Cleanroom Validation

HMA MEDICAL LTD
Plot 9, Block 4, Transit Camp Road, Ilorin, Kwara State, Nigeria.

Title: PERFORMANCE QUALIFICATION PROTOCOL FOR EXTENDED CLEANROOM
QM No: HMA/QAD/CVP/001-01 Effective Date: 9th January, 2024
SUPERSEED: HMA/QAD/CVP/001-00 Next review Date: 8th January, 2026
PERFORMANCE
QUALIFICATION PROTOCOL
FOR CLEANROOM
TABLE OF CONTENT
1. Protocol Approval sheet 3
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HMA MEDICAL LTD
2. Objectives 4
3. Scope 4
4. Responsibility 4
5. General Definitions 4
6. Equipment and instruments 5
7. Test Procedures 5
7.1 Differential Pressure Test 5
7.2 Temperature and Humidity Control Test 6
7.3 Illumination Level 7
7.4 Noise Level 8
7.5 Non-Viable particulates count 9
7.6 Viable particulates count(surface) 10
7.7 Viable particulates count(air) 11
7.8 Air Velocity test and air changes per Hour 12
7.9 Recovery Test 13
8. Abbreviations 14
9. Reference 15
10. Annexures 16
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HMA MEDICAL LTD
1. Protocol Approval sheet

This protocol shall be prepared, reviewed, approved and authorized by underlisted
Name Designation Signature Date

Prepared by Quality Assurance Executive
Reviewed by Quality Control Manager
Approved by Quality Assurance Manager
Authorized by General Manager
2. OBJECTIVES
The objective of this protocol is to establish a systematic approach to validate that the
environment of the production area in which the product and components shall be
exposed to during its manufacturing process is suitable and appropriate under
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HMA MEDICAL LTD
acceptable conditions and parameters of the design in compliance with the

International Organization ISO under standard ISO 14644.
3. SCOPE
This protocol shall be applicable to the validation process for class C Cleanroom (ISO
7 Cleanroom) used in the manufacturing of Deleject syringe and needles and its
components.
4. RESPONSIBILITY
4.1 QC Microbiologist: responsible to arrange the sampling and testing of the
validation samples.
4.2 QA Executive: shall prepare the validation protocol as per the guideline of
validation master plan and conduct the validation compilation and summary
preparation.
4.3 Quality Control Manager: shall check the summary and results of the
executed protocol for compliance and reporting any discrepancies found for
necessary corrective and preventive actions.
4.4 Quality Assurance Manager: Responsible for the final review and
approval of protocol and report.
5. GENERAL DEFINITIONS
5.1 ISO 7 cleanroom (Class 10,000 cleanroom) is a hard sided wall
manufactured facility that utilizes HEPA filtration systems to maintain air
cleanliness levels of a maximum of 10,000 particles (≥ 0.5µm) per cubic foot.
5.2 Cleanroom: a room within which the number concentration of airborne
particles is controlled and classified, and which is designed, constructed and
operated in a manner to control introduction, generation and retention of
particles
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HMA MEDICAL LTD
inside the room.

5.3 Viable Particulates: these are small size living organisms such as bacteria,
fungi, mould, algae, etc.
5.4 Non-Viable Particulates: this are formed by disintegration of large size
materials or condensation of small size particles or droplets, e.g., mist, smoke,
fume and dust.
6. EQUIPMENT AND INSTRUMENT

6.1 Anemometer
6.3 Temperature and Humidity Meter
6.4 Light level meter-lux meter
6.5 Noise level meter-dBA meter
6.6 Air particulate counter
6.7 Air sampler
6.8 Fog (Smoke) generator
6.9 HVAC Meter
7. TEST PROCEDURE
7.1 Differential Pressure Test
This test is done to demonstrate the capability of the air system to provide pressure
gradient among the different room
7.1.1 Acceptance criteria
CRITERIA ACCEPTANCE
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HMA MEDICAL LTD
Differential pressure of rooms Between Different Classes of Clean Room

(Area) ≥ 5Pa
Between Clean room (Area) and non-Clean
room (Area) ≥10Pa
Between Clean room(Area) and Outside
barometric pressure ≥10Pa
7.1.2 Procedure:
STEP ACTION
1. Enter in the respective area and ensure that all doors are tightly closed and HVAC
in running condition before taking the pressure reading.
2. To avoid unexpected changes in pressure and to establish baseline, all doors in
sterile areas must be closed and no traffic is to be allowed through the facility
during test.
3. Take the magnehelic reading in the reference of the negative and positive
pressure of rooms
4. Record the pressure displayed on the magnehelic guage three times as per
Annexure 1
7.2 Temperature and Humidity Control Test
This test is performed to demonstrate the ability of the Air Handling Unit to provide
the required temperature and control relative humidity at specified level in critical and
controlled areas. The two parameters are monitored in continuous manner,
documented in the format and stabilization is ensured within the specified limit.
7.2.1 Acceptance Criteria
CRITERIA ACCEPTANCE
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HMA MEDICAL LTD
Temperature in controlled areas ≤ 25oC

