HVAC TEST PROCEDURES

o Ensure that the HVAC is operational prior to start of any testing or observation.

Air Velocity and Air Changes

Objective
o To ensure that the room is supplied with adequate amount of air, and wherever
specified, the number of air changes is as per the specification
Scope
o This test is applicable to those HVAC installations where the air velocity and air
changes is specified and is critical for maintaining the cleanliness class
Test Equipment
o Calibrated thermal anemometer or vane type anemometer shall be used
Procedure
o Measurement of air velocity
• Ensure that the all doors of the room are properly closed.
• Remove the supply air diffuser (if any) before starting the test.
• Velocity at the inlet air grills shall be measured at 5 points as
illustrated in Fig -1 in a plane parallel to filter face plane and at a
distance of about 6 inches (~ 150 mm) from the filter/opening
face.
o Fig. 1

•
The velocity shall be measured for at least 10 seconds from
each point.
o Measurement of number of air changes/hour

• Volume of air supplied through the duct is calculated by

multiplying the average velocity with the surface area of opening.
The total volume of air supplied into the area is derived by
summing up the volume of air supplied through all inlet ducts of
the area.

o Formula for calculation

o Step I: Calculate the air velocity at SA

o For single grille – Mean air velocity (m/s) at single filter/grille ------------ (A).

o Step II: Calculate the air volume at SA

 o Air supply volume (m3/s) for HEPA/grille = (A) m/s x HEPA/grille area m2 --------
(B1).

o Step III: Calculate the room supply volume

o Total volume supplied in the area (B) = B1 + B2 + B3 ------------------- B n in the

area.
o Step IV: Calculate the air changes for the room
o Air changes per hour is calculated by dividing the total air quantity supplied into
o Area by the volume of the room, by using the following formula

 D = ∑B ×3600
R
 D = no. of air changes
 B = Air supply volume (m3 / s)
 R = volume of the room (m3)
• = factor (for air change per hour)
 Differential Pressure test
Objective
o The objective of this test is to verify the capability of the complete installation to
maintain the specified pressure difference between the installation and
surroundings and between separate spaces within the installation
Scope
o This test is applicable to those HVAC installations where the differential pressure
across the surrounding environment is specified and is critical for maintaining the
cleanliness class
Principle
o Differential pressure gauge shall be used to measure the air pressure difference
between two areas
Test Equipments
o Calibrated inclined manometer or digital manometer or mechanical manometer

Procedure
• Ensure that all the doors of the area are closed.
• Ensure that all related HVACs are operating.
• Ensure that all the manometers / Magnehelic gauge are
calibrated.
• Ensure that after the balancing, position of the dampers are fixed
and marked.
• Measure and record the pressure difference between the
enclosure cleanroom and any surrounding ancillary environment
or between any surrounding ancillary environment and the
external environment. If no surrounding ancillary environment is
present, the pressure difference between the enclosure
cleanroom and external environment should be measured and
recorded.
• Take the manometer reading of all the rooms avoiding any
parallax error for 3 consecutive days (shift) at 1-hour intervals.
HEPA Filter leakage test
Objective
o To ensure that the HEPA filter installation has been done properly and the
installation qualifies the filter integrity test as per the pre-designed specification.
Scope
o This test is applicable to HVAC with terminal HEPA filters
Principle
o A generated polydisperse aerosol is added to the natural upstream to achieve
the required challenge concentration. The aerosol material may include but not
limited to Dioctyl 4hthalate (DOP), or Poly alpha olefin (PAO)
Test Equipment

• Aerosol Photometer (Detector/Scanner) – Light scattering type

with threshold sensitivity of at least 10-3mg/liter. Capable of
measuring concentration in range of 20 – 100 mg/m3 and with air
sample flow rate of 1ft3 ±10% per min.
• Aerosol Generator equipped with Laskin type nozzles

