Procedure for Growth Promotion Test in Microbiology

1.0 OBJECTIVE:

To lay down the procedure for the Growth Promotion Test in Microbiology Laboratory.

2.0 SCOPE:

This SOP applies to carry out the procedure of Growth promotion test in the Microbiology Laboratory.

3.0 RESPONSIBILITY:

3.1 Executive – to Provide training to new joiners.

3.2 Microbiologist

3.2.1. To perform activity and preparation of SOP.

3.2.2. Keep data as GDP.

4.0 DISTRIBUTION:

Master Copy – Quality Assurance Department

Control copy- Quality Control Department

5.0 PROCEDURE:

5.1 Preparation of Inoculums for Test:

5.1.1 Take a loopful of culture from slant and inoculate in sterile 10 ml saline solutions. Mark this as a stock solution

5.1.2 Take 1.0 ml from stock solution; add in other tube containing 9 ml sterile saline solutions. Mark this as 10-1.

5.1.3 Mark serial dilution up to 10-6.

5.1.4 Mix thoroughly every dilution before transfer it to the test tube.

5.1.5 Pipette 1.0 ml of every dilution from 10-3 to 10-6 tubes into sterile Petri plates in duplicate.

5.1.6 Add approximately 15 to 20 ml of sterile SCDM medium previously cooled to 45°C into Petri plates and mix well. Leave the plate under Laminar airflow for solidification.

5.1.7 Inoculate bacterial culture plate at 32.5 ±2.5°C for two days and yeast and fungal culture at 22.5 ±2.5°C for three to five days.

5.1.8 Select the dilution giving 10-to 100 CFU/ml and use the growth promotion test of the medium.

5.2 Testing the medium for growth-promoting ability:

5.2.1 Prepare required volume of medium

5.2.2 Prepare Petri plates, tubes as per requirements.

5.2.3 Add 1.0 ml of 10 to 100 CFU/ml dilution of a specific organism to inoculums medium in duplicate, which is mentioned in Annexure–1

5.2.4 Incubate bacterial culture at 32.5 ±2.5°C for two days and yeast and fungal culture at 22.5 ±2.5°C for three to five days.

5.2.5 Observe the growth of culture; if a similar appearance matches USP, BP, and IP standards, then medium passes the growth promotion test.

5.2.6 Record the results in the format of the growth promotion test of the medium as mentioned in Annexure-II.

5.3 Perform the test on receipt of every new lot/batch of medium.

6.0 ABBREVIATION/ DEFENITION:

BP: British Pharmacopoeia

CFU: Colony Forming Unit

DGM: Deputy General Manager

IP: Indian Pharmacopoeia

LAF: Laminar Air Flow

QA: Quality Assurance

QC: Quality Control

SCDM: Soya Bean Casein Digest Medium

SOP: Standard Operating Procedure

USP: United States Pharmacopoeia

BP: British Pharmacopoeia

GDP: Good Document Practices

Growth Promotion Test annexure I
ANNEXURE II