The document discusses auditing of microbiology laboratories. It provides definitions of auditing and outlines areas that should be assessed such as laboratory layout, equipment and facilities, documentation practices, and manufacturing processes. Key areas that are important for auditors to evaluate include laboratory organization, sampling procedures, media preparation, equipment maintenance, method validation, documentation, biosafety, and proficiency testing. The role of the microbiology laboratory in auditing sterile product facilities is also described.

1. AUDITING OF MICROBIOLOGY
LABORATORY
BY: NILAM ASHOK GHADAGEPATIL
FIRST YEAR M. PHARM PQA (SEM-II)
GUIDED BY : PROF. AMOL. S. BANSODE
DATE:13 JUNE 2022

2. Index
 Definition
 Layout
 Auditing microbiology laboratory
 General documentation
 Role of the microbiology laboratory in auditing sterile products manufacturing
facilities
 Auditing and manufacturing processes.
 General area of interest
 Key references
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3. Definition: Audit
 “A service involving an independent review of an organization’s records,
operation and procedure to evaluate for efficiency, effectiveness, compliance
and the existence of adequate internal controls to mitigate risks to the
achievement of the organization’s objectives.”
 Objective:
 To ensure quality of the product.
 To assess effectiveness of QA system.
 It permits timely corrections of problems.
 It established high degree of confidence
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4. layout
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6. Auditing microbiology laboratory
 Good house keeping:
• Organization and cleanliness of the laboratory.
• Cleaning and disinfection process.
• Data of expiry dates agents.
• Followed GMP
• House keeping practices should be in English as well as
local language
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7.  Sampling and sample receipt:
 Storage
 Traceability
 Batch record
 Good practices
For this review ,appropriate questions to ask may include:
• How samples are taken in production?
• What protection afforded?
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8.  Does the microbiology team provides sampling training to
operators to ensure sample are taken aseptically.
 Are sample stored until end of date or stored in refrigerator or
passed further?
 Have these sampling procedure been appropriately assessed?
 Does log book provided for who delivered sample and then
who took it for testing?
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9. Culture media
 It is important for the auditor to assess the laboratory is using
the correct media. This include assessing the components in
making media based on the use of accepted sources or
references for formula, whether the medium is manufactured
in house or purchased from an external supplier.
 If media is prepared in house, some appropriate audit points
to consider and questions to ask are:
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10. • Review the SOPs for media preparation.
• Assess the calibration and daily verification of balances.
• Check the suitability of glassware and washing procedures.
• Assess the sterilization process
• Labelling
• Storage temperature are appropriate
• Is media stored away from sunlight.
• To assess that storage away from sunlight.
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11. Microbiology laboratory equipment:
 Microbiology laboratory equipment needs to be well maintained.
 New equipment need to be qualified before use.
 URS, DQ, IQ,OQ, PQ
 Use should be recorded in log books
 To support the operation of equipment, many items require on-going
checks to be performed at frequencies.
 Incubators, refrigerator and freezers should be monitored and should
operate within their defined limits.
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12.  Where autoclaves are used, it is important to assess that autoclave cycles
are monitored and records kept of loads processed, test like:
• leak test-not more then 1.3 bar per min.
• empty chamber test heat distribution study
• container mapping
• steam penetration study
• microbial challenge study are performed
 It is also important that equipment is frequently cleaned and disinfected
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13. Method validation and qualification:
Samples assayed against pharmacopeial(official test) methods have been
appropriately verified and that any non-pharmacopeial(non official test)
methods have been verified .
Sample qualification can include:-
 Bacterial endotoxin test:
Product can be contaminated during purification, production or packaging
stages, test is used to confirm that the product are not contaminated.
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14.  Water testing
 Water samples are tested
 Purified water used should be as per WHO norms
 Sterility testing:
 Auditor assess the technique for samples media test equipment and environment
 Antibiotic assay:
 Audit should consider the control and preparation of standard as well as sample
API used.
