PhEn-602

Notes # 6
J. Manfredi
Pharmaceutical Facility Design
Spring 2009 1
Unidirectional Flow Clean Rooms
& Air Velocity
‹ Parenteral Drug Association –
guidance on air velocity
‹ Topic A: Airflow Velocity How often tested?
‹ Problem Statement: When do velocity
measurements have to be taken?
‹ Recommendation
Airflow velocity measurements should be taken
during operational and performance qualification
studies. Frequency of routine monitoring should
be at least annually. HEPA filters in critical areas
should be tested semi-annually. More frequent
measurements may be appropriate if other
measures of clean room quality indicate a
significant deviation.
Pharmaceutical Facility Design
Spring 2009 2
Unidirectional Flow Clean Rooms
Velocity Testing
‹ Airflow pattern studies should be repeated when any
changes are made that might have an impact on the
velocity measurements outside validated acceptance
criteria (i.e., changes to air handling systems, aseptic
processing equipment, HEPA filters). Evaluation of such
impact should be made following applicable change
management procedures. Airflow measurements can be
area or line-specific.
‹ Rationale for Recommendation
Airflow velocity is measured to ensure adequate airflow
to protect exposed product, product contact packaging
components, and product contact surfaces. It is also
measured to ensure there have been no significant
changes to the HVAC system. Airflow criteria are
established during qualification studies.
Pharmaceutical Facility Design
Spring 2009 3
Unidirectional Flow Clean Rooms
Velocity Testing
‹ During qualification, airflow pattern
tests/smoke studies should be
performed to establish the
acceptable velocity range.
• Where is velocity measured?
‹ Filter face?
‹ 6” down?

‹ Work level?

• For homework
Pharmaceutical Facility Design
Spring 2009 4
 Clean Rooms and Controlled
Environments
‹ FDA Aseptic guidelines: section 4, Buildings and
Facilities: “Clean area control parameters should be
supported by microbiological and particle data obtained
during qualification studies. Initial cleanroom
qualification includes, in part, an assessment of air
quality under as-built, static conditions. It is important for
area qualification and classification to place most
emphasis on data generated under dynamic conditions
(i.e., with personnel present, equipment in place, and
operations ongoing). An adequate aseptic processing
facility monitoring program also will assess conformance
with specified clean area classifications under dynamic
conditions on a routine basis”.
Pharmaceutical Facility Design
Spring 2009 5
 FDA Aseptic Guidelines
Class of Clean Room
‹ Supporting Clean Areas
‹ Supporting clean areas can have various classifications
and functions. Many support areas function as zones in
which nonsterile components, formulated products, in-
process materials, equipment, and container/closures
are prepared, held, or transferred. These environments
are soundly designed when they minimize the level of
particle contaminants in the final product and control the
microbiological content (bioburden) of articles and
components that are subsequently sterilized.

Pharmaceutical Facility Design

Spring 2009 6
 FDA Aseptic Guidelines
Class of Clean Room
‹ Supporting Clean Areas
‹ The nature of the activities conducted in a supporting
clean area determines its classification. FDA
recommends that the area immediately adjacent to
the aseptic processing line meet, at a minimum,
Class 10,000 (ISO 7) standards (see Table 1) under
dynamic conditions. Manufacturers can also classify
this area as Class 1,000 (ISO 6) or maintain the entire
aseptic filling room at Class 100 (ISO 5). An area
classified at a Class 100,000 (ISO 8) air cleanliness level
is appropriate for less critical activities (e.g., equipment
cleaning).
Pharmaceutical Facility Design
Spring 2009 7
 FDA Aseptic Guidelines
Class of Clean Room
FDA Aseptic Guidelines from 1987
Controlled Area:

“A controlled area is one in which unsterilized drug

product, in-process materials or containers/closures are
prepared.…. acceptable air quality if it has a per-cubic-
foot particle count of not more than 100,000 in a size
range of 0.5 micron and larger (Class 100,000) when
measured in the vicinity of the exposed articles during
periods of activity.

