Sop for method validation microbiology sterility testing

EFEECTIVE DATE
SUPERSEDES NUMBER
PRODUCT NAME
BATCH NUMBER

REPORT CONTENTS

S. No.	TITLE	PAGE No.
1	REPORT PRE APPROVAL
2	OBJECTIVE
3	SCOPE
4	RESPONSIBILITY
5	TRAINING RECORDS
6	OBSERVATIONS OF CULTURE SUSPENSION
7	PREPARATION FOR FLUID THIOGLYCOLATE MEDIUM
8	OBSERVATION DATA SHEET FOR FLUID THIOGLYCOLLATE MEDIUM
9	PREPARATION FOR SOYABEAN CASEIN DIGEST MEDIUM
10	OBSERVATION DATA SHEET FOR SOYABEAN CASEIN DIGEST MEDIUM
11	OBSERVATIONS OF CULTURE SUSPENSION
12	OBSERVATION OF STATIS TEST
13	ATTACHMENT
14	DEVIATION (IF ANY)
15	CONCLUSION
16	RECOMMENDATION
17	ABBREVIATIONS
18	REVISION HISTORY
19	REPORT POST APPROVAL

1.0 REPORT PRE -APPROVAL:
PREPARED BY:

DESIGNATION

NAME

SIGNATURE

DATE

OFFICER/EXECUTIVE

(QC MICROBIOLOGY)

REVIEWED BY:

DESIGNATION	NAME	SIGNATURE	DATE
EXECUTIVE/MANAGER (QC MICROBIOLOGY)
HEAD (PRODUCTION)
HEAD (QUALITY CONTROL)

APPROVED BY:

DESIGNATION

NAME

SIGNATURE

DATE

HEAD

(QUALITY ASSURANCE)

2.0 OBJECTIVE:

1. To lay down a procedure for the validation of sterility test method intended for detecting the presence of viable forms of microorganisms present in or on the Pharmacopoeia preparations.
2. To establish a procedure, which will enable to show that the product should not giving any microbial inhibitory properties.
3. To establish a documented evidence on the effectiveness of culture media used for the test.
3.0 SCOPE:
3.0 The protocol is applicable for all the products (RM/FP/PM) that has been carried out for Sterility test method.
3.1 Before release of sterility of a new product it is necessary to demonstrate that the validity of the test method used by recovery of a small number of micro-organisms in the presence of the product.
3.2 It may be performed concurrently with the actual test for sterility but should be confirmed as successful before the results of the sterility test are interpreted.
3.3 Validation is to be performed when the test for sterility has to be carried out on reformulated or new product, or whenever there is a change in the experimental conditions of the test.
3.4 All validation procedures should be carried out by personnel who are responsible for the routine testing of the product and should be done for each facility manufacturing that product.
3.5 This protocol is applicable to all the products tested for sterility by membrane filtration.

4.0 RESPONSIBILITY:

DEPARTMENTS	RESPONSIBILITIES
Quality Control	· Preparation, Review of Validation Report data. · To provide all applicable analytical procedures and documentation necessary for execution of this protocol. · Monitoring and Sampling at the different stages. · Compilation of Validation Report. · To test the sample collected and provide all Analytical Data
Production	· To review of Validation Report. · Follow up for providing the samples for completion of Validation.
Quality Assurance	· To review of Validation Report

5.0 TRAINING RECORDS:

S. No.	Name of Trainee	Department	Designation	Acceptance Criteria	Signature of Trainee	Checked by (Sign & Date)
1.				All personnel involved in execution of this protocol shall be trained in the required procedure and shall be documented
2.
3.
4.

Name of the Trainer: _______________

Inference:____________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________

Reviewed By
(Sign & Date)

6.0 OBSERVATION OF CULTURE SUSPENSION:

Name of the organism	CFU / Plate		Average
Name of the organism	Plate -1	Plate -2	Plate-1+ plate-2 2
Ps. aeruginosa
S. aureus
Clostridium sporogenes
B. subtilis
C. albicans
A. niger

Remarks:

Analyst Checked by
(Sign & Date): (Sign & Date):

7.0 PREPARATION FOR FLUID THIOGLYCOLATE MEDIUM :

7.1 Test : Sample + Fluid thioglycollate medium
7.2 Product Positive Control :
Sample + Clostridium sporogenes + Fluid thioglycollate medium ( PPC – A)
Sample + Pseudomonas aeroginosa + Fluid thioglycollate medium ( PPC – B)
Sample + S. aureus + Fluid thioglycollate medium ( PPC – C)
7.3 Positive Control :
Clostridium sporogenes + Fluid thioglycollate medium ( PC – A)
Pseudomonas aeroginosa + Fluid thioglycollate medium ( PC – B)
S. aureus + Fluid thioglycollate medium ( PC – C)

