OASIS LABORATORIES PRIVATE LIMITED, DEHRADUN

SOP No.:
QUALITY CONTROL DEPARTMENT Page 1 of 4
QCG0001
Supersedes : Effective:
STANDARD OPERATING PROCEDURE
00 April 2010
MICROBIL TESTING PROCEDURE FOR
Version No. : Review date:
TOTAL BACTERIAL, YEAST AND MOLD
01 March 2013
COUNT

1.0 OBJECTIVE: To describe the procedure for microbial testing for total
bacterial, yeast and mold counts in raw materials and
finished products.

2.0 SCOPE : Bacteria, Yeast and mold count.

3.0 RESPONSIBILITY : Q.A. / Q.C INCHARGE OFFICER

4.0 PROVEDURE :

Total Bacterial Count

Reagent : Buffer solution or 0.1% peptone solution.

Dissolve 3.56 gm of potassium dihydrogen orthophosphate 4.30 gm sodium chloride,
7.23 gm of sodium orthophosphate and 1.0 gm peptone in 1.0 liter distilled water. Or
1.0 gm peptone in 1.0 liter distilled water. Adjust pH 7.0 of above solution by o.1 N
NaOH or dulute orthophosphoric acid as required.

Distribute 90 ml of buffer solution or 0.1% peptone buffer in 2.50 ml conical flask or
150 ml test –tube. Plug conical flask and test –tube with nonabsorbent cotton plug.

Autoclave all flask and test tubes at 15 Ib. pressure for 20 minutes.

Soyabaean casein digest agar preparation. Prepare soyabean casein digest agar as
per standard operating procedure.

Cool agar at 40° C in a water bath before use for pour plates.

Method : Transfer aseptically 10 gm/ml sample as per requirement in to a 90 ml
Sterile buffer / 0.1% peptone solution containing flask.
Designation Signature Date
Prepared by Chemist - QC
Reviewed by Manager - QC
Approved by Manager - QA

30 of sodium chloride.0 lit.0 ml solution from 1:10 and 1:100 dilution to the sterile petridishes in duplicate. Above dilution is 1:10.QC Reviewed by Manager .e.1:10 to other 90 ml of sterile buffer /0. 7. Dissolve 3. If no growth is observed for two dilution’s i. Adjust pH 7. express the results not more then 10 bacteria per 10 gm or 10 ml of sample. OASIS LABORATORIES PRIVATE LIMITED.: QUALITY CONTROL DEPARTMENT Page 2 of 4 QCG0001 Supersedes : Effective: STANDARD OPERATING PROCEDURE 00 April 2010 MICROBIL TESTING PROCEDURE FOR Version No.0 gm peptone in 1. Distilled water.QC Approved by Manager . DEHRADUN SOP No.1 N NaOH or dilute orthoposphoric acid as required.1 peptone solution containing flask. Pour 20 ml of sterile agar in above mark petridish when temperature of sterile agar is 40° C aseptically. : Review date: TOTAL BACTERIAL.1 % peptone solution.1:10 & 1:100.0 liter distilled water.56 gm of potassium dilrydrogen orthophosphate 4. Invert the petridishes and incubator at 30° to 35° C for 48 to 72 hours in incubator. Or 1. Transfer aseptically 10 ml of above dilution i.0 gm peptone in 1.0 of above solution by 0. examine the plates for growth.QA . Designation Signature Date Prepared by Chemist . YEAST AND MOLD 01 March 2013 COUNT Mix will to dissolved or suspend the sample. Mix and allow the media to solidify at room temperature. Mark with marker pen on petridish for respective dilute transfer aseptically 1. After incubation period.23 gm of sodium orthophosphate & 1. Total yeast and mold count Reagent : Buffer solution or 0. Note : If count is higher then 300 colonies per plates dilute the sample further to 1:1000 or more. Count the number of bacterial colonies in all the plates. Express the average of two plates in terms of the number of bacteria per gm/ ml of sample.e.

Method: Transfer aseptically 10 gm sample as per requirement in to a 90 ml sterile buffer /0. Transfer aseptically 1 mi solution from 1:10 & 1:100 dilution to the sterile petridishes in duplicate. Plug conical flask & test-tube with nonabsorbent cotton plug.50 ml conical flask or 1. : Review date: TOTAL BACTERIAL. Mix well to dissolve or suspend the sample.QA . Pour 20 ml of sterile agar in above mark petridish when temperature of sterile agar in 40° to 45°C aseptically.1 % peptone buffer in 2.: QUALITY CONTROL DEPARTMENT Page 3 of 4 QCG0001 Supersedes : Effective: STANDARD OPERATING PROCEDURE 00 April 2010 MICROBIL TESTING PROCEDURE FOR Version No. After incubation period examine the plates for molds & yeast growth. Autoclave all flask & test-tube at 15 Ib pressure for 20 minutes.QC Reviewed by Manager . OASIS LABORATORIES PRIVATE LIMITED. Sabouraud Dextrose Agar Preparation: Prepare sabouraud dextrose agar as per standard operating procedure. YEAST AND MOLD 01 March 2013 COUNT Distribute 90 ml of buffer solution or 0. Cool agar at 40° .e. Invert the petridishes & incubate at 30° to 35° C for 48 to 72 hours in incubator. Designation Signature Date Prepared by Chemist .45° c in a water bath before use for pour plates. Mix & allow the media to solidify at room temperature for 48 to 72 hours in incubator. 1:10 to other 90 ml of sterile buffer 0. Mix well.50 ml test-tube.1 % peptone solution containing flask.QC Approved by Manager . dilution is 1:100 Mark with marker pen on petridish for respective dilutions. Above dilution is 1:100 Transfer aseptically 10 ml of above dilution i.1% peptone solution containing flask. DEHRADUN SOP No.

Designation Signature Date Prepared by Chemist . If no growth observed for two dilutions i.QC Approved by Manager . YEAST AND MOLD 01 March 2013 COUNT Count the number of mokis & yeast colony in all the plates. 1:10 &1:100 express the results not more then 10 molds & yeast per 10 gm of sample Note : If count is higher then 300 colonies per plates dilute the sample feather to 1:1000 or more.QC Reviewed by Manager . OASIS LABORATORIES PRIVATE LIMITED. : Review date: TOTAL BACTERIAL. Express the average of two plates in terms of the number of molds & yeast per gm of sample.: QUALITY CONTROL DEPARTMENT Page 4 of 4 QCG0001 Supersedes : Effective: STANDARD OPERATING PROCEDURE 00 April 2010 MICROBIL TESTING PROCEDURE FOR Version No.e. DEHRADUN SOP No.QA .