Sr. No. |
Description |
1.0 |
Protocol
Approval |
2.0 |
Objective |
3.0 |
Scope |
4.0 |
Validation
Description |
5.0 |
Responsibilities
and Identification of Execution Team |
6.0 |
Test
Procedures |
7.0 |
Recording
of Observations |
8.0 |
Discrepancy
and Corrective Action Report |
9.0 |
The compilation,
Review, and Summary Report |
10.0 |
Appendix |
11.0 |
Revalidation
Criteria |
Department |
|
Quality
Control Microbiology |
Execute,
Participate and provide the necessary support for the validation activity. Preparation
and review of the validation Report of the documents and its compliance to meet
the acceptance criteria of the protocol. |
Quality
Assurance |
Monitoring
the validation activities, compilation, review and authorization of the method
validation report and its compliance to meet the acceptance criteria of the protocol. |
- Sterile product sample.
- Sterile distilled water.
- Sterile Molten Soyabean Casein Digest Agar.
- Poured SCDA plates.
- Sterile forceps.
- Sterile membrane filtration units.
- Sterile membranes.
- Vortex Mixer.
- 0.1 % peptone water.
- Sterile Fluid Thioglycollate Medium.
- Sterile Soyabean Casein Digest Medium.
- Cultures required suspensions of Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium sporogenes, Bacillus subtilis, Candida albicans, and Aspergillus Niger.
- Preparation of 10 –100 cells/ml: Prepare the 10 – 100 cells/ml solution as per the SOP.
- To Determine the Sterility of the Product.
- After sampling, the sterile product sample will bring to the microbiology lab.
- The microbiologist will place the sterile product sample within the pass box after decontaminating the outer surface of the pouch with disinfectant solution.
- Enter the sterility testing room as per the SOP for sterile area entry and exit procedure for the sterility testing lab.
- After entering the sterility testing room, clean the LAF surface with 70 % filtered IPA
- Place the Sterile product Samples within the LAF.
- Perform the sterility test of the sample by following the procedure described in SOP for Sterility Testing.
- Operate the sterility test apparatus as per SOP.
- After performance Incubate Soyabean Casein Digest Medium at 20 – 25°C for 14 days and Fluid Thioglycollate Medium at 30 – 35°C for 14 days.
- Observe the medium tubes/canisters daily for any growth.
- Record the observations daily in the format given as annexure-1.
- To Determine the Bacteriostasis and Fungistasis nature of the Product.
- Perform the whole exercise under the LAF in the Sterility testing area.
- Take the sterile sample as per the SOP for the Sampling of Sterile Raw Material and dissolve the sample in 200 ml of 0.1 % peptone water.
- Perform the sterility test for the above sample by following the procedure described in SOP for Sterility Testing.
- Rinse the membrane with 2 X 100 ml portions of sterile 0.1 % peptone water.
- Inoculate the final rinse with 10 - 100 CFU/ mL of the test microorganisms.
- After filtration aseptically transfer the membrane onto the SCDM / FTGM depending on the organism being tested.
- Perform the whole exercise with all the organisms listed below.
- Note down the results in Annexure-1.
- Positive Control:
- Filter 2 X 100 ml portions of 0.1 % peptone water through a 0.45μ filter membrane.
- Finally, rinse the membrane with the third 100 ml 0.1 % sterile peptone water inoculated with 10 – 100 cells/ml of any culture listed above.
- Repeat the above exercise with all the other organisms.
- Inoculate the membrane into a medium.
Sr. No. |
Medium |
Organisms |
Strain Number |
01 |
Fluid
Thioglycollate |
Staphylococcus
aureus |
|
Pseudomonas
aeruginosa |
|
||
Clostridium
sporogenes |
|
||
02 |
Soyabean
Casein Digest |
Bacillus
subtilis |
|
Candida
albicans |
|
||
Aspergillus
niger |
|
- Negative Control
- Filter 3 X 100 ml portions of 0.1 % peptone water through 0.45μ filter membrane.
- Cut the membranes into two halves and inoculate each half separately into a specified medium.
- Incubation of Tubes
- Incubate test for sterility sample tubes of Fluid Thioglycollate Medium at 32.5 ± 2.5°C and for Soyabean Caesin digest Medium incubate at 22.5 ± 2.5°C for 14 days.
- The negative control should be incubated for 14 days, observed daily and recorded.
- Incubate the Positive control tubes for 3 days in case of bacteria and 5 days in case of fungi.
- Acceptance Criteria
- The growth of each test organism in the test sample (with the product) is visually comparable to the growth of the positive control.
- The negative control should not show any growth during the 14 days of incubation.
Abbreviation |
Definitions |
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- Preparation Of Culture Suspensions.
- Sterility Testing.
- Growth Promotion test.
- Sterility Testing Area, Entry And Exit Procedure SOP.
- Operation Of Equinox; Make: Millipore. SOP.
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