Steam Sterilizer Validation Requirements Per The New Standard ISO 17665-1:2006

For decades, steam sterilization (autoclaving) has been an integral part in the manufacturing, cleanroom, and laboratory processes for the medical device, pharmaceutical, biologics, and human tissue/HCTP industries. It has been a common industry practice to validate steam sterilizers using the published guideline ISO 11134 Sterilization of health care products — Requirements for validation and routine control - Industrial moist heat sterilization,1 issued in 1994. In late 2006, AAMI released the document intended to supersede 11134, with ANSI/AAMI/ISO 17665-1:2006 Sterilization of health care products — Moist heat — Part 1: Requirements for the development, validation, and routine control of a sterilization process for medical devices.2 While other steam sterilizer guidance documents do exist,3,4 it is anticipated that the new 17665 standard will be recognized by the FDA and will be commonly employed to validate autoclave processes. The good news to manufacturers or other users of these guidelines is that many of the current validation practices are the same in the new document. This article will outline the basic requirements for steam sterilizer validation via the halfcycle overkill method, and list some of the differences between the two documents. REQUIREMENTS PRIOR TO VALIDATION The 17665 document makes it clear in numerous locations that the user’s quality system must adhere to ISO 13485:2003 Medical devices — Quality management system — Requirements for regulatory purposes.5 So if a user wishes to claim full compliance with the new 17665 steam standard, then their quality system must also be in compliance with ISO 13485, including items such as preventive/periodic maintenance and regular calibration for the sterilizer, documentation, change control, purchasing, etc. When compared with the previous steam document, the new 17665 also has more information on product and process characterization, sterilizing agent characterization, installation qualification/IQ, and operational qualification/OQ. The new document also states more clearly that a fully compliant validation is not just a series of successful halfcycles, but is the full complement of successful IQ, OQ, and PQ. Sterilization agent characterization will be simple for most users — moist heat/steam at 121 or 132 °C, and cycle selection (gravity, prevacuum, etc.). Process and equipment characterization means defining and documenting items like the sterilizer cycle parameters, products (or product families) to be sterilized, load configurations and limits, placement of biological indicators or chemical indicators (BIs/CIs), process tolerances, and equipment identification. Much of this type of information would be recorded in well-written validation protocols or validation final reports. Biological indicators often use spores of the bacterial species Geobacillus stearothermophilus at a titer of greater than 106per BI, although other species or titers are sometimes used. The new 17665 document also has more information on IQ and OQ. It defines IQ as “obtaining and documenting evidence that equipment has been provided and installed in accordance with its specification.” Autoclave installations commonly document items such as the sterilizer identification numbers, location, line voltage and amperage, water supply piping and pressure limits, steam line requirements, filtration, chamber size, structure and support, piping materials, software certification, manuals, drawings and documentation, and calibrations (temperature, pressure, and timer). The sterilizer must be installed in such a manner to facilitate any necessary maintenance, repair, adjustment, cleaning, and calibration.

OQ is defined as “obtaining and documenting evidence that the installed equipment operates within predetermined limits when used in accordance with its operational procedures.” Autoclave OQs commonly test or verify items such as cycle operation and programming instructions, safety and alarm testing, error reporting, empty chamber temperature profiling and chamber temperature limits/specifications, air removal testing, leak testing, temperature control anomalies, full cycle fullload temperature profiles (if proposed fullcycle exposure time is known), and determination of any hot or cold spots within the chamber. The product definition and process definition sections of the new document list things such as product specifications, product families, packaging, re-sterilization issues, package moisture, stability and potency of container products, re-usable container systems, process challenge devices/PCDs, sterility assurance level/SAL, BIs and CIs, and bioburden determination if necessary. PCDs are described as products or items that provide a known resistance to the sterilization process. They are commercially available or may also be created from the user’s product line by inserting spore strips, spore dots, inoculated threads, etc. into items or locations that are determined to be the most-difficult-to-sterilize product or location in the load. There are many other activities or decisions to be made prior to or during the IQ/OQ, that are not necessarily detailed in either standard. Items such as: ● Obtaining calibrated temperature recording devices or thermocouples ● Ordering supplies such as BIs, CIs, Bowie-Dick test packs, packaging materials, etc. and noting if adequate laboratory facilities are available ● Determining worst-case validation load and worst-case test product or PCD. The protocol or final report should contain a written rationale describing how the loads and product(s) were selected ● Selecting cycle type: 121 or 132 °C, gravity or prevacuum cycle, etc.; and determining if drying time needs to be qualified ● Is product bioburden testing necessary? ● Is product resterilization to be allowed and what are the requirements for resterilization? ● Is product stability or shelf life testing necessary for the user’s products? ● Does packaging testing or packaging validation need to be included with the protocol? VALIDATION – PERFORMANCE QUALIFICATION AAMI TIR #13 states “Sterilization process validation is a documented procedure for obtaining, recording, and interpreting the results required to establish that a process will consistently yield product complying with its predetermined specifications.” For the purposes of this article, the primary specification will be sterility. The performance qualification/PQ or microbiological qualification is a series of tests that establishes that the installed and properly operating sterilizer will process the users desired chamber loads to achieve the specified sterility assurance level/SAL. It must be remembered that the load is part of the validation — that is, if the user makes significant changes to the load at any point in the future — then re-validation may be necessary. The previous ISO 11134 document gave relatively little guidance information and few specifications for conducting the test cycles necessary to qualify the user’s proposed fullcycle exposure time(s). The