Pharmaceutical Facility Design

Spring 2009 8
 Clean Rooms and Controlled
Environments
‹ Pharmaceutical:
• A controlled* area where personnel are required to be in
a minimal amount of gowning. E.g.: Packing hall.
Typical gowning consists of coat, hat and shoe covers

*controlled term is used generically. It is different from the

controlled area referenced in the 1987 Aseptic guidelines.

‹ Pharmaceutical with local monitoring:

• A pharmaceutical area that has at least some portions
designed as Class 100,000 at rest.

Pharmaceutical Facility Design

Spring 2009 9
Clean Rooms and Controlled
Environments

‹ Pharmaceutical also called

“Controlled Not Classified”
• Often designed as EU Grade D

Pharmaceutical Facility Design

Spring 2009 10
Microbiological Considerations
&
Environmental Monitoring

Pharmaceutical Facility Design

Spring 2009 11
 Lesson Objectives:
‹ Discuss sources of micro-contaminants
‹ Discuss regulatory requirements
‹ Identify types of microbiological
sampling in the clean room
‹ Discuss sampling equipment used
‹ Discuss environmental monitoring
program
Pharmaceutical Facility Design
Spring 2009 12
 Reference Documents
‹ USP, <1116>, “Microbiological
Evaluation of Clean Rooms and Other
Controlled Environments”
‹ FDA, 1987. Guideline on Sterile Drug
Products Produced by Aseptic
Processing.
‹ FDA, 2002. Draft Guideline on Sterile
Drug Products Produced by Aseptic
Processing
‹ European Commission Good
Manufacturing Practices: Medicinal
Products for Human and Veterinary Use
Pharmaceutical Facility Design
Spring 2009 13
Microbiological Considerations
‹ FDA Aseptic Guidelines: first
document to establish general
micro limits
‹ USP and EU GMP’s provide more
specific limits for micro-
contaminants in the Clean Room
in three areas:
• Airborne
• Surface
• Personnel
Pharmaceutical Facility Design
Spring 2009 14
Microbiological Considerations
‹ People are often the only
source of micro-organisms
or viable particulates in
the clean room
‹ Micro-organisms are
continually dispersed from
people in the room
‹ Testing for
microorganisms in the “as-
built” or “at rest” state is
of little value. Pharmaceutical Facility Design
Spring 2009 15
Microbiological Considerations
‹ Other sources of microbial
contamination:
• Water
• Gases
• Raw materials
‹ Important to monitor micro levels
in these systems as well as air
• Will not be discussed in PhEn-602

Pharmaceutical Facility Design

Spring 2009 16
Microbiological Considerations

‹ Unit of measure is colony forming

unit (cfu) (Viability?)
‹ FS 209E and ISO 14644-1 do not

contain any specific limits for

microbial contaminants
‹ ISO: Developing new standards for

Micro limits in clean rooms

Pharmaceutical Facility Design

Spring 2009 17
 Critical Areas
‹ Critical Areas (sites, zones, surfaces)
are identified as those areas where
sterilized product or container and/or
closures are exposed to the
environment
• Critical sites, zones, surfaces should be
monitored most rigorously
• Class 100 areas (EU Grade A)
• Organisms recovered from critical areas
should be identified to genus and species
for possible investigation
Pharmaceutical Facility Design
Spring 2009 18
 Critical Area Examples:
‹ Room air in areas with product or
container exposure
• Path of any open containers
‹ Manufacturing equipment surfaces
• Component loading areas
• Filling Stations
‹ Storage containers
‹ Gloved hands
‹ Aseptic connections
Pharmaceutical Facility Design
Spring 2009 19
Non-Critical Area Examples:

‹ Areas that do not come in contact

with the sterilized product or
container/closures.
• Class 10,000 and 100,000 (C, D)
• Monitoring becomes less rigorous as
the classification number increases

