Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
In-House Microbial
Isolates in Compendial
Testing
Regulatory Requirements
1
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
2
Agenda
 Current Industry Debate Over the Use...
3
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Current Industry Debate Over the Use
of In-Ho...
4
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
"Good practice includes the periodic
challeng...
5
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
PMF List (2010)
 “As soon as an environmenta...
6
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
The arguments against the inclusion of in-hou...
7
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Does an “environmental” isolate lose its “wil...
8
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
“It’s too difficult to be practical” does not...
9
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
10
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Are 483 observations and Warning Letters
lim...
11
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Disinfectant Efficacy Testing
Methodology
12
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Point of clarification …
 “Disinfectant”: A...
13
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 1: Determine the test method
◦ Tube m...
14
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 2: Determine the Challenge
Organisms
...
15
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 3: Determine the Surface Types to
be ...
16
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 4: Determine Expiration of
Disinfecta...
17
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 5: Method Validation
◦ Critical!
◦ Ty...
18
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Step 6: Efficacy Testing
◦ USP <1072>, “Di...
19
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
20
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Disinfectant Efficacy Testing
Compliance Def...
21
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Disinfectant effectiveness studies
agains...
22
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “The firm was unable to provide scientific...
23
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Disinfectant Efficacy Testing
Industry Guides
24
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Routinely used disinfectants should be ef...
25
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
United States Pharmacopeia, <1072> “Disinfec...
26
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “It is a sound practice to perform challen...
27
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Disinfectant Efficacy Testing
Industry Liter...
28
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Validation of sanitising agents for effec...
29
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “… users are expected to challenge disinfe...
30
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “…scientifically-based references and
conv...
31
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Disinfectant testing involves challenging...
32
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Media Growth Promotion Testing
Methodology
33
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 USP <61>, “Microbiological Examination
of ...
34
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Challenge the medium with a low-level
inoc...
35
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Media Growth Promotion Testing
Compliance De...
36
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Not limited to aseptic manufacturers!
Manu...
37
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “… the growth promotion tests for media
us...
38
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Growth promotion testing performed on med...
39
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Growth promotion studies were deficient be...
40
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 The FDA expressed its satisfaction that
th...
41
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Media Growth Promotion Testing
Industry Guid...
42
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “The QC laboratory should determine if USP...
43
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “… microorganisms used in growth-
promotio...
44
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
From PDA Technical Report No. 22, “Process S...
45
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
From the Pharmaceutical Inspection
Conventio...
46
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Media Growth Promotion Testing
Industry Lite...
47
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Growth promotion should be demonstrated u...
48
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “In addition to the compendial organisms r...
49
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “In addition to type cultures, environment...
50
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Test Method Suitability/Qualification
Compli...
51
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 There has been no documented evidence that...
52
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Test Method Suitability/Qualification
Indust...
53
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “There is a strong argument that environme...
54
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Rapid Microbiological Methods
Compliance Def...
55
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Rapid microbiological methods (RMMs)
have ...
56
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “You should develop a panel of microorgani...
57
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Rapid Microbiological Methods
Industry Liter...
58
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “To demonstrate the [RMM’s] ability to det...
59
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Validation studies evaluated sterility te...
60
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Selection of In-House Isolates for Use
in Co...
61
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Depends upon the test itself, and upon
the...
62
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 21 CFR 211.113(a) : “Appropriate written
p...
63
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
“Case Studies of Microbial Contamination in ...
64
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “HACCP [Hazard Analysis Critical Control P...
65
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Since the microbial quality of pharmaceut...
66
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Design of the manufacturing facility
and e...
67
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Environmental monitoring program
◦ Charact...
68
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 PDA Technical Report No. 13,
“Fundamentals...
69
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “A well-established microbial EM program m...
70
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 “Although drugmakers have begun to perform...
71
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Selection of In-House Isolates for Use
in Co...
72
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Two bases for selection of isolates
◦ Demo...
73
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Disinfectant efficacy testing
◦ Microflora...
74
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Example …
 EM trend reports describe histor...
75
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Example …
 Introduction of an “objectionabl...
76
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Example …
 EM excursion
 Data trend analys...
77
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Subjecting the organisms to disinfectant
e...
78
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Inclusion of in-house isolates in media
gr...
79
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Media used for raw material and
component ...
80
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Media used for in-process product testing
...
