Process Performance Qualification Protocol for Autoclave

1.0 Objective

The objective of this protocol is to verify the performance qualification attributes i.e. sterilization and to establish sufficient data to assure that the STEAM STERILIZER (Equipment ID No.) supplied by M/s, XYZ is suitable for sterilizing planned load.

• To prepare master chart of sterilization cycle for reference during normal production cycle.
• To demonstrate that Autoclave is leak proof and there is no leakage from the chamber.
• To ensure that system is capable to remove air pocket from Autoclave chamber.
• To ensure that heat distribution throughout the chamber is uniform and the temperature is within the limit of 121 to 123°C.
• To ensure that the heat is sufficiently penetrating into the innermost portions of the load subjected for sterilization to achieve a temperature of 121°C - 123°C during the Sterilization hold period.
• To ensure that the steam Sterilization process, when challenged with Geobacillus Stereothermophilus Biological indicator spore strips having spore population of 10⁶ spores strip, should reduce the bacterial load by more than 6 log reduction.

Related: Low Temperature Sterilization Process (115°C)

2.0 Scope

This Protocol shall be applicable to the Autoclave (Equipment No.: ABCD), situated in the clean room no. A10 in the plant of ……………..Company, located at………………city.

3.0 References

3.1 Internal References

Sr. No.	Reference Detail	Document No.
01	Operation of Kaye Validator
02	Calibration of Measuring & Testing Devices
03	Operation of HTR
04	Operation of Valprobe loggers
05	Handling of Biological Indicator
06	Risk Management
07	Deviation Management

3.2 Others

Sr. No.	Reference Detail	Document No.
01	Scottish Health Technical Memorandum 2010, Part 3 of 6, Validation and verification Sterilization, June 2001	SHTM 2010
02	BRITISH STANDARD BS EN, Sterilization — Steam sterilizers — Large sterilizers (The European Standard EN 285: 1996)	EN 285
03	Industrial Moist Heat Sterilization In Autoclaves PDA Technical Monograph No. 1, 2005 Revision Supplement, Volume 59, Number X	PDA Journal of Pharmaceutical Science and Technology
04	Sterilization of health care products — Moist heat — Part 2: Guidance on the application of ANSI/AAMI/ISO 17665-1	ANSI/AAMI/ISO 17665-1

4.0 Responsibilities

Department	Responsibilities
Quality Assurance	Q.A. Officer / Executive: • To prepare/review standard operating procedure/protocol as per current GMP requirement and organization’s Quality norms. • To ensure & follow correct procedure written in SOP & protocol. • Ensure the sampling instruction written in Protocol is followed to withdraw   sample at specified location in specified time interval and witness of validation activity. • Test the collected product sample tested as per specification. • To report any abnormality to QA Manager / QA   Head and take corrective and preventive action in coordination with respective department head. Q.A. Manager: • Review & Approval of Protocol & Report. • Ensure all the raw data generated were in accordance with protocol & expected results achieved. Head - Q.A./ President-Technology: Review and approval of protocol and validation report.
Technical Department	Executive / Manager Technical: Is responsible for review & scheduling the planned validation activity and to provide technical support for execution of protocol.
Manufacturing	Manufacturing Chemist: To follow instruction written in SOP Production Manager/ Q.A. Manager • Ensure implementation of BMR and protocol. • Ensure the sample were withdrawn & sent to the QC for testing as per specification. • Ensure all the equipment used during Process validation was documented in respective equipment usage log.

5.0 Frequency

5.1 Initial Validation

Three (3) successful runs for all planned loads.
5.1.1 Shifting of the equipment from one location to another location.
5.1.2 If replacement of major components / Instruments.
5.1.3 Change in PLC programme.

5.2 Revalidation

One successful run once in a year for all loads at defined maximum load pattern.

6.0 Rationales for Validation Study

6.1 Rationales for Probes quantity

6.1.1 Empty chamber Heat Distribution

In steam sterilization process steam is not flowing, only condensate formed in the chamber is purged out and the pressure in the chamber is maintained by further inlet of steam. Therefore uniform heat distribution is expected in the steam sterilizer chamber during sterilization hold period and probability of non-uniformity within the chamber is less.

