Monitoring of clean rooms - Viable particles by volumetric air sampling (HI-AIR PETRI) For anaerobic organisms

1.0 OBJECTIVE

To monitor the viable anaerobic contamination levels by volumetric air sampling in aseptic processing areas.

2.0 SCOPE

2.1 Applicable to Hi air petri volumetric air sampling system of make Hi media.

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 Instrument Hi air Petri of Hi media

5.1.2 Stand

5.1.3 Cone for 90mm petri plate

5.1.4 Feeder cone circular clamp assembly for air passage

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

5.2 Prepare the media as per SOP. Prepare the plates as per SOP.

Keep the plates in anaerobic jar for 48 hrs and then in aerobic conditions for 48 hr preincubation.

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

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

5.5 Check the calibration label of air sampler and ensure that the instrument is within in the calibration period.

5.5 Collect the Autoclave sterilized SS cone and SS Feeder cone circular clamp assembly.

5.6 For operating the air sampler follow the SOP.

5.7 Perform the air sampling at the location as given in the diagram attached Annexure II which is reviewed as per the frequency of highest non viable particulate count.

5.8 After completion of the sampling, aseptically open the SS feeder cone circular clamp, and remove the petri plate . Put the lid to the petri plate that is kept in inverted position.

5.9 Collect all the plates and arrange in the canister.

5.10 One plate from the lot, unexposed is streaked with Clostridium sporogenes and kept as + ve control.

5.11 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.

5.12 Record the details of activity, sampled by, time of sampling etc.

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

5.14 Sampling frequency: Once in three months

5.15 Interpretation of Results:

5.15.1 If the positive plate does not show colonies the test will should be not be invalidated, check the growth promotion data.

5.15.2 If count is found to be more than the specified limit, the occurrence must be investigated as per SOP and remedial action taken to prevent or minimize its recurrence.

5.15.3 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.

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

5.15.5 The action taken should be fully documented.

5.16 Acceptance Criteria

Area (in operation and at rest)	Alert Limit	Action Limit
Grade A (in operation and at rest)	NA	Less than 1
Grade B (at rest)	NMT 2	NMT 5
Grade B (in operation) & and Grade C ( at rest)	NMT 5	NMT 10
Grade C (in operation) and Change Room	NMT 5	NMT 10