new 17665 steam document varies little from the previous standard in respect to the minimal PQ information that is provided. The 17665 describes bioburden validation methods and the more commonly used halfcycle “overkill” method. It should be noted that at the time this article was prepared, the proposed guidance document that is to accompany ISO 17665-1 was not yet available. This guidance document may provide more advice on microbiological qualification issues (ISO 17665-2 Sterilization of health care products — Moist heat —Part 2: Guidance on the application of ISO 17665-1). For this article, the general requirements for an overkill cycle PQ will be reviewed. While many activities are required to complete the PQ, the primary goal for the commonly employed overkill validation is this: the user needs to complete three consecutive successful halfcycles in order to qualify their proposed fullcycle exposure for routine processing of sterilization loads. In our case, successful means all BIs are killed (no growth upon incubation) for the three consecutive halfcycles. If, for example, there was no BI growth for the three test cycles at ten minutes exposure at 121 °C, then a 20-minute exposure at the same temperature would be adequate for routine daily processing, assuming all other aspects or requirements of the IQ/OQ/PQ are successful, documented, reviewed, and approved. But a description of the PQ needs much more detail than this. Validation protocols vary in format from company to company, but most will capture similar information for the final report. An example of validation protocol and final report sections would be: Title page with approval signatures • • Purpose, background information, or general goal(s) of validation Scope with more specifics about methods, cycles, facility, SAL, products and load, exclusions, etc. • References with published standards and company SOPs • Equipment, supplies, validation loads, BIs, etc. • Rationale for selection of products, load, cycles, PCDs, etc. • Procedure or methods (more details on this below) • Acceptance criteria which list the pass/fail requirements • Deviation report which lists any unexpected results, with potential effects on the validation, along with accept/reject rationale • Results and conclusions which assign a pass/fail decision to each acceptance criteria, summarize study, and include any requirements for revalidation • Attachment which lists any data sheets, diagrams, certificates, temperature records, etc., for inclusion with final report • Approvals section for final report. To conduct the halfcycles, the user assembles the worst-case validation test load, temperature loggers, BIs/PCDs, and CIs if necessary. The temperature loggers and BIs are seeded throughout the load to represent various chamber locations, keeping in mind any cold spots or previously determined most-difficult-to-sterilize locations. For small chambers, as few as five or six BIs and temperature loggers may be needed. Ten is a common sample size for many chambers. Large, multipallet-sized chambers may require many more samples per run. The sterilizer is programmed for one-half of the proposed full-cycle exposure time. Upon completion of the test cycle, the BIs are immediately removed and incubated, and the test load must be allowed to return to normal temperature prior to starting another test cycle. Temperature recorder data is downloaded and printed immediately to determine if any unusual temperature conditions existed. Information is entered on the data sheets (data sheets that would have been one of the attachments to the written protocol), and

The sterilizer must be added to a regular and documented calibration program. In summary. The completed final report packet must then be routed for review and signed for approval. or warnings. Including digital photographs of sterilizer. reviewed and signed. And the sterilizer must be added to the validation schedule for its annual requalification. including investigation and product quarantine procedure as appropriate. SOPs for daily processing must list all requirements for data that is to be reviewed and retained from the sterilizer runs. signed and reviewed.cemag. and load limit information must be readily available to all operators. 6) calibration documents for any measuring instruments used during the study. The sterilizer must be included in a regular and documented periodic/preventive maintenance program. filing system. . 3) notification of management or maintenance if sterilizer malfunctions or if recorder chart lists any errors. As stated before. Final reports should contain: 1) all sterilizer run data or recorder charts. conclusions. load. PCD.asp?pid=709 (5 of 7) [13/8/2008 11:56:50 AM] Controlled Environments® | Articles | Steam Sterilizer Validation Requirements Per The New Standard ISO 17665-1:2006 incubation period. growth promotion test cultures. The user needs to verify that all personnel that will be using the autoclave are trained using applicable operation and safety SOPs. loads. while leaving other areas (such as PQ) relatively unchanged. 4) any deviations recorded and investigated. all processed BIs must show no growth in order for the validation runs to be considered successful.all temperature records and data sheets are retained for the final report. etc. with logbook.us/Article_Print. SOPs must also address items such as 1) segregation of processed and non-processed product. 5) results. 2) storage requirements for processed products if necessary. Approved products. and 5) resterilization requirements if resteril-ization is to be allowed. BIs are checked regularly throughout the http://www. CI. POST-VALIDATION There are still issues to be addressed when all activities seem to have been completed. can be quite helpful for an auditor who may be reviewing the report at a later date. 2) all temperature recorder data. or archive for run records. there seem to be no drastic or revolutionary changes in making the transition from ISO 11134 to ISO 17665. signed and reviewed. and include positive control (unprocessed) BIs which must show growth. cautions. 7) the approved full-cycle parameters and acceptable placement locations for BIs for normal processing. The new 17665 steam document provides more information and more guidance in some areas. Untrained staff should not be allowed to run the sterilizer. 4) immediate notification of management for BI test failure. and discussion. or any other test results. and 8) manufacturers’ certificates of analysis for any items such as BIs. While users would be advised to obtain the 17665-2 guidance document when it becomes available. cycles. 3) all data sheets with BI. it is anticipated that manufacturers will not find any great difficulties in applying the new standard. growth media. etc. with final disposition.