In the Autoclave steam inlet is from the both sides of the chamber through baffles, for uniform distribution of steam in the chamber. Condensate drain point is located at the bottom. Three horizontal planes of two shelves (i.e. top and bottom) are selected for temperature mapping. 4 probes to be placed in four corners of top shelves, 3 probes at corner &1 probe at centre of top selves. 3 probes at corner & 1 probe in drain point of lower shelves (attach layout)

6.1.2 Loaded chamber heat penetration 

For sterilization of Rubber stoppers, Garment & Miscellaneous Load (Silicone tube) the 5 probes are placed inside the load, 1 probe in drain point and 4 probes in the chamber as per respective loading pattern diagram attached as Exhibits to this protocol for monitoring of heat penetration in loaded chamber.

Once the maximum and minimum load are validated for Autoclave at defined validated Load pattern need not to qualify periodically for minimum load. Therefore only maximum loads will be periodically revalidated at defined load pattern considering it as worst case scenario.

6.2 Process Parameter

Key operating parameters as per Operational Qualification identified are as below:

Sr. No.	Parameter Details	Process Parameter
1	Pure Steam Pressure	NLT 2.0 Bar (g)
2	Vacuum	NLT –0.70 Bar (g)
3	Compressed Air	NLT 6.0 Bar (g)

6.3 Functional Risk Assessment

Function wise processes are listed and evaluated for assessment of risk to either product quality and data integrity. It involves mainly following steps.

1. Identifying GMP Risk
2. Identifying Risk Scenarios
3. Assessing the likelihood of An Adverse Event
4. Assessing the severity of impact
5. Detection of adverse impact
6. Overall priority

The above risk were prioritization of risk in High/Medium/Low categories.

6.4 Identifying GMP Risk

System function parameters are evaluated and identified whether they represent a risk when assessed against a series of GMP criteria.
Following types of risks are mainly identified during risk assessment process for qualification of system as below:

• Risks towards non-availability of required documentation
• Risks towards non-availability of required SOPs
• Risks towards non-availability of system Access Control
• Risks towards abnormal user operation performed at the time of system operation
• Risks towards incorrect configuration of system
• Risks towards Improper and/or inadequate training 

6.5 Identifying Risk Scenarios

Having determined that a particular function may have a GMP risk associated with it, the assessment proceeds to identify the various risk scenarios i.e. the events that identify the risks associated with use of the system.
The functions identified are analyzed by considering possible hazards/adverse effects and what controls may be needed to minimize the potential harm.

6.6 Assessing the Likelihood of An Adverse Event

After identifying hazards / adverse events, determine the likelihood (frequency or probability) of it occurring. User considers the likelihood of the adverse event occurring per number of transactions, and assigns category as per estimation of risk.

6.7 Assessing the Severity of Impact

After determining likelihood of adverse event, severity of its impact on process is assessed. These effects take into account impact on regulatory compliance, impact on product quality and impact on data integrity.
The impact of risk occurring may be described as follows:

6.8 Ranking of Adverse Event Severity

Table - I

Value	(S) Severity of Event (Consequence)
3	High(H) : Can cause serious adverse health consequences which can threaten the life of Patient or even death
2	Medium(M) : Temporary or reversible adverse health consequences but the life of the patient is not threatened
1	Low(L) : No effect/Impact for patients

6.9 Detection of Adverse Event

Next step is to identify if the adverse event can be recognized or detected by other means in the system. Adverse event having high probability of detection, may not pose a serious threat because it can be recognized quickly and suitable corrective action taken to mitigate its impact. If an adverse event has a low probability of detection, then the risk condition needs to seriously consider a review of the design or the implementation of alternative procedures to avoid the event.

