Protocol for  Validation of Sterilization Process using Steam Sterilizer (Bung Processor)

1
2159

Protocol

For

Validation Of Sterilization Process

Using

Steam Sterilizer (Bung Processor)

Equipment ID

Aseptic Processing Area

Location: Pharmaceutical Company

Document No.
Supersedes
Effective Date
No. of Pages

 

PROTOCOL CONTENTS

Sr. No.                    Section Title                                                                              Page No.

1.0       Protocol Approval……………………………………..…………

2.0       Overview………………………………………………………….

                  2.1    Objective……………………………………………………

                  2.2    Purpose and Scope…………………………………………

                  2.3    Responsibility……………………………………………….

                  2.4    Validation Team………………………………………….…

3.0         Pre-Validation Requirement…………………….………………

                  3.1    Pre-Requisite…………………………………………….….

                  3.2    Verification of Instruments Calibration………………….

                  3.3    Preventive Maintenance Status.……………………….….

                  3.4    Steam Quality Tests…………………………………….….

4.0         System/Equipment Description..……………….………………

                  4.1    System/Equipment Details…………………………….…..

                  4.2    Design………………………………………………….…..

5.0         Validation Procedure …………………….……………………..

                  5.1    Flow Charts……………..…………………………….……

                  5.2    Testing Parameters……………..……………….…………..

6.0         Acceptance Criteria …………………….………………………..

7.0         Validation Report…………………………………………..….

8.0         Approval of Validation Report……………………………..….

9.0         Observed Deviation…………………………………….….…..

10.0       List of Exhibits/Annexure……………………………………….

11.0       Reference Documents..…………… ……………………………

12.0        Periodic validation……………………………………………. 

13.0        Revision history…………………………………………………

 

1.0 Protocol Approval:

 

Department Name Designation Sign & Date
Prepared by
Production Injection
Quality Assurance

Reviewed by

HOD – Production
HOD – Microbiology
HOD- Engineering
Quality Assurance
Approved by
HOD –Quality Assurance

 

2.0 Overview:

The container closure components of a sterile drug product are required to be sterilized in aseptic processing. The Sterilization process using Steam Sterilizer (Bung Processor) is followed for porous and nonporous load. The autoclave sterilization process helps in inactivation of Biological indicators to a log reduction value more than 6. This process of sterilization of different loads in different pattern is required to be validated as per GMP requirement.

2.1 Objective:

The objective of this protocol is to describe the procedure and plan for validation of sterilization process of porous and non porous load using Steam Sterilizer (Bung Processor)

2.2 Purpose:

The purpose of this protocol is to help the operating personnel to plan the validation activities so as to demonstrate that the Steam Sterilizer (Bung processor) operates consistently to inactivate the Biological indicators from the Loads to more than 6 log reduction and thus provide a high degree of assurance.

2.3 Scope:

The scope of this protocol is Limited to the validation of sterilization process for all types of Loads used in pharmaceutical manufacturing Unit , using the Steam Sterilizer (Bung processor) Make Machine Fabric.

2.4 Responsibility:

2.4.1 The validation team shall be responsible for the execution of the validation activity.

2.4.2 Engineering department shall be responsible for the support to the executing team.

2.4.3 QC shall be responsible for the testing of the validation samples and compilation of results.

2.4.4 Head –QA shall be responsible for the compliance of this protocol.

2.5 Validation Team:

The group comprises of representatives from each of the following departments and they shall be responsible for the overall compliance as per this protocol.

Department Responsibilities
Engineering & Utility Arranging for execution and recording of the following activities
· Calibration of the gauges and instruments· Steam Quality Test· Vacuum Leak Test· Periodic Calibration of Data logger during Performance qualification· Heat distribution studies.· Heat penetration studies.· Vacuum break filter integrity testing.· Review of Protocol and Report
Production · Execution and collection of the data.· Preparation and Review of Protocol and Report
Quality Control · Testing of the samples during the study to Conduct chemical, Microbiological testing etc., as per the test procedures.· Review of Protocol and Report
Quality Assurance · Review and approval of Protocol and Report· Verifying the compliance of existing procedures to theoutcome of validation

 

3.0 Pre-Validation Requirements:

3.1 Pre-Requisite:

Following instruments and materials shall be required for the Validation of Bung Processor Cum Steam Sterilizer of dry powder injection production area.

  • Calibrated Data logger
  • Calibrated PT 100, RTD probes.
  • Bowie-Dick Test packs
  • Geobacillus stearothermophilus ATCC 7953 e. Biological Indicator strips
  • Rubber Bungs
  • Endotoxin spiked bungs
  • Loads for Validation (porous, Non-porous and Liquid)

The HPHV Steam Sterilizer shall be validated after verification of the documented evidence (obtained as per the methods outlined in this Protocol) assuring that the equipment is meeting the desired performance attributes repeatable & reproducible consistently.

The following pre-validation test shall be performed:

S. No. Test Parameters Details
1 Verification of Calibration · Before start of the validation activity.
2 Steam Quality Tests · To be performed as per respective SOP.
3 Vacuum Leak Test ·Empty Chamber·Temperature Probes Connected.-Without Temperature Probes.
4 Bowie –Dick Test ·    3 consecutive Runs on 3 different days
5 Empty Chamber Heat Distribution study With temperature mapping probes at different locations of the sterilizer chamber.

To qualify these tests the equipment should fulfill the acceptance criteria as described. After completion of the validation tests all the data generated will be compiled together to evaluate capability of the HPHV Steam Sterilizer.

 3.2 Verification of Instruments Calibration:

The Calibration of Gauges and Instruments shall be carried out prior to validation activity and recorded in the table as given below.

