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Performance Qualification Protocol. P.H.V. Steam Sterilizer

  • OBJECTIVE
  • The objective of this Validation Protocol is to obtain a high degree of assurance that the steam sterilization process is capable of sterilizing the specified loads using High Pressure and High Vacuum Steam Sterilizer (Bung Processor).
  • To establish a documented evidence that the Steam sterilizer, as commissioned, will produce acceptable goods when operated in accordance with the process specification.
  • The sterilization process consistently provides a specified degree of sterility assurance level for each type of load selected for sterilization.
  • Also the utilities supplied will meet the predefined specifications and capable of supporting the sterilization process consistently.
  • Different test procedures to be conducted as described in the preceding sections along with the acceptance criteria to meet for each type of test will qualify the equipment.
  • SCOPE
  • This protocol is applicable to P.H.V. Steam Steriliser 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.
  • 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
ºC Degree Celsius
Temp. Temperature
Min. Minimum
Max. Maximum
  • Fο value: A quantity, measured in minutes, used to determine the efficacy of an operating cycle and equivalent to a continuous period at a temperature of 121°C.
  • D-value: Decimal reduction value (for biological indicators). The time in minutes required to secure inactivation of 90% of the test organisms under stated exposure conditions.
  • EQUIPMENT DESCRIPTION
  • H.P.H.V. Steam sterilizer is installed in vial washing room.
  • Purpose Of The Equipment
  • Purpose of this H.P.H.V. Steam sterilizer is for sterilization and drying of the garments, rubber stopper holding canisters, filling machine parts, filtration accessories and previously washed and siliconised rubber stoppers.
  • Basic Mode Of Operation
  • The main action of H.P.H.V. Steam sterilizer on all the materials placed within the chamber is that, the steam penetrates into the microorganisms present on and within the material thereby transferring the latent heat, which in turn will coagulate the proteins, imparting a lethal effect on the microorganism. This will lead to the destruction of living organisms.
  • The H.P.H.V. Steam sterilizer is made-up of SS sheet, which is welded with a ‘U’ profile SS jacket.
  • The sterilization chamber is provided with two SS sliding doors reinforced with MS support structure. The door is operated with the help of pneumatic cylinders. When both the doors reach the end position the gaskets are pushed out automatically with the help of compressed air ejectors. When the gaskets retract the sterilizer chamber, door slides automatically.
  • The door sealing is made effective with the help of tubular silicon gasket to ensure proper sealing.
  • These gaskets are activated with compressed air and retracted with the help of vacuum.
  • The sterilization chamber is insulated with resin-bonded glass wool, which helps in reducing the heat loss to the environment and ensures uniform distribution of temperature inside the chamber. The insulation is covered with SS cover plate.
  • To ensure leak tight partition between the sterile area and the loading side a SS flush panel is provided on the partition wall and the outer cover of the Bung processor.
  • All the joints, crevices are filled with silicon sealant to prevent Leakages.
  • The autoclave holds the vacuum with a rate of NMT 13 mbar per 10 minutes.
  • All the automatic valves are closed when autoclave chamber is at the atmospheric pressure. During the vacuum cycle except the vacuum valve all other valves are closed.
  • The sterilizing grade filters are suitable for integrity testing and steam sterilization.
  • Both the horizontal sliding doors have an interlocking system in which the door doesn’t open simultaneously. The doors doesn’t open in any case during sterilization.
  • The Bung processor has the ability to withstand maximum of 3.0 kg / cm2 pressure. The system is having an emergency stop so as to stop all the physical movement and operation of the machine immediately, which is located along with the control panel.
  • H.P.H.V. Steam sterilizer, option between four pre-programmed cycles is available.
  • Vacuum leak test
  • Bowie-dick test
  • Standard sterilization cycle
  • P.H.V. sterilization cycle / H.P.H.V. Sterilization cycle for bungs
  • Process Description
  • The sterilizer is supplied with pure (clean) steam for the chamber and plant steam to the jacket.
  • The Sterilization cycle can be automatically controlled by the PLC, which can be programmed and protected with three different levels of passwords.
  • The 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.
  • HPHV cycle.
  • High Pressure High Vacuum sterilization cycle with vacuum pulses.
  • All the above cycles are PLC controlled and runs automatically when selected.
  • Operating cycle for a porous load
  • The selected operating cycle of a porous load sterilizer normally 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.
  • Area Description
  • The HPHV steam sterilizer is located in the washing & sterilization room of the sterile powder injectable filling section with restricted access.
  • As per the specifications for cleanliness the washing area is designed as class D & sterilization room is designed Grade – C.
  • 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 washing & sterilization room for loading and the other door opens into the aseptic area for unloading.
  • The control panel is located in the washing & sterilization room; a modular room physically separates its major components and utility lines from the washing & sterilization room environment.
  • The other supporting systems such as, water-ring vacuum pump is located near the sterilizer and the pure steam generator is located in the Utility building, in the area dedicated for water system.
  • 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.
  • The following table shows the tests to be done on the steam sterilizer.
1 Steam qualification tests

·   Non-condensable gases test

·   Steam super heat test

·   Steam dryness test

3 runs of each test
2 Vacuum Leak test 3 runs
3 Empty Chamber Heat Distribution study ·With temperature mapping probes at different locations of the sterilizer chamber.

