: QUANTITATION OF VIABLE AIR BORN

1. PURPOSE:

1.1 It is established to access quantitatively microbiological status (microbial contamination levels) of the critical environments.

2. SCOPE:

2.1 It is applicable in the pharmaceutical controlled/clean rooms.

3. RESPONSIBILITY:
3.1 Quality Control Manager
3.2 Microbiologist
3.3 Quality assurance officer
3.4 Micro-Lab.Assistant
4. PROCEDURE:
4.1 Suitable Apparatus:
4.1.1 75-mm Diameter Petri Plates
4.1.2 Aluminium Foil
4.1.3 Conical Flask 250 ml
4.2 Suitable Equipments:
4.2.1 Air Sampler
4.2.2 Incubator @ 37°C
4.2.3 Hot Air Oven
4.2.4 Autoclave
4.2.5 Colony counter
4.3 Media Required:
4.3.1 Nutrient Agar
.
4.4 Autoclavation of Apparatus:
4.4.1 Wash the petri dishes with a suitable detergent.
4.4.2 Place & dry them in a suitable Hot Air Oven, Thermostatically controlled, capable of maintaining the temperature at 80•C.
4.4.3 Sterilize the apparatus at 121 °C for about 20 minutes

4.5 Preparation / Autoclavation of Media (Nutrient Agar):
4.5.1 Dissolve required quantity of Nutrient Agar Media as directed on its label in 100 ml of distilled water by boiling.
4.5.2 Autoclave in a steam sterilizer at 15 lb pressure (121 °C) for 18 to 20 minutes
4.5.3 Congeal a uniform (25ml) layer of the media in in Petri dishes under Laminar Air Flow Hood & let it to solidify, cover and invert the plates.
4.5.4 Incubate for at least 24 hours to check for any growth.

4.6 Methodology:
4.7.1 Define the Sampling sites in the critical area.
4.7.2 Charge the battery of the air sampler.
4.7.3 Adjust the sterilized Nutrient Agar Plate in the Sampling Equipment under LAF Hood and transfer the charged equipment in the sampling sterile area.( Keep one Petri plate unexposed as negative control.)
4.7.4 Switch on the Sampler to suck the air through perforated lid and to fall it on the Agar Plate in aregular pattern.
4.7.5 Transfer the Device to the Microbiology Laboratory aseptically.
4.7.6 Discharge the Plate & incubate @ 35°C for 48 hours.
4.7.7 After completion of incubation time calculate the colony forming units (CFUs) for their acceptance criteria.(The number of sampling Plates may Be increased according to the running situation)