Instrument Range I.D. No. Location Calibrated Date Next Due Date
Pressure Gauges 0 to 6 kg/cm² Jacket
0 to 10  kg/cm² Pure Steam
0 to 10  kg/cm² Compressed Air
0 to 16 Bar Air Regulator
0 to 10  kg/cm² WFI Line
0 to 10  kg/cm² Compressed Air
0 to 10  kg/cm² Sampling point
0 to 10  kg/cm² Filter
0 to 10 Bar For F.R. Assembly
0 to 10 Bar For Door Gasket
0 to 10 Bar For Door Cycle
0 to 10  kg/cm² Steam Line
Compound Gauges -1 to 6  kg/cm² Chamber
-1 to 6  kg/cm² Non Sterile Gasket
-1 to 6  kg/cm² Sterile Gasket
-1 to 6  kg/cm² Sterile
-1 to 6  kg/cm² Chamber
Vacuum Gauge 0 to -760mmHg Vacuum Line
Temperature Sensors 0 to 250 °C Chamber
0 to 250 °C Chamber
0 to 250 °C Chamber
0 to 250 °C Chamber
0 to 250 °C Drain
Data Logger      
RTD Sensors.      

 

3.3 Preventive Maintenance status:

The Preventive Maintenance of Steam sterilizer and the support systems shall be carried out before the validation activity and recorded as per the table given below:

Equipment I.D. No. Location Preventive Maintenance  date Next Due Date
Steam Sterilizer (Bung Processor)
Pure steam
Water For Injection
Compressed air
Purified water
Data logger

 

3.4 Steam Quality Tests:

The quality of the steam shall be tested for the content of Non-Condensable Gases, Superheat and Steam Dryness Value, on three different days (three trials) as per respective SOP and record the results in Format.

Acceptance Criteria

Non-condensable gases: Not more than 3.5 %,

Superheat: Not more than 25°C.

Steam Dryness Value: Not less than 0.90 (if metal loads are to be processed, the dryness value should not be less than 0.95).

4.0 System / Equipment Description:

4.1 System / Equipment Details:

4.1.1 Equipment Description:

Equipment Name  High Pressure High Vacuum Steam Sterilizer (Bung Processor )
Equipment ID
Capacity 1765 Liters
Location Equipment Washing  and Sterilization  Rooms
Manufacturer’s Name M/s Machin Fabrik
Dimension
Height 1050 mm
Width 1050 mm
Depth 1600 mm

 

4.1.2 Working Principle:

4.1.2.1 The steam sterilizer is supplied with pure (clean) steam for the chamber and plant steam to the jacket.

4.1.2.2 The Sterilization cycle can be automatically controlled by the PLC, which can be programmed and protected with three different levels of passwords.

4.1.2.3 The steam sterilizer has the capability to handle the following types of cycles automatically.

  • Vacuum Leak Test.

This cycle is used to assure the sterilizer chamber integrity towards leakage.

  • Bowie-Dick Test.

This cycle is used to ensure steam penetration in to the packs is appropriate.

  • Standard Cycle.

Gravity displacement steam sterilization cycle, suitable for liquid loads.

  • HPHV Cycle.

High Pressure High Vacuum sterilization cycle with vacuum pulses, suitable for porous loads.

All the above cycles are PLC controlled and runs automatically when selected.

4.1.3 Operating cycle for a porous load.

            The selected operating cycle of a porous load sterilizer normally have five stages.

  • Air Removal – Sufficient air is removed from the chamber and the load to permit attainment of the sterilization conditions.
  • Steam Admission – Steam is admitted to the chamber until the specified sterilization temperature is attained throughout the chamber and load.
  • Holding Time – The temperature throughout the chamber and load is maintained within the sterilization temperature band for the appropriate holding time.
  • Drying – Steam is removed from the chamber and the chamber pressure is reduced to permit the evaporation of condensate from the load by prolonged evacuation.
  • Air Admission – Air is admitted to the chamber until the chamber pressure approaches atmospheric pressure.

4.1.4 Area Description

  • The HPHV Steam Sterilizer is located in the Preparation and sterilization room of the sterile powder injectable filling section with restricted access.
  • The Preparation and sterilization room is designed as per the specifications for cleanliness Grade – D.
  • The equipment is located such that, it can be attended easily for routine operational, monitoring and maintenance purpose.
  • One door of the sterilizer opens into the Preparation and sterilization room for loading and the other door opens into the aseptic area for unloading.
  • The control panel is located near to the equipment (in Grade – D) for ease of operation.
  • The other supporting systems such as, water-ring vacuum pump is located near the sterilizer and the pure steam generator is located in the area dedicated for water system. Compressed air supply is from centralized system.

4.2 Design:

  • The Steam Sterilizer (Bung Processor) consists of the following features.
  • The Sterilization Chamber is made up of SS sheet, which is welded with U-Profile SS Jacket.
  • The Sterilization Chamber is provided with two sliding doors, which are made up of SS reinforced with MS Support structure.
  • The door is operated with the help of pneumatic cylinder.
  • The door seals 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.
  • When the door reaches the end position (closed) the gaskets are pushed out automatically with the help of compressed air for leak proof sealing.
  • Similarly to open the door, gasket is retracted with the help of vacuum, which is created by the compressed air ejectors.
  • When the gasket is retracted the sterilizer chamber door slides automatically.
  • Door interlocks are provided to prevent simultaneous opening of both the doors and process lock to prevent opening of the door during the operation.
  • The sterilization 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 SS cladding
  • A Stainless Steel pipe stand support is provided for the equipment thus requiring no special foundation.
  • To ensure leak-tight partition between the aseptic area (Unloading side) and washing & sterilization area (Loading side) an SS flush-panel is provided on the partition wall and the outer cover of the Bung Processor. All Joints, Crevices are filled with Epoxy/Silicon sealant to prevent possible leakage.
  • The Bung Processor is provided with the following systems and accessories for the desired functioning.
  • Water-ring type vacuum pump with suitable electric motor. (SS shell and tube type condenser for condensation of steam prior to entering the housing of the vacuum pump).
  • Sterilizing grade vacuum-break filter on the sterile area (unloading side), which ensures sterility of the load by introducing sterile filtered air after vacuum break.
  • Two numbers of removable SS railings inside the sterilization chamber for smooth and easy operation of the carriage.
  • One stainless steel (SS) rotating carriage with rubber stopper holding cassettes having four SS Constructed of SS with four SS wheels with sealed bearings the rotating carriage is driven by an in-built reduction-gear mechanism during rubber stopper sterilization and vacuum drying cycle. Rubber stoppers can be washed, siliconised, Steam Sterilized and vacuum dried with the aid of this mechanism.
  • Two floor trolleys (one each in loading and unloading side), constructed of SS with four swiveling castor wheels made of polypropylene. Each trolley is suitable for loading rotating carriage and situated one in aseptic and another in non-aseptic areas.
  • Another chamber trolley with shelves is provided to handle different types of loads in sterilization chamber.
  • Chamber is provided with compound gauges on the sterile area side (Unloading side) and the preparation & sterilization area side (Loading side).
  • Jacket pressure gauge, Gasket pressure gauge for both the doors, Safety valve for jacket and chamber, Steam trap with strainer and NRV for chamber and steam trap for jacket, on the washing & sterilization area side (Loading side).
  • Control panel consisting of PLC, Digital Temperature input to the microprocessor from a 5 numbers of resistance temperature detectors (RTDs), one number of Pressure transmitter, pressure and vacuum switches for transmitting digital Pressure/Vacuum signals to the microprocessor, provision of auto-override for manual operation in case of microprocessor failure.
  • SS Piping, Ball valves with rotary actuators for all the process lines and solenoid valves to regulate air supply to the pneumatic rotary actuators.

