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SOP for Microbiological Integrity Testing of Vials


Standard operating procedure to ensure the seal integrity of vials using P.aeruginosa-ATCC 9027 and E.Coli-ATCC-8739.

1.0 OBJECTIVE

To check the seal integrity of vials.

2.0 SCOPE

To ensure the seal integrity of vials for approval of new vendors.

3.0 RESPONSIBILITY

3.1 Doing : Technical Assistant

3.2 Checking : Executive/Manager

4.0 ACCOUNTABILITY

Head of the Department
FREQUENCY : At the change of Vendor.

5.0 PROCEDURE

5.1 Requirements :

a. Soybean casein digest medium
b. Soybean casein digest agar.
c. Sterile petriplates.
d. Micropipette
e. Sterile microtips
f. Sterile 2 litre capacity beakers (2 Nos).
g. Test tube stand
h. Sterile syringe & needle
i. P. aeruginosa-ATCC 9027
j. Sterile saline (0.9% w/v)
k. E.Coli-ATCC-8739

5.2 INOCULAM PREPARATION :

5.2.1 Inoculate 1 ml of frozen culture of both organism P.aeruginosa-ATCC 9027 and E.Coli-ATCC-8739 from cryovial to 2 x 20 ml (Two tubes for each) tubes containing soyabean casein digest medium. Incubate all tubes at 30°C-35°C for 24 hrs.
5.2.2 Spread one ml of growth culture from each tube on separate plate containing SCD agar. Incubate at 30°C-35°C for 24 hrs.
5.2.3 Add 5 ml 0.9% sterile saline in one of the plates of each organism.
5.2.4 Collect the growth in sterile test tube with the help of sterile pipette.
5.2.5 Prepare serial dilution from it & take viable count from each dilution in duplicate.
5.2.6 Prepare suspension of above two micro organism to get final concentration 10^6 cfu/ml. Record the result in Annexure-II Part A.

5.3 PRE INCUBATION OF MEDIA FILLED VIAL :

Pre incubate the media filled and sealed vial at 30-35°C for about 2-3days to ensure that the vials are free from contamination.

5.4 SAMPLING QUANTITY: (20 vials for each organisms)

5.5 PROCEDURE

5.5.1 Place vials in two different beakers in inverted position & then pour a sufficient quantity of the each suspension respectively such that the closure and vial neck are covered completely.
5.5.2 Keep it for 30 min.
5.5.3 Slowly take out the vials one by one & place it on the stand in inverted position. For two different organisms, use separate stand..
5.5.4 Cover the stand of both organisms with plastic bag to avoid outside contamination.
5.5.5 Incubate at 30°C-35°C for 14 days.
5.5.6 Take viable count of both suspension before and after testing & record the result in Annexure-II. Part B & C.
5.5.7 If any units shows growth, investigate weather it comes from the micro organism used for the test or due to contamination.
5.5.8 Records the result in Annexure-I.

5.6 GROWTH PROMOTION TEST :

5.6.1 After completion of 14 days, do the growth promotion test using both micro organism in two to three vials for each micro organism.
5.6.2 Record the results in Annexure-I.

6.0 ABBREVIATIONS

6.1 °C = Degree centigrade


ANNEXURE-I
MICROBIOLOGICAL INTEGRITY TESTING OF VIALS
                                                                                      DATE  :
NAME OF M/c :_________________ M/c No :_______________________
B.No. Vial closure :__________________ Type of M/c : ___________ 

Sr.No.

Name of organisms
ATCC No:
1


2



DATE OF TESTING
TESTING
PERFORMED BY
NAME OF
ORGANISM
EXPOSURE TIME
NOTED &
CHECKED BY
REMARK



P. Aeruginosa
Start Time


End Time


E.Coli
Start Time


 End Time
After 14 days incubation.
Results :

TOTAL NO.
OF VIALS
NAME OF
ORGANISM
NO. OF VIAL 
SHOWING GROWTH.
DT. OF
OBSERVATION
IDENTIFICATION
RESULTS











Evaluation of Results  :
Done by                                              Checked by                                                   Approved by      
Date :                                                  Date :                                                               Date :

ANNEXURE-II
MICROBIOLOGY INTEGRITY TESTING OF VIALS
                                                                                                    DATE :

A)   Viable count at time of inoculum preparation :

Name of Organism
Dilution
No. of colonies
Mean
Remarks

P.Aeruginosa
















E.Coli
























B) Viable count at starting time : (Before Testing)

Name of Organism
Dilution
No. of colonies
Mean
Remarks

P.Aeruginosa
















E.Coli
























C)  Viable count after exposure time : (After Testing)

Name of Organism
Dilution
No. of colonies
Mean
Remarks

P.Aeruginosa
















E.Coli
























Evaluation of Results :
Done by                                                        Checked by                                Approved by  
Date :                                                             Date :                                           Date :         
 
Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
Email: .moc.enilediugamrahp@ofni Need Help: Ask Question


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