LOGFILE No.
1 / January 2013
Good practices for Pharmaceu-
tical Microbiology Laboratories
Author: Tim Sandle
This article is republished with permission
from GMP REVIEW, VOL.11 NO.3
OCTOBER 2012.
A review of the WHO Guidance
Introduction
In late 2011 the World Health Organisa-
tion (WHO) reissued Annexe 2 of its
Technical Report 961 (Specifications
for Pharmaceutical Preparations). The
Annexe consists of a guideline entitled
Good Practices for Pharmaceutical
Microbiology Laboratories. The guid-
ance for pharmaceutical microbiology
laboratories was originally written in
2009.
The reason for the relatively fast revision
came about due to concerns from WHO
GMP inspectors who reported that aspects
of the guideline appeared either open to
interpretation or had proven difficult for
some laboratories to understand.
The Good Practices document has some
similarities with the USP chapter <1117>
Microbiological Best Laboratory Practic-
es and together the two chapters are of
importance given that there only other
significant regulatory document is the FDA
inspection guide Microbiological Pharma-
ceutical Quality Control Labs, which is
now a little out of date and was last re-
vised by the FDA in 1993. With regard to
European GMPs and compendia there is
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no equivalent guidance or pharmacopoeial
chapter.
Analysis
The WHO document has a relatively nar-
row scope in terms of the perceived activi-
ties of a pharmaceutical microbiology
laboratory and the scope is certainly in-
wards, in terms of the way a laboratory is
run, rather than focusing outwards on the
activities which laboratory staff might
undertake. For example, the document is
concerned with viable microbiological
environmental monitoring within the labor-
atory but not with the activities of microbi-
ology staff going into process areas to
take airborne particle counts and to collect
viable monitoring samples. This seems a
little limited since the case for monitoring
in an unclassified laboratory is somewhat
debatable and the latter is something
which occupies the time of many laborato-
ry staff.
The guide has a glossary at the front in
which a relatively short number of terms
are defined. Here there are definitions of
precision and robustness but not of
environmental monitoring or bioburden. I
am not altogether certain how useful this
list is and the scope of the definitions is
either vague or lacks sufficient detail to be
meaningful. Take, for example, reference
cultures: Collective term for reference
strain and reference stocks
Either the reader knows what is meant by
a reference culture and its purpose for
quality control and in which case the defi-
nition is superfluous; or the reader does
not know and in which case the definition
is not very informative.
The first main section of the document
discusses personnel: staff who should be
working in the microbiology laboratory.
Here the requirement is limited. The doc-
ument states that testing should be carried
out and supervised by staff with experi-
ence and who hold an unspecified qualifi-
cation. Whilst this is important the docu-
ment disappointingly does not state that
Page 1/4
LOGFILE No. 1 / January 2013
staff should hold a degree in an applicable
bioscience subject.
The document is useful, however, in stat-
ing the need for supervision and for au-
thorised personnel for the release of re-
sults from the laboratory. The document
also outlines some of the basic training
required for staff including aseptic tech-
nique, serial dilutions, working with haz-
ardous cultures, and colony counting.
Although there is not a great deal of detail,
the section is important because common
agreement, nationally or internationally, on
microbiology laboratory training is lacking.
The second substantive section concerns
laboratory premises. In this part reference
is made to the need for dedicated areas
for laboratory equipment, layout for opera-
tions, the separation of laboratories from
production areas, and controlled access to
microbiology areas. Reference is also
made to cleaning and disinfection of the
laboratory, spillage policy and hand saniti-
sation.
There is one statement in this section
about Sterility Testing which makes an
incorrect assumption that isolators are not
being widely used regarding the classifica-
tion of the room in which sterility testing
takes place. With the sterility test environ-
ment a note is made about clean air clas-
sification being annual. This is out of step
with ISO 14644 which requires the ISO
class 5 clean air device to be classified six
monthly.
Reference to environmental monitoring of
sterility test areas and the trending of
results is made.
The third main section concerns the vali-
dation to test methods. This is a relatively
short section compared with USP <1227>.
The section sates that pharmacopoeial
test methods are considered validated
but that specific products, when used
against compendial test methods, must be
shown to be suitable in terms of recover-
ing microorganisms. There is a reference
to the use of alternative test methods with
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a generalised list of validation require-
ments.
The fourth section concerns equipment, in
terms of maintenance and calibration. The
guidance is for a documented programme,
which is useful as some laboratories do
not have a calibration procedure or clearly
defined acceptance criteria. References
are made to national calibration standards
and the need to assess items like pipettors
and laboratory timers.
There is also some useful guidance for
setting calibration intervals by tracking the
period of drift for instrumentation from
normal operating conditions. Special em-
phasis is given to temperature monitoring
devices and incubators, which are critical
for culture based methods. There are also
sections on autoclaves and balances. With
the former, on the oft debated issue of
whether biological indicators should be
used for laboratory autoclaves, there is no
mention.
The fifth section concerns reagents and
culture media. With reagents, there is a
reference for the need to assess shelf life.
The part on culture media is longer and
there is mention of both media prepared
in-house and media purchased from a
vendor. With regard to growth promotion,
in keeping with most laboratories, the
requirement is to test each batch. There is
also a statement about testing each ship-
ment, which some might regard as exces-
sive. There is a further stipulation about
assessing transport requirements. This
latter point is something which is not al-
ways addressed.
The test requirements for culture media
and the acceptance criteria are the same
as the harmonised microbial enumeration
chapters (USP <61> and Ph. Eur. 2.6.12).
The preparation of media, in keeping with
cGMP, is addressed as per any production
batch manufacture.
The media section also includes a part on
the resuscitation of microorganisms, which
is quite useful.
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LOGFILE No. 1 / January 2013 Maas & Peither AG GMP Publishing

