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Performance Qualification Protocol of TUNNEL STERILIZER

DEPARTMENT PRODUCTION
NAME OF THE SUPPLIER
MODEL
SERIAL NUMBER
LOCATION VIAL WASHING ROOM
EFFECTIVE DATE

 

CHANGE HISTORY
Date Supersede CCR No. Changes Made Revision No.
Nil NA Original Issue 00

 

TABLE OF CONTENTS
S. No. Description Page No.
1.0 Protocol Approval
2.0 Objective
3.0 Scope
4.0 System Description
5.0 Responsibilities And Identification Of Execution Team
6.0 Test Procedures
7.0 Recording Of Observations
8.0 Discrepancy And Corrective Action Report
9.0 Compilation, Review And Summary Report
10.0 Appendix
11.0 Requalification Criteria
  • PROTOCOL APPROVAL

Signing of this approval page of protocol indicates agreement with the qualification approach described in this document. If any modification in the equipment qualification approach becomes necessary, a revision through change control shall be prepared, checked and approved. This protocol cannot be executed until approved by following personnel. 

Department Name Designation Signature /Date
Prepared by
Quality Assurance
Reviewed by
Production
Quality Control
Approved by
Quality Assurance
Authorised by
HOD – QA

 

  • OBJECTIVE
    • The objective of this protocol is to provide high degree of assurance in the sterilization and Depyrogenation process, using Tunnel sterilizer supplied by _______________________. The sterilizer is used to sterilize/depyrogenate glass vials (both moulded and tubular) ranging from 5ml vial to 20ml vials.
    • Consecutive 3 runs for Heat penetration studies shall be carried out on each type of vial. These heat penetration studies shall include Bacterial Endotoxin challenge study using Bacterial Endotoxin spiked vials (containing 104 EU/vial). The placement of the vials with Endotoxin indicator vials in the vial load under test shall be in such a way that simulation of the worst-case consideration and normal load pattern can be established at a time.
    • Estimation of the FH value achieved during the sterilization hold period at each temperature-mapping probe. Biological indicators are not used as the intended cycle is for depyrogenation at a very high temperature and survival of the microorganisms is not possible after the depyrogenation cycle.
  • SCOPE
    • This protocol is applicable to Tunnel sterilizer installed in the vial washing room of Production (sterile) area.
    • To be performed after the completion and authorization of Operational Qualification.
    • To be performed at the time of relocation or Requalification.
  • EQUIPMENT DESCRIPTION
  • Sterilizing and depyrogenating tunnel enables the integration of the process of sterilization and depyrogenation of washed empty vials for sterile, continuous automatic filling in which all steps are performed under Grade A conditions.
  • The Tunnel is comprised of four zones, i.e. Drying zone, sterilizing zone, cooling zone and Stabilizing zone.
  • Drying, cooling and stabilizing zones are essentially once-pass-through Laminar flow units comprising as standard features, Pre-filters, motor blower assembly and HEPA filters.
  • Extract and exhaust blowers are provided under the conveyor connected to ambient using a common duct.
  • Vials are transferred by a stainless steel Wire Loop conveyor through drying, sterilizing, cooling and stabilizing zones under non-turbulent class ISO 5 fresh air.
  • Container-controlled partition wall adjustment panels are provided on entry and exit of sterilizing zone.
  •  In the In feed area air taken from the room is prefiltered and aspirated by a blower through HEPA filter. Laminar flow of air is sent vertically down on to the containers.
  • Containers passed nearer to the sterilizing zone they are preheated up to 90-110°C by the hot air bleeding out of the sterilizing zone .the air along with water vapour is picked up underneath of the conveyor belt by the extract blower and ejected to out side.
  • Dried containers are moved over to the sterilizing zone. Sterilizing zone mainly comprises of resistance heating elements, HEPA filters and temperature regulating devices.
  • Hot filtered air is re-circulated in the sterilization zone, which sweeps the containers from top to bottom.
  • Differences exist in the hot air temperature and conveyor speed settings depending on the total mass of the glass required to be sterilized and cooled.
  • Equal pressure drop across the filter enables in creating an extremely even distribution of air circulation.
  • This uniformly distributed hot air sterilizes and depyrogenates the containers. In the cooling and stabilization zones, containers are subjected to Laminar flow of HEPA filtered air taken from the room.
  • The airflow is profiled in such a way that the glass temperature as it excited the sterilizing zone would be transitioned at a nearly linear rate.
  • Stabilizing zone is used as a transition area separating the critical filling area and washing area.
  • Conveyor system is controlled by the container accumulation at the infeed and the set temperature for conveyor start.
  • SS wire Loop conveyor driven by a gear motor with AC frequency controlled drive and tension roller for maintaining the correct tension of the conveyor belt.
  • Control and recording systems for the parameters governing the sterilizing process such as air temperature, belt speed, holding time are standard features.
  • Sterilizing and depyrogenating tunnel is equipped and controlled with an Allen Bradley PLC and MMI (Man Machine Interface) which in conjunction with RTD’s, Infeed / Outfeed Proximity switches, Dot Matrix printers and frequency converter (Variable speed drive) make the operations fully automatic.
  •  Mode of operation enables to opt a) Auto mode b) Maintenance mode.
  • In the Auto mode the operation of the tunnel is fully automatic. Tunnel starts and stops at Predetermined time. All blowers are running continuously except sterilizing zone heaters, which cuts off when temperature reaches set temperature.
  • In the Maintenance mode all operations are being done manually.
  • RESPONSIBILITIES AND IDENTIFICATION OF EXECUTION TEAM
    • Responsibilities: The group comprising of representatives from each of the following departments and they shall be responsible for the overall compliance with this protocol. 
Department Responsibility
Production Execute the validation activity & provide necessary support
Engineering & Utility Participate & provide necessary support for the validation activity
Quality Control Testing of samples as per the test procedures
Quality Assurance Monitoring, sampling & reviewing the validation activities.
  • Identification of Executors: The personnel involved in the execution of this protocol shall be recorded Name, Designation, Signature and Date.
  • TEST PROCEDURES

