Environmental Monitoring Procedure and its Sop

Environmental Monitoring; Purpose

Environmental Monitoring; To describe the procedure for Environmental monitoring.

Scope

Environmental monitoring in  XX Pharmaceuticals Ltd

Definitions

Environmental monitoring:

Monitoring of viable and non-viable quality of a controlled environment

Particulate count:

[] Enumeration of non-viable particulate of specific size from a particular volume of air of a controlled
Environment.
[] Settle Plate: Exposure of petri-plates of nutrient media in a controlled environment to estimate viable
Microorganisms from the environment.
[] TSA: Tryptone Soya Agar
[] CFU: Colony Forming Unit

Responsibilities

The roles and responsibility is as follows:

Lab. Attendant

Room preparation for Test

Executive, Microbiology

Carry out the test and incubation & documentation

Sr. Executive, Microbiology

To ensure test, incubation, report checking, document preservation & application of sound technical information.

Head of Plant

Review of the SOP & the relevant technical information is applied.

Head of Quality Assurance

Take initiative to Approve of this SOP.

Procedure:

Instructions

• Don’t rub during contact plate sampling.
• Use the sterile filter holder.
• Place the filter paper on the filter holder carefully under laminar airflow.
• Try to avoid unwanted personal contamination during air sampling.
• Monitoring to be carried out before production hours. (Sterile products)

Non-viable Particle monitoring:

• Bring the laser particle counter into the specific monitoring area.
• Enter into clean room, wearing approved designated dress. When entering into the clean area, before taking reading make sure that all the doors remain closed.
• Operate the Particle counter following approved SOP.
• Take count of different several position for each room as indicated in sampling point. Count the number particles (5µ and 0.5 µ) form each sampling point. The average of the counted particle indicates the total particles of a room.

The minimum sampling time should be as per following formula (Following EN ISO 14644-1)

Vs=(20/Cn,m ) x 100

Where,

• Vs = is the minimum single sampling volume per sampling point, expressed in liter.
• Cn,m = is the class limit (number of particles per cubic meter) for the largest considered
  particle size, specified for the relevant class.
• 20 = is the defined number of particles that could be counted if the particle concentration
  were at the class limit.

Specification: Follow Annexure-III.

Write down the result in the format of Annexure-I.

 

Viable count:

Settle Plate

• Prepare, sterilize & dispense the media TSA into the petriplates and pre-incubate the plates.
• Check the pre-incubated plates for any evidence of microbiological contamination under the LAF bench.
• Discard the plates contains microbiological growth.
• Decontaminate the external surface of petriplates with sterile cloth /cotton soaked in a sanitizing agent (70% IPA).
• Place required number of petriplate in a sterilized /sanitized SS container; close the lid of SS container.
• Transfer SS box to the area to be monitored.
• Enter respective area as per the SOP of Entry and Exit procedure.

Mark Petri plates with the following details-

1. Name of the sampling point
2. Room No.
3. Date of exposure

• Place petriplates on corresponding designated plate exposure area and remove
  the upper lid of the petriplates. Note the beginning time of the exposure and write it on
  the plate.
• Place upper lid on edge of petriplates in slanting position.
• Expose media plates at sampling point for Maximum 4 hours.
• After completion of exposure, close petriplates with lid.
• Collect petriplates in the SS container and bring back exposed plates to microbiology Laboratory.
• Incubate all exposed petri plates in inverted position in laboratory Incubator.
• Incubate at (22.5 ± 2.5)°C for mold and Yeast for 72 hours followed by another 48 hours for bacteria at (32.5 ± 2.5)°C. After incubation count the number of colony forming units (CFU) per plate per sampling point with colony counter.

• Incubate an unexposed media filled petriplates along with the exposed plates and mark it as negative control.
• Count the number of Colony Forming Units (CFU) per plate per location with colony counter. The average of colony indicates total CFU of a room.

Write down observation in the format attached as Annexure-II.

