SOP for Environmental Monitoring in Production, R&D and Microbiology Laboratory

OBJECTIVE

To lay down a procedure for Environmental Monitoring in Production, sampling booths, dispensing booths, R&D, and Microbiology Laboratory.

SCOPE

This procedure is applicable to the specified areas in Production, Sampling booths, Dispensing booths, R&D, and Microbiology laboratory at XYZ Limited.

RESPONSIBILITY

Microbiologist - Execution of activity

Section In-charge – To review the activity

QC Head - To implement the procedure

QA Head - Overall Compliance.

PROCEDURE

Media Plates Preparation

• Prepare SCDA and SDA plates as per the current version of SOP and pre-incubate the prepared SCDA plates at 30°C to 35°C for 24 to 48 hours and SDA plates at 20°C to 25°C for 24 to 48 hours.
• After completion of the incubation period observe the plates, if any contamination is observed discard the contaminated plates and use the plates that are free from contamination.
• Before going to start the environmental monitoring ensure the cleaning of the respective areas.

Environmental Monitoring Procedure:

• Environmental Monitoring can be done by two methods

1. Passive air sampling method (Settle plate method) and
2. Active air sampling method (Air sampler method).

Passive air sampling method (Settle Plate Method):

• Use pre-incubated plates of soybean Casein Digest Agar (SCDA) for TAMC and Sabouraud Dextrose Agar (SDA) for TYMC.
• Label each pre-incubated SCDA and SDA plate with the name of the media, the name of the location, and the date of exposure.
• Carry pre-labelled plates in a stainless steel petri plate carrier which is previously wiped with 70% IPA to the respective locations.
• Follow the gowning procedure as required by the respective department to avoid additional contamination.
• Wipe the hands with 70% IPA and expose the plates by opening the lid on the designated plate exposure stands wherever applicable and it should be performed as per the current layout given by QA.
• Expose SCDA & SDA plates for four hours in locations mentioned in Annexure - I & II except LAF chambers in the microbiology lab, RLAF unit in sampling & dispensing rooms and pass boxes.
• Expose SCDA & SDA plates for one hour in LAF chambers in microbiology, pass boxes, and RLAF units in sampling and dispensing rooms.
• After exposure, close the lid & collect all exposed plates in a stainless steel petri plate carrier which is previously wiped with 70% IPA and bring them to the microbiology lab for incubation.
• Incubate SCDA plates at 30°C to 35°C for 72 hrs and SDA plates at 20°C to 25°C for 5 days.
• Incubate a negative control plate that is not exposed.
• Count the colony-forming units in the plate at the end of incubation time and record the results in Annexure I & II.
• Frequency: Once in a month ± 5 working days for locations mentioned in Annexure – II and daily for locations mentioned in Annexure - I.
• Limits:

1. The limit for plate exposure of Grade A locations is <1 cfu/plate.
2. The limit for plate exposure in Grade C locations is 50 cfu/plate.
3. The standard Limit for plate exposure in Grade D locations is 100 cfu/plate.
4. The alert limit for plate exposure in Grade D locations is 50 cfu/plate.
5. The action limit for plate exposure of Grade D locations is 60 cfu/plate.

• Acceptance Criteria:

1. Bacterial colonies shall be within the limit.
2. Fungal colonies shall be absent.

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Active Air Sampling method (Air Sampler method):

• Use pre-incubated plates of Soya bean Casein Digest Agar (SCDA) for the air sampling method.
• Label each pre-incubated SCDA plate with the name of the media, the name of the location, and the date of sampling.
• Carry pre-labeled plates in stainless steel petri plate carrier which is previously wiped with 70% IPA and air sampler unit to the respective locations.
• Follow the gowning procedure as required by the respective department to avoid additional contamination.
• Wipe the hands with 70% IPA, open the lid, and place the media plates in the stainless steel petri plate holder cup of air sampler carefully and perform the air sampling as per the current version of SOP No: QC/MB/036.
• Sample 1000 liters of air in 10 minutes (with the speed of 100/min) per location mentioned in Annexures - III & IV and it should be performed as per the current layout given by QA.
• After completion of air sampling, collect the plates in a stainless steel petri plate carrier which was previously wiped with 70% IPA, and bring them to the microbiology lab for incubation.
• Incubate SCDA plates at 30°C to 35°C for 48 hrs in the bacteriological incubator and then transfer to BOD incubator and incubate at 20°C to 25°C for 72 hrs.
• Incubate a negative control plate that is not exposed.
• Count the colony forming units in the plate at the end of incubation time and use the conversion table given in annexure-I of SOP QC/MB/036 (Operation and calibration procedure of Centrifugal Air Sampler) to report the results as cfu/1000 liters of air (cfu/m3). Record the results in Annexures III & IV.
• Frequency: Once in a month ± 5 working days for locations mentioned in Annexures III & IV.
• Limits:

1. The limit for air sampling of Grade A locations is < 1 cfu/m3.
2. The limit for air sampling of Grade C locations is 100 cfu/m3.
3. The Standard Limit for air sampling of Grade D locations is 200 cfu/m3.
4. The alert limit for air sampling of Grade D locations is 100 cfu/m3.
5. The action limit for air sampling of Grade D locations is 150 cfu/m3.

• Acceptance Criteria:

1. Bacterial colonies shall be within the limit.
2. Fungal colonies shall be absent.

Action plan for count exceeding alert limit:

• During routine monitoring, if the environmental monitoring count exceeds the alert limit, it has to be brought to the notice of the section in-charge, QA department, production department, and maintenance department and necessary actions have to be taken to reduce the microbial load.
• The cleaning frequency interval shall be decreased and after every cleaning activity environmental monitoring to check the bio-load.
• Continue the above procedure till the bio-load level comes below the alert limit.
• During this period production activity will be continued.

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Action plan for count exceeding action limit:

• During routine monitoring, if the environmental monitoring count exceeds the action limit, it has to be brought to the notice of the section in charge, the QA department, the production department, and the maintenance department, and necessary actions have to be taken to reduce the microbial load.
• The source of the contamination shall be investigated through deviation control. Sources could include heating, ventilating, and air conditioning (HVAC) systems, damaged HEPA filters, and changes in personnel garbing habits or working practices.
• During this period production activity will be stopped.
• Thoroughly sanitize the areas by means of Fumigation.
• Keep the areas clean and dry and sanitize the equipment with 70% IPA solution.
• Minimize the movements in the production area.
• After the above necessary actions perform environmental monitoring to check the Bio-load levels.
• Repeat the above procedure till the bio-load level comes below the alert limit.

Action plan for count exceeding standard limit:

• During routine monitoring if the environmental monitoring count exceeds the standard limit, it has to be brought to the notice of the section in charge, QA department, production department, and maintenance department, and necessary actions have to be taken to reduce the microbial load.
• Inform the manufacturing department to stop the activity in that particular area.
• Carry out the fumigation activity and proper cleaning procedure.
• Clean the area, walls, and floor, with disinfectants and frequently change the disinfectant solution.
• After fumigation and cleaning activity perform environmental monitoring to check the bio-load level.
• Continue the above procedure till the bio-load level comes below the alert limit.

ABBREVIATIONS

QC: Quality Control

QA: Quality Assurance

SOP: Standard Operating Procedure

CFU: Colony Forming Unit

SCDA: Soyabean Casein Digest Agar

SDA: Sabouraud Dextrose Agar

TAMC: Total Aerobic Microbial Count

LAF: Laminar Air Flow

TYMC: Total Yeasts & Mold Count

NMT: Not more than

RLAF: Reverse Laminar Air Flow

REVISION HISTORY

Nil

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