Following test procedures are followed to qualify the equipment for its performance.

  • Measurement of Airflow Velocity
    • Tools required: Calibrated Anemometer.
    • Test procedure:
      • Ensure that the equipment is ON except heating prior to the start of the observations before 30 minutes.
      • Calculate the number of location by using the following formula. Distribute all the locations equally for each filter outlet.

Number of locations = square root of 10 times of area in square metres but no less than 4, we have considered 4 sampling points for each filters.

  • The airflow velocity should be measured at approximately 150 mm to 300 mm from the filter face by using calibrated Anemometer. The measuring time at each location should be also sufficient to ensure a repeatable reading.
  • Record minimum 3 readings in each location and calculate the average measured velocity. Record the calibration details of the instrument used for measurement and attach the calibration certificate.
  • Acceptance criteria:
    • For Drying zone: Airflow velocity across the HEPA filter should be within the range of 98 ± 20 feet / minute [0.5± 0.1 m/sec]
    • For Sterilising zone/Heat zone: Airflow velocity across the HEPA filter should be within the range of 157 ± 20 feet / minute [0.8± 0.1 m/sec].
    • For Cool zone: Airflow velocity across the HEPA filter should be within the range of 118 ± 20 feet / minute [0.6± 0.1 m/sec].
  • Differential Pressure Test
    • Tools required: Calibrated Magnehelic Gauge / Photohelic gauges.
    • Test procedure: Observe the differential pressure across the zones through Magnehelic Gauge / Photohelic gauge and note down the reading before operation and during operation. Repeat the procedure for two more times.
    • Acceptance criteria:
      • Magnehelic gauge should show the reading at ‘0’ before operation.
      • During operation,
    • For Drying Zone: equal to or above2 MMWG.
    • For Sterilising zone: equal to or above 25 MMWG
    • For Cool zone: equal to or above 25 MMWG
      • Installed Filter System Integrity and Leakage Test (DOP/PAO Test)
        • Tools required: Aerosol generator, Aerosol Photometer (Calibrated) and Dioctyl pthalate or PAO Liquid (Poly Alpha Olefin).
        • Test procedure:
          • Operate the Tunnel steriliser as per the SOP.
          • Position the Aerosol generator and introduce Aerosol into the upstream air, ahead of the HEPA filters, at the concentration of 80-100 mg per litre of air at the filter’s designed airflow rating and set the instrument at 100% concentration.
          • Scan the downstream side of the filter with probe at scanning rate not to exceed 15cm2/Second. The probe should be held at a maximum distance of 5cm from the down stream filter face. Scanning should be performed over the entire downstream face of each filter, the perimeter of each filter, the seal between the filter frame and the grid structure, including its joints. Measurements of the aerosol upstream of the filters should be repeated at a reasonable time intervals between and after scanning for leaks, to confirm the stability of the challenge aerosol concentration.
          • Record the observations. Record the details of the instruments used including its calibration status, filter identification number and upstream and downstream concentration of Dioctyl pthalate/ PAO. Attach the calibration certificate of the photometer.
        • Acceptance criteria: Designated leaks are deemed to have occurred where a reading greater than 10−4 (0.01%) of the upstream challenge aerosol concentration. Alternative acceptance criteria may be agreed based on the filter efficiency.
      • Airflow Pattern Test
        • Tools required: Dry Ice.
        • Test procedure: Take Dry Ice and check for airflow direction in the zones. Observe the direction of the flow and the same shall be recorded.
        • Acceptance criteria:
          • The stream of air should be unidirectional and non-turbulent up to working zone.
          • Drying zone: airflow towards the vial washing side.
          • Sterilising zone: airflow towards the drying zone.
          • Cool zone: airflow towards Cooling zone and airflow from Vial filling side.
        • Non Viable Particle Count Test
          • Tools required: Calibrated Air borne particulate counter.
          • Test procedure:
            • Calculation of Measuring Points (Sampling Locations)
              • Calculate the number of location by using the following formula. Distribute all the locations equally in the clean zone.

