically clear and practically free from particles.
eye preparations may also be packed in suitably designed
single-dose containers. The containers, or the nozzles of
The containers for multidose preparations do not contain
tubes, are of such a shape as to facilitate administration
more than 200 ml of eye lotion, unless otherwise justified
without contamination.
and authorised.
259
Reference: PA/PH/Exp. 10A/T (03) 16 ANP 1R
TESTS
Particle size. Semi-solid eye preparations containing
dispersed solid particles comply with the following test :
spread gently a quantity of the preparation corresponding
to at least 10 g of solid active substance as a thin layer.
Scan under a microscope the whole area of the sample. For
practical reasons, it is recommended that the whole sample is
first scanned at a small magnification (e.g. 50) and particles
greater than 25 m are identified. These larger particles
can then be measured at a larger magnification (e.g. 200
to 500). For each 10 g of solid active substance, not
more than 20 particles have a maximum dimension greater
than 25 m, and not more than 2 of these particles have
a maximum dimension greater than 50 m. None of the
particles has a maximum dimension greater than 90 m.

A monograph proposal including a test for related
substances by TLC was proposed in Pharmeuropa 15.3.
An LC method is now available and this new proposal is
presented.
XXXX:1692
FLAVOXATE HYDROCHLORIDE
Flavoxati hydrochloridum
LABELLING
The label states, for multidose containers, the period after
opening the container after which the contents must not be
used. This period does not exceed 4 weeks, unless otherwise
C24H26ClNO4
Mr 427.9
Ophthalmic inserts
DEFINITION
2-(Piperidin-1-yl)ethyl 3-methyl-4-oxo-2-phenyl-4H-1benzopyran-8-carboxylate hydrochloride.
Ophthalmic inserts are sterile, solid or semi-solid

preparations of suitable size and shape, designed to be
inserted in the conjunctival sac, to produce an ocular effect.
They generally consist of a reservoir of active substance
embedded in a matrix or bounded by a rate-controlling
membrane. The active substance, which is more or less
soluble in physiological fluids, is released over a determined
period of time.
CHARACTERS
Solubility : slightly soluble in water, sparingly soluble in
methylene chloride, slightly soluble in ethanol (96 per cent).
DEFINITION
Ophthalmic inserts are individually distributed into sterile

containers.
PRODUCTION
In the manufacture of ophthalmic inserts, measures are
taken to ensure a suitable dissolution behaviour.
TESTS
Uniformity of dosage units. Ophthalmic inserts comply
with the test for uniformity of dosage units (2.9.40) or,
where justified and authorised, with the test for uniformity
of content shown below. Herbal drugs and herbal drug
preparations present in the dosage form are not subject to
the provisions of this paragraph.
Uniformity of content (2.9.6). Ophthalmic inserts comply,
where applicable, with test A for uniformity of content.
LABELLING
The label states :
where applicable, the total quantity of active substance
per insert ;
where applicable, the dose released per unit time.
IDENTIFICATION
Comparison : flavoxate hydrochloride CRS.
B. It gives reaction (a) of chlorides (2.3.1).
TESTS
Related substances. Liquid chromatography (2.2.29). Use
freshly prepared solutions.
Solvent mixture. Mix 20 volumes of a 0.408 g/l solution of
potassium dihydrogen phosphate R adjusted to pH 3.0 with
phosphoric acid R and 80 volumes of acetonitrile R.
examined in the solvent mixture and dilute to 10.0 ml with
the solvent mixture.
100.0 ml with the solvent mixture.
to 10.0 ml with the solvent mixture.
Reference solution (c). Dissolve 6.0 mg of flavoxate
impurity A CRS and 3.0 mg of flavoxate impurity B CRS
in the solvent mixture, add 2.0 ml of the test solution and
dilute to 100.0 ml with the solvent mixture. Dilute 1.0 ml of
this solution to 20.0 ml with the solvent mixture.
Column :
size : l = 0.25 m, = 4.6 mm ;
(11) Hypersil BDS, Discovery C18, Kromasil C18 are suitable.
260
2. impurity B
1. impurity A
3. impurity C
4. flavoxate
Figure 1692.-1. Chromatogram for the test for related substances of flavoxate hydrochloride
Mobile phase :
mobile phase A : 0.435 g/l solution of dipotassium
hydrogen phosphate R adjusted to pH 7.5 with
phosphoric acid R ;
Time
(min)
0 - 10
Mobile phase A
(per cent V/V)
20
Mobile phase B
(per cent V/V)
80
10 - 20
20 10
80 90
20 - 25
10
90
25 - 27
10 20
90 80
27 - 35
20
80

Equilibration: at least 30 min with the mobile phase at the
initial composition.
Injection: 10 l.
Relative retention with reference to flavoxate (retention
System suitability : reference solution (c) :
impurity B and flavoxate.
Limits :
solution (c) (0.3 per cent) ;
impurity B : not more than the area of the corresponding
solution (c) (0.15 per cent) ;
total : not more than the area of the principal peak in

the chromatogram obtained with reference solution (a)
(1.0 per cent) ;
(0.05 per cent).
on 1.000 g by drying in an oven at 105 C.
on 1.0 g.
ASSAY
In order to avoid overheating in the reaction medium, mix
thoroughly throughout and stop the titration immediately
after the end-point has been reached.
Dissolve 0.350 g in 10 ml of anhydrous formic acid R and add
40 ml of acetic anhydride R. Titrate with 0.1 M perchloric
acid, determining the end-point potentiometrically (2.2.20).
C24H26ClNO4.
STORAGE
Protected from light.
IMPURITIES
Specified impurities : A, B.
pharmaceutical use) : C.
261
2.8.20. Herbal drugs : sampling and sample preparation
A. R = H : 3-methyl-4-oxo-2-phenyl-4H-1-benzopyran-8carboxylic acid,
B. R = C2H5 : ethyl 3-methyl-4-oxo-2-phenyl-4H-1-benzopyran8-carboxylate,
C. R = CH(CH3)2 : isopropyl 3-methyl-4-oxo-2-phenyl-4H-1benzopyran-8-carboxylate.
Reference: PA/PH/Exp. 13B/T (04) 10 ANP 1R

NOTE ON THE GENERAL METHOD
This draft monograph, previously published in
Pharmeuropa 16.4, has been substantially modified to
take account of the comments from national authorities.
However, there remain two alternative approaches to the
number of containers to be sampled. The two approaches
are given as option 1 and option 2 with an illustrative guide
given in Tables 2.8.20.-1 and 2.8.20.-2 as to the number of
containers to be sampled and the weight of bulk sample
required for each approach. Comments are requested on
the suitability of these two approaches for the sampling
of herbal drugs.
XXXX:20820
2.8.20. HERBAL DRUGS : SAMPLING

AND SAMPLE PREPARATION
In order to reduce the effect of sampling in qualitative and
quantitative analysis, it is necessary to ensure that the
composition of the sample used is representative of the batch
of material being examined. The following procedures are
the minimum considered applicable for herbal drugs. NOTE :
other procedures may be used if they can be demonstrated
to produce representative batch samples.
BULK SAMPLE
Where external examination of containers, markings and
labels of a batch indicates that it can be considered to be
homogeneous, sample the number of randomly selected
containers indicated below. Where a batch cannot be
considered to be homogeneous, divide it into sub-batches
that are as homogeneous as possible, then sample each
sub-batch as a homogeneous batch according to the number
of randomly selected containers indicated below.
Number of containers in batch (N)
Number of containers
to be sampled (n)
1 - 12
Option 1
Option 2
> 12
n* =
1-3
>3
* round n up to the next integer
262
all
all
n* =
Take one sample from each container to be sampled. The

sample is taken from the upper, middle or lower section of
the container, such that the samples taken are representative
of different parts of the containers. In the case of large bales
or bags, samples must be taken from a depth of at least
10 cm. The weight of the material taken from each container
is such that the total weight of the bulk sample complies
with the following values.
Weight of herbal drug in the
batch (kg)
Total weight of samples as a

percentage of the weight of the
batch of herbal drug
< 50
1.00*
50 - 100
0.50
> 100 - 250
0.25
> 250 - 500
0.20
> 500 - 1000
0.18
> 1000 - 2500
0.15
> 2500 - 5000
0.10
> 5000 - 10 000
0.08
> 10 000 - 25 000
0.05
NOTE : if the weight of the batch is greater than 25 000 kg, it is divided
into sub-batches, and the procedure is applied to each sub-batch as
though it were a homogeneous batch.
* subject to a minimum total weight of 125 g for the bulk sample ; if this
minimum requirement represents more than 10.0 per cent of the weight
of herbal drug in the batch, the whole batch may be used as the sample.
Prepare the bulk sample by combining and thoroughly

mixing the samples taken from each of the randomly selected
containers.
TEST SAMPLE
Unless otherwise prescribed in the monograph, prepare the
test sample as follows.
Reduce the size of the bulk sample by quartering (see Note
below) or by any other method that produces a homogeneous
sample, making sure that each retained portion remains
representative of the whole, until the minimum retained
quantity complies with the following conditions.
Type of herbal drug
Minimum weight of test sample
Roots, rhizomes, bark, herbs
500 g or weight of whole sample if

bulk sample is less than 500 g
Leaves, flowers, seeds, fruits
250 g or weight of whole sample if

bulk sample is less than 250 g
Broken or fragmented drugs

(where average weight of the
pieces is less than 0.5 g)
125 g
NOTE : quartering consists of placing the bulk sample,

thoroughly mixed, as a level and square-shaped heap
and dividing it diagonally into 4 equal parts. 2 opposite
quarters are retained and carefully remixed. The process is
repeated as necessary until the required minimum weight
is obtained for the test sample.
Mill the test sample in a single pass through a 1 mm screen
or the screen size specified in the monograph. The use of a
milling machine is recommended.
Pass the milled sample through a 1 mm standard sieve or
the sieve specified in the monograph. The residue retained
on the sieve must not be more than 10 per cent of the total
weight of the milled sample and not more than 2 per cent
may be of a particle size greater than 1.5 mm or 1.5 times the
specified particle size in the monograph. If these conditions
are met the sample and residue are to be well mixed to form
the test sample for analysis.
The following table is shown for information but will not be published in the European Pharmacopoeia.
Table 2.8.20.-1. Operation of the sampling procedure in order to obtain the prescribed bulk sample according to option 1
263
The following table is shown for information but will not be published in the European Pharmacopoeia.
Table 2.8.20.-2. Operation of the sampling procedure in order to obtain the prescribed bulk sample according to option 2
264
Dissolve half of the precipitate in 1 ml of ethanol (96 per

In those cases where these requirements are not met the
cent) R and add 0.5 ml of a 100 g/l solution of cobalt
test sample for analysis is composed of the 2 parts measured
nitrate R. A bluish-green precipitate is formed.
separately. Therefore the quantity required for each analysis
is derived by weighing proportional quantities of the powder F. It gives reaction (a) of chlorides (2.3.1).
and residue.
NOTE : for determination of microscopic characters, a
TESTS
portion of the milled test sample is re-milled though a
0.355 mm screen.
and dilute to 20 ml with the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and
colourless (2.2.2, Method II).
pH (2.2.3) : 4.0 to 5.5.
Reference: PA/PH/Exp. 10A/T (06) 5 ANP
Dilute 1 ml of solution S to 10 ml with carbon dioxide-free
water R.
Impurity A
The revision includes the replacement of the TLC for
impurity A by an LC, which allows the control of impurity A Solution (a). Dissolve 0.25 g of the substance to be examined
in methanol R and dilute to 10 ml with the same solvent.
and other related substances.
XXXX:0227 This solution is used to prepare the test solution.
Solution (b). Dissolve 50 mg of 2,6-dimethylaniline R in
methanol R and dilute to 100 ml with the same solvent.
LIDOCAINE HYDROCHLORIDE
Dilute 1 ml of the solution to 100 ml with methanol R. This
solution is used to prepare the standard.
Lidocaini hydrochloridum
Using three flat-bottomed tubes, place in the first 2 ml of
solution (a), in the second 1 ml of solution (b) and 1 ml
of methanol R and in the third 2 ml of methanol R (used
to prepare a blank). To each tube add 1 ml of a freshly
prepared 10 g/l solution of dimethylaminobenzaldehyde R
in methanol R and 2 ml of glacial acetic acid R and allow
to stand at room temperature for 10 min. The intensity of
the yellow colour of the test solution is between that of the
C14H23ClN2O,H2O
Mr 288.8 blank and that of the standard (100 ppm).
DEFINITION
2-(Diethylamino)-N-(2,6-dimethylphenyl)acetamide
examined in the mobile phase and dilute to 10.0 ml with the
hydrochloride monohydrate.
mobile phase.
Content : 99.0 per cent to 101.0 per cent (anhydrous
Reference solution (a). Dissolve 10.0 mg of lidocaine
substance).
impurity A CRS in the mobile phase and dilute to 200.0 ml
with the mobile phase.
CHARACTERS
Reference solution (b). Dissolve 5 mg of
2-chloro-N-(2,6-dimethylphenyl)acetamide R (impurity H) in
Solubility : very soluble in water, freely soluble in ethanol
the mobile phase and dilute to 10.0 ml with the mobile phase.
(96 per cent).
Reference solution (c). Dilute 1.0 ml of the test solution to
IDENTIFICATION
10.0 ml with the mobile phase.
First identification : A, B, F.
Reference solution (d). Mix 1.0 ml of reference solution (a),
1.0 ml of reference solution (b) and 1.0 ml of reference
Second identification : A, C, D, E, F.
solution (c) and dilute to 100.0 ml with the mobile phase.
A. Melting point (2.2.14) : 74 C to 79 C, determined
Column :
without previous drying.
size : l = 0.15 m, = 3.9 mm ;
Comparison : lidocaine hydrochloride CRS.
stationary phase : end-capped polar-embedded
octadecylsilyl amorphous organosilica polymer R
C. Dissolve 0.2 g in 10 ml of water R and add 10 ml of picric
(5 m)(12) ;
acid solution R. The precipitate, washed with water R
and dried, melts (2.2.14) at about 230 C.
temperature : 30 C.
D. To about 5 mg add 0.5 ml of fuming nitric acid R.
Mobile phase : mix 30 volumes of acetonitrile for
Evaporate to dryness on a water-bath, cool and dissolve
chromatography R and 70 volumes of a 4.85 g/l solution of
the residue in 5 ml of acetone R. Add 0.2 ml of alcoholic potassium dihydrogen phosphate R adjusted to pH 8.0 with
potassium hydroxide solution R. A green colour is
strong sodium hydroxide solution R.
produced.
E. To 5 ml of solution S (see Tests) add 5 ml of water R and
make alkaline with dilute sodium hydroxide solution R. Detection : spectrophotometer at 230 nm.
Collect the precipitate on a filter and wash with water R. Injection : 20 l(13).
(12) XTerra RP C18 is suitable.
(13) A slight carry-over might occasionally be observed ; this can be resolved by rinsing the injector.
265
1. impurity C
3. impurity G
5. impurity A
7. lidocaine
9. impurity I
2. impurity D
4. impurity H
6. impurity E
8. impurity F
10. impurity J
Figure 0227.-1. Chromatogram for the test for related substances of lidocaine hydrochloride
Relative retention with reference to lidocaine (retention
time = about 17 min) : impurity H = about 0.37 ;
System suitability : reference solution (d) :
Water (2.5.12) : 5.5 per cent to 7.0 per cent, determined on

0.25 g.
on 1.0 g.

impurities A and H.
ASSAY
Dissolve 0.220 g in 50 ml of ethanol (96 per cent) R
and add 5.0 ml of 0.01 M hydrochloric acid. Carry out
Limits :
a potentiometric titration (2.2.20), using 0.1 M sodium
impurity A : not more than the area of the corresponding hydroxide. Read the volume added between the 2 points
of inflexion.
solution (d) (0.01 per cent) ;
1 ml of 0.1 M sodium hydroxide is equivalent to 27.08 mg
unspecified impurities : for each impurity, not more than of C14H23ClN2O.
the area of the peak due to lidocaine in the chromatogram
STORAGE
obtained with reference solution (d) (0.10 per cent) ;
total: not more than 5 times the area of the peak due to
lidocaine in the chromatogram obtained with reference
IMPURITIES
solution (d) (0.5 per cent) ;
Specified impurities : A.
disregard limit : 0.5 times the area of the peak due to
lidocaine in the chromatogram obtained with reference
solution (d) (0.05 per cent).
Dissolve 1.0 g in water R and dilute to 25 ml with the same pharmaceutical use (2034). It is therefore not necessary to
solvent. Carry out the prefiltration. 10 ml of the prefiltrate
complies with test E. Prepare the reference solution using
2 ml of lead standard solution (1 ppm Pb) R.
pharmaceutical use) : B, C, D, E, F, G, H, I, J.
266
A. 2,6-dimethylaniline.
J. 2-(diethylamino)-N-(2,5-dimethylphenyl)acetamide.
Reagents
2-Chloro-N-(2,6-dimethylphenyl)acetamide. C10H12ClNO.
(Mr 197.7). XXXXXXX. [1131-01-7].
B. 2-(diethylnitroryl)-N-(2,6-dimethylphenyl)acetamide
(lidocaine N-oxyde),
C. N-(2,6-dimethylphenyl)acetamide,
D. N-(2,6-dimethylphenyl)-2-(ethylamino)acetamide,

The degree of hydration is now given in the title to clarify
the scope of the monograph. Lincomycin B, previously
described as an impurity, has been integrated into the
definition of the substance, since it has been proven to
participate fully in the antimicrobial activity. The second
identification series has been deleted since it has no
practical application. The GC assay has been replaced by
a less labour-intensive, more sensitive and more selective
LC assay.
XXXX:0583
LINCOMYCIN HYDROCHLORIDE
MONOHYDRATE
E. 2,2-iminobis(N-(2,6-dimethylphenyl)acetamide),
Lincomycini hydrochloridum
monohydricum
F. 2-(diethylamino)-N-(2,3-dimethylphenyl)acetamide,
G. N-(2,6-dimethylphenyl)-2-((1-methylethyl)amino)acetamide,
H. 2-chloro-N-(2,6-dimethylphenyl)acetamide,
I. 2-(diethylamino)-N-(2,4-dimethylphenyl)acetamide,
C18H35ClN2O6S,H2O
Mr 461.0
DEFINITION
Lincomycin hydrochloride consists mainly of Mixture of
antibiotics, the main component being methyl 6,8-dideoxy-6[[[(2S,4R)-1-methyl-4-propylpyrrolidin-2-yl]carbonyl]amino]1-thio-D-erythro--D-galacto-octopyranoside hydrochloride
monohydrate (lincomycin hydrochloride), an antimicrobial
substance produced by Streptomyces lincolnensis var.
lincolnensis or by any other means.
Content :
It contains not less than 89.5 per cent and not more
than 102.0 per cent of lincomycin hydrochloride
(C18H34N2O6S.HCl), calculated with reference to the
anhydrous substance.
267
sum of the contents of lincomycin and lincomycin B :

89.5 96.0 per cent to 102.0 per cent (anhydrous
substance) ;
lincomycin B : maximum 5.0 per cent.
CHARACTERS
Solubility : very soluble in water, slightly soluble in ethanol
(96 per cent), very slightly soluble in acetone.
IDENTIFICATION
First identification : A, D.
Second identification : B, C, D.
Comparison : lincomycin hydrochloride CRS.
B. Thin-layer chromatography (2.2.27).
examined in methanol R and dilute to 10 ml with the
same solvent.
Reference solution (a). Dissolve 10 mg of lincomycin
hydrochloride CRS in methanol R and dilute to 10 ml
with the same solvent.
Reference solution (b). Dissolve 10 mg of lincomycin
hydrochloride CRS and 10 mg of clindamycin
hydrochloride CRS in methanol R and dilute to 10 ml
Plate : silica gel G R.
Mobile phase : mix 20 volumes of 2-propanol R,
40 volumes of a 150 g/l solution of ammonium acetate R
previously adjusted to pH 9.6 with ammonia R and
45 volumes of ethyl acetate R.
Application : 5 l.
Detection : spray with a 1 g/l solution of potassium
permanganate R.
Drying : in air.
Results : the principal spot in the chromatogram obtained
with the test solution is similar in position, colour and
size to the principal spot in the chromatogram obtained
with reference solution (a). The test is not valid unless
the chromatogram obtained with reference solution (b)
shows two clearly separated spots.
C. Dissolve about 10 mg in 2 ml of dilute hydrochloric acid R
and heat in a water-bath for 3 min. Add 3 ml of sodium
carbonate solution R and 1 ml of a 20 g/l solution of
sodium nitroprusside R. A violet-red colour develops.
D. B. Dissolve 0.1 g in water R and dilute to 10 ml with the
same solvent. The solution gives reaction (a) of chlorides
(2.3.1).
TESTS
and dilute to 20 ml with the same solvent.
Appearance of solution. Solution S is clear (2.2.1) and not
more intensely coloured than reference solution Y6 (2.2.2,
Method II).
Specific optical rotation (2.2.7) : + 135 to + 150 (anhydrous
substance).
Dissolve 1.000 g in water R and dilute to 25.0 ml with the
same solvent.
Lincomycin B. Examine the chromatogram obtained in

the assay with test solution (a). The area of the peak due
to lincomycin B, which is eluted just before lincomycin, is
not more than 5 per cent of the area of the peak due to
lincomycin.
solvent.
Reference solution (a). Dissolve 50.0 mg of lincomycin
hydrochloride CRS in water R and dilute to 25.0 ml with
the same solvent.
100.0 ml with water R.
to 50.0 ml with water R.
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : base-deactivated octadecylsilyl silica
gel for chromatography R (5 m)(14) ;
temperature : 45 C.
Mobile phase : mix 50.0 ml of acetonitrile R and 50.0 ml of
a 27.2 g/l solution of potassium dihydrogen phosphate R
adjusted to pH 5.0 with a 34.8 g/l solution of dipotassium
hydrogen phosphate R, add 0.50 g of methanesulphonic
acid R and dilute to 1000 ml with water R.
Injection : 20 l of the test solution and reference
solutions (b) and (c).
Run time : twice the retention time of lincomycin.
Relative retention with reference to lincomycin (retention
time = about 18 min) : lincomycin B = about 0.44.
resolution : minimum 10 between the peaks due to
lincomycin and lincomycin B.
Limits :
any impurity : for each impurity, not more than the area
of the principal peak in the chromatogram obtained with
reference solution (c) (0.2 per cent) ;
total : not more than 0.4 times the area of the principal
solution (b) (2.0 per cent) ;
in the chromatogram obtained with reference solution (c)
(0.1 per cent).
using 1.0 ml of lead standard solution (10 ppm Pb) R.
0.500 g.
on 1.0 g.
Bacterial endotoxins (2.6.14) : less than 0.50 IU/mg, if
intended for use in the manufacture of parenteral dosage
forms without a further appropriate procedure for the
removal of bacterial endotoxins.
ASSAY
Examine by gas chromatography (2.2.28), using
dotriacontane R as the internal standard.
(14) Supelcosil DB LC-18 is suitable.
268
Macrogol 40 sorbitol heptaoleate
Internal standard solution. Dissolve 0.200 g of

dotriacontane R in chloroform R and dilute to 25.0 ml with
the same solvent.
examined in a 20 g/l solution of imidazole R in chloroform R
and dilute to 100.0 ml with the same solution. Shake until
dissolution is complete. Place 4.0 ml of the solution in a
ground-glass-stoppered 15 ml centrifuge tube. Add 1.0 ml
of a mixture of 1 volume of chlorotrimethylsilane R and
A. R1 = CH2-CH2-CH3, R2 = R3 = H, R4 = OH : methyl
99 volumes of N,O-bis(trimethylsilyl)acetamide R and swirl
6,8-dideoxy-6-[[[(2S,4R)-1-methyl-4-propylpyrrolidingently. Position the glass stopper loosely in the tube and
2-yl]carbonyl]amino]-1-thio-L-threo--D-galactoheat at 65 C for 30 min.
octopyranoside (7-epilincomycin).
Test solution (b). Prepare as described for test solution (a)
but add 10.0 ml of the internal standard solution before
dissolution of the substance to be examined.
Reference solution. Prepare as described for test solution (a)
using 0.100 g of lincomycin hydrochloride CRS instead of
the substance to be examined and adding 10.0 ml of the
internal standard solution before dissolution of the reference
substance.
The chromatographic procedure may be carried out using :
a glass column 1.5 m long and 3 mm in internal diameter
packed with silanised diatomaceous earth for gas
chromatography R impregnated with 3 per cent m/m of
poly(methylphenylsiloxane) R,
Reference: PA/PH/Exp. 13H/T (05) 68 ANP

