Preparation and Sterilization of Microbial Media STANDARD OPERATING PROCEDURE

	STANDARD OPERATING PROCEDURE
	Department: Quality Control	SOP No.: QC/MB/xxx
Preparation and Sterilization of Microbial Media		Effective Date:
		Review Date:
		Page No.:

1.0            OBJECTIVE:

1.1                 To lay down a procedure for the preparation and sterilization of microbiological media            using Pre - formulated / dehydrated media.

2.0            SCOPE:

2.1                 This SOP is applicable to all the media used for Microbiological testing in  Pharma Private Limited.

3.0            RESPONSIBILITY:  

3.1                   Microbiologist: Responsible for preparation of media.

3.2                   Section In charge: To review the procedure.

3.3                   QC-Head: Responsible for implementing the procedure.

3.4                   QA Head: Overall compliance

4.0            PROCEDURE:

4.1                 Receipt of Microbial media:

4.1.1             All microbial media shall be procured from any one of the reputed media manufacturer mainly from the following sources.

4.1.1.1       Hi-Media laboratory

4.1.1.2       Difco Labs

4.1.1.3       Merck

4.1.2             All media shall be received in the form of dehydrated media or ready prepared media.

4.1.3             On receipt of the microbial media ensure that the Certificate of Analysis is received along with the each media and verify the media details like lot number, Manufacturing date, Expiry date in COA with that the label information on media container.

4.1.4             If media details are matching then receive the media, if media details are not matching reject the media containers and return to supplier.

4.1.5             Enter the received media in annexure I (Media inward and stock record).

4.1.6             Allot the In-house ID number for all received dehydrated media containers.

4.1.7             Media in-house ID shall contain the details like “X-DM/YYY/ZZ”.

Where X stands for Media code & DM stands for Dehydrated Media, YYY stands for serial number shall start from 001 and ZZ stands for last two digits of current year.

For example media in-house ID for Soyabean Casein Digest Agar will be

SCDA-DM/001/17

Where SCDA is media code & DM is Dehydrated Media,

            001 is serial number and

            17 is the last two digits of current year.

4.1.8             Refer Table I in annexure IV for Media Codes of all media.

4.1.9             Label the each container with the label mentioned in Annexure III.

4.1.10         Store all the received media as per manufacturer instructions.

4.1.11         Before using the media for regular analysis perform Growth Promotion Test as per current version of SOP QC/MB/024 for each lot.

4.2                 Preparation of Media:

4.2.1             Follow the manufacturer instructions for preparation of all media.

4.2.2             Whenever the media taken for growth promotion test or for regular analysis enter detail of container ID number in Annexure I.

4.2.3             Weigh the required quantity of the dehydrated media in to a cleaned and dried suitable container which is labelled with media name and date of preparation.

4.2.4             Add adequate quantity of freshly collected purified water.

4.2.5             Stir / mix properly to get uniform dispersion of media with water.

4.2.6             Boil the prepared media on Hot plate to dissolve the media completely.

4.2.7             The pH can be checked as per label claim of Manufacturer.

4.2.8             Check the pH using a pH meter before and after sterilization. Adjust the pH, as required, by using 1M/0.1 M HCl or 1M/0.1M NaOH before sterilization as that after sterilization pH shall come within required pH, if pH after sterilization is not within the limit discard that media and prepare freshly for analysis.

4.2.9             Add supplements for specific media as per table I in annexure IV.

4.2.10         Media shall be prepared on daily basis based on the samples to be analyzed and prepared media shall be used on the same day.

4.2.11         Allot the in-house ID number for all prepared media.

4.2.12         Media in-house ID shall contain the details like “X-PM/YYY/ZZ”.

Where X stands for Media code & PM stands for Prepared Media, Y stands for serial number shall start from 001 and ZZ stands for last two digits of current year.

For example media In-house ID for Soyabean Casein Digest Agar will be   

SCDA-PM/001/17     

              Where… SCDA is media code 

                              PM is prepared medium,

                              001 is serial number and  

                             17 is last two digits of current year.

              If the same day SCDA is prepared in second autoclave load the number will SCDA-   PM/002/17.