Page 1 Validation of Steam Sterilizers Arden House 2009 James Gallagher Kalypsys Inc 1 Validation of Steam Sterilizers AAPS Arden House 2009 Page 2 Introduction to Validation of Steam Sterilizers Goals of Presentation • Overview of basic principles for steam sterilization and microbiology • Review key aspects of EN 285 and PDA Technical .pdf.com/meetings/files/139/James %20Gallagher. Google automatically generates html versions of documents as we crawl the web.aapspharmaceutica.This is the html version of the file http://www.

Report #1 • Facilitate decision making for Pharmaceutical Scientists looking to Contract Manufacturing Organization (CMO) for aseptic processing that includes steam sterilization 2 Validation of Steam Sterilizers AAPS Arden House 2009 Page 3 Introduction Outline • Introduction • Sterilizer Design Aspects • Steam / Thermo • Micro Aspects • Validating Sterilization Cycles • Checklist / Troubleshooting .

March 2008 USP <55> Biological Indicators. Resistance Performance Tests PDA Technical Report #1: Validation of Moist Heat Sterilization Processes: Cycle Design. purity and control EN285 Sterilization – Steam Sterilizers – Large Sterilizers. Amendment 1. Development and Ongoing Control Validation of Steam Sterilizers AAPS Arden House 2009 . Guidance for Industry Sterile Drug Products Produced by Aseptic Processing — Current Good Manufacturing Practice . Sept 2004 CFR Relevant Sections on Sterilization.• Regulatory 3 Validation of Steam Sterilizers AAPS Arden House 2009 Page 4 Introduction to Validation of Steam Sterilizers Regulatory Drivers and Guidance Source Document FDA FDA.

and the RFP has taken into account the level of validation needed • Review the assumptions and risk analysis on the required steam sterilization processes are included.4 Page 5 Introduction to Validation of Steam Sterilizers Working with Contract Manufacturing Organizations (CMO) • Sponsor needs to partner with CMO • Assume aseptic process is defined. confirm assumptions in original risk assessment are still valid • Consider including an engineering (pilot) batch in the RFP .

• Due Diligence: site visit.Leverage CMO’s existing cycles / programs WW Phase III Clinical Program . audit → Input on CMO selection • Transferring Aseptic Process Technology to the CMO 5 Validation of Steam Sterilizers AAPS Arden House 2009 Page 6 Introduction to Validation of Steam Sterilizers Three Examples of Contract Services Validation of Steam Sterilizers AAPS Arden House 2009 6 Example of Project Typical Sterilization Needs Driving Forces Early Phase Clinical Program in US Receiving Tank / Bag Documentation for IND Speed to Clinic -.

Filter Assembly Filling Line Parts Documentation to support NDA/ CTD and PAI Scale-up Secondary source for Commercial Product Stopper processing Filling Manifold Supplement to NDA / CTD New facility Qualification Equivalency with existing product / package/ process Optimal cycles Page 7 Sterilizer Design Aspects Sterilizer interface with aseptic process • Sterilizers are a critical means to provide access to an aseptic process • Components are prepped /cleaned .

prior to sterilization • Dedicated. insulation . steam trap. disposable or multiple use Validation of Steam Sterilizers AAPS Arden House 2009 7 Prep Area (non sterile) Entry Sterilizer #1 Sterilizer #2 Sterilizer #3 Aseptic Processing Suite Aseptic Corridor Page 8 Sterilizer Design Aspects Autoclave Diagram Validation of Steam Sterilizers AAPS Arden House 2009 8 Page 9 Sterilizer Design Aspects Sterilizer Design Features • Jacketed vessels. internal volume.

standard door interlocks • Condensate trap within 2 meters of the connection • May have air breaks on drains to prevent backflow Validation of Steam Sterilizers AAPS Arden House 2009 9 Page 10 Sterilizer Design Aspects Control of a sterilizer • Critical parameters are steam quality.• Filtered air (< 0.3um) with steam backflow device • Temperature sensors = Pt resistance types • 2 independent temperature sensors • Failed cycle can be vented and loading door opened. temperature and time .

Drain. pressure and time 10 Validation of Steam Sterilizers AAPS Arden House 2009 Common Indicators on Sterilizer Controller Door lock (both ends) Cycle in progress / Cycle Complete Fault Cycle selected Cycle counter Cycle Stage Indicator Page 11 Sterilizer Design Aspects Temp and Pressure Measurements on Sterilizer • Temperature Measurements – Jacket.• Temperature sensor is normally in the drain • Automated cycles. sensors monitoring and alarms • Key measurements are temperature. Load Probes and Recorders • Pressure Measurements . Chamber.

Chamber and Recorders • Time – Exposure time.– Jacket.01 bar) Page 12 Steam / Thermo Steam Overview– why is steam so effective? • Condenses.6% over range 0 to 400kPa / (-1 to 3 bar) Accuracy at Sterilization 0.05 bar) Resolution 0. heat up time Validation of Steam Sterilizers AAPS Arden House 2009 11 Parameter Temperature Pressure Accuracy of Range 1% over the 50 – 150˚C range < 1.1˚C for digital 1kPa (0.5˚C at sterilization temp +/-5 kPa (0. collapses as wet film of condensation increases the heat flow to sterilized item .

