Autoclave Validation Protocol

OBJECTIVE:
The purpose of this document is to provide the documented evidences which prove that, the performance
of Sterilizer with Bung Processor is in accordance to the specifications and meet the requirements of the
product, process, safety, regulatory bodies and cGMP obligations.
SCOPE:
The scope of this Re-qualification exercise is verification and compilation of Re-qualification study conducted
and documentation on this study.
EQUIPMENT DESCRIPTION:
STERILIZER WITH BUNG PROCESSOR
Identification Number ---
Location ---
Purpose To sterilize the bungs, Gowns, Tools and equipment parts.
QUALIFICATION TEAM:
Supervisor Engineering Department.
Chemist Production Department.
Microbiologist.
Q.A Representative.
Authorized personnel of External Agency.
RESPONSIBILITIES:
Supervisor Engineering Department.
Co-ordinate with External Agency for conducting validation of Sterilizer
with Bung Processor.
Chemist Production Department. Machine operation and execution of the protocol.
Authorized personnel from External
Agency.
To conduct heat distribution and penetration studies.
Microbiologist. To conduct the Biological indicator test.
Q.A Representative.
Preparation of the protocol.
Execution of the protocol.
To verify and document the results gathered during the validation of
Sterilizer with bung processor.

IDENTIFICATION OF CRITICAL CONTROL MONITORING PARAMETER:
Before proceeding for Qualification of Autoclave with bung processor, following parameters to be checked:
Biological Indicator spore concentration must be 106.
Used media must be tested for its growth promoting test.
Calibration of Pressure Gauge
Calibration of Temperature Gauge
Calibration of PT-100 Sensors
RE-QUALIFICATION PROCEDURE:
Re-qualification of an Autoclave will be considered qualified for consistent and reliable performance (Qualified)
on successful completion of the following
Vacuum Leak Test
Bowie-Dick Test
Heat Distribution Study
Heat Penetration Study
Biological Challenge Test
Bung Processing (Minimum & Maximum Load)
Machine Parts with Media Fill Parts
Machine Parts with Dress Cycle
Blender Cycle
Media Cycle
To qualify these tests, the equipment should fulfill the acceptance criteria described in the individual test
procedures.
GENERAL CONSIDERATION/ PREREQUISITE:
Approved Standard Operating Procedure of the equipment shall be available
Approved analytical methods for testing the samples collected during the processing
The impact analysis of the equipments shall be recorded in the summary sheet.
All the deficiencies and discrepancies related to the equipment which affect the product quality and corrective
action taken shall be recorded in the appropriate section of the protocol.
The analytical test results and other reports related with the equipment shall be attached with the Performance
Re-qualification of the equipment and finally verified.
CALIBRATION DETAIL OF TEMPERATURE SENSORS:
Equipment
Name
Equipment
ID
Calibration
Record
No.
Due
Date
Date of
Pre
Calibration
performed
Date of
Post
Calibration
performed
Done
By
Checked
By
CALIBRATION OF TEMPERATURE SENSORS:
Pre & Post Calibration of Temperature Sensors.
Pre & post calibration shall be carried out before starting and after completion of heat Distribution cycles as well as
Heat penetration cycles.
PRE & POST CALIBRATION OF TEMPERATURE SENSORS:
PREPRATION OF ICE BATH:
Prepare a container with crushed ice and add enough purified water to ensure a proper Slush Allow the
temperature to stabilize ensure to add sufficient crushed ice to maintain the Equilibrium state of ice and water.
PROCEDURE:
Temperature sensors which are used for qualification study shall be calibrated in ice bath at approximately 0C
and in high temperature reference block at 50, 100, 121, 150C prior to its usage in qualification.
Record the temperature of all the sensors while putting them in ice bath after one minute of temperature
stabilization.
Put the individual sensor to the slot of high temperature reference block which is stabilized at required
temperature.
Record the temperature for five minutes by data logger and attach the report with printouts.
ACCEPTANCE CRITERIA:
The temperature sensors should not vary by 1C in the ice bath, from the mean of temperatures shown by the
calibrated thermometer during the data logging period.
The temperature sensors should not vary by 1C in high temperature reference block, from the mean of
temperatures shown by the calibrated thermometer during the data logging period.
RE-QUALIFICATION CRITERIA:
Replacement of major component/ instrument.
Major modification in the existing equipment/ utility.
During monitoring, if system is found to be malfunctioning.
Shifting of the equipment/ Instrument from one location to another.
SPECIFICATION OF F
0
VALUE AND ACCEPTANCE CRITERIA:
F
0
VALUE: At a Particular time other than 121C, is the time in minutes required to provide the lethality
equivalent to that provided at 121C for a stated time.
F
0
Value of a saturated Steam Sterilization process is the Lethality in terms of equivalent time in Minutes at a
Temperature of 121C delivered by the process to the Product in its final container with reference to
Microorganisms possessing a Z value of 10.
F
0
= t x 10
(T - Tref / Z)
Minutes.
Where,
t = 60 Sec. = 1min.
Tref = 121C
Z = 10C
13.3 ACCEPTANCE CRITERIA:
The calculated minimum F
0
Value (by equation) should be more than or Equal to 18min for all the Thermal
Studies.
i.e., F
0
18min.