XXXX:2396
MACROGOL 40 SORBITOL
HEPTAOLEATE
Macrogol 40 sorbitoli heptaoleas
helium for chromatography R as the carrier gas at a flow DEFINITION

rate of about 45 ml/min,
Mixture of esters of fatty acids, mainly Oleic acid (0799),
and sorbitol ethoxylated with approximately 40 moles of
a flame-ionisation detector,
ethylene oxide for each mole of sorbitol.
maintaining the temperature of the column at 260 C and
that of the injection port and of the detector between 260 C CHARACTERS
and 290 C. Inject the chosen volume of the test solutions
Appearance : clear or slightly opalescent, yellowish, viscous,
and the reference solution.
hygroscopic liquid.
Liquid chromatography (2.2.29) as described in the test for Solubility : dispersible in water, soluble in isopropyl
related substances with the following modifications.
myristate, in isopropyl palmitate, in mineral oils and in
vegetable fatty oils.
Injection : the test solution and reference solution (a).
Relative density : about 1.0.
Calculate the percentage content of C18H35ClN2O6S and of
C17H33ClN2O6S from the declared content of lincomycin and Viscosity (2.2.9) : about 175 mPas at 25 C.
of lincomycin B in lincomycin hydrochloride CRS.
IDENTIFICATION
First identification : A, D.
STORAGE
Store in an airtight container aAt a temperature not
exceeding 30 C. If the substance is sterile, store in a sterile, A. Infrared absorption spectrophotometry (2.2.24).
airtight, tamper-proof container.
Comparison : macrogol 40 sorbitol heptaoleate CRS.
B. Hydroxyl value (see Tests).
C. Saponification value (see Tests).
LABELLING
The label states, where applicable, that the substance is free D. Composition of fatty acids (see Tests).
from bacterial endotoxins.
TESTS
Acid value (2.5.1) : maximum 12.0, determined on 3.0 g.
IMPURITIES
Hydroxyl value (2.5.3, Method A) : 22 to 38.
Peroxide value : maximum 10.0.
Introduce 10.0 g into a 100 ml beaker, dissolve with glacial
acetic acid R and dilute to 20 ml with the same acid. Add
impurities and/or by the general monograph Substances for 1 ml of saturated potassium iodide solution R and allow to
pharmaceutical use (2034). It is therefore not necessary to stand for 1 min. Add 50 ml of carbon dioxide-free water R
and a magnetic stirring bar. Titrate with 0.01 M sodium
thiosulphate, determining the end-point potentiometrically
pharmaceutical use) : A.
(2.2.20). Carry out a blank titration.
269
Figure 2396.-1. Chromatogram for the test for composition of fatty acids of macrogol 40 sorbitol heptaoleate
Determine the peroxide value using the following expression : Sulphated ash : maximum 0.25 per cent.
Heat a silica crucible to redness for 30 min, allow to cool
in a desiccator and weigh. Evenly distribute 1.00 g of the
substance to be examined in the crucible and weigh. Dry
at 100-105 C for 1 h and ignite in a muffle furnace at
600 C 25 C, until the substance is thoroughly charred.
n1 = volume of 0.01 M sodium thiosulphate required
Carry out the test for sulphated ash (2.4.14) on the residue
for the titration of the substance to be examined,
obtained, starting from Moisten the substance to be
in millilitres;
examined....
n2 = volume of 0.01 M sodium thiosulphate required
STORAGE
for the blank titration, in millilitres;
M
= molarity of the sodium thiosulphate solution, in
moles per litre;
m
= mass of the substance to be examined, in grams.
Saponification value (2.5.6) : 100 to 110, determined on
4.0 g.
XXXX:1788
Use 30.0 ml of 0.5 M alcoholic potassium hydroxide,
heat under reflux for 60 min and add 50 ml of anhydrous
METHYLERGOMETRINE HYDROGEN
ethanol R before carrying out the titration.
MALEATE
Composition of fatty acids (2.4.22, Method C). Use the
mixture of calibrating substances in Table 2.4.22.-3.
Methylergometrini hydrogenomaleas
Composition of the fatty-acid fraction of the substance :
myristic acid : maximum 5.0 per cent ;
palmitic acid : maximum 16.0 per cent ;
palmitoleic acid : maximum 8.0 per cent ;
stearic acid : maximum 6.0 per cent ;
oleic acid : minimum 58.0 per cent ;
linoleic acid : maximum 18.0 per cent ;
C24H29N3O6
Mr 455.5
linolenic acid : maximum 4.0 per cent.
DEFINITION
Ethylene oxide and dioxan (2.4.25, Method A) : maximum
(8)-9,10-Didehydro-N-[(1S)-1-(hydroxymethyl)propyl]-61 ppm of ethylene oxide and maximum 10 ppm of dioxan.
methylergoline-8-carboxamide maleate.
Water (2.5.12) : maximum 0.5 per cent, determined on 0.50 g. Content : 99.0 per cent to 101.0 per cent (dried substance).
270
CHARACTERS
Appearance : white or almost white, hygroscopic, crystalline
powder.
Solubility : soluble in water, slightly soluble in anhydrous
ethanol.
IDENTIFICATION
A. Specific optical rotation (see Tests).
Preparation : mulls.
Comparison : methylergometrine hydrogen
maleate CRS.
TESTS
pH (2.2.3) : 4.4 to 5.2.
Dilute 2.0 ml of solution S to 50.0 ml with carbon
dioxide-free water R.
Specific optical rotation (2.2.7) : + 44.0 to + 50.0 (dried
substance), determined on solution S.
Related substances. Liquid chromatography (2.2.29) : use
the normalisation procedure. Carry out the test protected
from light.
examined in 15 ml of mobile phase B and dilute to 50.0 ml
with water R.
Reference solution (a). Dissolve 1 mg of methylergometrine
hydrogen maleate impurity D CRS in 20 ml of the test
solution.
Reference solution (b). Dilute 1.0 ml of the test solution
to 100.0 ml with water R. Dilute 1.0 ml of this solution to
Reference solution (c). Dissolve 5 mg of methylergometrine

hydrogen maleate for peak identification CRS (containing
impurities A, B, C, D, E, F, G, H and I) in 3 ml of mobile
phase B and dilute to 10.0 ml with water R.
Column :
size : l = 0.10 m, = 4.6 mm ;
Mobile phase :
mobile phase A : 2 g/l solution of ammonium
carbamate R ;
mobile phase B : acetonitrile R, water R (50:50 V/V) ;
Time
(min)
0-5
Mobile phase A
(per cent V/V)
85 65
Mobile phase B
(per cent V/V)
15 35
5 - 10
65
35
10 - 15
65 20
35 80
15 - 20
20
80
20 - 21
20 85
80 15
21 - 25
85
15

Injection : 20 l.
Identification of impurities : use the chromatogram supplied
with methylergometrine hydrogen maleate for peak
identification CRS and the chromatogram obtained with
reference solution (c) to identify the peaks due to impurities
A, B, C, D, E, F, G, H and I.
Relative retention with reference to methylergometrine
(retention time = 11-12 min) : impurity A = about 0.23 ;
impurity B = about 0.46 ; impurity C = about 0.63 ;
Figure 1788.-1. Chromatogram for the test for related substances of methylergometrine hydrogen maleate spiked
with impurities A, B, C, D, E, F, G, H and I
(15) Ultracarb 5 ODS (30) or Symmetry C18 are suitable.
271
Nifuroxazide
impurity D = about 0.74 ; impurity I = about 1.10 ;

impurity E = about 1.14 ; impurity F = about 1.24 ;
impurity G = about 1.3 ; impurity H = about 1.35.
impurity D and methylergometrine.
Limits :
impurities A, B, C, D, E, G : for each impurity, maximum
0.5 per cent ;
sum of impurities F, H and I : maximum 1.0 per cent ;
sum of impurities A, B, C, D, E, F, G, H and I : maximum
1.5 per cent ;
unspecified impurities : for each impurity, maximum
0.10 per cent ;
sum of impurities other than A, B, C, D, E, F, G, H and I :
maximum 0.5 per cent ;
total: maximum 2.0 per cent ;
disregard limit : the area of the peak in the chromatogram
obtained with reference solution (b) (0.05 per cent).
on 1.0 g.
F. R = H : ergometrinine,
H. R = CH3 : methylergometrinine.
Reagents
Ammonium carbamate. CH6N2O2. (Mr 78.07). XXXXXXX.
[1111-78-0]. Carbamic acid ammonium salt.

A revision of the test for related substances is proposed to
control an additional impurity present in the batches but
not separated from the principal peak with the current
method.
XXXX:1999
NIFUROXAZIDE
ASSAY
Dissolve 0.300 g in 60 ml of anhydrous acetic acid R. Titrate
of C24H29N3O6.
STORAGE
IMPURITIES
Specified impurities : A, B, C, D, E, F, G, H, I.
A. R = H, R = CO2H : 6-methylergolene-8-carboxylic acid,

B. R = CO2H, R = H : 6-methylisoergolene-8-carboxylic acid,
C. R = H, R = CONH2 : 6-methylergolene-8-carboxamide,
E. R = CONH2, R = H : 6-methylisoergolene-8-carboxamide,
D. R1 = R2 = R3 = H, R4 = OH : ergometrine,
G. R1 = R3 = CH3, R2 = H, R4 = OH : methylsergide,
I. R1 = H, R2 = R4 = CH3, R3 = OH : ()-methylergometrine,
272
Nifuroxazidum
C12H9N3O5
Mr 275.2
DEFINITION
4-Hydroxy-N-[(1E)-(5-nitro-2-furyl)methylene]benzohydrazide.
CHARACTERS
Appearance : bright yellow, crystalline powder.
Solubility : practically insoluble in water, slightly soluble
in ethanol (96 per cent), practically insoluble in methylene
chloride.
IDENTIFICATION
Comparison : Ph. Eur. reference spectrum of nifuroxazide.
TESTS
Specific absorbance (2.2.25) : 940 to 1000 at the absorption
maximum at 367 nm.
Protected from light, dissolve 10.0 mg in 10 ml of ethylene
glycol monomethyl ether R and dilute to 100.0 ml with
methanol R. Dilute 5.0 ml of this solution to 100.0 ml with
methanol R.
Impurity A : maximum 0.05 per cent.
examined in dimethyl sulphoxide R and dilute to 10.0 ml
Test solution (b). To 5.5 ml of test solution (a) add 50.0 ml of
water R while stirring. Allow to stand for 15 min and filter.
Nifuroxazide
Reference solution. To 0.5 ml of test solution (a) add 5.0 ml

of a 50 mg/l solution of 4-hydroxybenzohydrazide R in
dimethyl sulphoxide R. Add 50.0 ml of water R while
stirring. Allow to stand for 15 min and filter.
Add 0.5 ml of phosphomolybdotungstic reagent R and
10.0 ml of sodium carbonate solution R separately to
10.0 ml of test solution (b) and to 10.0 ml of the reference
solution. Allow to stand for 1 h. Examine the 2 solutions at
750 nm. The absorbance (2.2.25) of the solution obtained
with test solution (b) is not greater than that obtained with
the reference solution.
Related substances. Liquid chromatography (2.2.29). Use
freshly prepared solutions, protected from light.
examined in 15.0 ml of dimethylformamide R and dilute
to 100.0 ml with the mobile phase. If precipitation occurs,
use the supernatant liquid.
Reference solution (a). Dissolve 10.0 mg of methyl
parahydroxybenzoate R (impurity B) in 2.0 ml of
dimethylformamide R and dilute to 20.0 ml with the mobile
phase. Dilute 1.0 ml of this solution to 100.0 ml with the
mobile phase.
Reference solution (b). Dissolve 5 mg of the substance to be
examined and 10 mg of methyl parahydroxybenzoate R in
2 ml of dimethylformamide R and dilute to 20 ml with the
mobile phase. Dilute 1 ml of this solution to 100 ml with
the mobile phase.
Column :
size : l = 0.25 m, = 4.6 mm,
stationary phase : spherical octadecylsilyl silica gel for
chromatography R (5 m) with a specific surface area
of 340 m2/g, a pore size of 10 nm and a carbon loading
of 19 per cent(16).
Mobile phase : acetonitrile R, water R (35:65 V/V).
Injection : 20 l.
Run time : 6 times the retention time of nifuroxazide.
Relative retention with reference to nifuroxazide
(retention time = about 6.5 min) : impurity A = about 0.4 ;
impurity D = about 5.2.
nifuroxazide and impurity B.
Limits :
any impurity : not more than 0.6 times the area of
the principal peak in the chromatogram obtained with
reference solution (a) (0.3 per cent), and not more than 1
such peak has an area greater than 0.2 times the area of
the principal peak in the chromatogram obtained with
reference solution (a) (0.1 per cent),
total: not more than the area of the principal peak in
(0.5 per cent),
(0.05 per cent).
Liquid chromatography (2.2.29). Use amber volumetric
flasks unless otherwise specified.
Solvent mixture : acetonitrile R, water R (40:60 V/V).

examined in the solvent mixture using sonication for not
more than 5 min and dilute to 100.0 ml with the solvent
mixture.
Reference solution (b). In order to obtain impurity E in situ,
dissolve 5 mg of the substance to be examined in the solvent
mixture in a colourless volumetric flask using sonication for
5 min and dilute to 50 ml with the solvent mixture. Leave in
ambient light for 1 h.
Reference solution (c). Dissolve 5 mg of methyl
parahydroxybenzoate CRS (impurity B) in the solvent
mixture and dilute to 100.0 ml with the solvent mixture.
Dilute 1.0 ml of this solution to 100.0 ml with the solvent
mixture.
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : spherical octadecylsilyl silica gel for
chromatography R (5 m) with a specific surface area
of 340 m2/g, a pore size of 10 nm and a carbon loading
of 19 per cent(17) ;
temperature : 10 C.
Mobile phase :
mobile phase A : tetrahydrofuran R, water R (5:95 V/V) ;
Time
(min)
0 - 10
Mobile phase A
(per cent V/V)
67
Mobile phase B
(per cent V/V)
33
10 - 30
67 43
33 57

Equilibration : 10 min with the mobile phase at the initial
composition.
Injection : 50 l.
Relative retention with reference to nifuroxazide (retention
time = about 8 min) : impurity A = about 0.36 and
0.39 ; impurity E = about 0.9 ; impurity B = about 1.2 ;
impurity C = about 2.6 ; impurity D = about 3.4.
impurity E and nifuroxazide.
Limits :
impurities B, C, D : for each impurity, not more
(0.3 per cent), and not more than 1 such peak has an area
greater than 0.2 times the area of the principal peak in
the chromatogram obtained with reference solution (c)
(0.1 per cent) ;
impurity E : not more than 0.3 times the area of the
(0.10 per cent) ;
sum of impurities other than E : not more than the area
reference solution (c) (0.5 per cent) ;
(16) Kromasil is suitable.

(17) Kromasil KR 100-5 C18 or Betasil C18 5 m are suitable.
273
Oregano

in the chromatogram obtained with reference solution (c)
(0.05 per cent).
1.0 g complies with test D. Prepare the reference solution
on 1.0 g.
ASSAY
Dissolve 0.200 g, with heating if necessary, in 30 ml of
dimethylformamide R and add 20 ml of water R. Titrate
with 0.1 M sodium hydroxide, determining the end-point
of C12H9N3O5.

The definition has been revised in order to express the
limit for thymol and carvacrol in the more usual way, with
reference to the essential oil.
In the Characters section it has been decided generally to
delete the cross reference to identification tests A and B
(macroscopic and microscopic description), since the
Identification section is mandatory while the Characters
section is only informative.
The test for foreign matter (2.8.2) is now omitted in
monographs where it is limited to 2 per cent, since this test
is covered by the general monograph for herbal drugs.
Please restrict comments to the definition only.
XXXX:1880
STORAGE
OREGANO
IMPURITIES
Specified impurities : A, B, C, D, E.
Origani herba
DEFINITION
A. R = NH-NH2 : 4-hydroxybenzohydrazide
(p-hydroxybenzohydrazide),
B. R = OCH3 : methyl 4-hydroxybenzoate,
Dried leaves and flowers separated from the stems of

Origanum onites L. or Origanum vulgare L. subsp. hirtum
(Link) Ietsw., or a mixture of both species.
Content : minimum 25 ml/kg of essential oil and minimum
1.5 per cent of carvacrol and thymol (both C10H14O ; Mr 150.2)
(anhydrous drug).
Content :
essential oil: minimum 25 ml/kg (anhydrous drug) ;
carvacrol and thymol (both C10H14O ; Mr 150.2) : minimum
60 per cent in the essential oil.
C. (5-nitro-2-furyl)methylene diacetate,
D. 5-nitro-2-furaldehyde [(1E)-(5-nitro-2-furyl)methylene]hydrazone (5-nitrofurfural azine),
E. 4-hydroxy-N-[(1Z)-(5-nitro-2-furyl)methylene]benzohydrazide (cis-nifuroxazide).
274
IDENTIFICATION
A. O. onites. The leaf is yellowish-green, usually 4-22 mm
long and 3-14 mm wide. It has a long or short petiole or
is sessile. The lamina is ovate, elliptic or ovate-lanceolate.
Margins are entire or serrate, the apex is acute or obtuse.
The veins are yellowish and conspicuous on the adaxial
surface. Flowers are solitary or seen as broken parts of
the corymb. The calyx is bract-like and inconspicuous.
The corolla is white, on top of inflorescenses or single
flowers, or inconspicuous. The bract is imbricate and
green like the leaves. The drug contains yellowish or
yellowish-brown stem parts.
O. vulgare (subsp. hirtum). The leaf is green and usually
3-28 mm long and 2.5-19 mm wide. It is petiolate or
sessile. The lamina is ovate or ovate-eliptic. The margins
are entire or serrate, the apex is acute or obtuse. Flowers
are rare, found as broken parts of the corymbs. Bracts are
greenish-yellow and imbricate. The calyx is corolla-like
and inconspicuous. The corolla is white, on top of
inflorescenses, slightly conspicuous or inconspicuous.
Oregano
B. Reduce to a powder (355). The powder is green

(O. vulgare) to yellowish-green (O. onites). Examine
under a microscope using chloral hydrate solution R.
The covering trichomes are of lamiaceous type or short,
unicellular and rarely conical ; conical trichomes shaped as
pointed teeth are more abundant in O. vulgare. Covering
trichomes are thick-walled in O. vulgare. Covering
trichomes contain prismatic crystals in O. onites, minute
needles in O. vulgare. The cuticle on covering trichomes
is smooth ; warty in O. vulgare. The epidermises of the
leaves have cells with walls that are sinuous and the
stomata are of diacytic type (2.8.3) ; in O. vulgare cells
of the upper epidermis are beaded ; secretory trichomes
with 8-16 cells (12 in O. vulgare) ; glandular trichomes are
numerous in O. onites, rare in O. vulgare. They have a
unicellular head and unicellular, bicellular or tricellular
(bicellular or tricellular in O. vulgare) stalk ; pollen grains
are smooth, spherical and more abundant in O. onites.
Top of the plate

A bluish-purple zone
_______
_______
A pale green zone
Thymol : a pink zone
A pink zone (thymol)
Carvacrol : a pale violet zone
A pale violet zone (carvacrol)
_______
_______
A pale purple zone
A grey zone
A pale green zone
A bluish-purple zone
An intense brown zone
Reference solution
Test solution
TESTS
Water (2.2.13) : maximum 120 ml/kg, determined on 20.0 g
Test solution. To 1.0 g of the powdered drug (355) add
of the powdered drug (355).
5 ml of methylene chloride R and shake for 3 min, then Total ash (2.4.16) : maximum 15.0 per cent.
filter through about 2 g of anhydrous sodium sulphate R.
Ash insoluble in hydrochloric acid (2.8.1) : maximum
4.0 per cent.
Reference solution. Dissolve 1 mg of thymol R and 10 l
of carvacrol R in 10 ml of methylene chloride R.
ASSAY
Essential oil (2.8.12). Use 30.0 g of the drug, a 1000 ml
Plate : TLC silica gel plate R.
round-bottomed flask and 400 ml of water R as the
distillation liquid. Distil at a rate of 2-3 ml/min for 2 h
without xylene R in the graduated tube.
Mobile phase : methylene chloride R.
Carvacrol and thymol. Gas chromatography (2.2.28) : use
the normalisation procedure.
Application : 20 l, as bands.
Test solution. Filter the essential oil obtained in the assay
of essential oil over a small amount of anhydrous sodium
sulphate R and dilute to 5.0 ml with hexane R by rinsing the
apparatus and the anhydrous sodium sulphate.
Drying : in air.
Reference solution. Dissolve 0.20 g of thymol R and 50 mg
of carvacrol R in hexane R and dilute to 5.0 ml with the
Detection : spray with anisaldehyde solution R using
same solvent.
10 ml for a plate 200 mm square and heat at 100-105 C
Column :
for 10 min.
material : fused silica ;
size : l = 60 m, = 0.25 mm ;
Results : see below the sequence of the zones present
stationary phase : macrogol 20 000 R (film thickness
in the chromatograms obtained with the reference
0.25 m).
solution and the test solution. Furthermore, other zones
Carrier gas : nitrogen for chromatography R or helium for
are present in the lower third and upper part of the
chromatography R.
chromatogram obtained with the test solution.
C. Thin-layer chromatography (2.2.27).
1. thymol
2. carvacrol
Figure 1880.-1 Chromatogram for the assay of carvacrol and thymol in oregano
275

Split ratio : 1:100.
Temperature :
Time
(min)
0 - 45
Column
Temperature
(C)
40 250
B. Near-infrared spectrophotometry (2.2.40).

Record the NIR spectrum of the substance to be examined
by means of a fibre-optic probe. Deviations from the
calibration spectra determined by the instrument must be
within the range specified for the model.
C. It gives reaction (a) of sodium (2.3.1).
TESTS
Solution S. Dissolve 0.10 g in water R and dilute to 10.0 ml
Detector
210
Detection : flame ionisation.
Injection: 0.2 l.
more intensely coloured than reference solution B6 (2.2.2,
Method II).
Elution order : order indicated in the composition of the
reference solution. Record the retention times of these
substances.
Optical rotation (2.2.7) : 0.4 to + 0.4.
Dissolve 0.2 g in 10 ml of water R. Adjust to pH 11.5-12.0
resolution : minimum of 1.5 between the peaks due to
with a 8 g/l solution of sodium hydroxide R. Dilute to
thymol and carvacrol.
Using the retention times determined from the chromatogram
obtained with the reference solution, locate the components
of the reference solution in the chromatogram obtained with Solvent mixture. Mix equal volumes of acetonitrile for
chromatography R and a 40 mg/l solution of sodium
the test solution.
Determine the percentage content of carvacrol and thymol. hydroxide R.
Disregard the peak of hexane.
Reference: PA/PH/Exp. P4/T (05) 11 ANP
Reference solution (b). Dissolve 23.0 mg of pantoprazole
for
peak identification CRS (containing impurities A, B, C,
Pantoprazole sodium exists in 2 stable forms :
D, E and F) in the solvent mixture and dilute to 50.0 ml with
the monohydrate and the sesquihydrate. NIR
spectrophotometry is used to distinguish between the two. the solvent mixture.
XXXX:2296 Blank solution. Solvent mixture.
Column :
PANTOPRAZOLE SODIUM
size : l = 0.125 m, = 4 mm ;
SESQUIHYDRATE
Pantoprazolum natricum sesquihydricum temperature : 40 C.
Mobile phase :
mobile phase A : 1.74 g/l solution of dipotassium
hydrogen phosphate R adjusted to pH 7.0 0.05 with a
330 g/l solution of phosphoric acid R ;
mobile phase B : acetonitrile for chromatography R ;
Injection port
190
C16H14F2N3NaO4S,1 /2H2O
1
Mr 432.4
DEFINITION
Sodium 5-(difluoromethoxy)-2-[(RS)-[(3,4-dimethoxypyridin2-yl)methyl]sulphinyl]benzimidazol-1-ide sesquihydrate.
substance).
Time
(min)
0 - 40
Mobile phase A
(per cent V/V)
80 20
Mobile phase B
(per cent V/V)
20 80
40 - 45
20 80
80 20

Detection : spectrophotometer at 290 nm and, for impurity C,
at 305 nm.
Injection : 20 l.
CHARACTERS
Appearance : white or almost white powder.
supplied with pantoprazole for peak identification CRS and
Solubility : freely soluble in water and in anhydrous ethanol, the chromatogram obtained with reference solution (b) to
identify the peaks due to impurities A, B, C, D, E and F.
practically insoluble in hexane.
Relative retention with reference to pantoprazole
IDENTIFICATION
(retention time = about 11 min) : impurity C = about 0.6 ;
impurity A = about 0.9 ; impurities D and F = about 1.2 ;
Comparison : pantoprazole sodium sesquihydrate CRS. impurity E = about 1.3 ; impurity B = about 1.5.
(18) Hypersil ODS is suitable.
276

resolution : minimum 1.5 between each consecutive peak ;
the chromatogram obtained is similar to the
chromatogram supplied with pantoprazole for peak
identification CRS.
Limits :
the peak area of impurity C by 0.3 ;
A. 5-(difluoromethoxy)-2-[[(3,4-dimethoxypyridin-2yl)methyl]sulfonyl]-1H-benzimidazole,
impurity A : not more than twice the area of the principal

sum of impurities D and F : not more than twice the area
impurities B, C, E : for each impurity, not more than the
with reference solution (a) (0.1 per cent) ;
B. 5-(difluoromethoxy)-2-[[(3,4-dimethoxypyridin-2yl)methyl]thio]-1H-benzimidazole,

total: not more than 5 times the area of the principal peak
in the chromatogram obtained with reference solution (a) C. 5-(difluoromethoxy)-1H-benzimidazole-2-thiol,
(0.5 per cent) ;
(0.05 per cent).
Sodium : 5.0 per cent to 5.8 per cent.
Atomic emission spectrometry (2.2.22, Method I).
Test solution. Dissolve 0.1 g of the substance to be examined D. 5-(difluoromethoxy)-2-[(2RS)-[(3,4-dimethoxypyridin-2yl)methyl]sulfinyl]-1-methyl-1H-benzimidazole,
in water R, add 100 l of nitric acid R and dilute to 100.0 ml
with water R.
Reference solutions. Prepare the reference solutions using
sodium standard solution (200 ppm Na) R, diluted as
necessary.
Wavelength : 330.2 nm.
0.150 g.
ASSAY
E. mixture of the 3 stereoisomers of 6,6-bis(difluoroDissolve 0.200 g in 80 ml of anhydrous acetic acid R, add
methoxy)-2,2-bis[[(3,4-dimethoxypyridin-2-yl)5 ml of acetic anhydride R and mix for at least 10 min.
methyl]sulfinyl]-1H,1H-5,5-bibenzimidazole,
Titrate with 0.1 M perchloric acid, determining the end-point
C16H14F2N3NaO4S.
STORAGE
IMPURITIES
Specified impurities : A, B, C, D, E, F.
F. 6-(difluoromethoxy)-2-[(2RS)-[(3,4-dimethoxypyridin-2yl)methyl]sulfinyl]-1-methyl-1H-benzimidazole.
277
Pholcodine
Reference: PA/PH/Exp. 11/T (02) 110 ANP 1R

