EXERCISE 7: STERILITY TESTING

Aseptic processing
Factors the influence the outcome - Designed to prevent the introduction of viable
Environment microorganisms into separately sterilized
- Facility design materials during their assembly into a sealed
- Personnel traffic flow sterile package (primary
- HVAC
- Personnel practices and training Terminally-sterilized products
- Cleaning and maintenance - Lowest-risk category of sterile pharmaceutical
- Qualification products
- Personnel hygiene o Products that can withstand heat and will
- Decontamination not disintegrate in presence of heat
- Utility systems - Sterilization process imparts quantifiable safety
- Effects from adjacent areas level achieved by delivering measurable physical
conditions that corresponds to microbial lethality
Equipment - Probability of the terminal sterilization process
- Equipment design generating a nonsterile unit of ≤ 10-6
- Personnel practices and training
- Cleaning and maintenance Aseptic processing
- Procedures - Each component is individually sterilized, or
- Qualification several components are combined with the
- Sterilization resulting mixture sterilized
- Area equipment - Preparation of solution (Filtered through
sterilizing filter then filled into sterile containers)
Product - Each component is individually sterilized, and
- Storage conditions each process is under aseptic technique
- Personnel practices and training
- Product and materials Terminally-sterilized products
- Depyrogenation - For products that can withstand the steam
- Sterilization sterilization cycle
- Product and material flow - Assure sterility of finished goods
- Validation - Product is sterilized in its final container
- Ionizing radiation
Sterile products - Steam sterilization (autoclave)
- Process is designed to prevent the introduction of - Dry heat sterilization (Oven)
viable microorganisms into/onto separately
sterilized materials during their assembly into a Sterility Testing
sealed sterile package - Applied to substances, preparations, or articles
o Each package should be sterilized which are required to be sterile
▪ Primary packaging – most - Indicated that no contaminating microorganism
important, direct contact to product has been found in the sample examined under the
o Steps: Aseptic Techniques should be conditions of the test
observed - Carried out under aseptic conditions
- Injections, ophthalmic preparations, irrigation - Membrane filtration method
solutions (used to wash wounds), haemodialysis o Filters with nominal pore size of n.m.t.
solution (solution used by renally impaired 0.45 mcm
patient) o Cellulose nitrate filters
- Two Categories: ▪ Aqueous, oily, and weakly
- Terminal sterilization alcoholic solutions
o Can be sterilized in final container o Cellulose acetate filters
- Aseptic processing ▪ Strongly alcoholic solutions
o Cannot be terminally sterilized and must - Direct inoculation method
be aseptically prepared (in every step)
EXERCISE 7: STERILITY TESTING

- Before preparations are labelled as sterile

products, they should have formulated and sealed
under aseptic conditions
- They should also have underwent appropriate
physical or chemical sterilization method prior to
sterility testing

Sterility Testing
- Culture media designed for sterility testing
- Fluid Thioglycollate medium (FTM)
o Anaerobic bacteria
o Aerobic bacteria
o Thioglycolate and cysteine in the medium
reduces the oxygen to water, this
decreases the level of oxygen and
promotes the growth of anaerobes Growth Promotion Test
- Soybean-Casein Digest medium (SCD) - If microorganisms are able to grow
o Fungi o It means that the sample doesn’t inhibit
o Aerobic bacteria their growth and sterility test sterility
o Highly nutritious testing can proceed

Method Suitability Test (Method Validation)

- Conducted to demonstrate the applicability of the
method for detection of microbial contamination
in the test product
- Before beginning with the sterility test, a Method
Suitability Test should take place
- This is performed when first, a test for sterility has
to be carried out on a new product or whenever
there’s a change in the experimental conditions of
the test
- Suitability tests:
- Growth promotion test Suitability of Test Method
o To demonstrate that the medium - If clearly visible growth is not obtained in the
supports growth and has the ability to product to be tested
detect organisms in the presence of the o It means that the product possesses
test sample antimicrobial activity that has not been
- Suitability of test method eliminated under the conditions of the
o Based on sample characteristics and test
required microbial limits o Modification of the condition is
recommended in order to eliminate the
antimicrobial activity and repeat the
suitability of test method
- If no evidence of microbial growth is found
o Then the product complies with the test
for sterility