Bio-Challenge studies shall use Bacillus stearothemophilus spore strips (containing 106 or more spores per strip) during the heat distribution studies.

4 Bowie –Dick Test 3 Trials on 3 different days
6 Loaded Chamber heat penetration studies

(The following are the fixed standard load patterns selected for qualification of the sterilizer and performed for 3 runs)

·  Aseptic area Garments

·  Filling machine parts

·  Filtration accessories

·  Rubber stopper holding canisters

·  Rubber stoppers

·  Media vessels with WFI water

 

·       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 Bacillus stearothemophilus spore strips (containing 106 or more spores per strip) during the heat penetration studies.
Estimation of the FO value achieved during the sterilization hold period at each temperature-mapping probe.
  • Steam Quality Tests
  • Steam Non-Condensable Gas Test
  • Tools required: Steam quality Testing kit
  • Test procedure:
  • The apparatus is shown and described in Figure NO.1 (all sizes are nominal).
  • Connect the needle valve to the steam service pipe as shown in Figure NO.1
  • Assemble the apparatus so that condensate will drain freely from the long rubber tube into the sampling pipe. If the tube is too short, copper or stainless steel tubing may also be used.
  • Fill the container with cold water until it overflows. Fill the burette and funnel with cold water, invert them and place them in the container. Draw out any air that has collected in the burette.
  • With the steam sampling pipe out of the container, open the needle valve and allow steam to purge the air form the pipe.
  • Place the pipe in the container, locate the end within the funnel, and add more cold water until it flows through the overflow pipe.
  • Place the empty measuring cylinder under the container overflow.
  • Adjust the needle valve to allow a continuous sample of steam into the funnel sufficient to cause a small amount of “Steam Hammer” to be heard. Ensure that all the steam is discharged into the funnel and does not bubble out into the container. Note the setting of the needle valve. Close the valve.
  • Ensure that the container is topped up with cold water and that the measuring cylinder is empty. Draw out any air present in the burette.
  • Ensure that the sterilizer chamber is empty except for the usual chamber furniture. Select and start the operating cycle.
  • When the steam supply to the chamber first opens, open the needle valve to the previously noted setting, allowing a continuous sample of steam into the funnel sufficient to cause a small amount of steam hammer to be heard.
  • Allow the steam sample to condense in the funnel. Any non-condensable gases will rise to the top of the burette. Overspill formed by the condensate and the water displaced by the gases will collect in the measuring cylinder.
  • When the temperature of the water in the container reaches 70-75°C close the needle valve. Note the volume of gas collected in the burette (Vb) and the volume of water collected in the measuring cylinder (Vc).
  • Calculate the fraction of non-condensable gases as a percentage as follows.
  • This test is used to demonstrate that the level of non-condensable gases in the steam will not prevent the attainment of sterilization conditions in any part of the load. The method described should be regarded not as measuring the exact level of non-condensable gas, but a method by which the provision of acceptable steam quality can be demonstrated.
  • Fraction of non-condensable gases = 100 x (V b/V c).
  • The test should be done two more times to check consistency. If the results of the three tests differ significantly, then the cause should be investigated before proceeding further.
  • Record the observations & results in the annexure-1.
  • Acceptance Criteria: The measured non-condensable gases in the pure steam should not cross 3.5%
  • Steam Super Heat Test
  • Tools required: Testing kit
  • Test procedure:
  • This test should normally follow a satisfactory test for non-condensable gases.
  • This test, and the subsequent dryness value test, requires a pitot tube as shown in figure No.2 . The rest of the apparatus is shown and described in figure No.3. All sizes are nominal.
  • Fit the Pitot tube concentrically within the steam service pipe as shown in figure No.3.
  • Fit the sensor entry gland to the steam service pipe. Insert one of the sensors through the gland and position the axis of the pipe.
  • Insert the second sensor through the gland in the expansion tube and position it on the axis of the pipe. Wrap lagging around the expansion tube. Push the tube on to the pitot.
  • Ensure that the sterilizer chamber is empty except for the usual chamber furniture.
  • Select and start the operating cycle.
  • From the measured temperatures, note the temperature in the steam service pipe (for use in the dryness test) and in the expansion tube (Te) when the steam supply to the chamber first opens. Calculate the superheat in oC from the following equation:
  • Superheat  = Te – To
  • Where:
  • To  is the boiling point of water at local atmospheric pressure.
  • Record the observations & results in the annexure-1.
  • Acceptance criteria: The test should be considered satisfactory if the superheat measured in the expansion tube does not exceed 25 ºC
  • Steam Dryness Test
  • Tools required: Testing kit, balance
  • Test procedure:
  • The test is conveniently carried out immediately after the superheat test.
  • This test requires a pitot tube as shown in Figure No.2. The apparatus is shown and described in figure No.3. All sizes are nominal. A laboratory balance is also required, capable of weighing a load up to 2 kg with an accuracy of 0.1g or better.
  • If it is not already fitted, fit the Pitot tube concentrically with in the steam service pipe as shown in figure No.3.
  • If it is not already fitted, fit the sensor entry gland to the steam service pipe. Insert a temperature sensor through the gland and position it on the axis of the pipe.
  • Connect the rubber tube to the longer of the pipes in the stopper, place the stopper in the neck of the vacuum flask, weigh the whole assembly and note the mass (M1).
  • Remove the stopper and tube assembly and pour 650 +_ 50 ml of cold water (below 27ºC) in to the flask. Replace the stopper and tube assembly, weigh the flask and record the mass (M2).
  • Support the flask close to the pitot, and ensure that the rubber tube and flask are protected from excess heat and draughts, don’t connect it to the Pitot tube yet.
  • Introduce the second temperature sensor through the shorter of the two pipes in the stopper and into the water in the flask. Note the temperature of the water in the flask (To).
  • Ensure that the sterilizer chamber is empty except for the usual chamber furniture. Select and start the operating cycle.
  • When the steam supply to the chamber first opens, connect the rubber tube to the pitot discharge and wrap lagging around it. Arrange the rubber tube to permit condensate to drain freely into the flask. Not the temperature in the steam service pipe (TS).
  • When the temperature of the water in the flask is approximately 80°C, disconnect the rubber tube from the pitot, agitate the flask so that the contents are thoroughly mixed, and note the temperature of the water (T1).
  • Weigh the flask and stopper assembly and note the mass (M3).
  • The initial mass of water in the flask is given by Mw = M2 – M1
  • The mass of condensate collected is given by MC = M3 – M2.
  • Calculate the dryness value of the steam from the following equation:
  • D= (T1-T0) (4.18MW + 0.24) / LMC  – 4.18 (TS-T1) / L