6.10 Ranking of Adverse Event Detection

Table - II

Value	(D) Level of Detection
1	High (H): The risk can be easily detected through deployed control measure/system and the detection system is automated.
2	Medium (M): The risk can be detected later through deployed control measure/system and the detection is through manual method.
3	Low (L): The risk cannot be detected through deployed control measure/system the detection is possible after longer period/interval.

6.11 Risk Review and Monitor Controls

After controls are implemented, they will be monitored for life cycle of the system. This will be part of performance monitoring of the system. Periodic review after the system is fully operational /validated shall
a. Consider whether previously unrecognized risks are present
b. Determine if previously identified hazards are still present ( and to what level)
c. Ascertain if the estimated risk associated with a hazard is no longer acceptable
d. Evaluate whether all existing controls are still necessary

6.12 Risk Assessment & Control During Execution of study

6.12.1 Nature of Risk & its mitigation action to the risk identified

Sr. No	Description of Risk Identified	Impact		Level of Risk after mitigation plan (Low / Moderate / High)	Expected results	Observation	Tested By (Sign & Date)
Risk of System Access by Unauthorized/untrained personnel
1	Unauthorized person tries to conduct the process.	Process may gets affected.		Low	• Only Qualified persons are authorized to perform the process and conduct the study
2	Untrained operator tries to operate /maintenance of the system.	Process may gets affected.		Low	• Training plan, training record and Operation SOP should be available.
3	Operation SOP does not contain proper information.	Process may gets affected.		Low	• System operation SOP should contain operation information as recommended by QA.
Risk on account of abnormal process condition occurred at the time of system operation
4	Failure of Vacuum leak test	Process gets affected.		Low	• Message appears on PLC “Leak Test Fail”. • SOP contains information to take action accordingly.
5	Failure of Bowie Dick Test	Process gets affected.		Low	• Fail is having indication of light color in center of pattern • SOP contains information to take action accordingly.
6	Failure of Biological Indicator.	Process gets affected.		Low	·
Risk on account of abnormality in connected utilities at the time of system operation
8	If Pure steam pressure Low during running cycle.	Process gets affected.	Low		• Pressure and hence temperature not achieve in the system. • Result in Atypical cycle • SOP contains information to take action accordingly.
9	Low vacuum	Process gets affected.	Low		• Result in Atypical cycle • SOP contains information to take action accordingly.
10	Temperature lower during sterilization running the cycle.	Process gets affected.	Low		• Temperature not achieve in the system. • Result in Atypical cycle • SOP contains information to take action accordingly.
11	Temperature higher during sterilization phase of cycle.	May System loss or damage to components	Low		• Result in Atypical cycle • SOP contains information to take action accordingly.
12	Load is not placed as per defined and validated load pattern	Load may not get sterile	Low		• SOP contains information to take action accordingly. • Authorized and trained person is performing the operation.
Risk occurred in Equipment
13	Door gaskets damaged	Process gets affected.		Low	• Chamber vacuum not maintained • Result in Atypical cycle • SOP contains information to take action accordingly.
14	Malfunctioning of pneumatic valves	Process gets affected.		Low	• Result in Atypical cycle • SOP contains information to take action accordingly.
15	Malfunctioning of Steam trap	Process gets affected.		Low	• Temperature not achieve in the system • Result in Atypical cycle • SOP contains information to take action accordingly.
16	Failure of post calibration of thermocouple.	Could not accessed about product sterilization		Low	• Result of BI will be assessed for probability of survival of microorganisms. • Check physical condition of thermocouples if found damage, white proper justification. • Recalibration to be done.

7.0 Equipments/ Material to be used for Validation Study:

• Calibrated Valprobe loggers/ Thermocouples
• Kaye Valprobe/Kaye Validator
• Calibration Unit
• Biological Indicator
• Media to incubate
• Chart paper (Yokogawa)
• Bowie Dick Pack

7.1 Technical Data

Mfg. By:
Installed on:
Model no.:
Size:
Working Pressure:
Volume:
Door:
Hydraulic test Pressure:
Construction:

The Autoclave consists of the following features.