Air Sampling:

• Set instrument for desired sampling time & flow rate as per Approved SOP.
• Take air sampler to location where air is to be sampled, hold it with filter facing at required direction and take air sample following approved SOP.
• Collect filter under laminar airflow & place it inside on petriplates containing TSA[Tryptone Soya Agar ] media.
• Incubate all exposed petri plates in inverted position in the microbiology laboratory Incubator. Incubate at the (22.5 ± 2.5)°C for mold & Yeast for 72 hours followed by another 48 hours for bacteria at (32.5 ± 2.5)°C. After incubation count the number of colony forming units (CFU) per plate per location with colony counter.
• Follow sampling location.
• Record observation in format attached as Annexure-II.
• Calculate number of organisms per cubic meter of air. Average colony indicates total cfu of a room.

Surface monitoring (By swab sampling):

• Use sterilized swab or sterilize the swab in the autoclave.
• Place required number of sterilized swab sticks with tube containing 5 ml of sterile saline
  solution in a Sterilized SS container & tightly secure lid of the SS container.
• Transfer SS box to area to be monitored.
• Enter respective area as per SOP of entry and exit procedure.

Remove swab sticks with tubes from the SS box & take it to the location to be monitored and Mark tube with following details:

1. Location number or Name of the location
2. Swab No.
3. Date of sampling

• Hold swab stick from bottom & place tip on surface to be monitored. Gently wipe the swab bi-directionally to cover an area about 25 cm2.
• Perform swab sampling for each defined locations.
• Keep swab sample standing in the SS container & bring it to Laboratory.
• Gently vortexes swab sampling tube containing swab stick & pour contents in filter holder funnel and filter it. Uses the respective SOP.
• Rinse tube with 3 x 10 ml of sterile saline solution.
• Filter test sample under partial vacuum.
• Rinse Funnel with the portions of sterile Purified Water. This flushes residue from walls of funnel & helps to secure a uniform distribution of colonies on filter surface. Upon completion of rinse & filtration process, shut off vacuum.
• Transfer membrane filter with sterile smooth-tip forceps on to Microbial content test agar (TSA) plate.
• Place filter with a rolling motion to avoid entrapment of air.
• Keep a negative control by filtering 100 ml Purified Water(Sterile) before filtering actual test sample.
• Pass 20 to 30 ml of sterile Purified Water through funnel, between different test samples when the same funnel used for multiple sample.
• Incubate all exposed petriplates in inverted position in microbiology laboratory Incubator. Incubate at (22.5 ± 2.5)°C for mold and Yeast for 72 hours followed by another 48 hours for bacteria at (32.5 ± 2.5°)C. After incubation count number of colony forming units (CFU) per plate per location with colony counter.
• Count number of colony forming units (CFU) per plate per location with colony counter.

Note down the observation in the format attached as Annexure-II

Surface monitoring (By contact plate):

• Fill Petridishes with TSA culture medium & pre-incubate.
• Take off the lid of plates.
• Invert & press agar surface for 10 seconds onto surface to be examined.
• Replace lid & mark plate with appropriate data.
• Clean sampling area on surface in order to remove any remaining of agar.
• Return plates to laboratory.
• Incubate all exposed petriplates in inverted position in microbiology laboratory Incubator. Incubate at (22.5 ± 2.5)°C for mold and Yeast for 72 hours followed by another 48 hours for bacteria at (32.5 ± 2.5°)C. After incubation count number of colony forming units (CFU) per plate per location with colony counter.
• Take count of visible colonies within mean of plate
• Express results as CFU per contact plate.
• Perform appropriate identification of germs found by surface monitoring..Classified area should free of pathogenic microorganism.

Note down the observation in the format attached as Annexure-II.

Sampling Location:

Determine Number of sampling location by following formula.

N_L    = √A

Where,

N_L   = the minimum number of sampling locations, rounded up to a hole number.

A= Area of the clean room or clean air controlled space in m^2.

^{Environmental Monitoring Procedure as per Pharmacopeia}

Download All Annexure Here: Environmental Monitoring

• Annexure-I: Environmental Monitoring (Non-Viable Particle Count) Test Report
• Annexure-II: Environmental Monitoring (Viable Particle Count) Test Report
• Annexure-III: Environmental Monitoring specification
• Annexure-IV: Log book of environmental monitoring

Schedule: Environmental Monitoring

• Environmental monitoring Test Schedule