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The lab report should use the template in the file “Report form – Rami.doc”.
The methods used to analyse these samples are described in the file “Methods used to obtain the data – Rami.doc”.
The data obtained is described in the file “Bioburdens data – Rami.doc”
You will need to provide a title, an abstract (max 125 words), results (max 125 words, excluding tables and calculations), discussion (max 1100 words), conclusion (max 100 words), and references (up to 10 references).
You will not need to design your own introduction. However, you will need to select the best introduction for your lab report from the options (A, B, or C) offered in the file “Available introductions – Rami.doc”. You will need to explain the reasoning of your choice (max 50 words).
You will not need to design any material and methods section.
No annexes are requested for this lab report.
MICROBIOLOGICAL EXAMINATION OF PHARMACEUTICAL
Safety considerations and precautions
This practical involves the use of live cultures which must be considered as potentially
pathogenic and therefore handled appropriately. However, it is not necessary to wear gloves
unless cuts, grazes, skin rashes or open sores are present on the hands. Please ensure that
you are familiar with the regulations concerning safe handling of microorganisms which are
given in the front of your laboratory booklets (SAFETY INFORMATION, Section 1: Safety in
Microbiology Labs). All items of lab ware must be disposed of correctly. Please refer to the
information given in the front of your booklet (SAFETY INFORMATION, Section 2: Safe
disposal of used lab ware & contaminated cultures) or ask a member of staff if you are
unsure of how to dispose of your items. NEVER place contaminated items down on the
bench or other surfaces. Please ensure you know how to proceed in the case of a spillage
(SAFETY INFORMATION, Section 3: What to do in the event of a contaminated spillage). A
COSHH and a Risk assessment for this lab class are available in the annex of this laboratory
schedule (page 50). Please read these documents before the laboratory class and pay
particular attention to the need of using gloves when disinfecting the benches, and to the
risks of using Bunsen burners.
The microbial quality of pharmaceutical raw materials and manufactured medicines is
determined in two principal ways: by placing limits on the number or concentration of
microorganisms that may be present (bioburden) and by excluding certain objectionable
organisms altogether. The bioburden of raw materials is a relevant parameter when
designing sterilisation processes. The level of sterility assurance achieved is dependent
upon the design of the sterilisation process itself and upon the pre-sterilisation bioburden of
those materials. The determination of bioburdens uses standard microbiological methods
such as pour and spread plates, and membrane filtration, but the application of these
methods to raw materials and manufactured medicines is less straightforward than when
performing counts on simple aqueous suspensions of bacteria.
The purpose of this work is to illustrate how different characteristics of materials (e.g.
formulation, viscosity) and the absence of any indication of the approximate level of
contamination have to be considered when conducting bioburden determinations.
Preparatory reading and visual materials
You will be asked to select a protocol to determine the bioburden of three different
pharmaceutical products. To aid you in this task we strongly advise you to carry out some
preparatory reading prior to the lab. Use the references below to attain information on the
• Types and concentrations of microorganisms which are commonly found as
contaminants of pharmaceutical raw materials and manufactured medicines. What
factors influence the types and concentrations?
• Principal methods available for determining the levels of microbial contamination in
pharmaceutical products. Are there any types of formulation for which these methods
might be unsuitable?
• What special measures would you need to take for estimating the bioburden of a
product that is viscous, oily or a suspension?
• What issues arise from the inclusion of preservatives in some formulations? How
would you deal with these when estimating microbial bioburdens?
1. Hanlon, G. and Hodges, N. (2013). Essential Microbiology for Pharmacy and
Pharmaceutical Sciences, Wiley-Blackwell. ISBN9780470665343: Chapter 15
(Bioburdens: counting, detecting and identifying microorganisms)
2. Baird, R. M., Hodges, N. A. and Denyer, S. P. (2000) Handbook of Microbiological
Quality Control: Pharmaceuticals and Medical Devices, Taylor and Francis. ISBN
074840614 X.: Chapter 4 (Enumeration of microorganisms)
3. Denyer, S. P. and Baird, R. M. (2007) Guide to Microbiological Control in
Pharmaceuticals and Medical Devices, 2nd Edition, CRC Press. ISBN 0748406158:
Chapter 8 (Monitoring microbiological quality: conventional testing methods)
4. Aulton, M.E and Taylor, K.M.G (Eds). (2013) Aulton’s Pharmaceutics: The design
and manufacture of medicines, 4th Edition, Churchill Livingstone
ISBN9780702042904: Chapter 14 (Pharmaceutical applications of
You will need to perform several microbiology methods to implement your protocols. All
these methods were formerly addressed at the conjunctivitis case. To revise how these
methods are executed, we strongly advise you to watch the videos below prior to the lab.
How to use a pipette and pump How to use a micropipette
How to perform serial dilutions (I) How to perform serial dilutions (II)
How to prepare pour plates How to prepare spread plates (I)
How to prepare spread plates (II) How to carry out membrane filtration
Work in pairs.
Partner’s name (s)
You are provided with the following samples of three pharmaceutical products:
1 An aqueous solution (100 ml) suspected of containing a concentration of aerobic
bacteria estimated to be between 1 and 104 CFU/ml, labelled contaminated aqueous
2 A lotion (preserved with parabens) suspected of containing a concentration of aerobic
bacteria estimated to be between 102 and 106 CFU/g, labelled contaminated lotion.
3 A sample of talcum powder suspected of containing a concentration of aerobic bacteria
estimated to be between 102 and 106 CFU/g, labelled contaminated talc.
Note: the aqueous solution and the lotion are examples of non-sterile pharmaceutical products; talc is
an example of a raw material.
You are provided with the following materials:
30 Nutrient agar plates (over dried)
2 u 90 ml sterile water
Sterile water (unmeasured volume)
50 ml sterile Lecithin-Tween NB
1 sterile membrane filter unit
Membrane filters, 0.45 Pm (3u)
Sterile Pasteur glass pipettes and respective teats
Sterile boiling tubes or sterile universals (15u)
Micropipettes (P200, P1000) and respective tips
10 ml sterile pipettes and respective bulbs
Two spatulas (non-sterile)
Before you begin: Your partner and you will need to agree about the most appropriate
protocol to be implemented from the options (1, 2, 3, 4, 5 or 6) offered in the annex of the
laboratory schedule (pages 37-49). You will be permitted 30 min maximum to discuss with
your partner about the best protocol to use and why.
From the six protocols (1, 2, 3, 4, 5 or 6) offered in the annex of the laboratory schedule,
which one will you and your partner use in the laboratory to analyse the contaminated
Select option 1, 2, 3, 4, 5 or 6
Implementation: Using the materials provided, and the selected protocol, perform viable
counts on each of the samples given to you to calculate the bioburden. Your experimental
work will be based on the equipment/materials available as you will not be provided with
additional materials. The lab bench should be swabbed down with disinfectant both before
and after work has been carried out (wear gloves during this procedure).
There will be an opportunity to read your results at the next laboratory session and the lab
report can be written when these results are available.
Note: you must remain in the laboratory for both the planning and practical stages of the
RESULTS AND REPORT
You can use the following boxes to record your results for each of the three products
examined (draw up a table of results from your own data).
Table of results: Contamination of an aqueous solution
Table of results: Contamination of a lotion
Table of results: Contamination of talcum powder
1. Each student should prepare an individual lab report, using the Word template
available from StudentCentral, under the Quality of Sterile Medicines case (folder
‘Study Materials and Reading List’);
2. Please refer to Coursework Assessment (pages 7-8 of this Lab Schedule) for more
information about the lab report, including assessment criteria, submission and