Protocol for re-validation of dry heat sterilizer
Equipment Name : Dry Heat Sterilizer
Equipment ID :
Protocol Approval Date :
No Of Pages
|Point No.||Section Title||Page No.|
|—-||Protocol cover page|
Protocol contents and List of Annexure
|2.0||Validation Team & Responsibility|
|3.2||Purpose and Scope|
|3.4||Training and Validation|
|5.0||Requirements for validation|
|7.0||Procedure for re-validation|
|9.0||Evaluation and approval of validation report|
|11.0||Change Control System|
List of Annexure
|Sr. no.||Annexure number||Title||Number of pages|
|1||Annexure – I||Evaluation and approval of validation report|
|2||Annexure – II||Data logger calibration certificates|
|3||Annexure – III||Bacterial Endotoxin Indicator Recovery And Log Reduction|
|4||Annexure – IV||Schematic diagram for Probe locations and endotoxin challenge vials|
|5||Annexure – V||Temperature mapping summary|
|6||Annexure – VI||Training Record|
1.0 Protocol Approval
This is a specific protocol for Re-validation of Dry Heat Sterilizer (Dry heat sterilizer Equipment ID No.: -…) located in the microbiology area- Instrument Room. Signing of this approval page indicates agreement with the validation described in this protocol. If any modification to this procedure is necessary, during execution, an addendum shall be prepared and approved.
2.0 Validation Team and Responsibilities
Microbiology Microbiology department shall be responsible for preparation, execution and perform the validation activity as per protocol, after that completion compile summary report.
Microbiology Head shall be responsible for the review of protocol and report , validation requirements and regulatory requirements, testing & result
Engineering Engineering department shall be responsible for:
Checking of protocol to ensure the availability of utility, instruments, and equipments, calibration of all gauges, indicators, instruments and verification of preventive maintenance.
Quality Assurance Executive- He shall be a member of validation team and responsible for execution of the DHS validation. He shall also be responsible for compilation of validation reports.
Head-QA or nominee of validations shall be a member of execution team and shall be responsible for supervision of validation activities in accordance with this protocol. He shall also be responsible for review of validation reports.
Head–QA shall be responsible for approval, after verification the protocol for its correctness and compliance as per regulatory requirements. Head-QA shall also be responsible for training of validation team.
Validation Team Department Name Designation Sign Date
The objective of this protocol is to validate the periodically of Dry Heat Sterilizer (Equipment ID No.: located in the Microbiology Lab.
The Dry heat sterilizer is used for the depyrogenation of BET assay tubes, sampling scoops, and glass vials. This requires ensuring that the depyrogenation process is adequate, suitable and effective to provide necessary depyrogenation.
For this a validation study shall be carefully designed so as to provide a high degree of assurance that the depyrogenation process employed is capable of repeatedly providing the desired depyrogenation assurance level.
The validation study shall consist of heat penetration study challenged with endotoxin indicator challenge in maximum load pattern to be depyrogenated in this Dry heat sterilizer. The purpose of this testing shall be to verify the depyrogenation of the load pattern, when the established critical parameters fall within the established range of acceptability.
3.2 Purpose and Scope
The Purpose of this re-validation Protocol is to establish documentary evidence to ensure that the procedure followed for depyrogenation of articles in Dry heat sterilizer is capable of achieving:
– Greater than 3-Log reduction in challenged heat resistant Bacterial Endotoxin in the Loaded Chamber.
The scope of this re-validation is limited to depyrogenation of articles in the Dry Heat Sterilizer Equipment No.:
3.3 Validation Approach
The selected validation approach is periodic validation.
3.4 Training and Validation
Section in-charge / Head-QA/ designee shall give the class room training to the execution team on total validation activities.Training record shall be recorded in Annexure No.: VI.
4.0 Reference Documents
5.0 Requirements for validation
5.1 Operate Dry Heat Sterilizer as per their respective Standard operating procedure (SOP No. ) The validation study follow as per bellow:
6.0 Equipment Introduction
Dry heat sterilizer (DHS) is using for depyrogenation of glass and stainless steel accessories. DHS has one blower to circulate heat and air in the chamber. Chamber has three partitions though three porous stainless steel trays. Heating is provided by heating coils. Blower maintains the inner air circulation and prevents the chamber from outside non-sterile air. The volume of chamber is aprox 90 liter.
Set temperature of DHS is 200 °C for hold time one hour duration. In this duration all placed items as per load pattern got depyrogenation.
7.0 Procedure of re-validation:
7.1 Basic Equipment Validation Checks:
7.1.1 Prior to start periodic validation activity, Dry heat sterilizer shall be checked for the correct and sequential procedure of its basic instrumentation, to ensure that all the essential components are functioning properly.