Where:

T0 =  Initial temperature of the water in the flask (oC);

T1 = Final temperature of the water and condensate in the flask (ºC);

TS = Average temperature of the steam delivered to the sterilizer (ºC);

MW  = Initial mass of water in the flask (Kg);

MC = Mass of condensate collected (Kg);

L = latent heat of dry saturated steam at temperature TS (kJ Kg-1).

  • Record the observations in the annexure – 1.
  • Therefore, the minimum F0 value required for more than 6 log reduction of the Bacillus sterothermophilus indicator should not be less than 20.95 minutes.
  • Fο = 20.95 min.
  • Fο = 1.7 x (log 2.1 x 106 –log 10-6)
  • Min., spore population (A) = 2.1 x 106 are used for bio-challenge study of a sterilization cycle, which is designed to achieve 12-log reduction, i.e. PNSU 10-6. Biological F0 value for this indicator can be calculated by equation (b) and the results are
  • Example – Biological indicator strips supplied by … Co., Lot # … Expiry … D121 =1.7
  • B = Desired level of non-sterility
  • A = Biological indicator concentration or spore population
  • Where, D121 = D value of the biological indicator at 121ºC
  • Fο = D121 (log A – log B) —-(a)
  • Calculate the required Fο value for biological indicator exposed during the sterilization as per the formula given below.
  • Required F0 Value Calculation For The Biological Indicator Used
  • F0 VALUE CALCULATION FOR THE BIOLOGICAL INDICATOR USED
  • An unsatisfactory test result indicates that the machine should not be used until the fault has been rectified. (It is important to realise that if a sterilizer fails to pass the Bowie-Dick test it cannot be made safe simply by increasing the holding time until a uniform colour change is produced. A failed sterilizer is in urgent need of skilled attention and f
  • The indicator paper should be marked with the result and kept for reference for at least three months. (The chemical reaction continues during this time and the paper may be discarded when the indicator becomes unreadable.)
  • It is important to compare the colour of the indicator at the corners of the paper with that at the centre so that any difference can be clearly seen. If there is any discernible difference the test should be recorded as failed, and the paper marked accordingly. A large area of unchanged indicator points to a gross failure.
  • No change, non-uniform change and/ or air entrapment (bubble) spot on the pattern indicates inadequate air removal from the sterilization chamber.
  • The automatic controller indicates that a Bowie-Dick test cycle has just been completed.
  • There is a uniform change throughout the indicator;
  • The test should be considered satisfactory if the following requirements are met:
  • Acceptance Criteria
  • The printout/ strip chart taken during the Bowie-Dick test cycle & the Bowie-Dick test indicator should be preserved along with annexure-1.
  • When the cycle is complete, remove the indicator paper from the test pack.
  • Start the operating cycle.
  • Ensure that the holding time will not be longer than that specified above. (If this time is exceeded, the indicator may change in such a way as to make it difficult to detect the variations that would indicate a fail condition).
  • Select the Bowie-Dick on the PLC, to operate the steam sterilizer automatically.
  • Place the test pack in the chamber with the bottom of the pack supported 100-200 mm above the centre of the chamber base.
  • Remove the wrapping from a standard test pack and place the indicator paper in the sheet located nearest to the centre of the pack. Reassemble and secure the pack and replace the wrapping.
  • The Bowie-Dick test is normally preceded by a warm-up cycle. This cycle is necessary because the effectiveness of air removal may depend on all parts of the sterilizer being at working temperature. A satisfactory sterilizer may give a fail result if this is not done.
  • Operate the Steam sterilized as per the SOP set the PLC parameters.
  • Test procedure:
  • Tools required: The Bowie-Dick test uses a Class B chemical indicator contained within a standard test pack.
  • Acceptance Criteria
  • The absolute pressure (P2) at the start of the 10-minute period is less than 70 mbar.
  • The vacuum leak rate does not exceed 1.3mbar/min or the rate of vacuum drop at the end of 10 minutes holding time should not be more than 13 mbar.
  • BOWIE – DICK TEST FOR STEAM PENETRATION
  • Sterilization is achieved by the rapid and even penetration of steam into all parts of the load and the maintenance of these conditions for the specified holding time. To ensure this, it is essential to remove air from the chamber and load, and to provide a steam supply, which contains a minimal volume of non-condensable gases. Any residual air and non-condensable gases will become concentrated as a bubble in the load and inhibit steam penetration.
  • The Bowie-Dick test shows whether or not steam penetration of the test pack is even and rapid, and thus by implication that air or other non-condensable gases are not present. It does not confirm that the sterilization conditions in the load have been achieved.
