1.     Purpose:-                         

           To ensure use of correct method of preincubation of Medias used for routine sterility test and
           Bioburden test.

2.     Scope:-

           This procedure applies to Media’s preparation for sterility & Bioburden test performed for product
            test & plant environment monitoring.

3.     HSE Statement:-

Not Applicable.

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.
5.     Materials:-

            5.1 Flask
            5.2 Medias
            5.3 Hot plate and Stirrer

    6. Definitions:-

    6.1 Preincubation:-
                               Incubation prior to some other treatment or process

    6.2 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.
   6.3 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.Flow Chart:-

8. Description:-
8.1 Procedure:-            
8.1.1 Prepare and autoclave (if required) the required media as per SOP.
8.1.2 After successful autoclaving, take out media flasks from autoclave, tighten their screw caps and let them
 cool at room temperature.
8.1.3 Allot a lab number to all media flasks by consulting the file “Preparation & Preincubation Record”.
     Also record the same lab number of Medias in this file with all other mentioned relevant data.
8.1.4 In case of agar media, also record the total number of plates prepared with all other necessary data.
8.1.5 Place the media flasks and prepared plates in incubators at their respective optimum growth
 temperatures (e.g. Tryptic soy broth at 22.50C – 250C, Fluid thioglycollate media and agar media at 300C– 350C). Incubate broth Medias for 3 – 5 days and agar media for 24 – 48 hours.
8.1.6 Check all the Medias for any growth or turbidity on daily basis till the end of preincubation period. If
any growth or turbidity is shown in any media flask or plate remove it from incubator.
8.1.7 Use only those media flasks and plates in routine tests which qualify the preincubation period
9. Forms and Records:-

10.1 Sterilization and Preincubation record of Medias
   10.2 Log of Sterilization Cycle (Autoclave
10. References:-
             United States Pharmacopeia 35

11. Distribution:-

This SOP has to be distributed in below mentioned Departments:-

Sr. NO
Distributed to
Quality Control Department

Quality Management Department


12. Revision History:-

Date                                        Changes