Temperature in critical areas ≤ 28oC
Humidity in controlled areas 45% ±5%
Humidity in critical areas NMT 55%
7.2.2 Procedure:
STEPS ACTIONS
1.5.1.1.1 Hang the Temperature and Humidity meter on the wall
2.5.1.1.2 Stretch the wire probe out in other to get accurate values.
3.5.1.1.3 Temperature and Humidity meter should not be keep under direct airflow and in front of the
window where direct sun light exposed.
4.5.1.1.4 Ensure a calibrated Temperature and Humidity meter is used
5.5.1.1.5 Enter in the respective locations and take the reading.
6.5.1.1.6 Check the temperature and relative Humidity display on Temperature and Humidity meter
7.5.1.1.7 Record the value in the chart (Annexure 2)
7.3 Illumination Level
The objective of the test is to determine the lightning level and its uniformity
throughout an area at predetermined point.
CRITERIA ACCEPTANCE
Light level Clean Room (Area) ≥ 300Lux
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HMA MEDICAL LTD
Supplementary area ≥ 200Lux
7.3.2 Procedure
STEPS ACTION
1. Insert the prescribed battery into the digital lux meter to power it on.
2. Put on the power switch to activate the tester.
3. Then open the lid covering the photoreceptor to expose it to light for which its
intensity is to be determined.
4. Extract the readings from the instrument for record purpose. (Annexure 3)
5. Note down all variations from the specification if any
7.4 Noise Level
This test is to determine the noise level and its uniformity throughout an area at
predetermined points.
CRITERIA ACCEPTANCE
Noise Level ≤ 65dB
7.4.2 Procedure
STEPS PROCEDURE
1. Verify that the HVAC is operating in a stable condition and all equipment that is
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HMA MEDICAL LTD
specified is in operational condition.

Set the instrument to work and ensure unauthorized entry and exits from the
room is prevented
Measure the sound levels at work surface height or 750mm from the finished
floor level and layout of noise level.
Record the data as per Annexure 4
Note down all variations from the specification if any.
7.5 NON-VIABLE PARTICUATE COUNT
The objective of this test is to establish that areas meets predefined conditions for
non-viable particles. Particle counter is used to conduct the test. Particle count is
taken at static conditions before the operations as well as operational working
condition. The particle count should be within the range as per the standards of
particle classification.
CRITERIA ACCEPTANCE
Maximum number of permitted particles per cubic meter equal to or above

Grade AT REST IN OPERATION
Non-viable ≥0.3µm ≥0.5µm ≥1.0µm ≥5.0µm ≥10.0µm ≥0.3µm ≥0.5µm ≥1.0µm ≥5.0µm ≥10.0µm
particulate A 10200 3520 832 29 1 10200 3520 832 29 1
count test B 102000 35200 8320 293 2000 102000 35200 8320 293 2000
C 352000 83200 2930 20000 352000 83200 2930 20000
D 3520000 832000 29300 3520000 832000 29300
7.5.2 Procedure
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HMA MEDICAL LTD
STEP PROCEDURE
S
1. Calculate the number of sampling points with the formula N= square root of the volume
of the room in cubic meter, make it approximately. Reference is made to ISO14644-1
2. The sampling location shall be evenly distributed throughout the area of clean room or
clean zone and counter is positioned at the height of work activity.
3. Operate the air particle counter as per SOP HMA/QAD/SOP031-04
4. Record the data as per Annexure 5
5. Note down all variations from the specification if any.
7.6 VIABLE PARTICULATE COUNT (SURFACE)
Viable particle count is performed on weekly basis by employing the swab test and
using nutrient agar medium for the incubation of microorganisms.
CRITERIA ACCEPTANCE
Floor swab
Bacteria ≤ 25 CFU
Fungi ≤ 5 CFU
7.6.2 Procedure
STEPS PROCEDURE
1. Verify that the HVAC is operating in a stable condition
2. Collect swab sample from the surface and analyze without delay.
3. Take microbial count of the sample as per SOP HMA/QAD/SOP042-02
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HMA MEDICAL LTD
7.7 VIABLE PARTICULATE COUNT (AIR)
The objective of the test is to determine the level of atmospheric microorganisms in