  • Record the observations & results in the annexure-1.
  • Acceptance Criteria
  • The test should be considered satisfactory if the following requirements are met.
  • The dryness value is not less than 0.90 (if metal loads are to be processed, the dryness value should not be less than 0.95);
  • Through out the operating cycle, the temperature measured in the steam service pipe is with in 3ºC of that measured during the superheat test.
  • VACUUM LEAK TEST
  • Tools required: Calibrated Data logger
  • Test procedure:
  • Operate the Steam sterilized as per the SOP.
  • Measuring the change of vacuum in the chamber when all valves leading to it have been closed and the vacuum source isolated performs the test.
  • Operate the steam sterilizer empty. Allow the sterilization chamber temperature to stabilize.
  • When the chamber temperature is stabilized, the vacuum pump starts.
  • Achieve chamber pressure (on gauge) equivalent to ≤≥ 100-mbar of absolute pressure. Observe channel No.6 of the multi point recorder for vacuum).
  • When the pressure in the chamber drops below 50 mbar absolute, close all the valves connected to the chamber and stop the vacuum pump. Note the time and the absolute Pressure P1 (achieved in the sterilizer chamber).
  • Wait for 5 minutes (± 10 s) to stabilize the chamber pressure (This time allowance is provided to allow evaporation of condensate in the chamber) and record the Pressure again (P2).
  • Wait for further 10 minutes (± 10 s), and then note the pressure for a third time (P3).
  • Calculate rate of Vacuum drop in the sterilizer chamber by using the equation given below and record the results in Attachment.
  • Calculate the vacuum leak rate for minute period from the equation:
  • Vacuum leak rate = (P3-P2) / mbarmin-1
  • Where, P2 = Pressure (vacuum) observed after 5 minutes of achieving the desire level of pressure.
  • P3 = Pressure (vacuum) observed after 10 minutes of achieving  pressure P2
  • Record the observations and results in annexure-1.
  • The printout/ strip chart taken during the test should be enclosed as attachement.
  • Z = the change in the heat resistance of Bacillus Stearothermophilus spores as temperature is changed (10ºC).
  • T  = the observed temperature at that particular time (as per the actual temperatures recorded)
  • Where, Dt = the time interval between successive temperature  measurements (1 min.).
  • Fο = Dt ∑ (Sum of lethality factors)
  • Fο = Dt ∑ 10(T-121)/Z         ……………  (b)
  • The calculations are carried out automatically by using the following equation (b) and the lethality factor computed is given in the cumulative lethality chart (printout) and Fo chart (bar graph printout).
  • The actual observations obtained during the Heat distribution & Penetration studies at different locations were obtained in the temperature data (printout) and the observed temperatures were subjected for calculation of Fο values at that particular location.
  • Fο Value Calculation For The Biological Indicator Used after Heat Distribution And Penetration Study.
  • Acceptance Criteria: The Calculated minimum Fο value by equation (a) (i.e. from the cumulative lethality chart) should be more than the biological Fο value for the biological indicator exposed for the bio-challenge study.
  • HEAT DISTRIBUTION EMPTY CHAMBER
  • Tools required: Calibrated data logger.
  • Test procedure:
  • Pass predifined no. of 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 as per annexure-2 so that probes do not touch any metallic surface, also place biological indicators along with each temperature-mapping probe in the sterilizer chamber.
  • Connect the probes to a suitable data logger, which can scan and print the actual temperature observed at different locations with respect to time.
  • Select HPHV sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per 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 sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the Fο value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab.
  • If the temperature 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.
  • Record all the observations in the annexure-1.
  • Acceptance criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 2 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • HEAT DISTRIBUTION AND PENETRATION STUDIES
  • Heat Penetration Study Garment Load
  • Load material: as per the table below.
12 pairs of aseptic area gowning Each one number of trunk, pair of booties and headgear in one cloth bag. Four such bags are placed side by side as one layer in the trolley and three such layers are loaded amounting to 12 pairs.
Three numbers of glove packs Each containing 8 pairs of powder free latex gloves.
12 numbers of lint free mopping dusters Wrapped using parachment paper and arranged side by side
  • Test procedure:
  • Pass define no. of 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select HPHV sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the F0 value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • Heat Penetration Study-Filling Machine Parts
