Microbiology

Laboratory Audit

A Check List for the Pharmaceutical Industry

Archana Pandit B. Sc. Microbiology
GMP Scientific Inc.
Newtown, PA 18940
 Foreword

No checklist, however comprehensive, can replace a well directed audit conducted by a

competent auditor. This one is no exception.

Over the years, I had wanted to have a list of items I use to review while conducting a
Microbiology laboratory audit. I never used such a check list per se, but I used a hand
written list of items that I considered to be most important. This list got longer or shorter
depending on the type of laboratory I was auditing, and the activities performed in that
lab. This, of course took time, and I ended up writing lot of similar things for most
laboratories. Finally, I decided to create the one that I wanted, partly to avoid making
those hand written lists and partly to save time.

I have provided some pointers about general types of things applicable to a laboratory in
a pharmaceutical operation. I have also used information from the FDA Guidance on
Microbiology Laboratory audit to make it more complete. This checklist does not tell you
how to do the audit and what standards you should use to make your observations. I leave
that to you.

Eventually, I will supplement this checklist with a more detailed guidance. There are
several books out there on how to conduct an audit and I am sure your company has
auditing procedures.

With that, I want to remind you that this is by no means an exhaustive, all inclusive list.
No warranties are made about its completeness, accuracy, and appropriateness and no
liabilities are assumed, if you use this check list. It is just an aid. But I do hope it saves
you time and money.

Feel free to write to me at gmpscientific@gmail.com, if you have any suggestions, or

comments. I will look forward to make it better.

Archana Pandit
GMP Scientific Inc.
Newtown, PA 18940

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 MICROBIOLOGY LABORATORY AUDIT CHECK LIST*

ORGANIZATION/PERSONNEL/MANAGEMENT

Review the organization and reporting structure for laboratory personnel.

Review

- Organizational Structure
- Sample job descriptions
- Qualification for a representative number of employees (CV, Certifications)
- Training procedures (GMP and job specific)
- Training records (Planned and for cause)
- Investigation follow-ups where employee training was found to be inadequate
- Adequate personnel versus work load
- Adequate supervision

Observe employee practices in the laboratory, specifically where aseptic techniques

are employed.

Evaluate whether, adequate employees and supervision is provided through out the
laboratory operations.

Evaluate how Management is notified of critical laboratory issues and the level of
management involvement.

Review whether laboratory employees are included in the environmental monitoring

program (e.g. using surface contact plates).

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GENERAL LABORATORY

Review laboratory space, design, layout, access, workflow, and organization for
suitability of its purpose.

Review the cleaning and sanitization schedules, environmental monitoring programs

(viable and non-viable where applicable), respective procedures, and sample records
of such data.

Verify the effectiveness of the cleaning and sanitization schedules and its linkage with
the environmental monitoring program.

Review the HVAC and room qualification, validation, calibration, maintenance

procedures and sample records where applicable.

Review deviations or investigations related to laboratory facility, cleaning and

sanitization or environmental issues.

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AUTOCLAVES

Review procedures and documentation for operation, cleaning, qualification, validation,

maintenance, controls and utilities (e.g. Clean steam) associated with the autoclaves.

Review logs and investigation of failed sterilization cycles and the disposition of the
contents of the failed cycles.

Inspect the autoclaves used for the sterilization of media.

(Autoclaves may lack the ability to displace steam with sterile filtered air. For sealed bottles of media,
this would not present a problem. However, for non-sealed bottles or flasks of media, non-sterile air has
led to the contamination of media. In addition, autoclaving less than the required time will also allow
media associated contaminants to grow and cause a false positive result. These problems may be more
prevalent in laboratories with a heavy workload.)

Check the temperature of the autoclave for a few historical cycles.

(Overheating can denature and even char necessary nutrients. This allows for a less than optimal
recovery of already stressed microorganisms. The obvious problem with potential false positives is the
inability to differentiate between inadvertent medium contamination and true contamination directly
associated with the sample tested.)

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ISOLATORS/GLOVE BOXES/LAMINAR FLOW HOODS

Review procedures and documentation supporting

- Operation
- Cleaning and sanitization
- Qualification/Validation
- Environmental monitoring
- Calibration and
- Preventive Maintenance including filter changes, glove integrity and integrity of
the closed areas.

Review investigations related to Isolator environment, change controls and for cause
repairs.

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INCUBATORS

Review the operation procedures, qualification, validation, calibration, cleaning,

sanitization, maintenance, change controls, alarms, and periodic monitoring schedules.