4.2.13         Refer Table I in annexure IV for Media Codes of all media.

4.2.14         Dispense the prepared media into individual containers [flasks, tubes] as required.

4.2.15         Close the containers using cotton plugs and cover the cotton plug with aluminium foil.

4.2.16         Label the media flasks with media name and date of preparation.

4.2.17         Sterilize the media in a steam sterilizer Horizontal Autoclave refer SOP QC/MB/xxx at 121°C for 15 min, or as directed in annexure IV Table II.

4.2.18         Dispense the sterile media as required into sterile containers (Petri plates and tube) at approximately 40°C and allow to solidify.

4.2.19         Label each petri plate and test tube with In-house ID of the medium & date of preparation and use them for regular analysis.

4.2.20         For prepared solid media (each autoclaved lot), growth promotion test shall be performed by means of streaking with all organisms mentioned in current version of SOP QC/MB/xxx by rotating weekly with one organism.

4.2.21         For prepared liquid media (each autoclaved lot), growth promotion test shall be performed by means of observing turbidity with all organisms mentioned in current version of SOP QC/MB/xxx by rotating weekly with one organism.

4.2.22         The left over media after completion of analysis shall be dispose as per current version of SOP QC/MB/035.

4.3                 Preparation of 1M/ 0.1M HCl and 1M/0.1M NaOH

4.3.1             1M HCl: Dilute 85 mL of conc. HCl diluted to 1000 mL with purified water to obtain 1M HCl.

4.3.2             0.1M HCl: Dilute 10 mL of 1M HCl to 100 mL to obtain 0.1M HCl solution.

4.3.3             1M NaOH: Dissolve 40 g NaOH in purified water and make to 1000 mL to obtain 1M NaOH.

4.3.4             0.1M NaOH: Dilute 10 mL of 1M NaOH solution to 100 mL to obtain 0.1MNaOH solution.

4.4                 Precautions during media preparation and usage:

4.4.1             Label the container with opened-on date when opened for the growth promotion test / for regular usage.

4.4.2             Close the container tightly immediately after use.

4.4.3             Clean the spillage, if any, immediately sanitize the surface with 70% IPA.

4.4.4             Use hand gloves and nose masks while handling the dehydrated media.

4.4.5             Use cleaned and dried glassware for media preparation.

4.4.6             Care should be taken do not overheat the media, avoid re-melting the prepared media.

4.4.7             Avoid excessive shaking of the prepared media to minimize froth/air bubble.

4.4.8             Before use, check pre-incubated agar plates/tubes for un- even spread / drying or any contamination.

4.4.9             All prepared media must be sterilized within 4 hrs.

4.4.10         Any spillage / breakage in the sterilizer must be cleaned immediately.

4.5                 Media Reconciliation procedure:

4.5.1             The fill weight of dehydrated media will differ up to ± 2 % from the stated weight on the bottle.

4.5.2             Daily media consumption shall be entered in annexure II.

4.5.3             After completion of each dehydrated media container check for the less or excess quantity by comparing the quantity mentioned in consumption record.

4.5.4             Document the excess or less quantity which differs from consumption record in Annexure II.

4.5.5             If excess quantity will observe that quantity shall be added to the next media bottle and the same shall be documented in consumption record in Annexure II.

4.5.6             If less quantity will observe take sufficient quantity of media from next bottle to complete the media preparation and the same shall be documented in consumption record in Annexure II.

4.5.7             After completion of each dehydrated media container, reconcile in the stock record (Annexure – I).

5.0           ABBREVIATIONS:

QC: Quality Control	QA: Quality Assurance
HCl: Hydrochloric Acid	IPA: Iso Propyl Alcohol.
NaOH: Sodium Hydroxide	COA: Certificate of Analysis
SOP: Standard Operating Procedure

7.0           REFERENCES:

7.1                 United States Pharmacopeia Chapter <61> &<62> (Microbiological Examination of non-sterile Products: Microbial Enumeration test and Test for specified microorganisms).

7.2                 European Pharmacopeia chapter 2.6.12 & 2.6.13 (Microbiological Examination of non-sterile Products: Microbial Enumeration test and Test for specified microorganisms).

7.3                 United States Pharmacopeia Chapter <1117> Best Microbiology Lab Practices.