SOP for Drain Point Monitoring in cleanrooms

1. OBJECTIVE

To lay down the Procedure for Microbiological Monitoring of Drain Points Available in Clean Rooms .

• SCOPE

This procedure is applicable for Microbiological Monitoring of Drain Points Available in Clean Rooms of Block-I Manufacturing Facility of Pharma Micro Hub Pvt Ltd

• RESPONSIBILITY
  • Microbiologist is responsible for sampling and analysis of drain points.

  • Head- Microbiology/Designee is accountable for overall compliance of this procedure

• PROCEDURE
  • Precautions:
    • Follow aseptic practices during sampling and analysis.Sanitize the hands with filtered 70% v/v IPA every time after handling the perforated drain plate or change the gloves if they are visibly dirty after handling of perforated drain plates during sampling.Perform testing as soon as possible after completion of sampling (do not store the samples).

    Preparation and Transfer of Sampling Kit.

    • Follow the procedure mentioned in Appendix-I for preparation and transfer of sampling kit to respective sampling locations.

  • Sampling Procedure.
    • Follow the respective entry and exit procedures to access the areas where the drains are located.

    • Perform monitoring of drain points mentioned below.

• Label the neutralizer swab tube with details like drain point ID, date of sampling and sign.
  • Open the perforated plate on the drain point and sample the inner surface of the drain.Unscrew the cap of the swab tube, lift the swab stick and squeeze the excess liquid by pressing the swab bud gently to the inner surface of the swab tube before sampling.Take care not to touch the bud of the swab while handling.Holding the stem, swab the sample site first with horizontal strokes and then rotate the bud and swab in vertical strokes.Swab approximately an area of 25cm² at the sample site or maximum area possible.Transfer the swabbed cotton bud back to the swab tube containing solution with neutralizer post sampling.Enter the sampling details in the format no.: MB116/F01-00 contemporaneously.Once swab sampling is completed pour sufficient quantity of water over the drain to flush out residual swab solution.Sanitize the hands between sampling of two different drains with filtered 70% v/v IPA.Transfer the swabs immediately to the MLT area for analysis
• • .Analysis of Drain Monitoring Samples.
    • Analyze the swab samples as soon as possible after receipt of samples.For testing transfer adequate number of sterile 0.1% peptone water bottles and all other accessories required for membrane filtration.Assemble the sterile membrane filtration unit under LAF. Pre-wet the membrane with approx. 10mL of sterile 0.1% peptone water.Under aseptic conditions, vortex the tube containing swab and then squeeze the swab on the inner walls of the swab tube and remove swab from the swab tube. Then, transfer 1mL of sample from the swab tube onto the membrane and filter the sample. Rinse the membrane filter with approx. 3X100 mL of 0.1% peptone water. Remove the membrane filter and place on the surface of the ready to use Soyabean casein digest agar plates (SCDA 90mm plates) supplemented with neutralizers.For negative control, repeat the steps 5.4.3 to 5.4.5 but filter 1mL of contents of one unused swab instead of sample.Incubate all the plates in inverted position at 20-25°C for 72 hrs followed by 30-35°C for further 48 hrs. However, in case of Holidays incubation can be extended up to 5 days at 20-25°C and 4 days at 30-35°C.Document the analysis and incubation details in format
• • • Observation and results reporting.Post completion of incubation, enumeration of plates shall be carried out as per the procedure mentioned in the SOP by using the colony counter .Multiply the observed counts for samples with 10 and document the results as CFU/swab in the format .
    • https://pharmamicrohub.site/how-to-validate-extended-settle-plate-exposure-time/

    Acceptance Criteria

    • NMT 100 cfu/swab for drain point samples.The negative control plate should not show growth.

  • Handling of Excursions
    • In case of any excursions obtained during microbiological monitoring of drains

    Investigation shall be performed

• Frequency of microbiological monitoring of drains.Drain monitoring shall be performed at a frequency of once in 3 months ± 7 days.Perform drain monitoring as per the schedule

  ABBREVIATIONS

• %         :          Percentage        

• • °C       :   Degree centigrade
• • cfu      :           Colony Forming Unit
• • EF      :           Effluent
• • IPA    :           Iso Propyl Alcohol
• • MB     :           Microbiology.
• • mL      :           milli liter