VAL-225 Procedure for Performing Steam in Place (SIP) Validation Pharmaceuticals quality assurance & validation procedures GMPSOP

Department	Validation/Technical Services		Document no	VAL-225
Prepared by:		Date:		Supersedes:
Checked by:		Date:		Date Issued:
Approved by:		Date:		Review Date:

1. PURPOSE

The objective of this Standard Operating Procedure is to outline the validation approach for sterilization-in-place (SIP) process validation of all biological production area vessels, both in empty and full states, associated pipework, transfer lines, filters and manifolds at a GMP site.

2. SCOPE

This SOP details the procedure to be carried out when performing validation studies on the vessels, associated pipe work, transfer lines, filters and manifolds for the biological production area at a GMP site.

Validation will be carried out in accordance with an approved protocol or approved protocol plan test sheets. The protocol will provide detailed test procedures, assign responsibilities and define acceptance criteria.

3. RESPONSIBILITY

It is the responsibility of all validation personnel to ensure that validation activities are carried out in accordance with this SOP. Contractors carrying out validation activities on behalf of the Principal (Customer) must comply with the requirements of this document.

General responsibilities for validation activities are defined in VAL-080 Validation Master Plan.

4. SAFETY AND PROCESS SPECIFIC INFORMATION

All safety requirements for relevant areas at a GMP site must be followed at all times.

PPE required: Safety Glasses, insulated gloves, production coveralls, hearing protection when pumps are operating in the area.

– The appropriate Personal Protective Equipment (PPE) must be used when dealing with the vessels extremes of temperature.

– Wear safety glasses and disposable sterile gloves when performing studies with Biological Indicators (BIs). Ensure all BIs are returned to the QC Microbiology Laboratory for testing or appropriate disposal.

– Ensure that a work permit form “Permit to Work: Approval” form has been completed to authorize validation work on the target equipment(s). Note that to isolate utilities, an additional form “Equipment Isolation Lock-Out and Tag-Out” (Appendix A) must be completed as part of the “permit to Work: Approval” form.

– All access to vessels must be conducted following Form “Equipment Tag Out for Validation” (Appendix B)

5. GLOSSARY/DEFINITIONS

• BI – A biological indicator is a spore strip/ ampoule or inoculated item with a population of spores of no less than 106 (for strips and dry BIs) and 106 (for Wet BIs) Geobacillus stearothermophilus with known D (minimum of 1.5 minutes) and Z values.

• Control BI – A BI of the same lot as those used for testing, but one that was not exposed to the sterilization cycle which has not been treated in the same manner.

• Empty Vessel Sterilization – Sterilization of vessel without presence of liquid.

• Full Vessel Sterilization – Sterilization of vessel with liquid.

• Conditional Approval – Approval to resume production use of vessel prior to the final re-qualification report approval based on specified requirements and conditions being met.

• Equilibrium Time – The period that elapses between the attainment of the sterilization temperature of 121°C in vessel and the attainment of the sterilization temperature at all test points.

• F₀ Value – Calculated sterilization time at the current temperature and time equivalent to the sterilization time at the base temperature at 121°C.

• D-value – The D value is defined as the time in minutes at a given set of parameters required to reduce the population of a biological indicator (BI) by one log.

• Run Time – Period for which the temperature at all points within the system is held equal or above the required temperature of 121°C.

• Donor Vessel – The vessel from which the steam is supplied from during the transfer line sterilization process

6. CALCULATION THEORY

6.1 F₀ values must be calculated for every thermocouple placed inside the vessel.

6.2 The following equation must be used and entered correctly in the excel spread sheet. The formula below must be entered into each cell where the corresponding recorded temperature is equal or above 100°C:

6.3 F₀=∆t x 10^(T-121)/z

a. Where ∆t = 0.0833 minutes, (5sec/60sec = 0.0833 minutes, assuming a 5 second sampling interval)

i. z = 10^oC

ii. T = Temperature (^oC) value for that particular time point

b. Thus F₀= 0.0833 x 10^(T-121)/10

6.4 The formula to be typed into an excel spreadsheet for each individual cell is

a. =0.0833*10^((Cell Value -121)/10)

6.5 The F₀ values calculated for each cell are added together over the duration of the SIP time period to give the total FO value for that particular location.