5.0 Validation Procedure:

5.1 Flow Charts:

Steam Sterilizer (Bung Processor) Validation Process Flow Chart

Steam-SterilizerProcess Flow Chart for Heat Penetration Study of Non-RFS Rubber Bungs 

Process-Flow-Chart-for-Heat-Penetration-Study-of-Non

5.2 Testing Parameters:

5.2.1 Vacuum Leak Test

The vacuum leak test shall be performed as per the SOP on “Operation of Vacuum leak test in bung processor”.

  • Empty Chamber
  • Temperature Probe Connected
  • Without Temperature probe connected

The observations and results shall be recorded in exhibit enclosed as Exhibit-E01.

Acceptance Criteria

Vacuum Leak Rate – NMT 0.013 Bar / 10 minutes.

5.2.2 Bowie Dick Test:

Follow respective SOP to perform the Bowie Dick Test during the study. Record the observations and results in formats enclosed as Exhibit-E02.

5.2.3 Bowie-Dick Test Papers

  • The Bowie-Dick test uses a Class B chemical indicator contained within a standard test pack.
  • The indicator is distributed over an A4 paper sheet in the form of a geometric pattern.
  • This indicator paper is placed at the centre of a test pack of folded huckaback towels and then subjected to an operating cycle. The indicator tape shows a change of color in response to a combination of time, temperature and moisture.
  • When used in conjunction with a standard test pack, Class B indicators are designed to show a failure either if, at the start of the holding time, the temperature at the centre of the test pack is 2°C or more below the temperature in the active chamber discharge; or if the indicator is exposed to insufficient moisture.
  • Both conditions are usually caused by the presence of air or other non-condensable gases

Acceptance Criteria

  • There should be a uniform colour change throughout the indicator.
  • No change, non-uniform change and/ or air entrapment (bubble) spot on the   Pattern indicates inadequate air removal from the sterilization chamber.

5.2.3 Heat Distribution Empty Chamber Study

  • Record the PLC parameters set for the sterilization cycle during empty chamber heat distribution study, in the formats of Exhibit-E03.
  • Place the 16 Temperature mapping probes into chamber through the port of the sterilizer. Seal the port with silicone sealant so that steam leakage does not take place.
  • Suspend the probes in the chamber in different position so that probes do not touch any metallic surface.
  • Connect the probes to a calibrated data logger, which can scan and print the actual temperature observed at different locations with respect to time.
  • Collect sufficient sample of steam condensate as per the SOP on monitoring of purified water, water for injection and pure steam condensate respective SOP for analysis in a clean and sterile sample bottle from the sampling point provided in the steam condensate drain line.
  • Select HPHV sterilization cycle on control panel. Operate the Steam Sterilizer as per respective SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time.
  • When the sterilization cycle is complete, collect Strip chart from the Strip chart Recorder of the Steam Sterilizer and enclose to Exhibit-E03
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to Exhibit-E03.

Cold Point: The sample location that is the slowest to heat amongst the total number of sample locations is called as Cold Point.The following table describes placement of External Temperature probes placement in the Empty Chamber and justification for selection of locations :

  • Probe Location in the Chamber Justification for location selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    EP 2 Steam inlet right To verify the probability of excess temperature near the steam inlet.
    EP 3 Steam inlet left
    EP 4 Sterile side door top back side Any conduction of heat through the door, which may cause temperature drop at that particular point.
    EP 5 Sterile side door bottom back
    EP 6 Non-aseptic door top front side
    EP 7 Non-aseptic door bottom front side
    EP 8 Near RTD one As these are temperature recording probes at different points of sterilizer. It is necessary to verify/compare the temperatures obtained with strip chart / print out from the sterilizer.
    EP 9 Near RTD two
    EP 10 Near RTD four
    EP 11 Near RTD three
    EP 12 Near RTD five
    EP 13 Non-aseptic door top Left corner To know the temperature distribution in these regions.
    EP 14 Non-aseptic door right corner
    EP 15 Aseptic door top left corner
    EP 16 Aseptic door bottom corner
    (NOTE: The temperature probe shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels).

     

    Inbuilt Temperature Probes Location
    IP 1 Drain
    IP 2 Right Rear
    IP 3 Left Front
    IP 4 Left rear
    IP 5 Right Front

     Numbering and location of Internal inbuilt probe in the chamber

Internal & External Temperature probes placement in the Empty Chamber Study:

1

The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistency.

If the results obtained from the empty heat distribution study are satisfactory, perform (repeat) two more times to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.

Compile the data generated during the qualification study for complete evaluation of the sterilization process in the empty chamber. The data compiled from the three

runs shall be reviewed and a report of the heat distribution study shall be prepared and approved.

 5.2.4 Heat Penetration Studies

The following validation test shall be performed in the Loaded Chamber heat penetration studies.