• The Autoclave Chamber is made up of Stainless Steel sheet, which is welded with a U-Profile Stainless Steel Jacket. The Autoclave Chamber is provided with two sliding doors, which are also made up of Stainless Steel reinforced with mild steel support structure. The door is operated with the help of pneumatic cylinder, when the door reaches the end position gaskets are pushed out automatically with help of compressed air for sealing similarly to open the door gasket is retracted by vacuum. When the gasket is retracted the door slides automatically. The Door sealing is done with the help of tubular, silicone rubber gasket. To ensure proper sealing the gaskets are activated with compressed air and retracted with the help of vacuum.
• Door interlocks are provided to prevent simultaneous opening of both the doors of critical area side (sterile area side) and controlled area side (non- sterile area side) and process lock to prevent opening of the door during the operation.
• The Autoclave chamber is insulated with resin-bonded glass wool, which helps in reducing the heat loss to the environment and ensuring uniform distribution of temperature inside the chamber. This insulation is covered with stainless steel cover plate.
• A Stainless steel pipe stand support is provided for the equipment thus requiring no special foundation. For ensuring leak tight partition between the Controlled area (Non-Sterile area side) and Clean area (Sterile area side), a Stainless steel flush paneling is provided on the partitioning wall and the outer cover of Autoclave. All Joints, Crevices are filled with Silicon sealant to prevent any leakage.

The Autoclave is provided with the following systems and accessories for proper functioning.

• Water-ring type vacuum pump with suitable electric motor.
• Vacuum break filter on the clean area side. One pair of removable Stainless Steel railings inside the sterilization chamber for smooth and easy operation of the load article.
• Loading tray constructed of Stainless steel SS316.
• Chamber Compound gauge on sterile and Controlled area side, Jacket pressure gauge of the Controlled area side, Gasket compound gauge for both the doors on the Controlled area side and critical area side, Safety valve for jacket and chamber, Steam trap with strainer and NRV for chamber.
• Pure steam is supplied through SS-316 piping from generation center installed at WFI generation room.
• Control panel consisting of PLC, Digital Temperature input to the microprocessor from a 5 No’s of Temperature transmitters, one Pressure transmitter, Pressure and vacuum switches for giving digital pressure/vacuum signals to the microprocessor, Manual backup system for operation in case of microprocessor failure.
• Stainless steel Internal Piping, Ball valves with rotary actuators for all the process lines with solenoid valves to regulate air supply to the pneumatic rotary actuators.

7.2 Monitoring and Controlling of the sterilization cycle

7.2.1 Two PT-100 probes, inserted in Chamber to monitor temperature on yokogawa recorder.
7.2.2 One similar PT-100 probe is left in the drain by which cycle is controlled by PLC. Temperature of drain shall be monitored on PLC and at yokogawa recorder.

7.3 Study Design

S. No.	Study	Name of the Recipe	No. of trials to be taken
1	Vacuum leak test	Leak Test	One
2	Bowie – Dick Cycle	Bowie-Dick	One
3	Empty chamber Heat Distribution	HPHV RSD	One
4	Loaded chamber with Rubber Stoppers	HPHV RSD	One
5	Loaded chamber with garment and hand gloves	HPHV RSD	One
6	Loaded chamber with Miscellaneous items (Silicone tube)	HPHV RSD	One

Note: - New load may be introduce in loading pattern in between two scheduled re- qualification study after three successful validation run and report of same shall be attached as addendum to last validation report.*

Reference cycle detail (Initial validation with Protocol No)

Sr. No.	Protocol No.	Protocol No.
1	Heat Penetration study in Rubber closure RFS (Ready For Sterilization) bag (Applicable for 20 mm rubber stopper)
2	Heat Penetration study for Garment Sterilization
3	Heat Penetration study for Miscellaneous Load

7.4 Sterilization Cycle Parameters

Sr. No	Name of Parameter
		Leak test
1.	Programme Name	Vacuum Leak Test
2.	Select Cycle (Recipe No)	4
3.	Leak Vacuum on	-0.7 kg/cm2
4.	Vacuum Hold	15 min