• Volume of steam ( ~6 cu ft / lb). 350x volume of water • 50 – 100 lbs of steam used in a typical cycle • The “killing power” of steam is due to its latent heat of vaporization – 1 L water to boiling = 80 cal – 1 L boiling water to steam = 540 cal 12 Validation of Steam Sterilizers AAPS Arden House 2009 Page 13 Steam / Thermo Steam Enthalpy 13 Validation of Steam Sterilizers AAPS Arden House 2009 Temperature (°C ) Pressure (bar) (psig) Enthalpy of Steam (J/g) (BTU/lb) .

013 14.725 1.715 1.707 1.0 for dry saturated steam.1 2. less .392 34.167 134°C 3.150 121° C 2.675 1.039 44.7 2.164 126˚C 2.048 29.7 2.7 2.171 Page 14 Steam / Thermo Steam Quality / Testing • Steam quality = how much water is contained in the steam % by weight / % by volume • Dryness value = 1.100˚ C 1.

minimal NCG • Clean steam used in Pharma applications. condensate complies with WFI monograph • Understand CMO’s limitations for steam production 14 Validation of Steam Sterilizers AAPS Arden House 2009 Page 15 Steam / Thermo Causes of poor steam quality • Issues with clean steam generator • Water hammer – water slug moving through pipes resulting in a banging sound .latent heat capacity for lower steam quality • Pharma sterilization cycles use saturated steam with no superheat.

startup • Condensate in piping: in AM. steam/gas mix .• Piping Insulation– prevents steam from condensing • Times of higher steam demand: winter. move to an area of lower velocity (the sterilizer). after a shutdown Validation of Steam Sterilizers AAPS Arden House 2009 15 Page 16 Steam / Thermo Non-condensable gases(NCG) • Gases that cannot be liquefied by compression under the conditions used in a sterilization cycle • NCG do not contract / expand like steam.

10% air will lower incoming steam temp by 7°F • CO 2 can dissolve in the condensate → carbonic acid. but traps steam in the system.• Sources: Air: open door. piping.e. vents air. can insulate items → impact cycles – i. located on drain legs and . steam supply • Lower temperature. corrosive to metal pipes • Oxidation from dissolved 0 2 Validation of Steam Sterilizers AAPS Arden House 2009 16 Page 17 Steam / Thermo Steam Trap • Automatic valve that drains water.

z and F values Microbiological Aspects of Sterilization . F0 Biological Indicators Overkill or Bioburden Cycle? D.steam filters • Located at the bottom of the sterilizer. drains condensate from the jacket and the chamber • Steam traps also used on air vents • Failure mechanisms for steam traps Validation of Steam Sterilizers AAPS Arden House 2009 17 Page 18 Micro Aspects Overview Validation of Steam Sterilizers AAPS Arden House 2009 18 SAL = 10-6 Accumulated Lethality.

often called a six log reduction • PNSU (Probability of a Non-Sterile Unit) • Cannot directly measured this objective • For parenteral products.Page 19 Micro Aspects Sterility Assurance Level (SAL) • The probability of a single viable microorganism being present on a sterilized unit is one in one million after the item has undergone a sterilization process. desire a SAL of 10-6 19 Validation of Steam Sterilizers AAPS Arden House 2009 Page 20 Micro Aspects Biological Indicators (BI) .

• Population of microorganisms (usually spores) inoculated onto a suitable medium • Placed in sterilizer load locations to determine the sterilization cycle efficacy by deactivating BI • The challenge microorganism is selected based upon its resistance to the given process • Quality of BI defined by microbiological count and D-Value Validation of Steam Sterilizers AAPS Arden House 2009 20 Page 21 Micro Aspects G Stearothermophilus • Geobacillus stearothermophilus .

2 min • Desire a population of spores on a strip of ~105 or more • Direct inoculation onto test substrates (closures etc) Validation of Steam Sterilizers AAPS Arden House 2009 21 Page 22 Micro Aspects Inoculation of BI / Positive Controls . commercial spore strips have D-value 1. highly resistant to heat • Most spore forming microbes have Dvalue < 0.1°C to 135°C (275°F) • Incubate at 55˚.60°C • Thermophiles found in hot springs areas such as Yellowstone NP.for use in steam sterilization at 121.5 .5 min.

• After the sterilization cycle. no change in indicates sterilization conditions were achieved. careful consideration should be given during sterilization validation to the nature or type of material chosen as the carrier of the biological indicator to ensure an appropriately representative study” Sept 2004 FDA GUIDANCE. “It also should be noted that the resistance of microorganisms can vary widely depending on the material to be sterilized. the retrieved BI is placed in a tube of growth medium and incubated per USP <55> • A color and/or turbidity change indicates the results of the sterilization process. For this reason. . otherwise the growth of the spores indicates that the sterilization process has not been met.

• Typical D-values for commercial spore strip lots are ~1. or 90% reduction in population.Validation of Steam Sterilizers AAPS Arden House 2009 22 Page 23 Micro Aspects D-value • D value is the thermal resistance value (min) of a target organism • D value is the time in minutes at a specific temperature to reduce the surviving microbial population by 1-log.5 .2 min Validation of Steam Sterilizers AAPS Arden House 2009 23 Page 24 Micro Aspects .