This monograph has already been published in
Pharmeuropa 15.3. The LC in the test for related
substances has been modified to obtain better results :
tetrahydrofuran has been deleted from the solvent mixture
as degradation of the substance (to form N-, N- and
N,N-oxides) has been observed. Tetrahydrofuran can
be kept in the composition of the mobile phase. The
pH of the mobile phase is critical : it must be 7.90 to
prevent degradation of the column and allow separation
of impurities. It is also proposed to delete identification
tests B and C.
XXXX:0522
PHOLCODINE
Pholcodinum
C23H30N2O4,H2O
Mr 416.5
DEFINITION
7,8-Didehydro-4,5-epoxy-17-methyl-3-[2-(morpholin-4yl)ethoxy]morphinan-6-ol monohydrate.
Content : 98.5 per cent to 100.5 101.0 per cent (dried
substance).
CHARACTERS
Appearance : white or almost white, crystalline powder or
colourless crystals.
Solubility : sparingly soluble in water, freely soluble in
acetone and in ethanol (96 per cent). It dissolves in dilute
mineral acids.
IDENTIFICATION
First identification : A.
Second identification : B, C.
Comparison : Ph. Eur. reference spectrum of pholcodine
pholcodine CRS.
B. Dissolve 0.100 g in water R and dilute to 100.0 ml with
the same solvent. To 10.0 ml of this solution add 75 ml of
water R and 10 ml of 1 M sodium hydroxide and dilute
to 100.0 ml with water R. Examined between 230 nm
and 350 nm (2.2.25), the solution shows an absorption
maximum at 284 nm. The specific absorbance at the
absorption maximum is 36 to 38.
C. Dissolve 50 mg in 1 ml of sulphuric acid R and add
0.05 ml of ammonium molybdate solution R. A pale blue
colour is produced which becomes deep blue on gentle
warming. Add 0.05 ml of dilute nitric acid R. The colour
becomes brownish-red.
TESTS
substance).
Dissolve 1.000 g in ethanol (96 per cent) R and dilute to
50.0 ml with the same solvent.
(2.2.27), using silica gel G R as the coating substance.
examined in chloroform R and dilute to 10 ml with the same
solvent.
to 50 ml with chloroform R.
Reference solution (b). Dilute 5 ml of reference solution (a)
to 10 ml with chloroform R.
over a path of 15 cm using a mixture of 2.5 volumes of
concentrated ammonia R, 32.5 volumes of acetone R,
35 volumes of alcohol R and 35 volumes of toluene R. Dry
the plate in a current of air and spray with dilute potassium
iodobismuthate solution R. In the chromatogram obtained
with the test solution, any spot apart from the principal
spot is not more intense than the spot in the chromatogram
obtained with reference solution (a) (1.0 per cent) and not
more than one such spot situated above the principal spot is
more intense than the spot in the chromatogram obtained
with reference solution (b) (0.5 per cent).
0.02 M phosphate buffer solution. To 80.0 ml of 0.2 M
sodium hydroxide add 100.0 ml of 0.2 M potassium
dihydrogen phosphate R and dilute to 1000.0 ml with
water R.
Solvent mixture. Dilute 80 ml of acetonitrile R to 1000 ml
with 0.02 M phosphate buffer solution.
Reference solution (a). Dissolve 10.0 mg of codeine R in
the solvent mixture and dilute to 10.0 ml with the solvent
mixture. To 0.5 ml of this solution add 0.5 ml of the test
solution and dilute to 50.0 ml with the solvent mixture.
Column :
size : l = 0.075 m, = 4.6 mm ;
stationary phase : end-capped phenylhexylsilyl silica gel
for chromatography R (3 m)(19) with a specific surface
area of 400 m2/g and a pore size of 10 nm ;
temperature : 35 C.
Mobile phase : to 50 ml of tetrahydrofuran R add 75 ml of
acetonitrile R and dilute to 1000 ml with 0.02 M phosphate
buffer solution ; adjust to pH 7.90 with 0.2 M sodium
hydroxide ; the pH must not exceed 8.0.
Injection : 20 l.
Run time : 4 times the retention time of pholcodine.
Relative retention with reference to pholcodine
impurity B = about 0.8 ; impurity D = about 2.3.
(19) Phenomenex Luna is suitable.
278
Pholcodine
1. impurity F
2. impurity C
3. impurity A
4. impurity B
5. pholcodine
6. impurity D
Figure 0522.-1. Chromatogram for the test for related substances of pholcodine : pholcodine containing N-oxides,
morphine and codeine

impurity B and pholcodine.
Limits :
impurities A, B, D : for each impurity, not more than
twice the area of the principal peak in the chromatogram

on 1.0 g.
ASSAY
Dissolve 0.180 g in 50 ml of anhydrous acetic acid R,
warming gently. Titrate with 0.1 M perchloric acid,
determining the end-point potentiometrically (2.2.20) at the
2nd point of inflexion.
of C23H30N2O4.
IMPURITIES
total: not more than 7 times the area of the principal peak
in the chromatogram obtained with reference solution (b) Specified impurities : A, B, D.
(0.7 per cent) ;
(0.05 per cent).
Morphine. Dissolve 0.10 g in 0.1 M hydrochloric acid and
dilute to 5 ml with the same acid. Add 2 ml of a 10 g/l
solution of sodium nitrite R, allow to stand for 15 min
and add 3 ml of dilute ammonia R1. The solution is not
pharmaceutical use) : C, E, F.
more intensely coloured than reference solution B4 (2.2.2,
Method II) (about 0.13 per cent of morphine).
A. morphine,
Loss on drying (2.2.32) : 3.9 per cent to 4.5 per cent,
determined on 0.500 g by drying in an oven at 100-105 C.
B. codeine,
279
2.9.44. Preparations for nebulisation : characterisation
Reference: PA/PH/Exp. INH/T (05) 2 ANP

NOTE ON THE GENERAL CHAPTER
The European Pharmacopoeia monograph on
Preparations for inhalation (0671), under the section
on Liquid preparations for nebulisation, states that the
Continuously operating nebulisers ... allow the dose to
C. (17RS)-7,8-didehydro-4,5-epoxy-17-methyl-3-[2-(morpholin- be inhaled at an appropriate rate and particle size which
4-yl)ethoxy]morphinan-6-ol 17-oxide (pholcodine N-oxide), ensures deposition of the preparation in the lungs. The
CEN (European Committee for Standardisation) worked
on the assessment of the capabilities of the nebulisers
themselves ; but in response to the requirements of the
European Pharmacopoeia monograph, it was considered
useful to set up a methodology for the control of the
particle/droplet size and particle/droplet size distribution.
Currently, neither the European Pharmacopoeia nor the
USP contain a chapter describing a modern methodology to
D. unknown structure,
be used for the assessment of preparations for nebulisation.
The following text results from international efforts
to draft a new chapter for both pharmacopoeias. The
methods proposed below have been tested in a number of
laboratories and a collaborative study, planned amongst
EPAG (European Pharmaceutical Aerosol Group) members,
will take place in 2006.
E. 7,8-didehydro-4,5-epoxy-17-methyl-3-[2-(4-oxidomorpholin4-yl)ethoxy]morphinan-6-ol (pholcodine N-oxide),
The general chapter is divided into two parts : active

substance delivery rate and total active substance delivered,
and Aerodynamic assessment of nebulised aerosols. Based
on the requirements of the Quality Working Party-health
Canada Guideline on Pharmaceutical Quality of Inhalation
and Nasal Products, it is suggested to restrict the use of
the first method (active substance delivery rate and total
active substance delivered) to the development phase
of the preparation for nebulisation ; the second method
(Aerodynamic assessment of nebulised aerosols) would be
for use in routine control. Readers are invited to comment
on this restriction. The same text should be published in
Pharmacopoeial Forum 32(4) as a Stimuli article.
XXXX:20944
2.9.44. PREPARATIONS FOR

NEBULISATION : CHARACTERISATION
Products used for nebulisation and intended for pulmonary
delivery are characterised using the following tests :
F. (17RS)-7,8-didehydro-4,5-epoxy-17-methyl-3-[2-(4oxidomorpholin-4-yl)ethoxy]morphinan-6-ol 17-oxide
(pholcodine N,N-dioxide).
Reagents
Silica gel for chromatography, phenylhexylsilyl,
end-capped. XXXXXXX.
A very finely divided silica gel (3 m), chemically modified
at the surface by the bonding of phenylhexyl groups. The
particle size is indicated after the name of the reagent in the
tests where it is used.
Active substance delivery rate and total active substance

delivered ;
Aerodynamic assessment of nebulised aerosols.
These tests standardise the approach given to the assessment
of the dose that would be delivered to a patient but are not
intended to provide assessment of the nebuliser device itself,
which is described in a CEN standard(20).
The mass- rather than the number-weighted size distribution
is more appropriate to evaluate product performance, since
active substance mass as a function of aerodynamic diameter
is more indicative of therapeutic effect within the respiratory
tract.
(20) European Standard 13544-1:2001. Respiratory Therapy Equipment. Part 1 : Nebulising systems and their components. European Committee for Standardization. Brussels, Belgium. 2001.
280
ACTIVE SUBSTANCE DELIVERY RATE AND TOTAL

ACTIVE SUBSTANCE DELIVERED
These tests are performed to assess the total active substance
delivered to a patient and the rate of delivery to the patient
using standardised conditions of volumetric flow rate.
It is essential that breath-enhanced and breath-actuated
nebulisers be evaluated by a breathing simulator, as the
output of these types of device is highly dependent on
inhalation flow rate. The methodology below describes
the use of a standard breathing pattern defined for adults.
Should a particular product for nebulisation only be
indicated for paediatric use, then paediatric breathing
pattern(s) must be used(21). Breathing patterns are used,
rather than continuous flow rates, to provide a more
appropriate measure of the mass of active substance that
would be delivered to patients.
Active substance delivery rate and total active substance
delivered are appropriate characteristics because they
allow the mass delivered to be characterised in a standard
way regardless of the nebuliser used. Accordingly, the test
methodology described below allows that the mass of active
substance delivered in the first period (typically 1 min) is
measured (consequently giving an assessment of active
substance delivery rate) as well as capturing the total active
substance mass delivered.
APPARATUS
Breath simulator. A commercially available breath
simulator(22), which is able to generate the breathing profile
specified in Table 2.9.44.-1, is used for the test. The pattern is
intended to simulate an adult breathing pattern. A different
breathing pattern may be used where appropriate, e.g.
medicinal products specifically intended for use in children
may be tested with a breathing pattern more appropriate
for a child.
Table 2.9.44.-1. Breath simulator specification
Item
Specification
Tidal volume
500 ml
Frequency
15 cycles per min
Waveform
sinusoidal
inhalation:exhalation ratio
1:1
Filter system. A suitably validated low-resistance filter(23),

capable of quantitatively collecting the aerosol and enabling
recovery of the active substance with an appropriate solvent,
is used for the test. The dead volume of the filter casing
does not exceed 10 per cent of the tidal volume used in the
breath simulation.
A. inhalation filter and filter holder
B. breath simulator
C. nebuliser
Figure 2.9.44.-1. Experimental set-up for breath simulator

testing
Start the breath simulator then, at the beginning of an
inhalation cycle, start the nebuliser. Operate the nebuliser
for a defined initial time period. The length of the time
interval ensures that sufficient active substance is deposited
on the inhalation filter for quantitative analysis. A time of
60 1 s typically enables direct determination of the active
substance delivery rate. If necessary, the length of the time
interval for aerosol collection can be increased if there is
insufficient active substance deposition on the inhalation
filter to allow quantitative analysis over the 60 s period. At
the end of this initial period, stop the nebuliser, dismantle
the filter holder and recover the active substance from the
filter and the filter holder using a suitable solvent.
Place a fresh filter and filter holder in position and continue
until nebulisation ceases. Interrupt nebulisation and
exchange filters if necessary, to avoid filter saturation. The
end of nebulisation for gas-powered nebulisers is 1 min after
the beginning of sputtering, and for electronic nebulisers at
the end of operation.
Table 2.9.44.-2. Environmental conditions
Item
Specification range
Temperature
23 2 C
Pressure
86-106 kPa
Relative humidity
45-75 per cent
RESULTS
Using a suitable method of analysis, determine the mass of
active substance collected on the filters during each time
interval. Determine the active substance delivery rate by
dividing the mass of active substance collected on the first
inhalation filter by the time interval used for collection.
Determine the total mass of active substance delivered
by summing the mass of active substance collected on all
inhalation filters.
AERODYNAMIC ASSESSMENT OF NEBULISED

AEROSOLS
Nebulised products need to be size-characterised at flow
rates lower than the range that is normally used for
powder inhalers and metered-dose inhalers. A flow rate
of 15 litres/min is recommended in the CEN standard
METHOD
because this value represents a good approximation to the
Attach the filter (contained in the filter holder) (A) to the
mid-inhalation flow rate achievable by a tidally-breathing
breath simulator (B) according to Figure 2.9.44.-1. Fill
healthy adult (500 ml tidal volume).
the nebuliser with the volume of the medicinal product as
specified in the patient instructions. Attach the mouthpiece Although low-angle laser light scattering instruments
(laser diffractometers) can provide rapid size-distribution
of the nebuliser to the inhalation filter using a mouthpiece
measurements of nebuliser-generated aerosols, these
adapter if required, ensuring that connections are airtight.
Make sure the nebuliser is positioned in the same orientation techniques do not detect the active substance, rather they
measure the size distribution of the droplets irrespective of
as intended for use. This may require tilting the breath
their content. This may not be a problem with homogeneous
simulator and filter holder. Ensure that environmental
solutions, but can result in significant error if the product
conditions are within the limits specified in Table 2.9.44.-2.
Set the breath simulator to generate the specified breathing to be nebulised is a suspension, or if droplet evaporation is
significant as can be the case with certain nebuliser types.
pattern.
(21) Suitable breathing patterns for paediatric use may be found for example in Canadian Standard CAN/CSA/Z264.1-02:2002, Spacers and Holding Chambers for Use with Metered Dose
Inhalers. Canadian Standards Association, Mississauga, Canada 2002.
(22) A suitable model is the Pari Compass, (PARI, Steinerstr. 15k, D-81369 Munich, Germany).
(23) e.g. product K 248 from 3M or 041B0523 from Pari, or equivalent.
281
Re-entrainment. Although droplet bounce and

Cascade impactors enable the aerosol to be characterised
unambiguously in terms of the mass of active substance as a re-entrainment is less likely with nebuliser-produced droplets
than with solid particles from other types of inhaler, it is
function of aerodynamic diameter.
important that assessing the need for coating be part of
Apparatus E, a cascade impactor, has been calibrated at
method development. It may be omitted where justified
15 litres/min specifically to meet the recommendation of the and authorised. Where necessary to ensure efficient
CEN Standard, and is therefore used for this test(24). It is
particle capture, coat each cup with silicone oil or a similar
also recognised that the control of evaporation of droplets
high-viscosity liquid, typically deposited from a volatile
produced by nebulisers may be critical to avoid bias in the
solvent.
droplet size assessment process. The method describes an
Impactor configuration. A back-up filter in addition to
approach in which evaporation is standardised by operating
the micro-orifice collector (MOC) must be used to ensure
with air or another driving gas that is within a defined range
quantitative recovery of active substance from the nebulised
of water vapour content. Determining mass balance in the
aerosol at the specified flow rate of 15 litres/min. The
same way as for powder inhalers and metered-dose inhalers
MOC may be removed for the purpose of nebuliser testing.
is not straightforward, in that the dose is being captured as
However, if is retained, either the filter is located below the
a continuous output, and hence is not included. As part
MOC (internal filter option) or a filter used, that is external
of method development, recovery experiments must be
to the impactor is used, to capture any fine droplets that pass
performed to validate the method.
beyond the last size fractionating stage. The pre-separator is
not used for testing nebuliser-generated aerosols.
The droplets produced by certain types of nebuliser (i.e.
non-air entrainment), may be more susceptible to evaporation METHOD
caused by heat transfer from the impactor during the
Locate the test system in an environment in which the
measurement process(25). As part of method development,
ambient air is at a temperature of 23 2 C at a relative
it is good practice to check if the nebulised aerosol is
humidity of 45-75 per cent (or cool the impactor to 5-10 C,
significantly affected by evaporation, by comparing the size if method development has demonstrated the need to control
distribution of the aerosol using an impactor operated at
droplet size changes caused by heat transfer to the incoming
ambient temperature compared with a cooled impactor. If
aerosol by the impactor).
there is a significant difference, the operating conditions
Attach the induction port to the impactor, and connect the
described below in the chapter may not be suitable. In these outlet of the impactor to a vacuum source that is capable
cases, cool the impactor to 5-10 C before proceeding with
of withdrawing air through the system at 15 litres/min as
the measurements of the droplet size distribution.
specified in Figure 2.9.44.-2. Locate a 3-way gate valve
between the impactor and the vacuum source, and set the
APPARATUS
A detailed description of Apparatus E and the induction port valve so that the flow is sampled from the impactor. Insert
is contained in chapter 2.9.18, and includes details of critical a suitable filter (e.g. a GF/A glass fibre filter), capable of
quantitatively collecting the active substance in either the
dimensions and the qualification process for the impactor
internal or external filter holder.
(stage mensuration).
Connect a flow meter, calibrated for the volumetric flow
METHOD VALIDATION
leaving the meter, to the induction port. Adjust the flow
control valve located between the impactor and three-way
Impactor stage overloading. During method development
valve to achieve a steady flow through the system at
and validation, it is important to confirm that the volume
15 litres/min ( 5 per cent). Turn off the flow and set the
of liquid sampled from the nebuliser does not overload the
3-way valve so that the flow to the vacuum source bypasses
impactor. Visual inspection of the collection surfaces on
the impactor. Remove the flow meter.
stages collecting most of the droplets will normally reveal
streaking if overloading has occurred. This phenomenon is Set up the nebuliser with a supply of driving gas (usually
air or oxygen) at the pressure and flow rate specified by
usually also associated with an increase in mass of active
the manufacturer. Take precautions to ensure that the gas
substance collected on the final stage and back-up filter.
supply line does not become detached from the nebuliser
Reducing the sampling period (T0) is the most effective
when under pressure. Fill the nebuliser with the volume of
way to avoid overloading in any given system, balancing
the medicinal product as specified in the patient instructions.
overloading with analytical sensitivity.
A. nebuliser
C. apparatus E
E. three-way valve
B. induction port
D. control valve
F. vacuum source
Figure 2.9.44.-2. Apparatus E for measuring the size distribution of preparations for nebulisation
(24) Marple VA, Olson BA, Santhanakrishnan K et al. Next generation pharmaceutical impactor : A new impactor for pharmaceutical inhaler testing. Part III : Extension of archival calibration to
15 l/min. J Aerosol Med 2004; 17(4):335-343.
(25) Finlay WH, Stapleton KW. Undersizing of droplets from a vented nebuliser caused by aerosol heating during transit through an andersen impactor. J Aerosol Sci 1999 ; 30(1):105-109.
282
Set the 3-way valve so that the flow bypasses the impactor.
Switch on the flow from the vacuum source downstream
of the impactor then switch on the flow of driving gas to
the nebuliser at the steady pre-determined value. When the
flow rate is stable, attach the mouthpiece of the nebuliser to
the induction port, using a mouthpiece adapter if required,
ensuring that the mouthpiece is aligned on-axis with the
induction port. Set the 3-way valve so that the flow passes
through the impactor. Sample for a pre-determined time (T0).
Once determined, this time (T0) must be defined and used
in the analytical method for a particular medicinal product
to ensure that mass fraction data can be compared. At the
end of the sampling period, remove the nebuliser from the
induction port and switch off the flow from the vacuum
source to the impactor. Switch off the driving gas flow to
the nebuliser.
Dismantle the impactor and, using a suitable method of
analysis, determine the mass of active substance collected in
the induction port, on each stage and on the back-up filter as
described for Apparatus E (2.9.18). If the MOC is retained,
add the mass of active substance collected in the MOC to
that deposited on the back-up filter and treat as a single
sample for the purpose of subsequent calculations.
Calculate the mass fraction (Fm,comp) of the active substance
deposited on each component of the impactor, commencing
with the induction port and proceeding in order through the
impactor, using the following expression:
mcomp =
M
for the appropriate cut-off diameters at 15 litres/min).

Plot the cumulative fraction of active substance versus
cut-off diameter in a suitable format, e.g. logarithmic
or log-probability format (see Figure 2.9.44.-4). Where
appropriate, use this plot to determine by interpolation the
fraction either less than a given size or between an upper
and lower size limit.
Table 2.9.44.-3. Cut-off sizes for Apparatus E at
15 litres/min
Stage
Cut-off diameter (m)
14.1
8.61
5.39
3.30
2.08
1.36
0.98
mass associated with the component under

evaluation ;
total mass collected by the system.
Present Fm,comp in order of location within the measurement

equipment, beginning at the induction port and ending with
the back-up filter of the impactor (see Figure 2.9.44.-3).
Where appropriate, Fm,comp for adjacent stages of the impactor
may be combined in order to report the mass fraction
collected on a group of stages as a single value.
Figure 2.9.44.-4. Cumulative mass-weighted size

distribution from apparatus E-measured data :
GSD = (d84.1/d15.9) for a log-normal size distribution
If necessary, and where appropriate, use this plot to
determine values for the MMAD and the GSD, as appropriate.
Appropriate computational methods may also be used.

The TLC test for related substances has been replaced by
an LC test. Only comments on this test are requested.
XXXX:0524
PROMETHAZINE HYDROCHLORIDE
Promethazini hydrochloridum
Figure 2.9.44.-3. Example of mass fraction of droplets
presented in terms of location within the sampling system
Determine the cumulative mass-weighted particle-size
distribution of the aerosol size-fractionated by the impactor
in accordance with the procedure given in chapter 2.9.18.
Starting at the filter, derive a cumulative mass versus
cut-off diameter of the respective stages (see Table 2.9.44.-3
C17H21ClN2S
Mr 320.9
283
DEFINITION
(2RS)-N,N-Dimethyl-1-(10H-phenothiazin-10-yl)propan-2amine hydrochloride.
CHARACTERS
Appearance : white or faintly yellowish, crystalline powder.
Solubility : very soluble in water, freely soluble in ethanol
(96 per cent) and in methylene chloride.
mp : about 222 C, with decomposition.
solution (b) (0.5 per cent) and at most three such spots are
with reference solution (c) (0.2 per cent).
Liquid chromatography (2.2.29). Carry out the test protected
from light and use freshly prepared reference solutions (a)
and (b).
The following chromatogram is shown for information but
will not be published in the European Pharmacopoeia.
IDENTIFICATION
First identification : A, B, D.
Comparison : promethazine hydrochloride CRS.
B. It complies with the identification test for phenothiazines
by thin-layer chromatography (2.3.3).
C. Dissolve 0.1 g in 3 ml of water R. Add dropwise 1 ml
of nitric acid R. A precipitate is formed which rapidly
dissolves to give a red solution, becoming orange and
then yellow. Heat to boiling. The solution becomes
orange and an orange-red precipitate is formed.
D. It gives reaction (b) of chlorides (2.3.1).
TESTS
pH (2.2.3) : 4.0 to 5.0, measured immediately after
preparation.
Dissolve 1.0 g in carbon dioxide-free water R and dilute to
10 ml with the same solvent.
Related substances. Carry out the test protected from
bright light. Prepare the solutions immediately before use.
Examine by thin-layer chromatography (2.2.27), using a
suitable silica gel as the coating substance.
examined in a mixture of 5 volumes of diethylamine R and
95 volumes of methanol R and dilute to 10 ml with the same
mixture of solvents.
Reference solution (a). Dissolve 20 mg of isopromethazine
hydrochloride CRS in a mixture of 5 volumes of
diethylamine R and 95 volumes of methanol R and dilute to
100 ml with the same mixture of solvents.
100 ml with a mixture of 5 volumes of diethylamine R and
95 volumes of methanol R.
Reference solution (c). Dilute 0.2 ml of the test solution to
100 ml with a mixture of 5 volumes of diethylamine R and
95 volumes of methanol R.
Apply separately to the plate 10 l of the test solution and
10 l of each reference solution. Develop in an unsaturated
tank over a path of 12 cm using a mixture of 5 volumes of
diethylamine R, 10 volumes of acetone R and 85 volumes
of cyclohexane R. Allow the plate to dry in air and examine
in ultraviolet light at 254 nm. Disregard any spot at the
starting point.
In the chromatogram obtained with the test solution : any
spot corresponding to isopromethazine hydrochloride is not
more intense that the spot in the chromatogram obtained
with reference solution (a) (1 per cent) ; any spot, apart
from the principal spot and the spot corresponding to
isopromethazine hydrochloride, is not more intense than
the spot in the chromatogram obtained with reference
1. impurity D
3. promethazine
2. impurity C
4. impurity B
5. impurity A
Figure 0524.-1. Chromatogram for the test for

related substances of promethazine hydrochloride :
solution of promethazine hydrochloride spiked
with impurities A, B, C and D
Solvent mixture : triethylamine R, methanol R (1:1000 V/V).
Reference solution (a). Dissolve 5.0 mg of promethazine
hydrochloride for peak identification CRS (containing
impurities A, B and C) in the solvent mixture and dilute
Reference solution (c). In order to prepare in situ the
degradation compound (impurity D), to 1.5 ml of the test
solution add 50 l of strong hydrogen peroxide solution R.
Allow to stand for 5 h at room temperature.
Column :
size : l = 0.15 m, = 4.6 mm ;
stationary phase : end-capped octylsilyl silica gel for
chromatography with polar incorporated groups R
(5 m)(26).
Mobile phase : mix 20 volumes of methanol R, 30 volumes
of acetonitrile R and 50 volumes of a 3.4 g/l solution of
potassium dihydrogen phosphate R previously adjusted to
pH 7.0 with potassium hydroxide R.
Injection : 10 l.
(26) Symmetry shield RP8 is suitable.
284
Pyridoxine hydrochloride
Run time : twice the retention time of promethazine.

