Environmental Monitoring

Considerations
Nancy Roscioli, Ph.D.
Don Hill and Associates, Inc.

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

Environmental Monitoring
Components
Water monitoring:

Total organic carbon

Conductivity
Microbial Contaminants
Endotoxin

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

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

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)

Regulatory Basis for Environmental

Monitoring Program
CFR GMP regulations
FDA Guidance Documents
USP Informational Chapter

21 CFR 211.42
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

21 CFR 211.46
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.

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

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

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

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

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 > 0.5)

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

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

Viable and Nonviable Contaminant

Limits
Classifi- Nonviable (>0.5)
cation ft3
m3

Viable (CFU)
ft3

m3

Class
100

100

3,530

0.1

3.5

Class
10,000

10,000

353,000

0.5

18

Class
100,000 3,530,000 2.5
100,000

88

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

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

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.

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

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

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

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

Microbial Monitoring
Airborne viable contaminants
Surface contaminants

walls
equipment surfaces
countertops
floors

Personnel contaminants

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

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

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

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

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

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

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

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

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

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

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

Investigations and Corrective

Actions
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

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

Temperature and Humidity

Control of temperature and humidity
required for aseptic processing areas
21 CFR 211.42(c)(10)(ii)

Generally 65F and 35-50% humidity are

average
Too high - Increases personnel shedding
Too low - Increase static electricity

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

Water Requirements
Test
TOC
Conductivity
Micro.
Purity
EndoToxin

Potable Purified WFI

Water Water
none
500 ppb 500 ppb
none

See USP Table

500
CFU/ml

100
CFU/ml

none

none

10 CFU/
100 ml
0.25
EU/ml

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

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

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

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)

Water System Monitoring

Purified Water Systems
Weekly monitoring of system for:
microbial quality
TOC
conductivity

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

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

Water Use
Potable Water
Non-product contact uses
Feed for purified water system

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

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

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

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

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.