Guideline and Formats for Microbiological Environmental Monitoring (EM) of various area like in Production, Microbiology laboratory, Dispensing & Sampling Area, and Manufacturing Area etc.
Microbiological Environmental Monitoring (EM)
1.0 Objective :
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- To lay down a procedure for microbiological of environmental monitoring.
2.0 Scope :
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- This SOP is applicable for microbiological environmental monitoring of area in Production, Microbiology laboratory, Dispensing & Sampling Area, and Manufacturing Area (Tablet).
3.0 Procedure for Environmental Monitoring (EM) :
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Introduction – Environmental Monitoring:
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- The routine microbiological environmental monitoring of environment in classified environmental zones contributes to the assurance of product quality.
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- Any classified area where manufacturing, processing, packing, holding or testing of products take place needs to be maintained in a clean and sanitary conditions.
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- This is achieved by restricting the entry of particulate contamination in to these areas.
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- The environment of such classified environmental zones shall be monitored for the level of microbial contamination by the controls described hereunder.
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- The types of controls and the relevant acceptance criteria shall depend upon the grades of classified environmental zones.
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Method for Environmental Monitoring (EM):
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- There are two methods for microbiological environmental monitoring of area
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- Air Sampling Method (Active sampling)
- Air Sampling Method (Active sampling)
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- Settle Plate Method (Passive sampling)
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Air Sampling Method (Active Sampling) :
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- Preparation of Petri dishes:
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- Prepare petri dishes of Soyabean Casein Digest Agar (SCDA) as per SOP for Media Preparation and perform environmental monitoring as per annexure.
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- Use pre-incubated Petri dishes for environmental monitoring.
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- Use petri dishes with absolutely no sign of microbiological contamination at the end of incubation.
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- Sampling head of air sampler shall be sterilized by autoclaving and mop the body of air sampler with 70% IPA.
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- Take the air sampler at the respective location and assemble it.
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- Write the given below details on petri dishes before assemble it in air sampler
Name of Media: ………………………………………………..
Plate preparation Date: ……………………………..………….
Area: ………………………………………..……………………
Date of Exposure: ………………………………………………
Sampled By: …………………………………………….………
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- Run the air sampler unit flows the required volume of air on the plate as per given below Table.
GMP- EU | ISO Class | Air Sampling Volume |
A | ISO 5 | 1000 Liter |
B | ISO 6 | 1000 Liter |
C | ISO 7 | 1000 Liter |
D | ISO 8 | 1000 Liter |
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- After sampling take the plates to microbiology laboratory for incubation.
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- Incubate the petri dishes at 22.5±2.50C for 72 hrs for fungal growth followed by at 32.5±2.5°C for 48 hrs for Bacterial Growth.
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- Record the result of the respective area in respective annexure
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Settle Plate Method (Passive sampling) :
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Principle:
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- Petri dishes filled with nutritive agar medium are exposed in the environment and the airborne microorganisms settle down on the surface of the agar.
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- The plates are incubated for optimum growth of microorganisms and the colony forming units (CFU) developed at the end of incubation are counted.
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- The number of CFU developed on the settle plate at the end of incubation indicates number of viable microorganisms per 90 mm diameter of the exposed Petri dish.
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Preparation of Media Plates:
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- Prepare petri dishes of Soyabean Casein Digest Agar (SCDA) as per as per SOP for Media Preparation.
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- Use pre-incubated petri dishes for environmental monitoring.
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- Use petri dishes with absolutely no sign of microbiological contamination at the end of incubation.
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Sampling & Observations – Environmental Monitoring (EM):
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- Mope the SS container with 70% IPA solution and load the petri dishes carefully.
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- Close & carry the container to the respective areas.
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- Ensure that the area under environmental monitoring is clean and sanitized as per defined procedure.
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- Ensure that the Temperature/Humidity in the area under microbiological environmental monitoring is within the specified limit.
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- Place the petri dishes containing SCDA as prepared above carefully number wise, at the marked places defined in respective Annexure.
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- Do not expose one petri dish and take it as negative control.
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- Carefully lift the lid of petri dish and place it gently on stand near the exposed plate thereby exposing the entire agar surface.
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- Leave the petri dishes in the exposed condition for 4 hours.
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- After completion of exposure gently place the lids over the exposed petri dishes and load them back in the drum, bring them out of the controlled area.
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- Incubate the petri dishes at 22.5±2.50C for 72 hrs for fungal growth followed by at 32.5±2.50C for 48 hrs for Bacterial Growth.
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- Stick working culture on one plate and take it as positive control and treat in the same manner as above.
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- Report the results in the specified format. The relevant formats are Annexure.
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Selection of location for plate exposure by settle plate method:
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- For core area where the material is exposed the number of plate shall be calculated based on √A, where A is room area in m2.
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- The plate shall be exposed on return air risers and other plates shall be exposed on corners at a angle of approx 45° in SS stand.
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- For ancillary areas the plate shall be exposed on return air risers and additional plates in corners
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- In room where diffusers are installed plates shall be exposed in corners.
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Allotment of plate ID No.
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- The plate No. shall be linked with the riser ID No. and shall have the same ID No. as of riser, on which the plate to be exposed.
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- The plates which are not exposed on risers the ID No. shall be allotted as II-NN. Where II is AHU No. and NN is serial No. in continuation of Riser ID No.
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- Limits: as defined in respective annexure (s).
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Frequency:
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Area | Frequency |
OSD | (Twice in a month), alternate Active and Passive. |
Oral Liquids | (Weekly), alternate Active and Passive. |
Dispensing & Sampling Area | (Weekly), alternate Active and Passive. |
Microbiology Laboratory | (Weekly) Active and Passive both. |
Manufacturing Area (Tablet) | (Weekly), alternate Active and Passive. |
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- Trend frequency: Trend of settle plate and air sampling frequency is every six month.
Area | Frequency |
OSD | Every Six Month |
Oral Liquids | Every Six Month |
Dispensing & Sampling Area | Every Six Month |
Microbiology Laboratory | Every Six Month |
Manufacturing Area (Tablet) | Every Six Month |
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- Any OOS results shall be handled following SOP – Handling of OOS Results.
4.0 Reference (S)
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- Schedule “M” of Drugs and Cosmetic act section 4 of part 1 A.
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- ISO 14644 – I Clean room and associated controlled environments.
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- Chapter 6 : Quality control of EU guidelines for Medicinal products for Human and Veterinary use Volume 4,
5.0 Annexure – Environmental Monitoring :
Annexure- Environmental Monitoring of using settle plates
Method : Settle Plate
Date of Sampling: ……………. Plate Exposure Time:……………
Plate Collection Time : ……….. Exposed By: …………………….
Media Used: Soyabean Casein Digest Agar Date of Completion: ……………
Incubation Temperature: 22.5±2.50C for 72 hrs and 32.5±2.50C for 48 hrs.
Exposure Time: ________ Frequency: _______
Location |
Plate ID No. |
Total Microbial Count ( cfu/ 90 mm plate / 4 hrs) |
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Observation |
Limit | |||||||
Alert | Action |
Max. |
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