Z Value • Z value is the heat resistance of a spore as a function of temperature (°C) • Z value is the temperature change required to result in a 1-log reduction in D-value • Generally used standard value is Z= 10˚C • Z = (T 2 –T 1 ) / (log D 1 – log D 2 ) Thermal Resistance Curve Validation of Steam Sterilizers AAPS Arden House 2009 24 D 121 1.6 min D 131 .

1)/z x t Where T = Temperature F 0 = equivalent sterilization time (min) • Z = 10˚C is generally used Validation of Steam Sterilizers AAPS Arden House 2009 . F = Σ 10(T-121. Accumulated Lethality • Accumulated Lethality is the F value •F 0 is the equivalent time that a microbial population with a z value of 10 has been held at 121˚C •1F 0 = the equivalent of 1 minute at 121˚C • Equation.0.16 min D 111 16 min Page 25 Micro Aspects F value.

2 min. natural is <0. D and Z values and their relationships • Equation #1: Log N F = -F (T.5 .25 Page 26 Micro Aspects F.z) = (Log N 0 – Log N F )xD T • Typical D-values are ~1.5 min .z) /D T + log N 0 • Equation #2: F (T.

z) = (Log N 0 – Log N F )xD T .26 Validation of Steam Sterilizers AAPS Arden House 2009 Page 27 Micro Aspects Product Specific (Bioburden) Approach • Quality attributes impacted by high thermal input • Collect detailed bioburden and D-value data • Example: Liquid Loads. terminal sterilization with a bioburden of 100 CFU and D value = 0.5 min • Equation #2: F (T.

0 min at 121˚C 27 Validation of Steam Sterilizers AAPS Arden House 2009 Page 28 Micro Aspects Overkill Approach • Many definitions and process requirements for Overkill Cycles • Provides a minimum 12 log reduction of microorganisms having a D-value of at least 1 min • Avoids collecting bioburden and Dvalue data by assuming extreme case conditions: – Bioburden level is 106 – D-value is 2.5 minutes .–F 0 = (Log 102 – Log 10-6 ) x 0.5 min = 4.

1˚C 28 Validation of Steam Sterilizers AAPS Arden House 2009 Page 29 Micro Aspects Compare F PHY and F BIO •F PHY determined from thermocouple data during heat .5 min/log) = 30 minutes at 121.• Equation #2: F (T.z) = (Log N 0 – Log N F )xD T F 0 = 12 log (2.

penetration study •F BIO is the delivered lethality calculated by the actual kill of microorganisms in a BI system •F BIO =D T x LR D is the D value LR is the log reduction of BI population during a cycle • Agreement between F PHY and F BIO Validation of Steam Sterilizers AAPS Arden House 2009 29 Page 30 Micro Aspects BIER vessel .

Sept 2004 30 Validation of Steam Sterilizers AAPS Arden House 2009 Page 31 . purity • Confirms D-values from commercial lots of spore strips – Fractional Negative or Direct enumeration methods – Repeat value or confirm by survivor kill “The microbial count of a biological indicator should be confirmed. Biological indicators should be stored under appropriate conditions.” FDA.• BIER (Biological Indicator Evaluator Resistometer) Systems are designed to provide environmental conditions to evaluate the resistance of microbial populations to sterilization • Confirm population of spores on strip.

Validating Sterilization Cycles Overview Validation of Steam Sterilizers AAPS Arden House 2009 31 Thermal Validation System Type of Goods Worst Case Load Assessment Load Configuration Thermocouples Overkill or Bioburden? Page 32 Validating Sterilization Cycles Aspects of Thermal Validation System • System that meets international cGMP requirements .

• Performs pre and post calibration of thermocouples • Consider IRTD for reduced setup time. minimal sensor handling and automated sensor calibration • High Temperature range -195 to 420°C • 21 CFR Part 11 Compliant • Calibration Traceable to NIST • Facilitates study data and generates regulatory required reports 32 Validation of Steam Sterilizers AAPS Arden House 2009 Page 33 Validating Sterilization Cycles Thermocouples • Number must be documented and justified. watch acceptance criteria! .

5˚C • Type T Class 1 thermocouple wire • Use of telemetry sensors • TCs threaded through gland to the chamber • Pre and post calibration verification “The sensing devices used for validation studies should be calibrated before and after validation runs” FDA Sept 2004 Guidance”.0.= Red Wire material: + = Copper .= Constantan Properties: . 33 Validation of Steam Sterilizers AAPS Arden House 2009 Page 34 Validating Sterilization Cycles Type T Thermocouples • Type T Thermocouple • Wire insulation color: + = Blue .• Accuracy of thermocouples = +/.

+ = Copper color Validation of Steam Sterilizers AAPS Arden House 2009 34 Junction Exposed Un-Grounded Grounded Tip Page 35 Conducting Validation More on Thermocouples 35 Validation of Steam Sterilizers AAPS Arden House 2009 • Number / access may result in failed test cycle • Can be easily damaged by autoclave cart wheels • Essential that the thermocouples do not affect air removal or steam penetration • Label thermocouples • Conducting verification regularly (not necessarily .

Gowns. Vessels with vents . the biological indicator should be placed adjacent to the temperature sensor so as to assess the correlation between microbial lethality and predicted lethality based on thermal input” FDA Sept 2004 Guidance. Filters.after each run) • Place one near the sterilizer’s temperature sensor “In general. Piping. Glassware 1 or more (3 typically) Easy air removal and steam penetration Wrapped Goods Hoses. Page 36 Validating Sterilization Cycles 3 Classes of Goods to be sterilized Type Goods Sterilized Typical PreVac Air Removal Hard Goods Equipment.