                                         Number of locations = Square route of the area in meters

  • Sampling:
    • Tunnel steriliser blowers shall be in continuous operation for at least 30 minutes prior to performing these tests. AHU of the particular room should also be in operation.
    • Sampling shall be done at rest condition at pre-determined locations as per annexure-1.
    • The non-viable particulate counts shall be done before and after the de-pyrogenation cycle. If the trails are conducted continuously on the same day, then the particulate count are taken before starting the first trial and after the completion of last trial.
    • Set the particle counter in ISO mode. Position the sampling probe towards airflow at work level and Sample at least 3 cycles for each location. Each cycle should be Approx. 28 litres/minute.
    • Paste the printouts generated by the particle counter on A4 page and shall be photocopied.
    • Verify the printouts for all calculations and acceptance criteria as mentioned below. Attach the photocopies of the same to the annexure –1. Attach the calibration certificate of Particle Counter.
  • Acceptance criteria:
    • All the readings of test should pass the below mentioned criteria.
    • Maximum permitted number of particles/m3 equal to or above 0.5μm is 3500 and 5μm is 20.
  • Heat Distribution Study For Empty Chamber
    • Tools required: Calibrated Data logger
    • Test procedure:
      • Operate the tunnel steriliser as per the SOP.
      • 10 numbers of temperature sensors shall be passed randomly along with the conveyor from the drying zone. Ensure that the entire conveyor area to be covered during mapping. The placement temperature sensors in empty chamber. The natural progression of sensor on the conveyor that occurs during actual running of the tunnel shall be simulated.
      • Record the set parameters for the sterilization/Depyrogenation cycle to be operated during the test.
      • Record the position of temperature sensors in representative schematic form.
      • Connect the probes to suitable data logger, which can scan and print the actual temperatures with respect to time.
      • Start the data logger to record the actual temperatures within the sterilization zone with respect to time.
      • When the sterilization / Depyrogenation cycle completes:
    • When all the temperature sensors cross the cooling zone, Stop the conveyor belt of sterilizing tunnel, switch off the data logger and pull out the probes.
    • Collect print out from the printer of the tunnel control panel and attach to the annexure-1 with attachment No.
    • Download the data from the data logger into the computer for data analysis and printing. Review and attach the printouts with attachment No.
      • Repeat two more consecutive runs to demonstrate sterilization cycle reproducibility.
    • Acceptance criteria
      • There should be uniform temperature distribution across the conveyor belt during the hold period.
      • The temperature at each temperature sensor should be 325°C ± 250C for NLT 1 Minute during the depyrogenation cycle.

Note: The temperature sensors shall be placed in straight line across the belt, tied to the SS wire, during heat distribution studies.