  • Load material: Following parts are included in the load
  • Powder hopper – 1 number
  • Rubber stopper hopper – 1 number
  • Rubber stopper hopper chute – 1 number
  • Powder wheel with pistons – 10 numbers
  • 1 number of 0.2-micron Catridge filter with housing for N2/ CO2 filtration
  • Nos. of vent filters of 0.2-micron porosity for nitrogen filtration at dosing stations
  • Suitably wrapped silicone tubing & Forceps
  • Test procedure:
  • Pass pre-define no. of 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select HPHV sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the F0 value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • Heat Penetration Study – Filtration Accessories
  • Tools required: Load material as follows.
  • 293 mm diameter membrane holder previously washed with WFI and fitted with 0.2-micron N66 membrane filter and 2 micron pre-filter with suitable silicone tubing for disinfectant filtration.
  • 25-liter pressure vessel previously washed with WFI for storage of sterile filtered disinfectants.
  • Silicon tubing required for water filtration and clamps required for fixation of the membrane holder and tanks assembly.
  • Test procedure:
  • Pass pre-defined no. of 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select HPHV sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the Fο value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • Heat Penetration – Rubber Stoppers
  • Tools required: Load material as follows.
  • Nos. of previously washed and siliconised 20 mm stoppers loaded in 4 nos. of SS perforated cassettes ( Nos. approximately in each cassette amounting to nos.) These cassettes are loaded into the rotating carriage of the bung processor and the carriage is loaded into the chamber of the bung processor.
  • Test procedure:
  • Pass pre-defined no. of 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select HPHV sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the Fο value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • Heat Penetration Study – Rubber Stopper Holding Canisters
  • Load material: Nos. of rubber stopper holding canisters (without perforations and with lids) previously washed with WFI, used for holding washed-siliconised-sterilized-dried, rubber stoppers. Arranged 4 nos. in one row and 2 such rows are placed in the sterilizer chamber amounting to 8 nos. of canisters.
  • Test procedure:
  • Pass pre-defined no. of 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select HPHV sterilization cycle on control panel and set the parameters..
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the Fο value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required F0 value for the biological indicator.
  • Heat Penetration Study – Media vessels with WFI water
  • Load material: nos. of pressure vessels (used for media) contains WFI water around 17 – 18 liters placed in the sterilizer chamber.
  • Test procedure:
  • Pass  no. 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 inside the load components, which are supposed to be most difficult points for steam penetration, these placement shall be uniform and as per the annexure-2.
  • 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.
  • Select sterilization cycle on control panel and set the parameters.
  • Operate the steam sterilizer as per SOP and also start the data logger to record actual temperatures with in the sterilization chamber with respect to time. All the times set on PLC, Strip chart recorder, Printout and data logger must correspond to each other.
  • When the sterilization cycle is complete
  • Collect Strip chart from the Strip chart Recorder of the sterilizer and enclose to annexure-1 as attachment.
  • Download the data from data logger into the computer for data-analysis and printing. Enclose the data printouts to annexure-1 as attachment. Review and calculate the F0 value for each temperature probe.
  • Aseptically collect the exposed biological indicators and send the indicators after wrapping in a sterile enclosure to microbiology lab. Record the observations in the annexure-1.
  • If the results obtained from the heat penetration study are satisfactory, perform (repeat) two more times on each load to check for reproducibility and to establish permitted tolerances as described in the acceptance criteria.
  • Record all the observations in the annexure-1.
  • Acceptance Criteria
  • The measured temperatures in the sterilizer chamber should be uniform.
  • The temperature in the Sterilisation hold time should be within the sterilization temperature band of 121ºC to 124ºC.
  • Lag period should be NMT 5 minutes.
  • Biological indicators exposed to the steam sterilization cycle When aseptically collected and incubated should show no growth.
  • The calculated minimum Fο value should be more than the required Fο value for the biological indicator.
  • 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.