Relative retention with reference to promethazine
(retention time = about 18 min) : impurity D = about 0.2 ;
impurity C = about 0.5 ; impurity B = about 1.4 ;
IMPURITIES
System suitability :
impurities B and A in the chromatogram obtained with
reference solution (a) ;
identification of impurities : use the chromatogram
supplied with promethazine hydrochloride for peak
identification CRS and the chromatogram obtained
with reference solution (a) to identify the peaks due to
impurities A, B and C ; use the chromatogram obtained
with reference solution (c) to identify the peak due to
impurity D.
A. phenothiazine,
Limits :
impurity D = 5 ;
B. (2RS)-N,N-dimethyl-2-(10H-phenothiazin-10-yl)propan-1amine (isopromethazine),
impurities A, D : for each impurity, not more than 0.1 times

impurities B, C : for each impurity, not more than 0.2 times
(0.10 per cent) ;
total: not more than 0.6 times the area of the principal
solution (b) (0.6 per cent) ;
C. R = H, X = S : (2RS)-N-methyl-1-(10H-phenothiazin-10yl)propan-2-amine,
D. R = CH3, X = SO : (2RS)-N,N-dimethyl-1-(10H-phenothiazin10-yl)propan-2-amine S-oxide.

(0.05 per cent).
Reference: PA/PH/Exp. VIT/T (05) 14 ANP
Dissolve 1.0 g in 5 ml of water R, then add 5 ml of acetone R

and 5 ml of buffer solution pH 3.5 R. Carry out the
It is proposed to replace the TLC test for related substances
prefiltration. The prefiltrate complies with test E. Prepare
the reference solution using 5 ml of lead standard solution with an LC test. Only comments on this test are requested.
XXXX:0245
(2 ppm Pb) R.
PYRIDOXINE HYDROCHLORIDE

on 1.0 g.
Pyridoxini hydrochloridum
ASSAY
Dissolve 0.250 g in a mixture of 5.0 ml of 0.01 M hydrochloric
acid and 50 ml of ethanol (96 per cent) R. Carry out
a potentiometric titration (2.2.20), using 0.1 M sodium
hydroxide. Read the volume added between the 2 points
of inflexion.
of C17H21ClN2S.
STORAGE
C8H12ClNO3
Mr 205.6
DEFINITION
(5-Hydroxy-6-methylpyridine-3,4-diyl)dimethanol
hydrochloride.
285
Pyridoxine hydrochloride
CHARACTERS
Solubility : freely soluble in water, slightly soluble in ethanol
(96 per cent).
mp : about 205 C, with decomposition.
IDENTIFICATION
First identification : B, D.
Second identification : A, C, D.
A. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Solution A. Dilute 1.0 ml of solution S (see Tests) to
50.0 ml with 0.1 M hydrochloric acid.
Solution B. Dilute 1.0 ml of solution A to 100.0 ml with
0.1 M hydrochloric acid.
Solution C. Dilute 1.0 ml of solution A to 100.0 ml with
the potassium dihydrogen phosphate 0.025 M + disodium
hydrogen phosphate 0.025 M solution described in
chapter 2.2.3.
Spectral ranges : 250-350 nm for solution B, 220-350 nm
for solution C.
Absorption maxima: 288-296 nm for solution B,
248-256 nm and 320-327 nm for solution C.
Specific absorbances at the absorption maxima :
at 288 nm to 296 nm : 425 to 445 for solution B ;
at 248 nm to 256 nm : 175 to 195 for solution C ;
at 320 nm to 327 nm : 345 to 365 for solution C.
Comparison : pyridoxine hydrochloride CRS.
C. Thin-layer chromatography (2.2.27).
examined in water R and dilute to 10 ml with the same
solvent.
with water R.
Reference solution (a). Dissolve 0.10 g of pyridoxine
hydrochloride CRS in water R and dilute to 10 ml with
the same solvent.
Reference solution (b). Dilute 2.5 ml of test solution (a)

to 100 ml with water R. Dilute 1 ml of this solution to
10 ml with water R.
Plate : TLC silica gel G plate R.
Mobile phase : concentrated ammonia R,
methylene chloride R, tetrahydrofuran R, acetone R
(9:13:13:65 V/V/V/V).
Application : 2 l.
Development : in an unsaturated tank, over a path of
15 cm.
Drying : in air.
Detection : spray with a 50 g/l solution of sodium
carbonate R in a mixture of 30 volumes of ethanol
(96 per cent) R and 70 volumes of water R ; dry
in a current of air, spray with a 1 g/l solution of
dichloroquinonechlorimide R in ethanol (96 per cent) R
and examine immediately.
with test solution (b) is similar in position, colour and size
to the principal spot in the chromatogram obtained with
reference solution (a).
TESTS
more intensely coloured than reference solution Y7 (2.2.2,
Method II).
(2.2.27), using a TLC silica gel G plate R.
solvent.
with water R.
Reference solution (a). Dissolve 0.10 g of pyridoxine
hydrochloride CRS in water R and dilute to 10 ml with the
same solvent.
1. pyridoxine
2. impurity A
3. impurity B
Figure 0245.-1. Chromatogram for the test for related substances of pyridoxine hydrochloride : solution of pyridoxine
hydrochloride spiked with impurities A and B
286
Reference solution (b). Dilute 2.5 ml of test solution (a) to

100 ml with water R. Dilute 1 ml of this solution to 10 ml
with water R.
Apply to the plate 2 l of each solution. Develop in an
unsaturated tank over a path of 15 cm using a mixture of
9 volumes of concentrated ammonia R, 13 volumes of
methylene chloride R, 13 volumes of tetrahydrofuran R
and 65 volumes of acetone R. Allow the plate to dry in air
and spray with a 50 g/l solution of sodium carbonate R in
a mixture of 30 volumes of alcohol R and 70 volumes of
water R. Dry the plate in a current of air, spray with a 1 g/l
solution of dichloroquinonechlorimide R in alcohol R and
examine the chromatograms immediately. Any spot in the
chromatogram obtained with test solution (a), apart from
the principal spot, is not more intense than the spot in the
chromatogram obtained with reference solution (b) (0.25 per
cent). Disregard any spots remaining on the starting line.
solvent.
to 100.0 ml with water R. Dilute 1.0 ml of this solution to
Reference solution (b). Dissolve 5.0 mg of pyridoxine
hydrochloride for system suitability CRS (containing
impurities A and B) in water R and dilute to 2 ml with the
same solvent.
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : base-deactivated end-capped
octadecylsilyl silica gel for chromatography R (5 m)(27).
Mobile phase : mix equal volumes of a 1.7905 g/l solution of
disodium hydrogen phosphate R and a 0.69 g/l solution of
sodium dihydrogen phosphate monohydrate R.
Injection : 5 l.
Run time : 1.5 times the retention time of pyridoxine.
with pyridoxine hydrochloride for system suitability CRS
to identify the peaks due to impurities A and B.
Relative retention with reference to pyridoxine
pyridoxine and impurity A and minimum 1.5 between the
peaks due to impurities A and B.
Limits :
impurities A, B : for each impurity, not more than the
with reference solution (a) (0.1 per cent) ;
total: maximum 0.2 per cent ;
(0.05 per cent).

12 ml of solution S complies with test A. Prepare the reference
solution using lead standard solution (1 ppm Pb) R.
on 1.0 g.
ASSAY
In order to avoid overheating in the reaction medium, mix
thoroughly throughout and stop the titration immediately
after the end-point has been reached.
Dissolve 0.150 g in 5 ml of anhydrous formic acid R. Add
50 ml of acetic anhydride R. Titrate with 0.1 M perchloric
acid, determining the end-point potentiometrically (2.2.20).
Carry out a blank titration.
C8H12ClNO3.
STORAGE
IMPURITIES
Specified impurities : A, B.
A. 6-methyl-1,3-dihydrofuro[3,4-c]pyridin-7-ol,
B. 5-(hydroxymethyl)-2,4-dimethylpyridin-3-ol.

XXXX:2325
RABBIT HAEMORRHAGIC DISEASE

VACCINE (INACTIVATED)
Vaccinum morbi haemorrhagici cuniculi
inactivatum
1. DEFINITION
Rabbit haemorrhagic disease vaccine (inactivated) is a
preparation of a suitable strain of rabbit haemorrhagic
disease virus (RHDV). This monograph applies to
vaccines intended for administration to rabbits for active
immunisation.
2. PRODUCTION
2-1. PREPARATION OF THE VACCINE
The vaccine virus is grown in rabbits. The rabbits must be
healthy, from a breeding unit, not vaccinated and not treated
with antibiotics, and free from antibodies against RHDV.
A suspension is prepared from a homogenate of suitable
internal organs of those rabbits that succumb to the infection
(27) Hypersil BDS C18 is suitable.
287
within 96 h of inoculation. The virus in the suspension may

be purified and concentrated, and is inactivated by a suitable
method.
2-2. SEED LOTS
2-2-1. Extraneous agents. Each master seed lot complies
with the tests for extraneous agents in seed lots prescribed
in the monograph Vaccines for veterinary use (0062).
2-3. CHOICE OF VACCINE COMPOSITION
The vaccine is shown to be satisfactory with respect to
safety (5.2.6) and efficacy (5.2.7) for the rabbits for which
it is intended. The following tests may be used during the
demonstration of safety (2-3-1.) and immunogenicity (2-3-2.).
2-3-1. Safety. The test is carried out for each route of
administration to be stated on the label. Use not less than
10 healthy rabbits from a breeding unit, not older than
the minimum age to be recommended for vaccination and
free from antibodies against RHDV. Administer to each
rabbit by a recommended route and method 2 doses of the
vaccine. Observe the animals for 21 days. Record the body
temperature the day before vaccination, at vaccination,
4 h after vaccination and then daily for 4 days ; note
the maximum temperature increase for each animal. No
abnormal local or systemic reaction occurs ; the average
body temperature increase does not exceed 1.5 C and no
animal shows a temperature rise greater than 2 C. If the
vaccine is intended for use in pregnant rabbits, administer
the vaccine to not less than 10 pregnant rabbits according to
the schedule to be recommended on the label. Prolong the
observation period until 1 day after parturition. The rabbits
remain in good health and there is no abnormal local or
systemic reaction. No adverse effects on the pregnancy or
on the offspring are noted.
2-3-2. Immunogenicity. The test is carried out for each route
of administration to be stated on the label. Use not less than
15 susceptible healthy rabbits not less than 10 weeks old,
from a breeding unit, free from antibodies against RHDV and
reared in suitable isolation conditions to ensure absence of
contact with RHDV. Administer 1 dose of vaccine to each of
not less than 10 of the rabbits according to the instruction
for use to be stated on the label. Keep not less than 5 other
rabbits as controls. Not less than 21 days after vaccination,
administer by a suitable route to each rabbit a quantity of a
virulent strain of RHDV sufficient to cause typical signs of
rabbit haemorrhagic disease (RHD) in a susceptible rabbit.
Observe the rabbits for a further 14 days.
The test is not valid if, after challenge, less than 80 per cent
of control rabbits die with typical signs of RHD within 96 h
of challenge.
The vaccine complies with the test if not less than 90 per
cent of vaccinated rabbits show no signs of RHD.
2-4. MANUFACTURERS TESTS
2-4-1. Residual live virus. A test for residual live virus is
carried out on the bulk harvest of each batch to confirm
inactivation of the RHDV. The test for inactivation is carried
288
out in healthy, susceptible rabbits, not less than 10 weeks

old, from a breeding unit and free from antibodies against
RHDV. 5 rabbits are inoculated by a suitable parenteral route
(subcutaneous or intramuscular) with at least a 5 ml dose
of the suspension, and 5 unvaccinated rabbits are kept as
controls. The rabbits are observed for not less than 7 days. At
the end of the observation period, the animals are euthanised
and liver and spleen extracts are tested by a suitable method
for freedom from RHDV. The vaccine complies with the test if
no rabbit dies or shows clinical signs of RHD and no RHDV
antigen is detected in the livers and spleens.
2-4-2. Batch potency test. It is not necessary to carry
out the Potency test (section 3-4.) for each batch of the
vaccine if it has been carried out using a batch of vaccine
with a minimum potency. Where the test is not carried out,
an alternative validated method is used, the criteria for
acceptance being set with reference to a batch of vaccine
that has given satisfactory results in the test described under
Immunogenicity (section 2-3-2.).
The following method is given as an example. Administer
1 dose of vaccine intramuscularly to each of 5 healthy
rabbits, 10 weeks old, from a breeding unit and free from
antibodies against RHDV. Keep 2 rabbits as unvaccinated
controls. Collect serum samples from each rabbit just before
administration of the vaccine and after the period defined
when testing the reference vaccine ; determine the antibody
titre of each serum by a suitable serological method, for
example, ELISA. The antibody levels are not significantly less
than those obtained with a batch that has given satisfactory
results in the test described under Potency (reference
vaccine). The test is not valid unless the sera collected from
the unvaccinated controls and from the rabbits just before
the administration of the vaccine are free from detectable
specific antibody.
3. BATCH TESTS
3-1. Identification. In susceptible animals, the vaccine
stimulates the production of specific antibodies against
RHDV, detectable by a haemagglutination-inhibition test or
enzyme immunoassay.
3-2. Bacteria and fungi. The vaccine complies with the
test for sterility prescribed in the monograph Vaccines for
veterinary use (0062).
3-3. Safety. Use 2 healthy rabbits, not less than 10 weeks
old, from a breeding unit and free from antibodies against
RHDV. Administer by a recommended route to each rabbit
2 doses of vaccine. Observe the rabbits for 21 days. The
vaccine complies with the test if no rabbit shows notable
clinical signs of disease or dies from causes attributable to
the vaccine.
3-4. Potency. The vaccine complies with the requirements of
the test mentioned under Immunogenicity (section 2-3-2.),
when administered by a recommended route and method. It
is not necessary to carry out the Potency test for each batch
of vaccine if it has been carried out on a representative batch.
Riboflavin
Reference: PA/PH/Exp. VIT/T (05) 16 ANP
1st application : 2 l of methylene chloride R then 2 l

of the test solution ;
2nd application : 2 l of methylene chloride R then
The TLC test for related substances has been replaced by
2 l of reference solution (a) ;
an LC test.
3rd application : 2 l of reference solution (b) then 2 l
A statement has been added under Definition indicating
of reference solution (a).
that the monograph applies to riboflavin produced by
fermentation. This implies that the thresholds given
Drying : in a current of cold air.
under Related substances in the general monograph
Substances for pharmaceutical use (2034) do not apply to
this monograph.
System suitability : the chromatogram obtained with the
XXXX:0292
3rd application shows 2 clearly separated spots.
RIBOFLAVIN
with the test solution is similar in position and size to
the principal spot in the chromatogram obtained with
reference solution (a).
Riboflavinum
C. Dissolve about 1 mg in 100 ml of water R. The solution
has, by transmitted light, a pale greenish-yellow colour,
and, by reflected light, an intense yellowish-green
fluorescence which disappears on the addition of mineral
acids or alkalis.
TESTS
substance).
Dissolve 50.0 mg in 0.05 M sodium hydroxide free from
carbonate and dilute to 10.0 ml with the same alkaline
solution. Measure the optical rotation within 30 min of
C17H20N4O6
Mr 376.4 dissolution.
DEFINITION
Absorbance (2.2.25).
7,8-Dimethyl-10-[(2S,3S,4R)-2,3,4,5-tetrahydroxypentyl]Test solution. Dilute the final solution prepared for the assay
benzo[g]pteridine-2,4(3H,10H)-dione.
with an equal volume of water R.
This monograph applies to riboflavin produced by
Absorption maxima : at 223 nm, 267 nm, 373 nm and
fermentation.
444 nm.
Absorbance ratios :
A373/A267 = 0.31 to 0.33 ;
CHARACTERS
A444/A267 = 0.36 to 0.39.

Appearance : yellow or orange-yellow, crystalline powder.
Lumiflavin. Thin-layer chromatography (2.2.27).
Solubility : very slightly soluble in water, practically
Test solution (a). Suspend 25 mg in 10 ml of water R ;
insoluble in ethanol (96 per cent).
Solutions deteriorate on exposure to light, especially in the shake for 5 min and filter the suspension to remove the
undissolved material.
presence of alkali.
Test solution (b). Shake 25 mg of the substance to be
examined with 10.0 ml of methylene chloride R and filter
the suspension to remove the undissolved material.
IDENTIFICATION
Reference solution (a). Suspend 25 mg of riboflavin CRS in
A. Specific optical rotation (see Tests).
10 ml of water R ; shake for 5 min and filter the suspension
B. Examine the chromatogram obtained in the test for
to remove the undissolved material.
lumiflavin. Thin-layer chromatography (2.2.27).
Reference solution (b). Dissolve 25 mg of lumiflavin R
Test solution. Suspend 25 mg of the substance to be
examined in 10 ml of water R, shake for 5 min and filter in methylene chloride R and dilute to 50.0 ml with the
same solvent. Dilute 1.0 ml of the solution to 20.0 ml with
the suspension to remove the undissolved material.
methylene chloride R.
Reference solution (a). Suspend 25 mg of riboflavin CRS
in 10 ml of water R, shake for 5 min and filter the
to 100.0 ml with methylene chloride R.
suspension to remove the undissolved material.
Plate : TLC silica gel plate R (2-10 m)(29).
Reference solution (b). Dissolve 25 mg of lumiflavin R
(impurity A) in methylene chloride R and dilute to 50.0 ml Mobile phase : water R.
with the same solvent. Dilute 1.0 ml of this solution to
Application : as follows, dry in a current of cold air after
20.0 ml with methylene chloride R.
each individual application :
Plate : TLC silica gel plate R (2-10 m)(28).
1st application : 2 l of methylene chloride R followed
by 2 l of test solution (a),
Mobile phase : water R.
Application : as follows, drying in a current of cold air
2nd application : 2 l of methylene chloride R followed by
after each individual application :
2 l of reference solution (a),
(28) Precoated HPTLC plate silica gel, layer thickness 0.1 mm, Merck 1.12363 is suitable.
(29) Precoated HPTLC plate silica gel, layer thickness 0.1 mm, Merck 1.12363 is suitable.
289
Riboflavin
3rd application : 2 l of reference solution (b) followed by

2 l of reference solution (a),
4th application : 10 l of test solution (b),
5th application : 10 l of reference solution (c).
Drying : in a current of cold air.
System suitability : the chromatogram obtained with the 3rd
application shows 2 clearly separated spots.
Limit : in the chromatogram obtained with test solution (b) :
lumiflavin : any spot corresponding to lumiflavin is not
more intense than the principal spot in the chromatogram
obtained with reference solution (c) (0.025 per cent).
Prepare the solutions immediately before use and protect
them from light.

Mobile phase :
mobile phase A : phosphoric acid R, water R (1:1000 V/V) ;
Time
(min)
0-5
Mobile phase A
(per cent V/V)
90
Mobile phase B
(per cent V/V)
10
5 - 20
90 80
10 20
20 - 25
80
20
25 - 35
80 50
20 50
35 - 45
50
50
45 - 47
50 90
50 10
47 - 55
90
10

Injection : 10 l.
Relative retention with reference to riboflavin (retention
time = about 16 min) : impurity C = about 0.2 ;
impurity D = about 0.5 ; impurity A = about 1.4 ;
impurities A and B in the chromatogram obtained with
reference solution (c) ;
the chromatogram obtained with reference solution (b)
is similar to the chromatogram supplied with riboflavin
for peak identification CRS.
Limits :
impurity B = 1.4 ; impurity C = 2.3 ; impurity D = 1.4 ;
impurity A : not more than 0.25 times the area of the
1. impurity C 3. riboflavin
5. impurity B
impurities B, C, D : for each impurity, not more than
twice the area of the principal peak in the chromatogram
2. impurity D 4. impurity A
Figure 0292.-1. Chromatogram for the test for related
substances of riboflavin : solution of riboflavin spiked with
impurities A, B, C and D
Solution A : 13.6 g/l solution of sodium acetate R.
Test solution. With the aid of ultrasound, dissolve 120.0 mg
of the substance to be examined in 10.0 ml of 0.1 M sodium
(0.5 per cent) ;
hydroxide and dilute to 100.0 ml with solution A.
to 10.0 ml with solution A. Dilute 1.0 ml of this solution to
(0.05 per cent).
100.0 ml with solution A.
Reference solution (b). With the aid of ultrasound, dissolve on 1.000 g by drying in an oven at 100-105 C.
6.0 mg of riboflavin for peak identification CRS (containing
impurities C and D) in 0.05 ml of 0.1 M sodium hydroxide
on
the residue obtained in the test for loss on drying.
and dilute to 5.0 ml with solution A.
The following chromatogram is shown for information but
will not be published in the European Pharmacopoeia.
Reference solution (c). In order to prepare impurities A

and B in situ, dissolve 10 mg of the substance to be examined
in 1.0 ml of 0.5 M sodium hydroxide. Expose to daylight
for 1.5 h. Add 0.5 ml of acetic acid R and dilute to 100 ml
with water R.
Column :
size : l = 0.25 m, = 4.6 mm ;
ASSAY
Carry out the assay protected from light.
In a brown-glass 500 ml volumetric flask, suspend 65.0 mg
in 5 ml of water R ensuring that it is completely wetted and
dissolve in 5 ml of dilute sodium hydroxide solution R.
As soon as dissolution is complete, add 100 ml of water R
and 2.5 ml of glacial acetic acid R and dilute to 500.0 ml
(30) Symmetry C18, Hypersil C18, Kromasil C18 and Nucleosil C18 are suitable.
290
Sodium alendronate
with water R. Place 20.0 ml of this solution in a 200 ml

brown-glass volumetric flask, add 3.5 ml of a 14 g/l solution
of sodium acetate R and dilute to 200.0 ml with water R.
Measure the absorbance (2.2.25) at the absorption maximum It is proposed to replace the current TLC test for
at 444 nm.
4-aminobutanoic acid with an LC test for related
substances. Only comments on this test are requested.
Calculate the content of C17H20N4O6 taking the specific
XXXX:1564
absorbance to be 328.
STORAGE
SODIUM ALENDRONATE
Natrii alendronas
IMPURITIES
C4H12NNaO7P2,3H2O
A. 7,8,10-trimethylbenzo[g]pteridine-2,4(3H,10H)-dione
(lumiflavin),
B. 7,8-dimethylbenzo[g]pteridine-2,4(1H,3H)-dione,
C. 6,7-dimethyl-8-[(2S,3S,4R)-2,3,4,5-tetrahydroxypentyl]pteridine-2,4(3H,8H)-dione,
D. 8-(hydroxymethyl)-7-methyl-10-[(2S,3S,4R)-2,3,4,5tetrahydroxypentyl]benzo[g]pteridine-2,4(3H,10H)-dione.
Mr 325.1
DEFINITION
(4-Amino-1-hydroxybutylidene)bisphosphonic acid
monosodium salt trihydrate.
Content : 98.0 to 102.0 per cent (dried substance).
CHARACTERS
Solubility : soluble in water, very slightly soluble in
methanol, practically insoluble in methylene chloride.
IDENTIFICATION
Comparison : sodium alendronate CRS.
B. It gives reaction (a) of sodium (2.3.1).
TESTS
prepared from distilled water R and dilute to 50 ml with the
same solvent.
more intensely coloured than reference solution B7 or BY7
(2.2.2, Method II).
4-aminobutanoic acid. Examine by thin-layer
chromatography (2.2.27), using a TLC silica gel plate R.
solvent.
Reference solution (a). Dissolve 0.10 g of 4-aminobutanoic
acid R in water R and dilute to 200 ml with the same solvent.
to 10 ml with water R.
Apply to the plate 5 l of the test solution and 5 l of
reference solution (b). Allow the plate to dry in air. Develop
over a path of 15 cm using a mixture of 20 volumes of
water R, 20 volumes of glacial acetic acid R and 60 volumes
of butanol R. Dry the plate in a current of warm air. Spray
with ninhydrin solution R and heat at 100 C to 105 C for
15 min. Any spots corresponding to 4-aminobutanoic acid
in the chromatogram obtained with the test solution are not
with reference solution (b) (0.5 per cent).
291
Sodium alendronate

Solution A. Dissolve 29.4 g of sodium citrate R in water R
and dilute to 1.0 litres with the same solvent.
Buffer solution. Dissolve 5.88 g of sodium citrate R and
2.84 g of anhydrous disodium hydrogen phosphate R in
water R and dilute to 2.0 litres with the same solvent. Adjust
to pH 8 with phosphoric acid R and filter.
examined in solution A and dilute to 50.0 ml with solution A.
Take 5.0 ml of this solution and proceed as described
for reference solution (a), starting from to a 50 ml
polypropylene screw-cap centrifuge tube.
Reference solution (a). Dissolve 30 mg of the substance to
be examined and 30 mg of 4-aminobutanoic acid CRS in
solution A and dilute to 100.0 ml with solution A. Dilute
1.0 ml of this solution to 10.0 ml with solution A. Dilute
1.0 ml of this solution to 50.0 ml with solution A. Transfer
5.0 ml of this solution to a 50 ml polypropylene screw-cap
centrifuge tube containing 5 ml of a solution prepared by
dissolving 19.1 g of disodium tetraborate R in water R and
diluting to 1.0 litres with the same solvent. Add 5 ml of
acetonitrile R and 5 ml of a solution prepared immediately
before use by dissolving 200 mg of 9-fluorenylmethyl
chloroformate R in acetonitrile R and diluting to 50.0 ml
with the same solvent. Shake for 45 s and allow to stand
at room temperature for 30 min. Add 20 ml of methylene
chloride R and shake vigorously for 1 min. Centrifuge for
5-10 min and use a portion of the clear upper aqueous layer.
Reference solution (b). Take 5.0 ml of solution A and
proceed as described for reference solution (a), starting from
to a 50 ml polypropylene screw-cap centrifuge tube.
Column :
size : l = 0.25 m, = 4.1 mm ;
stationary phase : styrene-divinylbenzene copolymer R
(10 m)(31) ;
temperature : 45 C.
Mobile phase :
mobile phase A : acetonitrile R, buffer solution (3:17 V/V) ;
degas ;
mobile phase B : buffer solution, acetonitrile R (3:17 V/V);
degas ;
Time
(min)
0 - 15
Mobile phase A
(per cent V/V)
100 50
Mobile phase B
(per cent V/V)
0 50
15 - 25
50 0
50 100
25 - 27
0 100
100 0
27 - 32
100

Injection : 20 l.
Relative retention with reference to sodium alendronate
(retention time = about 7 min) : impurity A = about 1.8.
signal-to-noise ratio : not less than 3 for the peak due to
sodium alendronate ;
symmetry factor : not more than 2.0 for the peak due to
sodium alendronate.
Limits :

than the area of the peak due to sodium alendronate in
(0.10 per cent) ;
total : not more than 5 times the area of the peak due to
sodium alendronate in the chromatogram obtained with
disregard limit : 0.5 times the area of the peak due to
sodium alendronate in the chromatogram obtained with
reference solution (a) (0.05 per cent) ; disregard any peak
corresponding to those in the chromatogram obtained
with reference solution (b).
Phosphate and phosphite. Liquid chromatography (2.2.29).
solvent.
Reference solution (a). Dissolve 50.0 mg of sodium
alendronate CRS in water R and dilute to 25.0 ml with the
same solvent.
Reference solution (b). Dissolve 3.0 g of phosphoric acid R
in water R and dilute to 100.0 ml with the same solvent.
Dilute 1.0 ml of the solution to 100.0 ml with water R.
Reference solution (c). Dissolve 2.5 g of phosphorous acid R
in water R and dilute to 100.0 ml with the same solvent.
Dilute 1.0 ml of the solution to 100.0 ml with water R.
Reference solution (d). Mix 2.0 ml of reference solution (b)
and 2.0 ml of reference solution (c) and dilute to 50.0 ml
with water R.
Column :
size : l = 0.15 m, = 4.6 mm ;
stationary phase : anion exchange resin R1 (7 m) ;
temperature : 35 C.
Mobile phase : a solution of 0.2 ml of anhydrous formic
acid R in 1 litre of water R, adjusted to pH 3.5 with 2 M
sodium hydroxide solution.
Detection : differential refractometer.
Injection : 100 l.
Run time : twice the retention time of sodium alendronate.
Relative retention with reference to sodium alendronate
(retention time = about 16 min) : impurity B = about 1.3 ;
impurity C = about 1.6.
Limits :
impurities A, B : for each impurity, not more than the area
of the corresponding peak in the chromatogram obtained
with reference solution (d) (0.5 per cent).
1.0 g complies with test F. Prepare the reference solution
Loss on drying (2.2.32) : 16.1 per cent to 17.1 per cent,
determined on 1.000 g by drying in an oven at 140-145 C.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
phosphate and phosphite with the following modification.
Injection : test solution, reference solutions (a) and (d).
Calculate the percentage content of C4H12NNaO7P2 from the
declared content of sodium alendronate CRS.
(31) Hamilton PRP-1 is suitable.
292
Sodium hyaluronate
IMPURITIES
Specified impurities : A, B, C.
IDENTIFICATION
Comparison : Ph. Eur. reference spectrum of sodium
hyaluronate.
A. 4-aminobutanoic acid,
B. It gives reaction (a) of sodium (2.3.1).
B. phosphate,
TESTS
C. phosphite.
Reagents
9-Fluorenylmethyl chloroformate. C15H11ClO2. (Mr 258.7).
XXXXXXX. [28920-43-6].
Solution S. Weigh a quantity of the substance to be examined

equivalent to 0.10 g of the dried substance and add 30.0 ml
of a 9 g/l solution of sodium chloride R. Mix gently on a
shaker until dissolved (about 12 h).
Appearance of solution. Solution S is clear (2.2.1) and its
absorbance (2.2.25) at 600 nm is maximum 0.01.
pH (2.2.3) : 5.0 to 8.5.