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15. General documentation:
• The auditor will review the documentation in the microbiology laboratory
from SOPs to results sheets and computerized system.
• Auditor will seek assurance that correct and up to date procedure are
begin used and that testing histories are accurate and complete.
Some examples for auditor to check for on documentation include:
• Date of test
• Material tested
• Batch no. lot no. of material
• Microbiologist names
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16. • Procedure number and version
• Reagent used and expiry date
• Equipment used
• Incubation conditions
• Date of test reading
• Documented test result
• Acceptance criteria
• Supervisor or management sign off
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18. Biosafety:
The microbiological laboratory poses risks to laboratory staff and to
visitor.
 In terms of risks posed by specific microorganism, the World Health
Organization guidance is:-
Risk group1: Agents not associated with disease in health adult humans.
Risk group 2:Agents associated with human disease that is rarely serious.
Risk group 3: Agents associated with serious or lethal human disease.
Risk group 4: A pathogen that usually causes serious human or animal
disease
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19.  Protective measure:
Use of biosafety cabinets
Use of personal protective equipment.
Good waste disposal practices.
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20. Proficiency testing
• Self –audit can be done by joining proficiency testing scheme
• Determines the performance of individual laboratories.
• Proficiency testing is also called interlaboratory comparison.
• The result reported by each laboratory for measured are compared to
reference value.
• Helps to assess the suitability of test method and the appropriateness of
laboratory workflow
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21. Role of the microbiology laboratory in auditing
sterile products manufacturing facilities
• Microbiology laboratories have certain equipment (e.g. autoclave,
glassware) should be dedicated and separate from other areas.
• Designed to suit the operation to be carried out in them.
• Should have adequate space.
• Take in account construction material.
• Separate air supply, air handling unit.
• Personnel
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22.  Sample preparation should be segregated.
 Laboratories equipment should not be used outside
 HEPA filters
 Air flow pattern.
 Changing room
 Microbial environment monitoring of sterility test zone should be
performed during every work session.
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23. Auditing and manufacturing processes
FACILITIES:
• Clean areas should be clearly separate from toilet, cafeterias.
• Should be suitable size.
• Individual area should be separate.
• Ceiling should be effectively air-sealed.
• Proper designing of doors and windows
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24. • Sealing should be sealed to prevent contamination.
• Operation of filling& sealing should be controlled, supervised and recorded.
• Sanitary pipes & fitting should be used and threaded pipe connection should
be avoided.
• Air lock should not be opened simultaneously.
• Filtered air supply should be used to maintain positive pressure.
• Warning system should be operated to indicate a failure in air supply.
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Facilities Instruments

26. A) EQUIPMENT:
Equipment used in manufacturing of sterile product may include:
• Production equipment
• Container/closure
• Support system/material system related equipment.
 Aseptic Processing Equipment:
• Determine that all equipment that comes in direct contact with the product are
sterilized
• Equipment that come in direct of product should be sterilized
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27.  Stopper washer
• Quality of water used for washing and associated water sampling /qualification
data.
• The appropriateness of the air supply used in drying operations should also be
verified.
 Capping Equipment (vials):
• Vial cap provides the final closure element of a sealed vial.
• Cap on vial protects the stopper from external damage.
• Evaluate preventive maintenance schedules of the capping machine.
• Air supply quality to the capping unit should also be evaluated
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28. Sterilizers
• Inspection should cover IQ,OQ and PQ.
• The inspection of sterilizers should include a physical examination of
equipment.
• Engineering specification which may described in equipment's DQ
• Sterilizer is maintained calibrated and drained properly. (DQ)
• Record of unplanned maintenance as well as preventive maintenance,
should be reviewed
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29.  Lyophilizer:
• Partially filled vials are used in lyophilisation process, sterile product is
exposed to environment from time of filling until fully sealed.
• The inspection should verify that partially sealed vials are transported & in
Lyophilizer under class 100(ISO 5) protection.
