Design Requirements for Facilities and Equipment and Their Qualification – A Case Study

Patrick J. McCormick, Ph.D. pmccormick@bausch.com + 1 585 338 8390
Photo courtesy of Texwipe

Pyrogens and Endotoxin • Sterilization and depyrogenation seldom drive the development of new products, but are essential to the safety of our products and the health of our customers.
Endotoxin tied to sterilizers. Hospital traces fatal outbreak. Saturday, December 16, 1995.

Cookson, S.T. et al. Pyrogenic Reactions in Patients Undergoing Cardiac Catherization Associated with Contaminated Glass Medicine Cups. Catherization and Cardiovascular Diagnosis 42:12-28(1997) Outbreak traced to enzyme detergent contaminated with > 104 CFU/ml; 434 EU/ml; reprocessed cups had 2,250 EU.

Pyrogens and Endotoxin • Pyrogen: any substance that induces a fever. • Exotoxin: soluble protein secreted by microorganisms that is toxic to cells (botulism toxin, diphtheria, toxic shock syndrome, etc.). • Enterotoxin: soluble protein secreted by microorganisms in the intestine (food poisoning). • Endotoxin: high MW complex associated with the cell wall of G (-) bacteria that is pyrogenic in humans and specifically interacts with LAL.

Pyrogens and Endotoxin Endotoxin: • Threshold pyrogenic response of 1 ng/kg where 5 EU is equivalent to 1 ng E. coli (EC-2) reference endotoxin. • Complex host response (inflammatory cytokines) can lead to high fever, severe tissue damage and death. Depyrogenation: • Validated process designed to remove or inactivate endotoxin (ANSI/AAMI ST72:2002).

Pyrogens and Endotoxin • For most medical devices an endotoxin limit of 20 EU/device is generally recognized, although some medical devices such as IOLs may have lower limits (ISO 11979-8; 2 EU/IOL). Consult applicable regulatory guidelines (ISO, EN, FDA).

– 0.2 EU/kg/hr – intrathecal. • Consult pharmacopeias and applicable regulatory guidelines. .Pyrogens and Endotoxin • For most pharmaceutical applications the generally accepted endotoxin limit varies with the route of administration: – 5 EU/kg/hr – intravenous.

0001 0. • FH at 170 oC.1 0. 1% pos. 1000000 100000 10000 1000 100 10 1 0. • Total kill of 106 spores of Bacillus atrophaeus at half cycle based on overkill approach of ISO 14937/14161.001 0.Dry Heat Sterilization .01 0. • z = 20 oC.00001 0.Kinetics • Well defined 1st order inactivation kinetics. exposure 10-6 SAL .000001 1/2 cycle window 63% pos.

Kinetics The successful validation of depyrogenation is generally accepted as evidence of the successful validation of dry heat sterilization due to the much greater resistance of endotoxin to dry heat as compared to bacterial spores.Dry Heat Depyrogenation . . Dry heat exposure* 120 minutes 60 minutes 30 minutes Sterilization 160 oC 170 oC 180 oC Depyrogenation None None 250 oC * Actual exposure time will vary with load and conditions.

J.J.Dry Heat Depyrogenation . et al. 1978. M. JPDA 36(1):23-27. – Tsuji. atrophaeus) typically are not employed. Environ.a three log reduction of endotoxin is generally accepted as evidence of successful depyrogenation and sterilization. S. – Akers.Kinetics • Endotoxin inactivation kinetics are 2nd order as opposed to first order (sterilization). . Microbio. 1982. Biological indicators (B. • Empirical validation .K. and Harrison. 36(5):710-714. Appl.

Equipment Heat transfer: • Conductive: – Heat transfer via direct physical contact.Dry Heat Depyrogenation . • Convective or radiant: – Heat transfer via fluid medium (liquid or gas). . – Air flow/shadowing effects/HEPA filtration. • Radiation: – Heat transfer via electromagnetic radiation (infrared or microwave).

Equipment Batch sterilizer: • Widely used in industry. • Simple. and infrequent use. • Ideal for varying load types. • Fans circulate heated air throughout load. small loads. rugged design.Dry Heat Depyrogenation . PDA Technical Monograph #3 .

PDA Technical Monograph #3 . flame sterilizers. • Airflow opposite product flow. infrared.Equipment Tunnel sterilizer: • Operates continuously.Dry Heat Depyrogenation . • Ideal for large load sizes. • Variable heating zones. • Fast processing time. • Convection.

etc. . filling and capping machines. Select oven and validate depyrogenation cycle time appropriate to overall manufacturing process. packaging equipment. to be realized such that uninterrupted processing may be achieved? • Total process throughput is tied to rate limiting step.Dry Heat Depyrogenation – Facilities Considerations • Integration / Process flow – How is the integration of depyrogenation oven with other equipment such as vial washers.

