Culture Media Purpose
Culture Media, To make the culture media for the development of microorganisms in the microbiological test of raw materials, In-process sample & finished products.
Culture Media Scope
This designated SOP is applicable for the preparation of Culture Media in Microbiology Laboratory at XX Pharmaceuticals Limited.
Definitions/Abbreviation:
Culture Media: The Culture media is a liquid or gel designed to support growth of microorganisms or cells or small plants. There are different type of media for growing different type of cells.
There are two major types of growth media: those used for cell culture, which use specific cell types derived from plants or animals, and microbiological culture, which are used for growing microorganisms, such as bacteria or yeast.
Responsibilities:
The roles and responsibilities are as follows:
Officer/Sr. Officer, Microbiology
To follow the instructions of the described procedure accordingly.
Asst. Manager/Manager, Microbiology
- Ensure that the procedure is kept up to date.
- Ensure right personnel from the section are trained on this specific procedure.
- Ensure the media preparation, sterilization, requisition, maintain accurate storage & proper documentation.
- To Confirm that this SOP is technically sound and reflects the required working practices as current practices.
Head of Quality Assurance
- Approval of SOP
- To ensure the overall implementation of the SOP
Procedure
Instructions
Dehydrated media are hygroscopic & are sensitive to light, heat and moisture. They are adversely affected by extreme changes in temperature e.g. hot/ cold cycling temperatures which may occur between day and night laboratory temperatures in winter season.
Condition of Media Preparation:
- Take clean & dry flask as per required volume.
- Wear appropriate Laboratory garments, gloves and mask.
- Wear safety goggles during selection of hazardous media.
Storage Condition of Dehydrated Media:
- Mention receipt date on the label when enter into laboratory.
- Store as per directions on the label; typically below 250C in a dry area, away from direct sunlight, autoclaves, drying ovens or other heat sources, Where indicated store at (2-8)0C.
- Check expiry date on the specific label, some media have suggestively shorter shelf life than others.
- Use/maintain stock in lot/batch number order. Maintain FIFO [First In First Out], FEFO [First Expiry First Out].
- Do not open a new bottle until the previous bottle has been emptied. Ensure date with label on the supplied container when it first opened.
- After intended use, ensure the container is tightly closed and store it into the designated storage area.
- Collect/procure/order the medium in an appropriate size of container and in a quantity which harmonies to normal use requirements.
- A medium in a large container which has been opened many times will deteriorate on storage. Discard the medium if the powder is not free flowing, if the colour has changed or if it appears abnormal in any way.
- Temperature of media storage room/facility/area/location should be monitored through min./max. thermometer and limit shall be followed as per the media storage requirements.
- List of media having designated storage condition and specific pH limit shall be prepared as per Annexure-II and shall be displayed near media storage facilities.
pH Check:
Check pH of the specific medium before sterilization with calibrated pH meter. If required adjust the pH with the help of 1N or 0.1N NaOH and 1N or 0.1N HCl solutions.
Culture Media Preparation:
- Select the media as per specific requirements.
- Read the instructions on the label very carefully before preparation of the specific media.
- Weigh the media according to the instructions of the manufacturer of the supplied media.
- Close the media container tightly just after weighing in order to avoid the moisture acquisition.
- Reconstitute the media with purified water and boil it appropriately until entirely dissolve.
- Distribute the reconstituted media into clean and dry flask as per requirements.
- Cap the flask using cotton plug or screw cap appropriately.
- Transfer the media flask into the detest room for use if instructed on the label as “DO NOT AUTOCLAVE” the media.
Sterilization:
- Place all prepared media into the autoclave.
- Sterilize the media at 1210C for 15 minutes.
- Wait until completion of cycle, and collect sterilized media from the autoclave when the chamber temperature reduce at 600C.
Storage of Sterilized culture Media:
- After completion of autoclaving activities, transfer all flasks containing the broth media to the test room for use.
- Store the agar media at the warming condition (500C) into autoclave until use.
- Do not keep the prepared media for more than two weeks [14 days].
- Keep the prepared agar plate at (2-8)0C into the refrigerator for not more than two weeks.
Handling of Sterilized culture Media:
- Ensure the aseptic condition during handle of sterilized media.
- Do not de-cap or expose the sterilized media outside Laminar Air Flow or Bio-Safety Cabinet.
Record Keeping:
Keep in practice to Maintain Register for Media Preparation Record, Annexure-I and Annexure-II to keep record for list of media with storage condition and pH limit.
List of Annexure: Download Here
Annexure I Register of Media Preparation
Annexure II List of Media with storage condition and pH Limit