CHAPTER 16

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Airflow studies and

airflow mapping
TIM SANDLE, HEAD OF MICROBIOLOGY, BIO PRODUCTS LABORATORY, UK
MARCO BUDINI, CONSULTANT, PHARMACEUTICAL TECHNOLOGY & COMPLIANCE, ITALY
RAJESH THEMPADIYI, MANAGER, TECHNICAL TRANSFER, DR REDDY’S LABORATORIES, INDIA

16.1 Introduction
Air is of fundamental importance to cleanrooms, either as a contamination source (micro-
organisms carried in the air-stream) or as a control measure to minimise contamination (through
the supply of clean air). Therefore, controlling a cleanroom requires careful attention to air filtration
and airflow. The movement of the air in a cleanroom is an important contamination control step1.
In a cleanroom, this air movement is normally in turbulent flow (that is air moving with a non-
uniform velocity). Here, air is driven in through grills and ducts at ceiling height and removed
through low level ducts. The air in the room with its initial supply velocity is sufficient to keep it in
constant turbulence, which prevents particles and micro-organisms from settling (this is an ideal
because dead air can occur, for example, beneath tables). With clean air devices, the air is
designed to be unidirectional whereby the air direction and air velocity are designed to remove any
contamination deposited into the air-stream away from the critical area.

Understanding how air moves in a cleanroom or clean air device is critical in order to understand
how the clean air functions in relation to any contamination that might be present in the air. This is
undertaken through airflow visualisation studies (or what are sometimes called smoke studies).
The purpose of flow visualisation is to confirm the smoothness, flow patterns and other spatial and
temporal characteristics of airflow in an installation. For this, the airflow is examined through airflow
visualisation mapping whereby smoke is generated and the behaviour of the smoke studied and
then captured by a video camera.

Airflow studies can demonstrate a significant amount of information. This can relate to
contamination control in assessing whether airflows are drawing potentially contaminated air
towards a critical zone of whether certain objects in the air-stream cause contamination by forcing
the air to change direction. Careful consideration must be made of equipment and other obstacles
in the room. Where air strikes an object any contamination in the air can be deposited or where air
becomes trapped then dead air spots can develop leading to localised turbulence. It is important
that the manager of the cleanroom understands where these ‘dead spots’ can occur and then either
attempts to engineer these out or undertakes a suitable risk assessment or targets environmental
monitoring samples (viable and particulate monitoring) towards the areas of concern2.

However, it is important to note that there are no objective criteria with which to assess airflow
visualisation studies: that is the difference between a good and bad airflow study cannot be
expressed parametrically. Thus any interpretation of an airflow study rests, to a degree, on
subjectivity and when reporting airflow studies care should be taken to avoid commenting on things
that cannot be clearly seen by the smoke distribution.

16.2 Regulatory requirements

Airflow visualisation studies are a requirement for the certification of certain types of clean air
devices, as with the ISO 14644 standard, and a regulatory requirement for cleanrooms used for
aseptic processing. With the ISO standard, air-flow studies are referenced in parts 2 (described as
‘airflow visualisation’) and parts 3 (described as ‘flow visualisation’)3. In terms of regulatory

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 CLEANROOM MANAGEMENT IN PHARMACEUTICALS AND HEALTHCARE

requirements, the FDA Guide to Aseptic Processing4 (2004), requires airflow studies for
unidirectional airflow (UDAF) devices, within which product vials are exposed. The Guide requires
that such studies are conducted under dynamic (in-use) conditions. The requirement for airflow
studies is also referenced in the World Health Organisation (WHO) Good Manufacturing Practices
for Pharmaceutical Products5. With the WHO guidance, specific reference is made to the need to
perform airflow studies in Grade A environments in order to assess the uniformity and effectiveness
of the unidirectional flow. In contrast, EU GMP, in Annex 1 for sterile products, does not necessarily
limit the application of airflow studies to unidirectional airflow devices in describing the object of
airflow studies as intended to demonstrate that air does not present a contamination risk to
processing6. The Pharmaceutical Inspection Convention scheme7, to which the EMA and FDA are
members, is least specific of all in simply noting that ‘smoke studies’ need to be performed.

Notwithstanding the subtle differences between the regulatory requirements, there is commonality
in that airflow visualization studies need to be performed within unidirectional airflow devices (ISO
class 5/EU GMP Grade A environments) that are used for aseptic processing and where product
or components are exposed. Within the context of aseptic filling this means, as a minimum, the
aseptic filling zone and where aseptic connections (if any) are performed. Arguably a meaningful
assessment cannot be made without accounting for the impact of the surrounding environment.
Therefore, many airflow visualization studies include an assessment of the cleanroom within which
the unidirectional airflow device is located (ISO class 7 in operation/EU GMP Grade B).

