Viable Particulate monitoring by MD8 Air sampler for Anaerobic organisms

1.0 OBJECTIVE                    

To monitor the viable anaerobic contamination levels  by volumetric air sampling using  MD8 air sampler  for anaerobic organism.

2.0 SCOPE

Applicable to MD8 volumetric air sampling system of Make - Sartorious.

3.0 RESPONSIBILITY

3.1 Microbiologist

4.0 ACCOUNTABILITY

4.1 Head – Quality control

4.2 Head – Quality Assurance  

5.0 PROCEDURE  

5.1 Requirements

5.1.1 MD8 air sampler

5.1.2 90mm petri plates with sterile anaerobic agar media containing 0.5% glycerol.

5.1.3 Sterile Gelatine membrane filters

5.1.4 Sterile 70% IPA solution and sterile sponge.

5.1.5 Sterilized membrane holders (sterilize the membrane holders by dry heat at 180°C for two hrs.)

5.2 Procedure

5.2.1 Prepare the media as per SOP. Prepare the plates as per SOP. Keep the plates in anaerobic jar for 48hrs and then in aerobic conditions for 48 hrs pre-incubation.

5.2.2  Transfer the inspected good plates and sanitise the plates as per SOP.

5.2.3 Remove the packs of gelatine membranes as per the required no. (each pack has 5 membranes) from the box, sanitize and expose to U.V light as above. Keep the remaining membranes in the box safely, bring it back to QC and store at the appropriate place.

5.2.4 Enter into the aseptic processing area by following proper gowning procedure. Sop.

5.2.5 Again sanitize the plates and gelatine membrane pack from the aseptic processing side of the material pass box using sterile 70%IPA solution and sterile lint free pads and mark the location nos.

5.2.6 Remove the second layer of the aluminium foil wrapped to the sterilized membrane holders.

5.2.7 Collect the plates and keep them in sterile storage area on a trolley dedicated for this purpose.

5.2.8 Collect the plates, sterile Gelatine membranes and sterile membrane holders   from the aseptic processing area side of the material pass box, and keep them on dedicated trolley in sterile storage area.

5.2.9 Take the membrane holder and the pack of gelatine membranes at the sampling location under the LAF of sterile storage trolley. 

Note : Once the pack of gelatin membrane is cut and if all the membranes from the pack of 5 are not used for sampling, identify the left over membranes and use it for positive control. Do not use for sampling at the locations in APA.

5.2.10 The air sampling at the location as given in the diagram attached Annexure I which is reviewed as per the frequency of highest non viable particulate count.

5.2.11 Collect all the plates and transfer the plates to the material pass box and arrange in the canister.

5.2.12 One plate from the lot, unexposed is streaked with Clostridium sporogenes and kept as positive control.

5.2.13 One plate from the lot of plates ,which is used for exposure ,should be labeled as –ve control and kept under incubation without any exposure.

5.2.14 Record the details of activity, name of the personnel, sampled by, time of sampling etc.

5.2.15 Plates must be incubated in an anaerobic jar for 5 days at 30 - 35°C and read for the observations, which should be immediately recorded. After completion of the incubation period, the format should be filled signed, checked, and submitted to QA.

5.3 Sampling frequency: Once in three months

5.4 Interpretation of Results:   

5.4.1 If the positive plate does not show colonies the test should be invalidated. Check the growth promotion data.

5.4.2 If count is found to be more than the specified limit, inform QA, write the corrective action report.

5.4.3 The occurrence must be investigated as per SOP and remedial action taken to prevent or minimize its recurrence.

5.4.4 Appropriate action may include examination of Production records for any indication of an incident that would explain the finding, imposing additional monitoring and / or re-sanitizing the involved area with a suitable disinfectant. 

5.4.5 It may also be necessary until acceptable conditions have been restored. 

5.4.6 The action taken should be fully documented in the standard format .

5.4.7 The technical staff to take corrective action should review a trend analysis of plate counts of both critical and non-critical areas frequently.

5.5 Acceptance Criteria

Area Action Limits (cfu per cubic meter)

Grade A (in operation) Less than 1

5.6 Calibration 

Frequency of calibration: Once in a year.

6.0 ANNEXURES 

ANNEXURE-I