  • Heat Penetration Study For Load Chamber
    • Tools required: Calibrated Data logger
    • Test procedure:
      • Operate the tunnel steriliser as per the SOP.
      • Empty glass vials (as required) shall be fed to the tunnel sterilizer from the vial washing machine.
      • 10 numbers of Vials spiked with known amount of Endotoxin (Endotoxin indicator vials) as per SOP and 10 numbers of empty vials with temperature sensors shall be placed in the load as depicted below, to simulate normal sterilization and simulated worst condition in a single load. Both the vials with endotoxin and temperature sensor must be placed side by side.
      • The placement Endotoxin indicator vials and temperature sensors in empty vials . The vials shall not be tied with wire to keep together. The natural progression of vial on the conveyor that occurs during actual running of the tunnel shall be simulated.
      • Record the set parameters for the sterilization cycle to be operated during the test.
      • Record the position of the vials with temperature sensors and vials with Endotoxin in representative schematic form.
      • Connect the probes to suitable data logger, which can scan and print the actual temperatures with respect to time.
      • Start the data logger to record the actual temperatures within the sterilization zone with respect to time.
      • During the validation run inspect the vials in the tunnel sterilizer for the following parameters:
    • Look through the glass window of the tunnel to following shall be verified
    • Position of the vials on the conveyor.
    • Movement of the vial on the conveyor belt.
    • To inspect the incoming vials to the in-feed turntable of the powder filling machine.
    • Breakage / Cracks in vials on sterilization.
    • Presence of any spot on sterilized vial surface.
      • When the sterilization cycle completes:
    • When all the vials, attached with temperature indicating probe and endotoxin indicator vials, and vials crosses the cooling zone, Stop the conveyor belt of sterilizing tunnel, switch off the data logger and pull out the probes.
    • Collect print out from the printer of the tunnel control panel, review and attach to annexure-1 with attachment No.
    • Download the data from the data logger into the computer for data analysis and printing. Review and attach the printouts to annexure-1 with attachment No.
    • Wrap the exposed endotoxin indicator vials with sterile aluminium foil in filling room and identify suitably. Send the exposed vials to Microbiology lab to estimate the quantity of the residual endotoxin as per the SOP. Record the results in the annexure-2.
      • Repeat two more consecutive runs to demonstrate sterilization cycle reproducibility.
    • Acceptance criteria
      • There should be uniform temperature distribution across the conveyor belt during the hold period.
      • The temperature at each temperature sensor should be within ± 250C during the depyrogenation cycle.
      • FH value should be more than the value calculated in the FH value calculation sheet proving the sterility assurance by 12-log reduction. (Using assumed D-value and worst case heat resistant spore population)
      • The temperature distribution is uniform throughout the sterilization period i.e. 325°C ± 250C for NLT 1.0 min.
      • Endotoxin spiked vials exposed to the depyrogenation cycle should show more than 3-log reduction in the endotoxin content when tested.
      • Conveyor should be packed with vials.
      • Vial should move on the conveyor belt in erect condition. No vial should be tilted.
      • No breakage of vial should be observed.
      • No charring spot should be observed on the surface of the vials.
    • RECORDING OF OBSERVATIONS

Record the observations of after execution of each test procedures, in the annexure –1 (Recording of Observations For Performance Qualification).

  • DISCREPANCY AND CORRECTIVE ACTION REPORT

Document any discrepancies observed during the Performance qualification of the equipment in annexure -1. Include the corrective actions of the same. When all the discrepancies are satisfactorily resolved or an approved action plan is developed which ensures that the discrepancy will be resolved.

  • COMPILATION, REVIEW AND SUMMARY REPORT

Compile and review that all test functions have been completed, reconciled and attached to this protocol. Verify that the approvals for deviations have been taken and are resolved appropriately to the satisfaction.

Performance Qualification shall be considered acceptable when all the conditions specified in the test procedures have been met.

Prepare the summary report in the annexure –2 (Performance Qualification Report) and submit this for review, approval and authorisation to Validation Core Team.

