1.0 OBJECTIVE
To lay down the procedure of sampling (before sanitization and after sanitization) and testing of drain water, in the solid dosage manufacturing area.2.0 SCOPE
This procedure is applicable to all critical drains of the Solid Dosage manufacturing area.3.0 RESPONSIBILITY
3.1 Doing: Technical Assistant/Executive.3.2 Checking: Executive/ Manager
4.0 ACCOUNTABILITY
Head of the Department5.0 PROCEDURE
5.1 Sampling Procedure
5.1.1 Collect approx 20 ml of drain sample (Before and after sanitization) in a sterile container from the drainage using a sterile pipette.5.1.2 Use sucking bulb and Pipette for the collection of the sample.
5.2 Testing Procedure for TOTAL VIABLE COUNT
5.2.1 Take 1 ml sample and dilute to 10 ml with the sterile Soyalecithin medium. (1:10 dilution )5.2.2 Take 1 ml from 1:10 dilution & further dilute to 10 ml with sterile Soya lecithin medium (1:100 dilution). Similarly prepare 1:1000 dilution.
5.2.3 Finally take 1 ml sample solution from 1: 100 dilution or 1:1000 dilution with the help of sterile micropipette into sterile Petri dish aseptically for Bacteria & fungi count.
5.2.4 Perform the test in duplicate.
5.2.5 Pour approximately 20 ml of Soyabean Casein digest agar for bacteria and Sabouraud dextrose agar for fungi, previously melted and cool to 45°C into two different plates and mix well by tilting or rotating. Allow solidifying.
5.2.6 Write down the location No. and date on each plate.
5.2.7 Incubate the SCD agar plate at 30°-35°C for bacteria and Sabouraud dextrose agar plate at 20°-25°C for fungi. for 5 days
5.2.8 After the incubation period, examine the plate for growth. (No. of colonies)
5.2.9 Calculate the average colonies of two plates in terms of cfu/ml.
5.2.10 Count the No. of colonies as per following calculation and record in Annexure –I
5.3 Calculation
5.3.1 1 : 100 à If the plate show N No. of cfu, the No. of cfu/ml = N x 1001 : 1000 à If the plate show N No. of cfu, the No. of cfu/ml = N x 1000
5.4 Testing procedure of for Microbial Limit Test (E.coli & Salmonella)
5.4.1 Take 10ml of sample and add in sterile Soyalecithin Medium and proceed as per
Method No.: ____________
5.4.2 Record the results in Annexure - II
Frequency: Once in a month.
6.0 ABBREVIATIONS:
SOP - Standard Operating Procedure
ANNEXURE -I
MICROBIAL MONITORING OF DRAINAGE IN SOLID DOSAGE AREA
S.O.P NO.: ______________
Name of Sample : Drain Water
Location: __________________________________ |
Report No
Drain ID No |
: _______________
: _______________
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Method: Plate Method
Medium Used : S.C.D Agar,
S.D Agar,
Diluents: Soya lecithin medium, Lot No. :__________
|
Incubation Condition
SCD Agar: At 30-35°C for 5 days
S.D. Agar: At 20-25°C for 5 days
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Date of Testing
|
Sample
Dilution
|
Dilution
Factor
(F)
|
cfu/plate (P)
|
Cfu/ml
F X P
|
Mean cfu/ml
| |||||||
Bacteria
|
Fungi
|
Bacteria
|
Fungi
|
Bacteria
|
Fungi
| |||||||
Before Sanitiz-ation
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After Sanitiz-ation
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Before Sanitization:
Bacteria: ______________ cfu/mlFungi : ______________ cfu/ml
E.coli : ______________
Salmonella : ______________
|
After Sanitization:
Bacteria: ______________ cfu/mlFungi : ______________ cfu/ml
E.coli : ______________
Salmonella : ______________
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Acceptance Criteria for After Sanitization : NMT 1000 cfu/ml for Bacteria
NMT 100 cfu/ml for Fungi
E.coli and Salmonella Should be Absent
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Conclusion:
Analyst : Date of Completion : Checked By :
Date: Date
| ||||||||||||
ANNEXURE -II
MICROBIAL MONITORING OF DRAINAGE IN SOLID DOSAGE AREA
S.O.P NO.: ______________________
Name of Sample : Drain Water
Location: ______________________________ |
Report No
Drain ID No
Method No.
|
: _______________
: _______________
:
| |
| TEST FOR E.coli: (a) 1st day date:______________ 10 ml of sample + 100ml Sterile Soyabean Casein Digest Medium, Lot No.:_____________________ Incubate for 24 hours.
(b) 2nd day date:_______________ 1ml of above suspension (a) + 10ml of MacConkey’s Broth,
OBSERVATION: Date:__________________ Acid & Gas Production: Yes / No
(c) 4th day date:______________ 0.1ml of the above suspension(b) inoculate in Peptone water, Lot No.__________________ and MacConkey’s Broth, Lot No: ________________Incubate for 24 hours.
OBSERVATION: Date:_________________
(1) MacConkey’s Broth: Acid & Gas Production Yes / No
(2) INDOL Test : Positive Yes / No
| |||
Conclusion: E.coli is Absent / Present
Positive Control: | |||
TEST FOR Salmonellae Spec.
(a) 1st day date:_______________10 ml of sample + 100ml Sterile Soyabean Casein Digest Medium, Lot No.:_______________________ Incubate for 24 hours.
(b) 2nd day date:_____________1ml of the above suspension (a) + 10ml of Selenite Cystine Broth,
Lot No._________________ or Tetrathionate Bile Brilliant Green Broth, Lot No.________________ Incubate for 48 hours.
(c) 4th day date:________________ Streak 1 loop full of the above suspension(b) on to the
Bismuth Sulphite Agar Plate Lot No.____________________ Incubate for 24 hours.
Brilliant Green Agar Plate, Lot No.__________________ _ Incubate for 24 hours.
Deoxycholate Citrate Agar Plate, Lot No.____________________ Incubate for 24 hours.
OBSERVATION: Date:________________
1. Bismuth Sulphite Agar: : Black or Green colony : Yes / No
2. Brilliant Green Agar : Small, Transparent, Pinkish or white colonies : Yes / No
3. Deoxycholate Citrate Agar : Colorless, Opaque with or without black centered :Yes / No
(d) 5th day date :___________________Streak & stab 1 loop full on TSI, Lot No._________ and inoculate in
Urea broth, Lot No.:_____________ with typical colonies present on above medium plates. Incubate for 24 hours.
OBSERVATION: Date:__________________
1.Slant –alkaline (Red), butt- acidic (yellow) + Gas (with or without blacking) : Yes / No
2.Red colour observed in Urea Broth : Yes / No
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Conclusion: Salmonella Spec. is Absent / Present
Positive Control: | |||
Analyst Date of Completion Checked By
Date Date
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good SOP
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