It is proposed to introduce a test for heavy metals in view
of the usual dosage and routes of administration of the
product. In addition, it is now specified in the Assay that
reagent A must be cooled before use.
XXXX:1472
SODIUM HYALURONATE
Natrii hyaluronas
Dissolve the substance to be examined in carbon dioxide-free

water R to obtain a solution containing a quantity equivalent
to 5 mg of the dried substance per millilitre.
Intrinsic viscosity. Sodium hyaluronate is very hygroscopic
and must be protected from moisture during weighing.
Buffer solution (0.15 M sodium chloride in 0.01 M
phosphate buffer solution pH 7.0). Dissolve 0.78 g of
sodium dihydrogen phosphate R and 4.50 g of sodium
chloride R in water R and dilute to 500.0 ml with the same
solvent (solution A). Dissolve 1.79 g of disodium hydrogen
phosphate R and 4.50 g of sodium chloride R in water R
and dilute to 500.0 ml with the same solvent (solution B).
Mix solutions A and B until a pH of 7.0 is reached. Filter
through a sintered-glass filter (4).
Test solution (a). Weigh 0.200 g (m0p) (NOTE : this value
is only indicative and should be adjusted after an initial
measurement of the viscosity of test solution (a)) of the
substance to be examined and dilute with 50.0 g (m0s) of
buffer solution at 4 C. Mix the solution by shaking at 4 C
during 24 h. Weigh 5.00 g (m1p) of the solution and dilute
with 100.0 g (m1s) of buffer solution at 25 C. Mix this
solution by shaking for 20 min. Filter the solution through a
sintered-glass filter (100), and discard the first 10 ml.
(C14H20NNaO11)n
DEFINITION
Sodium salt of hyaluronic acid, a glycosaminoglycan
consisting of D-glucuronic acid and N-acetyl-D-glucosamine
disaccharide units.
Intrinsic viscosity : 90 per cent to 120 per cent of the value
stated on the label.
PRODUCTION
It is extracted from cocks combs or obtained by fermentation
from Streptococci, Lancefield Groups A and C. It is produced
by methods of manufacture designed to minimise or
eliminate infectious agents. When produced by fermentation
of gram-positive bacteria, the process must be shown to
reduce or eliminate pyrogenic or inflammatory components
of the cell wall.
CHARACTERS
Appearance : white or almost white, very hygroscopic
powder or fibrous aggregate.
Solubility : sparingly soluble or soluble in water, practically
insoluble in acetone and in anhydrous ethanol.
Test solution (b). Weigh 30.0 g (m2p) of test solution (a) and
dilute with 10.0 g (m2s) of buffer solution at 25 C. Mix this
sintered-glass filter (100) and discard the first 10 ml.
Test solution (c). Weigh 20.0 g (m3p) of test solution (a) and
Test solution (d). Weigh 10.0 g (m4p) of test solution (a) and
Determine the flow-times (2.2.9) for the buffer solution
(t0) and for the 4 test solutions (t1, t2, t3 and t4), at
25.00 0.03 C. Use an appropriate suspended level
viscometer (specifications : viscometer constant about
0.005 mm2/s2, kinematic viscosity of 1-5 mm2/s, internal
diameter of tube R 0.53 mm, volume of bulb C 5.6 ml,
internal diameter of tube N 2.8-3.2 mm) with a funnel-shaped
lower capillary end. Use the same viscometer for all
measurements ; measure all outflow times in triplicate. The
test is not valid unless the results do not differ by more than
0.35 per cent from the mean and if the flow time t1 is not less
than 1.6 and not more than 1.8 times t0. If this is not the
case, adjust the value of m0p and repeat the procedure.
293
Sodium hyaluronate
Calculation of the relative viscosities

Since the densities of the sodium hyaluronate solutions and
of the solvent are almost equal, the relative viscosities ri
(being r1, r2, r3 and r4) can be calculated from the ratio
of the flow times for the respective solutions ti (being t1,
t2, t3and t4) to the flow time of the solvent t0, but taking
into account the kinetic energy correction factor for the
capillary (B = 30 800 s3), using the following expression :
Calculation of the concentrations

Calculate the concentration c1 (expressed in kg/m3) of
sodium hyaluronate in test solution (a) using the following
expression:
percentage content of sodium hyaluronate as

determined under Assay ;
percentage loss on drying ;
25
1005 kg/m3 (density of the test solution at 25 C).

sodium hyaluronate in test solution (b) using the following
expression :
Calculate the concentration c3 (expressed in kg/m ) of

sodium hyaluronate in test solution (c) using the following
expression :

sodium hyaluronate in test solution (d) using the following
expression :
Sulphated glycosaminoglycans : maximum 1 per cent, if the

product is extracted from cocks combs.
Appropriate safety precautions are to be taken when
handling perchloric acid at elevated temperature.
Test solution. Introduce a quantity of the substance to be
examined equivalent to 50.0 mg of the dried substance into
a test-tube 150 mm long and 16 mm in internal diameter and
dissolve in 1.0 ml of perchloric acid R.
Reference solution. Dissolve 0.149 g of anhydrous sodium
sulphate R in water R and dilute to 100.0 ml with the same
solvent. Dilute 10.0 ml to 100.0 ml with water R. Evaporate
1.0 ml in a test-tube 150 mm long and 16 mm in internal
diameter in a heating block at 90-95 C, and dissolve the
residue in 1.0 ml of perchloric acid R.
Plug each test-tube with a piece of glass wool. Place the
test-tubes in a heating block or a silicone oil bath maintained
at 180 C and heat until clear, colourless solutions are
obtained (about 12 h). Remove the test-tubes and cool to
room temperature. Add to each test-tube 3.0 ml of a 33.3 g/l
solution of barium chloride R, cap and shake vigorously.
Allow the test-tubes to stand for 30 min. Shake each
test-tube once again, and determine the absorbance (2.2.25)
at 660 nm, using water R as a blank.
The absorbance obtained with the test solution is not greater
than the absorbance obtained with the reference solution.
Nucleic acids. The absorbance (2.2.25) of solution S at
260 nm is maximum 0.5.
Protein : maximum 0.3 per cent ; maximum 0.1 per cent, if
forms.
Test solution (a). Dissolve the substance to be examined in
water R to obtain a solution containing a quantity equivalent
to about 10 mg of the dried substance per millilitre.
Test solution (b). Mix equal volumes of test solution (a) and
water R.
Reference solutions. Prepare a 0.5 mg/ml stock solution
of bovine albumin R in water R. Prepare 5 dilutions of the
stock solution containing between 5 g/ml and 50 g/ml
of bovine albumin R.
Add 2.5 ml of freshly prepared cupri-tartaric solution R3
to test-tubes containing 2.5 ml of water R (blank), 2.5 ml
of the test solutions (a) or (b) or 2.5 ml of the reference
solutions. Mix after each addition. After about 10 min, add
to each test-tube 0.50 ml of a mixture of equal volumes of
phosphomolybdotungstic reagent R and water R prepared
immediately before use. Mix after each addition. After
30 min, measure the absorbance (2.2.25) of each solution
at 750 nm against the blank. From the calibration curve
obtained with the 5 reference solutions determine the
content of protein in the test solutions.
Chlorides (2.4.4) : maximum 0.5 per cent.
Dissolve 67 mg in 100 ml of water R.
Iron : maximum 80 80.0 ppm.
Calculation of the intrinsic viscosity

Calculate the intrinsic viscosity [] by linear least-squares
regression analysis using the Martin equation :
The decimal antilogarithm of the intercept is the intrinsic

viscosity expressed in m3/kg.
294
Atomic absorption spectrometry (2.2.23, Method II).

Test solution. Dissolve a quantity of the substance to be
examined equivalent to 0.25 g of the dried substance in 1 ml
of nitric acid R by heating on a water-bath. Cool and dilute
to 10.0 ml with water R.
Reference solutions. Prepare 2 reference solutions in the
same manner as the test solution, adding 1.0 ml and 2.0 ml
respectively of iron standard solution (10 ppm Fe) R to the
dissolved substance to be examined.
St. Johns wort dry extract, quantified
Source : iron hollow-cathode lamp using a transmission

band of 0.2 nm.
Calculate the percentage content of sodium hyaluronate

using the following expression :

Atomisation device : air-acetylene flame.
1.0 g complies with test F. Prepare the reference solution
using 2.0 ml of lead standard solution (10 ppm Pb) R.
cg
cs
on 0.500 g by drying at 100-110 C over diphosphorus
pentoxide R for 6 h.
Z
= mean of concentrations of D-glucuronic acid in

the test solutions, in milligrams per gram ;
= mean of concentrations of the substance to be
examined in the test solutions, in milligrams per
gram ;
= determined percentage content of C6H10O7 in
D-glucuronic acid R ;
= percentage loss on drying ;
Microbial contamination. Total viable aerobic count (2.6.12)

not more than 102 micro-organisms per gram. Use 1 g of the h
substance to be examined.
401.3 = relative molecular mass of the disaccharide
fragment ;
Bacterial endotoxins (2.6.14) : less than 0.5 IU/mg, if
194.1 = relative molecular mass of glucuronic acid.
forms without a further appropriate procedure for the
removal of bacterial endotoxins ; less than 0.05 IU/mg,
STORAGE
if intended for use in the manufacture of intra-ocular
In an airtight container, protected from light and humidity.
preparations or intra-articular preparations without a
If the substance is sterile, store in a sterile, airtight,
further appropriate procedure for the removal of bacterial
tamper-proof container.
endotoxins.
LABELLING
The label states :
ASSAY
the intrinsic viscosity ;
the origin of the substance ;
Determine the glucuronic acid content by reaction with
carbazole as described below.
the intended use of the substance ;
where applicable, that the substance is suitable for
Reagent A. Dissolve 0.95 g of disodium tetraborate R in
parenteral administration other than intra-articular
100.0 ml of sulphuric acid R.
administration ;
Reagent B. Dissolve 0.125 g of carbazole R in 100.0 ml of
where applicable, that the substance is suitable for
anhydrous ethanol R.
parenteral administration, including intra-articular
administration ;
Test solution. Prepare this solution in triplicate. Dissolve
where
applicable that the material is suitable for
0.170 g of the substance to be examined in water R and
intra-ocular
use.
dilute to 100.0 g with the same solvent. Dilute 10.0 g of this
solution to 200.0 g with water R.
Reference stock solution. Dissolve 0.100 g of D-glucuronic

acid R, previously dried to constant mass in vacuum over
diphosphorus pentoxide R (2.2.32), in water R and dilute to
100.0 g with the same solvent.
Reference solutions. Prepare 5 dilutions of the reference
stock solution containing between 6.5 g/g and 65 g/g of
D-glucuronic acid R.
Place 25 test-tubes, numbered 1 to 25, in iced water. Add
1.0 ml of the 5 reference solutions in triplicate to the
test-tubes 1 to 15 (reference tubes), 1.0 ml of the 3 test
solutions in triplicate to the test-tubes 16 to 24 (sample
tubes), and 1.0 ml of water R to test-tube 25 (blank). Add
to each test-tube 5.0 ml of freshly prepared reagent A,
previously cooled in iced water. Tightly close the test-tubes
with plastic caps, shake the contents, and place on a water
bath for exactly 15 min. Cool in iced water, and add to each
test tube 0.20 ml of reagent B. Recap the tubes, shake, and
put them again on a water-bath for exactly 15 min. Cool to
room temperature and measure the absorbance (2.2.25) of
the solutions at 530 nm, against the blank.
From the calibration curve obtained with the mean
absorbances read for each reference solution, determine
the mean concentrations of D-glucuronic acid in the test
solutions.
Reference: PA/PH/Exp. 13B/T (00) 20 ANP 2R

XXXX:1874
ST. JOHNS WORT DRY EXTRACT,

QUANTIFIED
Hyperici herbae
extractum siccum quantificatum
DEFINITION
Quantified dry extract obtained from St. Johns wort (1438).
Content :
total hypericins, expressed as hypericin (C30H16O8 ;
Mr 504.5) : 0.10 per cent to 0.30 per cent (dried extract) ;
flavonoids, expressed as rutin (C27H28O15 ; Mr 595.5) :
6.0 per cent to 12.0 per cent (dried extract) ;
hyperforin (C35H52O4 ; Mr 536.8), depending on the stated
content of hyperforin : maximum 0.2 per cent (dried
extract), greater than 0.2 per cent and less than 2.0 per
cent (dried extract), or 2.0 per cent to 6.0 per cent (dried
extract).
295
PRODUCTION
The extract is produced from the herbal drug by a suitable
procedure using ethanol (50-80 per cent V/V) or methanol
(50-80 per cent V/V).
CHARACTERS
Appearance : brownish-grey powder.
IDENTIFICATION
Thin-layer chromatography (2.2.27).
Test solution. Disperse 0.25 g of the extract to be examined
in 5 ml of methanol R.
Reference solution. Dissolve 5 mg of rutoside trihydrate CRS
(rutin) and 5 mg of hyperoside R in methanol R and dilute
to 5 ml with the same solvent.
Plate : TLC silica gel plate R.
Mobile phase : anhydrous formic acid R, water R, ethyl
acetate R (6:9:90 V/V/V).
Application : 10 l of the test solution and 5 l of the
reference solution, as bands of 10 mm.
Drying : at 100-105 C for 10 min.
Detection : spray with a 10 g/l solution of diphenylboric
acid aminoethyl ester R in methanol R and then with a
50 g/l solution of macrogol 400 R in methanol R. Examine
the plate after about 30 min in ultraviolet light at 365 nm.
Results : see below the sequence of zones present in the
chromatograms obtained with the reference solution and the
test solution. Furthermore, other fluorescent zones may be
present in the chromatogram obtained with the test solution.
Top of the plate
_______
A yellowish-orange fluorescent
zone
2 red fluorescent zones (hypericin
and pseudohypericin)
_______
_______
zone
zone
zone
_______
Hyperoside : a yellowish-orange
fluorescent zone
Rutin : a yellowish-orange
fluorescent zone
Reference solution
zone (hyperoside)
Yellow and blue possibly
superimposed fluorescent zones
zone (rutin)
Test solution
TESTS
Loss on drying (2.8.17) : maximum 5.0 per cent.
ASSAY
Total hypericins. Liquid chromatography (2.2.29).
Test solution. Dissolve 70.0 mg of the extract to be examined
in 25.0 ml of methanol R. Sonicate and centrifuge the
solution. Expose the solution to daylight for 2 h.
Reference solution. Place a quantity of St. Johns Wort
standardised dry extract CRS corresponding to 1.5 mg of
hypericin (mass of hypericin = m2) in a 20 ml volumetric
1. pseudohypericin
2. hypericin
Figure 1874.-1. Chromatogram for the assay of total hypericins : test solution
296
flask. Dissolve with 1 ml of pyridine R and dilute to 20.0 ml

with methanol R. Dilute 1.0 ml of this solution to 25.0 ml
with methanol R.
Column :
size : l = 0.15 m, = 4.6 mm ;
chromatography R (5 m) ;
temperature : 40 C.
Mobile phase : mix 39 volumes of ethyl acetate R, 41 volumes
of a 15.6 g/l solution of sodium dihydrogen phosphate R
adjusted to pH 2 with phosphoric acid R, and 160 volumes
of methanol R.
Injection: 20 l.
Elution order : pseudohypericin, hypericin.
System suitability : test solution :
hypericin and pseudohypericin.
Calculate the percentage content of total hypericins,

expressed as hypericin, using the following expression :
A1
A2
A3
m1
m2
20
area of the peak due to pseudohypericin in the

chromatogram obtained with the test solution ;
area of the peak due to hypericin in the
area of the peak due to hypericin in the
chromatogram obtained with the reference
solution ;
mass of the extract to be examined in the test
solution, in milligrams ;
mass of hypericin in the reference solution, in
milligrams ;
dilution factor.
1. rutin
3. isoquercitroside
5. quercetin
7. hyperforin
2. hyperoside
4. quercitroside
6. biapigenin
8. adhyperforin
Figure 1874.-2. Chromatogram for the assay of hyperforin and flavonoids : test solution
297
Hyperforin and flavonoids. Liquid chromatography (2.2.29). Calculate the percentage content of flavonoids, expressed as
rutin, using the following expression :
Carry out the assay protected from light.
Test solution. Dissolve 75.0 mg of the extract to be examined
in 20.0 ml of a mixture of 80 volumes of methanol R and
20 volumes of water R. Sonicate and centrifuge the solution.
Reference solution. Dissolve 2.0 mg of rutoside
trihydrate CRS in 20.0 ml of a mixture of 80 volumes of
methanol R and 20 volumes of water R.
m3
Column :
m4
A5
A6
A7
A8
A9
A10
A11
size : l = 0.15 m, = 4.6 mm ;

chromatography R (3 m).
Mobile phase :
mobile phase A : phosphoric acid R, acetonitrile R
(3:1000 V/V) ;
mobile phase B : phosphoric acid R, water R (3:1000 V/V) ;
Time
(min)
08
Mobile phase A
(per cent V/V)
18
Mobile phase B
(per cent V/V)
82
8 18
18 53
82 47
18 18.1
53 97
47 3
18.1 19
97
19 29
97
29 30
97 18
3 82
30 40
18
82
Detection : spectrophotometer at 360 nm, then at 275 nm

after the elution of biapigenin (about 22 min).

mass of rutoside trihydrate CRS in the reference
declared percentage content of rutoside
trihydrate CRS ;
area of the peak due to rutin in the chromatogram
obtained with the reference solution ;
obtained with the test solution ;
area of the peak due to hyperoside in the
area of the peak due to isoquercitroside in the
area of the peak due to quercitroside in the
area of the peak due to quercetin in the
area of the peak due to biapigenin in the
chromatogram obtained with the test solution.
LABELLING
The label states the content of hyperforin, as presented
under Definition.
Injection: 10 l.
Elution order : rutin, hyperoside, isoquercitroside,
quercitroside, quercetin, biapigenin, hyperforin,
adhyperforin.
System suitability : test solution :

resolution : minimum 2.0 between the peaks due to rutin A general update of this monograph is proposed, including
and hyperoside, and minimum 2.0 between the peaks due notably :
to hyperforin and adhyperforin.
the introduction of an LC test for related substances ;
Calculate the percentage content of hyperforin using the
the deletion of the streptodornase test since there is a
shortage of the International Standard and relevant
impurities may now be detected by the new LC method.
XXXX:0356
m3
m4
2.3
A4
A5
298
STREPTOKINASE BULK
CONCENTRATED SOLUTION
mass of rutoside trihydrate CRS in the reference
declared percentage content of rutoside
trihydrate CRS ;
factor of calculation of hyperforin relative to
rutin ;
area of the peak due to hyperforin in the
obtained with the reference solution.
Streptokinasi solutio ad praeparationem

concentrata
DEFINITION
Streptokinase bulk concentrated solution is a preparation
of a protein obtained from culture filtrates of certain
strains of haemolytic Streptococcus group C ; it has the
property of combining with human plasminogen to form
plasminogen activator. It may contain buffer salts and other
auxiliary substances. The potency is not less than 510 IU
per microgram of nitrogen.
PRODUCTION
If intended for use in the manufacture of parenteral
dosage forms, The method of manufacture is validated to
demonstrate that the product, if tested, would comply with
the following test.
Standard of streptodornase, to obtain a solution containing