  • APPENDIX
    • Abbreviations and definitions
Abbreviation Definitions
PQP Performance Qualification Protocol
SOP Standard Operating Procedure
QA Quality Assurance
QC Quality Control
NA Not Applicable
No. Number
PLC Programmable Logic Controller
  • References
  • SOP for Qualification of Equipments and Utilities
  • Validation Master Plan
  • SOP No. QC224 (Recovery Of Endotoxin From Rubber Stoppers, Vials,       Aluminium Containers And Rubber Bungs)
  • Enclosures
    • Annexure – 1 (Recording Of Observations For Performance Qualification)
    • Annexure – 2 (Performance Qualification Report)
  • REQUALIFICATION CRITERIA

Requalification criteria is as follows

  • If any major changes or modification in the equipment.
  • Change of cycle program
  • Inclusion of new load

                       

Placement Of Endotoxin Indicator Vials And Vials With Temperature Sensors On The Conveyor
Sensor No. Location Justification for location selection
1 and 6 Left side of the conveyor belt. To verify uniform heat penetration in the vials on the left side of the conveyor belt in the tunnel, which is considered to be the critical area where there may be a possibility of cold spot because the hot air re-circulation ducts are below the conveyor belt.
4 and 9 Towards left side from the middle of the conveyor belt. To verify uniform heat penetration in the vials across the conveyor belt towards left side from the middle of the conveyor belt.
2 and 7 In the middle portion of the conveyor belt. To verify uniform heat penetration in the vials across the conveyor belt in the middle portion of the conveyor belt.
5 and 10 Towards right side from the middle of the conveyor belt. To verify uniform heat penetration in the vials across the conveyor belt towards right side from the middle of the conveyor belt.
3 and 8 Right side of the conveyor To verify uniform heat penetration in the vials on the right side of the conveyor belt in the tunnel, which is considered to be the critical area where there may be a possibility of cold spot because the hot air re-circulation ducts are below the conveyor belt.

Note: For Heat distribution study, only temperature sensors shall be considered.

ANNEXURE -1

Details Of Operational Qualification Report
Report No.   Date Of Authorisation  
  Identification Of The Executors
S. No. Name Designation & Department  Sign & Date Training Details
       

Comments:

Measurement of Airflow Velocity

Drying Zone:

Name of the Instrument: Anemometer

Calibration certificate No:______________________ Validity: From:__________To:__________

Calculations:

Minimum Number of measuring locations = 4 locations are decided for measuring velocity.

Acceptance Criteria: Airflow velocity across the HEPA filter should be within the range of 98 ± 20% Feet/Minute.

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 Comments:

Hot Zone:

Name of the Instrument: Anemometer

Calibration certificate No:______________________ Validity: From:__________To:__________

Calculations:

Minimum Number of measuring locations = 4 locations are decided for measuring velocity.

Acceptance Criteria: Airflow velocity across the HEPA filter should be within the range of 138 ± 20% Feet/Minute.

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 Comments:

Cool Zone:

Name of the Instrument: Anemometer

Calibration certificate No:______________________ Validity: From:__________To:__________

Calculations:

Minimum Number of measuring locations = 4 locations are decided for measuring velocity.

Acceptance Criteria: Airflow velocity across the HEPA filter should be within the range of 118 ± 20% Feet/Minute.

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 

Filter No. Filter/ Grill Area (A):
Location Air velocity in Feet/Minute Air Flow

F = V x A

CFM

Checked by Sign / Date
Run 1 Run 2 Run 3 Average Average of all locations
Location 1              
Location 2        
Location 3        
Location 4        

 Comments:

Differential Pressure Test
Photohelic Gauge ID Calibration Details From With respect to Differential pressure in MMWG Limit on display

in mmwc

Checked by

Sign / Date

Certificate No. From To Run – 1 Run – 2 Run – 3
Drying zone Vial washing side Equal to or above 21.2
Hot zone to Drying zone Equal to or above 25
Sterility testing room Change room 2 Equal to or above 25

Comments:

Installed Filter System Integrity and Leakage Test (DOP /PAO Test)

Name of the Instrument: Aerosol Photometer

Calibration Certificate No:______________________Validity: From:__________To:__________

Aerosol used:________________________________

Acceptance Criteria: Designated leaks are deemed to have occurred where a reading greater than 10−4 (0.01%) of the upstream challenge aerosol concentration and should be ‘zero’ through mounting joints. Alternative acceptance criteria may be agreed based on the filter efficiency.