The following are the fixed load patterns selected for qualification of the sterilizer

S.No. Load/Test Details
1 –         Aseptic area Garments-         Filling machine parts-         Filtration accessories-         Rubber stoppers in RFS bags-         Rubber stoppers after washing ·     Temperature mapping probes shall be placed inside the innermost part (assumed to be difficult to attain sterilization temperature i.e. cold spot) of the load.
·     Bio-Challenge studies shall use Geobacillus stearothermophilus  spore strips (containing more than 10 6  spores per strip) during the heat penetration studies.
·     Estimation of the FO value achieved during the sterilization hold period at each temperature-mapping probe.
2 Liquid nutrient media for aseptic media fill. ·  Temperature mapping probes shall be placed inside the liquid vessel to ensure the hold time. Estimation of the FO value achieved during the sterilization hold period at each temperature-mapping probe
3 Steam Quality Tests ·     Quality of the steam condensate collected during the validation of fixed loads shall be tested.

Sterilization cycle time

  • The sterilization cycle time of 30 minutes at 1210C sterilization temperature is selected. This is the time selected set on the PLC to carry out the sterilization cycle automatically.
  • A worst-case bioburden contamination of more than 106 heat resistant spore-forming bacteria (Geobacillus stearothermophilus ATCC 7953) is considered.
  • The worst-case D-value of 2.5 minutes is considered, which is normally around 2.0 minutes.
  • Hence to achieve a Sterility assurance level of 10-6 (12 log reduction) the length of an overkill sterilization cycle at 1210C will be 30 minutes (12 log X 2.5 min/log= 30 minutes).

Heat Penetration Study for Garment Load

  • The load articles shall not be placed near the cold spot / point as this is the slowest to heat.
  • 03 numbers of cycles shall be considered for the maximum and minimum Loading pattern of garments. For operation of Steam Sterilizer respective SOP , “Operation of HPHV steam sterilizer (Bung processor).
  • Record the PLC set parameters same for both minimum and maximum load configuration described for the sterilization cycle during loaded chamber heat penetration study in the Exhibit-E04 and Exhibit-E05
  • Place 16 Temperature mapping probes into chamber through the port of the sterilizer. Seal the port with silicone sealant so that steam leakage does not take place.
  • Place the probes and BI inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement for garment minimum and. for garment maximum loads.
  • Also place biological indicator strips along with each temperature-mapping probe.
  • Connect the probes to suitable data logger, which can scan and print the actual temperature with respect to time.
  • Operate the steam sterilizer as per respective SOP, and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. Time set on PLC, Strip chart recorder, Printout and Data Logger must correspond to each other.
  • Record the results of the steam condensate in the respective Exhibits.
  • When the steam sterilization cycle is complete, collect Strip chart/ printout of the steam sterilizer and attach to Exhibit-E04 and Exhibit-E05,
    • Download the data from data logger into the computer for data-analysis and printing. Attach the data printouts to the Exhibit-E04 and Exhibit-E05,
    • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology laboratory.
    • Location and Justification for Temperature probes Placement (Garment Load Minimum)
    Probe  No. Location in the Chamber Justification for Location Selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    Top rack of the trolley.
    EP 2 Inside Garment pack -1 To verify the temperature penetration in the inner most portion of the pack.
    EP 3 Inside Garment pack -2
    EP 4 Inside Garment pack -3
    Middle rack of the trolley.
    EP 5 Inside Garment pack -4 To verify the temperature penetration in the inner most portion of the pack. Loading level is middle of the trolley.
    EP 6 Inside Garment pack -5
    EP 7 Inside Garment pack -6
    Bottom rack of the trolley.
    EP 8 Inside Garment pack -7 To verify the temperature penetration in the inner most portion of the pack. Loading level is bottom of the trolley.
    EP 9 Inside Garment pack -8
    EP 10 Inside Garment pack -9
    EP 11 50 mm above outside the Garment pack -3  towards aseptic door To verify the temperature distribution inside the chamber in these locations during sterilization process.
    EP 12 50 mm above outside the Garment pack -3 towards aseptic door
    EP 13 50 mm above outside the Garment pack -4 towards non-aseptic door
    EP 14 50 mm above outside the Garment pack -6 towards aseptic door
    EP 15 50 mm above outside the Garment pack -7 towards non- aseptic door
    EP 16 50 mm above outside the Garment pack -9 towards aseptic door

    Note: The temperature probes shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels.

    • Temperature probe Placement (Garment Load Minimum)2
      • Temperature probe Placement (Garment Load Maximum)
      Probe  No. Location in the Chamber Justification for Location Selection
      EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
      Top rack of the trolley.
      EP 2 Inside Garment pack -1 To verify the temperature penetration in the inner most portion of the pack.
      EP 3 Inside Garment pack -3
      EP 4 Inside Garment pack -5
      EP 5 Inside Garment pack -7
      Middle rack of the trolley.
      EP 6 Inside Garment pack -9 To verify the temperature penetration in the inner most portion of the pack.
      EP 7 Inside Garment pack -11
      EP 8 Inside Garment pack -13
      EP 9 Inside Garment pack -15
      Bottom rack of the trolley.
      EP 10 Inside Garment pack -17 To verify the temperature penetration in the inner most portion of the pack.
      EP 11 Inside Garment pack – 19
      EP 12 Inside Garment pack – 21
      EP 13 Inside Garment pack – 23
      EP 14 50 mm above outside the Garment pack -5 towards non aseptic door To verify the temperature distribution inside the chamber in these locations during sterilization process.
      EP 15 50 mm above outside the Garment pack -11 in the middle of chamber aseptic door
      EP 16 50 mm above outside the Garment pack -20 towards aseptic door

      Note: The temperature probes shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels. The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistency.

    • Temperature probe Placement (Garment Load Maximum)3
      • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load (both minimum and maximum) to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
      • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the garment loads. The data compiled from the three runs each load type shall be reviewed and a report of the heat penetration study shall be prepared and approved.