• Validation of sterilization of lyophilisation chamber between uses, current
sterilization controls, leak testing of the chamber, integrity testing of air/
gas filters, and calibration of temp and pressure controllers.
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30.  Restricted access barrier system (RABS):
 RABS provides a physical barrier between workers and production area.
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31. Two types of RABS
1. Closed RABS
2. Opened RABS
 When inspecting RABS
• Determine that the gloves and gauntlets attached to the glove ports are sterile when
installed.
• Well defined procedure that describe what is done when an open door intervention is
performed
• Checking of products coming in contact
• Verify that all process under gone through disinfection.
• RABS entry is often accompanied by an appropriate line clearance, which should be
documented in batch record
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32.  Blow-fill-seal(BSF) technology
 BSF is an automated aseptic filling process in which containers are formed, filled and
sealed in continuous operation.
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33. When inspecting BFS:
• Verify that HEPA-filtered or sterile air is used .
• Evaluate preventive maintenance program.
• Determine the sterilization cycle has been validated and condensate
properly drains from line.
• Verify personnel.
• Observe equipment setup.
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34.  Sterile filtration:
• The objective of aseptic processing is to maintain the sterility of
product.
 Operating condition
• The environment
• Personnel
• Container/closure
• Sterilizing filter
• Maximum holding period of product
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35. General area of interest:
A)Buildings:
• Minimum standard to prevent the infestation of birds and rodent into manufacturing
area.
• Unnecessary entry should be avoided.
• Grade A and B areas should be designed so that operation can be observed from
outside.
• Exposed surface should be smooth, unbroken, min shedding.
• Cupboards Pipes and ducts
• Grade A,B,C. Doors
• Warning system
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36. B) RAW MATERIAL
 Sop’s should be established for acceptance, inspection, storage, sampling
and testing of raw material.
 Sterility of raw material is recommended to be ensured by periodic
monitoring.
 If raw material depyrogenated during the manufacturing process, suitable
depyrogenation procedure should in instituted
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37. C) WATER
Purified water:
 Purified water is water that has been mechanically filtered.
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38. A)Distillation :
 Distillation involves boiling of water and then condensing the vapour into a clean
container, leaving solid contaminants behind.
 Distilled water is produced by distillation.
B)Demineralization:
 Demineralization is removing all the material that can be found in natural water.
 This is done when water is used for chemical process.
 All chemist and beauty product are made by demineralized water
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39. D) Packaging materials:
Ampoules and vials:
 Containers for parentral preparation are made from uncoloured glass
except that coloured glass may be used for substances known to be
light sensitive.
Evaluation test:
I) Powder glass test: It is done to test amount of alkali leached from
powder glass
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40. Powder glass test
10gmsample is added with 50 ml of high purity water
in 250 ml flask
Place it in autoclave at 1210c for 30 min
Cool it under running water
Transfer to another flask& wash again with 15
ml high purity water.
Titrate immediately with 0.02 sulphuric acid, using
methyl red as indicator and record volume
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41.  Key references
 Prof. Manohar A. Potdar, Simran Shaikh, Dr. Prem R. Goel. Text Book Of Audit and
regulatory compliance, pp.4.1-4.52
 Rogers, Bywater, M.J., and Reeves, D.S(1991) Audit of turn-around times in a
microbiology laboratory, J Clin Pathol. 44(3):257-258
 Mifsud, A.J and Shafi, M.S. (1995) Internal audit in a microbiology laboratory, J Clin
Pathol.48(6):560-563
 Sandle.T. (2015) Approaching Microbiology Method Validation, Journal of GXP
Compliance,19(4):1-15
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42.  Albertini. A. and Signorini .C.(1995).The Quality Assurance system in clinical
chemistry.31(1):3-8
 Farrington M .(1992) Medical Audit in clinical microbiology in united
kingdom.Rev Med Microbial 3:104-11
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