S. No. Filter ID No. Efficiency

 %

Upstream Concentration % Downstream Concentration % Mounting joints

%

Checked by

Sign / Date

Drying zone
1
Hot zone
1
2
Cool zone/Stabilizing zone
1
2
3

 Comments: 

 Airflow Pattern Test

Chemical Used: Titanium Tetra Chloride or Dry Ice

Acceptance criteria:

  • Air should flow from the higher-pressure zone to low pressure zone.
Airflow Pattern/Direction Airflow towards Pre filter

Pass/Fail

Checked by

Sign / Date

From (High pressure zone) To (Low pressure zone) Pass / Fail
Drying zone Vial washing side
Hot / Sterilising zone Drying zone
Cool zone
Vial filling side Cool zone
Non Viable Particle Count Test

 

Stage: Before / After Depyrogenation cycle Vial Size:

Zone Name: Drying Zone                                                          

Name of the Instrument: Laser Based Air Borne Particulate Counter.

Calibration Certificate No:______________________Validity: From: _________To:_________

Test Conditions: At Rest; Specified Class: ‘A’

Calculations:

Number of locations          =       5 locations are decided for sampling

Sample Volume               =       3 cycles per location

Acceptance limit: Maximum permitted number of particles/m3 equal to or above

                                 0.5μm: 3500; 5μm: 20

 

Observations from raw data

S. No. Checks Date Verified by / Date
1 Date of testing    
2 Date of completion    

 

S. No. Checks Observation

Yes / No

Verified by / Date
1 Number of cycles per location is met with the above calculations.    
2 Number of sampling locations is met with the above calculations.    
3 Each cycle of each location met with the acceptance criteria    
4 Average of each location is met with the acceptance criteria    
5 Mean of all locations is met with the acceptance criteria    
6 95% UCL is complies as per the acceptance criteria.    

Zone Name: Hot/Sterilising Zone                                          

Name of the Instrument: Laser Based Air Borne Particulate Counter.

Calibration Certificate No:______________________Validity: From: _________To:_________

Test Conditions: At Rest; Specified Class: ‘A’

Calculations:

Number of locations          =       7 locations are decided for sampling

Sample Volume               =       3 cycles per location

Acceptance limit: Maximum permitted number of particles/m3 equal to or above

                                 0.5μm: 3500; 5μm: 20 

Observations from raw data

S. No. Checks Date Verified by / Date
1 Date of testing    
2 Date of completion    

 

S. No. Checks Observation

Yes / No

Verified by / Date
1 Number of cycles per location is met with the above calculations.    
2 Number of sampling locations is met with the above calculations.    
3 Each cycle of each location met with the acceptance criteria    
4 Average of each location is met with the acceptance criteria    
5 Mean of all locations is met with the acceptance criteria    
6 95% UCL is complies as per the acceptance criteria.    

Zone Name: Cool Zone                                      

Name of the Instrument: Laser Based Air Borne Particulate Counter.

Calibration Certificate No:______________________Validity: From: _________To:_________

Test Conditions: At Rest; Specified Class: ‘A’

Calculations:

Number of locations          =       15 locations are decided for sampling

Sample Volume               =       3 cycles per location

Acceptance limit: Maximum permitted number of particles/m3 equal to or above

                                 0.5μm: 3500; 5μm: 20 

Observations from raw data

S. No. Checks Date Verified by / Date
1 Date of testing    
2 Date of completion    

 

S. No. Checks Observation

Yes / No

Verified by / Date
1 Number of cycles per location is met with the above calculations.    
2 Number of sampling locations is met with the above calculations.    
3 Each cycle of each location met with the acceptance criteria    
4 Average of each location is met with the acceptance criteria    
5 Mean of all locations is met with the acceptance criteria    
 Heat Distribution Study For Empty Chamber

 

DEPYROGENATION CYCLE PARAMETERS
Parameter
PLC set point
Parameter
PLC set point
Set point (deg C) HZ over shoot (deg C)
Conveyor start temperature (deg C) Conveyor stop temperature (deg C)
Conveyor speed (mm/min) CZ over shoot (deg C)
Data logging time (min) Night mode HZ temp (deg C)
Recipe type

 

Observations from Print outs:

S. No. Checks Run – 1 Run – 2 Run – 3 Verified by Sign & Date
1 Date of testing        
2 Date of completion        
3 Done by        

 

