FDA’s GMP Inspection with

special regard to Aseptic

Process for Drugs & Biologicals

Professor Chung Keel Lee, Ph.D.

Special Advisor to the Minister, MFDS

Invited Professor, EWU & SNU
Advisor to World Health Organization(WHO)
Chair, Korea Affiliate, International Society for
Pharmaceutical Engineering(ISPE)

March 22, 2018

Gyeongin Regional Office of MFDS
The countries visited by CKL as an inspector and/or
trainer since 1987

Argentina Chile Denmark Korea Myanmar Taiwan Venezuela

Australia China Indonesia Malaysia Philippines Thailand Vietnam
Brazil Colombia Iran Mexico Russia USA
Bulgaria Cuba Japan Mongolia Singapore 2
Uzbekistan
 Contents
• Four Basic Elements of CGMP : “4Ms”
• System-based Inspection
• Latest GMP Trends(QbD)
• Sterile Drug Products Produced by Aseptic
Processing
• Significant Deficiencies by System-based
Inspection
• Question & Answer Session
Four Basic Elements of CGMP
 “4Ms”

Qualification Buildings
Adequate training Facilities
Equipment
Tools
Men Machinery
“4Ms”
Products Materials Methods
Reagents Manufacturing
Components Control
Containers & Closures Validation
Labels Documentation
Four Basic Elements of CGMP
 Men

• Organization : Independence
• Qualification
• Training
• Personnel Responsibilities
 Four Basic Elements of CGMP
 Materials
• Receipt Untested Components,
• Sampling Drug product Containers
• & Closures
Testing
• Release
• Retesting
• Cell Banking
• Viral Seeds
• Reagents
• Labels
• Products
 Four Basic Elements of CGMP
 Machinery
• Building Design HEPA
• HVAC System filter
• Water For Injection
System
• Clean Steam System
• Washing & Toilet
Facilities
• Laminar Flow Hoods.
Comparison of Air Cleanliness Classifications
Descriptive Class 100 Class 10,000 Class 100,000 ND

≥ 0.5 ㎛
100 10,000 100,000 ND
FDA /ft3
In Operation Action
Level 1 10 100 ND
CFU/m3
Descriptive A B C D ND
≥ 0.5 ㎛
3,520 3,520 352,000 3,520,000
/m3
At Rest
≥5㎛
EU, 20 29 2,900 29,000
/m3
WHO
, ≥ 0.5 ㎛
3,520 352,000 3,520,000 ND
PIC/S /m3

In Operation ≥5㎛
20 2,900 29,000 ND
/m3

CFU/m3 <1 < 10 < 100 < 200

ISPE Descriptive Grade 5 Grade 7 Grade 8 CNC+ CNC

Descriptive ISO. 5 ISO. 7 ISO. 8 ISO. 9

≥ 0.5 ㎛
ISO 3,520 352,000 3,520,000 35,200,000
/m3
In Operation
≥5㎛
20 2,930 29,300 293,000
/m3
 3 Occupancy States

 As-Built condition
- Where the installation is complete with all services
connected and functioning but with no equipment
& personnel present.
 At-Rest condition
- Where the installation is complete with equipment
installed and operating but with no personnel
present.
 Operational condition
- Where the installation is functioning with the
specified No. of personnel present & equipment
operating.
 Four Basic Elements of CGMP
 Methods
• Production
• Sampling & Testing
• Environmental Monitoring
• Packaging & Labeling
• Validation
• Documentation Drug Products
• Storage - Under appropriate conditions
Quarantine

Quality Control(QC)

- The oldest stock product

Release is distributed first
- A system to readily determine
the distribution of drug
 System Based Inspection

 Quality System
 Facility & Equipment
 Materials
 Production
 Laboratory Controls
 Packaging & Labeling
 CGMP vs. Systems

CGMP Subpart Qual. F/E Mat. Prod PKG/L Lab.

B. Organization & Personnel      
C. Buildings & Facilities 
D. Equipment 
E. Control of Components & Drug
Product  
containers & closures

F. Production & Process Controls  

G. Packaging & Labeling Control  
H. Holding & Distribution 
I. Laboratory Controls  
J. Records & Reports      
K. Returned & Salvaged Drug Products  
 Latest GMP Trends(QbD)
 Introduction
 2002 : Pharmaceutical CGMP Initiative for
the 21st Century-A Risk Based Approach

The Pharmaceutical Quality for the

21st Century-A Risk Based Approach
 Latest GMP Trends(QbD)
 Traditional vs. New Enhanced Approach

