Standard operating to test the disinfectants used for cleaning in pharmaceutical company for its efficacy using the different types of microorganisms.
1.0 OBJECTIVE
To determine the efficacy of disinfectants.
2.0 SCOPE
This procedure is applicable for all the disinfectants used for disinfection at manufacturing plant.
3.0 RESPONSIBILITY
QC-Microbiologist
4.0 ACCOUNTABILITY
Head QC
5.0 ACCOUNTABILITY
Head QC
PROCEDURE
5.1 Principle: The disinfectant efficacy is based on the ability to reduce the microbial population by 90% within 15 minutes. Here the known concentration of microorganisms is mixed with the disinfectant of respective concentration. And the recovery of the microorganisms is checked at 5 minutes, 10 minutes, and 15 minutes.
5.2 Requirements
Disinfectant solution
Normal saline.
SCDA & SDA
Sterile purified water
Microbial culture suspension of,
Escherichia coli ATCC 8739
Pseudomonas aeruginosa ATCC 9027
Staphylococcus aureus ATCC 6538
Candida albicans ATCC 10231
Enverionmental Isolate
5.3 Method
5.3.1 SET-1: Prepare the 100 ml of the disinfectant solution of the respective concentration as per SOP.
5.3.2 Distribute the diluted disinfectant solution in 20 ml quantities in 4 different sterile test tubes, one tube for each microorganism.
5.3.3 SET-2: Prepare another set of 4 tubes using the 20 ml of sterile purified water, one tube for each microorganism.
5.3.4 To the set 2 add the 0.1 ml of the suspension of the respective microorganism to the respective tube which will give the final concentration of 1×105 cfu/ml to 1×106 cfu/ml. Carry out the10 fold serial dilution of each microorganism up to 10-6 using 9 ml normal saline. And then plate out1 ml of each dilution. To this add 20 ml of sterile molten cooled at 45° C Soybean casein digest agar for bacterial cultures and 20 ml of sterile molten cooled at 45° C Sabouraud dextrose agar for fungi.
5.3.5 To the set 1 add the 0.1 ml of the suspension of respective Microorganism to the respective tube which will give the final concentration of 1×105 cfu/ml to 1×106 cfu/ml same as the set2. Carry out the10 fold serial dilution of each microorganism from the respective tube of set 1 up to 10-6 using 9 ml normal saline after 5 minutes, 10 minutes and after 15 minutes. And then plate out 1 ml of each dilution. To this add 20 ml of sterile molten cooled at 45°C Soybean casein digest agar for bacterial cultures and 20 ml of sterile molten cooled at 45° C Sabouraud dextrose agar for fungi.
5.3.6 Incubate the Soybean casein digest agar plates at 30-35° C for 5 days and Sabouraud dextrose agar plates at 20-25° C for 5 days.
5.3.7 After incubation observe the plates and record the count in the annexure, calculate the % reduction for each Microorganism, record the same.
5.3.8 Acceptance criteria: The respective disinfectant should show the 90 % reduction in microbial population within 15 minutes.
6.0 ABBREVIATIONS
6.1 SOP: Standard Operating Procedure
6.2 QC: Quality Control