3 or more More difficult air removal/ steam penetration Liquids Media. 37 Validation of Steam Sterilizers AAPS Arden House 2009 . as well as biological indicator and temperature sensor locations. Batch production records should subsequently document adherence to the validated load patterns” Sept 2004 guidelines. Heat and cool without vacuum 36 Validation of Steam Sterilizers AAPS Arden House 2009 Page 37 Validating Sterilization Cycles Establishing Load Patterns “The specific load configurations. should be documented in validation records. Product (F 0 > 15 min) None Air overpressure process.

Load Position Comment Fixed Fixed Identical for all processing runs Instructions list items and position in diagram Fixed Variable Location can vary Validate positional equivalency during runs Instructions reference list of items Variable Variable Location and position can vary Validate minimum and maximum loads Demonstrate min / max are adequate in validation Flexible instructions for Operations Page 38 Validating Sterilization Cycles Types of Saturated Steam Processes • Pre-vacuum process is most commonly used saturated steam process – Removes air in vacuum pulses .

– Multiple pulses allow pre-conditioning of goods. reducing the equilibration time • Gravity Displacement – Steam displaces the heavier air – Air pushed out drain through steam trap – Steam distribution is critical Validation of Steam Sterilizers AAPS Arden House 2009 38 Page 39 Validating Sterilization Cycles Cycle development for new item • Establishing a cycle prior to validation • Assessment of the item / current cycle adequate? – Class of goods. OK or bio? • Conduct heat penetration study • Determine equilibration time: time T REF . type of load.

seal – Bottles – center. insert. just above bottom – Filters – air removal .– time T slow • Drying studies if needed • Correlate F PHY and F BIO for cycle Validation of Steam Sterilizers AAPS Arden House 2009 39 Page 40 Validating Sterilization Cycles Determination of Worst Case Load • Determine locations that are worst case with steam integrators and/or Thermocouples • Determine most difficult to sterilize items in load – Hoses – cut.

• Fixed load or variable load -.degree of flexibility desired in CMO’s Operations • May be a destructive test . pass through expense Validation of Steam Sterilizers AAPS Arden House 2009 40 Page 41 Validation of Steam Sterilizers AAPS Arden House 2009 41 Validating Sterilization Cycles Definition of Loads – Contract Service Examples Contract Service Goods Type Load OK or Bio? Cycle #1 Vessel Wrapped Fixed Overkill 121˚C saturated steam #2 Filter Wrapped .

3mbar/min Check prior to thermal studies Monitors rise in pressure under vacuum .Fixed Overkill 121˚C saturated steam #3 RTS Closure Wrapped Variable Overkill 121˚C saturated steam Page 42 Validating Sterilization Cycles Example Cycle Overview Validation of Steam Sterilizers AAPS Arden House 2009 42 Page 43 Validating Sterilization Cycles Types of Studies Cycle Description Standard Key Aspects Pressure Rise (Leak Test) < 1.

probes to set point Item Cool Point Min load Multiple runs if needed Identifies the most difficult to sterilize point in a test article Validation of Steam Sterilizers AAPS Arden House 2009 43 “These uniformity or mapping studies should be conducted with calibrated measurement devices. probe to probe. probes to set point .” FDA Sept 2004 Guidance Page 44 Validating Sterilization Cycles Types of Studies Cycle Description Standard Key Aspects Temperature Distribution Min / Max Load Verifies uniform distribution of steam Temperature variation: each probe. probe to probe.Temperature Distribution Empty Chamber Often performed in requal programs Verifies uniform distribution of steam Temperature variation: each probe.

Heat Penetration Max Loads multiple runs Map temperature with TCs Temps as above Equivalency of variable loads using same cycle Maximum equilibration time Used to calculate F PHY Can be combined with BI challenge study Process Lethality Biological Qualification (BI challenge) Full Load 3 consecutive runs Map temperature with TCs Place BI at probed locations. including most difficult to heat Used to calculate F BIO Incubate BI post cycle 44 Validation of Steam Sterilizers AAPS Arden House 2009 Page 45 Validating Sterilization Cycles Minimum Acceptable Cycle .

• Qualified MAC cycles confirmed biologically and physically • Safety margin through use of higher exposure times or temperatures • Total Dwell Time is additional lethality + demonstrated lethality from process validation • Half cycle methods Validation of Steam Sterilizers AAPS Arden House 2009 45 Page 46 Validating Sterilization Cycles Acceptance Criteria Guide • Thermal Systems • Process Cycles .