6.6 Example:

	A	B	C
1	Time	Temperature (^oC)	Calculated F₀value
2	13:50:00	122.4	=0.0833*10^((B2-121)/10)
3	13:50:01	122.6	=0.0833*10^((B3-121)/10)
4	13:50:02	122.5	=0.0833*10^((B4-121)/10)
5	Accumulated F₀		=SUM(C2:C4)

6.7 The equation entered must be verified to be correct by a second validation engineer or delegate. The verification signature must be on the results sheet of each run in the validation report.

7. RISK ASSESSMENTS

All SIP validation activities are carried out based on the following associated Risk Assessments.

Risk Assessment No.	Description
122004	Risk Assessment for SIP’s of all Blending and Filling Tanks
122156	Risk Assessment for the SIP of all Antigen Tanks
102038	Blending Area Transfer Line SIP Risk Assessment
102093	Risk Assessment – Sterilization of Filling Lines

8. VALIDATION RATIONALE

8.1 Validation Rationale
Sterilization In Place Performance Qualifications and Re-qualification are required in the following circumstances:

• New Equipment, including Vessels, associated pipe work, transfer lines, filters or manifolds
• Changes to the SIP procedure or parameters
• Major changes to vessels, associated pipe work, transfer lines, filters and manifolds as directed by a change control
• Scheduled compliance

8.2 Revalidation Approach
Vessels, transfer lines, filter systems or manifolds used in the biological production area must be re-validated as per the re-validation schedule. For the purposes of re-validation all the vessels are classified as groups based on the volume, internal diameter and the number of connections on the vessels. Only one vessel from each group is to be revalidated. An example of “Schedules and vessel groupings are detailed in Appendix C.

Vessels subject to Department of Agriculture bio-security quarantine rules must be re-validated on an annual basis.

Before the commencement of re-validation activities a review of all previous validations completed on the chosen vessel, transfer line, filter system, intermediate process equipment or manifold shall be conducted.

9. PROTOCOL PREPARATION

9.1 PQ Protocol Preparation

Protocols for all SIP validation depending on the equipment type are to be raised by completing Appendix D “Sterilization- In- Place: Validation Protocol Plan Approval” alongside the following documents for the respective SIP execution type:

Equipment Type	Document Number	Document Title
Vessels	Appendix E	Sterilization In Place – Empty/Full Vessel Protocol
	Appendix F	Sterilization In Place Validation – Vessel Diagram
	Appendix G	For new equipment or cycles Sterilization In Place – Data Collection Table
Transfer Lines	Appendix H	Sterilization In Place – Transfer Lines Execution
	Appendix I	Sterilization In Place Validation – Transfer Line Diagram
	Appendix J	For new equipment or cycles Sterilization In Place Validation – Transfer Lines Data Collection Table
Manifold	Appendix K	Sterilization In Place Validation – Antigen Transfer Manifold

• Alternatively, where justified, a validation protocol may be raised for new processes or significant changes to a pre-existing SIP processes.

• A review of critical documentation, maintenance and calibration and where relevant a review of previous standard operating procedures (SOP) as per VAL-265 “Critical Documentation Verification During a Validation Study”.

• Obtain standard vessel or equipment SIP times from the applicable corresponding SOP. If applicable, review the Department of Agriculture (DoA) Permit for any specific sterilization conditions (e.g. F¬0, Temp.) for Antigen area equipment.

• A clear description of the of run type (full or empty, maximum, minimum or the worst case volume of liquid to be sterilized, DoA requirements).

• Description of the external monitoring points, thermocouple and Biological Indicators placements inside the vessel. A thermocouple must be placed at the temperature transmitter port or other suitable worst case location for every run completed. Determination of the locations is based on the risk assessments completed. Refer to the individual risk assessments completed on the vessels or pipe-work configuration to be validated.