Research & Development

Clinical Commercial Post-market
Pre-clinical Manufacturing Surveillance
Ⅰ Ⅱ Ⅲ

Traditional

Risk-based Approach
Enhanced
Quality System Approach

The entire lifecycle of a product

 Latest GMP Trends(QbD)
 Ishikawa (Fishbone) Diagram (risk assessment tool)

 QTPP
 CQA of the product
 Input variables : materials etc.
 Process parameters : temp., time, humidity etc.
 Multidimensional combination & interaction → Design Space
 Real time release (test)
 Quality by Design → Regulatory flexibility
 Latest GMP Trends
 US FDA’s Guidances with Enhanced Approach (QbD)
 Sterile Drug Products Produced by Aseptic Processing –CGMP
(Sept., 2004)
 Process Analytical Technology (PAT) : A Framework for
Innovative Pharmaceutical Development, Manufacturing &
Quality Assurance (Sept., 2004)
 Quality Systems Approach to Pharmaceutical CGMP Regulations
(Sept., 2006)
 Process Validation : General Principles & Practices (Jan., 2011)
 Latest GMP Trends
 ICH Harmonized Tripartite Guidelines

 Pharmaceutical Development (ICH Q8:R2) (Aug., 2009)

 Quality Risk Management (ICH Q9) (Nov., 2005)

 Pharmaceutical Quality System (ICH Q10) (June, 2008)

 Development & Manufacture of Drug Substances (ICH Q11)

(Nov., 2012)
 Sterile Drug Products produced
by Aseptic Processing
I. Introduction VIII. Time Limitations
II. Background IX. Validation of Aseptic
III. Scope Processing & Sterilization
IV. Building & Facilities X. Laboratory Controls
V. Personnel Training XI. Sterility Testing
VI. Components & XII. Batch Record Review
Containers/Closures
VII. Endotoxin Control
 I. Introduction
• Replacement of 1987 guideline on sterile
drug products produced by aseptic
processing (aseptic processing guideline)
• When manufacturing sterile drug and
biological products using aseptic
processing
 II . Background
A. Regulatory Framework
 Where it is impossible to comply with the
applicable regulations in both parts 600
through 680 and parts 210 & 211, the
regulation specifically applicable to the drug
product in question shall supercede the
more general regulations.
 III. Scope
• This guidance updates the 1987 Aseptic
Processing Guideline with respect to:
– personnel qualification
– cleanroom design
– process design
– quality control
– environmental monitoring
– review of production records
 IV. Buildings and Facilities
• Clean area air classifications: measured
at work level under dynamic conditions
Clean Area *CFU/4-hr
Classificatio ISO > 0.5 m *CFU/m 3 (passive S.
n Designatio particles/ (active S.) diam. 90mm)
(≥ 0.5 m n m3
particles/ft 3)

100 5 3,520 <1** <1

1000 6 35,200 <7 <3

10,000 7 352,000 <10 <5

100,000 8 3,520,000 <100 <50

* You may establish alternate levels due to the nature of the operation or method of analysis.
** Normally yield no microbiological contaminants.
 IV. Buildings and Facilities
A. Critical Area-Class 100 (ISO 5)
 A critical area is one in which the sterilized
drug product, containers, and closures are
exposed to environmental conditions that
must be designed to maintain product sterility.
 Particles:
– Extraneous contaminant themselves
– Act as a vehicle for microorganisms.
– To be measured not more than 1 foot away from
the work site.
– Regular monitoring should be performed during
each production shift.
 IV. Buildings and Facilities
A. Critical Area-Class 100 (ISO 5)
 High levels of product particles
– Qualification of the area under dynamic conditions
without actual filling function
 In situ air pattern under dynamic conditions
– Unidirectional air flow
– Sweeping action over & away from the product
• A velocity of 0.45 meters/second (90
feet/minute) ±20%
 IV. Buildings and Facilities
B. Supporting Clean Areas
 To be designed to minimize the level of particle
contaminants & microbiological content (bioburden)
 The area immediately adjacent to the aseptic
processing line should meet, at a minimum, class
10,000 (ISO 7) standards under dynamic conditions.
Class 1,000 (ISO 6) or Class 100 may apply to this
area.
 Class 100,000 (ISO 8) air cleanliness level is
appropriate for less critical activities (e.g.,
equipment cleaning).
 IV. Buildings and Facilities
C. Clean Area Separation
 Airflow from higher cleanliness area to
adjacent less clean areas.
 A substantial positive pressure differentials
for rooms of higher air cleanliness
 10-15 Pa (0.04-0.06 inches of water gauge )
maintained between rooms of differing
classification with doors closed.
 IV. Buildings and Facilities
C. Clean Area Separation
 At least 12.5 Pa should be maintained
between the aseptic processing room and
unclassified room adjacent to the aseptic
processing room.
 Pressure differentials should be monitored
continuously & frequently recorded.
 IV. Buildings and Facilities
C. Clean Area Separation
• For class 100,000 (ISO 8) at least 20 air
changes per hour is acceptable.
Significantly higher air change rates are
normally needed for class 10,000 & Class
100 areas.
• A suitable facility monitoring system (i.e.,
alarms) is needed.
 IV. Buildings and Facilities
D. Air Filtration
1. Membrane
 Membrane filters (sterile) can be used for
compressed gas, autoclave air lines, lyophilizer
vacuum breaks, tanks containing sterilized materials.
 Gas filters (including vent filters) should be dry.
 Filters that supply sterile gases should be integrity
tested upon installation & periodically thereafter
(e.g., end of use).
 IV. Buildings and Facilities
D. Air Filtration
2. HEPA
• Leak testing at installation
• 2 x / year for aseptic processing room
• Facility renovation, media fill failure, drug product
sterility failure... : additional testing
• Leak test on dry heat depyrogenation tunnels &
ovens(e.g. glass vials)
 IV. Buildings and Facilities
D. Air Filtration
2. HEPA
• DOP (DiOctylPhthalate) or PAO (Poly-Alpha-Olefin)
– Leak testing aerosols
• Alternative aerosols
– Not promoting microbial growth
• Filter efficiency test
– Monodispersed aerosol of 0.3μ particles
– Retaining at least 99.97%
IV. Buildings and Facilities