 

  • Figure-1

34

  • Figure-2

35

  • Figure-3

36

 

  • Forms and Records (Annexures)
  • Annexure – 1 (Recording Of Observations For Performance Qualification)
  • Annexure – 2 (Load pattern & Justification)
  • Distribution
  •  Quality Assurance, Production, Quality Control
  • History
    Date Revision Number Reason for Revision
    00 New SOP

 

Annexure – 1 (Recording Of Observations For Performance Qualification)

Details Of Operational Qualification Report
Report No.   Date Of Authorisation  

 

  Identification Of The Executors
S. No. Name Designation & Department  Sign & Date Training Details
       
       

Comments:

Reviewed by:                                                                                          Date:

  • Steam Quality Tests
  •  Steam Non-Condensable Gas Test
  • When the temperature of the water in the container reaches 70-75°C close the needle valve. Note the volume of gas collected in the burette (Vb) and the volume of water collected in the measuring cylinder (Vc).

Calculate the fraction of non-condensable gases as a percentage as follows.

  • Fraction of non-condensable gases = 100 x (V b/V c).
  • The test should be considered satisfactory if the fraction of non-condensable gases does not exceed   3.5 %.
Checks Observations
Run – 1 Run – 2 Run – 3
The volume of gas collected in the burette (Vb)      
The volume of water collected in the measuring cylinder (Vc)      
Fraction of non-condensable gases as percentage. (100 x (V b/V c))      
Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Comments:
  • Reviewed by:                                                                                          Date:
  • Super Heat Test
  • From the measured temperatures, note the temperature in the steam service pipe (for use in the dryness test) and in the expansion tube (Te) when the steam supply to the chamber first opens. Calculate the superheat in ºC from the following equation:
  • Superheat  = Te – To
  • Where:To  is the boiling point of water at local atmospheric pressure.
  • Acceptance criteria: The test should be considered satisfactory if the superheat measured in the expansion tube does not exceed 25ºC.
Checks Observations
Run – 1 Run – 2 Run – 3
Temperature in the expansion tube (Te)      
The boiling point of water at local atmospheric pressure. 100 oC 100 oC 100 oC
Superheat  = Te – To      
Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Comments:
  • Reviewed by:                                                                                          Date:

 

  • Steam Dryness Test
  • Dryness value of the steam from the following equation:
  • D = (T1-T0) (4.18MW + 0.24) / LMC  – 4.18 (TS-T1) / L
  • Where:
  • T0 = Initial temperature of the water in the flask (oC);
  • T1 =  Final temperature of the water and condensate in the flask (ºC);
  • TS = Average temperature of the steam delivered to the sterilizer (ºC);
  • MW = Initial mass of water in the flask (Kg);
  • MC = Mass of condensate collected (Kg);
  • L = latent heat of dry saturated steam at temperature TS (kJ Kg-1).
Checks Observations
Run – 1 Run – 2 Run – 3
Mass of Dryness test assembly (M1)      
Mass of flask with cold water, stopper & tube assembly (M2) in kg    
Temperature of the water in the flask before testing (T0)      
Temperature in the steam service pipe (TS)      
Temperature of the water in the flask after testing (T1)      
Mass of flask stopper & tube assembly after test (M3) in kg      
Initial mass of water in the flask is

MW = M2-M1 in kg

     
The mass of condensate collected is given by Mc= M3 – M2. in kg      
Dryness value of the steam as per the above formula      
Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Comments:
  • Reviewed by:                                                                                          Date:

 

 

Vacuum Leak Test

Checks Readings from the Strip chart
Run – 1 Run – 2 Run – 3
Initial

(P1)

After 5 min (P2) After 10 min (P3) Initial

(P1)

After 5 min (P2) After 10 min (P3) Initial

(P1)

After 5 min (P2) After 10 min (P3)
Date of Test
Sterilization Chamber Pressure
Sterilization Chamber Temperature
Done by

Sign & Date

Checked by

Sign & Date

  • Refer attachment (printouts) No.:________
  • Calculations:
  • Vacuum leak rate = (P3-P2) / mbarmin-1
  • Result: The vacuum leak rate in the sterilization chamber is within / not within the                                 acceptance limit (i.e., .1.3mbar/ min).

 

  • Comments:
  • Reviewed by:                                                                                          Date:
  • Bowie – Dick Test For Steam Penetration
  •  Details of Bowie-Dick test pack
  • Company Name …………………………………………………………………
  • Cat.No……………………….,Expiry……. …………….…,Lot No……. ……………….……
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Whether the test pack placed in the chamber with the bottom of the pack supported 100-200 mm above the centre of the chamber base.      
Steam steriler operated as per the PLC set parameters    
Strip chart print out shows the hold time as per the procedure mentioned      
Colour of the indicator is changed uniformly.      
Done by

Sign & Date

     
Checked by

Sign & Date

     
  •  Refer attachment (printouts) No.:________
  • Comments:
  •  Reviewed by:                                                                                          Date:

 

Fο Value Calculation For The Biological Indicator Used

  • Required Fο Value Calculation For The Biological Indicator Used
Biological Indicator Lot No.  
Expiry date  
D121 value  
Spore population  
Equation for required Fo value to get PNSU of 10-6

Or 12 log reduction in the biological indicator

Fο =D121 (logA-logB)
Calculation Fο =  _____(…………..…… –  …………………)
FO value required to get PNSU of 10-6 …………….Minutes
FO value required to get PNSU of 10-12 …………….Minutes
Done by

Sign & Date

Checked by

Sign & Date

  • Comments:
  • Reviewed by:                                                                                          Date:

 

 

Heat Distribution Empty Chamber

Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 2 minutes)

     
Fο value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum Fο values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

 

Biological Indicator Incubation DetailsHeat Distribution Empty Chamber
Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Distribution Empty Chamber

Run No. 2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Distribution Empty Chamber

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

 Heat Distribution And Penetration Studies
  • Heat Penetration Study Garment Load
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum F0 values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration – Study Garment Load

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration – Study Garment Load

Run No. 2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration – Study Garment Load

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

  • Heat Penetration Study – Filling Machine Parts 
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum Fο values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration Study-Filling Machine Parts