81
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Test method suitability/qualification
◦ Em...
82
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
 Sources of information useful for selectin...
83
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Review
 Current Industry Debate Over the Us...
84
In-House Microbial Isolates in Compendial
Testing: Regulatory Requirements
Robert Westney
Director of Quality and Opera...
Upcoming SlideShare
Loading in …5
×

In-House Microbial Isolates in Compendial Testing: Regulatory Requirements

6,913 views

Published on

This presentation provides an overview of the current regulatory expectations for the use of in-house microbial isolates in compendial testing. It reviews regulatory, compendial and industry references on the topic. Importantly, it also provides a strategy for selection of these isolates.

Published in: Education

In-House Microbial Isolates in Compendial Testing: Regulatory Requirements

  1. 1. In-House Microbial Isolates in Compendial Testing Regulatory Requirements 1
  2. 2. In-House Microbial Isolates in Compendial Testing: Regulatory Requirements 2 Agenda  Current Industry Debate Over the Use of In- House Isolates in Compendial Testing  Disinfectant Efficacy Testing  Media Growth Promotion Testing  Test Method Suitability/Qualification  Rapid Microbiological Methods  Selection of In-House Isolates for Use in Compendial Testing
  3. 3. 3 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Current Industry Debate Over the Use of In-House Isolates in Compendial Testing
  4. 4. 4 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements "Good practice includes the periodic challenge of prepared media with low levels of organisms. This includes USP indicator organisms as well as normal flora." U.S. Food and Drug Administration: “Guide to Inspections of Microbiological Pharmaceutical Quality Control Laboratories” (1993)
  5. 5. 5 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements PMF List (2010)  “As soon as an environmental isolate is put onto growth media, it is no longer an ‘environmental’ isolate’.”  “EM [environmental monitoring] organisms will stop to behave as such after a few transfers.”  “Environmental isolates are often mismanaged by the microbiology lab.”  “Many companies simply don't have the qualified equipment and personnel to preserve and maintain stock cultures of their environmental bugs.”  “This is one of the problems with using 483 observations as ‘cGMP’. They are not. They are the observations of one inspector who may or may not be knowledgeable in microbiology.”
  6. 6. 6 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements The arguments against the inclusion of in-house isolates fall into three categories: 1. An “environmental” isolate, once cultured in the Microbiology laboratory, loses its “wild-type” traits. 2. Preparing an in-house microbial isolate to create the organism concentration necessary for compendial testing is too problematic to be practical. 3. The regulatory expectation that in-house microbial isolates be included in compendial testing is represented by only a few “rogue” FDA inspectors, and is not an Agency-wide expectation.
  7. 7. 7 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Does an “environmental” isolate lose its “wild-type” traits after being cultured?  There has been no published empirical evidence that an in-house microbial isolate either does or does not lose its “wild-type” traits upon culturing.  United States Pharmacopeia (USP <61>): “Seed-lot culture maintenance techniques (seed-lot systems) are used so that the viable microorganisms used for inoculation are not more than five passages removed from the original master seed-lot”.  Although in most cases an in-house isolate likely becomes physiologically more “robust” upon culturing, it is reasonable to assume that a limited number of passages from the source culture allows the microorganism to retain its “wild-type” genetic traits.
  8. 8. 8 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements “It’s too difficult to be practical” does not address the scientific merit of including in-house isolates in compendial testing.  The basic process includes preparation of a suspension of unknown concentration, performing serial dilutions and plate counts to determine the concentration, and then standardizing the suspension to the desired concentration.  During the incubation period for serial dilution plates, the suspension is typically held under refrigerated conditions for several days.  During this time, the concentration of the organism in suspension may change, resulting in either a decrease or increase.  Standardization and final use of this suspension may therefore result in a “miss” in terms of its assumed concentration, discovered only after testing is complete.
  9. 9. 9 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements
  10. 10. 10 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Are 483 observations and Warning Letters limited to just a few inspectors?  “Compliance versus science”  The increasing trend in citing firms for not meeting this requirement demonstrates that the Agency’s SMEs recognize the scientific merit of including in-house isolates in compendial testing.  Defending against this expectation is a challenge!
  11. 11. 11 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Disinfectant Efficacy Testing Methodology
  12. 12. 12 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Point of clarification …  “Disinfectant”: A chemical agent used on inanimate objects (e.g., floors, walls, sinks) to destroy virtually all recognized pathogenic microorganisms, but not necessarily all microbial forms (e.g., bacterial endospores). [Centers for Disease Control and Prevention]  “Sanitizer”: A chemical agent that simultaneously cleans and disinfects.
  13. 13. 13 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 1: Determine the test method ◦ Tube method: Inoculate dilutions of the disinfectant and determine the microbial reduction. Typically referred to as “use- dilution” testing. ◦ Coupon method: Inoculate the surfaces of coupons representing facility materials of construction. Expose them to disinfectant. Remove the inoculum. Determine log reduction. "Six Steps to Qualifying Disinfectants“, Deborah Ensign, Microtest Laboratories, Inc., 2011
  14. 14. 14 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 2: Determine the Challenge Organisms ◦ American Type Culture Collection (ATCC) organisms (gram-negative, gram-positive, spore-former, fungi) ◦ Environmental isolates
  15. 15. 15 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 3: Determine the Surface Types to be Tested ◦ Examples: stainless steel, glass, plastic, tile, plexiglas ◦ 2” x 2” coupons ◦ Sterilize or disinfect before using.
  16. 16. 16 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 4: Determine Expiration of Disinfectants ◦ Use the same concentration and contact exposure time used in the facility. ◦ Use the worst-case expiration date for each disinfectant.
  17. 17. 17 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 5: Method Validation ◦ Critical! ◦ Typically called the “recovery” part of the test ◦ Must demonstrate adequate recovery of the challenge organisms in the presence of the disinfectants. ◦ Inoculate the test system (i.e., the medium exposed to disinfectant) with a low-level challenge of test organism. Include a control that has not been exposed. ◦ Recovery: 50-200% of control
  18. 18. 18 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Step 6: Efficacy Testing ◦ USP <1072>, “Disinfectants and Antiseptics” ◦ Typically called the “challenge” part of the test. ◦ Inoculate tube/coupon. ◦ Expose to the desired concentration of the disinfectant for the desired contact time. ◦ Demonstrate 2-log reduction for spore-formers. ◦ Demonstrate 3-log reduction for vegetative bacteria.
  19. 19. 19 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements
  20. 20. 20 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Disinfectant Efficacy Testing Compliance Deficiencies
  21. 21. 21 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Disinfectant effectiveness studies against representative microorganisms and/or specific in-house isolates were not conducted for cleaning agents used in your facility to disinfect production areas, including aseptic areas.” [FDA Warning Letter, March 28, 2008]  “No rationale was provided for the ATCC [American Type Culture Collection] organisms used nor was [sic] actual EM isolates used for the study.” [FDA Memorandum, Biologics License Application, 2009]
  22. 22. 22 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “The firm was unable to provide scientific rationale for the use of the selected organisms used in the Disinfectant Efficacy study. These organisms were not representative of organisms isolated from the facility nor were they representative of the USP guidelines.” [FDA 483 Observation, May 25, 2011]  “Your response states that a supplemental disinfectant efficacy study, using mold spores of in house isolates on various surfaces, will be performed and completed by [redacted]. We suggest that this proposed study be conducted as soon as possible.” [FDA Warning Letter, July 12, 2012]
  23. 23. 23 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Disinfectant Efficacy Testing Industry Guides
  24. 24. 24 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Routinely used disinfectants should be effective against the normal microbial vegetative flora recovered from the facility... If indicated, microorganisms associated with adverse trends can be investigated as to their sensitivity to the disinfectants employed in the cleanroom in which the organisms were isolated.” [FDA Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing - Current Good Manufacturing Practice, September 2004]  “If using an Association of Official Analytical Chemists (AOAC) method, microorganisms specified in the reference which are most likely to be found in the manufacturing environment should be used and tests performed on microbial isolates frequently found in the environment can provide additional information on the effectiveness of disinfectants.” [“Regulatory Aspects Concerning the Quality Controls of Microbiological and Nonviable Particulate Contamination in Pharmaceutical Manufacturing”, Oji, (FDA), American Pharmaceutical Review, January/February 2004]
  25. 25. 25 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements United States Pharmacopeia, <1072> “Disinfectant and Antiseptics”  “… the most frequently isolated microorganisms from an environmental monitoring program may be periodically subjected to use-dilution testing with the agents used in the disinfection program to confirm their susceptibility, as there are real differences among different species in resistance to the lethal effects of different sanitizers.”  “To demonstrate the efficacy of a disinfectant within a pharmaceutical manufacturing environment, it may be deemed necessary to conduct the following tests: (1) use-dilution tests (screening disinfectants for their efficacy at various concentrations and contact times against a wide range of standard test organisms and environmental isolates); (2) surface challenge tests (using standard test microorganisms and microorganisms that are typical environmental isolates, applying disinfectants to surfaces at the selected use concentration with a specified contact time, and determining the log reduction of the challenge microorganisms); and (3) a statistical comparison of the frequency of isolation and numbers of microorganisms isolated prior to and after the implementation of a new disinfectant.”