clean areas. The different media plates are exposed in every manufacturing section.
The microorganism count should be within the range, otherwise an investigation must
be initiated to evaluate the root cause, effective corrective and preventive action
CRITERIA ACCEPTANCE
Air
Bacteria ≤ 50 CFU
Fungi ≤ 50 CFU
7.7.2 Procedure
STEPS ACTION
1. Verify that the HVAC is operating in a stable condition
2. Sterile nutrient agar and Sabouraud dextrose agar should be poured aseptically
into different sterile 90mm X 90mm by 4inches Petri-dishes and allowed to
solidify.
3. Plates should be labeled accordingly
4. Expose each of the plates containing the solidified agar at different locations of
the non-clean and clean Area for at least four hours.
5. The plates containing Nutrient agar should be incubated upside down in an
incubator at 37°C for 24- 48 hours while that containing Sabouraud dextrose agar
should be incubated at 25°C for 72 hours
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HMA MEDICAL LTD
7.8 AIR VELOCITY TEST AND AIR CHANGES PER HOUR
The objective of the test is to ensure that the required air is delivered by HEPA filters
and to calculate the air changes per hour of each area.
CRITERIA ACCEPTANCE
Air Changes Per Hour 20-40 unit/h
7.8.2 Procedure
STEP ACTION
1. Scan the area of the HEPA Filter, with the anemometer probe, 6 inches from the
filter face.
2. Divide the area of the HEPA Filter into four equal, hypothetical grids.
3. Record the velocity reading taken at the center of the grids, and at the junction of
dividing lines (center of HEPA Filter)
4. Calculate the average velocity of each filter
7.8.3 Calculation of Air Changes per Hour

STEP ACTION
1. Calculate the actual volume of supplied air in each room as below
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HMA MEDICAL LTD
2. Air flow discharge [m3/minute] = Air Velocity of HEPA filter

3. Calculate the volume of the room in cubic meter, of which air changes are to be
calculated
4. NO of ACPH = Airflow discharge in room X 60(CMH) / room volume
5. For all classified areas, ACPH should not be less than 20
6. If the air changes in the area are not within the limit, trouble shoot the cause
and rectify
7. Record the ACPH as in Annexure 8
8. Refer to SOP HMA/QAD/SOP055-02
7.9 RECOVERY TEST
This test is performed to determine how much time it takes to recover the expected
conditions. The recovery of temperature and humidity conditions is checked after
losing operational power conditions or door opening. The time required to stabilize
the temperature and humidity is noted.
CRITERIA ACCEPTANCE
Recovery Time 10minutes
7.9.2 Procedure
STEP ACTION
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HMA MEDICAL LTD
1. Note the temperature and the humidity of the area under test
2. Shut off the HVAC system
3. Wait for the Temperature and Humidity of the area under test to stabilize
4. Operate HVAC system and note the time
5. Wait to stabilize the Temperature and Humidity in the area within the specification
limit
6. Note and record the time
8. ABBREVIATIONS
8.1 AHU – Air Handling Unit
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8.2 NMT – Not More Than

8.3 CFU – Colony Forming Unit
8.4 DB – Decibel
8.5 HVAC – Heating Ventilation and Air Conditioning
8.6 HEPA – High Efficiency Particulate Air Filter
8.7 ACPH – Air changes Per Hour
9. REFERENCES
ISO 14644
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HMA MEDICAL LTD
SOP: HMA/QAD/SOP010-02
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10 ANNEXURES
ANNEXURE 1
DIFFERENTIAL PRESSURE TEST RESULT
Pressure Differential
(Pa)
No. Clean Room Remark
1st 2nd rd
3
Readin Readin
Reading
g g
1.
2.
3.
4.
5.
Between Different Classes of Clean Room (Area) ≥ 5Pa

Standard Between Clean room (Area) and non-Clean room (Area) ≥10Pa
Between Clean room(Area) and Outside barometric pressure ≥10Pa
Conclusion
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ANNEXURE 2
TEMPERATURE AND HUMIDITY CHART
MONTH …………………………...……………….
LOCATION:
Temperature (T oC) & Relative Humidity (H%)
Actual 1st Reading Actual 2nd Reading Actual 3rd Reading
Date Time (7:00AM-9:00AM) Time (12:00-2:00PM) Time (5:00PM-7:00PM) Mean Value
Taken Taken Taken Done By Remark
AM o
T( C) H(%) PM o
T( C) H(%) PM o
T( C) H(%) o
T( C) H(%)
Standard Temperature: ≤ 25℃ Humidity: 45%±5%

Conclusion
Remark:
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ANNEXURE 3
ILLUMINATION LEVEL TEST RESULT
Test Date:
Illuminance (Lux)
No
Test site 1st 2nd 3rd Remark
. Mean
Readin Readi Readi
Value
g ng ng
1.
2.
3.
4.
5.
Clean Room (Area) ≥ 300Lux