• Recording device must have a recording accuracy of at least 0.8°C and capable of recording data every 5 seconds.

• The protocol should specify the number of planned runs to be completed for both the empty and/or full states for vessels.

• If no prior validations have been conducted then a total of three successful runs are to be completed for each state, more specifically three empty runs, as well as two runs on the minimum working volume of the vessel and one run on the maximum working volume of the vessel. For the purpose of revalidation, the number of runs may be reduced to one run at the empty state and one run at the filled state. For the filled state, the volume chosen shall be alternating between the maximum and minimum working volume for each revalidation cycle

• For transfer lines and manifolds, steam flow from both direction where practical should be validated if no prior validation have been conducted. Two runs is required using the direction of the standard process steam flow, and one run with the steam flow in the reverse direction to challenge the process. If the reverse flow is not practical or possible then three runs shall be conducted using the standard flow direction. For the purpose of re-validation only one run is required to be completed using the standard direction of flow.

• If the number of runs is to be increased or reduced from the number of runs stated in the initial protocol, a justification must be made in the final validation report for the change in the number of planned runs.

• The use of half cycle times will be employed for transfer lines, manifolds and empty vessel runs. While the full cycle times will be employed for all filled vessel runs. A justification must be made in the final validation report if any changes are made to the cycle times used for any vessel validated.

9.2 Thermocouple and Pressure Transducer requirements

• For heat distribution data, thermocouples must be evenly distributed inside the vessel or transfer lines to map the temperature at the critical locations based on the risk assessment completed.

• A minimum of six thermocouples are to be used for vessel SIP and three for transfer line and manifold. A minimum of two extra thermocouples should be prepared for replacement in case of damage or faults.

• During placement of the thermocouples, where possible ensure that the thermocouples are not contacting metal surfaces (internal walls). Care should be taken not to over tighten bolts and gaskets.

• Description of the vessel or system being studied with sufficient detail on thermocouples and BI placement within the vessel (record the details in form Appendix F), or transfer lines (Appendix I). The placement of BIs must be completed in accordance with VAL-200 (Selection and Use of Biological Indicators during Validation Studies).

• A pressure transducer should also be placed at a suitable location for the continuous monitoring of pressure. Area steam supply pressure readings from DAS software display (iFix) should be recorded manually for the start, middle and end of the run.

10. EXECUTION OF THE PQ PROTOCOL

10.1 A half cycle for any empty vessel and a full cycle for any filled vessel is employed, unless otherwise stated. Review the vessel operation SOP to determine the current validated sterilization time period(s).

10.2 Synchronize the times of the Graphtec or Eurotherm Data loggers with the DAS time.

10.3 The graphic recorder shall be set to record data in 5 second intervals.

10.4 Place the thermocouples and biological indicators according to the locations selected on form Appendix F or Appendix I diagram.

10.5 Ensure that the vessel and fittings are closed and that all of the bolts have been tightened due to the placement of the thermocouples.

10.6 Ask an area operator to inspect the integrity of system access points.

10.7 If a full vessel trial is required fill the vessel with the required amount of chosen solution in accordance with the relevant procedure stated in the SOP.

10.8 Unless specific training has been completed for operation of the vessel or equipment for the validation personnel engaging in the study, qualified area personnel must conduct vessel or equipment operation during the SIP evaluation process.

10.9 Initiate the sterilization of the vessel, transfer lines, or manifold according to SOP. During the empty vessel run the inlet filter is sterilized prior to the sterilization of the tank and associated pipe work.

10.10 However during the validation of the full vessel, only the tank and associated pipe work are required to be sterilized, as the inlet filter sterilization process was completed during the empty vessel runs.

10.11 If the validation is for a new piece of equipment or process then also document external monitoring point temperatures, pressure inside the vessel or lines and input of (LP) steam every 5 minutes of the run using the form Appendix G or Appendix J Data Collection Table.