Procedure
• Ensure that the air velocity is established before the filter
leakage test is started.
• Position the aerosol generator to introduce an aerosol challenge
upstream of the HEPA filter to a concentration of 20-100 mg/m³
of air by opening appropriate number of nozzles.
• Measure upstream concentration of aerosol by using upstream
port.
• Adjust the photometer’s Gain / Span control for a full-scale
deflection on 100% range.
• Scan the downstream side of the HEPA filter, its perimeter, the
seal between the filter frame and grid structure including its joints
using overlapping strokes with the photometer probe.
• The photometer probe should be about 1 inch from the surface
at a transverse rate not more than 10ft/minute with a sample flow
rate of 1cft/min ± 10%.
• If any leak is more than the specified, the above test should be
repeated after taking the recommended corrective action.
• Attach the certificates/ reports of aerosol testing to qualification
activity.
 Air Flow Visualization (Non-unidirectional flow)
Objective
o To objective of the airflow visualization is to confirm spatial and temporal
characteristics of airflow in the installation and that the maintenance of
cleanliness of the area is effective
Scope
o This test is applicable to those HVAC installations where the airflow direction is
non-unidirectional
Principle
o The flow visualization shall be confirmed either by
• Tracer thread method
• Tracer particle generator (Fogger / Titanium tetrachloride fumes shall be used for
depicting the air flow pattern)
o The profiles are recorded by storage devices such as still photography, video
cameras, tapes or disks
Test Equipments
• Tracer thread
• Fogger or Titanium tetrachloride or equivalent
• Imaging equipment (Video recording or camera)
Acceptance criteria
• The airflow direction should be logical from fresh air inlet to
return air duct.
• The airflow pattern at door should be from positively pressurized
zone to negatively pressurized zone.
Procedure (Tracer thread)
• Ensure that the pressure diffentials are maintained as per the
specifications
• Use silk thread or single nylon fiber, flags or thin film tapes for
visualizing the air flow
• Place the tracer at the appropriate position such as at the
downstream of supply air and the return air risers as well as at
the doors opening, and check for the indication of the airflow
direction.
• Record the airflow pattern using still photography/videography.

Procedure (tracer particles)

• Ensure that the pressure diffentials are maintained as per the
specifications
• Generate the tracer particles by fogger/titanium tetrachloride or
equivalent.
• Position the tracer at the appropriate place, such as at the
downstream of supply air and the return air risers as well as at
the doors opening, and check for the indication of the airflow
direction.
• Record the airflow pattern using still photography/videography.
Air Flow Visualization (Unidirectional flow)
Objective
o To demonstrate unidirectional airflow and sweeping action over and away from
the product under dynamic conditions
Scope
o This test is applicable to those HVAC installations where the airflow direction is
unidirectional
Principle
o The flow visualization shall be confirmed by tracer particle generator (Fogger /
Titanium tetrachloride fumes shall be used for depicting the air flow pattern). The
profiles are recorded by storage devices such as video cameras, tapes or disks

Test Equipments
• Fogger or Titanium tetrachloride or equivalent
• Imaging equipment (Video recording or camera)

Procedure (tracer particles)

• Generate the tracer particles by fogger, titanium tetrachloride or
equivalent.
• Position the tracer at the appropriate place, such as at the
downstream of supply air and check for the indication of the
airflow direction.
• Record the airflow pattern using videography.
 Airborne Particle Count
Objective
o To establish that the critical locations within clean rooms comply with the air
borne particle count requirements as per specified for 0.5 and 5.0 µm particle size.
Scope
o This test is applicable to those HVAC installations where the cleanliness
classification is specified
Principle
o A Discrete Particle Counting (DPC), light scattering instrument shall be used to
determine the concentration of airborne particles, for particles equal to/larger than the
specified, at designated sampling locations and at specified occupancy states such as “At rest”
or “Operational”.
o At rest: Condition where the installation is complete with equipment installed
and operating in the manner agreed upon by the customer and supplier, but with no personnel
present.

• The number of sampling point locations by using the equation -

 NL = √A
 Where NL is the minimum number of sampling locations (rounded up
to next whole number)

Test Equipment
o Optical Particle Counter
o Volume of sample – 1 m3 equivalent to 35.3 ft3
Procedure

