Points to Consider when Validating

your Sterility Testing Canisters

Sterility testing is an integral part

of all pharmaceutical microbiology
laboratories and is designed to
detect the presence of viable
microbial contaminants in sterile
pharmaceuticals. Being a method
based on the evaluation of
microbial growth, it is crucial to
distinguish between true product
sterility and a false negative
(Aseptic Guideline 2004).
Certain ingredients used in the
formulation of drugs can possess
innate anti-microbial properties
and prevent a sterility test from
reliably reporting on the presence
of viable microorganisms..

Validation is therefore performed

for all new product formulations,
whenever there is any change
made in product formulation or if
there are changes in experimental
conditions. This includes selecting
or making a change between
a primary | secondary supplier of
your sterility testing equipment.
Even in the absence of a change,
it is recommended to routinely
revalidate all processes on a
regular basis.

Sterility Testing
This involves the continual collection, evaluation and 4. Reason for Validation
documentation of data. As a general rule, it is advisable to All critical materials and assays used in the manufacture
seek guidance and feedback from local or international and release of parenteral products must be validated for
regulatory bodies or advisors on the proposed methodology suitability. Validation of Sartorius Sterisart® canisters should
early in the process and prior to undertaking a validation be performed as indicated by the appropriate
exercise to ensure these will comply with their requirements. pharmacopoeia, regulatory guidelines, or both.

There are several detailed guidelines for sterility testing, Manufacturers may want to consider validating materials and
besides the pharmacopeial chapters. We have compiled consumables from alternate suppliers for mission critical
the following points to be considered during the testing consumables to provide protection against process
validation | revalidation of your sterility testing canisters. interruptions due to supply outages. These outages could be
due to logistics issues, where the consumable | test item is
not available locally, or if there is a failure in the manufacturing
process. Examples of causes for these outages include fire,
earthquake, or other natural or man-made disaster at the
factory manufacturing the test consumable, lack of raw
material(s) required to manufacture the test consumable,
logistics issues caused by a break-down in the logistics chain
due to strike action, a pandemic, or a similar interruption
to freight.

5. Revalidation Criteria
Test methods should be revalidated if there is a change in
manufacturing procedures, testing procedures or any of the
test items, including the consumables or growth media used.
The PIC/S 11.6.2.4 and the TGA guidelines on sterility testing
(407) also recommends revalidating test methods every 12
months, although this is not a pharmacopoeial requirement.
Validation of Sterisart® Sterility Revalidation may be required if a manufacturer of a
Testing Canisters consumable or media changes the construction, materials
used, functionality of the consumables, or the formulation or
Sterility testing canisters must be compliant with the ingredients in a media type.
pharmacopoeia guidelines used in the facility, and a
manufacturer’s validation guide should be available. 6. Responsibilities
Ultimately, methods validation studies should demonstrate The head of Quality Assurance is responsible for approving
that the method does not provide an opportunity for false the validation protocol, and for accepting changes to the
negatives (Aseptic Guideline 2004). The following points testing procedure once equivalence of the second supply
should be considered when selecting or making a change to has been demonstrated. The head of the Microbiology
suppliers of sterility testing canisters or any other critical Laboratory is responsible for overseeing the implementation
component of this validation protocol, and the sterility test team is
of manufacture or testing of a product. responsible for performing the associated tasks. Sartorius will
provide its in-house validation document. This document
1. Approval demonstrates that the Sterisart® canisters meet or exceed
This Sterility Testing Canister Validation Protocol should the requirements for use in a compendial sterility test in terms
be reviewed by the head of Microbiology or an authorised of the materials and methods used in their manufacture,
QC microbiologist and approved by the head of Quality assembly, packaging, and sterilization.
Assurance or their designated authority.
7. Reference Documents
2. Objective In-house standard operating procedures that comply with
The object of this protocol is to validate Sartorius Sterisart® relevant pharmacopoeia, cGMP and PIC/S guidelines, and
Canisters as approved canisters for use during quality control any other appropriate guidelines or regulations should be
and lot release testing in this facility. referenced to perform this validation protocol. Validation
experiments will be based on the current, validated work
3. Scope methods and SOPs in the first instance, and varied as
This protocol is relevant for the sterility testing team in the required to achieve product validation. These internal
microbiology laboratory of the quality assurance department reference documents may need updating to include any
of the facility.
necessary variations from the current methodology required sets per batch of product tested. Negative controls (one
to successfully utilize the Sterisart® canisters. Established canister set) and negative product controls (one canister set
change-management procedures for updating documents per product) should also be conducted. While a definitive
should be followed if changes are required. number of batches of product are no longer specified for
assay validation, regulators expect manufacturers to use a
8. Procedure science-based approach to determine how many batches will
The following is a general overview of the procedure involved be used during validation or revalidation, and to have sound
in the process of validating a sterility test, and is by no means rationale for this decision. Consequently, a typical validation
exhaustive. Please refer to the appropriate pharmacopoeia experiment will consist of eight or more canister sets as
document or local authority guidelines for a more complete defined in Table 1.
description.