D. Air Filtration
2. HEPA
 Filter Integrity test
– Polydispersed aerosol (mean < 1μ but > 0.3 μ)
– Sufficient No. of particles at ~0.3μ.
– Test in place
– Scan at 1~2 inches from the filter face
– Sampling rate at ≥1 ft3/min.
– > 99.99% retention or < 0.01% leak
• Uniformity of velocity across the filter
– Variations in velocity can cause turbulence.
– Velocity of unidirectional air
» At 6 inches from the filter face and
» At work level in critical area
 IV. Buildings and Facilities
E. Design
• Aseptic processes designed to minimize
exposure of sterile articles to contamination
hazards
• Flow of personnel, materials, products,
equipment, air & waste
• Minimum No. of personnel in aseptic
processing area
• SIP (sterilize-in-place) or automation of
processes can reduce risk to the product.
 IV. Buildings and Facilities
E. Design
• A partially closed sterile product should be
transferred only in critical areas.
– The area between a filling line & the lyophilizer
• Appropriately designed transfer equipment can be
qualified for this purpose.
• Airlocks with interlocking doors between aseptic
processing area entrance and unclassified area.
• Stoppered vials should be under protection until
completion of the crimping step.
 IV. Buildings and Facilities
E. Design
• Seamless & rounded floor to wall junctions,
readily accessible corners…
• Sanitary fittings & valves for processing
equipment
• No drains in aseptic processing areas
• Avoid horizonal surfaces or ledges
• SOPs on returning a facility to operating
conditions following a shutdown
 V. Personnel Training
A. Personnel
• Evaluate each operator’s conformance to
written procedures during operation.
• QC’s regular oversight on adherence to
established, written procedures during
manufacturing
• Techniques to maintain sterility of sterile
items & surfaces:
– Contact sterile materials only with sterile
instruments.
– Move slowly and deliberately.
 V. Personnel Training
A. Personnel
• Techniques to maintain sterility of sterile
items & surfaces:
– Keep the entire body out of the path of
unidirectional airflow.
– Approach from the side & not above the
production in vertical laminar flow.
– Refrain from speaking when in direct proximity to
the critical area.
– Maintain proper gown control.
 V. Personnel Training
A. Personnel
 Gowns: sterile, nonshedding, covering the skin &
hair
– Common elements of gowns: face-masks, hoods,
beard/moustache covers, protective goggles
– Adequate barrier should be created by the
overlapping of gown components.
– Gloves should be sanitized frequently.
• Microbiological surface sampling of several
locations on gown : e.g., glove fingers, facemask,
forearms, chest etc.
 V. Personnel Training
A. Personnel
• Periodic requalification following an initial
assessment of gowning
• Annual requalification for automated
operations
B. Laboratory Personnel
• The basic principles of training, aseptic
technique, and personnel qualification in
aseptic manufacturing also are applicable to
those performing aseptic sampling and
microbiological laboratory analyses.
 V. Personnel Training
C. Monitoring Program
 Obtain surface samples of each operator's gown
including gloves & other locations of the gown on a
daily basis, or in association with each lot.
 Asepsis is fundamental to an aseptic processing
operation.
 Sanitizing gloves just prior to sampling is
inappropriate because it can prevent recovery of
microorganisms that were present during an aseptic
manipulation.
 VI. Components and
Container/Closures
A. Components
 Active ingredients, water for injection (WFI),
and other excipients: acceptable limits of
microbial content (e.g., bioburden, endotoxin)
should be established.
 Sterile-filtration of a solution formed by
dissolving the component (s) in a solvent such
as WFI, USP, is used when the solution is
adversely affected by heat.
 VI. Components and
Container/Closures
A. Components
 Dry heat sterilization is good for components
that are heat stable and insoluble. Heat
penetration & distribution studies are
needed for powder sterilization because of
the insulating effects of the powder.
 Irradiation can be used to sterilize some
components.
 VI. Components and
Container/Closures
B. Containers/Closures
1. Preparation
• Containers & closures should be sterile and, for
parenteral drug nonpyrogenic.
• Time limits for holding sterile & depyrogenated
containers & closures
• Final rinse water should be purified water and, for
parenteral products WFI, USP.
• Subjecting glass containers to dry heat
accomplishes both sterilization & depyrogenation.
 VI. Components and
Container/Closures
B. Containers/Closures
1. Preparation
• Ethylene oxide (EtO) : effective surface sterilant
– Temperature, pressure, humidity, gas
concentration, exposure time, degassing,
aeration, & residuals should be specified &
monitored.
– Biological indicators are of special importance in
demonstrating the effectiveness of EtO & other
gas sterilization processes.
• Rubber closures (e.g., stoppers and syringe
plungers) :
– Multiple cycles of washing / rinsing prior to sterilization
 VI. Components and
Container/Closures
B. Containers/Closures
1. Preparation
• Rubber closures (e.g., stoppers and syringe plungers) :
– Initial rinses with PW, USP followed by
– Final rinse with WFI, USP for parenteral products
– Depyrogenation by multiple rinses of hot WFI
– The time between washing / drying and sterilizing should be
minimized.
– Siliconization of rubber stoppers: potential sources of
contamination
• Visual identification & Certificate of Analysis review on
containers & closures may be accepted with the reliability
of the supplier’s test results established at appropriate
intervals.
 VII. Endotoxin Control
• Endotoxin contamination of an injectable
product can occur as a result of poor CGMP
controls.
• Drug product components, containers,
closures, storage time limitations, and
manufacturing equipment are among the areas
to address in establishing endotoxin control.
• Adequate cleaning, drying, and storage of
equipment will control bioburden and prevent
contribution of endotoxin load.
 VII. Endotoxin Control
• Sterilizing-grade filters and moist heat
sterilization have not been shown to be
effective in removing endotoxin.
• Some clean-in-place procedures employ initial
rinses with appropriate high purity water and/or
a cleaning agent (e.g., acid, base, surfactant),
followed by final rinses with heated WFI.
• Equipment should be dried following cleaning,
unless the equipment proceeds immediately to
the sterilization step.
 VIII. Time Limitations
• Time limits for each phase of aseptic processing
– Between the start of bulk product compounding and
its sterilization filtration
– Product exposure while on the processing line
– Storage of sterilized equipment, containers & closures
• Bioburden & endotoxin load should be assessed to
establish time limits for stages.
• The total time for product filtration should be limited
to an established maximum:
– To prevent microorganisms from penetrating the filter
– To prevent an increase in upstream bioburden &
endotoxin load
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
 An aseptic processing operation should be validated
using a microbiological growth medium in place of
the product. This ”process simulation” (media fill)
includes exposing the microbiological growth
medium to product contact surfaces of equipment,
container closure systems, critical environments,
and process manipulations to closely simulate the
same exposure that the product itself will undergo
during actual operations (e.g., start-up, sterile
ingredient additions, aseptic connections, filling,
closing).
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
• Aseptic processing validation:
– Microbial growth medium
– Process simulation (media fill)
– Exposure to product contact surfaces
– The sealed containers filled with the medium are incubated.
– Results are interpreted to assess the potential for a unit of
drug product to become contaminated during actual
operations.
– Environmental monitoring data from the process simulation
can also provide useful information for the processing line
evaluation
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
1. Study Design for Media Fill
a. Consider the following issues: simulate actual
operations + worst-case conditions
 Factors associated with the longest permitted run:
e.g., operator fatigue
 normal interventions & nonroutine interventions

