OOD AND RUG DMINISTRATION Revision #: 02 F D A Document Number: OFFICE OF REGULATORY AFFAIRS Revised: ORA.007 Office of Regulatory Science 25 Aug 2020 Title: Pharmaceutical Microbiology Manual Page 1 of 92 Sections in This Document 1. Purpose ................................................................................................................................. 1 2. Introduction ............................................................................................................................ 2 3. Current Editorial Board .......................................................................................................... 3 4. Original Authors ..................................................................................................................... 3 5. Chapter 1: Antimicrobial Effectiveness Testing ..................................................................... 4 6. Chapter 2: Microbial Examination of Non-Sterile Products ..................................................... 8 7. Chapter 3: Sterility Testing ................................................................................................. 14 8. Chapter 4: Investigating USP Sterility Testing Failure ......................................................... 28 9. Chapter 5: Bacterial Endotoxin Testing ................................................................................ 30 10. Chapter 6: Particulate Matter .............................................................................................. 40 11. Chapter 7: Antibiotic Potency Testing .................................................................................. 44 12. Chapter 8: Bioburden Estimation for Medical Devices......................................................... 59 13. Chapter 9: Environmental Monitoring .................................................................................. 61 14. Chapter 10: Inspectional Guidance ..................................................................................... 70 15. Glossary/Definitions ............................................................................................................. 90 16. Records ............................................................................................................................... 91 17. Supporting Documents ........................................................................................................ 91 18. Document History ................................................................................................................ 91 19. Change History .................................................................................................................... 92 20. Attachments ......................................................................................................................... 92 1. Purpose The purpose of this Pharmaceutical Microbiology Manual (PMM) is to collectively clarify, standardize, and communicate useful analytical procedures that are not specifically addressed in the microbiology methods chapters in the United States Pharmacopeia. In addition, some sections of this manual can serve as a technical reference when conducting microbiological inspections of drug, biotechnology and medical device manufacturers. The contents of this PMM were collaboration between ORS and CDER in order to maximize the efficiency of our analytical results to support CDER’s goal to assure the safety and reliability of commercially distributed medical products. For the most current and official copy, check QMiS. OOD AND RUG DMINISTRATION Revision #: 02 F D A Document Number: OFFICE OF REGULATORY AFFAIRS Revised: ORA.007 Office of Regulatory Science 25 Aug 2020 Title: Pharmaceutical Microbiology Manual Page 2 of 92 2. Introduction The Pharmaceutical Microbiology Manual (PMM) evolved from the Sterility Analytical Manual and is a supplement to the United States Pharmacopeia (USP) for pharmaceutical microbiology testing, including antimicrobial effectiveness testing, microbial examination of non-sterile products, sterility testing, bacterial endotoxin testing, particulate matter, device bioburden and environmental monitoring testing. The goal of this manual is to provide an ORS/CDER harmonized framework on the knowledge, methods and tools needed, and to apply the appropriate scientific standards required to assess the safety and efficacy of medical products within ORS testing laboratories. The PMM has expanded to include some rapid screening techniques along with a new section that covers inspectional guidance for microbiologists that conduct team inspections. This manual was developed by members of the Pharmaceutical Microbiology Editorial Board and includes individuals with specialized experience