Pharmaceutical Facility Design

Spring 2009 20
Non-Critical Area Example

‹ Class 100,000 Areas

• “Non-sterile” portion of changing
areas. Example: washrooms
• Preparation of bulk solutions prior to
sterile filtration
• Preparation of components prior to
sterilization

Pharmaceutical Facility Design

Spring 2009 21
Microbiological Considerations
1987 FDA Aseptic Guideline Limits:
‹ Controlled areas: (Class 100,000 areas)
• “with regard to microbial quality, an
incidence of no more than 25 colony
forming units per 10 cubic feet is
acceptable.”
‹ Critical areas: (Class 100 areas)
• “air should also be of a high microbial
quality….no more than 1 colony
forming unit per 10 cubic feet is
considered attainable and acceptable”.
Pharmaceutical Facility Design
Spring 2009 22
FDA Aseptic Guidelines: Clean Room Class
TABLE 1 - Air
Classifications CDER Aseptic Guidelines - 2004
Microbiological
Settling Plates
Clean Area G.T. or equal to Microbiological Action Levels
Classification (0.5 ISO 0.5 micron Active Air Action (diam. 90mm,
3 3 3
micron particles/ft ) Designation (particles/m ) Levels (cfu/m ) cfu/4 hours)

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
Pharmaceutical Facility Design
Spring 2009 23
Microbiological Considerations
‹ EU Limits for Microbial Contaminants
• Differs for Grades A, B, C, D
• No requirements for the “at rest” state
• Requirements for
‹ Airborne

‹ Surfaces

‹ Personnel Gowns

‹ Personnel Gloves

• EU has limits for surfaces contained in

Class 100,000 areas. FDA & USP do not
Pharmaceutical Facility Design
Spring 2009 24
Microbiological Considerations
‹ EU Limits for Microbial Contaminants
• Airborne:
‹ Grade A: less than 1 cfu/cubic

meter
‹ Grade B: less than 10 cfu/cubic
meter
‹ Grade C: less than 100 cfu/cubic
meter
‹ Grade D: less than 200 cfu/cubic
meter
Pharmaceutical Facility Design
Spring 2009 25
 Microbiological Considerations

‹ USP Limits for Microbial Contaminants

• Controlled areas: (Class 100,000 areas)
• Class 10,000 Areas (sometimes referred
to as “sub-critical” areas)
• Critical areas: (Class 100 areas)

Pharmaceutical Facility Design

Spring 2009 26
Microbiological Considerations

‹ USP Limits for Microbial Contaminants

• Requirements for
‹ Airborne

‹ Surfaces

‹ Personnel Gowns

‹ Personnel Gloves

Pharmaceutical Facility Design

Spring 2009 27
USP Airborne Viable Limits
Room Classification cfu/m3 cfu/ft3

Class 100 <3 <0.1

Class 10,000 <20 <0.5

Class 100,000 <100 <2.5

Note: These limits are consistent with the 1987 FDA

guidelines for Class 100, and Class 100,000 areas
Pharmaceutical Facility Design
Spring 2009 28
USP Surface
Limits
Class cfu/contact plate (24-30cm2)

100 3 (including floor)

10,000 5
10 (floor)
Pharmaceutical Facility Design
Spring 2009 29
 USP Limits for Personnel

Class cfu/contact plate cfu/contact plate

Gloves Garb

100 3 5

10,000 10 20

Pharmaceutical Facility Design

Spring 2009 30
 USP Limits for Personnel

‹ No limits for personnel in Class

100,000 areas
‹ Microbial monitoring of personnel

is not required for Class 100,000

areas

Pharmaceutical Facility Design

Spring 2009 31
 Environmental Monitoring
‹ Environmental Control:
Steps taken through facility design and
construction, testing and validation, personnel
practices and cleaning and sanitization to limit
micro-organisms in the clean room.

‹ Environmental Monitoring:
Routine microbiological monitoring provides a
series of snapshots of the microbiological profile
of a controlled environment. Routine monitoring
ensures that systems continue to provide an
environment of consistent quality.

Pharmaceutical Facility Design

Spring 2009 32
 Environmental Monitoring
‹ EM Objectives
• Monitor the effectiveness of the cleaning
and sanitization
• Monitor effectiveness of gowning and
training of personnel in aseptic areas
• Provide information for trending and to
identify excursions from normal operating
parameters
• Verify that we continue to maintain an
environmentally controlled system as
initially established in the validation of the
clean room
Pharmaceutical Facility Design
Spring 2009 33
Environmental Monitoring
‹ Agar:
• Jelly-type material
• Nutrients added to support
microbiological growth

Agar

Pharmaceutical Facility Design

Spring 2009 34
 Environmental Monitoring

‹ Airborne Viables are measured:

• Volumetric (i.e. cfu/m3)
• Settling plate (i.e. measured in cfu per
unit time collected)
‹ 1 cfu/4 hr. period, for a 90 mm settling
plate, is requirement for a Grade A room).

Pharmaceutical Facility Design

Spring 2009 35
 Environmental Monitoring

‹ Airborne Viables
• Volumetric
sampling typically
done by sampling
a specific volume
of air per unit
time.
• Slit to Agar (STA)
sampler typically
used
Pharmaceutical Facility Design
Spring 2009 36
 Environmental Monitoring
AIR FROM CLEAN
ROOM
STA Machine
Revolving agar plate

SLIT

AGAR PLATE

Pharmaceutical Facility Design

Spring 2009 37
 Environmental Monitoring
‹ Devices Used for Monitoring Air:

Pharmaceutical Facility Design

Spring 2009 38
 Environmental Monitoring
‹ Air through STA slit travels at a
high velocity
• Forced to turn 90 degree so particles
leave the air stream and impact the
agar surface
• When incubated at a suitable
temperature, for specific period of
time, microbe carrying particles will
grow to form a colony
• Count the colonies to obtain a count.
Pharmaceutical Facility Design
Spring 2009 39
 Environmental Monitoring

‹ STA samples typically 30–180

liters per minute approximately
• We can determine how many liters of
air are sampled.
• Knowing the counts and the volume,
we can obtain the cfu/m3

Pharmaceutical Facility Design

Spring 2009 40
 Slit to Agar (STA) Sampler
‹ Change out plate every 60 minutes
• Microbiologist gowns and enters fill room every hour
‹ Colonies are “seen” as dots in the contact plate
• Different colors, clear or translucent
‹ Incubate at different temperatures
• First two days at 30 - 35 C
• Next five days at 20 - 25 C
• Total of five to seven days for incubation
‹ Various types of spores may be present
‹ Mold, if present will grow in a large formation
covering most of the plate.
• Individual colonies would lie underneath the mold
‹ reverse the plate to see them.
Pharmaceutical Facility Design
Spring 2009 41
 Environmental Monitoring

‹ Airborne Viable- Settling plate

• Petri dish left in room for a specific
period of time.
• Can correlate number of micro-
organisms deposited onto the settling
plate to # particles deposited onto an
open product container
‹ Proportional areas.

Pharmaceutical Facility Design

Spring 2009 42
 Environmental Monitoring
‹ Surface Viables: Measured in cfu
per contact plate.
• Contact Plates often used: RODAC
(Replicate Organisms Detection and
Counting)
• Agar is pushed onto the clean room
surface to be sampled
• Then incubate to obtain count per
plate
Pharmaceutical Facility Design
Spring 2009 43
 Environmental Monitoring

‹ RODAC plates are appropriate for

flat surfaces
• Can obtain CFU per plate, or CFU per
unit area (per sq inch)

Pharmaceutical Facility Design

Spring 2009 44
 Environmental Monitoring
Other methods of surface sampling
including uneven surfaces:
‹ Cotton Swabs