7.4 Negative Control : Fluid thioglycollate medium

Remarks :

Analyst Checked by
(Sign & Date) (Sign & Date) :

8.0 OBSERVATION DATA SHEET FOR FLUID THIOGLYCOLLATE MEDIUM

Medium : Fluid thioglycollate medium (Lot No: ________________ )
Incubation Temp : 30 to 350C (Incubator ID No. : ______________ )
Incubation Period : 14 days Date of Analysis:
Method : Date of Report:

S. No.	Date	Sample	Product Positive Control			Positive Control			Negative Control	Observed By (Sign/Date)
S. No.	Date	Sample	PPC – A	PPC -B	PPC -C	PC-A	PC-B	PC-C	Negative Control	Observed By (Sign/Date)
1
2

Analyst Checked by
(Sign & Date): (Sign & Date):

9.0 PREPARATION FOR SOYABEAN CASEIN DIGEST MEDIUM:

9.1 Test: Sample + Soyabean Casein Digest medium
9.2 Product Positive Control:
Sample + Bacillus subtilis + Soyabean Casein Digest medium ( PPC – A)
Sample + Candida albicans + Soyabean Casein Digest medium ( PPC – B)
Sample + Aspergillus niger + Soyabean Casein Digest medium ( PPC – C)
9.3 Positive Control :
Bacillus subtilis + Soyabean Casein Digest medium ( PC – A)
Candida albicans + Soyabean Casein Digest medium ( PC – B)
Aspergillus niger + Soyabean Casein Digest medium ( PC – C)

9.4 Negative Control : Soyabean Casein Digest medium

Remarks :

Analyst Checked by
(Sign & Date): (Sign & Date):

10.0 OBSERVATION DATA SHEET FOR SOYABEAN CASEIN DIGEST MEDIUM

S. No.	Date	Sample	Product Positive Control			Positive Control			Negative Control	Observed By (Sign/Date)
S. No.	Date	Sample	PPC – A	PPC -B	PPC -C	PC-A	PC-B	PC-C	Negative Control	Observed By (Sign/Date)
1
2

Analyst Checked by
(Sign & Date): (Sign & Date):

11.0 OBSERVATION OF CULTURE SUSPENSION FOR STATIS TEST:

Name of the organism	CFU / Plate		Average
Name of the organism	Plate -1	Plate -2	Plate-1+ plate-2 2
Ps. aeruginosa
S. aureus
Clostridium sporogenes
B. subtilis
C. albicans
A. niger

Remarks:

Analyst Checked by
(Sign & Date): (Sign & Date):

12.0 OBSERVATIONS OF STASIS TEST:

FTM Lot No. – SCDM Lot No. –
Incubation Temperature: Incubation Temperature:
Incubation Period: Incubation Period:
Incubator ID.: Incubator ID.:
Date of analysis: Date of analysis:
Date of observation: Date of observation
For FTM:

S. No	Date	Sample + FTM + P. aeruginosa	Sample + FTM + S. aureus	Sample + FTM + Cl. Sporogenes	Observed By/Date
1
2
3

For SCDM:

S. No	Date	Sample + SCDM + C. albicans	Sample + SCDM + A. brasilensis	Sample + SCDM + B. subtilis	Observed By/Date
1

Conclusion:

Analyst Checked by
(Sign & Date): (Sign & Date):

13.0 ATTACHMENTS:
1. Records for all critical parameters with graphical representation where applicable.
2. Raw data generated during the execution of this protocol.

14.0 DEVIATION (IF ANY):
……………………………………………………………………………………………………………………………………………………………………………………………………………………
15.0 CONCLUSION:
…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………
16.0 RECOMMENDATION:
…………………………………………………………………………………………………………

…………………………………………………………………………………………………………

17.0 ABBREVIATIONS:

Ltd. Limited
SOP Standard Operating Procedure
°C Degree Celsius
QA Quality Assurance
NA Not Applicable
FTM Fluid Thioglycollate medium
SCDM Soyabean Casein digest medium
PPC Positive Product Control
PC Positive Control
CFU Colony Forming Unit
IP Indian Pharmacopoeia
IH In-House
18.0 REVISION HISTORY:

Revision No.	Change Control No.	Detail of Changes	Reason for Change	Effective Date	Done By
00	NA	NA	New Protocol
		Incorporate observation sheet of culture suspension for statis test at point no. 11.0	Report is to be revise for up-gradation & betterment of documentation system