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Learn about the microbiological analysis of Purified water and water for
injection including the method for detection of pathogens.
Water is used in verious processes in pharmaceutical
manufacturing. Microbial contamination in water may
cause contamination in pharmaceutical products. It is
necessory to analyse the Purified Water and Water for
Injection (WFI) for microbial contamination.
2.1
In
case
of
purified water & input water add 1 ml sample in duplicate
in sterile petridishes. Pour 15 ml to 20 ml of sterile
soybean casein digest agar (cooled at 45 C). Mix
thoroughly & allow solidifying the medium.
2.2 Invert & incubate the plates at 30C to 35C for 5 days.
After incubation examine the plates for total bacterial
count. Take maximum from the two plates & calculate the
no. of cfu/ml of water sample.
2.3 For detection of yeast & mold use Sabouraud
chloramphenicol agar/Sabouraud dextrose agar with
antibiotic & incubate the plates at 20C to 25C for 5 days.
After incubation examine the plates for absence of
growth.
1.1
Streak
a
portion of enriched soybean casein digest medium on the
surface of sterile MacConkey agar medium.
1.2 Incubate the plates at 30-35 C for 18-72 hours.
1.3 After incubation presence of brick red colonies on
MacConkey agar indicate the presence of Escherichia coli.
1.4 Carry out further confirmation by streaking the
colonies on the surface of levine eosine methylene blue
agar medium.
1.5 Blue black colonies with metallic sheen confirm the
presence of Escherichia coli
2.0 Detection of Salmonella:
N,N,N1,N1,tetramethyl-4-phenylenediamine
dihydrochloride on filter paper and smear with the colony;
if there is no development of a pink colour, changing to
purple, the sample meets the requirements of the test for
the absence of Pseudomonas aeruginosa.
4.0 Detection of Staphylococcus aureus: