Media Management and Media Preparation

1.0 Objective
To lay down a procedure for Media Management and Media Preparation.
2.0 Scope
This Standard Operating Procedure is applicable for Media Management and Media Preparation to be followed at formulation plants of abc company.
3.0 Responsibility
3.1 Officer / Executive Quality Control are responsible for Media Management and Media Preparation.
3.2 Head Quality control is responsible for the compliance of this SOP.
4.0 Abbreviations and Definitions
SOP : Standard Operating Procedure
QC : Quality Control
LAF : Laminar Air Flow
HCl : Hydrochloric acid
NaOH : Sodium hydroxide
RODAC : Replicating Organism Detection and Count
GPT : Growth Promotion Test
5.0 Procedure
5.1 Preparation of Culture Media (Using Readymade Dehydrated Media).
5.1.1 Store dehydrated media in tightly closed packs in the dark or as directed by the Manufacturer.
5.1.2 Weight a required amount of dehydrated media and add it to the flask containing purified water.
5.1.3 Make-up the required volume with purified water.
5.1.4 Distribute the media in the flasks or tubes as required. Take 50 ml each media for final

pH check after sterilization.
5.1.5 Cover the flasks and tubes with autoclavable plastic caps / cotton plugs and wrap with

butter paper / aluminum foil.
5.1.6 Media are sterilized in the autoclave at 121.4 ºC for 20 minutes at 15-lbs. pressure unless

otherwise specified. Check the pH of the media after sterilization.
5.1.7 Enter details of prepared media in the ‘Media Preparation Record’ given in Annexure-I.
5.1.8 Also make relevant entries in the autoclave sterilization Record.
5.1.9 Check visually for any growth in the form of turbidity. Discard the media if turbidity observed.
5.2 Preparation of Media Plates:
5.2.1 Rehydrated the medium and sterilize as per the media preparation instructions.
5.2.2 Cool the media and pour in sterile Petri plates (90 mm diameter) under aseptic conditions.

The Petri plates shall be marked on the bottom position with the details of the name of the

medium, load no., date of preparation, Allow to solidify and invert the Petri plates. Pre Incubate

the plates for 48hours at 300C to 350C.
5.2.3 Check visually for any growth in the form of visible growth. Discard the media if any

visible growth is observed.
5.2.4 Store the plates after pre-incubation below 25ºC.
5.3 Preparation of Slants:
5.3.1 After rehydrating the medium dissolve the agar media & makeup the required volume with purified water.
5.3.2 Dispense about 10 ml in test tubes. Plug the tubes with non-absorbent cotton and cover

with a butter paper/Aluminum foil.
5.3.3 Autoclave the media as per SOP & cool the slant on room temperature.
5.3.4 Unload the tubes from the autoclave and place them in a slanting position under LAF.
5.3.5 Ensure that the top of the slant does not touch the cotton plug and allow to solidifying.
5.3.6 Incubate for 48 hours at the 30-35ºC temperature.
5.3.7 Check visually for any contamination. Discard if necessary.
5.3.8 Mark the tubes appropriately with the name of medium, lot no., and preparation date.
5.3.9 Store the slants, after pre-incubation below 25ºC.
5.4 Preparation of RODAC(contact) plates:
5.4.1 Rehydrate the medium and autoclave the media as per SOP.
5.4.2 Cool the media at room temperature and pour in sterile RODAC plates under aseptic conditions.

The RODAC plates shall be marked on the bottom position with the details of the name of the

medium, lot no. and date of preparation.
5.5 Traceability of Media / Sterilization Load Numbers:
5.5.1 Each lot of medium prepared and sterilized is assigned a specific lot number 8 Characters as given below.
xx-yyzzm
xx indicates Serial no. of prepared media
– indicates Hyphen
yy indicates Month
zz indicates Year
m indicates the abbreviation of the media.
5.5.2 All media lots prepared / sterilized are to be serially numbered in month wise.
5.5.3 Record the name of medium and specific lot number of all media used for microbiological

testing in the Analytical Data Sheet.
5.6 Media stock maintenance:
5.6.1 Receive the media from the supplier. Check the bottles/ boxes of media for the Manufacturing

date and expiry date.
5.6.2 Incase the expiry date is within one year (except for the solutions where the expiry date

labeled itself is one year or less), reject the same and send back to the supplier.
5.6.3 Paste the label for the media bottle / box (Refer ANNEXURE-III).
5.6.4 Store dehydrated media in tightly closed packs in the dark or as directed by the manufacturer.
5.6.5 Enter the details of the receipt of the media in the Media Stock register.
5.6.6 Enter the details in such a way that incase there are 2 boxes/ bottles of media of the same

batch 2 lines are allotted for the further record of the same.
5.6.7 The next 2-bottles/ boxes with a different batch number will be entered after that

and 2 lines will be allotted for that batch.
5.6.8 Ensure that the expiry dates of the bottles are in chronological order if more than one batch is received.
5.6.9 Incase there are smaller packets in a box each individual pack shall be labeled.
5.6.10 Ensure that the media stock is used on a First in First Out or First Expiry First Out basis

and that only one pack of medium is in use at any given time.
5.6.11 On opening a new pack, enter the date of opening on the label of the bottle/ box and

in the media stock register.
5.6.12 Follow the media preparation instructions on the label of dehydrate media box.
5.6.13 For each new batch of dehydrated media received, carry out the GPT of the media

as per SOP before using for Microbiological analysis.
5.6.14 Enter the date of release on the label on the pack and in the media stock record on

successful completion of the GPT.
5.7 Retest of The Media Bottles:
5.7.1 Check that the media is having any lumps formation, if any discard the media.
5.7.2 Media containers, which fail in GPT should not be used for testing.
6.0 Forms and Records
6.1 Media Preparation Record – Annexure-1
6.2 Media stock register – Annexure-2
6.3 Label for Media Bottle – Annexure-3
7.0 Distribution
7.1 Master copy – Documentation Cell (Quality Assurance)
7.2 Control copy – Quality Control
8.0 History