7.5 Recipe For Bowie Dick Cycle

Sr. No	Name of Parameter	Bowie – Dick Test
1.	Select Cycle (Recipe No)	5
2.	Initial vacuum time	2 min
3.	Pulsation	3
4.	Preheating Temperature	95ºC
5.	Preheating Time	1 min
6.	Sterile Temperature	121.0 ºC
7.	Sterile Time	10 min
8.	Sterile Print Time	1 min
9.	Dry Vacuum Time	0 min
10.	Dry Vacuum Value	-0.700 Kg/cm2
11.	Door Open Temperature	90 deg

7.6 Recipe For HPHV RSD cycle

Sr. No	Name of Parameter	Value
1.	Select Cycle (Recipe No)	7
2.	Initial Vacuum Pulse	3
3.	Initial Vacuum Value	500 mm of Hg
4.	Initial vacuum time	2 min
5.	Preheating Temperature	95 ºC
6.	Preheating Time	1 min
7.	Sterile Temperature	121.0 ºC
8.	Sterile Time	15 min
9.	Sterile Hold Print Time	60 sec
10.	Dry Vacuum Pulse	3
11.	Dry Vacuum Value	600 mm of Hg
12.	Dry Vacuum Time	20 min
13.	JKT STM PRE REL	2 min
14.	Cool Vacuum Value	600 mm of Hg
15.	Cool Vacuum Time	5 min
16.	Jacket Air vent ON	5 sec
17.	Jacket Air vent OFF	12 sec
18.	Jacket Air Pressure Rel	1 min
19.	Door Open Temperature	90 deg

8.0 Procedure

8.1 Vacuum leak test Procedure

8.1.1 Put the flexible probes inside the chamber through the validation port provided for the validation cycles prior to start of Vacuum leak test.
8.1.2 Ensure that Autoclave is empty and chamber is at ambient temperature.
8.1.3 Ensure compressed air is ‘ON’ at required pressure of 6 bar.
8.1.4 Ensure sufficient water supply is available for seal of vacuum pump
8.1.5 Ensure gasket lubrication is proper
8.1.6 Switch ‘ON Main’ switch provided on panel board.
8.1.7 Close loading side door by pressing “Door 1 close” Push button provided on main panel board.
8.1.8 Ensure chart recorder is ‘ON’ & graph is properly loaded.
8.1.9 Check the programme from control panel.
8.1.10 Enter the Operator level password & enter the programme as per requirement.
8.1.11 Select programme by pressing ‘Enter’ Key F1 once.
8.1.12 All parameter will start on displaying on screen one by one.
8.1.13 Enter F1 button and select programme, the display will show YES-1 NO-2.
8.1.14 Checks the online printer attached to PLC & then select 1 and press Enter.
8.1.15 Cycle will start in automatic mode as per set parameter as PLC will show “Leak Test Vac ON” and vacuum will be created in chamber.
8.1.16 After completion of hold period, vacuum release valve will open and message will display “Leak Test Vac Release” and continue till the chamber pressure comes to atmospheric pressure.
8.1.17 At the end of the cycle, if the leak rate is more than 1 mm of Hg/ min during hold period, then message will display “Leak Test Fail” & if it is less than 1 mm of Hg/min during hold period, then message will display “Leak Test Pass”.
8.1.18 If cycle fails, initiate work request form for rectification of problem.
8.1.19 After rectification of problem fresh cycle to be run. Upon completion of cycle screen will display “Cycle Over”.
8.1.20 Continue to run for next cycle of Bowie-Dick test.

8.2 Bowie Dick test Procedure

8.2.1 Load the tray in the autoclave chamber.
8.2.2 Place the Bowie-Dick test pack on the bottom shelf of the sterilizer just above the drain point (Nearly 100 mm over the drain).
8.2.3 Selected the recipe Bowie-Dick test cycle from MMI/HMI display as per point no 8.1.10 to 8.1.12. Ensure that sterilization hold time is 10 minutes, at 121°C. (Manufacturer’s recommendation).
8.2.4 Start the cycle by pressing Enter key.
8.2.5 After the cycle is over, open the door from control area side & take the sterilized test pack from the Autoclave and check the indicator paper/strip for uniform color change.
8.2.6 Observation should be recorded in test data sheet no 05 (Exhibit II).
8.2.7 A Fail is indicated by light color in the center of the pattern than around the edges when compared with control.
8.2.8 Failure of the pattern to change completely black or dark brown at 121 °C when compared with control.