S. No. Checks Observation Yes / No Verified by Sign & Date
Run – 1 Run – 2 Run – 3
1 The temperature at each temperature sensor should be within ± 250C during the depyrogenation cycle.        
2 The temperature distribution is uniform throughout the sterilization period i.e. ± 250C to set temperature for at least 3.0 min.        
3 Conveyor speed (mm/Min) is observed as per the set point through out the cycle.        
4 Non viable particle count has taken after validation runs and complies the acceptance criteria.        
 Heat Penetration Study

 

DEPYROGENATION CYCLE PARAMETERS
Parameter
PLC set point
Parameter
PLC set point
Set point (deg C) HZ over shoot (deg C)
Conveyor start temperature (deg C) Conveyor stop temperature (deg C)
Conveyor speed (mm/min) CZ over shoot (deg C)
Data logging time (min) Night mode HZ temp (deg C)
Recipe type

 

Observations from Print outs:

S. No. Checks Run – 1 Run – 2 Run – 3 Verified by Sign & Date
1 Vial type/size        
2 Date of testing        
3 Date of completion        
4 Done by        

 

S. No. Checks Observation Yes / No Verified by Sign & Date
Run – 1 Run – 2 Run – 3
1 Conveyor is packed with vials during run.        
2 Is vial is moved on the conveyor belt in erect condition. No vial should be tilted.        
3 Any breakage of vial observed during run.        
4 The temperature at each temperature sensor is within ± 250C during the depyrogenation cycle.        
5 The temperature distribution is uniform throughout the sterilization period i.e. ± 250C to set temperature for at least 3.0 min.        
6 Conveyor speed (mm/min) is observed as per the set point through out the cycle.        
7 Exposed endotoxin indicators are meeting the minimum 3-log reduction        
8 Non viable particle count has taken after validation runs and complies the acceptance criteria.        

Discrepancy And Corrective Action Report

 

ANNEXURE -2

Heat Penetration Study For Load Chamber

 

Spiking of Endotoxin Vials

Vial Type/ Size:                                                              

Endotoxin Indicators
Lot No. Amount
Expiry Date Reconstitution on
LRW Details
LRW Lot No.: Expiry Date:
Date of Opening:
Spiking details
Micropipette ID: LAF ID:
Validation CSE Potency: Reconstitution:
Spiked volume: Number of Spiked vials:
Done By:

  

 Recovery of Spiked Endotoxin

 Confirmation Of Labelled Claim Sensitivity Of The Lysate

LRW Details LAL Details Control Standard Endotoxin Details
LRW Lot No. LAL Lot No. Lot No.
Expiry Date Sensitivity Expiry Date
Date of Opening Date of Reconstitution Conc.
Expiry Date Reconstituted on
Endotoxin Indicators
Lot No. Amount
Expiry Date Reconstitution on

Control Standard Series

Tube No. LRW ( ml) Vol of CSE added from Tube Final CSE Concentration
01      
02      
03      
04      
05      
06      

           Observation:

           Time of Incubation          :                                   Time of Observation           :

  Temp. at Incubation        :                                   Temp. at Observation        :

Replicate No. Observation at different concentrations End Point Log 10

End Point

– ve Control

(0.1 ml LRW + 0.1 ml lysate)

2 λ λ λ/2 λ/4
I              
II              
Mean of log  10 End Point      
Geometric Mean = Antilog 10 of Mean End Point    

Remarks:  The geometric Mean End Point concentration is within the / Not within the ± two fold of the labelled claim sensitivity of the lysate.

Done By:

RECOVERY OF ENDOTOXIN IN UNEXPOSED SPIKED VIALS

No. of Unexposed vial used for determining the recovery        =  01

Endotoxin Concentration Spiked on each vial                        = 10000 EU /vial

Volume of LRW required for dilution                                    = 4.0 ml

Total concentration in vial will be 10000/4                           =2500 EU/ ml

Dilutions:

Tube No. Volume of LRW added Volume of Endotoxin Concentrations Final Concentration Dilution
0.9 ml 0.1 ml of 2500 EU/ml 250 EU /ml 1: 40
0.9 ml 0.1 ml of Tube No.1 25 EU / ml 1: 400
0.9 ml 0.1 ml of Tube No.2 2.5 EU / ml 1: 4000
1.8 ml 0.2 ml of Tube No.3 0.25 EU / ml 1: 40000
1.0 ml 1.0 ml of Tube No.4 0.125 EU / ml 1: 80000
1.0 ml 1.0 ml of Tube No.5 0.06 EU / ml 1: 160000