The sixth section addresses reference

materials and cultures. The advice for Topic WHO USP FDA
storing cultures, subculturing and limiting Alert and action levels Yes No No
the number of passages is consistent with Aseptic techniques No In No
the pharmacopoeias. part
Autoclaves Yes No No
The seventh and eighth sections overlap a Balances Yes No No
little and address sampling and sample Cleaning and Yes No No
handling. With sampling, the need to test disinfection
samples promptly and to keep chilled Contaminated waste Yes No No
where appropriate is useful but there is disposal
little with regard to sampling methods and Contract test facilities No No Yes
no mention of aseptic techniques. The part Culture media: manu- Yes Yes Yes
facturing
relating to sample handling describes
Culture media: testing Yes Yes No
rudimentary logging procedures.
Deviations/OOS No No No
Environmental moni- Yes No No
The ninth section is very short and em-
toring of micriobiolgy
phasises the importance of contaminated laboratory
waste disposal. The tenth part is also brief Equipment calibration Yes In No
and describes the importance of internal part
laboratory quality control (laboratory con- Equipment Yes In No
trols). This section really should contain qualification part
more information and a description of Laboratory de- Yes Yes
laboratory deviations is clearly lacking. sign/premises
Laboratory manage- In Yes Yes
The eleventh section is about testing ment part
procedures and does not go into any Laboratory staff Yes Yes No
meaningful detail. The twelfth and final Method validation: Yes No No
section addresses test reports and touch- alternative methods
es on the debate on reporting quantitative Method validation: Yes In Yes
results: is it not detected for a defined pharmacopoeial part
unit or zero? Microoraganisms: use Yes Yes No
in the laboratory
The WHO document is supported by some Microorganisms: Yes Yes No
appendices on classifying different parts of reference cultures
the laboratory environment and some Non-sterile product No No Yes
tests
suggested maintenance periods for vari-
Quality control Yes No No
ous items of equipment, which seems a
Reagents Yes No No
little out of place to me as this would de-
Reference cultures Yes Yes No
pend very much on the item, the manufac-
Sampling Yes Yes No
turer and the performance of the equip-
Sterility testing envi- Yes No Yes
ment in the laboratory.
ronment (cleanroom)
Temperature Yes In No
Comparison of WHO, USP and FDA monitoring devices part
Test procedures Yes Yes No
The WHO document, despite some limita- Test records Yes Yes Yes
tions, does stand as a useful adjunct to the Training In Yes No
USP chapter and the FDA inspection part
guide. To give an overview, I have high- Waste disposal Yes In No
lighted similarities and differences in the part
table below.

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LOGFILE No. 1 / January 2013
Summary
The WHO document is a mixed affair. In
some areas there is no mention of new
technologies or the operation of common-
place technology like Laboratory Infor-
mation Management Systems (LIMS); in
other areas it lacks any detail (such as
reagents), in others it provides unhelpful
information (such as the parameters for
alternative methods), yet in other cases it
provides useful advice, especially in terms
of laboratory equipment, or at least
thought provoking material, as with the
test requirements for culture media. It also
shies away from, fortunately in my view,
any references to saving time, money or
resources. A quality based guide should
focus on just that best practice and
leave the budgetary aspects to other types
of publications.
This assessment of the WHO guidance
has been generally critical and this reflects
the fragmented and inadequate nature of
regulatory guidance on microbiology la-
boratories, where there is a paucity of
regulatory guidance within GMPs and
pharmacopoeias. The guidance which is
available is lacking information or does not
go into sufficient detail in order for the
laboratory manager to rely on such docu-
ments alone in establishing best practices.
Furthermore, the information presented
within different guidances is often contra-
dictory.
There is a clear void to be filled with the
need for a clear, concise and practical
guide to the operation of microbiology
laboratories operating in a GMP environ-
ment.
References
Food and Drug Administration. Guide to
Inspections of Microbiological Pharmaceu-
tical Quality Control Laboratories (July
1993). At:
www.fda.gov/ICECI/Inspections/Inspection
Guides/ucm074914.htm
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USP. 2010. <1117> Microbiological Best
Laboratory Practices.
USP 33
World Health Organisation. Good Practic-
es for Pharmaceutical Microbiology Labor-
atories. At:
http://apps.who.int/prequal/info_general/do
cuments/TRS961/TRS961_Annex2.pdf
About the Author:
Tim Sandle is Head of Microbiology at
BioProducts Laboratory. He is currrently
chairman of the Pharmig LAL Action
Group and serves on the UK Blood Ser-
vice Cleaning and Disinfection Committee.
He has written and contributed to many
papers and books, including "Microbiology
and Sterility Assurance in Pharmaceuticals
and Medical Devices" and "Cleanroom
Management in Pharmaceuticals and
Healthcare".
E-Mail: tim.sandle@bpl.co.uk
Web: www.pharmig.blogspot.com
This article is republished with permission
from GMP REVIEW, VOL.11 NO.3
OCTOBER 2012.
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