Page 19 - Quality control of pharmaceuticals (07-PA 704) practical

B) Test For Specified Microorganisms:

B.1) Test for Escherichia coli

1. Prepare a sample using a 1:10 dilution of not less than 1 g of the material to be examined.
Use 10 ml or the quantity corresponding to 1 g or 1 ml, to inoculate a suitable amount of
soybean casein digest broth, mix, and incubate at 30 to 35C for 18-24 hours.

2. Shake the container, transfer 1 ml of soybean casein digest broth to 100 ml of
MacConkey broth, and incubate at 42-44C for 24-48 hours.

3. Subculture on a plate of MacConkey agar. Incubate at 30-35C for 18-72 hours.
4. Growth of colonies indicates the possible presence of E. coli. This is confirmed by

confirmatory tests according to SOP No. CP/ML/GR/01.
5. The material complies with the test if no colonies are present or if the confirmatory tests

are negative.

B.2) Test for Salmonella

1. Prepare a sample using a 1:10 dilution. Use the quantity corresponding to 10 g or 10
ml, to inoculate a suitable amount of soybean casein digest broth, mix, and incubate at
30 to 35C for 18-24 hours.

2. Shake the container, transfer 0.1 ml of soybean casein digest broth to 10 ml of
Rappaport Vassilliadis Salmonella enrichment broth, and incubate at 30-35C for 18-
24 hours.

3. Subculture on plates of XLD agar. Incubate at 30-35C for 18-48 hours.

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