Review temperature and humidity logs for a selected time frame. Check for historical
log of excursions, and review investigations related to incubator performance.

Review how the temperature and humidity data is collected, e.g. location of probes,
data collection, archiving, and data reviews.

Check if any alarm system is installed and whether there is proper management
oversight of alarm conditions along with adequate corrective actions. Review
procedures and records regarding such activities.

If equipment is linked to a Building Management System (BMS), review the BMS

records for incubator records.

Review the overall condition of the incubator, its location, design, access, the way
materials are staged or stored within, identification of materials, general cleanliness,
state of repair, and any cleaning and sanitization schedules and corresponding
records.

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REFRIGERATORS

Review procedures and documentation supporting operation, cleaning and sanitization,

qualification, validation, environmental monitoring, calibration, and maintenance.

Review how the temperature data is collected, e.g. location of probes, data collection,
archiving, and data reviews.

Review investigations related to Refrigerator usage, change controls and for cause
repairs.

Review the overall condition of the refrigerator, its location, design, access, the way
materials are staged or stored within, identification of materials, general cleanliness,
state of repair, and any cleaning and sanitization schedules for the refrigerators.

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TEST INSTRUMENTS

Check calibration records, identification, and standards used for test instruments.

AUTOMATED IDENTIFICATION SYSTEMS

Review the operation procedures, qualification, validation, calibration, cleaning,

sanitization, maintenance, change control and periodic challenge testing of such
system.

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LABORATORY UTILITIES INCLUDING WATER SYSTEMS

Review operation procedures, diagrams, qualification and validation documents,

calibration records, cleaning and sanitization, maintenance, change controls, and
periodic monitoring of these systems.

Review investigations and excursions, related to such utilities and systems.

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MICROBIOLOGICAL TESTING OF NON-STERILE PRODUCTS

SPECIFICATIONS

Review microbial specifications for non-sterile products consistent with the USP
requirements or industry standards for the type of dosage forms. (Quantitative – total
counts, and qualitative - objectionable organisms, and endotoxins).

(The USP Microbiological Attributes Chapter <1111> states "The significance of microorganisms in non-
sterile pharmaceutical products should be evaluated in terms of the use of the product, the nature of the
product, and the potential hazard to the user." The USP recommends that certain categories be routinely
tested for total counts and specified indicator microbial contaminants. For example natural plant, animal
and some mineral products for Salmonella, oral liquids for E. Coli, for topicals - P. aeruginosa and S.
aureus, and articles intended for rectal, urethral, or vaginal administration for yeasts and molds. A
number of specific monographs also include definitive microbial limits.)

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RETESTS

Review instances where retests were performed.

(The results of initial test should have been reviewed and investigated. Microbiological contamination is
not evenly dispersed throughout a lot or sample of product and finding a contaminant in one sample and
not in another does not discount the findings of the initial sample results. Retest results should be
reviewed and evaluated, and particular emphasis should be placed on the logic and rationale for
conducting the retest.)

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IDENTIFICATION

Microbiological testing may include an identification of colonies found during the Total
Aerobic Plate Count test.

(The identification should not merely be limited to the USP indicator organisms. The importance of
identifying all isolates from either or both Total Plate Count testing and enrichment testing will depend
upon the product and its intended use. For example, if an oral solid dosage form such as a tablet is
tested, it may be acceptable to identify isolates when testing shows high levels. However, for other
products such as topicals, inhalants or nasal solutions where there is a major concern for microbiological
contamination, isolates from plate counts, as well as enrichment testing, should be identified.)

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MEDIA

Ensure that media used is appropriate for the test required. Review growth promotion
characteristics for all of the media used.

(Most microbiology laboratories employ enrichment media containing inactivators, such as Tween or
lecithin, to inactivate preservatives usually present in these types of product. This provides a better
medium for damaged or slow growing cells. Other growth parameters include a lower temperature and
longer incubation time (at least 5 days) that provide a better survival condition for damaged or slow-
growing cells. The selection neutralizing agents is largely dependent upon the preservative and
formulation of the product under evaluation. If there is growth in the enrichment broth, transfer to more
selective agar media or suitable enrichment agar may be necessary for subsequent identification.)

If possible, review or witness analyses being conducted and inspect the plates and
tubes of media being incubated (caution should be exercised not to inadvertently
contaminate plates or tubes of media on test).

