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 |
Compilation,
Review and Summary Report |
10.0 |
Appendix |
11.0 |
Revalidation
Criteria |
Department |
|
Quality
Control Microbiology |
Execute,
Participate and provide 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 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 the 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 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 incase of fungi.
- Acceptance Criteria
- The growth of each test organisms 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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