Standard
Supplementary area ≥ 200Lux
Conclusion
Remark:
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ANNEXURE 4
NOISE LEVEL TEST RESULT
N Noise Index
o. Clean Room (dB) Remark
1.
2.
3.
4.
5.
Standar
≤ 65dB
d
Conclusi
on
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ANNEXURE 5
AIR PARTICLE COUNT RECORD SHEET
Site: Date:
Maximum Concentration Limits (particles/m3 of Air) for particles ≥ Size Zones
Gra At Rest
de ≥ 0.3µm ≥ 0.5µm ≥ 1.0µm ≥ 3.0µm ≥ 5.0µm ≥ 10.0µm
A 10200 3520 832 29 1

B 10200 35200 8320 293 2000
0
C 35200 83200 2930 20000
0
D 35200 83200 29300
00 0
Gra In Operation
de ≥ 0.3µm ≥ 0.5µm ≥ 1.0µm ≥ 3.0µm ≥ 5.0µm ≥ 10.0µm
A 10200 3520 832 29
B 10200 35200 8320 293
0
C 35200 83200 2930
0
D 35200 83200 29300
00 0
Remark:
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ANNEXURE 6
VIABLE PARTICULATE COUNT (Surface-Floor swab)
NA: 370C/48hrs
INCUBATION
SDA: 220C-250C/120hrs
Colonies Count/Hour
Analysis Area Spec 24hrs 48hrs 72hrs 96hrs 120hrs Result Done Checked
S/N Location Classification Sample Date By By
Remark
Date CFU
B F B F F F F
FS
FS
FS
FS
FS
FS
FS
FS
FS
FS
FS
FS
FS
FS
Key Limit: -
FS – Floor Swab NA – Nutrient Agar Alert Limit:
B – Bacteria SDA – Sabouraud Dextrose Agar Action Limit:
F – Fungi
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ANNEXURE 7
VIABLE PARTICULATE COUNT(Air)
Date of Plate Exposure: - Time of Exposure
From………. To……….
Pre-fumigation/Post fumigation Bioburden Active/Passive Bioburden
NA- Incubated @370C/48hrs
SDA – Incubated @250C/120hrs
Colonies Count/Hour
Analysis Area Spec Done Checked
S/N Location Classification 24hrs 48hrs 72hrs 96hrs 120hrs Result Date
By By
Remark
Date CFU
B F B F F F F
1.
2.
3.
4.
5.
Key
B – Bacteria NA – Nutrient Agar
F – Fungi SDA – Sabouraud Dextrose Agar
CFU – Colony Forming Unit NC- Non-Classified
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ANNEXURE 8
AIR VELOCITY AND AIR CHANGES PER HOUR
DATE: QA Form-014
No Description of Location Dimension(m) Area Volume Total air volume Air Changes ACPH AHU Class Temp
Delivered/time per hour Specs Capacity
(CADR) (ACPH)
L W H m2 m3 M3p/hr. Ft3/min Unit/h o
C
1.
2.
3.
4.
5.
6.
24

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HMA MEDICAL LTD

Plot 9, Block 4, Transit Camp Road, Ilorin, Kwara State, Nigeria.

1. Protocol Approval sheet

Name Designation Signature Date

acceptable conditions and parameters of the design in compliance with the

inside the room.

6. EQUIPMENT AND INSTRUMENT

7.1 Differential Pressure Test

7.1.1 Acceptance criteria

Differential pressure of rooms Between Different Classes of Clean Room

7.2 Temperature and Humidity Control Test

7.2.1 Acceptance Criteria

Temperature in controlled areas ≤ 25oC

7.3 Illumination Level

7.3.1 Acceptance Criteria

Supplementary area ≥ 200Lux

7.4 Noise Level

7.4.1 Acceptance Criteria

specified is in operational condition.

7.5 NON-VIABLE PARTICUATE COUNT

7.5.1 Acceptance Criteria

Maximum number of permitted particles per cubic meter equal to or above

7.6 VIABLE PARTICULATE COUNT (SURFACE)

7.6.1 Acceptance Criteria

7.7 VIABLE PARTICULATE COUNT (AIR)

The objective of the test is to determine the level of atmospheric microorganisms in

7.7.1 Acceptance Criteria

6. Record the data as per Annexure 7

7. Note down all variations from the specification if any.

7.8 AIR VELOCITY TEST AND AIR CHANGES PER HOUR

7.8.1 Acceptance Criteria

7.8.3 Calculation of Air Changes per Hour

1. Calculate the actual volume of supplied air in each room as below

2. Air flow discharge [m3/minute] = Air Velocity of HEPA filter

7.9 RECOVERY TEST

7.9.1 Acceptance Criteria

2. Shut off the HVAC system

4. Operate HVAC system and note the time

8.2 NMT – Not More Than

Between Different Classes of Clean Room (Area) ≥ 5Pa

Standard Temperature: ≤ 25℃ Humidity: 45%±5%

Clean Room (Area) ≥ 300Lux

A 10200 3520 832 29 1

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