20 IU of streptodornase activity per millilitre. The
equivalence in International Units of the International
Standard is stated by the World Health Organisation.
To each of 8 numbered centrifuge tubes, add 0.5 ml of a 1 g/l

solution of sodium deoxyribonucleate R in imidazole buffer
solution pH 6.5 R. To tube number 1 and tube number 2 add
0.25 ml of imidazole buffer solution pH 6.5 R, 0.25 ml of
the test solution and, immediately, 3.0 ml of perchloric acid
(25 g/l HClO4). Mix, centrifuge at about 3000 g for 5 min and
measure the absorbances (2.2.25) of the supernatant liquids
CHARACTERS
at 260 nm, using as the compensation liquid a mixture of
1.0 ml of imidazole buffer solution pH 6.5 R and 3.0 ml of
Appearance : clear, colourless liquid.
perchloric acid (25 g/l HClO4) (absorbances A1 and A2). To
the other 6 tubes (numbers 3 to 8) add 0.25 ml, 0.25 ml,
IDENTIFICATION
0.125 ml, 0.125 ml, 0 ml and 0 ml respectively of imidazole
buffer solution pH 6.5 R ; add to each tube 0.25 ml of the
A. Place 0.5 ml of citrated human plasma in a haemolysis
test solution and 0 ml, 0 ml, 0.125 ml, 0.125 ml, 0.25 ml
polystyrene tube maintained in a water-bath at 37 C.
Add 0.1 ml of a dilution of the preparation to be examined and 0.25 ml respectively of the reference solution. Mix the
containing 10 000 IU of streptokinase activity per millilitre contents of each tube and heat at 37 C for 15 min. To
each tube add 3.0 ml of perchloric acid (25 g/l HClO4), mix
in phosphate buffer solution pH 7.2 R and 0.1 ml of a
and centrifuge. Measure the absorbances (2.2.25) of the
solution of human thrombin R containing 20 IU/ml in
phosphate buffer solution pH 7.2 R. Mix immediately. A supernatant liquids at 260 nm using the compensation liquid
clot forms and lyses within 30 min. Repeat the procedure described above (absorbances A3 to A8). The absorbances
comply with the following requirement :
using citrated bovine plasma. The clot does not lyse
within 60 min.
Abnormal toxicity (2.6.9). Inject into each mouse a quantity
of the preparation to be examined (if necessary, dilute
with water for injections R) equivalent to 50 000 IU of
streptokinase activity in 0.5 ml, the injection lasting 15-20 s.
B. Dissolve 0.6 g of agar in 50.0 ml of barbital buffer

solution pH 8.6 R1, heating until a clear solution is
obtained. Use glass plates 50 mm square (transparency
mounts) free from traces of grease. Using a pipette, apply
to each plate 4 ml of the agar solution. Maintain the
plates horizontal. Allow to cool. Pierce a cavity 6 mm
in diameter in the centre of the agar and an appropriate
number of cavities (not exceeding 6) at distances of
11 mm from the central cavity. Remove the residual agar
from the cavities using a cannula connected to a vacuum
pump. Using pipettes graduated in microlitres, Perform
an immunochemical test using double immunodiffusion
techniques (2.7.1). Place in the central cavity about 80 l
of goat or rabbit antistreptokinase serum containing
about 10 000 units of antistreptokinase activity per
millilitre ; place in each of the surrounding cavities about
80 l of a dilution of the preparation to be examined
containing 125 000 IU of streptokinase activity per
millilitre. Allow the plates to stand in a humidified tank
for 24 h. Only one precipitation arc appears and it is well
defined and localised between the application point of
the serum and each cavity containing the solution of the
preparation to be examined.
TESTS
Streptolysin. In a haemolysis polystyrene tube, use a

quantity of the preparation to be examined equivalent to
500 000 IU of streptokinase activity and dilute to 0.5 ml with
a mixture of 1 volume of phosphate buffer solution pH 7.2 R
and 9 volumes of a 9 g/l solution of sodium chloride R. Add
0.4 ml of a 23 g/l solution of sodium thioglycollate R. Heat
in a water-bath at 37 C for 10 min. Add 0.1 ml of a solution
of a reference preparation of human antistreptolysin O
containing 5 IU/ml. Heat at 37 C for 5 min. Add 1 ml
of rabbit erythrocyte suspension R. Heat at 37 C for
30 min. Centrifuge at about 1000 g. In the same manner,
prepare a haemolysis tube in which the solution of the
preparation to be examined has been replaced by 0.5 ml of a
mixture of 1 volume of phosphate buffer solution pH 7.2 R
and 9 volumes of a 9 g/l solution of sodium chloride R.
Measure the absorbances (2.2.25) of the supernatant liquids
at 550 nm. The absorbance of the test solution is not more
than 50 per cent greater than that of the reference solution.
Test solution. Dilute the preparation to be examined

with water R to obtain a concentration of about 0.5-1 g/l,
Dilute the preparation to be examined in carbon dioxide-free depending on the chromatographic system used.
water R to obtain a solution containing 5000 IU at least
Reference solution. Dilute 1 volume of streptokinase for
1000 000 IU of streptokinase activity per millilitre.
system suitability CRS in 49 volumes of water R.
Streptodornase : maximum 10 IU of streptodornase activity
per 100 000 IU of streptokinase activity.
Column :
Test solution. Dilute the preparation to be examined in
size : l = 0.10 m, = 4.6 mm ;
imidazole buffer solution pH 6.5 R to obtain a solution
containing 150 000 IU of streptokinase activity per millilitre.
stationary phase : styrene-divinylbenzene copolymer R
Reference solution. Dissolve in imidazole buffer solution
(10 m) with a pore size of 200 nm(32) ;
pH 6.5 R a reference preparation of streptodornase,
calibrated in International Units against the International
temperature : 25 C.
pH (2.2.3) : 6.8 to 7.5.
(32) Poros R2 from Applied Biosystems is suitable.
299
Figure 0356.-1. Chromatogram for the test for related substances of streptokinase : reference solution
Mobile phase :
mobile phase A : trifluoroacetic acid R, water for
injections R (1:1000 V/V) ; degas ;
mobile phase B : trifluoroacetic acid R, acetonitrile for
chromatography R (1:1000 V/V) ; degas ;
Bacterial endotoxins (2.6.14) : less than 0.02 IU per 100 IU of

streptokinase activity, if intended for use in the manufacture
of parenteral dosage forms without a further appropriate
procedure for the removal of bacterial endotoxins.
ASSAY
Nitrogen (2.5.9).
Time
Mobile phase A
Mobile phase B
Potency
(min)
(per cent V/V)
(per cent V/V)
68
32
0-1
The potency of streptokinase is determined by comparing its
capacity to activate plasminogen to form plasmin with the
32 48
1-4
68 52
same capacity of a reference preparation of streptokinase
48
4-5
52
calibrated in International Units ; the formation of plasmin is
determined using a suitable chromogenic substrate.
0
5-7
100
The International Unit is the activity of a stated amount
68
32
7 - 10
of the International Standard for streptokinase. The
equivalence in International Units of the International
The above conditions may be modified to improve the
Standard is stated by the World Health Organisation.
separation efficiency of the chromatographic system.
Reference and test solutions
Prepare 2 independent series of 4 dilutions at least
3 dilutions of each of the substance to be examined
and of the reference preparation of streptokinase in
Injection : 20 l.
tris(hydroxymethyl)aminomethane sodium chloride buffer
solution pH 7.4 R1, in the linear range of the assay (a range
retention time : streptokinase = 2.3 min to 2.8 min ;
of 0.5-4.0 IU/ml has been found suitable). Prepare and
maintain all solutions at 37 C.
symmetry factor : maximum 1.9 for the peak due to
Substrate solution
streptokinase ;
the chromatogram obtained with the reference solution is Mix 1.0 ml of tris(hydroxymethyl)aminomethane buffer
similar to the chromatogram supplied with streptokinase solution pH 7.4 R with 1.0 ml of chromophore substrate R3.
Add 5 l of a 100 g/l solution of polysorbate 20 R. Keep at
for system suitability CRS.
37 C in a water-bath. Immediately before commencing the
Limit :
activation assay, add 45 l of a 1 mg/ml solution of human
plasminogen R.
total: maximum 5 per cent.
300
Method
Analyse each streptokinase dilution, maintained at 37 C, in
duplicate. Initiate the activation reaction by adding 60 l
of each dilution to 40 l of substrate solution. For blank
wells, use 60 l of tris(hydroxymethyl)aminomethane
sodium chloride buffer solution pH 7.4 R1 instead of the
reference and test solutions. Allow the reaction to proceed
at 37 C for 20 min and read the absorbance (2.2.25) at
405 nm. If a suitable thermostatted plate reader is available,
this may be used to monitor the reaction. Alternatively, it
may be necessary to stop the reaction after 20 min using
50 l of a 50 per cent V/V solution of glacial acetic acid R.
Best results are obtained when the absorbance for the
highest streptokinase concentration is between 0.1 and 0.2
(after blank subtraction). If necessary, adjust the time of
incubation in order to reach this range of absorbances.
Calculate the regression of the absorbance on log
concentrations of the solutions of the substance to be
examined and of the reference preparation of streptokinase
and calculate the potency of the substance to be examined
using the usual statistical methods for parallel-line assays.
The estimated potency is not less than 90 per cent and not
more than 111 per cent of the stated potency. The confidence
limits (P = 0.95) of the estimated potency are not less than
80 per cent and not more than 125 per cent of the stated
potency of the estimated potency.
STORAGE
In a sealed an airtight container, protected from light and at
a temperature of 20 C. If the substance is sterile, store in
a sterile, airtight, tamper-proof container.
LABELLING
The label states :
the number of International Units of streptokinase
activity per milligram, calculated with reference to the
dried preparation ;
the name and quantity of any added substance ;
where applicable, that the substance is free from bacterial
endotoxins ;
where applicable, that the substance is suitable for use in
the manufacture of parenteral dosage forms.

The TLC test for related substances has been replaced
by an LC test. The corresponding impurity section has
been introduced. Only comments on the test for related
substances are requested.
XXXX:0534
TRIMIPRAMINE MALEATE
Trimipramini maleas
C24H30N2O4
Mr 410.5
DEFINITION
(2RS)-3-(10,11-Dihydro-5H-dibenzo[b,f]azepin-5-yl)-N,N,2trimethylpropan-1-amine (Z)-but-2-enedioate.
CHARACTERS
Solubility : slightly soluble in water and in ethanol (96 per
cent).
IDENTIFICATION
First identification : A, C.
Second identification : A, B, D, E.
A. Melting point (2.2.14) : 140 C to 144 C.
B. Ultraviolet and visible absorption spectrophotometry
(2.2.25).
Test solution. Dissolve 40.0 mg in 0.01 M hydrochloric
acid and dilute to 100.0 ml with the same acid.
Dilute 5.0 ml of the solution to 100.0 ml with 0.01 M
hydrochloric acid.
Spectral range : 230-350 nm.
Absorption maximum : at 250 nm.
Shoulder : at 270 nm.
Specific absorbance at the absorption maximum : 205
to 235.
C. Infrared absorption spectrophotometry (2.2.24).
Comparison : trimipramine maleate CRS.
D. Thin-layer chromatography (2.2.27). Prepare the
solutions immediately before use.
same solvent. Dilute 1 ml of this solution to 20 ml with
methanol R.
Reference solution. Dissolve 25 mg of trimipramine
maleate CRS in methanol R and dilute to 10 ml with the
same solvent.
Plate : TLC silica gel G plate R.
Mobile phase : concentrated ammonia R, anhydrous
ethanol R, toluene R (0.7:10:90 V/V/V).
Application : 5 l.
Drying : in air for 15 min.
Detection : spray with a 5 g/l solution of potassium
dichromate R in a mixture of 1 volume of sulphuric acid R
and 4 volumes of water R and examine immediately.
with the test solution is similar in position, colour and
size to the principal spot in the chromatogram obtained
with the reference solution.
E. Thin-layer chromatography (2.2.27).
same solvent.
Reference solution. Dissolve 56 mg of maleic acid R in
methanol R and dilute to 10 ml with the same solvent.
Plate : TLC silica gel GF254 R.
Mobile phase : water R, anhydrous formic acid R,
di-isopropyl ether R (3:7:90 V/V/V).
Application : 5 l, as bands of 10 mm.
Drying : in a current of air for few minutes and then at
120 C for 10 min.
301
Results : the chromatogram obtained with the test

solution shows 2 zones : one is on the starting line and
the other is similar in position and size to the principal
zone in the chromatogram obtained with the reference
solution.
TESTS
Appearance of solution. The solution is not more intensely
coloured than reference solution BY5 (2.2.2, Method II).
Dissolve 2.5 g in chloroform R and dilute to 25 ml with the
same solvent.
(2.2.27), using silica gel G R as the coating substance.
examined in methanol R and dilute to 10 ml with the same
solvent. Prepare immediately before use.
with methanol R.
Reference solution (a). Dissolve 25 mg of trimipramine
maleate CRS in methanol R and dilute to 10 ml with the
same solvent. Prepare immediately before use.
to 10 ml with methanol R.
Reference solution (c). Dilute 1 ml of reference solution (a)
to 25 ml with methanol R.
Reference solution (d). Dissolve 10 mg of iminodibenzyl R
in methanol R and dilute to 100 ml with the same solvent.
Prepare immediately before use.
over a path of 15 cm using a mixture of 0.7 volumes
of concentrated ammonia R, 10 volumes of ethanol R
and 90 volumes of toluene R. Allow the plate to dry in
air for 15 min, spray with a 5 g/l solution of potassium
dichromate R in a mixture of 1 volume of sulphuric acid R
and 4 volumes of water R and examine immediately. In
the chromatogram obtained with test solution (a) : any
spot corresponding to iminodibenzyl is not more intense
than the spot in the chromatogram obtained with reference
solution (d) (0.2 per cent) ; any spot, apart from the principal
spot and the spot corresponding to iminodibenzyl, is not
with reference solution (b) (0.5 per cent) and at most
three such spots are more intense than the spot in the
chromatogram obtained with reference solution (c) (0.2 per
cent). Disregard any spot at the starting point.
Liquid chromatography (2.2.29). Carry out the test protected
from light.
Buffer solution pH 7.7. 2.64 g/l solution of ammonium
phosphate R in water for chromatography R. Adjust to
pH 7.7 with phosphoric acid R.
examined in the mobile phase and dilute to 100.0 ml with
the mobile phase.
Reference solution (a). Dissolve 5.0 mg of trimipramine
maleate for peak identification CRS (containing impurities
E and F) in the mobile phase and dilute to 10.0 ml with the
mobile phase.
100.0 ml with the mobile phase.
Reference solution (c). Dissolve 5.0 mg of trimipramine
maleate impurity mixture CRS (containing impurities A,
B, C and D) in the mobile phase and dilute to 10.0 ml with
the mobile phase. Dilute 1.0 ml of this solution to 100.0 ml
with the mobile phase.
Column :
size : l = 0.125 m, = 4.0 mm ;
Mobile phase : acetonitrile R1, buffer solution pH 7.7
(38:62 V/V).
Injection : 20 l.
Run time : 3 times the retention time of trimipramine.
1. impurity A
3. impurity C
5. trimipramine
7. impurity E
2. impurity B
4. impurity D
6. impurity F
8. impurity G
Figure 0534.-1. Chromatogram for the test for related substances of trimipramine maleate : solution of trimipramine
maleate spiked with 1 per cent of each impurity
(33) Kromasil C18 is suitable.
302
Relative retention with reference to trimipramine

impurity D = about 0.5 ; impurity F = about 1.4 ;
impurity E = about 1.5.
trimipramine and impurity F in the chromatogram
obtained with reference solution (a) ;
use the chromatogram supplied with trimipramine
maleate impurity mixture CRS and the chromatogram
obtained with reference solution (c) to identify the peaks
due to impurities A, B, C and D ;
use the chromatogram supplied with trimipramine
maleate for peak identification CRS and the
chromatogram obtained with reference solution (a) to
identify the peaks due to impurities E and F. Doubling of
the peak due to impurity E may be observed.
Limits :
the peak area of impurity F by 0.5 ;
impurities A, B, C, D : for each impurity, not more
(0.15 per cent) ;
impurity E : not more than 0.3 times the area of the
impurity F : not more than 0.2 times the area of the
(0.10 per cent) ;
sum of impurities other than E : maximum 0.50 per cent ;
(0.05 per cent).
on 1.0 g.

pharmaceutical use) : G.
A. [(2RS)-3-(10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)-2methylpropyl]dimethylamine N-oxide,
B. (2RS)-3-(10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)-N,2dimethylpropan-1-amine,
C. (2RS)-3-(5H-dibenzo[b,f]azepin-5-yl)-N,N,2-trimethylpropan-1-amine,
D. 3-(10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)-N,Ndimethylpropan-1-amine (imipramine),
ASSAY
Dissolve 0.3500 g in 50 ml of anhydrous acetic acid R.
Titrate with 0.1 M perchloric acid determining the end-point
E. (2RS)-N-[(2RS)-3-(10,11-dihydro-5H-dibenzo[b,f]azepinC24H30N2O4.
5-yl)-2-methylpropyl]-N,N,N,2-tetramethylpropane-1,3diamine,
STORAGE
IMPURITIES
Specified impurities : A, B, C, D, E, F.
impurities and/or by the general monograph Substances for F. 10,11-dihydro-5H-dibenzo[b,f]azepine,
303
Bacterial vaccines contain inactivated or live bacteria or

their antigenic components ; they are liquid preparations of
various degrees of opacity or they may be freeze-dried.
Bacterial toxoids are prepared from toxins by diminishing
their toxicity to a very low level or by completely eliminating
it by physical or chemical means whilst retaining adequate
immunising potency. The toxins are obtained from selected
strains of specified micro-organisms grown in suitable
media or are obtained by other suitable means, for example,
chemical synthesis.
G. 2-methyl-10,11-dihydro-5H-dibenzo[b,f]azepine.
The toxoids may be :
liquid,
precipitated with alum or another suitable agent,
purified and/or adsorbed on aluminium phosphate,
aluminium hydroxide, calcium phosphate or another
adsorbent prescribed in the monograph.
Bacterial
toxoids are clear or slightly opalescent liquids.
An addition has been made to the monograph concerning
Adsorbed
toxoids are suspensions or emulsions. Certain
the need in some cases to carry out an in-process test on
toxoids may be freeze-dried.
the inactivated harvest of clostridial vaccines to detect
contamination with other species of Clostridium produced Unless otherwise indicated, statements and requirements
given below for bacterial vaccines apply equally to bacterial
in the same facilities. Some species of Clostridium,
vaccines, bacterial toxoids and products containing a
notably Clostridium sordellii, are particularly resistant to
combination of bacterial cells and toxoid.
inactivation and are not detected by the general sterility
test. Test conditions suitable for Clostridium sordellii are
VIRAL VACCINES
indicated.
Viral vaccines are prepared by growth in suitable cell cultures
XXXX:0062 (5.2.4), in tissues, in micro-organisms, in fertilised eggs or,
where no other possibility is available, in live animals, or by
other suitable means. The strain of virus used may have
VACCINES FOR VETERINARY USE
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 Live viral vaccines are prepared from viruses of attenuated
virulence or of natural low virulence for the target species.
is the subject of a monograph in the Pharmacopoeia, the
Inactivated viral vaccines are treated by a validated
provisions of that monograph apply to that component,
procedure for inactivation of the virus and may be purified
modified where necessary as indicated (see Tests (Safety)
and concentrated.
below, Evaluation of safety of veterinary vaccines (5.2.6)
and Evaluation of efficacy of veterinary vaccines (5.2.7)).
VECTOR VACCINES
Vector vaccines are liquid or freeze-dried preparations of one
DEFINITION
or more types of live micro-organisms (bacteria or viruses)
Vaccines for veterinary use are preparations containing
that are non-pathogenic or have low pathogenicity for the
antigenic substances and are administered for the purpose
target species and in which have been inserted one or more
of inducing a specific and active immunity against disease
genes encoding antigens that stimulate an immune response
provoked by bacteria, toxins, viruses, fungi or parasites. The protective against other microorganisms.
vaccines, live or inactivated, confer active immunity that
may be transferred passively via maternal antibodies against PRODUCTION
the immunogens they contain and sometimes also against
The methods of preparation, which vary according to the
antigenically related organisms. Vaccines may contain
type of vaccine, are such as to maintain the identity and
bacteria, toxins, viruses or fungi, living or inactivated,
immunogenicity of the antigen and to ensure freedom from
parasites, or antigenic fractions or substances produced by
contamination with extraneous agents.
these organisms and rendered harmless whilst retaining
Substances of animal origin used in the production of
all or part of their antigenic properties ; vaccines may also
vaccines for veterinary use comply with the requirements of
contain combinations of these constituents. The antigens
chapter 5.2.5. Other substances used in the preparation of
may be produced by recombinant DNA technology. Suitable vaccines for veterinary use comply with requirements of the
adjuvants may be included to enhance the immunising
Pharmacopoeia (where a relevant monograph exists) and
properties of the vaccines.
are prepared in a manner that avoids contamination of the
Terminology used in monographs on vaccines for veterinary vaccine.
use is defined in chapter 5.2.1.
Clostridial vaccines. Where a clostridial vaccine is produced
in the same facilities as other clostridial vaccines, notably
BACTERIAL VACCINES AND BACTERIAL TOXOIDS
Clostridium sordellii vaccine, a test on the inactivated
Bacterial vaccines and bacterial toxoids are prepared from
cultures grown on suitable solid or liquid media, or by other harvest or the combined inactivated harvests for residual
live contaminating clostridia may be necessary since such
suitable means ; the requirements of this section do not
contaminants may not be detected by the test for sterility
apply to bacterial vaccines prepared in cell cultures or in
(2.6.1) because of the incubation time and media used.
live animals. The strain of bacterium used may have been
modified by genetic engineering. The identity, antigenic
For C. sordellii, sulphite polymyxin sulfadiazine agar (SPS
potency and purity of each bacterial culture used is carefully agar) has been found suitable. A part of the test sample
controlled.
is heated at 80 C for 10 min to allow germination of
304
endospores. The non-heated and heated portions of the

test sample are diluted 1:10-1:20 before inoculation. The
inoculated medium is incubated at 37 C for 60 days.
SUBSTRATES FOR PRODUCTION
Cell cultures used in the production of vaccines for veterinary
use comply with the requirements of chapter 5.2.4.
Where a monograph refers to chicken flocks free from
specified pathogens (SPF), these flocks comply with the
requirements prescribed in chapter 5.2.2.
For production of inactivated vaccines, where vaccine
organisms are grown in poultry embryos, such embryos
are derived either from SPF flocks (5.2.2) or from healthy
non-SPF flocks free from the presence of certain agents and
their antibodies, as specified in the monograph. It may be
necessary to demonstrate that the inactivation process is
effective against specified potential contaminants. For the
production of a master seed lot and for all passages of a
micro-organism up to and including the working seed lot,
eggs from SPF flocks (5.2.2) are used.
Where it is unavoidable to use animals or animal tissues in
the production of veterinary vaccines, such animals shall be
free from specified pathogens, as appropriate to the source
species and the target animal for the vaccine.
MEDIA
At least the qualitative composition must be recorded of
media used for seed culture preparation and for production.
The grade of each named ingredient is specified. Where
media or ingredients are claimed as proprietary, this
is indicated and an appropriate description recorded.
Ingredients that are derived from animals are specified as to
the source species and country of origin, and must comply
with the criteria described in chapter 5.2.5. Preparation
processes for media used, including sterilisation procedures,
are documented.
The addition of antibiotics during the manufacturing process
is normally restricted to cell culture fluids and other media,
egg inocula and material harvested from skin or other
tissues.
BACTERIAL SEED LOTS
General requirements. The genus and species (and varieties
where appropriate) of the bacteria used in the vaccine are
stated. Bacteria used in manufacture are handled in a
seed-lot system wherever possible. Each master seed lot
is tested as described below. A record of the origin, date
of isolation, passage history (including purification and
characterisation procedures) and storage conditions is
maintained for each master seed lot. Each master seed lot is
assigned a specific code for identification purposes.
Propagation. The minimum and maximum number of
subcultures of each master seed lot prior to the production
stage are specified. The methods used for the preparation
of seed cultures, preparation of suspensions for seeding,
techniques for inoculation of seeds, titre and concentration
of inocula and the media used, are documented. It shall be
demonstrated that the characteristics of the seed material
(for example, dissociation or antigenicity) are not changed
by these subcultures. The conditions under which each seed
lot is stored are documented.
Identity and purity. Each master seed lot is shown to
contain only the species and strain of bacterium stated. A
brief description of the method of identifying each strain by
biochemical, serological and morphological characteristics
and distinguishing it as far as possible from related strains is
recorded, as is also the method of determining the purity of
the strain. If the master seed lot is shown to contain living

organisms of any kind other than the species and strain
stated, then it is unsuitable for vaccine production.
VIRUS SEED LOTS
General requirements. Viruses used in manufacture are
handled in a seed-lot system. Each master seed lot is tested
as described below. A record of the origin, date of isolation,
passage history (including purification and characterisation
procedures) and storage conditions is maintained for each
seed lot. Each master seed lot is assigned a specific code
for identification purposes. Production of vaccine is not
normally undertaken using virus more than 5 passages
from the master seed lot. In the tests on the master seed lot
described below, the organisms used are not normally more
than 5 passages from the master seed lot at the start of the
tests, unless otherwise indicated.
Where the master seed lot is contained within a permanently
infected master cell seed, the following tests are carried out
on an appropriate volume of virus from disrupted master
cell seed. Where relevant tests have been carried out on
disrupted cells to validate the suitability of the master cell
seed, these tests need not be repeated.
Propagation. The master seed lot and all subsequent
passages are propagated on cells, on embryonated eggs
or in animals that have been shown to be suitable for
vaccine production (see above), and, where applicable, using
substances of animal origin that meet the requirements
prescribed in chapter 5.2.5.
Identification. A suitable method to identify the vaccine
strain and to distinguish it as far as possible from related
strains must be used.
Bacterial and fungal contamination. The master seed lot
complies with the test for sterility (2.6.1).
Mycoplasmas (2.6.7). The master seed lot complies with
the test for mycoplasmas (culture method and indicator cell
culture method).
Absence of extraneous viruses. Monographs may contain
requirements for freedom from extraneous agents, otherwise
the requirements stated below apply.
Preparations of monoclonal or polyclonal antibodies
containing high levels of neutralising antibody to the virus
of the seed lot are made on a batch basis, using antigen that
is not derived from any passage level of the virus isolate
giving rise to the master seed virus. Each batch of serum is
maintained at 56 C for 30 min to inactivate complement.
Each batch is shown to be free of antibodies to potential
contaminants of the seed virus and is shown to be free of
any non-specific inhibiting effects on the ability of viruses to
infect and propagate within cells (or eggs, where applicable).
If such a serum cannot be obtained, other methods are used
to remove or neutralise the seed virus specifically.
If the seed lot virus would interfere with the conduct and
sensitivity of a test for extraneous viruses, a sample of the
master seed lot is treated with a minimum amount of the
monoclonal or polyclonal antibody so that the vaccine
virus is neutralised as far as possible or removed. The final
virus-serum mixture shall, if possible, contain at least the
virus content of 10 doses of vaccine per 0.1 ml for avian
vaccines and per millilitre for other vaccines. For avian
vaccines, the testing to be carried out on seed lots is given in
chapter 2.6.24. For mammalian vaccines, the seed lot or the
mixture of seed lot and antiserum is tested for freedom from
extraneous agents as follows.
The mixture is inoculated onto cultures of at least 70 cm2 of
the required cell types. The cultures may be inoculated at
any suitable stage of growth up to 70 per cent confluency.
305
At least 1 monolayer of each type must be retained as a