       

      • Heat Penetration Study Rubber Stoppers in RFS Bags:
        • For 20mm and 32mm rubber stoppers Loading pattern and Set parameters refer sop on sterilization of 20 mm and 32 mm RFS rubber stoppers respective SOP.
        • Record the PLC set parameters for the sterilization cycle during loaded chamber heat penetration study in the formats of Exhibit-E06 and Exhibit-E07 for 20 mm and 32 mm rubber stoppers respectively.
        • Connect the probes to a calibrated data logger, which can scan and print the actual temperature observed at different locations with respect to time.
        • Place the probes and BI inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement.
        • When the sterilization cycle is complete, collect Strip chart/ printout of the sterilizer and attach to Exhibits.
        • Download the data from data logger into the computer for data-analysis and printing. Attach the data printouts to Exhibits
        • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology laboratory. The observations shall be recorded in the respective recording sheets.

       

      • Temperature probe placement and justification for location of selection for 20 mm rubber stoppers
      Probe  No. Location In The Chamber Justification For Location Selection
      EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
      In the top rack
      EP 2 Inside pack –  1 To verify the temperature penetration in the inner most portion of the pack.
      EP 3 Inside pack –  2
      EP 4 Inside pack –  3
      In the middle rack
      EP 5 Inside pack –  4 To verify the temperature penetration in the inner most portion of the pack.
      EP 6 Inside pack –  5
      In the bottom rack
      EP 7 Inside pack –  6 To verify the temperature penetration in the inner most portion of the pack.
      EP 8 Inside pack –  7
      EP 9 Inside pack –  8
      EP 10 50 mm above, outside the pack -1 towards non -aseptic door To verify the temperature distribution above the packs.
      EP 11 50 mm above, outside the pack – 3 towards aseptic door
      EP 12 50 mm above, outside the pack – 4 towards non- aseptic door
      EP 13 50 mm above, outside the pack – 5 towards aseptic door
      EP 14 50 mm above, outside the pack – 6 towards non- aseptic door
      EP 15 50 mm above, outside the pack – 7 in middle of chamber
      EP 16 50 mm above, outside the pack – 8 towards aseptic door

      (Note: The temperature probes shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels).          

      • Temperature probe placement for 20 mm rubber stoppers:

4

  • Temperature probe placement and justification for location of selection for 32 mm rubber stoppers
Probe No. Location In The Chamber Justification For Location Selection
EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
In the top rack
EP 2 Inside pack – 1 To verify the temperature penetration in the innermost portion of these packs.
EP 3 Inside pack – 3
EP 4 Inside pack – 4
EP 5 Inside pack – 6
In the middle rack
EP 6 Inside pack – 7 To verify the temperature penetration in the innermost portion of these packs.
EP 7 Inside pack – 9
EP 8 Inside pack – 11
EP 9 Inside pack – 13
In the bottom rack
EP 10 Inside pack – 14 To verify the temperature penetration in the innermost portion of these packs.
EP 11 Inside pack – 16
EP 12 Inside pack – 18
EP 13 Inside pack – 20
EP 14 50 mm above, outside the pack – 1 near aseptic door To verify the temperature distribution above the packs
EP 15 50 mm above, outside the pack – 12 in middle of chamber
EP 16 50 mm above, outside the pack – 20 near non-aseptic door

(Note: The temperature probe shall be placed in the predetermined locations with pre-identified probe numbers corresponding to the data logger channels).

 Temperature probe placement for 32 mm rubber stoppers:

5

  • The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistency of temperature between them
    • Record the observations and results in formats enclosed as Exhibit-E06 for 20 mm rubber stoppers in RFS bags and 32 mm rubber stoppers in RFS bags Exhibit-E07.
    • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the rubber stoppers. The data compiled from the three runs each load shall be reviewed and a report of the heat penetration study shall be prepared and approved.

5.2.4.3 Heat Penetration Study-Sterile Powder Filling Machine Parts:

  • For Loading pattern refer SOP operation of HPHV steam sterilizer (Bung processor)  respective SOP.
  • Record the PLC set parameters for the sterilization cycle during loaded chamber heat penetration study in the formats of Exhibit-E08.
  • Connect the probes to a calibrated data logger, which can scan and print the actual temperature observed at different locations with respect to time.
  • Place the probes and BI inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement.
  • When the sterilization cycle is complete, collect Strip chart/ printout of the sterilizer and attach to Exhibit-E08.
  • Download the data from data logger into the computer for data-analysis and printing. Attach the data printouts to Exhibit-E08.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. The observations shall be recorded in the formats of Exhibit-E08.
  • External Temperature probe placement and justification for location of selection for Sterile Powder Filling Machine Parts Load:
  • Probe No. Location In The Chamber Justification For Location Selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    EP 2 Connector pipe To verify the temperature penetration in the innermost portion of these parts.
    EP 3 Main Powder hopper
    EP 4 Intermediate powder hopper
    EP 5 Bung hopper
    EP 6 Dosing disc
    EP 7 Aspiration sector
    EP 8 Junction L
    EP 9 Junction T
    EP 10 Compressed gas filter
    EP 11 50 mm above the load, in the top rack non- aseptic side To verify the temperature distribution above the machine parts.
    EP 12 50 mm above the load, in the top rack aseptic side
    EP 13 50 mm above the load, in the middle rack aseptic side
    EP 14 50 mm above the load, in the middle rack non-aseptic side
    EP 15 50 mm above the load, in the bottom rack aseptic side
    EP 16 50 mm above the load, in the bottom rack non-aseptic side

    (NOTE: The temperature probes shall be placed in the pre determined locations with pre-identified probes numbers corresponding to the data logger channels). 