CRITICAL VARIABLES TO BE MET:
Sr. No. CRITICAL VARIABLES TO BE MET ACCEPTANCE CRITERIA OBSERVATIONS
1.
Verify all gauges & instruments are
calibrated
Temp. Recorder.
Temp. Controller.
Pressure Gauges.
All calibration data must be
within the acceptable norms of
calibration certificate.

2.
In house calibration
Pre Qualification Studies
Variance: NMT 1
o
C from the
master equipment used.

3. Bowie Dick test kit (No. of cycle: 1)

Run a cycle keeping all other parameters
same, except sterile hold time.
Set the sterile hold time for 8.3 minutes.
Keep the Bowie Dick type test kit as near
as possible to chamber drain in a
perforated support.
Yellow to dark blue with out
any patches.

4. Empty Chamber Heat Distribution Cycle. (No. of cycles: 3)

Set temperature: 121
o
C.
Sterilization time: 30 min.
Sampling Interval: 60Sec.
Through out the Sterilization
phase all temperatures are
with in 3
o
C range.

5. Machine Parts with Dress Cycle (No. of cycles: 3)

Load Size
Machine parts
1. Waste Collector (2 Nos.)
2. Pistons & Nuts with Nylon Tips (8 Nos.),
Guides (16 Nos.)
3. Nozzles gas flushing Units (4 Nos.)
4. Festo Tubes (5 Nos.) & Air Filter (1No.)
5. Vial Separator (2 Nos.)
6. Conveyor Guide (3 Nos.)
7. Collecting Vessel (1 No.)
8. Membrane Filter Holder (1 No.)
9. SS Vessel 4 Lit (1 No.)
10. Teflon Sheets (2 Nos.)
11. Screws and Washers (26 Nos. each)
12. Powder Hopper with Auger & Lid (1
No.)
13. Powder Wheel (1 No.)
14. Bung Hopper (1 No.)
15. Bung Chute (1 No.)
16. Tool Box (1 Set)
Through out the sterilization
phase all temperatures are
with in 3
o
C range.

Initial violet colour of the
Biological indicator should not
change (Yellow) After the
incubation period of 48 hours
at 55 to 60
o
C.



Initial White colour of chemical
indicator should change to
brown or black
Reports from External
Agency.

Biological indicator test
reports attached.


Chemical indicator colour
changes from white
to


Chemical indicator colour
changes from white
to


Chemical indicator colour
changes from white
to


Sr. No. CRITICAL VARIABLES TO BE MET ACCEPTANCE CRITERIA OBSERVATIONS

Dress
17. Sterile Gowns: 18 Nos.
(Six each in 3 Dress Bins).
Set temperature: 121
o
C.
Sterilization time: 30 min.
Sampling Interval: 60Sec.
Biological indicators in all 3 cycles.
Chemical indicators in all 3 cycles.

6. Machine Parts with Media Fill Parts (No. of cycle: 1)

Load Size
Machine parts
1. Waste Collector (2 Nos.)
2. Pistons & Nuts with Nylon Tips (8 Nos.),
Guides (16 Nos.)
3. Nozzles gas flushing Units (4 Nos.)
4. Festo Tubes (5 Nos.) & Air Filter (1No.)
5. Vial Separator (2 Nos.)
6. Conveyor Guide (3 Nos.)
7. Collecting Vessel (1 No.)
8. Membrane Filter Holder (1 No.)
9. SS Vessel 4 Lit (1 No.)
Through out the sterilization
phase all temperatures are
with in 3
o
C range.

Initial violet colour of the
Biological indicator should not
change (Yellow) After the
incubation period of 48 hours
at 55 to 60
o
C.



Initial White colour of chemical
Reports from External
Agency.



Biological indicator test
reports attached.




