Microbiological Best Lab

Practice &
Environmental Monitoring
Romauli Tryana Sinaga
24 October 2020
Pharmaceutical Industry
• Good Manufacturing • GMP is integrated into all sequences of
production from the raw material, the
Practice (GMP) is a premises design, to the equipment
process that ensure that used.
products that are • Also the personnel training and
produced by hygiene are promoted since it is the
principal contributor to GMP.
pharmaceutical
companies are of quality
Microbiology, what I need to know?
• Microbiology is the crucial section of Sterile Pharmaceutical
products also it is very Important in Non- Sterile Products.
• Microbiology sections can be divide in following sections:
1. Sterility Testing
2. Bacterial Endotoxin Testing
3. Microbial Examination of Non-Sterile Products
4. Antibiotic Potency Testing
5. Environmental Monitoring
6. Methods of Sterilization
7. Antimicrobial Effectiveness Testing
USP 〈1117〉 MICROBIOLOGICAL BEST
LABORATORY PRACTICES
1. Media preparation and Quality control
2. Maintenance of Microbiological Cultures
3. Maintenance of Laboratory Equipment
4. Laboratory Layout and Operations
5. Training of Personnel
6. Documentation
7. Maintenance of Laboratory Records
8. Interpretation of Assay Result
Media preparation and Quality control
• Microorganism needs nutrients and certain environmental conditions
for their growth and reproduction.
• In a laboratory, this growth requirement are met artificially by means
of culture media.
• The quality of work in a microbiological laboratory depends on
the quality of the culture media.
• Media preparation, proper storage, and quality control testing can
assure a consistent supply of high quality media.
Media preparation and Quality control
• Accurate weighing of dehydrated
components
• The use of high quality (USP Purified)
water,
• Completely dissolving the dehydrated
media or individual ingredients,
• The need to control the heating of the
media to avoid damaging heat-labile
components of the media.
• Some recommendations on the
labeling and packaging of media are
also provided.
• Pre-incubation
• Growth Promotion Test (GPT)
Maintenance of Microbiological Cultures

• Safeguarding the stock cultures is

the most important component of a
successful microbiology laboratory.
• The use of an incorrect strain in a
compendial test could bring the
results of weeks or months of work
into question.
• A careful lab will safeguard the purity
and identity of their stock cultures by
limiting the potential for “drift” due to
excessive transfers.
Maintenance of Microbiological Cultures
Maintenance of
Laboratory Equipment

• Microbiology laboratory =
mini scale of production
• Most lab equipment in the
microbiology laboratory is subject
to the standard validation
practices of IQ, OQ, and PQ.
• Periodic calibration/maintenance
may be required for the
particular equipment based on its
nature,
• And performance verification
checks should also be
performed regularly.
Laboratory Layout and
Operations

• Understand or plan for the

separation of samples from a
microbiological perspective.
• The success of a laboratory can be
enhanced by the thoughtful separation
of samples likely to have contamination
from those that are expected to be
sterile.
Training of Personnel
• What should be common sense in recommending that
microbiologists and managers in the pharmaceutical support lab
should have academic training in microbiology or allied health
sciences.
• It recommends that the SOP system should be comprehensive and
serve as basis of the training program.
• Performance assessments to be done periodically and
should demonstrate competency in core activities of the lab.
Documentation and Maintenance
of Laboratory Records

• Blue ink
• Formal approved form
• Data Integrity (ALCOA)
• Archiving
Interpretation of Assay Result
• OOS (Out of Specification) Investigations
• We are dealing with such low numbers on plates (frequently less than
20 CFU/plate) and the real opportunities for human error in tests
that may run over a month to completion, the microbiologist must
always be aware of the role that random chance has in the data and
be on guard against over-interpreting the results of a study.
• A guide to developing methods of investigating test failures.
• The difference between a test that has failed, a test that should be
invalidated and a test that should be repeated for confirmation.
Sample for microbiology testing

Raw material solid/powder

Raw material liquid
Water (softened water, purified wa
Finished good (solid, liquid, liquid in
A pharmaceutical
microbial laboratory
mostly associated
tests.
• Sterility Test
• Quantification of the
microorganism (Total Place
Count)
• Detection and
identification of pathogenic
bacteria
• Endotoxin Test
Sterility Test
• Sterile finished good
• Filtration method, broth media, qualitative → Pass or Fail
• 14 days incubation
• Sterility test Isolator
Endotoxin Test
• Sterile finished good, raw material, WFI (Water for Injection)
• Few hours testing
Environmental Monitoring (EM)
• The manufacture of pharmaceutical products must be performed
under controlled conditions.
• Monitoring is, therefore, an important part of proving that the
manufacturing process is under control, and in aseptic production it
plays a particularly important role.

• What we measure, why and how?

• EM plan, risk based approach
Guidelines and regulations for microbiological
monitoring in sterile & non-sterile production
• PIC/S PE 009-13 Guide to Good Manufacturing Practice for Medicinal Products Annexes These
guidelines also lay down the permitted limits, both for particles and for microbiological contamination
• EU GMP guideline, Annex 1: Manufacture of Sterile Medicinal Products
• ISO 13408-1 Aseptic processing of health care products ISO 14644-1 Cleanrooms and associated
controlled environments: Classification of air cleanliness by particle concentration
• ISO 14698-1 Cleanrooms and associated controlled environments: Biocontamination control
• FDA, Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing - Current Good
Manufacturing Practice • USP Chapter <1115> Bioburden Control of Nonsterile Drug Substances and
Products
• USP Chapter <1116> Microbiological Control and Monitoring of Aseptic Processing Environments
• PDA Technical Report No. 67 - Exclusion of Objectionable Microorganisms from Nonsterile
Pharmaceuticals, Medical Devices, and Cosmetics. 2014: Parenteral Drug Association, Inc.
• PDA Technical Report No. 13 (Revised) - Fundamentals of an Environmental Monitoring Program.
Parenteral Drug Association, Inc., 2014
• ANVISA Quality Guide for Air Treatment and Environmental Monitoring Systems in the Pharmaceutical
Industry (2013)
Environmental Monitoring (EM)
EM based risk approach
Personnel
flow
•Sampling points in sufficient detail Contamination
Material flow
vectors
•Sampling frequencies that provide
meaningful results
•Sampling time (e.g. during filling or Historical data
Difficult to
disinfect
at the end) Sampling
•Duration of sampling Point
•Sampling size (e.g. area of the Selection
Cleaning &
Product risk
imprint, air volume) disinfection
•Sampling equipment
•Sampling techniques Size of the Product
•Warning and action levels area
Activity
exposure
•Corrective and preventive actions if performed
warning and action levels are
exceeded
Method of Sampling
• Particle count
• Airborne
• Air sampler
• Settle plate
• Surface
• Contact plate
• Swab
• Finger print
Particle Count, Air sampler, settle plate
 Surface
monitoring
Environmental Monitoring (EM)
• Large amounts of data are generated by the broad-based monitoring
of air, surfaces and, if necessary, water, which in critical areas may
even be carried out as permanent monitoring or with a high
sampling frequency.
• Trend data analysis for continual improvement, requalification etc.
Thank you

Any questions?