This Sop Describes the Procedure for Effectiveness of Antibacterial preservative.
This Sop is Applicable to the Procedure for Effectiveness of Antibacterial preservative.
Head Quality Control.
- Test organisms:
- The following test organisms are used in the test.
- Candida albicans,
- Aspergillus niger
- Escherichia coli
- Pseudomonas aeruginosa
- Staphylococus aureus
- *Instead of Candida albicans, Zygosccharomyces rouxii.
- In order to prevent any
phenotypic changes in the strains used, the organisms used in the test
should not be more than 5 passages made from the original culture. One
passage is defined as inoculation and growth of the organisms
from existing culture to a fresh medium.
NOTE- All the media used in the tests should be tested for growth promotion.
- Preparation of inoculums:
- Grow each of the bacterial species separately in Casein soyabean digest agar and incubate them at 30° to 35° for] 8 to 24 hours. Grow Candida albicans on Sabouraud dextrose agar and incubate at 20° to 25° for atleast 48 hours. Grow Aspergillus niger on Sabouraud dextrose agar at 20° to 25° for 5-7 days. After incubation, harvest the growth and resuspend each of the organisms separately in sterile to obtain a microbial count of lx108 cfu per ml. To suspend spores of Aspergillus niger 0.05per cent polysorbate 80 may be added to the saline. Use suspension of these organisms within 2-4 hours. The suspension may be stored at 4° to 8° for a validated period of time. Remove immediately a suitable sample from each suspension and determine the number of cfu per m!. in each suspension by pour plate method or filtration method. This value serves to determine the inoculum concentration and the baseline to use in the test.
- If sufficient volume (atleast 20 ml) of product is available in each container and the product container can be inoculated aseptically then the test can be conducted in five original containers of the product. If filled volume is less, or the container cannot be inoculated aseptically then transfer (atleast 20 ml) the product in each of five suitable sterile containers. Inoculate each container with one of the prepared and standardized inoculum in such a way that after inoculation the final concentration of the organisms remains between Ix 105 and Ix 106 cfu per ml and the volume of the inoculum does not exceed Iper cent of the volume of the product. The initial concentration of the viable organisms in each test preparation is estimated based on the concentration of the microorganisms in each of the standardized inoculum as determined by the pour plate method or membrane filtration method. Incubate the inoculated containers at room temperature.
- Determine the viable count by plate-count method at 7, 14, and 28 days subsequent to the inoculation. Record any changes observed in the appearance at these intervals. From the calculated concentration of cfu per rn1present at the start of the test, calculate the percentage of reduction in cfu per ml for each organism at the stated test intervals and express the changes in terms of percentage of initial concentration.
- The preservatives are considered to be effective if:
- For oral preparations: (a) The concentration of the viable bacteria are not more than 10 per cent of the initial concentration at 14 days and there is a further decrease in count at 28 day. (b) There is no increase in yeast and mold count at 14 and 28 days from the initial count.
Trainer : Manager – Quality Control
Trainees : Staff of the microbiology departments
Master Copy : QA Department
Controlled Copy : Microbiology Department
Display Copy : Microbiology Department (If Required)
|Sr. No.||Title||Annexure No.|
|01.||Record of Effectiveness of Antibacterial preservative||NA|
|SOP||Standard Operating Procedure|
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- REVISION HISTORY OF CHANGE:
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|02||NA||SOP Periodical Review||NA|