Monitoring of clean rooms- visible particle by surface swabs

1.0 OBJECTIVE : To describe the procedure for monitoring the surfaces of the aseptic processing area by surface swabbing, the media to be used for monitoring, sampling procedure, sampling locations, sampling time and frequency, incubation and interpretation of results.

2.0 SCOPE This is applicable to surface swabs sampling in aseptic processing area of production Black.

3.0 RESPONSIBILITY : Microbiologist

4.0 ACCOUNTABILITY: Head – Quality control and Head – Quality Assurance

5.0 PROCEDURE

5.1 Requirements

5.1.1 Sterile swabs

5.1.2 Sterile water for injection for dipping swabs

5.1.3 90 mm petri plates with sterile soya bean casein digest agar media containing 0.5% glycerol v/v and 4% Polysorbate 20 v/v.

5.2 Media and Plate preparation

5.2.1 Prepare the media as per SOP and additionally incorporate 4 % polysorbate 20 v/v in the media meant for plates that are to be used for monitoring surfaces.

5.2.2 Prepare the plates as per SOP.

5.3 PLATES

5.3.1 Use sterile soyabean casein digest agar plates prepared as per SOP to monitor the aseptic processing area.

5.3.2 Collect the required number of plates that are preincubated at least for 48 hours from the incubator.

5.3.3 Inspect each plate thoroughly for any colony, suspect colony, crack, condensate or dehydration in a black and white background.

5.3.4 If any such plate is observed, record in the log book and discard as per SOP no. .

5.3.1 Transfer the inspected good plates to Helmut block in a clean sanitized canister through IPQA pass box.

5.3.5 Follow the dress code and entry procedure for personnel entry to Production block and component preparation area.

5.3.6 Follow the procedure for material entry through material pass box to APA and transfer the plates to the material pass box of APA.

5.3.7 Sanitize the tray used for keeping plates in the material pass box with 0.2 m filtered 70% IPA solution and sterilized lint free pads.

5.3.8 Sanitize the plates both the side and the periphery with 0.2 m filtered 70% IPA solution and sterilized lint free pads.

5.3.9 Leave the plates in the material pass box under U.V light for 15 min. on the inverted side by spreading on the tray.

5.3.10 Expose the plates to U.V light on the other side for 15 min.

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

5.3.12 Again sanitize the plates 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. with the marker dedicated for that area.

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

5.4 SWABS

5.4.1 Wrap the swabs with parchment paper in a clean and sanitized area.

5.4.2 The day before the swab sampling, pack the swabs in a beaker and autoclave the swabs and water collected in test tubes for moistening the swabs for sampling.

5.5 SAMPLING PROCEDURE

5.5.1 Take the sterile swab, sterile water and the sanitized plate to the location where the sampling has to be done.

5.5.2 Take out the sterilized swab from the beaker and remove the parchment paper and hold the swab at one end.

5.5.3 Moisten the other end of the swab in sterile water.

5.5.4 Swab the sampling site by rolling the swab in 25cm2 area in a rolling fashion vertically and horizontally.

5.5.5 Streak the sampled swab on the petri dish containing sterile Soybean Casein Digest Agar with 0.5% glycerol v/v and 4 % polysorbate 20 v/v.

5.5.6 Similarly take the swabs at all the locations as given in the attached format Annexure I.

5.5.7 Moisten one swab with sterile water and streak directly on the agar surface to serve as Negative control.

5.5.8 Moisten one swab in a mixed culture of bacteria and fungi, and streak on the agar surface to serve as positive control. Record the no. of colonies observed.

5.5.9 Record the details of activity, name of the personnel, sampled by, time of sampling etc. in the format annexure I.

5.6 INCUBATION

5.6.1 Transfer the plates to the walk-in incubator in QC lab at 30 –35°C.

5.6.2 Incubate the plates in inverted position for 48 hrs at 30 –35°C and read the plates for bacterial colonies after completion of 48 hrs.

5.6.3 If there is a holiday then read the plates on the next working day.

5.6.4 while reading, record the results for each plate in the format – Annexure I.

5.6.5 Transfer the plates to the other walk-in incubator having 20-25°C and incubate for next 72 hrs. Read the plates for mold and yeast colonies on each day of incubation since mold may overgrow the entire surface of the plate.

5.6.6 If there is a holiday then read the plates on the next working day.

5.6.7 While reading, record the results for each plate in the format – Annexure I.

5.6.8 After completion of incubation of the plates, take the plates having counts for identification of the isolates obtained.

5.6.9 Give the reference number to each isolate as per the SOP and enter the reference number into the format – Annexure I.

5.6.10 Carry out the identification of isolates as per SOP

5.6.11 After completion of the incubation period, fill format, sign, check and submit to QA.

5.7 Sampling frequency:

5.7.1 All the locations to be monitored once in a week.

5.7.2 The critical locations for each line as given below should be monitored daily whenever there is filling activity after completion of filling and before cleaning.

Locations to be monitored daily after filling activity

Line I

1.Filling Nozzles (from outside)

2.MAR bowl bottom (stoppers)

3.MAR bowl track (stoppers)

4.Stopper chute

5.Pick up wheel (stoppers)

6.Vacuum wheel

7.Feed worm (vials)

8.Star wheels

9.Forceps

PFS line

1.Needle shield / Tip cap bowl

2.S.S insertion tube

3.Stopper bowl

4.SAG infeed track

5.SAG outfeed track

6.Pusher rod

7.Filling nozzles

8.Filling pumps

9.Pregassing nozzles

10.Needle shield / Tip cap tongs

11.Syringe support rods

12.Filling star wheels

13.Forceps used for clearing needle shield / Tip cap jams

14.Forceps used for clearing stopper jams

15.Empty magazines

AMPOULE LINE

1.Filling nozzles (First four)

2.Feed worm (next four)

3.Forceps

4.Pre and post gassing nozzles

Note: When there is filling activity the swabs should be taken at the end of the operations before cleaning and should not be taken during operations.

5.8 Interpretation of Results:

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

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

5.8.3 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.

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

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

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

5.9 Acceptance criteria for APA: