The document discusses proposed changes to Annex 1 regarding viable monitoring of cleanrooms. It notes that Annex 1 will now require continuous monitoring throughout critical processes, rather than interval sampling. This reflects a shift from air sampling to real-time air monitoring. Continuous active air monitors are presented as able to meet the new Annex 1 requirements, capturing transient events better than settle plates. The document discusses various active air monitoring devices and their particle collection capabilities compared to traditional samplers. Real-time microbial monitoring using laser induced fluorescence is also proposed as an alternative to growth-based methods.

1. 1
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John Cobb
Microbiology Manager
A NEW APPROACH TO ACTIVE AIR SAMPLING, MEETING THE PROPOSED
CHANGES TO ANNEX 1 REGARDING VIABLE MONITORING

2. 2
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• Brief Introduction to the Speaker – John Cobb
• EM and Regulations
• Microbial “Monitoring” of Air
• Annex 1 Draft – Our interpretation!

3. 3
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Design and validation of active air samplers
Development of Irradiated Media to GMP Standards
My background

4. 4
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Real-Time and Continuous Microbial Monitoring Systems

5. 5
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WHY DO WE DO ENVIRONMENTAL MONITORING?

6. 6
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Regulations………of course!
• Applicable regulations for Sterile Manufacturingin the EU market and
Monitoring of Operating Theatres in Hospitals:
• UNI-EN-ISO 14644/1-2-3 for cleanroom classification and testing.
• UNI-EN-ISO 14698/1-2 for biocontamination control in cleanroom.
• Pharmacopoeias: USP, JP, EP
• HTM 03-01 referring to Microbiological Commissioning and Monitoring of
Operating Theatre Suites
• Annex 1 of the EU Guide to Good Manufacturing Practice (Orange Guide)
(2003)
• Annex 1 Consultation Document Draft (2017)

7. 7
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The E.C. Guideline to GMP recommends limits for Environmental Monitoring of clean
areas during operation (2002,revised in Nov 2008)

8. 8
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Let’s be critical of where we are at!!
Is following the minimum requirements of
current Regulations enough to help you
understand what is happening in your
critical areas?

9. 9
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Why do we take interval samples when
performing an “Active Air Sampling” regime?
Should we microbially monitor over an entire
production run to gain a more reliable
understanding of our critical areas?

10. 10
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Current Criticisms about
Active Air Sampling as Part of an EM
Programme for Grade A Areas

11. 11
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Typical Human Upper Respiratory Flora
BACTERIUM Nose Pharynx Mouth
Staphylococcus epidermidis ++ ++ ++
Staphylococcus aureus* + + +
Streptococcus mitis + ++
Streptococcussalivarius ++ ++
Streptococcus mutans* + ++
Streptococcus pneumoniae* +/- + +
Streptococcus pyogenes + +
Neisseria sp. + ++ +
Neisseria meningitidis + ++ +
Proteus sp. + + +
Haemophilus influenzae* + + +
Lactobacillus sp. + ++
Corynebacteria ++ + +
Actinomycetes + +
Spirochetes + ++
Mycoplasmas + +
Green: Will grow
on TSA/TSB
Red: Will NOT
grow* on TSA/TSB
Orange: May
Grow on TSA/TSB
*Under typical
incubation conditions.
“Regulatory Expectations for Aseptically
Produced Parenterals”
Ian Symonds, Director AsepticQuality
Assurance , GlaxoSmithKline
December 2009PDA Meeting, Milan

12. 12
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Current Sampling
•Filter Area = 6m2
•Air Rate = 0.45m/S
•Volume Of Air In 1min =162m3
•Volume Of Air, 1hour = 9,720m3
•Volume Of Air, 8hr Shift = 77,760m3
• Active Air Sample = 4m3 0.005 % Air
• Continuous Particle Monitoring = 16m3 0.02 % Air
• Surface Monitoring;
–10 Contact Plates = 0.0250m2 = 0.417 % Surface*
–10 Swabs = 0.0250m2 = 0.417 % Surface
• Based upon the footprint of the filling machine area within the RABS. Note: the actual total surface area would be
much higher

13. 13
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Bottom Line on Current Growth Based Methods
• Only a small fraction of air and surfaces influencing
product quality can be tested
• Monitoring techniques have low recovery efficiency
• A limited range of organisms can be isolated using
common media and incubation conditions
• A positive isolate is a significant event
• A negative result may be misleading

14. 14
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Proposed Changes to Annex 1
Where is the industry going with regards to
Environmental Monitoring during Manufacturing in
Grade A Areas?