  26. 26. 26 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “It is a sound practice to perform challenge testing of the selected sanitizers/disinfectants with isolates routinely recovered by the environmental monitoring program. This establishes the practical effectiveness of the disinfectants.” [Parenteral Drug Association Technical Report No. 13, “Fundamentals of an Environmental Monitoring Program”, September/October 2001]  “Regulatory authorities now expect evidence of the efficacy of disinfection agents against environmental isolates.” [ASTM E2614 – 08, Standard Guide for Evaluation of Cleanroom Disinfectants]
  27. 27. 27 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Disinfectant Efficacy Testing Industry Literature
  28. 28. 28 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Validation of sanitising agents for effectiveness against isolated organisms found in cleanroom environments is increasingly becoming an area of concern to both manufacturers and regulatory agencies.” [“Surface Disinfectant Validations”, Nelson Laboratories, Inc., 2004]  “It is generally recommended to use fungal spore suspensions of both reference cultures and environmental isolates... Regulatory authorities also expect to see the specific environmental isolates most frequently found in the facility included in the disinfectant effectiveness testing.” [“Control Strategies for Fungal Contamination in Cleanrooms”, Lopolito et al., Controlled Environments Magazine, September 2007]
  29. 29. 29 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “… users are expected to challenge disinfectants not only with standard test organisms but also with facility environmental isolates because commercially available microorganisms behave quite differently from their ‘wild’ counterparts. Selection of the test organisms is crucial and an important issue to the regulatory agencies, especially when it comes to environmental isolates.” [“Microbial Limit and Bioburden Tests: Validation Approaches and Global Requirements”, Clontz, October 14, 2008]  “...it is crucial to make an educated investment in antimicrobial products that are effective against your specific mold isolates.” [“Evaluating the Activity of Disinfectants Against Fungi”, Polarine et al., CleanRooms, Volume 23, No. 2, February 2009]
  30. 30. 30 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “…scientifically-based references and conventional wisdom used to select these products does not alleviate drug manufacturers of the requirement to validate disinfectants, sporicides, and even isopropyl alcohol for use in facilities under actual use conditions against environmental isolates.” [“Prevention of Microbial Contamination“, Kopis Sartain and Polarine, Pharmaceutical Technology, June 2, 2011  “Disinfectant Efficacy Studies should be considered as a fundamental requirement during the development of qualifications of disinfectants on various surfaces with both ATCC and in-house isolates.” [Friedman, Blog, August 15, 2011]
  31. 31. 31 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Disinfectant testing involves challenging the disinfectant solution (as a suspension test) and challenging different surface materials with disinfectant and a range of different microorganisms including isolates from the facility.” [ “Cleanroom Cleaning and Disinfection: Eight Steps for Success”, Sandle, Controlled Environments Magazine, March 2012]  “This study clearly demonstrates that the most frequently isolated microorganisms from an environmental monitoring program may be periodically subjected to microbroth dilution testing with cleanroom disinfectant agents used in the disinfection program to confirm their sensitivity profile.” [“In vitro Antifungal Efficacy of Biguanides and Quaternary Ammonium Compounds against Cleanroom Fungal Isolates”, Vijayakumar et al., PDA Journal of Pharmaceutical Science and Technology, May/June 2012]
  32. 32. 32 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Media Growth Promotion Testing Methodology
  33. 33. 33 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  USP <61>, “Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests”  USP <62>, “Microbiological Examination of Nonsterile Products: Tests for Specified Organisms”  USP <71>, “Sterility Tests”
  34. 34. 34 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Challenge the medium with a low-level inoculum of test organism: <100 cfu  Control = Previously qualified medium  ATCC organisms  In-house isolates  Agar: 50-200% recovery  Broth: Turbidity
  35. 35. 35 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Media Growth Promotion Testing Compliance Deficiencies
  36. 36. 36 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Not limited to aseptic manufacturers! Manufacturers of non-sterile products have been subject to regulatory enforcement.
