0 PERFORMANCE QUALIFICATION TEST

Following Performance Qualification studies shall be carried out to ensure the equipment for
proper operation and its ability to sterilize and depyrogenate the washed vials at the set
parameters, repeatedly & consistently.
Air Velocity measurement.
HEPA FILTER Integrity test by DOP / PAO Aerosol test.
Air flow pattern test.
Non-viable airborne particle count test.
Heat Distribution studies.
Heat Penetration studies.
Endotoxin Challenge study.

To qualify the test the equipment should fulfill the acceptance criteria described in the individual
test procedures. All instruments shall be calibrated before starting and after completion of
validation studies.
After completion of the qualification tests all the data generated will be compiled to evaluate
ability of the sterilization and depyrogenation tunnel to sterilize & depyrogenate different articles
at the set operating parameters.
2.0 TEST CALIBRATION EQUIPMENTS
S. No Equipment Equipment Calibration on Calibration Due on Verified By
Name I.D

2.1 QUALIFICATION OF THE EXECUTION TEAM:

The execution team should be trained for execution of operation Qualification of the equipment
& Record should be maintained in Annexure 1
2.2.1 AIR VELOCITY MEASUREMENT
2.2.1.1 OBJECTIVE
To demonstrate that the system is capable of delivering air velocities, as per the Requirement, to
maintain continuous laminarity of HEPA Filter installed in tunnel.
2.2.1.2 EQUIPMENT USED
Digital Anemometer
2.2.1.3 METHOD
A) This test shall be performed by a trained personal and training record shall be attached with
report.
B) This test shall be performed at least 30 minute after the tunnel has been Started Heater should
be off.
C) Measure Air velocity 6 inches below the filter at 5 locations (four corner filter and
one center), calculate the average velocity of air coming from supply.
D) Calculate the velocity of air coming from supply filter should be measured in Feet/min.
E) Record the Data in Annexure II.

V1 V2
 V3

V4 V5

Average velocity (A) = V1+V2+V3+V4+V5/5

A = Average
2.2.1.4 ACCEPTANCE CRITERIA
Average velocity across HEPA Filter should be within the range of 9020 Feet/min
Average velocity across HEPA Filter in sterilization zone should be within the range of 150-
200Feet/min
2.2.2 HEPA FILTERS LEAKAGE TEST (DOP/PAO TEST)
2.2.2.1 OBJECTIVE
To verify the integrity of HEPA FILTERS installed in the sterilization
and depyrogenation tunnel.
2.2.2.2 TEST APPARATUS
Aerosol Generator
Aerosol photometer
Compressed Air

