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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 :

• • To lay down a procedure for microbiological of environmental monitoring.

2.0   Scope :

• • 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) :

• • Introduction – Environmental Monitoring:

• • The routine microbiological environmental monitoring of environment in classified environmental zones contributes to the assurance of product quality.

• • Any classified area where manufacturing, processing, packing, holding or testing of products take place needs to be maintained in a clean and sanitary conditions.

• • This is achieved by restricting the entry of particulate contamination in to these areas.

• • The environment of such classified environmental zones shall be monitored for the level of microbial contamination by the controls described hereunder.

• • The types of controls and the relevant acceptance criteria shall depend upon the grades of classified environmental zones.

• • Method for Environmental Monitoring (EM):

• • There are two methods for microbiological environmental monitoring of area

• • • Air Sampling Method (Active sampling)

• • • Settle Plate Method (Passive sampling)

• Air Sampling Method (Active Sampling) :

• • Preparation of Petri dishes:

• • Prepare petri dishes of Soyabean Casein Digest Agar (SCDA) as per SOP for Media Preparation and perform environmental monitoring as per annexure.

• • Use pre-incubated Petri dishes for environmental monitoring.

• • Use petri dishes with absolutely no sign of microbiological contamination at the end of incubation.

• • Sampling head of air sampler shall be sterilized by autoclaving and mop the body of air sampler with 70% IPA.

• • Take the air sampler at the respective location and assemble it.

• • 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: …………………………………………….………

• • Run the air sampler unit flows the required volume of air on the plate as per given below Table.

• • After sampling take the plates to microbiology laboratory for incubation.

• • 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.

• • Record the result of the respective area in respective annexure 

• Settle Plate Method (Passive sampling) :

• • Principle:

• • Petri dishes filled with nutritive agar medium are exposed in the environment and the airborne microorganisms settle down on the surface of the agar.

• • The plates are incubated for optimum growth of microorganisms and the colony forming units (CFU) developed at the end of incubation are counted.

• • 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.

• • Preparation of Media Plates:

• • Prepare petri dishes of Soyabean Casein Digest Agar (SCDA) as per as per SOP for Media Preparation.

• • Use pre-incubated petri dishes for environmental monitoring.

• • Use petri dishes with absolutely no sign of microbiological contamination at the end of incubation.

• • Sampling & Observations – Environmental Monitoring (EM):

• • Mope the SS container with 70% IPA solution and load the petri dishes carefully.

• • Close & carry the container to the respective areas.

• • Ensure that the area under environmental monitoring is clean and sanitized as per defined procedure.

• • Ensure that the Temperature/Humidity in the area under microbiological environmental monitoring is within the specified limit.

• • Place the petri dishes containing SCDA as prepared above carefully number wise, at the marked places defined in respective Annexure.

• • Do not expose one petri dish and take it as negative control.

• • Carefully lift the lid of petri dish and place it gently on stand near the exposed plate thereby exposing the entire agar surface.

• • Leave the petri dishes in the exposed condition for 4 hours.

• • 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.

• • 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.

• • Stick working culture on one plate and take it as positive control and treat in the same manner as above.

• • Report the results in the specified format. The relevant formats are Annexure.

• • Selection of location for plate exposure by settle plate method:

• • 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.

• • 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.

• • For ancillary areas the plate shall be exposed on return air risers and additional plates in corners

• • In room where diffusers are installed plates shall be exposed in corners.

• • Allotment of plate ID No.

• • 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.

• • 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.

• • Limits: as defined in respective annexure (s).

• • Frequency:

• • Trend frequency: Trend of settle plate and air sampling frequency is every six month.

• • Any OOS results shall be handled following SOP – Handling of OOS Results.

4.0   Reference (S)

• • Schedule “M” of Drugs and Cosmetic act section 4 of part 1 A.

• • ISO 14644 – I Clean room and associated controlled environments.

• • 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: _______