  37. 37. 37 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “… the growth promotion tests for media used during environmental monitoring did not include the use of the normal microbial flora commonly recovered and isolated from the various production and support areas.” [FDA Warning Letter, February 24, 1997]  “Growth promotion qualification of the media used for environmental monitoring does not include a challenge with mold isolates.” [FDA Warning Letter, July 10, 1998]
  38. 38. 38 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Growth promotion testing performed on media fill vials does not include evidence the media is capable of detecting environmental isolates found in class 100 filling areas. For example, mold organisms are not used to challenge media, even though mold isolates have been identified in filling room 1.” [FDA Warning Letter, January 11, 2001]  “… your firm does not perform challenge testing to the sterility media with environmental isolates from the environmental monitoring program.” [FDA Warning Letter, October 29, 2010]
  39. 39. 39 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Growth promotion studies were deficient because there was “no inclusion of environmental isolates in the growth promotions that are conducted, including growth promotion studies for aseptic media simulations.” [FDA 483 Observation, May 2008]  “… the firm does not test TSA … during growth promotion tests for microorganisms to include for example, molds, yeasts and other potential known environmental contaminants found in the manufacturing facility and/or raw materials.” [FDA 483 Observation, April 30, 2010]
  40. 40. 40 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  The FDA expressed its satisfaction that the firm complied with its expectation: “Growth promotion studies were conducted successfully using indicator microorganisms per USP as well as local isolates.” [FDA Memorandum, BLA Recommendation, January 16, 2009]
  41. 41. 41 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Media Growth Promotion Testing Industry Guides
  42. 42. 42 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “The QC laboratory should determine if USP indicator organisms sufficiently represent production-related isolates. Environmental monitoring and sterility test isolates can be substituted (as appropriate) or added to the growth promotion challenge.” [FDA Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing - Current Good Manufacturing Practice, September 2004]  “… for the Growth Promotion test, representative microflora isolated from the controlled environment or ATCC strain preparations of these isolates may also be used to test media.” [USP <1116>, “Microbiological Evaluation of Clean Rooms and Other Controlled Environments”]
  43. 43. 43 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “… microorganisms used in growth- promotion testing may be based on the manufacturer's recommendation for a particular medium, or may include representative environmental isolates.” [USP <1117>, “Microbiological Best Laboratory Practices”]  USP clarifies parenthetically that “these latter are not to be construed as compendial requirements.”
  44. 44. 44 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements From PDA Technical Report No. 22, “Process Simulation for Aseptically Filled Products” (2011):  “The growth promotion properties of the incubation media should be evaluated using pharmacopeial methods. The inclusions of tests for environmental organisms or those isolated from sterility test positives are recommended.”  “Growth promotion testing ... is performed using pharmacopeial methods. Consideration should be given to testing with other microorganisms found in the aseptic processing area environment, such as those isolated during environmental and personnel monitoring and sterility test contaminants.”
  45. 45. 45 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements From the Pharmaceutical Inspection Convention/Pharmaceutical Inspection Cooperation Scheme (PIC/S):  “The medium selected should be capable of supporting a wide range of microorganisms, which might reasonably be encountered and be based also on the in house flora (e.g. isolates from monitoring etc.).” [Recommendation on the Validation of Aseptic Processes. PI-007-6, 1 January 2011]  “Environmental or fastidious organisms may be used if alternative non-selective enrichment media have been selected for the sterility test.” [Recommendation on Sterility Testing. PI-012-3, 25 September 2007]
  46. 46. 46 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Media Growth Promotion Testing Industry Literature
  47. 47. 47 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Growth promotion should be demonstrated using organisms listed in USP General Chapter <71> as well as environmental, personnel, and sterility test failure isolates [at the] <100-cfu [colony-forming unit] challenge.” [“Microbial Testing in Support of Aseptic Processing”, Cundell, Pharmaceutical Technology, June 2004]  “In general, a microbial growth medium such as soybean casein digest medium should be used. This media selected should be demonstrated to promote growth of United States Pharmacopoeia (USP) <71> indicated organisms as well as representative isolates identified from environmental monitoring, personal monitoring, and positive sterility test results.” [Environmental Monitoring for Cleanrooms and Controlled Environments, Dixon (Ed.), November 2, 2006.]