(stoppages, equipment adjustments)

 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
1. Study Design for Media Fill
a. Consider the following issues: simulate actual operations
+ worst-case conditions
 Lyophilization, when applicable
 Aseptic assembly of equipment
(e.g., at start-up, during processing)
 Number of personnel and their activities
 Representative number of aseptic additions or transfers
 Shift changes, breaks, and gown changes
(when applicable)
 Type of aseptic equipment disconnections/connections
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
1. Study Design for Media Fill
a. Consider the following issues: simulate actual
operations + worst-case conditions
 Aseptic sample collections
 Line speed & configuration
 Weight checks
 Container closure systems (e.g., sizes, type)
b. A written batch record should be prepared for
each media fill run.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
2. Frequency and Number of Runs
 At least 3 consecutive separate successful runs should be
performed during initial line qualification.
 Subsequently, routine semi-annual qualification
conducted for each processing line will evaluate the state
of control of the aseptic process.
 All personnel authorized to enter the aseptic processing
room during manufacturing (i.e., technicians, maintenance
personnel…) should participate in a media fill at least
once a year.
 Facility & equipment modifications, line configuration
changes, significant changes in personnel, anomalies in
environmental testing results, container closure system
changes, extended shutdowns, or end product sterility
testing showing contaminated products may be cause for
revalidation of the system.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
3. Duration of runs
 When aseptic processing employs manual filling or
closing, or extensive manual manipulations, the
duration of the process simulation should be no less
than the length of the actual manufacturing process
to best simulate contamination risks posed by
operators.
 For lyophilization operations FDA recommends that
unsealed containers be exposed to partial
evacuation of the chamber but the vials should not
be frozen & the medium should remain in an aerobic
state.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
4. Size of runs
• 5,000 to 10,000 units
• Maximum batch size if the run size is under
5,000
• Full or close to full batch size for manually
intensive filling
• Lower number of units in case of using an
isolator due to the lack of direct human
intervention.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
5. Line Speed
 Each media fill run should evaluate a
single line speed, and the speed chosen
should be justified.
6. Environmental Condition
 Media fills should be adequately
representative of the conditions under
which actual manufacturing operations
are conducted.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
7. Media
• “soybean casein digest medium” in general
• Growth promotion test : e.g., USP indicator organisms
– Gram-positive & gram-negative bacteria
– Yeast
– Mold
• Substituted challenge : isolates from E.M. & sterility test
• <100 CFU challenge/unit
 Each unit should be filled with an appropriate quantity of
microbial growth medium to contact the inner container/
closure surfaces (the unit is inverted or swirled).
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
8. Incubation & Examination of Media-Filled
Units
• 20-30 ℃ ± 2.5℃
• Not less than 14 days
• If two temperatures are used:
– ≥ 7 days at 20-25 ℃, first
– ≥ 7 days at 30-35 ℃, second
• QC unit’s involvement
– Observation or
– Oversight throughout the examination
• All suspect units should be brought to QC
microbiologist.
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
8. Incubation & Examination of Media-Filled
Units
• Clear containers for amber or opaque containers
• All integral units → incubation
• Units not related to integrity (cosmetic defect) →
incubation
• Units that lack integrity should be rejected.
• Criteria for yield ( total units incubated / total units
filled)
 IX. Validation of Aseptic
Processing and Sterilization
A. Process Simulations
9. Interpretation of Test Results
 The process simulation run should be observed by the
QC Unit.
 Contaminated units should be reconcilable with the
approximate time and the activity being simulated during
the media fill.
 Any contaminated unit should be identified to species
level.
 Any failure investigation should assess the impact on
commercial drugs produced on the line since the last
media fill.
 Modern aseptic processing operations should normally
yield no media fill contamination.
 IX. Validation of Aseptic
Processing and Sterilization
B. Filtration Efficacy
• 0.22μ or 0.2 μ sterilization grade filters
• Sterile filtration validation:
– Microbial challenges
– Integrity test of the filters
• Brevundimonas diminuta (ATCC 19146)
• At least 107 organisms per cm2 of effective
filter area
• No passage of the challenge microorganisms
 IX. Validation of Aseptic
Processing and Sterilization
B. Filtration Efficacy
• Minimize the bioburden of the unfiltered
product.
• Direct inoculation into the drug formulation is
preferred.
• Worst-case conditions : maximum filter use
time & pressure
• Filter integrity testing : before & after use
• Forward flow and bubble point tests
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers,
and Closures
• Moist heat and dry heat sterilization
1. Qualification & Validation
• Requalification on a periodic basis
• Remove air from the autoclave chamber.
• Filter installations in piping (SIP)
– Pressure differential across the filter
– Significant temperature drop on the downstream
– Place biological indicators at downstream locations
of the filter.
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers,
and Closures
• Moist heat and dry heat sterilization
1. Qualification and Validation
• Empty chamber studies :
– Temperature & pressure mapping study with
calibrated measurement devices
– Heat penetration studies on loading configurations
– In general, the biological indicator should be
placed adjacent to the temperature sensor to
assess the correlation between microbial lethality
and predicted lethality based on thermal input.
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers,
and Closures
• Moist heat and dry heat sterilization
1. Qualification and Validation