7.2 Calibration of External Data Logger Probes:
7.2.1 The Eight or more than eight probes data logger shall be used for the temperature measurements of dry heat sterilizer.
7.2.2 Deviation is not more than 0.5 ‘C from the set temperature.
7.2.3 The pre and post calibration of Data logger probes shall be done during validation study. Certificate of pre and post calibration shall be attached with temperature mapping report.
7.2.4 Detailed calibration report derived from data logger [Pre and post] shall be attached to the Validation Report as Annexure-II.
7.3 Preparation and quantification of Endotoxin Challenge Vials [ECVs]
7.3.1 Endotoxin Challenge Vials [ECVs] to be used as an Endotoxin indicator shall be prepared in the Microbiology laboratory, using standard Endotoxin Indicator [EI] having 100,000 EU per vial of Escherichia coli Endotoxin. The Certificate of Analysis [COA] provided by the vendor shall be attached to the validation report.
7.3.2 The Endotoxin indicators shall be spiked in assay tubes [10×75 mm / 12×75 mm] and 20 mL glass vials.
7.3.3 Take 08 number of Endotoxin indicator vials containing 100000 EU/vial. Reconstitute each vial with 1 ml of LAL reagent water and vortex as per time given in the certificate. Dispense 0.1 ml aliquots of reconstituted Endotoxin indicator in de-pyrogenated vials (use the same sized vial which is used in validation), so that each vial will contain 10000 EU. Keep the spiked vials open under LAF over night for drying. Visually observe for drying and label the vials with spiked concentration, date of preparation and sign. Close the vials with aluminium foil, until they expose to validation cycle. Use the prepared vials within 2 days. Record the preparation details in Annexure –III.
7.4 Temperature monitoring equipment and devices.
7.4.1 A pre-calibrated external multi channel data logger, capable of displaying and recording temperature with a sensitivity of 0.1 C, shall be used for recording the temperature readings at different locations within the Dry heat sterilizer chamber.
7.4.2 The external multi channel data logger having flexible temperature sensors, capable of sensing temperature with a sensitivity of 0.1 C shall be employed.
7.4.3 The external multi channel data logger having an in-built real-time clock, to record temperature at predetermined time intervals shall be employed.
7.5 Validation Methodology
Validation studies shall be done in following steps,
7.5.1 Empty Chamber Heat Distribution [ECHD] study:
One cycle shall be performed for ECHD study without any change in procedure.
7.5.2 Loaded Chamber Heat Penetration [LCHP] study:
One cycles of LCHP shall be performed for maximum loading configurations (if there is no any change in procedure).
7.6 Empty Chamber Heat Distribution study
7.6.1 Empty chamber heat distribution shall be performed to ensure that the achieved dry heat temperature is uniformly distributed to the entire chamber during depyrogenation hold time with predefined temperature.
7.6.2 Following approach shall be followed for Empty Chamber Heat Distribution [ECHD] study:
7.6.3 ECHD cycles shall be performed with depyrogenation temperature range of 185-215°C [200±15°C] for 60 minutes of depyrogenation hold period, using external multi channel data logger temperature sensors and in-built temperature sensor of the Dry heat sterilizer.
7.6.4 12 external probes in the chamber shall be placed at predefined locations as mentioned in the Annexure-IV. These probes shall be placed in such that all the probes are evenly distributed through the chamber.
7.6.5 It shall be ensured that the temperature sensors do not touch any metal surface or walls of chamber.
7.6.6 The equipment shall be operated as per the operating manual or as respective SOP on ‘operation, maintenance and cleaning of dry heat sterilizer, SOP no.
7.6.7 After completion of the cycle, the print outs of data logger shall be collected and reviewed for the data and the observations shall be recorded in the Annexure-V.
7.6.8 The cycle print outs of the data logger and equipment shall be attached with report.
7.6.9 Total performed cycles shall be studied. Qualify the Empty Chamber Heat Distribution prior to Loaded Chamber Heat Distribution studies.
7.6.10 The in-built temperature sensor shall be monitored for the temperature behaviour, as routine cycles are being controlled based on these temperature sensor.
7.6.11 Acceptance Criteria
The temperature readings recorded by both the external multi channel Data Logger and in-built sensors, must be maintained within 185-215°C [200+15°C] for 60 minutes of depyrogenation hold period.
7.7 Loaded Chamber Heat Penetration Study :
7.7.1 Heat Penetration studies shall be carried out for each load type by challenging with bacterial Endotoxin Challenge Vials [ECVs] to demonstrate adequate heat shock available to the items under depyrogenation for achieving greater than 3-Log reduction in bacterial endotoxin.