.Dry Heat Depyrogenation .Facilities Considerations • Capacity – Does the oven have sufficient capacity to meet current and future processing needs? • Classification – Is the classification of the oven compatible with the classification of the area where it will located and the overall manufacturing process? Most depyrogenation ovens are rated Class 100 and should therefore be suitable for application in vial and equipment prep areas of aseptic manufacturing facilities.

water.Dry Heat Depyrogenation – Facilities Considerations • HVAC / Utilities – Are current HVAC controls and utilities (electrical. air) adequate to meet the needs of the depyrogenation oven? How much added stress will be placed on the existing HVAC system and utility supply and what impact will this have on the operation of other equipment ? • Installation – Are the dimensions of the oven such that it can be transported to and installed in the desired location or will alterations to the facility and existing layout be necessary? .

Dry Heat Depyrogenation – Facilities Considerations • Building Codes – Are there any particular considerations with regard to local or regional building codes such as the need for seismic anchoring or electrical grounding requirements? • Workplace – Is the unit quiet during operation or will acoustic protection be necessary? What about additional heat stress and vibration? How will this affect production operators or other manufacturing equipment? .

Dry Heat Depyrogenation – Facilities Considerations • Maintenance / Calibration / Validation – Does the facility have adequate resources to support the maintenance. and validation needs of the oven or are additional staff and equipment needed? How to plan for interventions? • Change Control – Is the design of the oven such that it can be readily relocated should the layout of the facility be changed? . calibration.

Dry Heat Depyrogenation – Equipment Considerations • Control systems – State of the art. • Control systems – Operator interface and datalogging capability. Compatible with controls systems of other manufacturing equipment? This may be a problem if your washer and filling equipment is 20 + years old. How will critical cycle documentation be captured and stored? Accuracy and precision of recorders? Backup? What about EMF/RFI interference issues? .

Dry Heat Depyrogenation – Equipment Considerations • Pressure Balance – Is the airflow of the oven such that the appropriate pressure balance can be maintained so that product moves to progressively “cleaner” zones and classification of the surrounding areas is not impacted? • Air pathways – Are there provisions for the sterilization of air pathways within the oven. particularly the cool down zone? How is air quality of the area to be maintained during repair and maintenance of the oven? .

Dry Heat Depyrogenation – Equipment Considerations • Materials of construction – 304L SS typical with gauge and finish appropriate to application and loading conditions. . • Door interlocks – Appropriate to product flow with a safe locking sequence in the event of power loss (prevent product from exiting chamber without being adequately depyrogenated).

etc. material shed from moving parts. airflow. • Contamination – Is the oven a potential source of particulate (filter binder material.Dry Heat Depyrogenation – Equipment Considerations • Alarms – Low/High temperature. belt speed. emergency stop. etc.) or chemical contaminants (lubricants) during routine operation? Is the use of DOP a concern during validation of HEPA filters? .

Dry Heat Depyrogenation – Case Study • Retisert™ intravitreal drug implant for treatment of chronic non-infectious uveitis affecting the posterior segment of the eye. . • Uveitis is a leading causes of blindness in the Western world.

• Depyrogenation validation performed in association with Autocal Ireland Ltd and Microchem Labs. .Dry Heat Depyrogenation – Case Study Retisert implant: • Manufactured by Bausch & Lomb in Waterford. Ireland.

• Depyrogenation at 250 + 15 oC for NLT 60 minutes of fixtures and tooling in contact with product. . • Terminally sterilized by gamma irradiation (minimum 25 kGy) as per EMEA Decision Trees.Dry Heat Depyrogenation – Case Study Retisert implant: • Manufactured under cleanroom conditions.

• The move to a rigorously controlled manufacturing environment may decrease the need for extensive depyrogenation as compared to the preceding development phase. • Assess each step of process to identify critical risk areas for depyrogenation (FEMA/HACCP).Dry Heat Depyrogenation – Case Study • Depyrogenation strategy included as part of overall Cleaning Validation Strategy and Assessment. • Important to bridge development phase in order to maintain continuity of approach. .

• Must allow access for maintenance and calibration. • Door lock sequencing including power failure.Dry Heat Depyrogenation – Case Study Oven considerations: • Double door unit (Wash in / Wash out). • Must be consistent with area classification. • Size of depyrogenation oven was a major factor due to existing space restrictions. . • Oven must interface with equipment prep area to ensure effective HACCP and manufacturing flow. • Must be compatible with existing HVAC/utilities.