    • The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistency of temperatures
    • Record the observations and results in formats enclosed as Exhibit-E08 for Sterile Powder Filling Machine Parts.
    • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the Sterile Powder Filling Machine Parts shall be compiled from the three runs each load shall be reviewed and a report of the heat penetration study shall be prepared and approved

5.2.4.4 Heat Penetration Study for Miscellaneous Items Load:

  • For Loading pattern and set parameter refer SOP operation of HPHV steam sterilizer (Bung processor ) respective SOP.
  • Record the PLC set parameters for the sterilization cycle during loaded chamber heat penetration study in the formats of Exhibit-E09.
  • Connect the probes to a calibrated data logger, which can scan and print the actual temperature observed at different locations with respect to time.
  • Place the probes and BI inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement.
  • When the sterilization cycle is complete, collect Strip chart/ printout of the sterilizer and attach to Exhibit-E09.
  • Download the data from data logger into the computer for data-analysis and printing. Attach the data printouts to Exhibit-E09.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. The observations shall be recorded in the formats of Exhibit-E09.
  • External temperature probe placement and justification for location of selection for Miscellaneous Items Load as per table given below
  • Probe  No. Location In The Chamber Justification For Location Selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    EP 2 Inside Pressure vessel 20 L To verify the temperature penetration in the innermost portion of these items.
    EP 3 Inside Pressure vessel 10 L
    EP 4 Inside Waste bin
    EP 5 Inside SS bucket
    EP 6 Inside Mopping pads pack
    EP 7 Inside Measuring cylinder
    EP 8 Inside silicone tubing
    EP 9 Inside cartridge filter
    EP 10 Housing for cartridge filters
    EP 11 Inside SS trays
    EP 12 Mop stick (At the head)
    EP 13 50 mm above the load, in the top rack aseptic side To verify the temperature distribution above the packs in the bottom rack.
    EP 14 50 mm above the load, in the middle rack aseptic side
    EP 15 50 mm above the load, in the middle rack non-aseptic side
    EP 16 50 mm above the load, in the bottom rack aseptic side

    (Note: The temperature probe shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels).

    • The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistency of temperatures.
    • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the Miscellaneous Items Load. The data shall be compiled from the three runs each load shall be reviewed and a report of the heat penetration study shall be prepared and recorded as per Exhibit-E09.

    External temperature probe placement and location for Miscellaneous Items Load:6

5.2.4.5 Heat Penetration Study – Non-RFS Rubber Stoppers

5.2.4.5.1 For Loading pattern and Set parameters refer SOP operation of HPHV steam sterilizer (Bung processor).

5.2.4.5.2 Procedure for the spiking of the bungs with endotoxin

  • The Endotoxin indicator vial containing 100, 000 EU / Vial shall be used for endotoxin spiking. The endotoxin vials shall be rehydrated with 1.0 ml of LRW to get 100,000 EU/ml.
  • After rehydration, endotoxin Indicator vial shall be shall be treated on a Vortex mixer for 2 minutes and then 1 min interval of 10 minute for one -half hour.
  • Dispense 0.1 ml of endotoxin into each rubber bung to be spiked with the Endotoxin.
  • Airs dry the spiked bungs by keeping under LAF for over night.
  • Each bung will now contain 10,000 EU.
  • Place spiked rubber bungs in the different carriage with proper identification mark in the Bung Processor.

5.2.4.5.3 Required Quantity of bungs for Endotoxin spiking

  • For minimum load of 20,000 rubber stoppers, 05 number of rubber bungs shall be spiked with 10,000 EU. 01 number shall be kept as positive control and 04 number of rubber bungs shall be distributed into four cassettes of the rotating carriage.
  • For maximum load of 80,000 rubber stoppers 17 number of rubber bungs shall be spiked with 10,000 EU. 01 number kept as positive control and 16 number of rubber bungs shall be distributed into sixteen cassettes of the rotating carriage. Each cassette shall have one spiked rubber bung.
  • This exercise shall be repeated for three consecutive loads.
  • The location of spiked rubber bungs shall be as per the defined pattern for minimum and maximum cycle.
  • Place the Endotoxin spiked bungs into the lad before start of the washing cycle.
  • Record the PLC set parameters for the washing and sterilization cycle during loaded chamber heat penetration study in the formats of Exhibit-E10 and Exhibit-E11 for Minimum load 20,000 and maximum load 80,000 stoppers respectively
  • After completion of the washing and siliconisation activity bye-pass the rotations of the rotary drum (to make it stationary during sterilization and drying period) of the bung processor to stop the cycle in order to place the temperature mapping in the inner most portion of the rubber stopper holding cassettes.
  • Connect the probes to a calibrated data logger, which can scan and print the actual temperature observed at different locations with respect to time.
  • Place the probes and BI inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement.
  • When the sterilization cycle is complete, collect Strip chart/ printout of the sterilizer and enclose to Exhibit-E11.
  • Collects the steam condensate as per respective SOP.
  • Aseptically collect the exposed biological indicators, spiked rubber bungs and send it to microbiology lab after wrapping in a sterile enclosure. The observations shall be recorded in the formats of Exhibit.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to recording Exhibits.
  • Recovery of Exposed Spiked Endotoxin Bungs:
    • After completion of the cycle take bungs from each location that was spiked with Endotoxin 10,000 EU.
    • Assuming that after washing and sterilization a three log reduction must have taken place, the total concentration on each bung will be 10 EU / bung.
    • Take processed rubber bungs into different depyrogenated glass tube and Add 5.0 ml of LRW into the glass tube with the help of a calibrated pipette.
    • Apply parafilm over the mouth of the glass tube.
    • Vortex tube for at least 15 min. The total concentration in glass tube will be 2.0 EU / ml. — (1:5).
    • Follow the dilutions as per the table for determining the content of Endotoxin in the Bung.
  • S.No. Endotoxin Concentration Total Dilution
    1 0.2 mL of 2 EU/mL     + 1.4 mL LRW 0.25 EU/mL 1:8
    2 1.0 mL of 0.25 EU/mL     + 1.0 mL LRW 0.125 EU/mL 1:16
    • Test the dilutions at S.No. 1 and 2 in duplicate using the LAL having confirmed labeled sensitivity equal to 0.125 EU/ml.
    • Analyze the sample as per the GTP- 206 for endotoxin testing.

     

    •      Perform the same exercise for each spiked and exposed bung and for the  

          other two loads.