• Rub surface to be sampled, then pass over

an agar plate.
• Plate then incubated and counted
‹ Contact Strips & Slides
• Strips are removed from containers
• Applied to the surface to be sampled
• Incubate, then count
Pharmaceutical Facility Design
Spring 2009 45
 Environmental Monitoring
Sampling Personnel
‹ Sample gloves – Fingers tips pressed
against an agar plate
‹ Garments/gowns: Press plate or
contact strip against the clothing. Best
done as they exit the clean room.
• Could sample at various locations on the
body
‹ Initial qualification sampling more in-
depth than routine sampling
Pharmaceutical Facility Design
Spring 2009 46
 EM & FDA Expectations
‹ FDA expects firms to have a thorough
Environmental Monitoring program
• For critical areas, clear action and alert limits
must be specified
• SOP’s must establish frequency of sampling,
and type of sampling to be performed, for
each type of clean room
• Trending must be performed
• Deviations from normal results must be
documented on an investigation report
• Excursions must be explained
• Corrective action must be taken if applicable
Pharmaceutical Facility Design
Spring 2009 47
 Environmental Monitoring
Environmental Control vs. Monitoring

Environmental Control
Steps taken in the facility/clean room
design construction, operation, personnel
behaviors, and cleaning and sanitization
to limit the presence of micro-organisms
in the clean room environment.

Pharmaceutical Facility Design

Spring 2009 48
Environmental Control Requires:
‹ Providing/maintaining:
• Appropriate sterile air flow and air
changes
• Effective sanitization and disinfection
• Appropriate controls of temperature
and relative humidity (RH)
• Equipment cleaning and sterilizing
‹ Especially important
• Appropriate training and re-training of
operators/personnel
Pharmaceutical Facility Design
Spring 2009 49
 Environmental Monitoring
Environmental Control vs. Monitoring

Environmental Monitoring:
Routine microbiological monitoring
program that provides a series of snapshots
of the microbiological profile of a controlled
environment. Routine monitoring ensures
that systems continue to provide an
environment of consistent quality.
Pharmaceutical Facility Design
Spring 2009 50
 “Alert” Level
‹ Warning of a potential problem
‹ Exceeding the alert level signals a
potential drift from normal operating
conditions
• Should define the alert level based on history
• Excursions may or may not require action, but
they do require that the situation be closely
monitored
• Frequent alert level excursions will require
action

Pharmaceutical Facility Design

Spring 2009 51
 “Action” Level
‹ Exceeding the action level signals a drift
from normal operating conditions
• Excursions beyond the action level will
require some type of investigation and/or
action
• The investigation will result in some sort of
action
• The action will depend on the frequency of
the excursion, the usage of the room, the
activity in the room, etc.

Pharmaceutical Facility Design

Spring 2009 52
 Environmental Monitoring
‹ There should be a “cushion” between
the design and operating level of the
clean room, and the alert and action
levels.
‹ There should be a “cushion” between
the alert level and action level.
‹ For less critical areas, such as
controlled areas, alert and action levels
may be equivalent

Pharmaceutical Facility Design

Spring 2009 53
“Alert” and “Action” Levels

Alert Range

Design Range
Normal Operating Range

Action Range

Pharmaceutical Facility Design

Spring 2009 54
Example: “Alert” and “Action” Levels

Pharmaceutical Facility Design

Spring 2009 55
Example: Classification Levels within an
Aseptic Facility
LOCKER
ROOM -
ENTRY
AREA

PHARMACEUTICAL
AREA

CLASS 100,000 CLASS 100,000

CORRIDOR PREPARATION AREA

ASEPTIC CORRIDOR
CLASS 10,000

CLASS 10,000
BACKGROUND

FILLING
ROOM CLASS
100
REGION

CLASS 100,000
CORRIDOR

PHARMACEUTICAL
AREA

Pharmaceutical Facility Design

Spring 2009 56