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study-Filling Machine Parts

Run No. 2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study-Filling Machine Parts

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

  • Heat Penetration Study – Filtration Accessories
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum F0 values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration Study-Filtration Accessories

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study-Filtration Accessories


Run No.
2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study-Filtration Accessories

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

  • Heat Penetration Study – Rubber Stoppers
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum Fο values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration Study – Rubber Stoppers

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study – Rubber Stoppers


Run No.
2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study – Rubber Stoppers

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

  • Heat Penetration Study – Rubber Stopper Holding Canisters
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum F0 values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration Study- Rubber Stopper Holding Canisters

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study- Rubber Stopper Holding Canisters

Run No. 2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2  

 

3  

 

4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study- Rubber Stopper Holding Canisters

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

  • Heat Penetration Study – Media Vessels With WFI Water
Checks Observations

Yes/No

Run – 1 Run – 2 Run – 3
Date of test      
The measured temperature in the sterilizer chamber is found uniform.      
The temperature in the Sterilisation hold time within the sterilization temperature band of 121 to 124ºC.    
Observed Lag period

(Should be NMT 5 minutes)

     
F0 value for the biological indicator of each location is complies with the requirement.      
Write the minimum & maximum F0 values. Min._____

Max._____

Min._____

Max._____

Min._____

Max._____

Done by

Sign & Date

     
Checked by

Sign & Date

     
  • Refer Attachment No.: _________
  • Comments:
  • Reviewed by:                                                                                          Date:

 

Biological Indicator Incubation Details

Heat Penetration Study- Media Vessels With WFI Water

Run No. 1 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study- Media Vessels With WFI Water

Run No. 2 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Biological Indicator Incubation Details

Heat Penetration Study- Media Vessels With WFI Water

Run No. 3 Date of Testing  
Lot No. of Media Date of Report
Details of Observation of Incubation
Biological Indicator

No.

Days
1 2 3 4 5 6 7 8 9 10 11 12 13 14
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
+ Ve Control  

 

–  Ve Control  

 

Observed by  

 

  • Note: ‘+ Ve’ Indicates growth; ‘-ve’ Indicates No growth
  • Remarks: No growth / growth observed in the inoculated tubes (s) indicates that all spores are killed / not killed on exposure of the subjected sterilization cycle.

 

Discrepancy And Corrective Action Report
Reference :

Discrepancy :

Corrective Action :

Satisfactorily completed? [Yes / No]:

Checked by / date:                                                  Reviewed and approved by / date:

Reference :

Discrepancy :

Corrective Action :

Satisfactorily completed? [Yes / No]:

Checked by / date:                                                  Reviewed and approved by / date:

Reference :

Discrepancy :

 

Corrective Action :

Satisfactorily completed? [Yes / No]:

Checked by / date:                                                  Reviewed and approved by / date:

Annexure – 2 (Load pattern & Justification)

  • Temperature Sensor Placement In The Empty Chamber And Justification For The Selection Of Location
Sensor No. Location In The Chamber Justification For Location Selection
1 In the active Chamber Drain (Near RTD  -01) 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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Non-sterile door middle  

Any conduction of heat through the door, which may cause temperature drop at that particular point.

7 Non-sterile door left corner bottom
8 Non-sterile door right corner top
9 Chamber middle bottom To know the temperature distribution in these regions
10 Chamber middle top
11 Chamber middle
12 Sterile door left corner top  

Any conduction of heat through the door, which may cause temperature drop at that particular point.

13 Sterile door middle
14 Sterile door right corner bottom
15 Steam in let left side To verify the probability of excess temperature near the steam inlet.
16 Steam in let right side
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).
  • Temperature Sensor Placement And Justification For Location Selection (Garment Load Maximum)
Load Details
12 pairs of aseptic area gowning Each one number of trunk, pair of booties and headgear in one cloth bag. Four such bags are placed side by side as one layer in the trolley and three such layers are loaded amounting to 12 pairs.
Three numbers of glove packs Each containing 8 pairs of powder free latex gloves.
12 numbers of lint free mopping dusters Wrapped using parachment paper and arranged side by side

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Inside garments pack -01 To verify the temperature penetration in the innermost portion of these packs.
7 Inside garments pack -03
8 Inside garments pack -05
9 Inside garments pack -07
10 Inside garments pack -09
11 Inside garments pack -11
12 Inside mopping dusters pack top
13 Inside mopping dusters pack bottom
14 Inside the gloves pack
15 Chamber middle top To verify the temperature distribution above the packs in the bottom rack.
16 Chamber middle bottom
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).

 

  • Temperature Sensor Placement In The Machine Parts Load
Load Details
o       Powder hopper – 1 number

o       Rubber stopper hopper – 1 number

o       Rubber stopper hopper chute – 1 number

o       Powder wheel with pistons – 10 numbers

o       1 number of 0.2-micron Catridge filter with housing for N2/ CO2 filtration

o       2 nos. of vent filters of 0.2-micron porosity for nitrogen filtration at dosing stations

o       Suitably wrapped silicone tubing & Forceps

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Powder hopper top To verify the temperature penetration in the innermost portion of these parts.
7 Powder hopper bottom
8 Index wheel port
9 Rubber stoppers hopper
10 Dust collector set
11 Rubber stoppers chute
12 Sterilizing grade filter and tubing pack
13 Accessories (forceps etc)
14 Chamber middle bottom To verify the temperature distribution at these points during sterilization cycle.
15 Chamber middle top
16 Near steam inlet
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).