    • Result shall be recorded in Exhibit-E10 for Non RFS rubber stoppers minimum 20,000 load, Exhibit-E11 for Non RFS rubber stoppers maximum 80,000 load.
    • Recovery of Unexposed Spiked Endotoxin Bungs
      • Reconstitute the Endotoxin indicator bung containing 10,000 EU/Bung with 5 ml of LRW in to the glass tube.
      • Vortex for a minimum of 15 min. and make further dilutions as shown below.
      • The total concentration in Bung will be 2000 EU / ml. ———— (1:5).
      • Vortex each dilution for at least 1 min. before making further dilution.
    S.No.  Endotoxin Concentration      Total Concentration Dilution
    1 0.1 mL of 2000 EU/mL + 1.9 mL of LRW 100 EU/mL 1:20
    2 0.1 mL of 100 EU/mL  + 9.9 mL of LRW 1 EU/mL 1:2000
    3 0.1 mL of 1 EU/mL  + 0.1 mL of LRW 0.5 EU/mL 1:4000
    4 0.1 mL of 0.5 EU/mL  + 0.1 mL of LRW 0.25 EU/mL 1:8000
    5 0.1 mL of 0.25 EU/mL  + 0.1 mL of LRW 0.125 EU/mL 1:16000
    6 0.1 mL of 0.125 EU/mL  + 0.1 mL of LRW 0.06 EU/mL 1:32000
    7 0.1 mL of 0.06 EU/mL  + 0.1 mL of LRW 0.03 EU/mL 1:64000
    • Test the dilutions at S.No. 4, 5 and 6 in duplicate using the LAL having confirmed labelled sensitivity equal to 0.125 EU/ml.
    • Analyze the sample as per the GTP -206 for endotoxin testing.
    • Perform the same exercise for the other two loads.

     

    • Temperature probe placement and justification for location of selection for non RFS stoppers load minimum 20,000, refer table below
    Probe  No. Location In The Chamber Justification For Location Selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    EP 2 Inside rubber stopper cassette – 1 To verify the temperature penetration in the innermost portion of the load.
    EP 3 Inside rubber stopper cassette – 3
    EP 4 Inside rubber stopper cassette – 5
    EP 5 Inside rubber stopper cassette – 7
    EP 6 Inside empty cassette – 2 To verify the temperature distribution in the empty cassettes.
    EP 7 Inside empty cassette – 4
    EP 8 Inside empty cassette – 6
    EP 9 Inside empty cassette – 8
    EP 10 Inside empty cassette – 9
    EP 11 Inside empty cassette – 10
    EP 12 Inside empty cassette – 11
    EP 13 Inside empty cassette – 12
    EP 14 Inside empty cassette – 13
    EP 15 Inside empty cassette – 14
    EP 16 Inside empty cassette – 15

    (Note: The temperature probe shall be placed in the pre determined locations with pre-identified probe numbers corresponding to the data logger channels).
       

    • Temperature probe placement, Endotoxin spiked bungs placement and BI placement for non RFS minimum stoppers load:7
      • External Temperature probe placement and justification for location of selection for non RFS stoppers maximum load:
      Probe No. Location in the chamber Justification for location selection
      EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
      EP 2 Inside rubber stopper cassette – 1 To verify the temperature penetration in the innermost portion of the load.
      EP 3 Inside rubber stopper cassette – 2
      EP 4 Inside rubber stopper cassette – 3
      EP 5 Inside rubber stopper cassette – 4
      EP 6 Inside rubber stopper cassette – 5
      EP 7 Inside rubber stopper cassette – 6
      EP 8 Inside rubber stopper cassette – 7
      EP 9 Inside rubber stopper cassette – 8
      EP 10 Inside rubber stopper cassette – 9
      EP 11 Inside rubber stopper cassette – 10
      EP 12 Inside rubber stopper cassette – 11
      EP 13 Inside rubber stopper cassette – 12
      EP 14 Inside rubber stopper cassette – 13
      EP 15 Inside rubber stopper cassette – 14
      EP 16 Inside rubber stopper cassette – 15

      (Note: The temperature probes shall be placed in the pre determined locations with pre-identified probes numbers corresponding to the data logger channels).
       

      • Temperature probe placement, Endotoxin spiked bungs placement and BI placement for non RFS Maximum Stoppers Load:8
        • The temperature data exhibited by external probes and in-built probes shall be compared to demonstrate consistently show the temperature variation between them
        • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the maximum and minimum non RFS stoppers load. The data compiled from the three runs each load shall be reviewed and a report of the heat penetration study shall be prepared and approved
        • The endotoxin spiked bungs demonstrate >3 log reduction

5.2.4.6 Heat Penetration Study for Liquid Nutrient Media

  • For Loading pattern and Set parameters refer SOP operation of HPHV steam sterilizer (Bung processor) respective SOP.
  • Record the PLC set parameters for the Liquid Nutrient media in the formats of Exhibit-E12.
  • Place the probes inside the load components, which are supposed to be most difficult points for steam penetration, these placements shall be uniform as given in the justification of temperature probe placement.
  • When the sterilization cycle is complete, collect Strip chart/ printout of the sterilizer and attach to Exhibit-E12.
  • Collects the steam condensate as per respective SOP.
  • Download the data from data logger into the computer for data-analysis and printing. Attach the data printouts to Exhibit-E12.
  • The observations shall be recorded in the formats of Exhibit-E12.
  • Compile the data generated during the qualification study for evaluation of the sterilization process efficacy for the Liquid Nutrient Media load. The data compiled from the three runs each load shall be reviewed and a report of the heat penetration study shall be prepared and approved
     

    • Temperature probe placement and justification for location of selection for Liquid Nutrient media. as per table given below:
    Probe  No. Location In The Chamber Justification For Location Selection
    EP 1 In the active Chamber Drain This is the location for the reference measurement point of the sterilizer, which controls the sterilization cycle. Hence important to compare the achieved temperature distribution results (strip chart/ print out of sterilizer) with the results from data logger.
    EP 2 Inside pressure vessel – 1 To verify the temperature penetration in the innermost portion of the load.
    EP 3 Inside pressure vessel – 2
    EP 4 Inside pressure vessel – 3
    EP 5 Inside pressure vessel – 4
    EP 6 Inside pressure vessel – 5 To verify the temperature distribution in the empty cassettes.
    EP 7 Inside pressure vessel – 6
    EP 8 50 mm above the load, in the top rack aseptic side
    EP 9 50 mm above the load, in the middle rack aseptic side
    EP 10 50 mm above the load, in the bottom  rack aseptic side
    EP 11 50 mm above the load, in the top rack non aseptic side
    EP 12 50 mm above the load, in the middle rack non-aseptic side
    EP 13 50 mm above the load, in the middle rack non-aseptic side
    EP 14 50 mm above the load, in the middle rack left side
    EP 15 50 mm above the load, in the top rack left side
    EP 16 50 mm above the load, in the top rack right side

    (Note: The temperature probe shall be placed in the pre determined locations with pre-identifed probe numbers corresponding to the data logger channels).
     

