VALIDATION OF SELF LIFE OF IPA 70%

1. PURPOSE:

This Sop Describes the Procedure for Validation of shelf life of isopropyl alcohol 70%.

• SCOPE:

This Sop is Applicable to the Procedure for Validation of self life of IPA 70%.

• RESPONSIBILITY:

         Microbiologist

• ACCOUNTABILITY:

Head Quality Control

• PROCEDURE
  • Method
  • In order to efficiently conduct validation of diluted disinfectant storage condition and period, ensure that the following requirements are fulfilled –
  • Validated Laminar air flow
  • Calibrated Incubators
  • Soybean casein digest agar
  • Approved Test Methods for Swab testing and Plate Exposure
  • Test Organisms – E. coli, Salmonella abony, Staph. aureus, C. albicans, and Wild culture – WC1
  • Before proceeding for Validation of Shelf Life for diluted disinfectant solution, following parameters are checked –
  • Each lot of dehydrated media used for validation, the medium must be tested for its growth promoting qualities
  • VALIDATION TEST
  • Validation of diluted disinfectant storage conditions are done by following methods
  • Surface swab test Method
  • Settle Plate Method
  • Prepare 70% v/v solution of Isopropyl Alcohol.
  • Carry out the sterility of diluted IPA solution by Membrane filtration method.
  • Store the disinfectant solution at room temperature, and analyze the sample on the day of preparation, after 24 hrs, 48 hrs, 72 hrs and 96 hours by Surface swab Test, Settle Plate Method
  • SURFACE SWAB METHOD
  • Remove following culture slant from the refrigerator and allow it to attain room temperature.
  • Staphylococcus aureus
  • Escherichia coli
  • Pseudomonas aeruginosa
  • Salmonella, and
  • Wild culture
  • Inoculate loop full of the culture from each slant separately into 50 ml of sterile Soyabean casein digest medium and incubate at 32.5 ± 2.5°C for 24 – 48 hrs.
  • Transfer 1.0 ml of the broth culture into 9.0 ml of sterile saline solution (0.9% sodium chloride solution) to obtain a test dilution of 10^-1.
  • Transfer 1.0 ml of the 10^-1 dilution into 9.0 ml of sterile saline solution to give 10^-2 dilution.
  • Similarly serially dilute the culture suspension to obtain dilution of 10^-3 , 10^-4, 10^-5  and 10^-6
  • Plate 1.0 ml of the culture suspension from dilution 10^-3, 10^-4, 10^-5 and 10^-6 in duplicate into sterile petri dishes.
  • Pour approximately 15-20 ml of sterile Soybean Casein Digest Agar cooled to about 45°C in each plate. Incubate at 32.5 ± 2.5°C for 24 to 48 hours.
  • Count the number of colonies on each plate and Select the dilution, which gives a count of not less than 10^-5.
  • Apply 1.0 ml of each culture suspension containing cell concentration not less than 10^-5 cfu/ml, separately on the floor approx 25 cm2 area, and allow to air dry.
  • After drying, take a surface swab as per latest SOP for Swab Testing, and carry out the determination of total aerobic count per cm2 within 4 hours of sampling.
  • Immediately clean the floor with IPA 70% v/v solution and allow to stand for 30 minutes to facilitate the action of disinfectant solution on the challenge test organisms.
  • After 30 minutes, take a swab and detect the bacterial count as per SOP for swab testing.
  • Repeat the same procedure, step no. 9 to 12 to determine the efficacy of 70% IPA solution after 24 hrs, 48 hrs, 72 hrs and 96 hours at room temperature
  • SETTLE PLATE METHOD
  • Expose the pre-incubated sterile Soybean casein digest agar plate for 2 hours.
  • After plate exposure time, immediately clean the floor with IPA 70% v/v solution.
  • After proper cleaning, spray the area with IPA 70 % v/v solution and immediately close the room 30 minutes to facilitate the action of disinfectant solution.
  • After 30 minutes of contact time, immediately expose the plate as per latest SOP for plate exposure and detect the bacterial count per location per 2 hours.
  • Repeat the same procedure, step no. 1 to 4 to determine the efficacy of IPA 70% v/v solution after 24 hrs, 48 hrs, 72 hrs and 96 hours at room temperature.
  • ACCEPTANCE CRITERIA
  • Diluted disinfectant solution, which is stored at room temperature, is effective when the test result of surface swab and Settle plate shows 90% reduction of the challenged microorganisms.

• TRAINING:

Trainer   : Manager – Quality Control

Trainees : Staff of the microbiology departments

• DISTRIBUTION:

Master Copy                  :           QA Department

Controlled Copy            :         Microbiology Department

Display Copy                 :           Microbiology Department (If Required)

• ATTACHMENTS:

Sr. No.	Title	Annexure No.
01.	Protocol of Validation of self life of IPA 70%.	NA

• REFERENCES:

In-House

1. ABBREVIATIONS:

Abbreviations	Extended Form
SOP	Standard Operating Procedure
NA	Not Applicable
MB	Microbiology
Sr. No.	Serial Number
QA	Quality Assurance
DEPT.	Department
IPA	Isopropyl Alcohol
%	Percentage

1. REVISION HISTORY OF CHANGE:

Sr. No.	Date	Revision Details	Revision No.
01