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Learn how to validate the method of sterility testing by membrane filtration method.
1.0 INTRODUCTION
When any test for sterility is initially carried out for any product, it is necessary to validate the test method used, by recovery of a few number of
microorganisms in the presence of the product.
2.0 OBJECTIVE
The Objective of this validation is to establish documented evidence that the test for sterility by membrane filtration method will produce the consistent
results, when analyzed as per the standard Operating Procedure.
3.0 SCOPE
This protocol is applicable for Sterility test of the products.
4.0
RESPONSIBILITIES
S. No. Responsibilities Name of the
Department
1 Preparation of Validation Protocol Quality Control
2 Execution of Protocol Quality Control
3 Approval of Protocol and Report Quality Assurance
4 Final System Approval Head QA / QC
5 Review and maintenance of Report Quality Control
5.0 PREREQUISITES
In order to efficiently conduct validation of the Sterility Test by Filtration method, ensure that the following requirements are fulfilled.
1. Validated Aseptic facility to carry out the Sterility test Validation
2. All Equipments to be used for Sterility test validation are qualified and operational SOPs established and followed.
3. All the equipments and culture media required for the validation of sterility test should be sterile.
4. Trained personnel for conducting Sterility test and its Validation.
5. Membrane filter: Sterile individually packed cellulose nitrate or cellulose acetate, 47 mm diameter, average pore size 0.45m.
6. Sterilized filtration assembly, forceps and scissor
7. Sterile 70% IPA solution
6.0 EQUIPMENT / SYSTEM DESCRIPTION
1 Location Sterility Room of Quality control department
2 Equipment The equipment covers the Stainless steel filtration assembly with
manifolds, Vacuum pump, Autoclave, Hot air Oven, Laminar air
flow , and Incubator
7.0 IDENTIFICATION OF CRITICAL CONTROL / MONITORING PARAMETER
Before proceeding for sterility test Validation, following parameters to be checked
7.1 Each lot of dehydrated media used for preparation of sterility test medium must be tested for its growth promoting qualities as per SOP.
7.2 During validation carry out environmental monitoring by settle plate Plate Exposure and Personnel monitoring by finger dab and swab method as per
SOP.
7.3 If any cfu observed during monitoring on LAF and Finger dab, all cfu must be identified up to species level.
8.0 SAMPLING DETAILS
1 Carry out the sampling of three consecutive batches from various sites throughout the sterilizer load.
2 Immediately carry out the leak testing and visually examine the bottles for any leakage or any extraneous particles.
Table I
Sr. No Positive control Fluid Thioglycollate Soyabean Casein Digest
Medium Medium
1 Positive Control S. aureus NCIM 2079 C. albicans NCIM 3471
I
2 Positive Control Ps. aeruginosa NCIM A. niger NCIM 1196
II 2200
3 Positive Control B. subtilis NCIM 2063 Environmental Flora EF II Acceptance Criteria
III 1. If the conspicuous growth is observed within 3 days for
4 Positive Control Environmental Flora EF I bacteria and 5 days for fungi, and the growth of each
IV challenge microorganisms in the Positive Product control
containers are visually comparable to the growth in the
positive control and there is no growth in negative control & negative product control, the product possess no antimicrobial activity under the condition of
the test or such a activity has been satisfactory eliminated. The test for sterility may be carried out routinely without further modifications.
2. If the conspicuous growth is not observed within 3 days for bacteria and 5 days for fungi, or growths of each test organism in the Positive Product
Control containers are visually not comparable with positive control containers respectively, the product possesses antimicrobial activity that has not been
Control containers are visually not comparable with positive control containers respectively, the product possesses antimicrobial activity that has not been
satisfactory eliminated under the conditions of the test. Modify the conditions in order to eliminate the antimicrobial activity and repeat the validation test.
i.e. by using additional washes
Negative Product Control Test:
The result of negative product control test facilitates the interpretation of sterility test results, particularly when used to declare a test invalid because of
contamination in negative product control. The essential element of the negative control is to simulate the testing method.
B. TEST FOR ANTIMICROBIAL ACTIVITY
Objectives:
The test is performed to ensure that, the absence of Antimicrobial Activity under the experimental conditions.
Procedure:
Step Product Control Positive Control Negative Control
No
1 Rinse the membrane with approx NA Negative Control I
15 ml of sterile peptone water Rinse the membrane with
approx 15 ml of sterile
peptone water
2 Select 20 bottles randomly and NA Rinse the membrane with
pull the half content full content 100 ml sterile peptone
of container in case of SVP into a water
filter holder & start the filtration
Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widelyread pharmaceutical
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