    • External Temperature probe placement for Liquid Nutrient media. Load:9

5.2.5 Vacuum Break Filter Integrity Testing

  • Objective of this test is to ensure that the sterilizing grade vacuum break filter installed in the aseptic side remains intact throughout the validation cycle.
  • Filter integrity test shall be carried out as per respective SOP and recorded in Exhibit-E13.

6.0 Acceptance Criteria:

The acceptance criteria for the tests parameters as defined above shall remains as defined below:

S. No. Test Parameters Acceptance Criteria
1 Verification of Calibration Should be Calibrated
2 Steam Quality Tests
3 Non-condensable gases Not More Than 3.5 %,
Superheat Not More Than 25°C.
Steam Dryness Value Not Less Than 0.90 (if metal loads are to be processed, the dryness value should not be less than 0.95).
4 Vacuum Leak Test Vacuum Leak Rate should be NMT 0.013 Bar / 10 minutes or 1.3 mbar/min.
5 Bowie –Dick Test There should be a uniform colour change throughout the indicator.No change, non-uniform change and/ or air entrapment (bubble) spot on the pattern indicates inadequate air removal from the sterilization chamber.
6 Empty Chamber Heat Distribution study The temperature indicated by the external and internal probes for the three cycles shall not be less than 121.00C and not more than 124.00C during the 30 minutes hold time.The temperature difference between the external probes and internal probes for each cycle during 30 minutes  hold time shall not be more than 2 0C.Any temperature probe in a location that attains a sterilization temperature of 121ºC after one minute with respect to the reference measurement point of the sterilizer (probe in the drain) is considered as cold spot.
7 Heat Penetration Studies The temperature exhibited by the external and internal probes for the three cycles shall not be less than 121.00C and not more than 124.00C during the 30 minutes hold time.The temperature difference between the external probes and in-built probes for each cycle during 30 min  hold time shall not be more than 2 0C.Biological indicators exposed to the steam sterilization cycle and incubated BI shall show no growth and positive controls should pass.Sterility Assurance Level when calculated using biological indicator D-value should demonstrate 10-6 (NLT 12 log reduction of spores).Specific Tests for endotoxin Spiked Bungs:Endotoxin spiked bungs if exposed for the washing cycle should give >3 log reduction in the level of Endotoxin.
8 Vacuum Break Filter Integrity Testing Water Intrusion Test value should be NMT 0.12 ml/ minute.

 

7.0 Validation Report:

The validation report shall consist of a summary document, which shall describe the activities carried out during the validation exercise. The validation report shall also detail the results of the validation exercise and the inferences drawn from it.

8.0 Approval of Validation Report:.

This validation report shall be reviewed and approved by Quality Assurance.

The following documents shall be maintained along with.

  • Approved validation protocol.
  • Summary reports of study.
  • Analysis and results of the runs.

9.0 Observed Deviation:

Record the Discrepancy and Corrective Actions during the study in the Exhibit-E14.

 10.0 List of Exhibit / Annexure:

10.1    List of Exhibits:

 Exhibit No. Annexure Title No. of Pages
E01 Vacuum Leak Test
E02 Bowie Dick Test
E03 Heat Distribution Empty Chamber
E04 Heat Penetration Study Garment Load-Minimum
E05 Heat Penetration Study Garment Load-Maximum
E06 Heat Penetration Study – Rubber Stoppers in RFS Bags-20 mm
E07 Heat Penetration Study – Rubber Stoppers in RFS Bags-32mm
E08 Heat Penetration Study-Sterile Powder Filling Machine Parts
E09 Heat Penetration Study – Miscellaneous Item Load
E10 Heat Penetration Study – Non RFS rubber stoppers minimum 20,000 load
E11 Heat Penetration Study – Non RFS rubber stoppers Maximum 40,000 load
E12 Heat Penetration Study – Liquid Nutrient Media
E13 Vacuum Break Filter Integrity Testing
E14 Discrepancy And Corrective Action Report

 

10.2     List of Attachments:

S. No. Title
   
   
   
   
   
   
   

 

11.0 Reference Documents:

  • Validation Master Plan.
  • SOP on operation of HPHV steam sterilizer
  • SOP on operation of Vacuum Leak test in bung processor
  • SOP on Bowie Dick test
  • SOP on Sterilization of Garments
  • SOP for washing, siliconisation and sterilization of Rubber stoppers

12.0 Periodic Validation Schedule:

Schedule Validation Frequency
Study Periodic -validation Routine
Calibration of   all instruments e.g., Pressure Gauge, Compound   Gauges, Timer, Data logger, Probes. Six month Every Month for Pressure Gauge, Compound   Gauges.
Vacuum Hold Test Six month (Pre and Post validation) Every Day one cycle
Steam quality test Six month Not required
Bowie Dick Test Six month Once in a week one cycle.
Empty-chamber heat distribution studies (3 cycles) Six month Not required
Loaded   chamber heat distribution and heat penetration studies (3 Cycles/Load) Six month Not required

12.1 Revalidation:

Revalidation shall be carried out in case of Major maintenance of critical parts; Change of cycle program, Inclusion of new load.

For unscheduled revalidation, Vacuum leak rate test with temperature mapping sensors, Empty chamber and loaded chamber revalidation shall be carried out. The pattern of the study shall be based on case-to-case basis and shall be documented.

13.0 Revision History:

  • Reference of for steam quality test, Bowie Dick test and Vacuum leak test incorporated in protocol
  • Preventive maintenance and calibration status mentioned in protocol.
  • Diagrammatically representation of the probes and Biological Indicators incorporated in the protocol.
  • Periodic validation and revalidation criteria and schedule added.
  • Revalidation schedule changed to once a year.
    • SOP on monitoring of purified water, water for injection and pure steam condensate
    • GTP For endotoxin testing.
    • SOP for Steam Quality Testing

END OF THE DOCUMENT

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