(Be particularly alert for retests that have not been documented and "special projects" in which
investigations of contamination problems have been identified. This can be evaluated by reviewing the
ongoing analyses (product or environmental) for positive test results. Request to review the previous
day's plates and media, if available and compare your observations to the recorded entries in the logs.)

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STOCK CULTURES & GROWTH PROMOTION STUDIES

Review how stock cultures of reference organism are procured and stored. Review the
procedures for stock culture usage.

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REAGENTS

Review the laboratory reagents storage, handling and identification. Identify how
obsolete reagents and dilutions are disposed.

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SAMPLING

Review sampling procedures including, collection techniques, sampling containers,

storage areas, storage conditions, time limits, transportation methods, receiving
process (in the laboratory), staging and processing.

Assure that inhibitory substances in samples are neutralized prior to testing and
sample integrity is maintained.

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METHODOLOGY AND VALIDATION OF TEST PROCEDURES

Determine the source of test procedures. (e.g USP, BAM and other microbiological
references).

Evaluate the methodology for microbiological testing. (Test methods must be able to
identify the presence of organisms such as Pseudomonas cepacia or other
Pseudomonas species that may be objectionable or present a hazard to the user.
Where applicable, compare the method being used against the one submitted in the
application.)

Verify that the laboratory has the equipment necessary to perform the tests and that
the equipment was available and in good operating condition on the dates of critical
testing. Per the USP, an alternate method may be substituted for compendial tests,
provided it has been properly validated as giving equivalent or better results.

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MEDIA CHALLENGE

Review whether the laboratory conducts periodic challenge of prepared media with low
levels of organisms including USP indicator organisms as well as normal flora.

(The capability of the media to promote the growth of organisms may be affected by the media
preparation process, sterilization (overheating) and storage. These represent important considerations in
any inspection and in the good management of a microbiology laboratory.)

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DATA STORAGE

Review procedure and policies or data recording, documentation practices, storage

and archiving.

Evaluate the test results that have been entered in either logbooks or on loose
analytical sheets. Pay particular attention to ensure data integrity. When summaries of
this data are not available, review enough data to construct their own summary of the
laboratory test results and quality control program.

Where an automated microbial system for the identification of microorganisms is used,

review logs of such testing.

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CONTRACT TESTING LABORATORY

Evaluate the firm’s program to audit the quality of the laboratory work performed by
outside contractors.

Determine the specific instructions given to the contractor. Review if there are any
Quality Agreements between the contract laboratory and the firm and if not how the
Quality System and compliance requirements of the firm monitored and enforced.

Analytical results, particularly for those articles in which additional or retesting is

conducted, should be reviewed. Test reports should be provided to the manufacturer
for tests conducted.

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STERILITY TESTING

Evaluate the facility for sterility testing.

(Per USP -"The facility for sterility testing should be such as to offer no greater a microbial challenge to
the articles being tested than that of an aseptic processing production facility". Proper design would,
therefore, include a gowning area and pass-through airlock. Environmental monitoring and gowning
should be equivalent to that used for manufacturing product.)

Perform actual observation of the sterility test, if possible.

(Make the observations in a manner that will create the least amount of disruption in
the normal operating environment.)

Examine the use of negative controls during sterility testing.

Request lists of test failures to facilitate review of production and control records and
investigation reports.

Review records of initial positive sterility test results.

(Good practice for such testing includes the use of known terminally sterilized or irradiated samples as a
system control. Alternatively, vials or ampules filled during media fills have also been used. Be
especially concerned about the case where a manufacturer of aseptically filled products has never found
an initial positive sterility test. While such situations may occur, they are rare. The absence of initial
positives may indicate that the test has not been validated to demonstrate that there is no carryover of
inhibition from the product or preservative.)

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STERILITY TESTING

Inspect robotic systems or isolation technology, such as La Calhene units used for
sterility testing. These units allow product withdrawal in the absence of people.

(If an initial test failure is noted in a sample tested in such a system, it could be very difficult to justify
release based on a retest, particularly if test controls are negative.)

Evaluate the time period used for sterility test sample incubation.

(Most laboratories use a period of 14 days incubation.)

Bacteriostasis and fungiostasis testing must be performed. These test results must be
negative otherwise any sterility test results obtained on the product may not be valid.

Media fill, environmental, sterility test results and other data should be reviewed to
assure the absence of slow growing organisms.

Where applicable, compare the methods being used for incubation to determine if they
conform to those listed in approved or pending applications.

*This check list includes certain material published in the US FDA Guidance for
Inspection of Microbiology Laboratory for Non-Sterile Products.

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