 Articles difficult to sterilize: filters, filling manifolds,
pumps. Some other examples include certain locations
of tightly wrapped or densely packed supplies, securely
fastened load articles, lengthy tubing, the sterile filter
apparatus, hydrophobic filters, and stopper load.
 A sterility assurance level of 10-6 or better should be
demonstrated for a sterilization process.
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers,
and Closures
• Moist heat and dry heat sterilization
1. Qualification and Validation
 For more information, refer to “ Guideline for the
Submission of Documentation for Sterilization Process
Validation in Applications for Human and Veterinary
Drug Products”.
 Change control procedures should adequately address
issues such as a load configuration change or a
modification of a sterilizer.
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers, and
Closures
• Moist heat and dry heat sterilization
2. Equipment Controls & Instrument Calibration
• For both validation & routine process control, calibrate:
– Temperature & pressure monitoring devices
– Sensing devices before and after validation runs
– Devices used to monitor dwell time in the sterilizer
– D-value (resistance) of a biological indicator (e.g., spore
strips, glass ampuls) can be accepted if the reliability of a
vendor’s COA is established.
– The microbial count of a B.I.
 IX. Validation of Aseptic
Processing and Sterilization
C. Sterilization of Equipment, Containers,
and Closures
• Moist heat and dry heat sterilization
2. Equipment Controls & Instrument Calibration
 For both validation & routine process control, calibrate :
- For dry heat depyrogenation tunnels, devices used
to measure belt speed should be routinely
calibrated.
- Routine evaluation of sterilizer performance-
indicating attributes, such as equilibrium (come up)
time is important in assuring that the unit continues
to operate as per the validated conditions.
 X. Laboratory Controls
A. Environmental Monitoring
1. General Written Program
 In aseptic processing, one of the most important
laboratory controls is the environmental
monitoring program.
 Monitoring should cover all production shifts.
 Evaluate the quality of air & surfaces:
– Air, floors, walls, equipment surfaces (especially
critical surfaces)
– Critical surfaces : come in contact with the sterile
product, containers, & closures.
 X. Laboratory Controls
A. Environmental Monitoring
1. General Written Program
 Written procedures (SOPs) should include :
– Locations to be sampled
– Sample timing ; i.e., during or at the conclusion of
operations
– Sample frequency
– Duration of sampling
– Sample size : e.g., surface area, air volume
– Sampling equipment & techniques
– Alert & action levels
– Appropriate response to deviations from alert or
action levels
 X. Laboratory Controls
A. Environmental Monitoring
1. General Written Program
 Samples should be taken throughout the
classified areas of the aseptic processing facility
(e.g., aseptic corridors, gowning rooms).
 Sample sizes should be sufficient to optimize
detection of environmental contaminations at
levels that might be expected.
 Critical surface sampling should be performed at
the conclusion of the aseptic processing
operation to avoid direct contact with sterile
surfaces during processing.
 X. Laboratory Controls
A. Environmental Monitoring
2. Establishing Levels & a Trending Program
 Alert & action levels
– Based on the relationship of the sampled location to the
operation
– Based on the need to maintain adequate control
– Consider E.M.data from historical databases, media
fills, clean room qualification & sanitization studies.
 QC unit should provide routine oversight of
trends in environmental & personnel monitoring
data.
– Near-term trends: daily, weekly, monthly & quarterly
– Long-term trends: yearly, biennially,….
 X. Laboratory Controls
A. Environmental Monitoring
3. Disinfection Efficacy
 Disinfectants should be sterile, in suitable
containers (e.g., sterile) and used for no longer
than the predefined period .
 Routinely used disinfectants should be effective
against the normal microbial vegetative flora
recovered from the facility.
 Many common disinfectants are ineffective
against spores.
 Disinfection procedures should be described in
detail e.g., preparation, work sequence, contact
time etc.
 Definitions
 Sanitization : Reduces viable microorganisms to a defined
acceptable level. Normally achieved by using a chemical agent or
heat.
 Disinfection : Process by which viable microbiological agents or
eukaryotic cells are reduced to a level unlikely to produce disease
in healthy people, plants or animals.
 Decontamination : A process that reduces contaminating
substances to a defined acceptable level
 Pasteurization : The heating of milk, wines, fruit juices, etc. for
about 30 min. at 62-68℃ whereby the living bacteria are destroyed,
but the flavor or bouquet is preserved.
 Sterilization : Destroys or eliminates all viable microbes including
bacterial spores.
 Definitions (continued)
 Antiseptic : Acting against sepsis.
An antiseptic agent is one that has been
formulated for use on living
tissue(mucous membrane or skin) to
prevent or inhibit growth or action of
organisms.
 Pyrogen : A fever-producing substance called also
pyretogen, pyretic, pyrectic
Bacterial pyrogen : a fever producing
agent of bacterial origin : endotoxin
 Endotoxin : Lipopolysaccharide
Integral part of the bacterial (gram
negative) cell wall & is only released when
the integrity of the wall is disturbed.
 Depyrogenation : The removal or destruction of pyrogen
(endotoxins)
 X. Laboratory Controls
A. Environmental Monitoring
4. Monitoring Methods for Biological Quality
a. Surface monitoring
 Product contact surfaces, floors, walls & equipment
 Touch plates, swabs & contact plates
b. Active air monitoring
 Active devices : impaction, centrifugal and membrane (or
gelatin) samplers
 Quantitative air monitoring
c. Passive air monitoring (settling plates)
 Settling plates: Petri dishes containing nutrient growth
medium exposed to the environment
 Qualitative, or semi-quantitative air monitoring.
 Exposure conditions should preclude desiccation (e.g.,
caused by lengthy sampling periods and/or high airflows),
which inhibits recovery of microorganisms.
 X. Laboratory Controls
A. Environmental Monitoring
5. Recommended action level of microbiological quality (Air)

