1. Purpose:-
To describe the procedure for the
preparation and sterilization of different Medias used in
Microbiological testing.
2. Scope:-
This procedure is applicable for preparation & sterilization of Medias.
3. HSE
Statement:-
Face mask and gloves should be used.
4. Responsibilities:-
i)
Manager Quality Control is responsible
to ensure that procedure & formats are followed entirely as approved.
ii)
Microbiologist is responsible
to perform the test.
iii)
Lab attendant is responsible
to assist the officer microbiologist for the preparation of test materials.
5. Materials:-
5.1 Type of media to be prepared in
dehydrated form.
5.2 Fresh hot distilled water.
5.3 Washed media
bottles.
5.4 Weighing
balance, which supports the required weight for weighing.
5.5 Hot plate
with stirrer.
5.6 Butter paper
and spatula
5.7 pH meter
6.0 Definitions:
6.1 Media:
Media is the nutritive source for the
cultivation of microorganisms. The media usually prepare from protein by acid
or enzymatic digestion. Medias are usually available in dehydrated form. We
have only to mix required measured quantity in distilled water. Follow
instructions for preparation as mentioned on the label of media container.
6.2 Sterilization:
It is the process of killing or
removing microorganisms. Media is sterilized by autoclaving at
1210C for 15 minutes
at a pressure of 15 PSI.
7.0 Flow Chart:-
8.0
Description:-
8.1 Procedure:-
8.1.1
Check the physical state and expiry date of the media.
8.1.2 Weigh accurately required amount quantity of dehydrated media as
per manufacturers
Instructions on
the label in a media bottle.
8.1.3 Mix the weighed amount of dehydrated media in
equal quantity of hot distilled water as per mentioned on the label.
8.1.4 Dissolve by continuous heating and stirring
on hot plate until solution becomes completelyclear.
8.1.5 Check the pH
of media before and after sterilization according to their specifications.
8.1.6 If media for sterility test is being
prepared, dispense equal amount of 100ml in separate media flasks and mention
the type of media on the flak.
8.1.7 Screw up the corks of media bottles keeping
them a little bit loose in order to avoid the breakage of media bottle during sterilization cycle.
8.1.8 Sterilize all Medias for which supplier’s
recommendations are available at 121 C for 15 minutes.
8.1.9 Use AUTOCLAVE STERILIZATION INDICATOR TAPE
with each load that indicates proper
sterilization.
8.1.10 Immediately tight the screw caps of all media containers after
sterilization. period and allot Lab
numbers for back
traceability.
8.1.11 Incubate all Medias at their respective optimum growth
temperatures.
8.1.12 Pre incubations of all
prepared Medias are necessary for a period of 3-5 days.
8.2 Growth Promotion Test of
Medias (Positive Control):
Growth promotion test of each lot of media
prepared should be performed as per SOP.
9. Forms and Records:-
9.1 Sterilization and Preincubation record of
Medias
9.2 Log of Sterilization
Cycle
9.3 Growth Promotion test
10. References: USP 35
11 Distributions:-
This SOP has to be distributed in below mentioned
Departments:-
|
Sr. NO
|
Distributed to
|
Received
(Current)
|
Returned
(Obsolete)
|
|
1
|
Quality Control
Department
|
|
|
|
2
|
Quality Management
Department
|
|
|
12.
Revision History:-
N/A