• Reference Tests • EN285 Steam quality items Validation of Steam Sterilizers AAPS Arden House 2009 46 Page 47 Validating Sterilization Cycles Acceptance Criteria–Thermal Systems Aspect Standard TC Temperatures during Dwell Time All temps during dwell time within 3˚C (-1˚C /+2˚C) of SP TC Temperatures during Dwell Time Fluctuation of TCs within chamber NMT 1˚C TC Temperatures during Dwell Time All temps measured in chamber do not differ from each other by 2˚C Steam Temperature Corresponds to its vapor pressure measurement Equilibration Time Lag between hottest / coldest thermocouples is NMT 30 sec (15 sec for smaller chamber) Timer Accuracy .

length of hoses F 0 Range .1% Pre and post calibration check Temp measurement system is accurate to +/-0. large mass.+/.5˚C Validation of Steam Sterilizers AAPS Arden House 2009 47 Page 48 Validating Sterilization Cycles Acceptance Criteria– Process Cycles Validation of Steam Sterilizers AAPS Arden House 2009 48 Aspect Standard Concerns Temperature Distribution Minimum F 0 met for TCs Correlate T and P Acceptable number of TCs Heat Penetration Determine most difficult to sterilize item / desired load Air removal.

Min F
0

at end of exposure Max equilibration time Microbial Inactivation during BI challenge No BI growth Growth with + control SAL = 10-6 Positive control of BI SOPs for handling BI Comparison of F
PHY

and F
BIO

Agreement for minimal cycle
Page 49

Validating Sterilization Cycles Acceptance Criteria – Reference Tests
Aspect Standard Load Dryness Mass increase < 1% Textiles test pack Mass increase < 0.2% Metal test pack Dynamic Pressure Test Average pressure change NMT 10 bar/min for any 3

second interval Sound Power Sound level meter reading NMT 3dB change from original operating level
Validation of Steam Sterilizers AAPS Arden House 2009 49

• Reference tests performed during validation of cycle, revalidation and in periodic / routine tests • Most test are done on an empty chamber
Page 50

Validating Sterilization Cycles Acceptance Criteria – Reference Tests II
Aspect Standard Thermometric Tests (Full Load) Equilibration time NMT 30 sec (large) TCs within +3°C of sterilization temp Minimum hold time at sterilization temp Temps within 2°C during hold time

Chamber steam temp corresponds to pressure Bowie and Dick Test Uniform change of indicator color Air Leakage Flow Rate NMT 1.3mbar/min Air Detector (if present) Alarms if <2°C temperature difference at start of equilibration time Hollow Load Test Process challenge device reaches endpoint
Validation of Steam Sterilizers AAPS Arden House 2009 50 Page 51

Validating Sterilization Cycles Acceptance Criteria -- EN285 Steam Quality
Item Description Limits Non condensable gases (NCG) Air and other gases, which do not condense under the conditions of steam sterilization and prevent the attainment of sterilization conditions in any part of the load < 3.5% Superheat

at any given pressure.95 Contaminants Clean steam condensate tests per EP EN285 Table E. the absence of air leaks and steam penetration into a porous load • Test Sheet consists of chemical indicators on two test .2 Validation of Steam Sterilizers AAPS Arden House 2009 51 Page 52 Validating Sterilization Cycles Bowie-Dick Test / Steam Penetration • Test is designed to test air removal. is higher than that indicated by the equilibration curve for the vaporization of water ≤ 25°C Dryness Value The dryness fraction is a measure of the amount of moisture carried by the steam being supplied and used for sterilization 0.90 – 0.Steam whose temperature.

Retention of air within the pack due to: – adequacy of pre-vacuum – air leak – presence of NCG in steam supply • Bowie –Dick cycles: to be carried out periodically per EN285 .sheets positioned inside porous materials and sealed inside a disposable outer wrap • Single use packs available 52 Validation of Steam Sterilizers AAPS Arden House 2009 Page 53 Validation Sterilization Cycles More on Bowie and Dick Test • Successful test indicates rapid and even penetration of steam into the test pack.

end of cycle adjustments as needed with post vacs. heating . – Initial qualification – demonstration of equivalency – bracketing approach – supported by risk assessment • Ongoing monitoring of cycles • PQ studies + operational efficiencies if equivalent. Otherwise pick coolest one • Drying studies.• Test pack can also be used for load dryness test (textiles) Validation of Steam Sterilizers AAPS Arden House 2009 53 Page 54 Validating Sterilization Cycles Assessing Limitations / Restrictions • Qualification of multiple autoclaves.

Wrapped Good – 121˚C sat steam cycle with pre-vac – Fixed Load Configuration (min load) – Thermal mapping Study – Heat Penetration Study / BI Challenge Study • Include documentation for sterilization process in IND Validation of Steam Sterilizers AAPS Arden House 2009 .54 Validation of Steam Sterilizers AAPS Arden House 2009 Page 55 Validating Sterilization Cycles Contract Service Example #1: Post Sterile Filtration Vessel (Tank / Bag) • Assumes an early phase clinical program in US • Will an existing cycle provide sufficient assurance that the new load will achieve SAL? Bracketing strategy? – Overkill Cycle.

wrapped good – 121˚C sat steam cycle with pre-vac – Fixed load configuration – Thermal mapping study – Heat Penetration – BI Challenge • Include documentation for sterilization process in CTD Validation of Steam Sterilizers AAPS Arden House 2009 56 Page 57 .55 Page 56 Validating Sterilization Cycles Contract Services Example #2: Filter Assembly • Assumes a late phase WW clinical program / CTD filing • Steam sterilizer cycles will be held to EN285 standard for Regulatory filings and PAI inspections – Overkill Cycle.

