The SOP for testing container closure integrity by microbial ingression appears to be well-structured and organized. It includes the purpose, scope, responsibility, accountability, and detailed procedure for testing. The procedure mentions the use of a biosafety cabinet, sterile hand gloves, fresh sterile pipettes, and disinfectant for handling cultures, which is necessary to maintain the sterility of the samples and prevent contamination.
The preparation of the culture suspension and testing procedure is also clearly defined. The use of positive control and serial dilutions for culture suspension preparation ensures the accuracy and reliability of the test results. The SOP also provides information on how to record and report the results, as well as the abbreviations used in the document.
However, it is recommended to include more details on the specific equipment and materials required for testing, such as the leak test apparatus and disinfectants used, as well as the acceptance criteria for passing the test. Additionally, the SOP should mention the regulatory requirements and guidelines that the testing procedure is based on.
Purpose: The purpose of this document is to establish a procedure for testing container closure integrity by microbial ingression ( CCIT).
Scope: This procedure is applicable to the Microbiology Department at XXX.
Responsibility: The Officer/Executive in the Microbiology Department is responsible for ensuring the procedure is followed.
Accountability: The Head of Microbiology is accountable for ensuring that the procedure is properly followed and adhered to.
Procedure:
5.1 General Instructions: To ensure safety during the handling of microbial cultures, the following instructions must be adhered to:
5.1.1 Handling of culture shall always be carried out under Biosafety cabinet. 5.1.2 Use mask & sterile hand gloves while handling the culture. 5.1.3 Use fresh sterile pipette for each transfer. 5.1.4 In case of spillage of microbial cultures, immediately wash with 70% IPA disinfectant. 5.1.5 Mop the area with the 70% IPA disinfectant after completion of activity.
5.2 Preparation Culture Suspension The preparation of the culture suspension should be done as follows:
5.2.1 Take out the E. coli working cultures slants (weekly) from the refrigerator 30 minutes prior to testing, acclimatize with the working environment and transfer to the Biosafety Cabinet in the Culture handling room.
5.2.2 Spread the E. coli culture on pre-incubated 90 mm SCDA plate in duplicate on the whole surface.
5.2.3 Incubate the plates at 30-35°C for not more than 72 hours.
5.2.4 Prepare the culture suspension to achieve the 104-105 CFU.
5.2.5 Take the full culture of the plates into the sterile 100 ml sterile BSCP and vortex to mix uniformly.
5.2.6 Label this dilution as 10-1. Take 10 ml from dilution 10-1 to the next dilution of 90 ml BSCP and label as 10-2 and carry out serial dilutions similarly up to 10-6 in duplicate.
5.2.7 Transfer 1.0 ml of the inoculums from each dilution in two sterile plates.
5.2.8 Incubate the plates at 30-35°C for not more than 72 hours. 5.2.9 Until the result of culture suspension store each dilution at 2-8°C. 5.2.10 Record the results as per the current version of the Format No. XXX
5.3 Testing Procedure The testing procedure should be done as follows:
5.3.1 Visually inspect all vials for any sealing defect.
5.3.2 Send the sample to the Microbiology lab for analysis.
5.3.3 In case of media fill take 50 vials as per the respective media fill protocol.
5.3.4 Transfer each vial into the culture handling room through the dynamic Passbox.
5.3.5 Enter the MLT testing area as per the current version of the SOP No. xxx “Entry and Exit Procedure for Testing Area (MLT Room, Culture Handling Room, Plate Preparation Room)”.
5.3.6 Operate the Biosafety cabinet as per the current version of the respective SOP No.xxx “Cleaning, Operation and Maintenance of Biosafety Cabinet”.
5.3.7 Pour approximately 2 liters of culture suspension of organism E. coli having 104-105 CFU/ml into the leak test apparatus.
- Pour the approx. 2 liter of culture suspension of organism E. coli having 104-105 CFU/ml in to the leak test appartus.
- Inoculate the sufficient vial in culture suspension.
- For Positive Control:-
- Inoculation 1 vial after punching rubber stopper with needle along with the test. After completion of the operation, disinfectants the punching vial. Incubate the vial 30-35º for NMT 72 hrs.
- Operate the leak test apparatus as per current version of the SOP No. xxx/SOP/MB/048.
- Apply the vacuum of not less than -400 inch of Hg and hold for 5 minutes.
- After completion of the operation of leak test appartus collect the vials from the culture suspension.
- Disinfection the each vials with the help of schedule disinfectant and keep it for dry the vials in biosafety cabinet.
- Transfer the vials into the incubation room through the dynamic pass box as per current version of the respective SOP No. XXX/SOP/MB/037 “Cleaning and Operation of Dynamic Pass Box”.Incubate the media fill vials at 20-25 °C for NLT 7 days and 30-35 °C NLT 7 days.
- Record the result as per current version of the respective sterility test format no. xxxx
The following are the reference guidelines for testing container closure integrity by microbial ingress:
- United States Pharmacopeia (USP) General Chapter <1207> Package Integrity Evaluation – Sterile Products
- European Pharmacopoeia (Ph. Eur.) Chapter 3.2.9 Integrity of Non-Aqueous Container Closure Systems and Chapter 3.2.8 Integrity of Aqueous Container Closure Systems
- Parenteral Drug Association (PDA) Technical Report No. 27 (Revised 2017) – Pharmaceutical Package Integrity
- International Organization for Standardization (ISO) 11607-1 Packaging for terminally sterilized medical devices – Part 1: Requirements for materials, sterile barrier systems and packaging systems
- ASTM F2338 – 09 (2014) Standard Test Method for Nondestructive Detection of Leaks in Packages by Vacuum Decay Method.