1. MONITORING CONTROLLED ENVIRONMENTS Pacific BioLabs Inc. (510) 964-9000 info@PacificBioLabs.com

2. 2 CONTROLLED AREA Manufacturing area where non-sterile product and in process material contact equipment surfaces or are exposed to the environment Viable and nonviable contaminants are controlled to specific levels Class 100,000 and Class 10,000

3. 3 CRITICAL AREA Aseptic processing area where sterile products/components are exposed to the environment and no further processing will occur Class 100

4. 4 ROOM CLASSIFICATION (CLASS NAMES) ISOUS FS 209EUSP SI 31M 1.5 410M 2.5 5100 M 3.5 61,000M 4.5 710,000M 5.6 8100,000M 6.5

5. 5 ROOM CLASSIFICATION LIMITS IN PARTICLES > 0.5µm ISOUS FS 209EISO (m 3 )FS 209E (ft 3 ) 3135.21 41035210 51003,520100 61,00035,2001,000 710,000352,00010,000 8100,0003,520,000100,000

6. 6 BUILDING Sufficient space to allow  proper cleaning, maintenance, and manufacturing functions  orderly operations  contamination control Sealed windows, flush surfaces Changing rooms/washing facilities

7. 7 BUILDING (cont.) Clean utilities such as gasses, water HVAC system Filtration of air – HEPA’s Airflow from critical to less critical areas Air lock to maintain positive pressure

8. 8 ENVIRONMENT/HVAC SYSTEM VALIDATION HVAC air velocity, airflow patterns HEPA filter integrity and efficiency Air pressure differentials  0.04 to 0.06 inches of water gauge Cleaning and sanitization/disinfection studies Airborne non-viable particle counts Airborne viable particle counts

9. 9 EXAMPLE ( www.fda.gov/cdrh/qsr/06bldng.html) Specifications for a medical device assembly facility Class10,000 Particles > 0.5µmGuess ?? Air Pressure0.05 inches of water Temperature72 + 2.5°F Air Velocity90 feet/minute

10. 10 REGULATORY BASIS FOR ENVIRONMENTAL MONITORING CFR GMP regulations FDA Guidance Documents USP Informational Chapter

11. 11 ENVIRONMENTAL CONTROL 21 CFR 820.70 (c) “Where environmental conditions could reasonable be expected to have an adverse effect on product quality, the manufacturer shall establish and maintain procedures to adequately control these environmental conditions”

12. 12 ENVIRONMENTAL MONITORING COMPONENTS Non-Viable Particles  Air Microbial Contamination  Air  Surface Pressure Differential Water quality Temperature and Humidity

13. 13 PRODUCT BIOBURDEN Does not have to be part of an environmental monitoring program Test performed on a non-sterile product to determine its microbial load Reflects the quality control of manufacturing process and raw materials Needed to verify adequacy of sterilization process

14. 14 ENVIRONMENTAL CONTROL 21 CFR 820.70 (c) An uncontrolled environment may result in inconsistent bioburden levels Bioburden spikes may exceed the sterilization process capability to achieve the desired SAL

15. 15 MICROBIAL IDENTIFICATION USP  An environmental monitoring program should include identification of the flora obtained from sampling. ANSI/AAMI/ISO TIR 15843:2000  Characterization of bioburden is required to reduce the frequency of dose audits

16. 16 ENVIRONMENTAL MONITORING PROGRAM Documented in SOP Details procedures used for monitoring Includes sampling sites Specifies sampling frequency Describe investigation when Alert or Action levels are exceeded Describes methods for trend analysis Training

17. 17 AIRBORNE PARTICULATE COUNT AKA total particulate count Detection of particles > 0.5 µm (outside of US particles > 5.0 µm are counted) Monitoring is recommended during operations Optical particle counting equipment is commonly used

18. 18 MICROBIAL MONITORING Assess the effectiveness of sanitization practices and of personnel Provides sufficient information to ascertain that the environment is controlled Is conducted during normal operations

19. 19 MICROBIAL MONITORING Room air Compressor air Surfaces  Equipment  Sanitization containers  Floors  Walls  Personnel garments