control. The cultures must be monitored daily for a week. At
the end of this period the cultures are freeze thawed 3 times,
centrifuged to remove cell debris and re-inoculated onto the
same cell type as above. This is repeated twice. The final
passage must produce sufficient cells in appropriate vessels
to carry out the tests below.
Cytopathic and haemadsorbing agents are tested for using
the methods described in the relevant sections on testing cell
cultures (5.2.4) and techniques such as immuno-fluorescence
are used for detection of specific contaminants for the tests
in cell cultures. The master seed lot is inoculated onto :
primary cells of the species of origin of the virus,
cells sensitive to viruses pathogenic for the species for
which the vaccine is intended,
cells sensitive to pestiviruses.
If the master seed lot is shown to contain living organisms
of any kind, other than the virus of the species and strain
stated, or foreign viral antigens, then it is unsuitable for
vaccine production.
INACTIVATION
Inactivated vaccines are subjected to a validated inactivation
procedure. The testing of the inactivation kinetics described
below is carried out once for a given production process.
The rest of this section applies to each production run.
When conducting tests for inactivation, it is essential to
take account of the possibility that under the conditions of
manufacture, organisms may be physically protected from
inactivant.
Inactivation kinetics. The inactivating agent and the
inactivation procedure shall be shown, under conditions
of manufacture, to inactivate the vaccine micro-organism.
Adequate data on inactivation kinetics shall be obtained.
Normally, the time required for inactivation shall be not more
than 67 per cent of the duration of the inactivation process.
Aziridine. If an aziridine compound is used as the
inactivating agent then it shall be shown that no inactivating
agent remains at the end of the inactivation procedure.
This may be accomplished by neutralising the inactivating
agent with thiosulphate and demonstrating residual
thiosulphate in the inactivated harvest at the completion of
the inactivation procedure.
Formaldehyde. If formaldehyde is used as the inactivating
agent, then a test for free formaldehyde is carried out as
prescribed under Tests.
Other inactivating agents. When other inactivation methods
are used, appropriate tests are carried out to demonstrate
that the inactivating agent has been removed or reduced to
an acceptable residual level.
Inactivation and/or detoxification testing. A test for
complete inactivation and/or detoxification is performed
immediately after the inactivation and/or detoxification
procedure and, if applicable, the neutralisation or removal
of the inactivating or detoxifying agent.
Bacterial vaccines. The test selected shall be appropriate
to the vaccine bacteria being used and shall consist of at
least 2 passages in production medium or, if solid medium
has been used for production, in a suitable liquid medium
or in the medium prescribed in the monograph. The
product complies with the test if no evidence of any live
micro-organism is observed.
Bacterial toxoids. The test selected shall be appropriate to
the toxin or toxins present and shall be the most sensitive
available.
306
Viral vaccines. The test selected shall be appropriate to

the vaccine virus being used and must consist of at least
2 passages in cells, embryonated eggs or, where no other
suitably sensitive method is available, in animals. The
quantity of cell samples, eggs or animals shall be sufficient
to ensure appropriate sensitivity of the test. For tests in cell
cultures, not less than 150 cm2 of cell culture monolayer is
inoculated with 1.0 ml of inactivated harvest. The product
complies with the test if no evidence of the presence of any
live virus or other micro-organism is observed.
CHOICE OF VACCINE COMPOSITION AND CHOICE OF
VACCINE STRAIN
For the choice of vaccine composition and choice of
vaccine strain, important aspects to be evaluated include
safety, efficacy and stability. General requirements for
evaluation of safety and efficacy are given in chapter 5.2.6
and chapter 5.2.7. These requirements may be made more
explicit or supplemented by the requirements of specific
monographs.
For live vaccines, a maximum virus titre or bacterial count
acceptable from the point of view of safety is established
during development studies. This is then used as the
maximum acceptable titre for each batch of vaccine at
release.
Potency and immunogenicity. The tests given under the
headings Potency and Immunogenicity in monographs serve
2 purposes :
the Potency section establishes by a well-controlled test
in experimental conditions, the minimum acceptable
vaccinating capacity for all vaccines within the scope of
the definition, which must be guaranteed throughout the
period of validity ;
well-controlled experimental studies are normally a part
of the overall demonstration of efficacy of a vaccine
(see chapter 5.2.7) ; the test referred to in the section
Immunogenicity (which is usually a cross-reference to
the Potency section) is suitable as a part of this testing.
For most vaccines, the tests cited under Potency or
Immunogenicity are not suitable for the routine testing of
batches.
For live vaccines, the minimum acceptable virus titre
or bacterial count that gives satisfactory results in the
Potency test and other efficacy studies is established during
development. For routine testing it must be demonstrated
for each batch that the titre or count at release is such that
at the end of the period of validity, in the light of stability
studies, the vaccine, stored in the recommended conditions,
will contain not less than the minimum acceptable virus titre
or bacterial count determined during development studies.
For inactivated vaccines, if the test described under Potency
is not used for routine testing, a batch potency test is
established during development. The aim of the batch
potency test is to ensure that each batch of vaccine would,
if tested, comply with the test described under Potency
or Immunogenicity. The acceptance criteria for the batch
potency test are therefore established by correlation with
the test described under Potency. Where a batch potency
test is described in a monograph, this is given as an example
of a test that is considered suitable, after establishment of
correlation with the potency test ; other test models can also
be used.
Route of administration. During development of a vaccine,
safety and immunogenicity are demonstrated for each route
of administration to be recommended. The following is a
non-exhaustive list of such routes of administration :
intramuscular,
subcutaneous,
intravenous,
ocular,
oral,
nasal,
foot-stab,
wing web,
intradermal,
intraperitoneal,
in ovo.
Methods of administration. During development of a
vaccine, safety and immunogenicity are demonstrated
for each method of administration to be recommended.
The following is a non-exhaustive list of such methods of
administration :
injection,
drinking water,
spray,
eye-drop,
scarification,
implantation,
immersion.
Categories of animal. Monographs may indicate that a given
test is to be carried out for each category of animal of the
target species for which the product is recommended or is to
be recommended. The following is a non-exhaustive list of
categories that are to be taken into account.
Mammals :
pregnant animals/non-pregnant animals,
animals raised primarily for breeding/animals raised
primarily for food production,
animals of the minimum age or size recommended for
vaccination.
Avian species :
birds raised primarily for egg production/birds raised
primarily for production of meat,
birds before point of lay/birds after onset of lay.
Fish :
broodstock fish/fish raised primarily for food
production.
Stability. Evidence of stability is obtained to justify the
proposed period of validity. This evidence takes the form of
the results of virus titrations, bacterial counts or potency
tests carried out at regular intervals until 3 months beyond
the end of the shelf life on not fewer than 3 representative
consecutive batches of vaccine kept under recommended
storage conditions together with results from studies of
moisture content (for freeze-dried products), physical tests
on the adjuvant, chemical tests on substances such as
the adjuvant constituents and preservatives and pH, as
appropriate.
Where applicable, studies on the stability of the reconstituted
vaccine are carried out, using the product reconstituted in
accordance with the proposed recommendations.
FINAL BULK VACCINE
The final bulk vaccine is prepared by combining one or
more batches of antigen that comply with all the relevant
requirements with any auxiliary substances, such as
adjuvants, stabilisers, antimicrobial preservatives and
diluents.
Antimicrobial preservatives. Antimicrobial preservatives

are used to prevent spoilage or adverse effects caused by
microbial contamination occurring during use of a vaccine
which is expected to be no longer than 10 h after first
broaching. Antimicrobial preservatives are not included in
freeze-dried products but, if justified, taking into account the
maximum recommended period of use after reconstitution,
they may be included in the diluent for multi-dose
freeze-dried products. For single-dose liquid preparations,
inclusion of antimicrobial preservatives is not acceptable
unless justified and authorised, but may be acceptable, for
example where the same vaccine is filled in single-dose and
multidose containers and is used in non-food-producing
species. For multidose liquid preparations, the need for
effective antimicrobial preservation is evaluated taking into
account likely contamination during use and the maximum
recommended period of use after broaching of the container.
During development studies the effectiveness of the
antimicrobial preservative throughout the period of validity
shall be demonstrated to the satisfaction of the competent
authority.
The efficacy of the antimicrobial preservative is evaluated as
described in chapter 5.1.3 and in addition samples are tested
at suitable intervals over the proposed in use shelf-life. If
neither the A criteria nor the B criteria can be met, then in
justified cases the following criteria are applied to vaccines
for veterinary use : bacteria, no increase from 24 h to 7 days,
3 log reduction at 14 days, no increase at 28 days ; fungi, no
increase at 14 days and 28 days.
Addition of antibiotics as antimicrobial preservative is
generally not acceptable.
Test for inactivation and/or detoxification. For inactivated
vaccines, where the auxiliary substances would interfere
with a test for inactivation and/or detoxification, a test
for inactivation or detoxification is carried out during
preparation of the final bulk, after the different batches of
antigen have been combined but before addition of auxiliary
substances ; the test for inactivation or detoxification may
then be omitted on the final bulk and the batch.
Where there is a risk of reversion to toxicity, the test
for detoxification performed at the latest stage of the
production process at which the sensitivity of the test is not
compromised (e.g. after the different batches of antigen
have been combined but before the addition of auxiliary
substances) is important to demonstrate a lack of reversion
to toxicity.
In-process tests. Certain tests may be carried out on the
final bulk vaccine rather than on the batch or batches
prepared from it ; such tests include those for antimicrobial
preservatives, free formaldehyde and the potency
determination for inactivated vaccines.
BATCH
Unless otherwise prescribed in the monograph, the
final bulk vaccine is distributed aseptically into sterile,
tamper-proof containers which are then closed so as to
exclude contamination.
Only a batch that complies with each of the requirements
given below under Identification, Tests and Potency or in
the relevant individual monograph may be released for use.
With the agreement of the competent authority, certain of
the batch tests may be omitted where in-process tests give
an equal or better guarantee that the batch would comply
or where alternative tests validated with respect to the
Pharmacopoeia method have been carried out.
307
The identification test can often be conveniently combined

with the batch potency test to avoid unnecessary use of
animals. For a given vaccine, a validated in vitro test can be
used to avoid the unnecessary use of animals.
It is recognised that, in accordance with General Notices
(1.1. General statements), for an established vaccine the
routine application of the safety test will be waived by the
competent authority in the interests of animal welfare when
a sufficient number of consecutive production batches
have been produced and found to comply with the test,
thus demonstrating consistency of the manufacturing
process. Significant changes to the manufacturing process
may require resumption of routine testing to re-establish
consistency. The number of consecutive batches to be tested
depends on a number of factors such as the type of vaccine,
the frequency of production of batches and experience with
the vaccine during development safety testing and during
application of the batch safety test. Without prejudice
to the decision of the competent authority in the light
of information available for a given vaccine, testing of
10 consecutive batches is likely to be sufficient for most
products. For products with an inherent safety risk, it may
be necessary to continue to conduct the safety test on each
batch.
Animal tests. In accordance with the provisions of the
European Convention for the Protection of Vertebrate
Animals Used for Experimental and Other Scientific
Purposes, tests must be carried out in such a way as to use
the minimum number of animals and to cause the least pain,
suffering, distress or lasting harm. The criteria for judging
tests in monographs must be applied in the light of this. For
example, if it is indicated that an animal is considered to
be positive, infected etc. when typical clinical signs occur
then as soon as it is clear that the result will not be affected
the animal in question shall be either humanely killed or
given suitable treatment to prevent unnecessary suffering.
In accordance with the General Notices, alternative test
methods may be used to demonstrate compliance with
the monograph and the use of such tests is particularly
encouraged when this leads to replacement or reduction of
animal use or reduction of suffering.
Physical tests. A vaccine with an oily adjuvant is tested for
viscosity by a suitable method and shown to be within the
limits set for the product. The stability of the emulsion shall
be demonstrated.
Chemical tests. Tests for the concentrations of appropriate
substances such as aluminium and preservatives are carried
out to show that these are within the limits set for the
product.
pH. The pH of liquid products and diluents is measured and
shown to be within the limits set for the product.
Water. Where applicable, the freeze-drying process is
checked by a determination of water and shown to be within
the limits set for the product.
IDENTIFICATION
For inactivated vaccines, the identification prescribed in
monographs is usually an antibody induction test since this
is applicable to all vaccines.
TESTS
The monographs also indicate tests to be carried out on
each particular vaccine.
All hen eggs, chickens and chicken cell cultures for use
in quality control tests shall be derived from an SPF flock
(5.2.2).
308
Formaldehyde (2.4.18 ; use Method B if sodium

metabisulphite has been used to neutralise excess
formaldehyde). Where formaldehyde has been used in the
preparation, the concentration of free formaldehyde is not
greater than 0.5 g/l, unless a higher amount has been shown
to be safe.
Phenol (2.5.15). When the vaccine contains phenol, the
concentration is not greater than 5 g/l.
Sterility (2.6.1). Where prescribed in the monograph,
vaccines comply with the test for sterility. Where the volume
of liquid in a container is greater than 100 ml, the method of
membrane filtration is used wherever possible. Where the
method of membrane filtration cannot be used, the method
of direct inoculation may be used. Where the volume of
liquid in each container is at least 20 ml, the minimum
volume to be used for each culture medium is 10 per cent
of the contents or 5 ml, whichever is less. The appropriate
number of items to be tested (2.6.1) is 1 per cent of the batch
with a minimum of 4 and a maximum of 10.
For avian live viral vaccines, for non-parenteral use only, the
requirement for sterility is usually replaced by requirements
for absence of pathogenic micro-organisms and for a
maximum of 1 non-pathogenic micro-organism per dose.
Extraneous agents. Monographs prescribe a set of measures
that taken together give an acceptable degree of assurance
that the final product does not contain infectious extraneous
agents. These measures include :
1) production within a seed-lot system and a cell-seed system,
wherever possible ;
2) extensive testing of seed lots and cell seed for extraneous
agents ;
3) requirements for SPF flocks used for providing substrates
for vaccine production ;
4) testing of substances of animal origin, which must,
wherever possible, undergo an inactivation procedure ;
5) for live vaccines, testing of the final product for infectious
extraneous agents ; such tests are less extensive than those
carried out at earlier stages because of the guarantees given
by in-process testing.
In cases of doubt, the tests intended for the seed lot of a
live vaccine may also be applied to the final product. If an
extraneous agent is found in such a test, the vaccine does
not comply the monograph.
Avian live viral vaccines comply with the tests for extraneous
agents in batches of finished products (2.6.25).
Mycoplasmas (2.6.7). Where prescribed in a monograph,
the vaccine complies with the test for mycoplasmas (culture
method).
Safety. In general, 2 doses of an inactivated vaccine and/or
10 doses of a live vaccine are injected by a recommended
route. It may be necessary to reduce the prescribed number
of doses under certain circumstances or amend the method
of re-constitution and injection, for example for a combined
vaccine, where it is difficult to reconstitute 10 doses of the
live component in 2 doses of the inactivated component.
The animals are observed for the longest period stated in
the monographs. No abnormal local or systemic reaction
occurs. Where several batches are prepared from the same
final bulk, the safety test is carried out on the first batch and
then omitted for further batches prepared from the same
final bulk.
During development studies, the type and degree of reactions
expected with the vaccine are defined in the light of safety
testing. This definition is then used as part of the operating
procedure for the batch safety test to evaluate acceptable
and unacceptable reactions.
The immune status of animals to be used for the safety

test is specified in the individual monograph. For most
monographs, one of the 3 following categories is specified :
1) the animals must be free from antibodies against the
virus/bacterium/toxin etc. contained in the vaccine,
2) the animals are preferably free from antibodies but
animals with a low level of antibody may be used as long as
the animals have not been vaccinated and the administration
of the vaccine does not cause an anamnestic response,
3) the animals must not have been vaccinated against the
disease the vaccine is intended to prevent.
As a general rule, category 1 is specified for live vaccines.
For other vaccines, category 2 is usually specified but where
most animals available for use in tests would comply with
category 1, this may be specified for inactivated vaccines also.
Category 3 is specified for some inactivated vaccines where
determination of antibodies prior to testing is unnecessary
or impractical. For poultry vaccines, as a general rule the
use of SPF birds is specified.
For avian vaccines, the safety test is generally carried out
using 10 SPF chickens (5.2.2), except that for vaccines not
recommended for use in chickens it is carried out using
10 birds of one of the species for which the vaccine is
recommended, the birds being free from antibodies against
the disease agent for which the vaccine is intended to provide
protection.
any contra-indications to the use of the product including

any required warning on the dangers of administration
of an overdose,
the doses recommended for different species.
Reference: PA/PH/Exp. 10C/T (04) 12 ANP

The test for impurities E, F and G lists Rf values for more
impurities than those limited by the test. This information
is provided to users of the monograph to assist with spot
identification on the TLC plates. Impurities A and B have
limited solubility in the conditions of the test for related
substances and are therefore quantified in the test for
impurities A, B, I and R (enantiometric purity). A test for
chloride is included because stoichiometry is essential to
the chemical stability of the substance.
XXXX:1768
VALACICLOVIR HYDROCHLORIDE
Valacicloviri hydrochloridum
POTENCY
See Choice of vaccine composition and choice of vaccine
strain under Production.
STORAGE
Store protected from light at a temperature of 5 3 C,
unless otherwise indicated. Liquid preparations are not to
be allowed to freeze, unless otherwise indicated.
LABELLING
The label states :
that the preparation is for veterinary use,
the volume of the preparation and the number of doses
in the container,
the route of administration,
the type or types of bacteria or viruses used and for live
vaccines the minimum and the maximum number of live
bacteria or the minimum and the maximum virus titre,
where applicable, for inactivated vaccines, the minimum
potency in International Units,
where applicable, the name and amount of antimicrobial
preservative or other substance added to the vaccine,
the name of any substance that may cause an adverse
reaction,
for freeze-dried vaccines :
the name or composition and the volume of the
reconstituting liquid to be added,
the period within which the vaccine is to be used after
reconstitution,
for vaccines with an oily adjuvant, that if the vaccine is
accidentally injected into man, urgent medical attention
is necessary,
the animal species for which the vaccine is intended,
the indications for the vaccine,
the instructions for use,
C13H21N6O4Cl
Mr 360.8
DEFINITION
2-[(2-Amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
L-valinate hydrochloride.
substance).
CHARACTERS
Appearance : white or almost white powder.
Solubility : freely soluble in water, slightly soluble in
anhydrous ethanol, practically insoluble in 1-octanol.
IDENTIFICATION
Comparison : valaciclovir hydrochloride CRS.
substance separately in the minimum volume of
anhydrous ethanol R and evaporate to dryness in a
dessicator, under high vacuum, over diphosphorus
pentoxide R. Record new spectra using the residues.
B. It gives reaction (a) of chlorides (2.3.1).
TESTS
Impurities E, F and G. Thin-layer chromatography (2.2.27).
Solvent mixture : water R, anhydrous ethanol R (5:95 V/V).
examined in 2 ml of water R and dilute to 5.0 ml with the
solvent mixture.
309
Reference solution (a). Dissolve 5.0 mg each of valaciclovir

impurity D CRS, valaciclovir impurity E CRS and
valaciclovir impurity G CRS, and 8.4 mg of valaciclovir
impurity F para-toluenesulphonate CRS, in 2 ml of water R
and 6 ml of the solvent mixture and dilute to 10 ml with the
solvent mixture.
Reference solution (c). Dilute 2.0 ml of reference solution (a)
Reference solution (d). Dilute 0.5 ml of reference solution (a)
Plate : TLC silica gel F254 plate R (2-10 m)(34).
Pretreatment: wash the plate with methanol R until the
solvent front has migrated over at least 80 per cent of the
plate ; allow the plate to dry.
Mobile phase : concentrated ammonia R, tetrahydrofuran R,
methanol R, methylene chloride R (3:12:34:54 V/V/V/V).
Use freshly prepared mobile phase.
Application : 4 l of test solution (a) and reference
solutions (b), (c) and (d).
Development : over 80 per cent of the plate.
Drying : in a current of air.
Detection : examine in ultraviolet light at 254 nm for
impurities E and G ; spray with a 0.1 g/l solution of
fluorescamine R in ethylene chloride R and examine in
ultraviolet light at 365 nm for impurity F.
Rf value : impurity A = about 0.00 ; impurity B = about 0.20 ;
valaciclovir = about 0.34 ; impurity C = about 0.51 ;
impurity D = about 0.58 ; impurity E = about 0.68 ;
impurity F = about 0.75 ; impurity G = about 0.79. Impurity C
is masked by the leading edge of the spot due to valaciclovir.
Impurities F and G may co-elute, but this does not adversely
affect their quantification.
System suitability : the chromatograms obtained with

reference solutions (b), (c) and (d) each show 3 clearly
separated spots when examined under ultraviolet light at
254 nm, due to impurities D, E and G.
Limits :
impurity E : any spot due to impurity E is not more
intense than the corresponding spot in the chromatogram
obtained with reference solution (c) (0.2 per cent) ;
impurity F : any spot due to impurity F is not more
impurity G : any spot due to impurity G is not more
obtained with reference solution (d) (0.05 per cent).
Solvent mixture A : water R, anhydrous ethanol R
(5:95 V/V).
Solvent mixture B : solvent mixture A, water R (20:80 V/V).
examined in solvent mixture B and dilute to 100.0 ml with
solvent mixture B.
Reference solution (a). Dissolve 5 mg of valaciclovir for
peak identification CRS (containing impurities A, B, C, D,
H, I, J, M, N and P) in solvent mixture B and dilute to 10 ml
with solvent mixture B.
to 100.0 ml with solvent mixture B. Dilute 1.0 ml of this
solution to 20.0 ml with solvent mixture B.
Column :
size : l = 0.25 m, = 4.6 mm ;
stationary phase : phenylhexylsilyl silica gel for
chromatography R(35) ;
temperature : 15 C.
1. impurity A
4. impurity C
7. impurity K
10. impurity N
2. impurity B
5. impurity D
8. impurity L
11. impurity O
3. valaciclovir
6. impurity J
9. impurity M
12. impurity P
Figure 1768.-1. Chromatogram for the test for related substances of valaciclovir hydrochloride spiked with
impurities A, B, C, D, J, K, L, M, N, O, P
(34) Merck silica gel 60F254 HPTLC is suitable. 20 cm plates are preferable, but 10 cm plates are suitable too.
(35) Luna Phenyl Hexyl 5 m is suitable.
310
1. impurity B
3. valaciclovir
5. impurity I
7. impurity J
2. impurity H
4. impurity C
6. impurity D
8. impurity M
Figure 1768.-2. Chromatogram for the test for related substances of valaciclovir hydrochloride spiked with
impurities B, C, D, H, I, J, M
Mobile phase :
mobile phase A : trifluoroacetic acid R, water R
(3:1000 V/V) ;
mobile phase B : trifluoroacetic acid R, methanol R2
(3:1000 V/V) ;
Time
(min)
0-5
Mobile phase A
(per cent V/V)
90
Mobile phase B
(per cent V/V)
10
5 - 35
90 60
10 40
35 - 35.01
60 90
40 10
35.01 - 45
90
10

Injection: 10 l.
supplied with valaciclovir for peak identification CRS and
the chromatogram obtained with reference solution (a) to
identify the peaks due to impurities A, B, C, D, H, I, J, M, N
and P.
Relative retention with reference to valaciclovir (retention
impurity B = about 0.43 ; impurity H = about 0.54 ;
impurity C = about 1.06 ; impurity I = about 1.09 ;
impurity D = about 1.16 ; impurity J = about 1.30 ;
impurity M = about 1.60 ; impurity N = about 1.68 ;
impurity P = about 2.00 ; impurity Q = about 2.12.
valaciclovir and impurity C, and minimum 1.5 between
the peaks due to impurities C and I ;
the chromatogram obtained is similar to the
chromatogram supplied with valaciclovir for peak
identification CRS.
Limits :
impurity H : maximum 0.1 per cent ;
impurity C : maximum 0.3 per cent ;
impurity D : maximum 0.5 per cent ;
impurity J : maximum 0.1 per cent ;
impurity M : maximum 1.5 per cent ;

impurity N : maximum 0.2 per cent ;
impurity P : maximum 0.3 per cent ;
impurity Q : maximum 0.1 per cent ;
unspecified impurities : for each impurity, maximum
0.10 per cent ;
disregard limit : the area of the principal peak in the
(0.05 per cent) ; disregard the peaks due to impurities A,
B and I ;
total : see test for impurities A, B, I and R (enantiomeric
purity).
Impurities A, B, I and R (enantiomeric purity). Liquid
chromatography (2.2.29) : use the normalisation procedure.
examined in a 0.5 per cent (V/V) solution of hydrochloric
acid R and dilute to 100.0 ml with the same solution.
Reference solution. Dissolve 0.050 g of valaciclovir
hydrochloride CRS (containing impurities A, B, I and R) in
a 0.5 per cent (V/V) solution of hydrochloric acid R and
dilute to 100.0 ml with the same solution.
Column :
size : l = 0.15 m, = 4.0 mm ;
stationary phase : crown-ether silica gel for
chromatography R(36) ;
temperature : 10 C.
Mobile phase : perchloric acid R, methanol R, water R
(0.5:5:95 V/V/V).
Injection : 10 l.
Run time : 1.5 times the retention time of valaciclovir.
with valaciclovir hydrochloride CRS and the chromatogram
obtained with the reference solution to identify the peaks
due to impurities A, B, I and R.
Relative retention with reference to valaciclovir (retention
time = about 21 min) : impurities A and B = about 0.2 ;
impurity I = about 0.4 ; impurity R = about 0.6 ;
impurity D = about 0.7 ; impurity M = about 1.3.
(36) Daicel CrownPak CR (+) (5 m) is suitable.
311
1. impurities A et B
3. impurity R
5. valaciclovir
2. impurity I
4. impurity D
6. impurrity M
Figure 1768.-3. Chromatogram for the test for impurities A, B, I and R (enantiomeric purity) of valaciclovir
hydrochloride
impurity R and valaciclovir.
Limits :
the peak area of impurities A and B by 0.66 ;
sum of impurities A and B : maximum 2.0 per cent ;
impurity I : maximum 0.2 per cent ;
impurity R : maximum 3.0 per cent ;
total: from the tests for related substances and
impurities A, B, I and R (enantiomeric purity) : maximum
5.0 per cent.
Chloride : 9.4 to 9.9 per cent (anhydrous and solvent-free
substance).
Dissolve 350.0 mg in 100 ml of water R and add 4 drops of
nitric acid R. Carry out a potentiometric titration (2.2.20),
using 0.1 M silver nitrate. Use a glass indicator electrode and
a silver reference electrode or a combined silver electrode.
Discard the result from the first titration, which is used to
condition the electrodes. Carry out a blank titration.
1 ml of 0.1 M silver nitrate is equivalent to 3.5432 mg of Cl.
Palladium : maximum 10.0 ppm.
Atomic emission spectrometry (2.2.22, Method I).
Test solution. Dissolve 100 mg in a 2 per cent (V/V) solution
of hydrochloric acid R in dimethyl sulphoxide R and dilute
to 10.0 ml with the same solvent.
Reference solutions. Prepare the reference solutions using a
solution containing 1000 g of Pd per millilitre, diluted as
necessary with a 2 per cent V/V solution of hydrochloric
acid R in dimethyl sulphoxide R.
Dissolve 2.0 g in water R and dilute to 20 ml with the same
solvent. 12 ml of the solution complies with test A. Prepare
the reference solution using 10 ml of lead standard solution
(2 ppm) Pb R.
312
Water (2.5.12) : maximum 2.0 per cent, determined on

0.250 g.
on 2.0 g.
ASSAY
Liquid chromatography (2.2.29) as described in the test for
impurities A, B, I and R (enantiomeric purity).
Injection : test solution and reference solution.
Calculate the percentage content of C13H21N6O4Cl from the
declared content of valaciclovir hydrochloride CRS.
IMPURITIES
Specified impurities : A, B, C, D, E, F, G, H, I, J, M, N, P,
Q, R.
pharmaceutical use) : K, L, O.
A. 2-amino-1,9-dihydro-6H-purin-6-one (guanine),
B. 2-amino-9-[(2-hydroxyethoxy)methyl]-1,9-dihydro-6Hpurin-6-one,
C. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
N-methyl-L-valinate,
J. 2-[[(2-amino-6-oxo-1,6-dihydro-9H-purin-9yl)methyl]oxy]ethyl isoleucinate,
D. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
N-ethyl-L-valinate,
K. 9-[[(2-hydroxyethyl)oxy]methyl]-2-[[[(6-oxo-6,9-dihydro1H-purin-2-yl)amino]methyl]amino]-1,9-dihydro-6H-purin6-one,
L. 2,2-(methanediyldiimino)bis[9-[[(2-hydroxyethyl)oxy]methyl]-1,9-dihydro-6H-purin-6-one],
E. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
N-[(benzyloxy)carbonyl]-L-valinate,
F. 2-hydroxyethyl L-valinate,
G. N,N-dimethylpyridin-4-amine,
M. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
N-formyl-L-valinate,
N. 2-[[6-oxo-2-[[[(6-oxo-6,9-dihydro-1H-purin-2yl)amino]methyl]amino]-1,6-dihydro-9H-purin-9yl]methoxy]ethyl L-valinate,
H. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
L-alaninate,
I. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
acetate,
O. 2-[[2-[[[[9-[(2-hydroxyethoxy)methyl]-6-oxo-6,9-dihydro1H-purin-2-yl]amino]methyl]amino]-6-oxo-1,6-dihydro-9Hpurin-9-yl]methoxy]ethyl L-valinate,
313
Q. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9yl)methoxy]ethyl N-[[(6-oxo-6,9-dihydro-1H-purin2-yl)amino]methyl]valinate,
P. 2-[[2-[[[[9-[[2-[[(2S)-2-amino-3-methyl-1-methylenebutyl]oxy]ethoxy]methyl]-6-oxo-6,9-dihydro-1H-purin2-yl]amino]methyl]amino]-6-oxo-1,6-dihydro-9H-purin9-yl]methoxy]ethyl L-valinate,
R. 2-[(2-amino-6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy]ethyl
D-valinate.
Reagents
Crown-ether silica gel for chromatography. XXXXXXX.
Stationary phase for liquid chromatography.
Crown ether coated on silica gel.
314
Eucalyptus leaf
Illustrations of Powdered Drugs

in Herbal Monographs
It has been decided to progressively introduce illustrations of powdered drugs into herbal monographs in order to
complement the corresponding identification section (usually Identification B). These drawings will be published
in Pharmeuropa as they become available.

