USP <1116> and its impact on Microbiology Dr. Tim Sandle Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Introduction EM guidance Background to USP <1116> Main changes and debates Method limitations Incident rates Frequencies of monitoring Locations of monitoring Other changes Regulatory issues Rapid methods Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Sources of EM Guidance EU GMP - last EM revision 2009; new annex 1 2016? FDA aseptic filling guide (2004) PDA Technical Report (2014, 3rd revision) ISO 14698 (1998). Future update? USP <1116> (2011, published 2012) Pharmig Current Review (2010) PHSS Biocontamination control guide 2015 Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Sources of EM Guidance ISO 14698 Under long-term review Possible development to a viable cleanroom classification standard (like ISO 14644) No other standard is likely to be reviewed in the short-term Pharmaceutical Microbiology: http://www.pharmamicroresources.com

ISO 14644 ISO 14644 Parts 1 and 2 revised in December 2015 A standard for cleanroom classification. Some detail about on-going monitoring. Does not cover viable monitoring at all. Changes: New look-up tables Increase in counter locations Changes to sampling volumes Each individual location must pass. More risk based. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP <1116> (2012) USP <1116> Revision began in 2005 Objectives of USP committee: Focus the chapter on environmental monitoring only, removing information relating to aseptic process validation. Focus the document exclusively on the monitoring of aseptic environments. Reconsider the alert and action level (limit) concept. Effective 1st May 2012, 35th edition of the USP Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes Former title: “Microbial Control and Monitoring Environments Used for the Manufacture of Healthcare Products” Revised title: “Microbiological control and monitoring of aseptic processing environments” Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes Scope: Environments Pharmaceutical sterile products Bulk sterile drug substances Sterile intermediates Excipients Environments Conventional clearoom with UDAF Blow-fill-seal RABS Isolator Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes The emphasis on the word “aseptic” in the introduction implies that the chapter is not applicable to all “sterile” products. This means that terminally sterilised products are outside the scope of the chapter. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes By “aseptic” a low level of contamination is acknowledged: “an expectation of zero contamination at all locations during every aseptic processing operation is technically not possible and thus is unrealistic” Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes USP notation system dropped: M3.5 etc. And old FDA 209E classes e.g. Class 100, Class 10,000) Replaced by ISO 14644 classes in the operational state Difference with EU GMP: Class 5 = EU GMP Grade A Class 7 = EU GMP Grade B Class 8 = EU GMP Grade C Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main Changes Relative risks Picture showing on how process separation and product protection interact (adopted from Bioquell U.K Ltd). Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main changes The design and construction of clean rooms and controlled environments are covered in ISO 14644. ISO 14644 stipulates the total particulate counts required for a clean environment to meet the defined air quality classifications. USP accepts this standard verbatim. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Main Changes New guidance for cleanroom operations: ISO class 8 = 20 air changes per hours ISO class 7 = 50 air changes per hour ISO class 5 = 100 air changes per hour Isolators = can have a different justification for air changes and air velocity An EM programme should only be constructed and executed once airflow mapping and HVAC dynamics have been optimised. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Environmental monitoring USP places emphasis upon particle and viable monitoring Particle counting most important for isolators Viable monitoring is regarded as semi-quantitative Trending is the most important aspect of the monitoring programme Isolated counts “a normal phenomenon in conventional cleanrooms” which do not require specific corrective action and there is the possibility of a false positive Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Environmental monitoring USP requires monitoring of: Surfaces Air (room and enclosure) Compressed gas Changes in trend must be investigated, and include assessment of: Maintenance Disinfection Unusual events and activities Physical changes e.g. temperature and humidity Staff training Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Environmental monitoring SCDM (TSA) is a suitable medium, incubation at ‘low’ and ‘high’ temperatures (20-35oC for not less than 72 hours) Consideration given to fungal medium e.g. SDA Certain conditions may require micro-aerophilic monitoring e.g., certain gasses or sterility test failure using anaerobic media Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Methods Settle plates: EU GMP: Semi-quantitative measurement (CFU / 4hours) Require desiccation study. USP 1116: “The exposure of open agar-filled Petri dishes, or settling plates, is not to be used for quantitative estimations of the microbial contamination levels of critical environments.” Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Monitoring methods No EM programme can prove sterility Environmental control is the most important, supported by EM and media simulations EM can demonstrate that a clean room is operating with in a consistent state of control However, “EM” requirements have evolved in a manner that did not fully consider analytical capability and metrology Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Monitoring methods All monitoring methods are flawed: Require people to take them Variability in technique Risk of false positives No one method can detect all types of contamination All methods have relatively poor recoveries (“insensitive”) Air-samples are particularly weak Surface methods have poor recoveries Settle plates are not considered quantitative All methods are poor at recovering damaged or stressed microorganisms Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Monitoring methods Therefore, numerical targets (as CFU) should: Not be used as limits (they are “levels”) Not be considered specifications Be seen as informational only Low or zero counts are not, by themselves, guarantees for microbial control; Equally, excursions beyond numerical limits are not necessarily and indication of loss of control. Instead: Count non-zero events. Use a contamination recovery rate metric based on historical findings Inference that these should be stable over time with little variation. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Plate counting debate DEBATE Should not simply count CFU due to the inaccuracies of current environmental monitoring methods It is more important to count the incidence rates and investigate out of trend. The limit of quantitation (the number of cfu that can be reported accurately) is 15. If CFU below 15 cfu , do not worry if trend is OK. If above 15 cfu, investigate. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