8.3 Procedure For Empty Chamber Heat Distribution Study

8.3.1 Connect the calibrated thermocouples with SIM to the Kaye Validator and distribute these thermocouple through out chamber as per Exhibit-III.
8.3.2 Operate Kaye Validator as per SOP or Val probe logger as per SOP.
8.3.3 Run HPHV RSD cycle as per SOP with recipe defined in point 7.6.
8.3.4 Select the cycle from the MMI/ HMI panel as per point no.8.1.10 to 8.1.12 & start HPHV RSD cycle as well as start Kaye Validator at same time. Scan data at every 10 sec interval in Validator.
8.3.5 After completion of the cycle take the printout of report generated through Kaye Validator. Evaluate the report as per acceptance criteria.
8.3.6 Attach the report generated through Kaye Validator and PLC print out & graph from chart less recorder to the report.

8.4 HEAT PENETRATION STUDIES

8.4.1 Procedure For Loaded chamber heat penetration study:
8.4.1.1 Load the articles into Double door Autoclave as per their respective loading diagram attached as exhibit to this protocol.
8.4.1.2 Place heat penetration probes inside the loaded articles and select the positions where heat penetration is difficult.
8.4.1.3 Connect the Thermocouple with SIM to Kaye Validator as per SOP or Val probe logger as per SOP, which senses the temperature.
8.4.1.4 Place biological indicator folded with aluminum foil strips of Geobacillus Stereothermophilus containing 106 spores along with flexible probes.
8.4.1.5 Select the HPHV RSD cycle as per point no. 8.1.10 to 8.1.12.
8.4.1.6 Start the cycle by pressing Enter key from the MMI / HMI panel as well as start Kaye Validator at same time. Scan data at every 10 sec interval in Validator.
8.4.1.7 After completion of the cycle take the printout of report generated through Kaye Validator. Evaluate the report as per acceptance criteria
8.4.1.8 Carry out total three replicate runs with loaded chamber for newly introduced load (first time study).
8.4.1.9 Take out all exposed biological indicator strips & send the biological indicator to microbiological lab for testing as per SOP of handling of biological indicators and recording the results in observation sheet.
8.4.2 Heat Penetration Study: Maximum Load of 20mm Rubber Stopper in RFS Bag (Ready For Sterilization)
8.4.2.1 Load the 20 mm rubber stopper [(1) 5000 Nos. x 5 bags = 25,000 nos. OR (2) 2500 nos. x 10 bags = 25,000 nos.] packed in Rexam pack ready for sterilization in the chamber of autoclave in two trays as per defined loading diagram attached with this protocol.
8.4.3 Heat Penetration Study: Minimum Load of 20mm Rubber Stopper in RFS Bag
8.4.3.1 Load the 20 mm rubber stopper (2500 nos. x 1 bags = 2,500 nos.) packed in Rexam pack ready for sterilization in the chamber of autoclave in two trays as per defined loading diagram attached with this protocol.
8.4.4 Heat Penetration Study: Garment Sterilization with Maximum Load
8.4.4.1 Load the 32 numbers of garment packed in baby bag & 32 goggle for sterilization in the chamber of autoclave in two trays as per defined loading diagram attached (Exhibit VIII) with this protocol.

8.5 Heat Penetration Study: Garment Sterilization with Minimum Load

8.5.1 Load the 5 numbers of garment packed in baby bag & 5 goggle for sterilization in the chamber of autoclave in two trays as per defined loading diagram attached (Exhibit IX) with this protocol.

