ENVIRONMENT MONITORING

Procedures:
Equipments Required:
Air Sampler
Incubators (20ºC to 25ºC and 30ºC to 35ºC)
LAF/BSC Unit
Autoclave
SS stencil
SS containers to carry plates
Materials Required:
Filtered 70% IPA
Sampling kit
Sterilized Air Sampler Heads
Sterilized wipes
Media required (media prepared from dehydrated media or ready to use media
plates):
a) Pre incubated Air Sampler Plates (SCDA with 1% glycerol)
b) Pre incubated Settle Plates (SCDA with 1% glycerol)
c) Contact plates ( SCDA with Polysorbate 80 & Lecithin)
d) Sterile swab and sterilized phosphate buffer

Preparation for Sampling
Prepare & sterilize:
1. SCDA + 1% glycerol (passive and active air sampling)
2. SCDA + neutralizer (Surface monitoring)
3. SCDA + 1% glycerol + neutralizer (Finger dab)
Pre incubate for 48 ± 2 hours at 30°C to 35°C
(If used beyond 2 days, stored at 2°C-8°C then pre incubate)

Examine plate (dryness/cracks/contamination)

wipe by using 70% IPA. Check cleanliness of SS stand. come out by following clean room exit procedure Passive Air Sampling (Settle Plates) Place plate in sampling location. sampling point and personnel name) Sanitize inner & outer sampling kit with sporocidal and followed by 70% IPA Keep media in sampling kit. Before carry to pass box. location. make sure each point in checklist sampling is completed. Enter clean room with clean room gowning procedure After completing.Label & pack for sampling point and one negative control (Date & time exposure. Expose plates by placing on the upper part of the SS stand and the .

Place plate on air sampler & close with perforated head air sampler. Avoid to pass the hand over perforated head. Check result. record and verify. close plate and incubate all the plate with negative control plate at 20°C to 25°C for 3 days. remove the perforated head by holding the edges.lid of the plate is placed facing downwards on the lower part of SS stand. Put lid on screen of air sampler. record and verify. . Check result. Take the plate and incubate all the plate with negative control plate at 20°C to 25°C for 3 days. After 4 hours. Expose plate at sampling point area at least 4 hours. After sampling over. Active Air Sampling Check battery power for air sampler to ensure battery status before use along with calibration status. Sanitize and sterilize all sampling equipments. then shift 30°C to 35°C for 2 days. Draw 1000 liters of air onto air sampler in 10 minutes with 1 minute of delay time. then shift 30°C to 35°C for 2 days.

Spread evenly without twisting/sliding Then. close the plate with lid. record and verify. Clean the surface by wipe with 70% IPA Incubate all the plate with negative control plate at 20°C to 25°C for 3 days.Surface Sampling (Contact Plate) Before sampling. Check result. then shift 30°C to 35°C for 2 days. make sure surface is dry Remove lid & place agar surface for 5-10 seconds. Surface Sampling (Swab Technique) Open the sterile swab aseptically and dip in 10m sterilized phosphate buffer saline Apply slight pressure and hold the swab approximately at 30° angle Swab the area in close parallel strokes covering about 5cm X 5cm area followed by swabbing again at right angles to the previous swabbed area Perform this activity by using a sterilized SS stencil of 25cm2 where applicable .

discard the swab Apply the vacuum and filter the contents Add about 100ml of the sterilize 0. dip the swab in the sterile phosphate buffer and close the lid of tube Label the swab sample with location name. start exercise again with new filtration unit and membrane filter.1% peptone water. fit it into the manifold apparatus and place the sterilized filter membrane by using sterilized forceps by opening the upper portion of the filtration unit and placing the membrane on the inner portion of the filtration unit Close the upper portion of filtration unit and add about 100ml of the sterilized 0. repeating the exercise twice . Gently swirl entire quantity (10ml) of phosphate buffer and transfer the phosphate buffer into filtration unit.Then.1% peptone water into filtration unit and filter it. Apply vacuum to filter the contents If any leak happen. sample details and date of sampling on the tube Unload the tubes containing swab and phosphate buffer under LAF in QC Lab Take the sterilized filtration unit.

then shift 30°C to 35°C for 2 days. Check result. record and verify. Check Result Personal Monitoring (Finger dabs) Taken on 90mm petri plates containing neutralizer by having gentle impression of all the four gloved finger tips of one gloved hand. Incubate all the plate with negative control plate at 20°C to 25°C for 3 days. Disinfect the hands immediately with 70% IPA and discard the glove. date of analysis and date of release Incubate all the plate with negative control plate at 20°C-25°C for 3 days. then shift 30°C-35°C for 2 days. fist and thumb impression onto one plate followed by other gloved hand on to other plate For filling and fitration perform the finger dabbing and gown monitoring near the filling personnel exit air lock.Open the upper portion of the filtration unit and pick the membrane filter with sterilized forcep and transfer to the SCDA media with neutralizer which labeled with swab details. For formulation personnel perform the dab monitoring near the personnel exit airlock. .

right and left of chest. point of sampling and date of sampling.Personal Monitoring (Gown) Press agar surface gently against the gown of the forehead. . then shift 30°C to 35°C for 2 days. This is done per batch and at the end of the activity/shift for each operator. Check result. record and verify. Hold for 10 to15 seconds. Disinfect the gown by wiping the sampled area with a lint free cloth wetted with 70% IPA. Swab also can be used and the same procedure is applicable as mentioned in surface monitoring for swabbing and analysis. load the samples into SS container and carry as per exit procedure. Incubate at 20°C to 25°C for 3 days. right and left of forearm and right and left of leg using separate contact plates for each sampling point on the gown. After completion. perform this activity of 25 cm2 per sampling point. Label the contact plate with operator name.

Incubation and Observations: a) Incubate all the plates along with the control plates in an incubator at 20°C to 25°C for 3 days. then shift 30°C to 35°C for 2 days. Note : The negative control plate should not show any microbial growth at the end of the incubation period. b) Observed the incubated plates during 3 days (while shifting) and after 2 days for microbial counts. A failed negative control will lead to an investigation. . record the results in respective formats.