Clean Area ISO 0.5m Microbiological Microbiological

Classification particles/m3 Active Air Settling Plates
Designation Action Levels Action Levels
(0.5m (CFU/m3) (diam.
particles/ft3) 90mm:cfu/4
hrs.)

100 5 3,520 1 1
1,000 6 35,200 7 3
10.000 7 352,000 10 5
100,000 8 3,520,000 100 50

US FDA, 2004 : Sterile Drug Products Produced by Aseptic Processing-CGMP

 X. Laboratory Controls
A. Environmental Monitoring
6. Recommended action level of microbiological
quality (Surface)
Clean Area ISO 0.5m Equipment Gloves Clothing
Classification Designation particles/m3
(0.5m particles/ft3) CFU / Contact CFU / Contact CFU / Contact
Plate Plate Plate
(24-30 cm2) (24-30 cm2) (24-30 cm2)

100 5 3,520 3 3 5
(including
floor)
1,000 6 35,200 – – –

10.000 7 352,000 5 10 20
10 (floor)
100,000 8 3,520,000 – – –

USP <1116>, 2006 : Microbiological Evaluation of Clean Rooms & Other Controlled Environment
 X. Laboratory Controls
B. Microbiological Media & Identification
• Routine identification of microorganisms to the
species levels
• Culture media capable of detecting fungi (i.e.,
yeasts and molds) & bacteria
• Total aerobic bacterial count at 30~35℃ for 48~72
hours
• Total combined yeast & mold count at 20~25℃ for
5~7 days
• Growth promotion testing on all lots of prepared
media
 X. Laboratory Controls
C. Prefiltration Bioburden
• Bioburden can contribute impurities (e.g., endotoxin) to, & lead
to degradation of, the drug product.
• Minimize the bioburden in the unfiltered product.
• A prefiltration bioburden limit should be established.
D. Alternate Microbiological Test Methods :
• Rapid test methods : equivalent or better
E. Particle monitoring
• Routine particle monitoring is useful in rapidly detecting
deviations in air cleanliness.
• A result outside the established classification level at a given
location should be investigated.
 The investigation should include an evaluation of trending
data.
 XI. Sterility Testing
• Sterility testing lab. environment : comparable to
aseptic filling operations
• Use of isolator for sterility testing minimizes false
positive test result.
A. Microbiological Laboratory Controls
• Method validation : microbiological challenge
• If growth is inhibited :
– Increased dilution
– Additional membrane filter washes
– Addition of inactivating agents
• Media : sterile & growth promoting
• Personnel qualification & training
 XI. Sterility Testing
B. Sampling & Incubation
• Limited ability to detect contamination due to
small sample size (USP):
– 10,000-unit lot with 0.1 % contamination :
20 unit sample test would pass the lot with 98% chance.
– If 10% of the lot are contaminated, contamination
can be detected about 9 out of 10 cases.
 XI. Sterility Testing
B. Sampling & Incubation
 The samples should represent the entire and
processing conditions. Samples should be taken:
– At the beginning, middle, and end of the aseptic
processing operation
– In conjunction with processing interventions or
excursions
 Because of the limited sensitivity of the test, any
positive result is considered a serious CGMP issue
that should be thoroughly investigated.
 XI. Sterility Testing
C. Investigation of Sterility Positives
• A positive test would be invalid only when microbial
growth can be unequivocally ascribed to laboratory
error. When available evidence is inconclusive,
batches should be rejected.
• Investigation's persuasive evidence of the origin of
the contamination should be based on :
1. Identification (speciation) of the organism in the
sterility test
 To the species level
 Determine to see the organism in lab. & production
environment
 Advanced identification method (e.g., nucleic-acid
based) are valuable for investigation.
 When comparing the results from E.M. and sterility
positives the same methodology should be used for
identifications.
 XI. Sterility Testing
C. Investigation of Sterility Positives
 The investigation's persuasive evidence of the origin
of the contamination should be based on:
2. Record of Laboratory Tests & Deviation
3. Monitoring of Production area Environment :
 Trend analysis of microorganisms in the critical and
immediately adjacent areas
 Look at both short - and long - term environmental trend
analyses.
4. Monitoring Personnel
• Review of data & associated trends from daily monitoring
of personnel.
• Adequacy of personnel practices & training
 XI. Sterility Testing
C. Investigation of Sterility Positives
 The investigation's persuasive evidence of
the origin of the contamination should be
based on :
5. Product Presterilization Bioburden
6. Production Record Review
 Events that could have impacted on the critical zone
 The functioning of utility and/or support systems
 Whether construction or maintenance activities could
have had an adverse impact
7. Manufacturing History
 Past deviations, problems, or changes on process,
components, equipment etc.
 XII. Batch Record Review:
Process Control Documentation
• All in-process & laboratory control results
must be included in the batch production
record.
• Essential elements of the batch release
decision:
– Review of environmental & personnel monitoring
data
– Review the data relating support systems (HEPA,
HVAC, WFI, steam generator)
– Review the data on proper functioning of equipment
(e.g., batch alarms report, integrity of various filters)
 XII. Batch Record Review:
Process Control Documentation
• Interventions and/or stoppages (unplanned)
should be documented in batch records with
the associated time & duration of the event.
• Any disruption in power supply that could
affect product quality must be included in batch
production records.
Significant Deficiencies by System