For new products, validation of the inactivation of product

ingredients having anti-microbial activity, or rinsing them
from the membrane will be required. For most re-validations
of existing assays due to any of the changes listed above,
existing inactivation, filtration, and rinse parameters should
be suitable, but must be re-validated using bacteriostasis
and fungistasis tests.

This should be performed generally by spiking the final rinse

solution with <100 CFU of test organisms. In the case of
products having no demonstrable anti-microbial activity,
and that require no rinsing of the membrane (for example
isotonic saline solutions), the product itself may need to be
spiked. Growth promotion tests (positive controls) should
be performed alongside sterility testing to validate the
growth of test species in this assay. Bacteria should grow Negative product controls should have all filtration and rinse
within 3 days and fungi within 5 days for valid results, and steps performed apart from the actual product filtration,
there should be little visible difference in microbial growth which is substituted by product or simulated product of
between the positive controls and the test canisters. known or undoubted sterility. Negative product controls
Occasionally, there are regional differences in should be exposed to a terminal sterilisation process, such as
recommendations and practices. For instance, the TGA exposure to steam sterilisation, gamma-irradiation etc., and
guideline specifies that growth promotion tests should be be packaged in a similar manner to the test sample.
performed after 14 days on un-spiked sterility test samples to Alternatively, distilled water in the same or similar container
show the media are still capable of supporting growth. could be used. Growth results of all canisters should be
Bacteria should grow within 3 days and fungi within 5 days for recorded as pass | fail. Photographic records are desirable. Where
valid results. necessary, inactivation and rinse steps may need to be
re-optimised to achieve the desired growth results in the
Growth promotion tests (positive controls) will require three bacteriostasis | fungistasis tests. These should be recorded
canister sets for confirming the growth rates and patterns as deviations from the existing test method.
of the six standard species in the assay being validated, and In urgent situations, concurrent validation of alternative
bacteriostasis and fungistasis tests will require three canister sterility test canisters can be performed as part of routine testing.

Canister set Contents TSB inoculum FTM inoculum

1 Growth promotion assay (positive control) Bacillus subtilis Clostridium sporogenes

2 Growth promotion assay (positive control) Candida albicans Staphylococcus aureus

3 Growth promotion assay (positive control) Aspergillus niger Pseudomonas aeruginosa

4 Bacteriostasis | fungistasis test (per batch) Aspergillus niger Clostridium sporogenes

5 Bacteriostasis | fungistasis test (per batch) Candida albicans Staphylococcus aureus

6 Bacteriostasis | fungistasis test (per batch) Aspergillus niger Pseudomonas aeruginosa

7 Negative product control None None

8 Negative control None None

Table 1. Species inoculation scheme for sterility canister testing

Preferably, this should run alongside existing sterility tests, 10. Conclusions
using consumables from the current supplier(s) to The conclusion should include the overall results of the
demonstrate equivalence to existing methods and validation process indicating if validation passes or fails for
consumables. each product and should note any changes required to the
test method for the tests to successfully pass the assessment
criteria.

11. Report
A report including the raw data for each product evaluated
should be attached as annexure. Data should be compiled
by the operator performing the validation and should be
checked by the head of Microbiology or their designated
authority.

12. Report Approval

Reports should be reviewed by concerned departments
and approved by the head of Quality Assurance or their
designated authority. Report approval shows that the
validation was completed successfully and according to
the validation protocol.

References
U.S. Pharmacopeia. USP <71> Sterility Test
European Pharmacopoeia. Ph. Eur. 2.6.1 Sterility
9. Deviations Japanese Pharmacopoeia. JP 4.06 Sterility test
Any and all deviations from the written procedure occurring World Health Organization (WHO); 3.2 Test for sterility
during the validation activity should be recorded.
Any deviation occurring in inactivation or rinse steps due to TGA guidelines for sterility testing of therapeutic goods, 2006
changes required to achieve acceptable growth rates in the 21 CFR 610.12 – General Provisions
bacteriostasis | fungistasis tests in a revalidation should be PIC/S PI 012-2 Recommendation on Sterility Testing
recorded. FDA Aseptic Guideline (Sterile Products Produced by
Aseptic Processing, 2004)

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Status: 07 | 2020