Environmental

Control and Measurement

Mark J. Stannard
Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

2 2
What is contamination?

 Presence of any unwanted substance in the

product
 Two types of contaminants
 Viable
 Non-viable

3 3
Why prevent contamination?

 Ensure product quality

 Protect our customers
 Comply with cGMP
regulations and laws
– 21 CFR 211 and 600
– EU GMP and Annex 1

4 4
Sources of Contamination?

 Water
 Air
 Surfaces
 People

5 5
Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

6 6
How is contamination controlled?

 Health and Hygiene

 Validated Sterilization of Equipment and Components
 Protective Apparel
 Aseptic Techniques
 Clean Room Conduct
 Cleaning and Disinfection Techniques
 Facility and Equipment Maintenance
 Open vs. Closed System Technology
 Air Flow, Filtration, and Pressurization

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Air Flow and Pressurization

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 HEPA Filtration and Pressurization

HEPA filtration and laminar flow serve as contamination control

devices. HEPA (High Efficiency Particulate Air) filters are used to remove
particulates and microorganisms from the air supply to the
manufacturing/filling rooms, laminar flow hoods, biosafety cabinets, etc.

Cross-section of a HEPA Filter

HEPA Fact: HEPA filters are made of boron silicate microfibers formed into a
flat sheet by a process similar to papermaking. The flat sheets are pleated to
increase the overall surface area. A HEPA filter is able to trap 99.99% of
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particles of a diameter greater than or equal to 0.3 microns. 9
Open System Technology

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 Closed System Technology
 Bulk Closed Systems
– Provide engineered solution to
achieve an advanced aseptic
processing environment.

 Benefits include
– Separation of people and their
contaminants from the aseptic
process by enclosing the filling
area, thus requiring that
personnel interventions, work,
and handling of materials be
conducted remotely.
– Use of contiguous piping to
improve the maintenance culture
purity/asepsis in the process.
– Use of clean in place and
sterilize in place systems for
vessel/piping decontamination

11 11
 Closed System Technology
 Barrier Isolator Technology
– Provides engineered solution to
achieve an advanced aseptic
processing environment.

 Benefits include
– Separation of people and their
contaminants from the aseptic
process by enclosing the filling
area, thus requiring that personnel
interventions, work, and handling of
materials be conducted remotely.
– Use of validated pass-through
device designs to improve the
maintenance of asepsis in the
isolator.
– Use of gas/vapor decontamination
agents with validated
decontamination cycles
The Mini Aseptic Filling System (MAFS) from Bosch-TL

12 Systems
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Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

13 13
Environmental Monitoring Program for
Classified Areas

 HEPA filter certification

 Qualification of Classified Areas
 Air Flow Visualization Studies
 Process Simulation Testing
 Routine Monitoring of Classified Areas and
Utilities
 Personnel Monitoring
 Test Processing and Result Analysis
 Investigations for Excursions
 Closed System Technology
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HEPA Filter Certification

 Equipment utilizing HEPA filter technology

must be certified initially and periodically.
 Certification tests include:
– a velocity profile
– induction leak test
– particle counts
– airflow pattern test
– a HEPA filter leak test
HEPA Filter

15 15
 Area Classification for Style-ogen®
Process Steps

•Tank Fermentation
•Non Sterile Purification
•Buffer/Media Hold Unclassified Space

Open Processing
Closed Equipment

Grade A

•Solution Prep
•Equipment Assembly Classified Space
•Sterile Purification

• Open Sterile Sampling

• Open Aseptic Transfers • Open Sterile Transfers

16
Lighter shading signifies more stringent area 16
classification
 Classified and Aseptic Areas

 Classified and Aseptic Areas

– Maximum allowable number of viable and non
viable particles per volume of air sampled
– Defined by regulatory agencies

Class 100,000/Grade C Gowning Processing

Class 10,000/Grade B Gowning
Room Room
Room Hall
Class 100/Grade A

Air
Air
Lock LFH
Lock

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 FDA/ISO Particulate and Microbial Air
Classificationsa and Levels

Clean Area ISO > 0.5 µm Microbiological Microbiological

Classification Designation b particles/m3 Active Air Action Settling Plates Action
(0.5 um Levels c (cfu/m3 ) Levels c,d (diam.
particles/ft3) 90mm; cfu/4 hours)

100 5 3,520 1e 1e
1000 6 35,200 7 3
10,000 7 352,000 10 5
100,000 8 3,520,000 100 50
a- All classifications based on data measured in the vicinity of exposed materials/articles during periods of activity.
b- ISO 14644-1 designations provide uniform particle concentration values for cleanrooms in multiple industries. An
ISO 5 particle concentration is equal to Class 100 and approximately equals EU Grade A.
c- Values represent recommended levels of environmental quality. You may find it appropriate to establish alternate
microbiological action levels due to the nature of the operation or method of analysis.
d- The additional use of settling plates is optional.
e- Samples from Class 100 (ISO 5) environments should normally yield no microbiological contaminants.

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Qualification of Classified Areas

 Initial Qualification
– Ensure HVAC systems are performing to specifications and
meeting regulatory levels for environmental control
– Select test sites based upon area of room (i.e., ISO formula)
and risk of contamination to product
– Air and Surface testing conducted for multiple days to ensure
satisfactory performance
 Periodic Re-Qualification
– Demonstrate continued satisfactory HVAC performance and
continued compliance with regulatory specified levels.
– At periodic intervals (e.g., semi-annually for Class 100/10,000;
annually for Class 100,000)
– Following area modifications and/or breaches of room integrity

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 Air Flow Visualization Studies

 Verify
– Airflow pattern characteristics and unidirectional nature of
airflow in the Grade A zone under static and dynamic conditions
– LFH’s ability to limit dispersion of viable/non-viable particles
 Assess
– Appropriate work flow and personnel aseptic technique specific
to the equipment design and airflow patterns
– Non-unidirectional occurrences, such as back-flow, ‘pulsing’ of
air, or dead spots.
– Optimal environmental monitoring test site locations for
qualification and routine testing.

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Air Flow Visualization Studies

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 Process Simulation (Media
Challenges)

 Simulates aseptic process using growth promoting

media in the place of actual product.
 Simulates process as closely as possible while
incorporating selected worst case conditions
 Assures that aseptic process conditions and
associated controls are sufficiently rigorous to ensure
the manufacture of pure culture or sterile product.
 Serves as a continuing assessment of the processes,
equipment, procedures, and personnel associated
with aseptic manufacturing.
 Conducted on predefined frequency as specified by
regulations
 Conducted following modifications/shutdowns
 Incubate media, examine for presence of
contamination (e.g., flocculation or turbidity)

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 Process Simulations (Cont.)

 Aseptic Filling
– Twice per year per line per
shift
– Min 5,000 vials filled
– Target 0 contaminated vials
– Perform all representative
interventions
– Aseptic PQ of Personnel

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Routine Monitoring: Air and
Compressed Gas Testing

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Routine Monitoring: Air and
Compressed Gas Testing (Cont.)

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Routine Monitoring: Surface Testing

 RODAC (Replicate Organism Detection

and Count) plates contain growth
promoting medium
Direct contact (by pressing the plate
to the specified surface)
OR Swabbing the surface and subsequently
swabbing the plate with the sample or
inserting swab into broth filled tube

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Routine Monitoring of Classified Areas

 Frequency and extent of testing is

commensurate with proximity to product and
level of risk of contamination to product
– Class 100,000 Areas
 Weekly
 Air and Selected Surfaces
– Class 10,000 Areas
 Once per use day during processing
 Air and Surface
– Class 100 Areas
 Per Process for each operating shift
 Air, Surface, Personnel
 Product Contact Surfaces (end of process)

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Utility Monitoring

 Conducted for
– Initial qualification
– Routine monitoring
– Following incursions into the utility system
 Testing is commensurate with type of system and usage
– e.g., Water for Injection (WFI) vs. Potable Water
 Water (WFI)
– Microbe, Particle, and Endotoxin Testing per use day or weekly
– Chemical/Physical test weekly
 Clean Steam
– Endotoxin and Chemical/Physical test once per month
 Compressed Gases
– Microbes and Particles
– Particles weekly and microbes monthly

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Personnel Monitoring

 Personnel greatest vehicle for

contamination into cleanrooms
 Cleanroom clothing required to
protect product from people
 Four test site locations
– Fingertips of both hands
– Forearm and Chest
 Avoid unnecessary movements
and touching of surfaces
 Follow Aseptic Technique
 Disinfect hands frequently
 Practice “First Air Rule”
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Test Processing

• Record immediate
particle air count
• Incubate microbial test
plates per procedures
• Count colonies on plate
• Identify representative
colonies
• Record all test results
1. How many CFUs?
30 2. Are they mold or bacteria?
30
 Result Analysis

Action Level: 140

Test result above maximum allowable 120

level. 100 Action Level

80

Alert Level: 60
Alert Level
Possible indication of
adverse trend in area/utility 40

performance 20

0
Test Tes t Te st Test Test Test Test

Passing Level: #1 #2 #3 #4 #5 #6 #7

Satisfactory results

31 31
Investigations for Alert/Levels

 Regulatory Requirement
 Investigation Method
– Trend
– Describe Event
– Identify Affected Materials and Lot(s)
– Identify Root Cause/Corrective Actions
– Trend for Same Root Cause Previously
– Conclusion and Impact Statement

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Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

33 33
Environmental Monitoring Issues

 Aseptic versus Sterile Concepts

– Cleanrooms are clean to allowable levels and not sterile
– Personnel are the predominant source of cleanroom
contamination
 EM is not equivalent to a quantitative analytical test
– Accuracy is limited at low level concentrations
– Results vary with equipment used
– Adventitious contamination of the test plate occurs
– Therefore, EM levels are not product specifications
 EM most appropriate as trending tool to
demonstrate continued control and/or changes in
area performance
– Continued “snap shots” in time provide picture of overall level of
control
34 – Occasional elevated results do not indicate a loss of control34
Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

35 35
Regulations and Guidance

Increasing Requirements
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Risk Based Approach

 Regulatory Guidance often non prescriptive

for EM
– Majority of guidance addresses aseptic filling
– Identify process steps with highest risk to product
quality
– Design program to address the risk(s)
– Document rationale for program design

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 Example Regulatory Citations (483s)

 Air returns within aseptic filling rooms were obstructed by shelves,

tables, and disposal cans.
 Pressure differentials are not monitored between the filling room xx
and the adjacent construction area. There is no documentation to
show the physical integrity of the seal surrounding door between
filling room xx and the adjacent construction area.
 Procedures for the recording of pressure differentials in the
fermentation area were not followed mm/dd/yy.
 The fermentor in room XXX had a defective “leaking” valve. A pool
of water was also observed XXX tank.
 A system should be implemented in the airlock to prevent the
opening of both doors at the same time.

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Example Regulatory Citations (483s)
(Cont. )

 Qualification of the method used to detect contamination

in the fermentor was incomplete in that raw data did not
include counts of the test organisms.
 Aseptic personnel practices observed during the fill of
XXXXX on mm/dd/yy were inappropriate in that operators
did not routinely disinfect gloves between manufacturing
operations.
 The EM program does not include the use of microbial
growth media that is optimum for the propagation of yeast
or mold.
 The rationale and justification of environmental monitoring
samples for all locations is not documented.
 The firm has not conducted recovery studies to qualify the
….method and material utilized…..

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Presentation Outline

 What is contamination?
 How is contamination controlled?
 Environmental Monitoring Program
 Environmental Monitoring Issues
 Regulatory Overview and Citations
 References and Reading Suggestions

40 40
References and Reading Suggestions

 Title 21 Code of Federal Regulations (CFR) Parts 210, 211 and

600
 FDA 2004 Guideline on Sterile Drug Products Produced by
Aseptic Processing
 Q7A GMP Guidance for API
 Rules Governing Medicinal Products in the European Union
Volume IV – ‘Good Manufacturing Practice for Medicinal
Products’ (EudraLex) –
– Annex 1 ‘Manufacture of Sterile Medicinal Products’
– Annex 2 Manufacture of Biological Medicinal Products
– Annex 18 GMP for APIs
 US Pharmacopoeia
– General Chapter <1116> Microbiological Evaluation of Cleanrooms and
Other Controlled Environments
 ISPE
– Baseline Pharmaceutical Engineering Guide, A Guide for New Facilities.
Volume 6 Pharmaceuticals, 2004.
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References and Reading Suggestions
(Cont.)

 International Standards Organization (ISO)

– Cleanrooms and Associated Controlled Environments – Part 1: 14644-
1: Classification of Air Cleanliness1.
– Cleanrooms and Associated Controlled Environments – Part 2: 14644-
2: Specification for Testing and Monitoring to Prove Continued
Compliance with ISO 14644-1.
 PDA
– Technical Report (TR) 13 Fundamentals of Environmental Monitoring
Program
– TR 22 Process Simulation Testing for Aseptically Filled Products
– TR 28 Process simulation Testing for Sterile Bulk Pharmaceuticals
 Books
– Microbiology in Pharmaceutical Manufacturing. Editor R. Prince, PDA
and Davis Horwood International Publishing, Ltd. 2001.

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