• Ensure that the area is tested for velocity, air changes,

differential pressure, installed filter leakages and is in
accordance with the respective test requirements.
• Set up the particle counter in accordance with the manufacturer’s
instructions and in compliance with the instrument calibration
certificate.
• Particle counting shall be done in the center of each room.
• Ensure that the sampling locations are positioned at the height of
the working activity and pointing into the airflow
• Ensure that the probe is positioned facing upwards parallel to the
flow (vertically or horizontally) with the opening towards the
HEPA filter (isoaxial sampling) at working height or 1m from the
floor.
• The volume sampled at each location shall be 1m3.
• Take count at each location as per the locations depicted in the
annexure 2.
• Air borne particle counting shall be done ‘at rest’ / static state.
Recovery/Decontamination rate test
Objective
o To determine capabilities of the system to recover from artificially generated
contamination. This test is to determine whether the clean room or clean zone is
capable of returning to its specified cleanliness class within a finite, predictable
time after being exposed briefly to an artificially induced excursion beyond
prescribed limit.
Scope
o The test is applicable to only non-unidirectional airflow systems in aseptic
process area and shall be carried at ‘at rest’ state of occupancy.
Principle
o Recovery performance is evaluated using the rate of change of particle
concentration
Test Equipment
o PAO/DOP Aerosol generator or equivalent
o DPC
Acceptance Criteria
o The overall recovery should take place within 10min.

Precaution
If the challenge particle concentration exceeds the maximum concentration capability of
the DPC used for the test, reduce the concentration to that level and determine the
recovery time from that concentration level.
Procedure
• Place the DPC probe at the testing point. The measuring points
and the number of measurements shall be as per annexure.
• The DPC probe shall not be placed directly under the supply
stream.
• Take the particle count in the area before aerosol generation at
‘at rest condition’. The sampling rate shall be 1 CFM.
• The particle size used in this test method shall be similar to that
during particle monitoring of minimum size.
• Artificially generate DOP/PAO aerosol in the classified area.
• With smoke generation output tube located at a pre-designated
location, generate smoke and check the count (1000 times more
then classified area at ‘at rest’). Record the particle count and
time.
• Stop the aerosol generator. The time at which the aerosol
generator is stopped shall be the starting time for establishing
the recovery rate or decontamination rate.
• Start the particle counting at the specified location at a sampling
rate of 1 CFM. Between two successive samples an interval of 2
 seconds shall be given. Establish the time required for attaining
the ‘at rest’ condition (denoted at Trn)
• Repeat for all other identified locations and record the recovery
time for each location as per annexure VII.
• Calculate the overall recovery rate of the room/area using
following equation.
N
• Tr all =
[∑1 Tr1 +1 Tr 2 +.......... .1 Tr n ]
o N = number of tested location.
o Tr all = is the recovery time of whole zone.
o Tr1= is the recovery time at each location.
Environmental Conditions – Temperature
Objective
o To demonstrate the ability of HVAC system to control temperature within a
prescribed level
Scope
o This test is applicable to those HVAC installations where the temperature
requirement is specified
Principle
oReadings of temperature shall be taken on hourly basis by using calibrated
instrument
Test Equipments
o Calibrated dry bulb thermometer, Thermo anemometer, automatic Temperature/
Humidity Recorder Hygrometers

Procedure
• Start the HVAC and allow the conditions to stabilize.
• Select the temperature monitoring location(s) (at least one) at
working height in each temperature –controlled zone.
• Select the Location for monitoring as per the following rationale:
• Near return air riser
• Near doors
• Near equipment/operation
• Allow sufficient time for sensor to stabilize the temperature
reading.
• Reading shall be taken for 3 consecutive days (minimum 16
hours/day) at 1 hour interval.
Environmental Conditions – Relative Humidity
Objective
o To demonstrate the ability of HVAC system to control relative humidity within a
prescribed level
Scope
o This test is applicable to those HVAC installations where the requirement of
relative humidity is specified
Principle
oReadings of Relative Humidity shall be taken on hourly basis by using calibrated
instrument
Test Equipments
o Calibrated dry bulb thermometer, Thermo anemometer, automatic Temperature/
Humidity Recorder Hygrometers