  48. 48. 48 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “In addition to the compendial organisms required in the [media growth promotion] tests, addition of specific microorganisms of interest could be useful if they have been recovered from past tests (e.g., a Sterility Test contaminant or a frequent environmental monitoring isolate).” [“Quality Control of Microbiological Culture Media”, Sutton, PMF Newsletter, January 2006]  “The media must be challenged with the organisms listed in the current version of the compendia as well as one or more in-house isolates… The in-house cultures used must be qualified as well, with regards to identity and population size. The challenge should be with <100 CFU and the population size must be verified and recorded.” [“Introduction to Sterility Testing: Control of the Environment, Test Limitations, and Investigating Manufacturing Systems”, Mentel, American Pharmaceutical Review, Jan/Feb 2006]
  49. 49. 49 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “In addition to type cultures, environmental isolates are commonly used in media testing regimes. These organisms are designed to demonstrate that a particular lot of culture media will grow microorganisms which are representative of the types that are found in the manufacturing environment... The use of such isolates is increasingly becoming a regulatory expectation.” [“Assessment of Culture Media in Pharmaceutical Microbiology”, Sandle, Pharmaceutical Microbiology Blog, April 3, 2012]
  50. 50. 50 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Test Method Suitability/Qualification Compliance Deficiencies Industry Guides
  51. 51. 51 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  There has been no documented evidence that the FDA has cited a manufacturer for lack of inclusion of in-house isolates in microbiological test method suitability/qualification.  “The test organisms selected should reflect organisms that could be found in the product, process, or manufacturing environment.” [FDA Final Rule on “Amendments to Sterility Test Requirements for Biological Products”, June 4, 2012]  Recent Rule… more regulatory scrutiny in the near future!
  52. 52. 52 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Test Method Suitability/Qualification Industry Literature
  53. 53. 53 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “There is a strong argument that environmental isolates are the best challenge to media and for validation studies like sterility test validation. They are the most sensitive micro-organisms, having been exposed recently to disinfectants, particular soils etc.” [“The Use of Environmental Isolates”, Sandle, Pharmaceutical Microbiology Blog, January 10, 2010]  “The author highly suggests performing test method suitability studies using wild-type isolates from production surfaces instead of laboratory-adapted strains of bacteria. Wild-type strains are a better representation of the organisms encountered on production areas than those strains that lack wild characteristics.” [“Bioburden Method Suitability for Cleaning and Sanitation Monitoring: How Far Do We Have to Go?”, Salaman-Byron, Pharmaceutical Technology, August 2, 2010]
  54. 54. 54 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Rapid Microbiological Methods Compliance Deficiencies Industry Guides
  55. 55. 55 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Rapid microbiological methods (RMMs) have become a growing area of advancement over the past several years.  There has been no documented evidence that the FDA has cited a manufacturer for lack of inclusion of in-house isolates in rapid microbiological methods.