• Horizontal flow of heated Class 100 HEPA filtered air.Dry Heat Depyrogenation – Case Study Gruenberg L55H8.2 m3).3 ft3/ 0. • Chamber exterior: – 68” x 44” x 70”.com . • Heat exchanger (cold water) for cool down. • Bio sealing flange.epsovens.3PTSS: • Chamber interior: – 20” x 30” x 24” (8. http://www.

3PTSS: • Temperature range: – Max. http://www.thermalproductsolutions. – Operating 250 oC.5 oC/min.Dry Heat Depyrogenation – Case Study Gruenberg L55H8. • Temperature uniformity: – + 5 oC at 250 oC.com . 280 oC. – Ramp rate > 1.25% of scale at 250 oC. • Temperature Control: – 0.

USP <1211> Sterilization and Sterility Assurance. PDA TR 7 – Depyrogenation. . ISO 14937 – General requirements for sterilization. revision 2008. HTM 2010 and other regional requirements.Dry Heat Depyrogenation – Validation • • • • • • • • • EP 5.1. PDA TR 3 – 1981 Dry Heat Processes for sterilization and depyrogenation. AAMI ST63 – 2002 Dry Heat sterilization. EMEA: Decision Trees for sterilization.1 General Texts on Sterility. FDA Aseptic Processing Guidelines 2004.

.Dry Heat Depyrogenation . • Proposal from vendor (including drawings).Validation Documentation: • Validation Master Plan. • Request for Quote (RFQ). • User Requirements Specification (URS). • Validation Documentation (IQ/OQ/PQ). • Factory Acceptance Test (FAT). • Functional / Design Specifications.

• Annual revalidation – One ea.Dry Heat Depyrogenation – Validation General Validation Strategy: • Factory Acceptance test (FAT). OQ and PQ cycle. • Reduced Cycle testing – 1 cycle. • Performance Qualification Testing (PQ) – 3 cycles. • Installation Qualification (IQ). • Cycle Development / Heat Penetration (HP) studies. • Operational Qualification (OQ) – 3 cycles. .

• Verify proper ∆ P across HEPA filters. • Conduct heat distribution test (empty chamber). • Verify access ports for particle testing.Dry Heat Depyrogenation – FAT Factory Acceptance Test: • Verify physical dimensions of unit. • Verify loading system operates correctly. . • Conduct load test. • Personnel training in operation of unit.

” ISO 14937:2002. • Do we have all the parts? • Will the unit fit into the allotted space? • Do we have the necessary utilities? • Is the unit to spec? .Dry Heat Depyrogenation – IQ Installation Qualification (IQ): • “Obtaining and documenting evidence that equipment has been provided and installed in accordance with its specification.

water. • Environmental and Safety inspection. . etc. • Insulation. • Seals and gaskets. • Utilities (air. • Spare parts list. • HEPA filter installation • Baffles and panels. electrical. • Leveling/anchoring.Dry Heat Depyrogenation – IQ Confirm physical installation to specifications: • Positioning of unit. • Manuals and drawings. and certification. • Ducting and plumbing.).

• High limit alarm. . • Recirculation pressure. Calibration equipment: • Tachometer. • Exhaust pressure. • Temperature calibrator.Dry Heat Depyrogenation – IQ Verify Calibration: Oven Instrumentation: • Temperature control. • Intake pressure. • Thermal anemometer. • Chamber pressure. • Data logger. • Pressure calibrator. • Thermocouple bath. • Particle counter. • Chart recorder.

New filters should be put in place once the unit has been received and “burned in” at the maximum operating temperature as per the manufacturer's recommendation. . Do this over the weekend to minimize disruption to others. This can create an odor and haze in the air.Dry Heat Depyrogenation – IQ • Note: HEPA filters are fragile and should not be shipped installed in the unit.

” ISO 14937: 2000.Dry Heat Depyrogenation – OQ Operational Qualification: • “Process of obtaining and documenting evidence that installed equipment operates within predetermined limits when used in accordance with its operational procedures. • Does the unit perform as promised? • Let’s take it for a test drive… .

• Verify electronic backup of same software version provided on permanent magnetic media. .Dry Heat Depyrogenation – OQ Verification of PLC software: • Generate Control Software Qualification Report. • Verify hard copy of PLC ladder/logic provided. • Verify correct software version number.

Dry Heat Depyrogenation – OQ Operational Set-Up Checks: • Verify speed controller set as per manual. . • Verify high-limit controller set as per manual. indicators. • Test controls. etc. • Verify circulation fan rotating in correct direction. • Verify fan operating at correct rpm (tachometer). safety devices. • Verify chart recorder set as per manual. alarms.