2.2.2.3 METHOD
a) Filter testing shall be performed only after operational air velocities have been verified and
adjust wherever necessary.
b) Position the smoke generator and introduce (DOP/PAO) smoke into the stream of the HEPA
Filter. The concentration of (DOP/PAO) aerosol is 80-120g per liter of air.
c) Programme the instrument at 100% concentration for upstream.
d) Scan the downstream side of the HEPA with an appropriate photometer probe.
e) The probe shall scan the entire filter face and frame at a position about 1 inch from the face of
the filter.
f) Scanning shall be done at the rate of 2 feet/minute.
g) Record the observation in Annexure III
2.2.2.4 ACCEPTANCE CRITERIA
During scanning percentage of DOP or PAO penetration shown by photometer should not be
more than 0.01% of the upstream concentration through the filter media and should be zero
through mounting joints.
2.2.3 AIR FLOW LAMINARITY:
2.2.3.1 OBJECTIVE
To determine the air flow pattern of HEPA Filter installed in sterilization
and depyrogenation tunnel
2.2.3.2 EQUIPMENT USED
Titanium Tetra Chloride (Chemical Used).
Video Camera.
2.2.3.3 METHOD
TITANIUM TETRA CHLORIDE
a) Take the glass rod with cotton or sponge tied to it.
b) Dip the rod in Titanium Tetra Chloride solution and place the stick at Downstream of HEPA
Filter.
c) Check the air flow direction at the Downstream of the filter face.
d) Observe & record the same with video camera.
2.2.3.4 ACCEPTANCE CRITERIA
The stream of air should be unidirectional and non-turbulent upto working zone.
2.2.3.5 Record the result in Annexure V
2.2.4 AIR BORNE NON-VIABLE PARTICLE COUNT TEST
2.2.4.1 OBJECTIVE
To establish that at different locations within the tunnel, a count size of particles per cubic meter
is within the limit.
2.2.4.2 EQUIPMENT USED
Air Borne Particulate Counter, duly calibrated with traceability certificate.
2.2.4.3 METHOD
a) The test should be perform only when the earlier test have been performed (air velocity, filter
integrity, air laminarity) and found to be conforming to the requirement. The particle count test
should be performed by qualified and/ or trained personnel only.
b) Calculate the number of location by the following formula
No. of sampling location: NL=A
Where:
NL=The minimum number of sampling locations (Rounded up to a whole number)
A=The area of the HEPA Filter installed in tunnel in m2
c) Use calibrated particle analyzer for non viable particle count and consider 0.5 and 5.0
d) Perform 3 consecutive cycles.
e) Record the data in Annexure VIII
2.2.4.4 ACCEPTANCE CRITERIA
The system shall capable of achieving the desired cleanliness grade in the subjected critical work
locations in the tunnel as per the designed specified limit
CRITERIA ACCEPTANCE
Maximum number of permitted particles per cubic meter equal to or
above
Non viable Air particle
GRADE 0.5 m 5.0 m
count test
GRADE A 3520 29
2.2.5 HEAT DISTRIBUTION STUDY
2.2.5.1 OBJECTIVE
The objective of the test is to ensure that:
The Sterilizing & Depyrogenating Tunnel when operated with Empty Chamber is capable of
producing the temperature profiles as per the temperature set points set in the PLC of the
equipment. The temperature distribution is uniform throughout the sterilization period.
Three run to be performed to qualify the measurement of the temperature throughout the
chamber during the sterilization cycle.
2.2.5.2 Equipment Used
Data Logger with Minimum 12 Probes duly calibrated.
2.2.5.3 METHOD
Record the set parameters of the sterilization cycle to be operated during the test. Suspend the
probe in the chamber in different position in such a way that probes dont touch any metallic
surface. Record the position of the probe in a representative schematic manner.
Connect the probes to suitable data logger, which can scan and print the actual temperature
observed at different locations with respect to time.
Operate the Sterilizing & Depyrogenation Tunnel Also start the data logger to record the actual
temperatures with respect to time.
After completion of sterilization cycle switch off the data logger.
Collect print out from the printer of Sterilizing & Depyrogenation Tunnel.
Download the data from the data logger in the computer for the data analysis and printing
enclosed the printout obtained from the data logger.
2.2.5.4 ACCEPTANCE CRITERIA:
The Temperature at each Temperature probe should be 300C during the cycle.
2.2.5.5 Observations and Results In Annexure VI
Record the temperature at various locations
2.2.6 HEAT PENETRATION STUDY
2.2.6.1 OBJECTIVE
Objective of this test is to ensure that, heat is sufficiently penetrating into the innermost portion
of the Vial subjected for sterilization & Depyrogenation to achieve desired temperature during
the sterilization & Depyrogenation cycle.
Loaded chamber Heat penetration studies must be conducted for three consecutive cycles with
temperature probes.
The recovery of Endotoxin Concentration after exposing to Depyrogenation tunnel should show
more than 3 log reduction.
Three run to be performed to qualify the measurement of the temperature throughout the tunnel
by seven (during the sterilization cycle).
2.2.6.2 EQUIPMENT USED
Data Logger with 12 Probes duly calibrated.
2.2.6.3 METHOD
Conduct the study with loaded chamber for three consecutive cycles with temperature probes and
Endotoxin vials.
Suspend the 12 probes inside the vial and put into tunnel for Heat Penetration Study.
Record the position of the probes in a representative schematic form.
Insert 9 Endotoxin Vials (Marked vials) having 10000 EU each along with the temperature
sensors.
Connect the probes to suitable data logger, which can scan and print the actual temperature
observed at different locations with respect to time.
Operate the Sterilizing & Depyrogenating Tunnel also start the data logger to record the actual
temperatures within the Sterilizing & Depyrogenating Tunnel with respect to time.
BELT SPEED CAN BE CALCULATED BY USING THE FOLLOWING FORMULA:

(Vial diameter)2 cos30o washing machine output

Tunnel conveyor width

STERILIZATION HOLD TIME: Length of sterilizing zone

Conveyor speed
2.2.6.4 ACCEPTANCE CRITERIA
Throughout the dwell time, all temperature measured in the chamber is 300C. The recovery
of endotoxin concentration after exposing in sterilization and depyrogenation should show at
least 3 log reduction.
3.0 ACCEPTANCE CRITERIA
S.No. TEST PERFORMED ACCEPTANCE CRITERIA
1. Air velocity test Average velocity across the HEPA filter should be within
the range of 90 20% FPM.
2. HEPA Filter integrity Leakage in the filter should not be more than 0.01% and
testing should be zero through mounting joints.
3. Air flow pattern Air flow pattern should be unidirectional and non turbulent.
4. Non Viable particle Acceptable limit of
counter 0.5 Particles is 3520 and 5.0 is 29 in 1M3 of air.
5. Heat distribution studies Temperature should be uniform throughout the cycle
6. Heat penetration studies Should show at least 3 log reduction after exposing
Sterilization temperature
4.0 RESULT:
The results obtained after execution of the protocol, will be presented in the validation
report. All results will be compared against the acceptance criteria. Any deviation failure to
meet these specifications will be duly documented. A deviation indicates variance from the
acceptance criteria however, does not necessarily mean that the qualification as unacceptable.
During execution of this protocol, if any deviation is noticed, the person executing the protocol
initiates a deviation report and provides the detail description of the deviation. Investigation may
be performed to identify root cause for the deviations or failures and corrective action will be
taken to minimize such deviations or failures in the future.
5.0 CONCLUSION:
Conclusion will be drawn after compilation and evaluation of result.