  56. 56. 56 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “You should develop a panel of microorganisms relevant to the product and process to challenge the performance of your RMM. We recommend that you include in your panel microorganisms which represent the following categories: … ◦ isolates detected in starting materials, ◦ isolates detected by in-process testing or during preliminary product testing, ◦ isolates detected by environmental monitoring of your manufacturing facility, ◦ isolates from your production areas which represent low nutrient and high stress environments …” FDA Guidance for Industry: Validation of Growth-Based Rapid Microbiological Methods for Sterility Testing of Cellular and Gene Therapy Products (Draft), February 2008
  57. 57. 57 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Rapid Microbiological Methods Industry Literature
  58. 58. 58 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “To demonstrate the [RMM’s] ability to detect a range of microorganisms, at least six replicate samples of drug product were spiked with 10 to 100 cfu of microorganisms from site-specific isolates and … compendial microorganisms … ” [“The Introduction of Qualitative Rapid Microbiological Methods for Drug-Product Testing”, Newby et al., Pharmaceutical Technology, Process Analytical Technology, 2004]
  59. 59. 59 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Validation studies evaluated sterility test samples from primary culture, expansion culture, and final product culture with a microbial challenge comprised of 10 microbial species. These species included a mix of commercial reference strains and stressed cells from frozen archives of environmental isolates and sterility test failures. During the validation process, FDA recommended challenge microorganisms … ” [“Automated Rapid Microbiological Methods for the Biopharmaceutical Industry: Selection, Validation, and Implementation for an Autologous Cell Therapy Product”, Duguid & du Moulin, American Pharmaceutical Review, June 2009]  “Additional considerations [for RMM validation] may be provided with regard to ... environmental isolates.” [“Revision of PDA Technical Report Number 33”, Miller and Moldenhauer, American Pharmaceutical Review, February 1, 2010] PDA Technical Report 33, Evaluation, Validation and Implementation of New Microbiological Testing Methods, May 2000
  60. 60. 60 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Selection of In-House Isolates for Use in Compendial Testing Risk
  61. 61. 61 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Depends upon the test itself, and upon the nature of the manufacturing process and final product.  Risk ◦ Assessment ◦ Management ◦ Mitigation
  62. 62. 62 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  21 CFR 211.113(a) : “Appropriate written procedures, designed to prevent objectionable microorganisms in drug products not required to be sterile, shall be established and followed.”  21 CFR 211.113(b): “Appropriate written procedures, designed to prevent microbiological contamination of drug products purporting to be sterile, shall be established and followed. Such procedures shall include validation of all aseptic and sterilization processes.”
  63. 63. 63 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements “Case Studies of Microbial Contamination in Biologic Product Manufacturing”, Suvarna et al., American Pharmaceutical Review, 2011.
  64. 64. 64 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “HACCP [Hazard Analysis Critical Control Point] might be used to identify and manage risks associated with physical, chemical, and biological hazards (including microbiological contamination).” ICH Q9, “Quality Risk Management”, 2005]  “Where warranted, a risk-based assessment of the relevant factors [regarding recovery of unspecified microorganisms] is conducted by personnel with specialized training in microbiology and in the interpretation of microbiological data. For raw materials, the assessment takes account of the processing to which the product is subjected, the current technology of testing, and the availability of materials of the desired quality.” [USP <1111> “Microbiological Examination of Nonsterile Products: Acceptance Criteria for Pharmaceutical Preparations and Substances for Pharmaceutical Use”]
  65. 65. 65 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Since the microbial quality of pharmaceutical products is a function of microorganisms introduced through excipients, the significance of microorganisms recovered from excipients should be evaluated in terms of the number and type of organisms, whether or not they will survive the manufacturing process, their ability to grow in the product formulation, the use of the product and the potential hazard to the user.” [“Microbiological Considerations When Selecting Excipients During Product Development”, Montgomery and Manu-Tawiah, American Pharmaceutical Review, 2004]
  66. 66. 66 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Design of the manufacturing facility and environmental monitoring (EM) program  “Where microbiological specifications have been established for the intermediate or API, facilities should also be designed to limit exposure to objectionable microbiological contaminants, as appropriate.” [ICH Q7A, “Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients”, 2000]
  67. 67. 67 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Environmental monitoring program ◦ Characterization of normal facility microflora ◦ Identifying shifts in microflora, seasonal trends  PDA Technical Report No. 13, “Fundamentals of an Environmental Monitoring Program”, September/October 2001 ◦ “Characterizing microorganisms recovered from environmental and personnel monitoring is an important part of surveillances programs” ◦ “… routine characterization should continue to determine whether isolates are part of the normal microbial flora or represent something different.”
  68. 68. 68 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  PDA Technical Report No. 13, “Fundamentals of an Environmental Monitoring Program”, September/October 2001 ◦ “A change in the microbial flora or the introduction of a previously undetected species might signify a change in a system that should be investigated” ◦ “Characterizations can be useful clues as to the possible source of isolates.”