A. Quality System
B. Facilities and Equipment System
C. Materials System
D. Production System
E. Packaging and Labeling System
F. Laboratory Controls System
Significant deficiencies by system

A. Quality System
1. Pattern of failure to review/approve procedures
2. Pattern of failure to document execution of operations
as required
3. Pattern of failure to review documentation
4. Pattern of failure to conduct investigations and resolve
discrepancies/failures/deviations/complaints
5. Pattern of failure to assess other systems to assure
compliance with GMP and SOPs
Significant deficiencies by system
B. Facilities and Equipment
1. Contamination with filth, objectionable microorganisms,
toxic chemicals or other drug chemicals, or a reasonable
potential for contamination, with demonstrated avenues
of contamination, such as airborne or through unclean
equipment
2. Pattern of failure to validate cleaning procedures for non-
dedicated equipment. Lack of demonstration of
effectiveness of cleaning for dedicated equipment
3. Pattern of failure to document investigation of
discrepancies
4. Pattern of failure to establish/follow a control system for
implementing changes in the equipment
5. Pattern of failure to qualify equipment, including
computers
Significant deficiencies by system
C. Materials System
1. Release of materials for use or distribution that do not
conform to established specifications
2. Pattern of failure to conduct one specific identity test for
components
3. Pattern of failure to document investigation of
discrepancies
4. Pattern of failure to establish/follow a control system for
implementing changes in the materials handling
operations
5. Lack of validation of water systems as required
depending upon the intended use of the water
6. Lack of validation of computerized processes
 Significant deficiencies by system

D. Production System
1. Pattern of failure to establish/follow a control system for
implementing changes in the production system
operations
2. Pattern of failure to document investigation of
discrepancies
3. Lack of process validation
4. Lack of validation of computerized processes
5. Pattern of incomplete or missing batch production
records
6. Pattern of nonconformance to established in-process
controls, tests, and/or specifications
Significant deficiencies by system
E. Packaging and Labeling System
1. Pattern of failure to establish/follow a control system for
implementing changes in the packaging and/or labeling
operations
2. Pattern of failure to document investigation of
discrepancies
3. Lack of validation of computerized processes
4. Lack of control of packaging and labeling operations
that may introduce a potential for mislabeling
5. Lack of packaging validation.
Significant deficiencies by system
F. Laboratory Controls System
1. Pattern of failure to establish/follow a control system for
implementing changes in the laboratory operations
2. Pattern of failure to document investigation of discrepancies
3. Lack of validation of computerized and/or automated
processes
4. Pattern of inadequate sampling practices
5. Lack of validated analytical methods
6. Pattern of failure to follow approved analytical procedures.
7. Pattern of failure to follow an adequate OOS procedure
8. Pattern of failure to retain raw data
9. Lack of stability indicating methods
10. Pattern of failure to follow stability programs