Chemical indicator colour
10. Teflon Sheets (2 Nos.)
11. Screws and Washers (26 Nos. each)
12. Powder Hopper with Auger & Lid (1
No.)
13. Powder Wheel (1 No.)
14. Bung Hopper (1 No.)
15. Bung Chute (1 No.)
16. Tool Box (1 Set)
17. Media Filling Syringe (1 No.)
18. Festo tubes (Used for Media Fill)
19. Silicon Tubes (2 Nos.)
Set temperature: 121
o
C.
Sterilization time: 30 min.
Sampling Interval: 60Sec.
Biological indicators should be used in all 3
Cycles.
Chemical indicators should be used in all 3
Cycles.
indicator should change to
brown or black
changes from white
to




Chemical indicator colour
changes from white
to




Chemical indicator colour
changes from white
to

7. Bung Cycle (No. of cycle: 1)

Load Size: (Minimum): 5000 Nos.
2 Baskets (Each Baskets should have
Approx. 2500 Bungs)


Set temperature: 121
o
C.
Sterilization time: 30 Minutes.
Sampling Interval: 60Sec.



Biological indicators should be used in
cycle.
Chemical indicators should be used in
cycle.
Through out the sterilization
phase all temperatures are
with in 3
o
C range.


Initial violet colour of the
Biological indicator should not
change (Yellow) After the
incubation period of 48 hours
at 55 to 60
o
C.


Initial White colour of chemical
indicator should change to
brown or black
Reports from External
Agency.
Biological indicator test
reports attached.
Chemical indicator colour
changes from white
to

Chemical indicator colour
changes from white
to

Chemical indicator colour
changes from white
to

8. Bung Cycle (No. of cycles: 3)

Load Size: (Maximum): 66000 Nos.
16 baskets (Each Baskets should have
Approx. 4125 Bungs)



Set temperature: 121
o
C.
Sterilization time: 30 Minutes.
Sampling Interval: 60Sec.


Biological indicators should be used in all 3
cycles.
Chemical indicators should be used in all 3
cycles.
Through out the sterilization
phase all temperatures are
with in 3
o
C range.


Initial violet colour of the
Biological indicator should not
change (Yellow) After the
incubation period of 48 hours
at 55 to 60
o
C.


Initial White colour of chemical
indicator should change to
brown or black
Reports from External
Agency.
Biological indicator test
reports attached.
Chemical indicator colour
changes from white
to
Sign:-
Chemical indicator colour
changes from white
to

Chemical indicator colour
changes from white
to

9. Blender Cycle (No. of cycle: 1)

Load Size
Blender 01 No. ID. No: INJ/674

Container 4 Nos
Sampling Road 01Nos
SS Spoon 01Nos

Set temperature: 121
o
C.
Sterilization time: 30 Minutes.
Sampling Interval: 60Sec

2 Biological indicators per cycle
Chemical indicators per cycle
Through out the sterilization
phase all temperatures are
with in 3
o
C range.
121
o
C 1
o
C +2
o
C
Biological indicator should not
change (Yellow) After the
incubation period of 48 hours
at 55 to 60
o
C.

Initial White colour of chemical
indicator should change to
brown or black
Reports from External
Agency.
Biological indicator test
reports attached.



Chemical indicator colour
changes from white
to

10. Media Cycle (No. of cycle: 1)

Load Size:

Media 30Lit, 7.5Lit 4Nos

Set temperature: 121
o
C.
Sterilization time: 22 Minutes.
Sampling Interval: 60Sec.


02Biological indicators per cycles.
01Chemical indicators per cycles.
Through out the sterilization
phase all location should
showNLT 121
o
C


Biological indicator should not
change (Yellow) after the
incubation period of 48 hours
at 55
o
C to 60
o
C.

Initial White colour of chemical
indicator should change to
brown or black
Reports from External
Agency.
Biological indicator test
reports attached.



Chemical indicator colour
changes from white
to

DOCUMENTS TO BE ATTACHED:
Calibration Reports.
Reports from External Agency on empty chamber heat distribution studies.
Reports from External Agency on heat distribution in machine parts and dress cycle.
Reports from External Agency on heat distribution bung cycle.
Biological Indicator test reports.
SUMMARY:
Validation of Sterilizer with Bung processor was conducted on Date:
Heat distribution and heat penetration studies conducted by the External Agency, expertise in temperature
validation studies in presence of Quality Assurance and Engineering personnel.

From the data generated Heat distribution and Heat penetration is in accordance to the specifications and meet
the requirements of the product, process, safety, regulatory bodies and cGMP obligations.