8.6 Heat Penetration Study: Miscellaneous Load (Silicone Tube)

8.6.1 Ensure cleaning of Silicone tube before packing in Rexam bag.
8.6.2 Keep the silicone pipe (Approx 2 meter long) in Rexam bag by folding in circular way, keeping both end open.
8.6.3 Insert the biological Indicator folded with aluminum foil along with Thermocouple in both end of silicone tube. Insert the probe as long as it goes inside the tube. Care should be taken when insertion that after sterilization the biological indicator should be removed easily.
8.6.4 Load the 4 number of silicone tube packed in Rexam pack ready for sterilization in the chamber of autoclave in two trays as per defined loading diagram attached with this protocol.

8.7 Load Configuration

Sr. No	Load No	Name of the Load	Load Pattern	Load Type	No of Articles
01	NA	Leak Test	NA	NA	NA
02	NA	Bowie Dick Test	Bowie Dick	NA	1 No
03	1	Rubber Stopper	20 mm Rubber Stopper (5000 nos x 05 bags) or (2500 nos x 10 bags)	Maximum	25,000 Nos
04	3	Rubber Stopper	20 mm Rubber Stopper (2500 nos x 1 bag)	Minimum	2,500 Nos
05	10	Garment Load	Garment Bags	Maximum	32
			Goggle		32
06	11	Garment Load	Garment Bags	Minimum	5
			Goggle		5
07	12	Miscellaneous Load	Silicon tube	NA	4

Note: During validation study, if there is any change required in the type and/ number of articles to be loaded, loading diagram will be changed. As per the validation result the diagrams will be established for routine cycles and accordingly SOP will be established. In this case protocol will not be revised, as validation method will remain same.

8.8 Attachments to Report

8.8.1 Attachment 01: Common for all tests

• Calibration Report of Thermocouples
• Calibration Verification Report of Thermocouples

8.8.2 Attachment 02: For Leak Test Cycle

• Yokogawa Graph
• PLC print

8.8.3 Attachment 03: For Bowie-Dick Test

• Test Indicator Paper
• Yokogawa Graph
• PLC print

8.8.4 Attachment 04: For Heat Distribution Study

• Standard Format: VAL/STR/1.4
• Yokogawa Graph
• PLC print
• Thermocouples set up Report
• Thermocouples Qualification Report

8.8.5 Attachment 05: For Heat Penetration Study for 20mm Rubber Stopper Maximum Load

• Standard Format: for logging data
• Yokogawa Graph
• PLC print
• Thermocouples set up Report
• Thermocouples Qualification Report

All above documents are required for below attachments as per respective studies:

• Attachment 06: For Heat Penetration Study for 20mm Rubber Stopper Minimum Load
• Attachment 07: For Heat Penetration Study for Garment Sterilization Maximum Load
• Attachment 8: For Heat Penetration Study for Garment Sterilization Minimum Load
• Attachment 9: For Heat Penetration Study for Miscellaneous Load

9.0 Sampling Procedure

• Scan data at every 10 sec interval in Validator/Valprobe.
• Minimum 10 nos. of temperature sensors placed in the chamber geometrically for temperature monitoring study.
• Minimum 10 nos. of Biological Indicator placed in the chamber next to the temperature sensors for temperature monitoring study.

10.0 Acceptance Criteria

10.1 Vacuum Leak Test

10.1.1 The vacuum drop should not be more than 1mm Hg/ 1 min of vacuum hold.

10.2 Bowie-Dick Test

10.2.1 The indicator paper / sheet should be uniform change in color (Brown) over the entire pattern of indicator sheet when compared with control.

10.3 Heat Distribution Study

10.3.1 Temperature distribution within the chamber must be between 121°C to 123°C at all location during the sterilization period (dwell time).
10.3.2 There should not be any slowest heating point (Cold Spot) in the Autoclave chamber.
10.3.3 The equilibrium time should not be more than 30 sec.

