STERILITY TESTING

STERILITY TESTING 

1.0       OBJECTIVE

To lay down a procedure for STERILITY TESTING

2.0        RESPONSIBILTY

            Microbiologist/ Q.C Executive

3.0        ACCOUNTABILITY

            Quality  Control  Manager

4.0               PROCEDURE           

4.1       MEMBRANE FILTERATION METHOD

4.1.1     Enter the area as per SOP

4.1.2     Carry out environmental monitoring test as per SOP

4.1.3            Follow S.O.P  for transferring materials inside the sterile area.

4.1.4     Disinfect LAF station wth 70% filtered IPA solution & check the manometer reading (i.e 

             between 10-    

                   15 mm of water) then rinse hands with 70% filtered IPA solution followed by drying hands 

                    under the LAF before proceeding for testing.

                         For dry powder injections-Inject 5-10ml of sterile peptone water into each vial of   

                         injection(powder) by a sterile syringe after opening the flip-off seal  of the vial and dissolve 

                         powder of the vial to be tested under RLAF & suck out the sample.

For liquid injections-Suck out the sample with a sterile syringe after opening the flip-off seal  of the vials to be tested under LAF.

4.3   Collect the sample in a empty sterile flask.

4.4   Place the sterilized filtration units on the filtration assembly under the RLAF which is connected  to a a collecting flask and with a vacuum pump by a tubing tightly fitted with it.

4.5   Open the filtration units (A suitable unit consist of closed reservoir and receptacle between which a properly supported membrane of Pore size of 0.22m and a diameter of approx. 47 mm. At flow rate of 55 to 75ml of water per minute at a pressure of 70 cm of mercury)  carefully and pour the collected sample in the sterile filtration assembly and filter the sample.

4.9  Start the vacuum pump as per SOP No. K/QC/092

       4.10 Rinse the membrane filter with 3 x 100ml of peptone water.

       4.11For sterility testing for STERILE WATER FOR INJECTION or any liquid injections, rinse thoroughly  

             the outer portion of 20 nos. of ampoules/ vials and suck out water with the help of sterile syringe. 

      4.12 Collect the sample in a sterile conical flask and filter it through 0..45m membrane filter and rinse the  

             membrane filter with 100ml of 0.1%sterile peptone water and in case of liquid injections ,rinse the  

             membrane filter with 3 x 100ml of 0.1%sterile peptone water.

      4.13 After completion of filtration ,cut the membrane filter into two with the help of sterile scissor and  

              forceps and inoculate each half portion of it into sterile 100ml SCDM and FTGM

      4.14 After inoculation, close the mouth of the container tightly and incubate SCDM tubes at 20-25°C and 

             FTGM at 30-35°C.

      4.15 Hold the incubated tubes at the above mentioned temperature in incubator for duration as mentioned in  

               pharmacopoeia .

       4.16 Note the observations in the format as in annexure-I. For any adverse result inform Manager QC &         

               follow SOP no.- K/QC/087.

5.0       ABBREVIATIONS :

SOP = Standard Operating Procedure

6.0        REFERENCES

USP 25 Page no.:- 1878 , E.P: - Page no. 125  , 2.6.1

7.0        ANNEXURES : Annexure I & II

ANNEXURE- I

QUANTITIES OF THE PRODUCT TO BE EXAMINED IN THE TEST OF STERILITY (AS PER  E.P)

Type of preparation	Quantity per container	Minimum qty to be used for each medium ,unless otherwise justified & authorized
Parenteral preparations	Liquids
	Less than 1 ml	The whole content of each container
	1 ml or more	Half the content of each container but not more than 20ml
	Solids
	Less than 50 mg	The whole content of each container
	50 mg or more but less than 300 mg	Half the content of each container
	300 mg or more	150 mg

MINIMUM NUMBER OF ITEMS RECOMMENDED TO BE TESTED(AS PER  E.P)

Number of items in the batch	Minimum no. of items to be tested for each medium
Not more than100 containers	10% or 4 containers, whichever is greater
More than 100 but not more than 500 containers	10 containers
More than 500 containers	2% or 20 containers, whichever is least

ANNEXURE -II

STERILITY TEST REPORT

                                                          (MEMBRANE FILTRATION METHOD)                                 Page 01 of  02

AR. NO./GR. NO.		TEST REPORT NO.		SAMPLE  QTY.
SAMPLE
BATCH  NO.		MFG. DATE		EXP. DATE
QTY./BATCH SIZE		DATE OF SAMPLING		SAMPLED BY
DATE OF TEST		DATE OF REPORT		ANALYZED BY

MEDIA PREPARATIONS

SOYABEAN CASEIN DIGEST MEDIUM		FLUID THIOGLYCOLLATE MEDIUM
.DATE OF PREPARATION		.DATE OF PREPARATION
MEDIA BATCH NO.		MEDIA BATCH NO
MEDIA LOT NO.		MEDIA LOT NO.

TEST RESULT

NAME OF MEDIA	SOYABEAN CASEIN DIGEST MEDIUM                               INCUBATION TEMP.  20-25°C
NO. OF DAYS  ®	1	2	3	4	5	6	7	8	9	10	11	12	13	14
GROWTH OBSERVED
NAME OF MEDIA	FLUID THIOGLYCOLLATE MEDIUM                                   INCUBATION TEMP.  30-35°C
NO. OF DAYS  ®	1	2	3	4	5	6	7	8	9	10	11	12	13	14
GROWTH OBSERVED

NEGATIVE CONTROLS

NAME OF MEDIA	SOYABEAN CASEIN DIGEST MEDIUM                               INCUBATION TEMP.  20-25°C
NO. OF DAYS  ®	1	2	3	4	5	6	7	8	9	10	11	12	13	14
GROWTH OBSERVED
NAME OF MEDIA	FLUID THIOGLYCOLLATE MEDIUM                                   INCUBATION TEMP.  30-35°C
NO. OF DAYS  ®	1	2	3	4	5	6	7	8	9	10	11	12	13	14
GROWTH OBSERVED

POSITIVE CONTROLS

MEDIA  ®

SOYABEAN CASEIN DIGEST MEDIUM

FLUID THIOGLYCOLLATE MEDIUM

DAYS  ®

1

2

3

4

5

6

7

1

2

3

4

5

6

7

S. aureus

NA

-ve

+ve

+ve

+ve

+ve

+ve

+ve

C albicans/A niger

-ve

-ve

+ve

+ve

+ve

+ve

+ve

NA

C. sporogenes

NA

-ve

+ve

+ve

+ve

+ve

+ve

+ve

INFERENCE:    SAMPLE  PASSES / DOES NOT PASSES IN STERILITY TEST AS PER IP/BP/USP/IH

-ve : - No growth    and      +ve : - Growth

MICROBIOLOGIST                                                                SIGN / DATE                                                                                                                                                                                                  

MANAGER QA/QC                                                                 SIGN / DATE