7.7.2 Following shall be the approach for Loaded Chamber Heat Penetration Study.
7.7.3 Loaded Chamber Heat Penetration cycles shall be performed with depyrogenation temperature range of 185-215°C [200±15°C] for 60 minutes of depyrogenation hold period, using external multi channel Data Logger temperature sensors and Endotoxin indicator challenging vials.
7.7.4 The articles shall be loaded into chamber as per the loading pattern described in Table-I and 12 external data logger probes shall be placed in the chamber at predefined locations as per Annexure – IV.
7.7.5 Endotoxin Challenge Vials [ECVs] challenged during all cycles of Heat Penetration shall be with a preload of 10,000 EU Endotoxin and the indicator shall be placed inside the load.
7.7.6 Minimum 8 No of ECVs shall be challenged for each cycle.
7.7.7 The Endotoxin indicators shall be the spiked in assay tubes [10×75 mm, 12×75 mm] &20 mL glass vials.
7.7.8 The Endotoxin indicators shall be placed inside the respective packs as a simulation for depyrogenation.
7.7.9 The locations of the Endotoxin indicators shall be as depicted in Annexure-IV and the specific numbers shall be as per the Table-II.
7.7.10 The locations for the placement of the temperature sensors and ECV’s shall be maintained same for each cycles to check the reproducibility of temperature profile and log reduction in bacterial Endotoxin.
7.7.11 After completion of cycle, collect the Endotoxin indicator vial and tested for BET.
7.7.12 After the depyrogenation cycle, the challenged endotoxin indicator vials shall be tested using Gel Clot Assay Method as per the current GTP on “Endotoxin test by gel clot method” (GTP/023).
7.7.13 The test results shall be recorded in the Annexure-III.
7.7.14 After completion of the cycle, the print outs of data logger and equipment shall be collected and reviewed.
7.7.15 The copies of the test report shall be attached to the validation report as Annexure V.
7.7.16 All the challenged vials & assay tubes must exhibit greater than 3-Log reduction in Bacterial Endotoxin level.
|Load||Description||Minimum Load||Maximum Load||Pack|
|Assay tubes||10 x 75 mm [10 per pack]||50||250||25|
|Assay tubes||12 x 75 mm [10 per pack]||50||200||20|
|Assay tubes||10 ml capacity 10 per pack||20||100||10|
|Glass vials||20 mL capacity [20 per pack]||20||100||10|
|SS sampling scoop||Individual||1||5||–|
|Endotoxin Indicator number||Spiked in|
|EI-01||10 x 75 mm|
|EI-02||10 x 75 mm|
|EI-03||12 x 75 mm|
|EI-04||12 x 75 mm|
|EI-05||12 x 75 mm|
|EI-06||20 mL vial|
|EI-07||20 mL vial|
|EI-08||20 mL vial|
7.7.17 Acceptance Criteria
The temperature readings recorded by both the external multi channel Data Logger and in-built sensors, must be maintained within 185-215°C [200 +15°C] for the depyrogenation hold period of 60 min.
Endotoxin challenge vials after depyrogenation, must exhibit NLT 3-Log reduction [Initially having 10,000 EU/vial of Escherichia coli endotoxin], when tested Gel Clot Assay Method.
7.8 Action to be taken in case of Deviation / Changes / Failure
Any deviation / changes / failure occurred during execution of the protocol, shall be addressed in the validation report. Accordingly recommendations shall be made in the validation report. Investigation shall be carried out and suitable corrective / preventive actions shall be taken.
8.0 Re-validation Criteria
Dry heat sterilizer shall be re qualified incase of,
Change in the configuration of the loading pattern.
Any major change in the equipment/ major breakdown.
Three cycles of ECHD shall be performed (if any change in procedure).
Three cycles of LCHP shall be performed for maximum loading configurations (if any change in procedure).
9.0 Evaluation and Approval of Validation Report
The report shall be evaluated and proper references / conclusions / recommendations shall be recorded by validation team.
The validation report shall be finally approved by Head-QA.
|No.||Number||BET||Bacterial Endotoxin Test|
|SOP||Standard Operating Procedure||GTP||General Test Procedure|
|PQ||Performance Qualification||IPA||Isopropyl alcohol|
|COA||Certificate of analysis||QA||Quality Assurance|
|LAL||Limulus Amoebocyte Lysate||EU||Endotoxin unit|
11.0 Change Control System
The changes in the proposed / system shall be carried out through the change control system.
12.0 Revision History
|Revision No.||Date of Revision||Change Control No.||Reason for Revision||Superseded Document No.|