Environmental monitoring of LFH

1. Purpose:-

To describe the method of environmental monitoring of Laminar Air Flow Hood (LFH).

2. Scope:-

It is 15.04.2015 applicable for all Laminar Air Flow Hoods (LFHs) present in all clean rooms of Production and Microbiological Lab.

3. HSE Statement:-

Proper gowning technique should be followed.

4. Responsibilities:-

i) Manager Quality Control is responsible to ensure that procedure & formats are followed entirely as approved.

ii) Microbiologist is responsible to perform the test.

5. Materials:-

5.1 Anemometer

5.2 Non-viable Laser Air Particle Couter.

5.3 70% IPA filtered

5.4 Fine dusters

5.5 SS-tray

5.6 Sterile Swabs and Sterile phosphate buffer pH 7.2

5.7 Surface Air sampler

5.8 Sterile and pre-incubated Tryptic Soya Agar plates

6. Definitions:-

6.1 LFH: - Laminar airflow cabinets are designed to direct particles away from the work surface

(This is often supported by airflow or smoke studies).The term 'LAF' is becoming increasingly

replaced by UDAF or uni-directionalair-flow

6.2 Environmental monitoring:

A documented series of sampling and testing in controlled environments in order to

Demonstrate conformance to a series of pre-set limits or for trends.It is distinct from environmental control.

6.3 Air Sampling:-

The collection a sample of air; can be physical (the collection and estimation of the number of airborne particles, normally called particle counting) or viable (the estimation of the number of viable micro-organisms are either deposited onto a settle plate - passive sampling - or collected from a defined volume of air - active / volumetric air-sampling using an air-sampler.

6.4 Air velocity: -

A value indicating the speed of the air movement in a clean room or clean zone. It is an indicator of the ability of a unidirectional-flow clean room to purge itself.

6.5 Particle counter:-

A device which measures the number of airborne particles of a given size by passing

a sample of air through a focused light source. The presence of a particle causes dispersion of the

light and the presence of a particular size can be detected using pre-defined criteria. At BPL two

sizes of particles are measures: 0.5μm (a close approximation of the size of a bacterium)

and 5.0μm (a close approximation of the size of a human skin cell).

7.Flow Chart:-

8. Description:-

8.1 Procedure:-

8.1.1 Microbiological Monitoring of LFHs: -

8.1.1.1 Perform surface swab test of following parts of LFH in following order as per SOP

8.1.1.2 Sheets, covers or walls

8.1.1.3 Side Grills of HEPA filter of LFH

8.1.1.4 Working bench or table

8.1.1.5 Machine parts or other accessories

8.1.1.6 Expose settle plates for 4 hours under LFH at designated points as per SOP.

8.1.1.7 Perform surface air sampling of LFH as per SOP.

8.1.2 Determination of Airflow velocity:-

8.1.2.1 Check the battery of anemometer. If low, replace the battery. Also check the anemometer for its

proper working.

8.1.2.2 Wipe the anemometer with duster dipped in 70% filtered IPA. Care should be taken with probe whilst wiping the probe.

8.1.2.3 Place the anemometer in SS-tray already wiped with 70% filtered IPA.

8.1.2.4 Transfer the SS-tray to the area to be tested and place the SS-tray in pass through under UV- light for at least 20-30 minutes.

8.1.2.5 Take the anemometer out of SS-tray and again wipe with duster dipped in 70% filtered IPA.

8.1.2.6 Divide the work zone of LFH into 5 equal parts as shown below:

1		2
	3
4		5

8.1.2.7 Orient the probe perpendicular to the air flow at a distance of not more than 150mm (6 inches) from the HEPA filter or 1 foot above the work surface. All test positions should be within unobstructed airflow. Measure the airflow velocity at each test position.

8.1.2.8 For unidirectional airflow system, take the readings form single HEPA filter at designated points shown above. For non-unidirectional airflow system, take the readings form each HEPA filter separately at designated points shown above.

8.1.2.9 Orient the probe at the specified location until a stable reading is obtained and record the average reading.

8.1.3 Non-viable Air Particle Count:-

8.1.3.1 Charge and check the non-viable particle counter for its proper working.

8.1.3.2 Wipe the non-viable particle counter and its probe with duster dipped in 70% filtered IPA.

8.1.3.3 Place the equipment in SS-tray already wiped with 70% filtered IPA.

8.1.3.4 Transfer the SS-tray to the area to be tested and place the SS-tray in pass through under UV- light for at least 20-30 minutes.