and training. The instructions in this document are guidelines for ORS analysts. When available, analysts should use procedures and worksheets that are standardized and harmonized across all ORS labs, along with the PMM, when performing analyses related to product testing of pharmaceuticals and medical devices. When changes or deviations are necessary, documentation should be completed per the laboratory’s Quality Management System. Generally, these changes should originate from situations such as new products, unusual products, or unique situations. This manual was written to reduce compendia method ambiguity and increase standardization between ORS laboratories. By providing clearer instructions to ORS labs, greater transparency can be provided to both industry and the public. However, it should be emphasized that this manual is a supplement and does not replace any information in USP or applicable FDA official guidance references. The PMM does not relieve any person or laboratory from the responsibility of ensuring that the methods being employed from the manual are fit for use, and that all testing is validated and/or verified by the user. The PMM will continually be revised as newer products, platforms and technologies emerge or any significant scientific gaps are identified with product testing. Reference to any commercial materials, equipment, or process in the PMM does not in any way constitute approval, endorsement, or recommendation by the U.S. Food and Drug Administration. For the most current and official copy, check QMiS. OOD AND RUG DMINISTRATION Revision #: 02 F D A Document Number: OFFICE OF REGULATORY AFFAIRS Revised: ORA.007 Office of Regulatory Science 25 Aug 2020 Title: Pharmaceutical Microbiology Manual Page 3 of 92 U.S. FDA, Office of Regulatory Affairs Office of Regulatory Science Office of Medical Products and Specialty Laboratory Operations 3. Current Editorial Board Editor-In-Chief: Angele Smith, ORS Headquarters Rick Friedman, Center for Drug Evaluation and Research Jennifer Gogley, Pacific Southwest Medical Products Laboratory Kristy Ford, Detroit Medical Products Laboratory Marilyn Khanna, ORS Headquarters Mivoyel Jeanpaul, ORS Headquarters Sammie La, Pacific Southwest Medical Products Laboratory Matthew Silverman, Winchester Engineering and Analytical Center Marla Stevens-Riley, Center for Drug Evaluation and Research Allison Rodriguez, Winchester Engineering and Analytical Center Radhakrishna Tirumalai, United States Pharmacopeia Theresa To, Winchester Engineering and Analytical Center 4. Original Authors Rhonda Alexander, Southeast Food and Feed Laboratory Jasna Braut-Taormina, Northeast Medical Products Laboratory Jennifer Brzezinski, Forensic Chemistry Center Marie B. Buen-Bigornia, Denver Food and Feed Laboratory Rick Friedman, Center for Drug Evaluation and Research Jennifer Gogley, Pacific Southwest Medical Products Laboratory Andrew Gonzales, Denver Food and Feed Laboratory Dennis Guilfoyle (ret), Northeast Medical Products Laboratory Lawrence James (ret), Northeast Food and Feed Laboratory Marilyn Khanna, ORS Headquarters David Lau, San Francisco Food and Feed Laboratory Eileen Liu, San Francisco Food and Feed Laboratory For the most current and official copy, check QMiS. OOD AND RUG DMINISTRATION Revision #: 02 F D A Document Number: OFFICE OF REGULATORY AFFAIRS Revised: ORA.007 Office of Regulatory Science 25 Aug 2020 Title: Pharmaceutical Microbiology Manual Page 4 of 92 Mercedes Loftis, Denver Food and Feed Laboratory Philip McLaughlin (ret), Winchester Engineering and Analytical Center Zachary Miller, Denver Food and Feed Laboratory Gary Pecic, Denver Food and Feed Laboratory Diane Raccasi (ret), Office of Enforcement and Import Operations George Salem (ret), ORS Headquarters Angele Smith, ORS Headquarters Tammara Stephens, Southeast Food and Feed Laboratory Selen Stromgren, ORS Headquarters Vilasini Suktankar, Southeast Food and Feed Laboratory Radhakrishna Tirumalai, United States Pharmacopeia Evelyn Wong, Pacific Southwest Medical Products Laboratory Jonathon Yenovkian, San Francisco Food and Feed Laboratory 5. Chapter 1: Antimicrobial Effectiveness Testing Antimicrobial Effectiveness testing is described in USP <51>. Previously this chapter was known as “Preservative Effectiveness Testing”. Detailed procedure for the performance of the test can be found in USP <51>. A. Media For the cultivation of the test organisms, select agar medium that is favorable to the rigorous growth of the respective stock culture. The recommended media are Soybean Casein Digest Agar/Broth and Sabouraud’s Dextrose Agar/Broth. Add a suitable inactivator (neutralizer) for the specific antimicrobial properties in the product to the broth and/or agar media used for the test procedure whenever