Procedure
• The test shall be done in conjunction with temperature test.
• Start the HVAC and allow the conditions to stabilize.
• Select the Relative Humidity monitoring location(s) (at least one)
at working height in each Relative Humidity controlled zone.
• Select the Location for monitoring as per the following rationale:
• Near return air riser
• Near doors
• Near equipment/operation
• Allow sufficient time for sensor to stabilize the Relative Humidity
reading.
• Reading shall be taken for 3 consecutive days (minimum 16
hours/day) at 1 hour interval.
Viable Count monitoring
Objective
o To demonstrate the ability of HVAC to control microbiological count below a
prescribed level
Scope
o This test is applicable to those HVAC installations where microbial limits are
specified and are critical for maintaining the respective cleanliness class
Principle
o Microbial load in the room shall be monitored through exposure of settle plates
and air sampling
• The number of sampling point locations by using the equation -
 NL = √A
 Where NL is the minimum number of sampling locations (rounded up
to next whole number)
Test Equipment
 Settle Plates.
 Centrifugal Air sampler
Procedure
• Ensure that the HVAC system is operational prior to test.
• Carry out sampling (settle plate and air sampling) at the
predetermined locations.
• Settled plates shall be of 90mm diameter and exposed for
duration of 1shift for grade A and 4hrs for grade B/C/D.
• Plates shall be exposed at a height above 1 meter from the floor
and also at work level for better exposure.
• Two plates (SCDA for TBC and SCA for TFC) shall be exposed
at all the specified locations.
• For air sampling, 1m3 of air from specified locations shall be
sampled using Soybean Dextrose Agar.
• Incubate settle plate at 20 – 25 °C (for TFC) for five days and at
30 – 35°C (for TBC) for three days.
• Incubate the air sampling Petri plates at 30 – 35°C for three days
• Perform the test for 3 consecutive days.

o Surface Monitoring: Floor, walls and equipments shall be monitored using swab
technique or 25 cm2 contact plates. Select the particular location of the
equipment / floor / wall, take out the swab from the test tube, apply the cotton
swab left hand side to right hand side horizontally to cover an area of 5 x 5 cm.
Replace the swab immediately inside the swab holder test tube Perform
microbiological testing. Follow pour plate method for determining the bacterial or
fungal organisms using Soybean Casein Digest Agar. Incubate all the agar plates
for 72 hours for bacterial growth between 20-25 deg.C and further for 48 hours
for fungal growth at 30-35 deg.C. After incubation observe the plates for any
bacterial or fungal growth and multiply with dilution factor and report the results.
o Active Air Sampling: For air sampling, 1m3 of air from specified locations shall
be sampled using Pre-sterilized Soybean Casein Agar plates. Plates shall be of
90 mm diameter. Incubate all the agar plates for 72 hours for bacterial growth
between 20-25 deg.C and further for 48 hours for fungal growth at 30-35 deg.C.
After incubation observe the plates for any bacterial or fungal growth and report
the results.
 o Passive Air Sampling: Carry out sampling (90 mm diameter plate) using pre-
sterilized Soybean Casein Agar plates at the pre-determined locations. Plates
shall be exposed for four hours at a height above 1 meter from the floor and also
at work level for better exposure. Incubate all the agar plates for 72 hours for
bacterial growth between 20-25deg.C and further for 48 hours for fungal growth
at 30-35 deg.C. After incubation
 observe the plates for any bacterial or fungal growth.
 Dehydrated media make – HIMEDIA or DIFCO shall be used. The
required quantity of dehydrated media shall be sterilized as per the
current document on media preparation. Keep inoculated and un-
inoculated agar plates as positive and negative control to confirm validity
or results. Positive control plates must show growth (indicating growth
promotability of the medium), whereas negative control plates must not
show any growth (indicating sterility of the medium). Soybean Casein
Digest Agar to be tested for Growth Promotion Test (GPT), positive
control shall be checked by inoculating between 10 to 100 cells of
Bacillus subtilis and Candida albicans separately. Reject all the counts if
any growth is obtained in negative control

 Surface Monitoring: Floor, walls and equipments shall be monitored

using swab technique or 25 cm2 contact plates.
 Select the particular location of the equipment / floor / wall, take out the
swab from the test tube, apply the cotton swab left hand side to right
hand side horizontally to cover an area of 5 x 5 cm.
 Replace the swab immediately inside the swab holder test tube Perform
microbiological testing. Follow pour plate method for determining the
bacterial or fungal organisms using Soybean Casein Digest Agar.
 Incubate all the agar plates for 72 hours for bacterial growth between 20-
25 deg.C and further for 48 hours for fungal growth at 30-35 deg.C.
 After incubation observe the plates for any bacterial or fungal growth and
multiply with dilution factor and report the results.
Re-qualification Criteria
o The periodic re-qualification shall be done at a time interval as specified in the
standard operating procedure.
o Other than periodic re-qualification, the HVAC shall be validated in following
cases:
 Any major breakdown
 Any modification in operation process
 Any change in test parameters
 All the above cases shall be evaluated for their impact on quality
parameters, and the test criteria shall be defined based on this
evaluation.