  69. 69. 69 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “A well-established microbial EM program must identify, at least at the genus level, microorganisms isolated from samples that reach the alert and action limits established. Evaluation of the isolates is necessary and would be evaluated for their ability to grow in the non-sterile product present at the time of sampling. It is necessary to make a list of microorganisms isolated and identified at least at the genus level during the first year of implementation. This list is crucial to identify the profile of microorganism species found in production areas, its possible origin, trends, and seasonal peaks.” [“Bioburden Method Suitability for Cleaning and Sanitation Monitoring: How Far Do We Have to Go?”, Salaman-Byron, Pharmaceutical Technology, 2010]
  70. 70. 70 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  “Although drugmakers have begun to perform environmental monitoring in their nonsterile manufacturing facilities, many companies are uncertain about how to interpret the data and develop alert and action levels … The industry should record the types and numbers of microorganisms that it identifies in its environmental-monitoring programs and evaluate the information on a quarterly basis … This strategy could provide guidance, based on scientific rationale, for assessing and controlling the risk of contamination.” [“Manufacturers Recalibrate Microbial Control for Nonsterile Drugs”, Greb, Pharmaceutical Technology, Equipment and Processing Report, February 16, 2011]
  71. 71. 71 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Selection of In-House Isolates for Use in Compendial Testing Strategies
  72. 72. 72 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Two bases for selection of isolates ◦ Demonstrating adequate reduction of a high- level presence of an organism  Disinfectant efficacy testing (“challenge”) ◦ Demonstrating adequate recovery of a low- level presence of an organism  Disinfectant efficacy testing (“recovery”)  Media growth promotion testing  Test method suitability/qualification  Rapid Microbiological Methods
  73. 73. 73 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Disinfectant efficacy testing ◦ Microflora of the manufacturing environment ◦ Typically evaluated in EM trend reports ◦ Undesirable trends (e.g., seasonal) ◦ Recovery of organisms considered “objectionable” or “of concern”
  74. 74. 74 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Example …  EM trend reports describe historical and consistent seasonal “blooms” of filamentous fungi (mold) during the autumn season.  Corrective action = frequent sanitization with a sporicidal agent  Characterize (ID) the species of mold(s).  Include the mold(s) in disinfectant efficacy testing.  Importantly, the early recovery of the species (detected through EM) signals the need to proactively implement frequent sanitization with a sporicidal agent.  Don’t react to the “bloom” … Risk mitigation!
  75. 75. 75 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Example …  Introduction of an “objectionable” organism into the facility ◦ Recurring incident? ◦ Ingress into more stringently classified areas as a result of material, personnel or equipment flow?  Conclusion ◦ Adverse trend? ◦ Shift in the facility’s normal microflora?  Include the organism in disinfectant efficacy testing.
  76. 76. 76 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Example …  EM excursion  Data trend analysis  Conclusion = adverse trend  Include the organism(s) in disinfectant efficacy testing.
  77. 77. 77 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Subjecting the organisms to disinfectant efficacy testing … ◦ Lack of efficacy demonstrates the need to change disinfectant ◦ Demonstrating efficacy implicates inadequate cleaning procedures/practices  Include the organisms in EM media growth promotion testing.
  78. 78. 78 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Inclusion of in-house isolates in media growth promotion testing ◦ Media used for EM and clean utility monitoring ◦ Media used for raw material, component, in- process product and finished product testing
  79. 79. 79 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Media used for raw material and component testing ◦ Previously recovered “objectionable” or fastidious organisms from non-sterile materials ◦ Previously recovered contaminants from sterile materials
  80. 80. 80 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Media used for in-process product testing ◦ Review of historical data ◦ Critical control points  Media used for finished product testing ◦ For non-sterile products, the approach is similar to that of selecting isolates from non- sterile raw materials and components. ◦ For sterile products, include isolates recovered from previous sterility test or media fill failures.
  81. 81. 81 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Test method suitability/qualification ◦ Employ the same strategy as that used for selection of isolates for growth promotion of media used for raw material, component, in- process product and finished product testing.  “Objectionable” and fastidious organisms  Sterility failures  Media fill failures
  82. 82. 82 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements  Sources of information useful for selecting in-house isolates ◦ Annual product reviews ◦ EM and clean utility trend reports ◦ Process validation ◦ Cleaning validation ◦ Media fills  Organisms representing risk
  83. 83. 83 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Review  Current Industry Debate Over the Use of In- House Isolates in Compendial Testing  Disinfectant Efficacy Testing  Media Growth Promotion Testing  Test Method Suitability/Qualification  Rapid Microbiological Methods  Selection of In-House Isolates for Use in Compendial Testing
  84. 84. 84 In-House Microbial Isolates in Compendial Testing: Regulatory Requirements Robert Westney Director of Quality and Operations Cryologics, Inc. (610) 847-8781 rwestney@cryologics.com www.Cryologics.com Links to references: http://www.cryologics.com/resources/bibliography

×