15. 15
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Annex 1 of the GMP Guide to the Manufacture of Sterile Medicinal Products (the “Orange Guide”) is changing
the way environmental monitoring of viable organisms performed from routine sampling to continuous
monitoring.
9.27 Continuous monitoringin grade A and B areas should be undertaken for the full duration of critical processing,
including equipment(aseptic setup) assemblyand filling operations (i.e., an understanding of function and interactions of
each clean area). The monitoring should beperformed in such a way that all interventions, transientevents and any
system deterioration would becaptured and any riskcaused by interventions of themonitoringoperations is avoided.
9.28 Rapid microbial monitoring methods maybe adopted after validation as long as they are demonstrated to be at least
equivalent to theestablished methodology.
9.33 If microorganisms aredetected in a gradeA or B zone, they should beidentified to species level and theimpact of
such microorganismson product quality(for each batch implicated) and state of controlshould be evaluated.
Consideration mayalso begiven to theidentification of gradeC and Dcontaminantsand therequirements should be
defined in thecontaminationcontrol strategy.
In addition, ISO 14698 specifies some more air sampler specific criteria for active air sampling:
• Must entrap particles down to 1µm
• Sample at least 1m³
• Operate within 1 foot of critical area
• Settle plates can be used for up to 4 hours
As monitoring was not required throughout the process,
sampling was usually performed before the start and after
the end of a process. This was seen as ‘down time’ in the
process which lead to an array of rapid cubic metre air
samplers becoming available. These are samplers not
monitors!
The Need for Monitoring not
Sampling

16. 16
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Annex 1 Proposed Revision
(under section Viable Monitoring)
Settle Plates or Active Air Sampler??

17. 17
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Settle Plates

18. 18
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Settle Plates “count” the number of vegetative
microbial particles that are heavy enough to
displace from the air onto a surface, in a 4 hour
period.
• What is that telling us?
• Is that enough information?
• Smaller particles?

19. 19
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MONITORING BY CONTINUOUS ACTIVE AIR SAMPLING

20. 20
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ImpactAir®
140
ISO-140® ISO-90®
Annex 1
Compliant Solutions

21. 21
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ImpactAir®and ISO-140®
Environmental Monitoring Solutions
• ImpactAir & ISO-140– Air Monitors
• Fully validated for ISO 14698-1 for Physical & Biological Efficiency
• Class leading d50 = 0.42µ
• Unique RELIABLE dynamic height adjustment*
• Consistent d50 throughout run*
• Proven monitoring technology
• Up to 4hrs or 6.8m³ per plate
• Does not shed particles
• HEPA filtered exhaust
• Robust stainless Steel construction
• Ultra low vibration
• Low power (<60 wats)
• Optional remote control unit (RCU)
• Time correlated results
• Drop in replacement for Air Trace
*IP protected
ISO-140 & ISO-CON
Controller

22. 22
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ISO-90® –
Critical Location Monitoring
• Designed to Monitor Critical Locations
• 4 hours on 9cm agar plate
• Fully validated for ISO 14698-1 for Physical & Biological Efficiency
• d50 <0.75µ
• 125mm diameter
• Proven monitoring technology
• Does not shed particles
• HEPA filtered exhaust – environment
• 9cm Agar plate
• Robust stainless Steel construction
• Ultra low vibration
• Low power
• Time correlated results
In current development is a ground-breaking ultra small monitor designed to monitor
critical locations. It draws air in at a slower rate to minimise air flow disturbance yet
meeting all of the requirements of Annex 1 whilst using a 9cm agar plate.