20. 20 Airborne Viable Particulate Count - Methods Passive monitoring  Settling plates  Not generally recommended in US Active monitoring  Solid culture medium impaction  Testing of known volumes of air that allow quantification by unit of volume air

21. 21 AIRBORNE VIABLE PARTICULATE COUNT - EQUIPMENT Passive air monitoring  Petri dish with agar Active air monitoring  Slit-to-Agar (STA)  Sieve Impactors  Centrifugal Impactors  Filtration  Liquid Impingement  Gelatin Filter Sampler

22. 22 SURFACE MICROBIAL MONITORING METHODS Contact Plates Flexible Films Swabs Surface Rinse Methods

23. 23 PERSONNEL MONITORING Garments  Chest  Sleeves  Other areas are sampled for qualification Gloves  Finger impressions

24. 24 EXAMPLE OF SAMPLING SITES SystemSite Environmental air (filling)Near open containers Room airProximal to work areas WaterPoint of use Surface (facility)Floor, door handles, walls Surface (equipment)Filling line, control panels Compressed airFarthest from compressor Laminar air flowNear high activity areas OperatorFinger impressions

25. 25 SAMPLING FREQUENCY Sampling AreaFrequency Class 100 or lessEach shift Class 10,000Each shift Some support areasTwice/week Product/container contact areasTwice/week Other support areas > Class 100,000 Once/week

26. 26 TRAINING PROGRAM Personal hygiene/habits Illness Clothing/gowning practices Introduction to microbiology GMPs Introduction to aseptic techniques Participation in media fills to demonstrate aseptic skill level Must be documented

27. 27 ALERT AND ACTION LEVELS Alert Level  A level than when exceeded indicates a process may have drifted from its normal operating condition. Warning that does not warrant a corrective action Action Level  A level than when exceeded indicates a process has drifted from its normal operating condition. Documented investigation and corrective action required

28. 28 AIR - ACTION LEVELS ClassCFU/m 3 CFU/ft 3 100< 3< 0.1 10,000< 20< 0.5 100,000< 100< 2.5

29. 29 EQUIPMENT/FACILITIES SURFACE – ACTION LEVELS ClassCFU per Contact Plate 1003 (including floor) 10,0005 10 (floor)

30. 30 PERSONNEL GEAR SURFACE – ACTION LEVELS Class Gloves (cfu/plate) Clothing (cfu/plate) 10035 10,0001020

31. 31 ACTION LEVEL INVESTIGATIONS Review of:  Maintenance records  Sanitization documentation  Operational parameters Identification of microbial contaminants Training of personnel

32. 32 CORRECTIVE ACTIONS Training of personnel Additional sampling Increased frequency of sampling Additional sanitization Additional product testing Evaluation of the need to revise SOPs Product impact/disposition documented

33. 33 WATER REQUIREMENTS TestWFIPurifiedPotable TOC500 ppb None ConductivitySee USP None Microbial10 CFU/100mL100 CFU/mL500 CFU/mL Endotoxin0.25 EU/mLNone

34. 34 WATER SYSTEM MONITORING FREQUENCY TestWFI SystemPurified Water EndotoxinDaily *None MicrobialDaily *Weekly TOCWeekly ConductivityWeekly

35. 35 ENVIRONMENTAL MONITORING SURVEILLANCE SUPPORT Alert and Action Levels Data Management  Collection, trend analysis and interpretation Isolates Characterization Investigation/Corrective Actions Documentation

36. 36 REFERENCES “Fundamentals of Environmental Monitoring”, Supplement TR 13, PDA J. Pharm. Sci. & Tech. 55(6), 2001. United Stated Pharmacopeia 30, Microbiological Evaluation of Clean Rooms and Other Controlled Environments. The United States Pharmacopeia Convention Inc., Rockville, MD. pp 589-596 (2007). United Stated Pharmacopeia 30, Pharmaceutical Compunding-Sterile Preparations. The United States Pharmacopeia Convention Inc., Rockville, MD. pp 334-351 (2007). United States Food and Drug Administration “Medical Device Quality Systems Manual” (January 1997).

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