Related: How to Write a Validation Protocol?

ANNEXURE 1
Training for Performance Qualification of sterilization and depyrogenating Tunnel
Name of the trainer :
Designation :
Date of training :
Duration :
Sr. No. Name of Trainee Department Signature

Comments _____________________________________________________________
Training given by _______________ _______________
Name Sign/Date

ANNEXURE II
AIR VELOCITY MEASUREMENT TEST
Date of Test : __________________
Equipment ID:
Name of testing instrument: Digital Anemometer
Calibrated on : ____________________ Calibration due: ___________

HEPA Velocity [FPM] Average

FILTER Velocity
Location Location 2 Location 3 Location 4 Location 5
No. [FPM]
1 (V1) (V2) ( V3) (V4) (V5)
1

Checked by
(Production) _________________ _________________ ____________
(Name) (Signature) (Date)
Verified by
(QA) _________________ _________________ ____________
(Name) (Signature) (Date)

ANNEXURE III
HEPA FILTER INTEGRITY TESTING
Date of Test : ________________
Equipment ID :
Name of Testing Instrument: Aerosol Photometer
Calibrated on : ________________ Calibration due: _______________

HEPA Filter No. Upstream Downstream Remarks

Concentration Concentration
%age %age (NMT 0.01%)
 1 100%
2 100%
3 100%
4 100%
5 100%
6 100%
Checked by
(Production) _________________ _________________ ____________
(Name) (Signature) (Date)
Verified by
(QA) _________________ _________________ ____________
(Name) (Signature) (Date)

ANNEXURE IV
AIR FLOW PATTERN TEST

Date of Test :
Equipment ID :
Teat Conditions : Static
Name of the Testing Chemical : Titanium Tetra Chloride
CONDITION CRITERIA YES/NO
STATIC Whether the fumes of the titanium
tetra chloride follow unidirectional
path

Checked by
(Production) _________________ _________________ ____________
(Name) (Signature) (Date)
Verified by
(QA) _________________ _________________ ____________
(Name) (Signature) (Date)

ANNEXURE V
NON VIABLE PARTICLE COUNT TEST

Date of Test : ____________ Equipment ID:

HEPA Filter No :
Test Conditions : Static
Name of Instrument : Air Borne Particulate Counter
Calibrated on : ________________
Calibration due : ________________
Location Particle Particulate Count Average Average
 No. Size Count Count Count3 Count Count (A=C1 In M3
1 2 (C3) 4 5 +C2 +-- (Am =A*
(C1) (C2) (C4) (C5) +Cn )/n 35.2)
L1 0.5
5.0
L2 0.5
5.0
L3 0.5
5.0
L4 0.5
5.0
L5 0.5
5.0

ANNEXURE VI
HEAT DISTRIBUTION STUDY
TEST INSTRUMENT DETAILS TO BE USED FOR VALIDATION
Test Model no Calibration done
instrument date
name
Sensor type Make Calibration due
& quantity date

CYCLE NO
Equipment Date of Equipment id no
name experiment
Equipment Cycle start Cycle end
make
Equipment
location

STERILIZATION CYCLE PARAMETERS

Conveyor Sterilization Hold time

Start temperature
 Conveyor Belt speed in mm
Stop temperature
Sterilization
temperature

STERILIZATION CYCLE OBSERVATION

TIME TEMPERATURE SENSORS
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 MEAN
Ple. Refer temp mapping
date and print out of
reading & temperature
mapping data as attachment

Inference __________________________________________________
__________________________________________________

Reviewed By Reviewed By Approved By Head

Engineering Head Production Head -QA

ANNEXURE VII
HEAT PENETRATION STUDY
TEST INSTRUMENT DETAILS TO BE USED FOR VALIDATION
Test instrument Model no Calibration done
name date
Sensor type Make Calibration due
& quantity date

CYCLE NO
Equipment Date of Equipment id no
name experiment
Equipment Cycle start Cycle end
make
Equipment
location

STERILIZATION CYCLE PARAMETERS

Conveyor Sterilization Hold -----
Start time
temperature
Conveyor Belt speed in -----
Stop mm
temperature
Sterilization ----- -----
temperature

STERILIZATION CYCLE OBSERVATION

TIME TEMPERATURE SENSORS
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 MEAN
Ple. Refer temp mapping
date and print out of
reading & temperature
mapping data as attachment

Inference___________________________________________________________________
______________________________________________________________________________
________
Reviewed By Reviewed By Approved By
Head Engineering Head Production Head -QA