This monograph is currently published in the 5th Edition.
th
XXXX:1396 This monograph is currently published in the 5 Edition.
XXXX:1320
BOLDO LEAF
Boldi folium
EUCALYPTUS LEAF
Eucalypti folium
A. Fragment of the lamina, in

surface view, showing the upper
epidermis, hypodermis with
thickened and beaded walls and
palisade parenchyma
B. Lower epidermis with stomata
surrounded by 4 to 7 subsidiary
cells
C. Unicellular covering trichome,
solitary
D. Unicellular covering trichomes,
stellate clustered
E. Fragment of the lamina, in

transverse section, showing the
upper epidermis, hypodermis,
palisade parenchyma and spongy
parenchyma containing oil cells (G)
F. Spongy parenchyma containing
fine needle-shaped crystals and oil
cells (G)
G. Oil cells
H. Vascular tissue
Figure 1396.-1. Illustration of powdered herbal drug of

boldo leaf (see Identification B)
A. Thick-walled epidermal cells and

anomocytic stomata, in surface view
B. Epidermis covered by a thick cuticle,
in transverse section
C. Palisade parenchyma
E. Schizogenous oil gland

F. Vascular tissue
G. Fibres
D. Cells containing prisms and cluster

crystals of calcium oxalate
Figure 1320.-1. Illustration of powdered herbal drug of

eucalyptus leaf (see Identification B)
315
316
Copovidone
International Harmonisation
This section contains proposals for monographs and general texts, new or revised, elaborated under the international
harmonisation procedure (see chapter 5.8 of the European Pharmacopoeia). After these texts have undergone the
harmonisation procedure and have been adopted, they will be included in the European Pharmacopoeia and the
pharmacopoeias of the United States and Japan.
The draft harmonised texts are published for comment (stage 4: official public enquiry in the forum of each of the three
pharmacopoeias). You may send your comments through the appropriate national pharmacopoeia authority at the
address listed on the inside back cover of this issue. Readers whose country is not a signatory state of the European
Pharmacopoeia Convention can send their comments directly to the European Directorate for the Quality of Medicines
(see address of the EDQM on the cover of this issue). To facilitate the processing of comments received by the Secretariats
of the national authorities and the EDQM please mention in any correspondence the PA/PH reference number at
the beginning of each text. If you are requesting a change in the limits or are proposing other methods of analysis,
please support your proposal by providing appropriate analytical data obtained on a significant number of samples
and the results of a comparative study between the official method and the proposed method. Comments sent before
30 June 2006 will be considered for the preparation of the final version of the harmonised texts.
We wish to emphasise that these draft texts have not yet been adopted by the European Pharmacopoeia Commission
and therefore cannot be considered to be official texts.
It should be noted that for monographs, a version drafted in the European Pharmacopoeia style is usually published at
the same time in the section on Draft monographs and general texts for comments.
Description
Copovidone occurs as a white to slightly yellowish powder or
flakes. It is very soluble in methanol and in ethanol, freely
Stage 4
soluble in water. It is hygroscopic.
The JP is the coordinating pharmacopoeia for this
Identification
monograph.
Determine the infrared absorption spectrum of Copovidone,
The draft for harmonisation presented below is essentially previously dried at 105C for 3 hours, as directed in
for information. In the section on monographs for
the potassium bromide disk method under the Infrared
comment of this issue of Pharmeuropa, a revision
Spectrophotometry, and compare the spectrum with
proposal for the European Pharmacopoeia monograph
the Reference Spectrum or the spectrum of Copovidone
on copovidone is published. This shows how the stage 4
Reference Standard previously dried at 105C for 3 hours :
draft would affect the European monograph. Comments
both spectra exhibit similar intensities of absorption at the
are invited on the revision proposal (rather than on the
same wave numbers.
JP draft).
K-value
XXXX:0891
Weigh exactly an amount of Copovidone, equivalent to
1.000 g, calculated on the dried basis, and dissolve in water
COPOVIDONE
to make exactly 100 ml, allow to stand for 60 minutes,
and use this solution as the sample solution. Perform the
test with the sample solution and with water at 25C as
directed in Method 1 under the Viscosity Determination, and
calculate the K-value by the following formula. The K-value
of Copovidone is not less than 90.0% and not more than
110.0% of the nominal K-value.
(C6H9NO)n, (C4H6O2)m
Mr (111.14)n + (86.09)m
Definition
Copolymer of 1-ethenylpyrrolidin-2-one and ethenyl acetate
(Poly[(2-oxopyrrolidin-1-yl)ethylene-co-(1-acetoxyethylene)])
[25086-89-9]
Copovidone is a copolymer of 1-vinyl-2-pyrrolidone and vinyl
acetate at the ratio by weight of 3:2.
It, calculated on the dried basis, contains not less than 7.0%
and not more than 8.0% of nitrogen (N : 14.01), and not
less than 35.3% and not more than 42.0% of vinyl acetate
(C4H6O2 ; 86.09).
Labelling
Label it to indicate its nominal K-value.
c : Mass (g) of Copovidone in 100 mL of the solution,

calculated on the dried basis.
: Kinetic viscosity of the solution relative to that of water.
pH
Dissolve 1.0 g of Copovidone in 10 ml of water : the pH of
this solution is between 3.0 and 7.0.
Purity (1) Clarity and color of solution - Dissolve 1.0 g of
Copovidone in 10 ml of water : the solution is clear or slightly
opalescent and colorless to pale yellow or pale red.
(2) Aldehydes - Weigh accurately about 1.0 g of Copovidone,
and dissolve in 0.05 mol/L pyrophosphate buffer solution,
pH 9.0 to make exactly 100 mL. Stopper tightly, warm at
60C for 60 minutes, allow to cool to room temperature, and
317
Copovidone
use this solution as the sample solution. Separately, weigh

accurately about 0.1 g of freshly distilled acetaldehyde in a
100 mL volumetric flask containing about 2 mL of water
previously cooled to 4C and add water previously cooled to
4C to make exactly 100 mL. Allow to stand at 4C for about
20 hours, pipet dissolve 0.140 g of acetaldehyde ammonia
trimer trihydrate in water to make 200.0 mL. Dilute 1.0 mL of
this solution, add 0.05 mol/L pyrophosphate buffer solution,
pH 9.0 to make exactly 100 mL, and use this solution as
the standard solution. Measure exactly 0.5 mL each of the
sample solution, the standard solution and water (for blank
test), transfer to separate cells with a path length of 1 cm,
add 2.5 mL of 0.05 mol/L pyrophosphate buffer solution, pH
9.0, and 0.2 mL of -nicotinamide adenine dinucleotide TS
to each of those cells, mix and stopper tightly. Allow to stand
for 2 to 3 minutes at 222C, and perform the test with these
solutions as directed under the Spectrophotometry using
water as the control solution. Determine the absorbances,
At1, As1 and Ab1, of the subsequent solutions of the sample
solution, the standard solution and water (blank) at 340 nm.
Then, add 0.05 mL of aldehyde dehydrogenase TS to each of
the cells, stir and stopper tightly. Allow to stand at 222C
for 5 minutes. Determine the absorbances, At2, As2 and Ab2,
of these solutions in the same manner as above : the content
of aldehyde is not more than 500 ppm (as acetaldehyde).
Content (ppm) of aldehydes as acetaldehyde :
W : Weighed amount (g) of Copovidone, calculated on the

dried basis.
plate, and air-dry the plate. Examine under ultraviolet (main

wavelength : 365 nm) : the Rf value of the fluorescent spot
from the standard solution is about 0.3, and the fluorescence
of the spot from the sample solution corresponding to the
spot from the standard solution is not more intense than
that of the spot from the standard solution (not more than
1 ppm).
(5) Monomers (1-vinyl-2-pyrrolidone and vinyl acetate)
and impurity (2-pyrrolidone) - Weigh accurately 250 mg
of Copovidone into a 10 mL volumetric flask, add 1 mL
of methanol and dissolve by ultrasonic aid. Add water to
make exactly 10 mL, and filtrate this solution if necessary
to remove undissolved particles, and use this solution
as the sample solution. Separately dissolve 50 mg of
1-vinyl-2-pyrrolidone and vinyl acetate each and 300 mg of
2-pyrrolidone in methanol to make exactly 100 mL. Pipet
1 mL of this solution, add a mixture (eluent A) of water,
acetonitorile and methanol (90:5:5) to make exactly 100 mL.
Pipet 5 mL of this solution, add the same mixture (eluent
A) to make exactly 100 mL, and use this solution as the
standard solution. Perform the test with 10 L each of the
sample solution and the standard solution as directed under
the Liquid Chromatography according to the following
conditions, and determine the peak areas, ATa, ATb and ATc,
and ASa, ASb and ASc of 1-vinyl-2-pyrrolidone, vinyl acetate and
2-pyrrolidone in each solution, respectively : the content of
1-vinyl-2-pyrrolidone is not more than 10 ppm, vinyl acetate
is not more than 10 ppm and 2-pyrrolidone is not more than
0.5%. After each test with the sample solution, wash away the
polymeric material of Copovidone from the guard column by
passing the mobile phase through the column backwards for
about 30 minutes at the same flow rate as applied in the test.
Content (ppm) of 1-vinyl-2-pyrrolidone =
W C : Weighed amount (g) of acetaldehyde Concentration

(mg/mL), of acetaldehyde in the reference solution,
calculated from the weight of the acetaldehyde ammonia
trimer trihydrate with the factor 0.72.
Content (ppm) of vinyl acetate =

(3) Peroxides - Weigh exactly an amount of Copovidone,
equivalent to 4.0 g calculated on the dried basis, dissolve
in water to make exactly 100 mL, and use this solution as
the sample solution. To 25 mL of the sample solution add
2 mL of titanium (lll) chloride-sulfuric acid TS, and mix.
Allow to stand for 30 minutes, and perform the test with this Content (%) of 2-pyrrolidone =
solution as directed under the Spectrophotometry, using a
solution prepared by adding 2 mL of 13% sulfuric acid to
25 mL of the sample solution as a blank : the absorbance of
the subsequent solution of the sample solution at 405 nm is
not more than 0.35 (not more than 400 ppm, as hydrogen
W : Weighed amount (g) of Copovidone, calculated on the
peroxide).
dried basis.
(4) Hydrazine - Weigh exactly an amount of Copovidone,
Operating conditions:
equivalent to 2.5 g calculated on the dried basis, transfer
Detector : An ultraviolet absorption photometer (wavelength :
to a 50-mL centrifuge tube, add 25 mL of water, and stir
205 nm and 235 nm)
to dissolve. Add 500 L of a solution of salicylaldehyde in
methanol (1 in 20), stir and warm at 60C for 15 minutes in Column : Stainless steel columns about 4 mm in inside
a water bath. Allow to cool, add 2.0 mL of toluene, stopper diameter and about 25 mm in length, and about 4 mm in
tightly, shake vigorously for 2 minutes, centrifuge, and
inside diameter and about 250 mm in length, packed with
use the upper layer of the mixture as the sample solution.
octylsilanized silica gel for liquid chromatography (5 m in
Separately, dissolve 0.09 g of salicylaldazine in toluene
particle diameter), and use them as a guard column and a
to make exactly 100 mL. Pipet 1 mL of this solution, add
separation column, respectively.
toluene to make exactly 100 mL, and use this solution as the
Column temperature : A constant temperature 30C.
standard solution. Perform the test with these solutions as
directed under the Thin-layer Chromatography. Spot 10 L Mobile phase :
each of the sample solution and the standard solution on a
plate coated with a 0.25 mm layer of dimethylsilanized silica Eluent A : A mixture of water, acetonitrile and methanol
gel with fluorescent indicator for thin-layer chromatography. (90:5:5).
Eluent B : A mixture of water, acetonitrile and methanol
Develop the plate with a mixture of methanol and water
(50:45:5).
(2:1) to distance of about three-fourths of the length of the
318
Copovidone
Gradient :
t [min]
%A
%B
100
100
26
80
20
27
100
36
100
38
100
Flow rate : 1.0 mL/min.

System suitability Selection of column System performance : Perform the
test with 10 L of the standard solution (or appropriate
combination) according to the above operating conditions.
Use a column giving elution of 2-pyrrolidone, vinyl acetate
and 1-vinyl-2-pyrrolidone in this order with resolution
between their peaks being not less than 2.0.
System reproducibility repeatability : When the test is
repeated six times with 10 L of the standard solution
under the above operating conditions, the relative standard
deviations of obtained peak areas of 1-vinyl-2-pyrrolidone,
vinyl acetate and 2-pyrrolidone are not more than 2.0%,
respectively.
(6) Heavy metal - Take 2.0 g of Copovidone, and heat
strongly as directed under the Residue on Ignition. Add 2 ml
of hydrochloric acid to the residue, and proceed according
to JP Method 2, and perform the test. Prepare the control
solution with 2.0 ml of Standard Lead Solution (not more
than 10 ppm).
Loss on drying
Not more than 5.0% (0.5 g, 105C, 3 hours).
Residue on ignition
Not more than 0.10% (1.0 g).
Assay
Vinyl acetate
Weigh accurately about 2 g of Copovidone into a 250 mL
borosilicate glass flask, add an exactly measured 25 mL
of 0.5 mol/L potassium hydroxide-ethanol VS and a few
glass beads, and heat under reflux for 30 min. Titrate
immediately (while still hot) with 0.5 mol/L hydrochloric

acid VS (indicator : 1 mL of phenolphthalein TS) (n1 mL
of 0.5 mol/L hydrochloric acid VS). Carry out a blank test
under the same conditions (n2 mL of 0.5 mol/L hydrochloric
acid VS). Calculate the percentage of copolymerized vinyl
acetate in the Copovidone taken by the formula:
Content (%) of vinyl acetate =
m : Weighed amount (g) of Copovidone, calculated on the

dried basis.
Nitrogen
Weigh 0.1 g in a combustion flask. Add 5 g of a powdered
mixture of 33 g of dipotassium sulfate R, 1 g of copper
sulfate R and 1 g of titanium dioxide R, and wash down any
adhering sample from the neck of the flask with a small
amount of water R. Add 7 mL of sulfuric acid R allowing to
flow down the inside wall of the flask. Heat the flask on an
asbestos wire gauze over a free flame gradually until the
solution has a clear, yellow-green color, and the inside wall of
the flask is free from a carbonized material. Heat for further
45 minutes. After cooling, add cautiously 20 mL of water R,
and connect the flask to the distillation apparatus previously
washed by passing steam through it. To the absorption flask
add 30 mL of a 4 per cent V/V solution of boric acid R,
3 drops of bromocresol green-methyl red solution R and
sufficient water R to immerse the lower end of the condenser
tube. Add 30 mL of strong sodium hydroxide solution R
through the funnel, rinse cautiously the funnel with 10 mL
of water R, immediately close the clamp attached to the
rubber tube, then start the distillation with steam to obtain
80 to 100 mL of the distillate. Remove the absorption flask
from the lower end of the condenser tube, rinsing the end
part with a small quantity of water R, and titrate the distillate
with 0.025 M sulfuric acid until the color of the solution
changes from green through pale grayish blue to pale grayish
red-purple. Perform a blank determination in the same
manner, and make any necessary correction. Calculate the
nitrogen content in the Copovidone taken by the formula:
1 mL of 0.025 M sulfuric acid is equivalent to 0.7004 mg of N
Packaging and storageContainers - Preserve in tight
containers.
319
Contents of the USP Forum
PUBLICATION OF THE LISTS OF CONTENTS

APPEARING IN THE JP FORUM AND USP FORUM
Partners in International Harmonisation
PHARMACOPOEIAL FORUM (USP)
Vol. 32, No. 1
JanFeb 2006
CONTENTS*
STANDARDS DEVELOPMENT
HOW TO USE PF
Section Descriptions
Committee Designations
Staff Directory
POLICIES AND ANNOUNCEMENTS
General Chapters <1> and <905> Postponements
Clarication
USP Issues Notice of Retraction for Residual Solvents
Revisions to Goldenseal Monographs
USP Director of Executive Secretariat Named
Expert Committee Summaries available
on the USP Web Site
USP Announces the Chairs of the Information Expert
Committees
USP Seeks Submission of Proposals for Stability
Indicating Assay Procedures for Steroids
Call for High Priority Monographs for Drug Substances
and Products, and Excipients
Pharmacopeial Education Courses
Visit the USP Web Site at http://www.usp.org
International Correspondence
How to Submit Comments
New Pharmacopeial Forum Public Review and Comment
Period Deadlines
Publication and Comment Schedule
Publication Schedules
FIRST INTERIM REVISION ANNOUNCEMENT
USP MONOGRAPHS
Lithium Carbonate Extended-Release Tablets
DIETARY SUPPLEMENTS - MONOGRAPHS
Goldenseal
Powdered Goldenseal
Powdered Goldenseal Extract
ERRATA LIST FOR USP 29NF 24
IN-PROCESS REVISION
USP MONOGRAPHS
Acetazolamide Oral Solution [new] (USP 30)
Acetazolamide Oral Suspension [new] (USP 30)

Albendazole Oral Suspension (USP 30)
Alprazolam Oral Suspension [new] (USP
30)
rd
Amoxicillin Capsules (Proposal for 3 IRA)
Azathioprine Oral Suspension [new] (USP 30)
Baclofen Oral Solution [new] (USP 30)
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Benazepril Hydrochloride Tablets [new] (USP 30)
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rd
Clopidogrel Tablets (USP 30) (Proposal for 3 IRA)
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(USP 30)
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(USP 30)
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Dronabinol (USP 30)
Felodipine
Extended-Release Tablets (Proposal
rd
for 3 IRA)
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Irbesartan (USP 30)
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(USP 30)
* The USPNF (USP 30NF 25), the Supplement (Supp), or the Interim Revision Announcement (IRA) for which the revision proposal is targeted
is shown in parentheses next to each proposed item.
320
Contents of the USP Forum
Labetalol Hydrochloride Oral Suspension [new] (USP 30)

Lovastatin (USP 30)
Mebendazole Oral Suspension (USP 30)
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Ginkgo Tablets [new] (USP 30)
NF MONOGRAPHS
Acetyltributyl Citrate (NF 25)
Acetyltriethyl Citrate (NF 25)
Cellacefate (NF 25)
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Tributyl Citrate (NF 25)
Triethyl Citrate (NF 25)
GENERAL TEST CHAPTERS
<11> USP Reference Standards (USP 30)
<231> Heavy Metals (USP 30)
GENERAL INFORMATION CHAPTERS
<2040> Disintegration and Dissolution of Dietary
i
Supplements
(USP 30)
REAGENTS, INDICATORS AND SOLUTIONS
Reagent Specifications
Dextran, High Molecular Weight (USP 30)
Hydrazine Hydrate, 85 % in Water (USP 30)
1-Naphthol (USP 30)
p-Toluenesulfonyl-L-arginine Methyl Ester Hydrochloride
(USP 30)
REFERENCE TABLES
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HARMONIZATION
DIETARY SUPPLEMENTS - MONOGRAPHS
PHARMACOPEIAL PREVIEWS
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STIMULI TO THE REVISION PROCESS

Instructions to Authors
NOMENCLATURE
INDEX
_______________________________________________________________________________
CORRESPONDENCE
Individuals who wish to correspond with the JP or the USP concerning any of the monographs/articles mentioned
can do so at the following addresses:
Japanese Pharmacopoeial Forum
Pharmacopeial Forum
Secretariat of the Japanese Pharmacopoeia

Evaluation and Licensing Division
Pharmaceutical and Medical Safety Bureau
Ministry of Health and Welfare
1-2-2, Kasumigaseki, Chiyoda-ku
Tokyo, 100-8045, JAPAN
The United States Pharmacopeia

Division of Standards Development, USP-NF
12601 Twinbrook Parkway
Rockville, Maryland 20852
USA
321
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322

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PHARMEUROPA 18.

New: Pharmeuropa, Pharmeuropa Bio and

Draft Monographs and General Texts

Publication of Supplements 5.6 and 5.7

180 Avian infectious bursal disease vaccine (inactivated)

Annual Report of Activities of the EDQM

181 Cholera vaccine (inactivated, oral)

Electronic version of the 5th Edition of the

Drop point (2.2.17)

CRS: conditions of sale

List of CRS adopted at the November 2005 session

List of texts adopted at the November 2005 session

Updated work programme of the European Pharmacopoeia

Herbal drugs: sampling and sample preparation (2.8.20)

Lincomycin hydrochloride monohydrate

Macrogol 40 sorbitol heptaoleate

List of codes of groups of experts

List of Standard Terms: 5th Edition (2004)

Standard terms: new and revised terms

Elaboration / Revision of a monograph (Procedure 1)

Technical Guide: 4th Edition (2005) available

Proceedings of conferences of the EDQM

Pharmeuropa Scientific Notes

Pantoprazole sodium sesquihydrate

Preparations for nebulisation: characterisation (2.9.44)

Methylergometrine hydrogen maleate

213 Pyridoxine hydrochloride

Training Sessions on the 5th Edition of the European

Rabbit haemorrhagic disease vaccine (inactivated)

27-28 April 2006, Chicago, USA

13-14 November 2006, Dublin, Ireland

St. Johns wort dry extract, quantied

Streptokinase concentrated solution

Impurities Control: Setting Specifications

New Microbiology Chapters of the European

Vaccines for veterinary use

Illustrations of powdered drugs in herbal monographs:

Certification of Suitability of the

231 Eucalyptus leaf

Herbal Reference Standards

PHARMEUROPA Vol. 18, No. 2, April 2006

THE EUROPEAN PHARMACOPOEIA

PHARMEUROPA Vol. 18, No. 2, April 2006

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PHARMEUROPA Vol. 18, No. 2, April 2006

The EDQM HELPDESK

IF YOU HAVE A QUESTION

WHAT ARE THE ADVANTAGES OF USING

HOW CAN I CONTACT AND/OR SEND

For more information, visit:

PHARMEUROPA Vol. 18, No. 2, April 2006

Annual report of activities of the EDQM

European Directorate for the

the European Pharmacopoeia, including

the procedure for Certication of Suitability of

the European network of Ofcial Medicines

In 2005, pharmacopoeia authorities and licensing

control laboratories continued to yield encouraging

Annual report of activities of the EDQM

Summary of agreements on harmonisation by the PDG

and uniformity of dosage units are in preparation for

PHARMEUROPA Vol. 18, No. 2, April 2006

Annual report of activities of the EDQM

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