= 244. .61 oC • Place 12 TC’s as indicated w/in 8 cm of chamber surface. • 20 min. • Verify temp stability Hot spot (5) = 247.Dry Heat Depyrogenation – OQ Empty chamber test: Avg. • Oven set at 250 oC.36 oC 250 oC + 15 oC (USP <1211>) over a 60 Cold spot (10) = 241.86 oC minute period. stabilization.

• Airflow measured with thermal anemometer. • Intake airflow measured at exhaust duct during process and cool down phases.Dry Heat Depyrogenation – OQ Air Velocity and Volume: • Testing performed at ambient temperature. • Performance to Manf. • Circulation airflow measured at 5 equally spaced locations on supply duct and exit duct walls approx.’s specifications. 15 cm. CFM = Average ft/min x area (ft2) . from wall and readings averaged.

• Performance consistent with Manufacturer's specifications and intended application. Communicate tubing to exterior of chamber via sanitary adapter fitting and connect to particle counter. Measure every 10 minutes at flow rate of 1 CFM.Dry Heat Depyrogenation – OQ Particle counts: • Clean oven with IPA and lint-free wipes. • Sample from open end of 10 mm SS tubing placed at center of HEPA filter. .

Dry Heat Depyrogenation – OQ Chamber Pressure: • Measure pressure of chamber during cycle via sanitary port.C. relative to atmospheric as per Manufacturer's specifications. .03” W. • Cycle temperature set at ambient to avoid injury. • Differential pressure of > 0.

• Verify approved SOPs for operation and cleaning. • Verify maintenance and calibration schedule. • Assemble all vendor related documentation. • Verify spare parts entered into facility’s system. .Dry Heat Depyrogenation – OQ Confirm documentation: • Verify successful IO/OQ with signed report.

• Place thermocouples in chamber and within load to determine slow-to-heat zones within items being processed. cavities. . composition. complexity. • Calculate cycle parameters for desired process conditions and application. etc.Dry Heat Depyrogenation – HP Cycle Development/Heat Penetration Studies: • Prepare maximum load pattern. • Consider each item’s mass. Thermocouples to be placed in slow-toheat zones during subsequent PQ testing. density.

Dry Heat Depyrogenation – Endotoxin Challenge Endotoxin indicators: • Prepared from CSE. • Direct product inoculation preferred but may (will) present recovery issues. • Positive and Negative controls. • Usually prepared in 5 -10 sealed glass vials (ECV) placed within load at cold spots. .). • > 1000 EU (50 – 200% var.

Dry Heat Depyrogenation – Reduced Cycle • Perform at least one cycle under reduced cycle conditions (cycle setpoint reduced by 10 oC and exposure time reduced by 10 minutes) using a maximum load with thermocouples and endotoxin challenge indicators to demonstrate the robustness of the process (“worst case conditions”) and to accommodate potential process variations and variability in interpretation of process parameters. .

• Place endotoxin challenge vials adjacent to TC’s in load. .Dry Heat Depyrogenation – PQ • Place 12 TC’s in “worst case position” within load determined from HP studies. • Process 3 cycles. one TC at center of chamber. Place one TC at control probe.

36(5): 710-713. Technol.41 – 54 oC2). 36(1):23-27. T = temperature (actual temperature). J. Al.1982. . Tb = baseline temperature (250 oC). Z = z-value of endotoxin (46.Dry Heat Depyrogenation – PQ • Calculate FH at 250 oC: t (T – Tb)/Z FH = 0 ∫ 10 dt where: t = time at end of hold period. Environ. Mbio. 2Akers et. Parent. Appl. 1Tsuji and Harrison. 1978. Sci.

34 EU – (– 2 EU) = 5.Dry Heat Depyrogenation – PQ Calculate endotoxin log reduction value: • LRVendotoxin = log (A) – log (B) where: A = positive control B = test sample • LRVendotoxin = log (2200 EU/vial) – log (0.01 EU/vial) = 3.34 EU .

• Endotoxin positive controls > 1000 EU. • All endotoxin challenge exhibit > 3 log reduction. . • Endotoxin negative controls indicate no residual endotoxin contamination. • Control probe within + 5 oC of adjacent TC at midpoint of hold period. • Cumulative FH (250 C) values > 30 minutes.Dry Heat Depyrogenation – PQ PQ Acceptance criteria: • All TC’s within + 15 oC of setpoint (250 oC) during hold period.

• Review overall validation process and deviations to determine how process could be handled better in the future: – Carefully write protocols and acceptance criteria. – Ergonomics (maintenance/calibration). – Coordination with outside vendors. – Coordination with other ongoing activities to ensure required resources will be available when needed. try to anticipate problems or issues in advance. .Dry Heat Depyrogenation – Done! • All phases of validation successfully completed and final report signed off.

particularly with a new installation! .Dry Heat Depyrogenation – Done! • Allot extra time for validation. It always takes longer than you think.

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