 

  • Temperature Sensor Placement In The Filtration Accessories Load
Load Details
  •   293 mm diameter membrane holder previously washed with WFI and fitted with 0.2-micron N66 membrane filter and 2 micron pre-filter with suitable silicone tubing for disinfectant filtration.
  •  25-liter pressure vessel previously washed with WFI for storage of sterile filtered disinfectants.
  •  Silicon tubing required for water filtration and clamps required for fixation of the membrane holder and tanks assembly.

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Membrane holder inside top To verify the temperature penetration in the innermost portions of the loaded items.
7 Membrane holder inside bottom
8 Pressure Vessel inside top
9 Pressure Vessel inside bottom
10 Pressure Vessel inside middle
11 Inside Silicon tubing
12 Inside Silicon tubing pack
13 Near steam in let To verify the temperature distribution at these points during sterilization cycle.
14 Near non-sterile door middle
15 Near sterile door middle
16 Chamber middle
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).
  • Temperature sensor placement in the rubber stoppers load (Maximum).
Load Details
Pre-defined nos. of previously washed and siliconised 20 mm stoppers loaded in 4 nos. of SS perforated cassettes (pre-defined nos. approximately in each cassette amounting to nos.) These cassettes are loaded into the rotating carriage of the bung processor and the carriage is loaded into the chamber of the bung processor.

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Cassette 01 with stoppers To verify the temperature penetration in the innermost portions of the loaded items.
7 Cassette 02 with stoppers
8 Cassette 03 with stoppers
9 Cassette 04 with stoppers
10 Cassette 05 with stoppers
11 Cassette 06 with stoppers
12 Cassette 07 with stoppers
13 Cassette 08 with stoppers
14 Chamber middle To verify the temperature distribution at these points during sterilization cycle.
15 Near non-sterile door middle
16 Near sterile door middle
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).
  • NOTE: As it is not possible to insert the temperature sensors in the Rotating carriage of the bung processor during Sterilization and drying as the carriage rotates during the sterilization and drying process. Hence the basket drive of the equipment has been intentionally stopped and the sensors are introduced into the static carriage during this heat penetration study. This simulates worst case study of stationary load for sterilization and drying.
  • Temperature Sensor Placement In The Rubber Stoppers Holding Canisters
Load Details
Pre-defined nos. of rubber stopper holding canisters (without perforations and with lids) previously washed with WFI, used for holding washed-siliconised-sterilized-dried, rubber stoppers. Arranged pre-defined nos. in one row and 2 such rows are placed in the sterilizer chamber amounting to  nos. of canisters.

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Inside canister -01 To verify the temperature penetration in the innermost portions of the loaded items.
7 Inside canister -02
8 Inside canister-03
9 Inside canister -04
10 Inside canister -05
11 Inside canister -06
12 Inside canister -07
13 Inside canister -08
14 Near non-sterile door middle To verify the temperature distribution at these points during sterilization cycle.
15 Near steam in let
16 Chamber middle
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).

 

  • Temperature Sensor Placement In The Media Vessels with WFI Water
Load Details
Pre-defined nos. of pressure vessels (used for media) contains WFI water around 17 – 18 liters placed in the sterilizer chamber.

 

Sensor No. Location In The Chamber Justification For Location Selection
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.
2 Near RTD  -02 As these are temperature recording probes at different points. It is necessary to verify/compare the temperature distribution obtained with strip chart / print out from the sterilizer
3 Near RTD – 03
4 Near RTD – 04
5 Near RTD – 05
6 Inside vessel –1 (top level –01) To verify the temperature penetration in the innermost portions of the loaded items.
7 Inside vessel – 1 (bottom level –02)
8 Inside vessel –2 (top level –03)
9 Inside vessel – 2 (bottom level –04)
10 Inside vessel –3 (top level –05)
11 Inside vessel – 3 (bottom level –06)
12 Inside vessel –4 (top level –07)
13 Inside vessel – 4 (bottom level –08)
14 Near non-sterile door middle To verify the temperature distribution at these points during sterilization cycle.
15 Near steam in let
16 Chamber middle
  • (NOTE: The temperature sensors shall be placed in the pre determined locations with predetermined sensor numbers corresponding to the data logger channels).

 

  • Location of the Bowie – Dick Test Pack

37

  • REQUALIFICATION CRITERIA
  • Revalidation shall be carried out in case of
  • Change of cycle program
  • Inclusion of new load
  • Replacement of any major component / instrument.
  • Major modification in the existing equipment / system / utility.
  • During monitoring if equipment / system is found to be malfunctioning.
  • Shifting of the equipment / system from one location to another.
  • If any failures in Chamber leak process continuously.
  • If any failures in standard sterilization process continuously
  • If any failures in HPHV Steam sterilization process continuously
  • If any failures in Bowie and dick process continuously

 

 

 

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