but may consider bioburden based cycle – Variable load. wrapped good – Thermal mapping study – Heat Penetration – BI Challenge: direct inoculation on closures – Drying studies may be needed .Validating Sterilization Cycles Contract Service Example #3: RTS Closures Validation of Steam Sterilizers AAPS Arden House 2009 57 • Assumes sponsor is partnering with CMO to source existing product. closures cycle not currently qualified • WW sourcing --Steam sterilizer cycles will be held to EN285 standard for Regulatory filings and PAI inspections – OK.

cleaning sterilization and storage of specified equipment and materials – Confirming cycle parameters are listed on Master Production Records .• Include documentation for sterilization process in CTD Page 58 Checklist Sponsor’s Checklist of CMO supplied documents • Explanation and justification of method of sterilization – Diagram showing location of load items. printouts – Summary of micro results – Pre and post calibration of thermocouples • Information on preparation. BI locations. TC locations.

impact to utility system changes assessed? Type. Cycle Description Form (CDF) for load configuration changes covered. plant environmental isolates How is micro data reviewed and approved? Requalification program in place NIST traceable standards used.– Material and Personnel flow diagrams Validation of Steam Sterilizers AAPS Arden House 2009 58 Page 59 Checklist Quality and Micro QUALITY AND MICRO CHECKLIST Change control System. D-value. review how sensors are calibrated Compare process record. spec for time and temperature requirements Control Software and documentation should be fully traceable through the project with all documentation accurately reflecting the changes and developments made throughout the project lifecycle Filing Documentation : Reports. MBR for indicated product. MBR with documents for the sterilization . source. Z. concentration. SOPs.

and warning alarms Review PM schedule. temp. control system. alarms. cold spot monitoring.process Experience of key personnel and staff Validation of Steam Sterilizers AAPS Arden House 2009 59 Page 60 Checklist Sterilizer Design STERILIZER DESIGN ITEMS Understanding of sterilizer control system and parameters that will be included on batch documentation Material Flow / Building: floor plan. internal volume. any limitations identified Integrity testing and sterilization of filters for vacuum break Validation of Steam Sterilizers . placement of autoclaves and critical items Utilities: clean steam. jacket pressure. materials of construction. filters used. location of controller sensor. compressed gasses free of particulates and oil vapor Mfg of sterilizer. How often is steam trap checked Adequate generation and distribution of Clean steam.

hold times. cold spot for each pattern Min / Max load configurations: hi/low avg Temp during dwell. ID Time limits established. compare load patterns with SOP and cycle forms. cold spot. run date/time. allowable variation Validation of Steam Sterilizers AAPS Arden House 2009 . sampling instructions. min/ max F0. dwell time. results for positive controls + tested BI Heat penetration cycles: load pattern. conformance to specifications Sterilization process description: Validation info for BI. heat penetration. periodic leak tests Program to check / monitor Steam Quality. ensure that saturated steam is used Methods and controls to monitor routine cycles Any adjustment to cycles for equilibration times Empty chamber cycles: # runs. demonstrate uniformity reproducibility. # of runs.AAPS Arden House 2009 60 Page 61 Checklist Sterilization Cycle Items STERILIZATION CYCLE ITEMS Protocols and data summaries that justify steam cycle. loading patterns.

PQ. MBRs on pivotal batches Gap Analysis or Documentation that sterilizer is capable of meeting EN285 List of registrations and licenses Site inspection history Prompt delivery of reports and data Confirm that sterile prep area has documented SOPs Procedural controls: SOP or Cycle sheets to include: load pattern.61 Page 62 Checklist Business and SOP Checklist BUSINESS AND SOPS CHECKLIST Documentation to support Regulatory Filings: supplied in reports or referenced in a DMF with authorization letters. IQ. training. OQ. logbooks Validation SOP. Micro programs Number and distribution of thermal monitors listed in SOP Prep Team for Pre-Approval Inspection Training: SOPs and Operator training records Validation of Steam Sterilizers AAPS Arden House 2009 62 Page 63 . revalidation.

The agency says the firm has not verified and validated its production . “These deviations raise significant concerns with sterility assurance of products that were produced under these conditions.” said the agency Connecticut-based _______ for its sterile manufacturing practices by the FDA.Regulatory Some FDA Observations to Sponsors Validation of Steam Sterilizers AAPS Arden House 2009 63 According to FDA documents. the firm also was written up because equipment and supplies used to work on. were not kept separate from the supplies used in product manufacturing as necessary to prevent cross contamination" San Diego-based _____________ has been warned by the FDA for not adhering to procedures to prevent microbial contamination of sterile pharmaceuticals. or exposed to pathogenic and potentially pathogenic agents.

Guidance for Industry: Submission Documentation for Sterilization Process Validation in Applications for Human and Veterinary Drug Products. • Validation of Moist Heat Sterilization Processes: Cycle Design. Practical Guide to Autoclave Validation. Qualification and Ongoing Control. • Agalloco. The violations “may be symptomatic of serious problems in your firm's manufacturing and quality assurance systems. Amendment 1. Raymond G. James. Sept 2004 • FDA. including sterilization and packaging of devices. July/ Aug 2002 • EN285:2006+A1. March 2008 • FDA. Vol 61. Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing — Current Good Manufacturing Practice.” according to the letter. Technical Report No. Development.processes. Sterilization – Steam Sterilizers – Large Sterilizers. PDA Journal of Pharmaceutical Science and Technology. Pharmaceutical Technology . Pharmaceutical Engineering. Understanding Overkill Sterilization. No S-1 • Lewis. 1 Revised 2007.