Observation:

           Time of Incubation:                                                Time of Observation:

           Temp. at Incubation:                                          Temp. at Observation:

Dilution Sample Vol. of Sample Vol. of Lysate Result
·        – ve Control Test 100 µl of LRW 100 µl
Duplicate 100 µl of LRW 100 µl
·        1 : 40 000

 

Test 100 µl 100 µl
Duplicate 100 µl 100 µl
·        1 : 80 000

 

Test 100 µl 100 µl
Duplicate 100 µl 100 µl
·        1 : 160000

 

Test 100 µl 100 µl
Duplicate 100 µl 100 µl

ENDOTOXIN CONTENT IN EXPOSED VIALS AFTER DEPYROGENATION

  • Concentration of Endotoxin Spiked on each vial = 10000 EU / vial.
  • Assuming that after depyrogenation a three log reduction must have taken place, the total concentration on each vial will be = 10 EU / vial
  • Volume of LRW used for Dilution = 4.0 ml / vial
  • Therefore the total concentration in vial will be = 2.5 EU / ml

      Dilutions:

Tube No. Volume of LRW added Volume of Endotoxin Concentrations Final Concentration Dilution
1. 1.8 ml of LRW 0.2 ml of 2.5 EU / ml 0.25 EU /ml 1: 40
2. 1.0 ml of LRW 1.0 ml of 0.25 EU / ml 0.125 EU /ml 1: 80

 

Dilution Sample Vol. of Sample Vol. of Lysate
·        1 : 40 Test 100 µl 100 µl
·        1 : 40 Duplicate 100 µl 100 µl
·        1 : 80 Test 100 µl 100 µl
·        1 : 80 Duplicate 100 µl 100 µl

            Observation:

           Time of Incubation :                                               Time of Observation           :

           Temp. at Incubation           :                                   Temp. at Observation        : 

Vial No. Dilution  1: 40 Dilution 1: 80 Vial No. Dilution 1: 40 Dilution 1: 80
Test Duplicate Test Duplicate Test Duplicate Test Duplicate

 

ESTIMATION OF ENDOTOXIN REDUCTION FOR RUN NO._______

  1. Recovered endotoxin per             = Reciprocal of the last dilution which

           Exposed vials           was positive X lambda (Sensitivity of the Lysate).

  • Last Dilution which was positive =…………….
  • Therefore the reciprocal will be         =…………….
  • Sensitivity of the Lysate = 0.125 EU / ml

=  …………….  X 0.125 EU/ ml

=  ……………….. EU / vial

Recovered endotoxin per            = Reciprocal of the last dilution which

Unexposed vial          was positive X lambda (Sensitivity of the Lysate).

  • Last Dilution which was positive =…………….
  • Therefore the reciprocal will be         =…………….
  • Sensitivity of the Lysate = 0.125 EU / ml

=   ……………….  X 0.125 EU/ ml

=  ……………….. EU / vial

therefore the amount of endotoxin reduced after depyrogenation

= log (b) –  log (a)

= log …………-  log ……. ….

=  ……………….log reduction

Anti log of (log reduction)              =   ……………….fold reduction

  • INTERPRETATION:

There is greater / less than 3 log reduction in endotoxin in Depyrogenated vial. Hence depyrogenation cycle was successful / not successful.

 

Reviewed by:                                                                                            Date:

 

 

 

 

 

About Pharmaceutical Guidanace

Mr. Shiv Kumar is the Author and founder of pharmaceutical guidance, he is a pharmaceutical Professional from India having more than 14 years of rich experience in pharmaceutical field. During his career, he work in quality assurance department with multinational company’s i.e Zydus Cadila Ltd, Unichem Laboratories Ltd, Indoco remedies Ltd, Panacea Biotec Ltd, Nectar life Science Ltd. During his experience, he face may regulatory Audit i.e. USFDA, MHRA, ANVISA, MCC, TGA, EU –GMP, WHO –Geneva, ISO 9001-2008 and many ROW Regularities Audit i.e.Uganda,Kenya, Tanzania, Zimbabwe. He is currently leading a regulatory pharmaceutical company as a head Quality. You can join him by Email, Facebook, Google+, Twitter and YouTube

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