10.4 Heat Penetration Study

10.4.1 Temperature distribution within the chamber must be between 121°C to 123°C at all location during the sterilization period (dwell time).
10.4.2 Sterilization temperature should be maintained for NLT 15 min for minimum 10 thermocouple during hold period.
10.4.3 6-log reduction should be observed in any exposed biological indicators on complete incubation period.
10.4.4 The equilibrium time should not be more than 30 sec.

10.5 Action to be taken in case of failure

10.5.1 If the vacuum leak test fail, this may be due to insertion of thermocouple, initiate the work request for the maintenance of autoclave leakage.
10.5.2 If the Bowie-Dick test fails, this may be due to gasket leakage of leakage due to insertion of thermocouple. Initiate the work request for the maintenance leakage of autoclave.
10.5.3 If the Heat penetration or Heat Distribution cycle fails in any respect, raised the deviation as per SOP. Discard the cycle, investigate the cause of failure. Make the required corrections / modifications by initiate the work request for the maintenance of equipment prior to carrying out three additional sterilization cycles.
10.5.4 During the revalidation if any load configuration fails in any respect a deviation should log & load configuration should be invalidated. After through investigation, three runs of same load / modified load configuration to be validated with proper justification.
List of Standard Operating Procedure involved during validation study:

Sr. No.	SOP title	SOP No.	Issue No. / Revision No.
1.	Sterilization Procedure		Issue No. : Rev. No.  :
2.	Handling of biological Indicator		Issue No. : Rev. No.  :
3.	Operation of Kaye Validator 2000		Issue No. : Rev. No.  :
4.	Calibration of Measuring & testing devices		Issue No. : Rev. No.  :
5.	Operation of Val probe logger		Issue No. : Rev. No.  :
6.	Operation of HTR-400		Issue No. : Rev. No.  :
7.	Sterilization Procedure		Issue No. : Rev. No.  :

Details of Equipment used for analysis

Sr. No.	Name of Equipment	Equipment No. / ID No.	Last Calibration / Validation done on	Next Calibration / Validation due on	Remarks
1.	Kaye Validator 2000
2.	Val Probe logger

11.0 New risk observed if any During the process

Pl. mention as it is report part

12.0 Discussion

12.1 Procedure: To mention, how conducted the study. Pl. mention as it is report part
12.2 Acceptance Criteria: Pl. mention as it is report part
12.3 Change Control Note: Study is conducted under any change control system , if yes, pl. mention.
12.4 Deviation: Pl. mention for deviation, if any.
12.5 Out of Specification: Pl. mention for OOS, if any.

12.6 Within Date:

12.6.1 Last Validation performed on: _________________
12.6.2 Current Validation performed on: _________________
Remarks (if any):
12.7 Training: Pl. mention about training details given for protocol execution to team members.

13.0 Modifications Suggested

After execution of protocol, if any modification is suggested to get the desired results.

14.0 Conclusion

Conclude the report for acceptance or non acceptance of equipment as per protocol acceptance criteria.

15.0 Records

15.1 Standard Formats

Sr. No.	Format Detail	Document No.
15.1.1	Records of All The Cycles/ Runs With Relevant Calculations
15.1.2	Bacterial Challenge Test Report For Heat Penetration Study

15.2 Exhibits

Sr. No.	Exhibit Detail	Exhibit No.
15.2.1	Front View Of Sterilizer
15.2.2	Bowie-Dick Test Load
15.2.3	Temperature sensor distribution in Empty Chamber Heat Distribution Cycle
15.2.4	Temperature sensor & BI distribution For 20mm Rubber Stopper Maximum Load
15.2.5	Temperature sensor & BI distribution For 20mm Rubber Stopper Minimum Load
15.2.6	Temperature sensor & BI distribution For 32mm Rubber Stopper Maximum Load
15.2.7	Temperature sensor & BI distribution For 32mm Rubber Stopper Minimum Load
15.2.8	Temperature sensor & BI distribution For Garment Maximum Load
15.2.9	Temperature sensor & BI distribution For Garment Minimum Load
15.2.10	Temperature sensor & BI distribution in Miscellaneous Load	Exhibit-X

Also see: Autoclave Validation in Pharmaceuticals