Environmental-Monitoring

1. Environmental Monitoring
Considerations

2. Environmental Monitoring
Components
• Airborne nonviable particulate monitoring
• Airborne viable contaminant monitoring
• Viable contaminant monitoring of surfaces
• Viable contaminant monitoring of personnel
• Temperature and humidity monitoring
• Pressure differential monitoring

3. Environmental Monitoring
Components
• Water monitoring:
– Total organic carbon
– Conductivity
– Microbial Contaminants
– Endotoxin

4. General Environmental
Monitoring Considerations
• Monitoring frequencies and strategies
– Establishment of a meaningful and manageable
program
• Sampling and testing procedures
• Establishment of effective alert and action
limits
• Trending of results

5. General Environmental
Monitoring Considerations
• Investigation and evaluation of trends as
well as excursions from alert and action
limits
• Corrective actions to be implemented in
response to environmental monitoring
excursions
• Personnel training - sampling, testing,
investigating excursions, aseptic technique

6. Scope of Environmental
Monitoring Program
• Should include monitoring of all
environments where products and their
components are manufactured
– All areas where there is a risk of product
contamination
• Should include monitoring of all water used
for product manufacturing as well as feed
water to the final water purification system
(WFI System)

7. Regulatory Basis for Environmental
Monitoring Program
• CFR GMP regulations
• FDA Guidance Documents
• USP Informational Chapter

8. • Aseptic processing areas:
– Easy to clean and maintain
– Temperature and humidity controlled
– HEPA filtered air
– Environmental monitoring system
– Cleaning and disinfecting procedures
– Scheduled equipment maintenance and
calibration

9. • Ventilation, air filtration, air heating and
cooling:
– Adequate control over microorganisms, dust,
humidity and temperature.
– Air filtration systems including prefilters and
particulate matter air filters for air supplies to
production areas.

10. Guideline on Sterile Drug Products
Produced by Aseptic Processing
• Defines critical and controlled
manufacturing areas
• Recommends airborne nonviable and viable
contaminant limits
• Provides some guidance on monitoring
frequencies for critical areas

11. Guideline on Sterile Drug Products
Produced by Aseptic Processing
• Recommendations for air pressure
differentials
• Includes guidance on aseptic media fills
• Note: This guidance document was written
in 1987 and is in need of revision

12. Microbial Evaluation and Classification
of Clean Rooms and Clean Zones
• USP General Information Chapter <1116>
• Establishment of clean room classifications
– Federal Standard 209E
• Importance of EM program
• Personnel training in aseptic processing
• Establishment of sampling plans and sites
– suggested sampling frequencies

13. Microbial Evaluation and Classification
of Clean Rooms and Clean Zones
• Establishment of alert and action limits
• Suggests limits for airborne, surface and
personnel contaminant levels.
• Methods and equipment for sampling
• Identification of isolates
• Aseptic media fills
• Emerging technologies - barrier; isolator

14. Federal Standard 209E
• “Airborne Particulate Cleanliness Classes in
Clean Rooms and Clean Zones
• Approved by the GSA for use by all Federal
Agencies
• Frequently referenced for controlled
environment particulate requirements:
Classes 100, 10,000 and 100,000 (based on
particles > 0.5µ)

15. Guidance for Industry for Sterile Validation
Process Validation in Applications for Human
and Veterinary Drug Products
• Scope limited to final drug product manufacturing
and data required for application submission
(NDA, BLA)
• Requests information on:
– Buildings and facilities
– Manufacturing operations for drug product
• Filter validation
• Validation of hold times

16. Guidance for Industry for Sterile Validation
Process Validation in Applications for Human
and Veterinary Drug Products
• Requests information on:
– Sterilization and depyrogenation
– Media fills and actions taken when they fail
– Microbiological monitoring of the environment
• Airborne microorganisms, personnel, surfaces,
water system, product component bioburden
– Yeasts, molds, anaerobes
– Exceeded EM limits

17. Viable and Nonviable Contaminant
Limits
Nonviable (>0.5µ) Viable (CFU)
Classifi-
cation ft3
m3
ft3
m3
Class
100
100 3,530 0.1 3.5
Class
10,000
10,000 353,000 0.5 18
Class
100,000
100,000 3,530,000 2.5 88

18. Controlled Area
• Preparation or manufacturing area where
nonsterile product, in-process materials and
product-contact equipment surfaces,
containers and closures are exposed to the
environment
• Control nonviable and viable contaminants
to reduce product /process bioburden
• Class 100,000 or Class 10,000

19. Controlled Area
• Capping areas are now considered
controlled manufacturing areas
– Should be supplied with HEPA filtered air
– Should meet class 100,000 conditions during
static conditions

20. Critical Area
• Aseptic processing area where sterile
products, components or in-process
products are exposed to the environment
and no further processing will occur.
• Air quality must be Class 100 during
processing
• Local Class 100 areas are often utilized
during open processing steps during drug
substance manufacture.

21. Critical Area
• The area just preceding the sterile core
should be one classification higher than the
core.

22. Nonviable Particulate Monitoring
• Airborne cleanliness classifications should
be met during operations
• Nonviable monitoring should occur
routinely during operations
• Monitoring during static conditions is done
as part of HVAC qualification and may be
done periodically after that to insure area
meets acceptable conditions before use or
following cleaning

23. Nonviable Particulate Monitoring
• Locations for monitoring should be
established during performance
qualification; probes placed close to work
surface
• Monitoring frequencies vary:
– For aseptic processing areas, during each use
– For other, controlled areas, varies from each use
to weekly or less depending on use of area

24. Nonviable Particulate Monitoring
• HVAC Validation and Maintenance
Considerations:
– Air velocity, airflow patterns and turbulence
should be validated; smoke studies to determine
flow patterns during static and dynamic
conditions
– HEPA filter integrity testing
– HEPA filter efficiency testing
– Air pressure differentials

25. Microbial Monitoring
• Airborne viable contaminants
• Surface contaminants
– walls
– equipment surfaces
– countertops
– floors
• Personnel contaminants

26. Microbial Monitoring
• Monitoring methods should be capable of
detecting molds and yeasts
• Should also be able to detect anaerobes
– Most often, this is an issue associated with
products filled anaerobically (with nitrogen
overlay)
• All lots of media for EM sampling should
be growth promotion tested

27. Microbial Monitoring
• Routine microbial monitoring should take
place during operations (for airborne
contaminants) and immediately following
operations (for surfaces and personnel).
• Airborne monitoring frequencies:
– Each use for aseptic processing areas
– Varies from daily to weekly to less frequently
for controlled areas depending on use

28. Microbial Monitoring
• Personnel and surface monitoring
frequencies vary:
– Aseptic processing - after every fill
– Other controlled areas - varies from daily to
weekly or less for surfaces
– Personnel monitoring often restricted to aseptic
area personnel and personnel working in Class
100 hoods performing tasks such as inoculation

29. Microbial Monitoring
• Monitoring of surfaces and airborne
contaminants during rest periods (following
cleaning)
– Important for confirming adequacy of cleaning
procedures
– Indicates whether HVAC system is operating
properly
– NOTE: Disinfectant effectiveness studies also
required for cleaning agents used in the facility

30. Microbial Monitoring
• Monitoring frequencies and procedures are
influenced by a number of factors:
– Stage of manufacturing
– “Open” or “closed” manufacturing step
– Single or multiple product manufacturing

31. Microbial Monitoring
• Establishment of monitoring locations
should be based on performance
qualification studies during dynamic
conditions
– gridding study to determine worst case
locations/most meaningful locations
• Should also establish common flora - will
aid in investigations

32. Setting Alert and Action Limits
• Action limits (for the most part) have been
established in a variety of guidance
documents
• Alert limits
– Lower than action limits
– Reflect actual historical results under normal
processing conditions

33. Exceeding Limits
• Alert limits are designed to provide some
warning that environmental quality is
approaching action limit and allow you time
to correct.
• Exceeding alert limit triggers a warning
response - i.e., alert affected area personnel
• Exceeding multiple alerts - triggers action
level response

34. Exceeding Limits
• Action limit excursions require
investigations
– Speciation of organism(s)
– Review batch records from date of excursion
– Review other recent EM data (trends)
– Review cleaning records
– Interview personnel
– Product impact - must quarantine until
determined

35. Exceeding Limits
• Excursions from action limits require
corrective actions that may include:
– More rigorous or additional monitoring
– More rigorous cleaning
– Retraining of personnel
– Procedural changes - change to or addition of
disinfection procedures, for example
– HVAC maintenance