What does this mean? At very low recovery levels there is no way to establish Alert or Action Levels statistically-the counts are simply too low to make statistical analysis useful. Instead, emphasis should be on incidents. “Hits” in ISO 5 aseptic environments should be infrequent. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

What was the pre-USP 2012 situation? Take active air-samples: Microbiological cleanliness levels ‘In Operation’ cfu/m3 Area EU GMP FDA Guide USP Aseptic core <1 <3 Support for aseptic filling <10 <20 Controlled process area <100 Controlled support area <200 Not specified Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates Class Active air sample Settle plate Contact plate Swab Isolator or Closed RABS (ISO 5 or better) <0.1% ISO 5 <1% ISO 6 <3% ISO 7 <5% ISO 8 <10% Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates NOTE: Contamination recovery rates should be based upon actual monitoring data and should be re-tabulated monthly. When contamination recovery rates are observed that exceed the recommendations in the table or are greater than established process capability corrective actions should be taken. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates For example Grade B cleanroom Active air-sampling One year of data reviewed Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates Count (cfu) Frequency Cumulative percentage 1240 80.78% 1 191 93.22% 2 52 96.61% 3 20 97.92% 4 8 98.44% 5 7 98.89% 6 99.02% 99.22% 99.35% 9 99.41% 10 Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates Pharmaceutical Microbiology: http://www.pharmamicroresources.com

USP contamination rates Corrective actions may include but are not limited to: Sanitization program including types of disinfectants, application methods, and frequencies. Personnel practices by supervisory staff. Microbiological sampling methods and techniques. Training on gowning practices. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Frequencies of sampling Isolators Active Air Sampling-once/day Surface sampling-at end of each campaign Glove sampling-left to the users discretion Note differences to EU GMP for continuous monitoring. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Frequencies of sampling RABS Open RABS and closed RABS are different regarding contamination risk. Open RABS is more similar to a conventional clean room. Closed RABs are advanced aseptic processing systems. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Frequencies of sampling Cleanrooms Unchanged from the previous version of <1116> appearing in PF31(2). ISO class 5 = Each operating shift ISO class 7 = Each operating shift ISO class 8 = Twice per week Other areas = Once per week Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Sampling locations ISO 14644 grid approach for particles discussed, but dismissed “Microbiological sampling sites are best selected with consideration of human activity during manufacturing operations.” From careful observation and mapping of the clean room The most likely route of contamination is airborne Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Other changes / main features Strong emphasis upon staff training, including those who take microbiological samples Need for a qualified site microbiologist Staff health checks and control of entry to critical areas Importance of correct gowning Importance of risk assessment and risk mitigation Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Other changes / main features USP is strongly supportive of the use of “RMMs” in Environmental Monitoring and all forms of microbiological analysis. All references to validation have been removed from the chapter. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Regulatory differences Results: EU GMP Annex 1 discusses averaging of micro data. FDA mandates response to individual excursions. USP focuses on incident rates. Disinfection residues FDA and EU GMP suggest microbial resistance to sanitizers is possible. USP does not cover this. Media fills. All guidances use the same media fill criteria. USP provides no details for large fills. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Future developments Development of a chapter on Microbiological Control & Monitoring of Non-Aseptic Processing Environments <1111> has been discussed by USP MSA Problems: Operations vary much more widely than in aseptic processing. No widely accepted standards for the various facility designs. Significant differences in approach for the same product types. A clear path? Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Summary The challenge in aseptic processing is always personnel: as a source of microbial and particle contamination. Environmental monitoring continues to be especially contentious. The contamination rate concept is a significant departure from the alert and action level system that has been used in industry since the early 1980’s. Pharmaceutical Microbiology: http://www.pharmamicroresources.com

Pharmaceutical Microbiology: http://www.pharmamicroresources.com Thank you Pharmaceutical Microbiology: http://www.pharmamicroresources.com Pharmaceutical Microbiology: http://www.pharmamicroresources.com