10.12 Once the sterilization process has been completed, cool the vessel and/or transfer lines down in accordance with the relevant procedure stated in the SOP

10.13 After a successful run, retrieve the BIs and send them to the QC microbiological laboratory for testing. Fill out the relevant form for the relevant type of Biological indicator. (Appendix N EZ Test Crushable Ampoules, and/or Appendix M MagnaAMP Glass Ampoules/SterilAmp II Glass Ampoules, and/or Appendix L MesaStrip Spore Strips)

10.14 Retrieve the data from the graphic recorder and compare the result to the acceptance criteria. Attach a copy of summarized data to the final validation report Appendix K.

11. ACCEPTANCE CRITERIA

Acceptance criteria for all SIP studies shall be based on:

• A minimum accumulated F0 of 22 minutes and a minimum temperature of 121°C must be achieved at all thermocouple locations. A minimum F0 of 15 minutes is considered acceptable for heat sensitive liquids unless subject to specific Department of Agriculture requirements, however a justification must be made in the final validation report.

• All temperature monitoring locations during the holding period are maintained at a minimum temperature of 121°C.

• Total kill of all BIs, except for the positive controls which must show growth.

• DoA requirements if any have been satisfied.

• At the mid point of the sterilization phase the mean value of all the recorded temperatures measured over a 1 minute period is within ±2°C of the corresponding saturated temperature calculated using the pressure reading from the pressure transducer over the same time period (as per BS EN285:2006). For example, for a 15 minute sterilization evaluation, the reported values from the 7 – 8 minute time period would be utilized for the saturated steam condition evaluation.

• A minimum of 80% of thermocouples used in the run must meet the post run verification acceptance criteria in order for a run to be successful. However thermocouples located as close as practicable to the controlling temperature sensor or known worst case location must also meet post calibration as part of the acceptance criteria for a successful run. These are to be identified in the diagrams and/or summary report.

• If any of the above tests fail to meet the acceptance criteria, complete a detailed deviation report. Upon resolving the cause of the failure the run can be repeated if the nature of the failure does not warrant the closure of the study and the re-issuing or amendment of the protocol.

12. COMPLETION OF THE VALIDATION REPORT

A SIP validation report form, Appendix K shall be completed for each run, it will include:

• An evaluation of the data.
• A comparison of the results to the procedural steps and parameters utilized.
• Documentation of the maximum and minimum temperature, pressure and F₀, where appropriate, from the exposure period of each run.

There shall be confirmation that SOP(s) used in the execution of the PQ for the successful runs are finalized and approved as per VAL-265 “Critical Documentation Verification During a Validation Study”. Conclusions based upon the execution of the study shall be incorporated into the SOP(s) applicable to the usual sterilization practices.

The ability of the steam sterilization process to inactivate micro-organisms to the required level shall be demonstrated by the completion of the validation and by the approval of the validation report.

12.1 Conditional Release

A vessel, transfer line, filter system or manifold may be released for the use of production use prior to final approval of the validation report where a conditional release is granted.
The Validation Department is responsible for issuing the conditional release document (Appendix O Conditional Release Form) for QA approval.

Conditional Release may only be provided where thermometric and barometric run data has been evaluated and been shown to meet the acceptance criteria.

Appendix A: Lock Out Tag Out Form

Appendix B: Equipment Tag Out for Validation

Appendix C: Rationale and Schedule for Steam in Place (SIP) Validations on Tanks, Transfer Lines, Filters, and Manifolds – Example

Appendix D: Sterilization in Place: Validation Protocol Plan Approval

Appendix E: Sterilization in Place – Vessel Execution Record

Appendix F: Sterilization in Place Validation – Vessel Diagram

Appendix G: Sterilization in Place Validation– Data Collection Table

Appendix H: Sterilization in Place Validation– Data Collection Table

Appendix I: Sterilization in Place Validation – Transfer Line Diagram

Appendix J: Sterilization in Place Validation – Transfer Lines

Appendix K: Sterilization in Place – Validation Summary Report

Appendix L: Biological Indicator Test Results Sheet – Spore Strips