36. Investigations and Corrective
Actions
• The investigation procedures to be followed
should be pre-established and included in
SOPs
• Depending on the outcome of the
investigation, corrective actions should be
pre-established to the extent possible

37. • Imperative that EM results be linked to
product release so that affected products are
not released until investigation completed
• Material Review Board or equivalent should
be consulted prior to releasing product that
was potentially affected by adverse
environmental conditions
Investigations and Corrective
Actions

38. Trending
• Should trend monitoring results
(environmental and water)
– Periodic (quarterly or monthly) review by QA
and others
– Re-evaluation of action and alert limits on an
annual basis
– This trending information is generally included
in the Annual Product Review

39. Temperature and Humidity
• Control of temperature and humidity
required for aseptic processing areas
– 21 CFR 211.42(c)(10)(ii)
• Generally 65°F and 35-50% humidity are
average
– Too high - Increases personnel shedding
– Too low - Increase static electricity

40. Temperature and Humidity
• Temperature should be controlled
throughout all manufacturing areas
• Temperature and humidity should be
monitored and controlled in warehouse
areas where temperature/humidity sensitive
raw materials are stored
– If not able to control humidity, need procedure
to follow if humidity exceeds limit

41. Water Requirements
Test Potable
Water
Purified
Water
WFI
TOC none 500 ppb 500 ppb
Conduc-
tivity
none See USP Table
Micro.
Purity
500
CFU/ml
100
CFU/ml
10 CFU/
100 ml
Endo-
Toxin
none none 0.25
EU/ml

42. Water For Injection
• Defined by USP
• Water purified by distillation or reverse
osmosis
• Prepared from water complying with the
U.S. EPA National Primary Drinking Water
Regulations
• Contains no added substance

43. Purified Water
• Defined in USP
• Obtained by a suitable process, usually one
of the following:
– deionization
– reverse osmosis
– combination

44. Potable Water
• Meets National Drinking Water Regulations
• 40 CFR Part 141
• Periodic monitoring in-house as well as
periodic certificates from municipality (if
applicable)

45. Water System Monitoring
• WFI Systems
– Microbial quality and endotoxin
• Daily system monitoring
• Each use point at least weekly
– TOC and Conductivity
• Weekly system monitoring
• can be taken from worst case point (end of loop,
return to tank)

46. Water System Monitoring
• Purified Water Systems
– Weekly monitoring of system for:
• microbial quality
• TOC
• conductivity

47. Water Use
• WFI
– Solvent for preparation of parenteral solutions
– Formulation of mammalian cell culture media
– Formulation of purification buffers
– Final product formulation
– Vial and stopper washing
– Final rinse for product equipment

48. Water Use
• Purified Water
– Preparation of terminally sterilized
microbiological media
– Initial rinsing/cleaning
– Laboratory use
– Feed for WFI system

49. Water Use
• Potable Water
– Non-product contact uses
– Feed for purified water system

50. Microbial Monitoring Devices
• Slit-to-Agar (STA) - Powered by vacuum,
air taken in through a slit below which is a
slowly revolving plate.
• Sieve impactor - Vacuum draws in air
through perforated cover which is impacted
onto petri dish containing nutrient agar

51. Microbial Monitoring Devices
• Centrifugal Sampler - consists of a
propeller that pulls a known volume of air
into the unit and then propels the air
outward to impact on a nutrient agar strip
• Sterilizable Microbiological Atrium
(SMA)- similar to sieve impactor; cover
contains uniformly spaced orifices; vacuum
draws in air which is impacted on agar plate

52. Microbial Monitoring Devices
• Surface Air System Sampler - An
integrated unit containing an entry section
with an agar contact plate; behind is a motor
and turbine that pulls air in through the
perforated cover and exhausts it beyond the
motor.
• Settle plates - qualitative; may be useful in
worst case locations

53. Microbial Monitoring Devices
• Surface contaminant monitoring devices:
– Contact Plates - plates filled with nutrient
agar; for regular surfaces
– Swabs - useful for hard to reach or irregular
surfaces; swab placed in suitable diluent and
inoculated onto microbiological plate

54. Monitoring Considerations
• Remote sampling probes - validate use of
tubing
• Must sample adequate quantity of air to be
statistically meaningful.
– 80-100 ft3/min
• Must validate growth promotion after
exposure of settle plates (or other plates) for
prolonged time periods.