23. 23
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d50 Values of Available
Samplers & Monitors
Monitor
d50 = 0.42
Sampler
d50 =1.1
Sampler
d50 =1.51
Sampler
d50
=1.1/1.61
Sampler
d50 = 1.61
Sampler
d50 =0.78
Sampler
d50 =1.11/1.61
Monitor
d50 = 0.42
Sampler
d50 = 21.5
Monitor
d50 = <0.75
Monitor
d50 = <0.75
EnvironmentMonitors Critical Location Monitors
These are Samplers NOT Monitors

24. 24
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ImpactAir….

25. 25
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AN ALTERNATIVE METHOD:
REAL-TIME MICROBIAL MONITORING USING LASER
INDUCED FLOURESENCE (LIF)

26. 26
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What Microbial contamination might we consider to be in the
Environment?
“Sterile and Non-Sterile Product Risks
From Viable But Non-Culturable Bacteria”
Edward C. Tidswell PhD
Senior Director
Baxter Healthcare Corporation
October 2009, PDA 5th Annual Global
Conference on Pharmaceutical Microbiology,
Washington, DC

27. 27
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Current Environmental Airborne Monitoring Technology
Up to 5 Days
Particle
Excursion
Response

28. 28
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No methods are perfect….

29. 29
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Microbial Monitoring in Real-Time

30. 30
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31. 31
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What is the BioTrak and how does it work?
3 instruments in 1

32. 32
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BIOTRAK®: Total Particle Counting

33. 33
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BIOTRAK®: Viable Particle Counting
Viability Detector
Laser Induced Fluorescence
(LIF)

34. 34
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BIOTRAK®: Microbial Collection

35. 35
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BioTrak:
Real-Time Aerosol Optical Spectroscopy
Scattered Light
Fluorescence A
Fluorescence
B
What happens when you try to determine viability using 3
parameters?
3 Parameters: Can
discriminate !

36. 36
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How does LIF compare to classical
microbiology…

37. 37
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Comparability
Culture
Media
Culturable
Organism
Viable Cell Metabolites Growth Based
VBNC
Viable But Non
Culturable
Collection
Efficiency
Presence of
Viability
Fluorophores
Fluorescing
Nonviable

38. 38
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How do we use Real-Time
Monitors…

39. 39
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40. 40
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Increase Confidence and Process Understanding
Applications Benefits
• Root cause investigations
• Replace air samplers, settle
plates and particle counters
• Isolators/aseptic core
• FMS alerts/alarms
• Real-Time
• Intervention Free
• Continuous Monitoring
• Reduce risk
• Enhance manufacturing

41. 41
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REAL-TIME MICROBIAL MONITORING AND THE REGS

42. 42
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9.28 Rapid microbial monitoring methods may
be adopted after validation as long as they are
demonstrated to be at least equivalent to the
established methodology.
9.29 Sampling methods should not pose a risk
of contamination to the manufacturing
operations.
Revised Annex 1
Rapid Methods

43. 43
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Real-Time Viable Particle
Counters and the Regs
• Annex 1 Draft
• PEMM (Process Environmental Monitoring Methods)
working group
• BioPhorum Group (Air)
➢ Real-time contamination risk-management in drug productmoves a
step closer
• Validation
• USP<1223>, EP 5.1.6 and TR 33 mention the implementation
and validation of rapid methods
• USP<1116> mentions the importance of trending and
process control

44. 44
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What do PMT Supply?

46. 46
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IQ OQ PQ
On site trial
Supporting
documents
Evaluation
report
Service/Calibration
Validation
assistance

47. 47
www.pmtgb.com
Germany
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Benelux
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France
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Thanks for your time.. Any questions
John Cobb
www.pmtgb.com
JohnCobb@pmtgb.com
Visit us at Stand 513
Ericsson Hall 1