8.1.3.5 Take the equipment out of SS-tray and again wipe with duster dipped in 70% filtered IPA.

8.1.3.6 Set up the non-viable particle counter by attaching the probe to non-viable particle counter in its port.

8.1.3.7 Record the name of operator(s) working in the clean air device being tested and their activity at the time of testing.

8.1.3.8 Position the probe isokinetically in relation to the air flow (facing the air flow). Particle counter should be positioned at 1 foot above the working level.

8.1.3.9 Run the particle counter. Observe the first count to ensure the particle counter is operating correctly and discard the reading.

8.1.3.10 Take the consecutive three reading for 0.5μm particles by moving the particle counter in entire area so that all area is sampled for non-viable particle count. Sampling time must be 1 minute for each reading If there are more than one HEPA filters in a LFH, take the separate readings for each filter as described above

8.1.3.11 Take the average of readings for both particles.

8.1.4 Air Sampling:-

8.1.4.1 Charge and check the air sampler for its proper working prior to use..

8.1.4.2 Wipe the air sampler with duster dipped in 70% filtered IPA.

8.1.4.3 Place the equipment in SS-tray already wiped with 70% filtered IPA.

8.1.4.4 Transfer the SS-tray to the area to be tested and place the SS-tray in pass through under UV- light for at least 20-30 minutes.

8.1.4.5 Take the equipment out of SS-tray and again wipe with duster dipped in 70% filtered IPA.

8.1.4.6 Open the lid of air sampler and adjust a prepared sterilized Agar plate within the grooves of air sampler and screw up the lid.

8.1.4.7 Turn on the air sampler it automatically turns to operating page within 3 seconds.

8.1.4.8 Now input the sampling air volume which is 1cubic meter (Equivalent to 1000liter)

8.1.4.9 Push ENTER, it starts to sample.

8.1.4.10 Take the samples of air from three different locations by moving the air sampler in entire area under LFH so that all area is sampled for viable particle count. If there are more than one HEPA filters in a LFH, take the separate readings for each filter as described above

8.1.4.11 Incubate these plates at 32.5⁰C for 48-72 hrs and note down the results.

8.2 Specifications:-

8.2.1 Microbiological Monitoring: Area under LFH should comply with specifications of surface swab test; settle plate method and surface air sampling as per SOP.

8.2.2 Velocity: The air flow velocity of LFH should not be less than 90fpm (0.45m/s) + 20%.

8.2.3 Non-viable Air Particle Count: NMT 3500 particles of 0.5-5.0μm should be observed under LFH.

8.2.4 Viable Particle count: Less than 1cfu.

8.3 Frequencies:-

8.3.1 Settle Plate Method: Each Operating Shift or as and when required

8.3.2 Surface Swab Test: Monthly or as and when required

8.3.3 Surface Air Sampling: Monthly or as and when required

8.3.4 Airflow Velocity: Monthly or as and when required

8.3.5 Non-viable Air Particle Count: Monthly or as and when required

9. Records:-

9.1 Aseptic Area Monitoring (SPM and SAS) – Ampoule and IV Infusion Area

9.2 Aseptic Area Monitoring (SPM and SAS) – Vial Filling Area

9.3 Area Monitoring of Sterility Testing Suit

9.4 AC/CFM/LV Monitoring Log

9.5 Non-viable Air Particle Count Report

10 References

10.1 ISO 14644-3:2005, Clean rooms and associated controlled environments.

10.2 FDA, Guidance for Industry, Sterile Drug Products Produced by Aseptic Processing — Current

Good Manufacturing Practice, September 2004.

10.3 Syed Imtiaz Haider, Validation Standard Operating Procedures A Step by Step Guide for Achieving Compliance in the Pharmaceutical, Medical Device, and Biotechnology Industries, 2^nd edition, Taylor & Francis Group, LLC, USA. 2006

10.4 Guidelines on the Test Methods for Environmental Monitoring for Aseptic Dispensing Facilities, Produced by: A Working Group of Scottish Quality Assurance Specialist Interest Group, Feb 2004.