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 Assistant3.2 Checking: Executive/Manager
4.0 ACCOUNTABILITY
Head of the DepartmentFREQUENCY: At the change of Vendor.
5.0 PROCEDURE
5.1 Requirements :
a. Soybean casein digest mediumb. Soybean casein digest agar.
c. Sterile Petri plates.
d. Micropipette
e. Sterile micro tips
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 INOCULUM PREPARATION:
5.2.1 Inoculate 1 ml of the 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 soybean 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 a 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 a sterile test tube with the help of sterile pipette.
5.2.5 Prepare serial dilution from it & take the viable count from each dilution in duplicate.
5.2.6 Prepare suspension of above two microorganisms 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 organism)
5.5 PROCEDURE
5.5.1 Place vials in two different beakers in inverted position & then pour a sufficient quantity of 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 a plastic bag to avoid outside contamination.
5.5.5 Incubate at 30°C-35°C for 14 days.
5.5.6 Take a viable count of both suspension before and after testing & record the result in Annexure-II. Part B & C.
5.5.7 If any units show growth investigate whether it comes from the microorganism 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 microorganisms in two to three vials for each microorganism.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
| |||||
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 :
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