There is a difference in terminology between laminar airflow and unidirectional airflow. It is

important to understand and to define this difference prior to starting an airflow study. The terms
are defined as:
• Laminar flow (also known as streamline flow: An airflow without cross-currents
perpendicular to the direction of flow and no eddies or swirls. The airflow movement is in
single direction and in parallel layers at constant velocity from the beginning to the end of a
straight line vector.8
• Unidirectional flow: An airflow moving in a single direction, in a robust and uniform manner,
and at sufficient speed to reproducibly sweep particles away from the critical processing or
testing area. Being pragmatic, the usage of terminology of unidirectional airflow suits the
industry as it can be achieved at lower velocity than the specified guidance value as well.
In addition to published regulatory standards, important information can be gleaned from a review
of inspectorate trends. FDA 483 notifications concerning airflow studies have indicated 9,10:
• Adequate exhaust systems or other systems to control contaminants are lacking in areas
where air contamination occurs during production. Review of the smoke studies revealed:
• No studies were performed in the sterile filtration room located
• Personnel were not properly gowned
• Only one of the filling technicians was clearly visible
• Pooling was visible between the technician and filling equipment and supplies
• Equipment and supplies normally present in the filling suite were not present during the study
• Air returns were not visible
• Filling activities were not consistent with those observed during a filling operation
• There was no documentation that the studies were reviewed and evaluated
• The technician participating in the smoke study was observed with exposed skin
• The technician's activities were not consistent with those observed during actual sterile
operations
• Pooling and eddying was visible in and around the sterile filling assembly

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 AIRFLOW STUDIES AND AIRFLOW MAPPING

• There was no documentary evidence of in-situ air pattern analysis (e.g. smoke studies)
conducted at critical areas to demonstrate unidirectional airflow and sweeping action over
and away from the product under dynamic conditions. Please note that proper design and
control prevents turbulence and stagnant air in the critical area. It is crucial that airflow
patterns be evaluated for turbulence that can act as a channel for air contamination. The
studies should be well documented with written conclusions, and evaluate impact of
aseptic manipulations (e.g. interventions) and equipment design

16.3 Operational state

Airflow visualisation studies should be conducted with cleanrooms and devices in different states
of operation. These can be divided into the 'at rest' (or static state) and the in operation (or dynamic
state). The static state is with equipment on, but not running, and without personnel present. The
dynamic state is with all machinery normally operating running and with the number of personnel
normally present. Undertaking airflow studies for the at-rest and in-operation states allows the at-
rest state to be compared with the dynamic state study. This can indicate if a problematic airflow in
operation is due to the activity being undertaken or due to the room design. Such comparative
studies also provide important information relating to modifications to the room or HVAC system.
Furthermore, for newly built cleanrooms, the difference between the static state airflow pattern and
the dynamic state are similar if the cleanroom is performing as it was designed11. It is rare, however,
that a static state study alone can be presented to a regulator in lieu of a dynamic state study.

Airflow studies should also be taken at different stages of operation. The activities undertaken
should be representative of standard operations. Such activities maybe simulated where, for
example, the aseptic filling of product is being examined water or other innocuous substance is
normally used due to the risk of product contamination.
After completion of an airflow study, the cleanroom user will need to decide the extent of cleaning.
Although low particulate smoke can be purchased, many types of generated smoke leave a residue
and thus pose a contamination risk. In many circumstances, the room or unidirectional device will
require cleaning and disinfection (with appropriate environmental monitoring) at the cessation of
the activity.

16.4 Smoke generating devices

In order to conduct an airflow visualisation study a smoke-generating device is required. This could
be a smoke pencil for localised, smaller scale examinations, or an industrial smoke generator for
examining a larger area of an aseptic processing room. Such devices are sometimes described as
‘smoke machines’ or ‘fog machines’.

The substance used to generate the smoke is an important consideration. If the substance used to
produce smoke is heavier than air, then there is a risk that the substance could fall out of the air
and may give an incorrect impression that the air is being removed from the room at a faster rate
than is actually taking place in practice. This risk exists more with CO2 based smoke systems than
with glycol-based smoke. Ideally the smoke produced should be of a small particle size as the
smaller the particle size the lower the settling velocity of the smoke (that is the rate at which a
particle will fall, due to gravity). Research has shown, for example that a smoke particle of unit
density, 0.2 micron in diameter, will fall at 8 mm /hr, compared to a 2.0 micron particle, which will
fall at a rate of 468mm/hr. Therefore the smaller the particle of smoke produced, the greater is the
chance that the smoke stays suspended in the air which eventually reduces the occurrence of
deposition of the particles because there is a greater probability of the air being removed from the
cleanroom (through room air changes) than there is of particle deposition.

The most common type of smoke generators uses a water-based fluid which mainly constitutes
pharmaceutical grade glycol and water (either Water for Injections or deionised water). Here

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‘smoke’ is formed as water vapour condenses into tiny liquid water droplets in the air. It is important
that the smoke generated is as clean as possible so that the residual contamination is minimised.
Other types of smoke machine include dry ice machines which create a low lying heavy fog that
uses dry ice (solid carbon dioxide); liquid nitrogen machines (which similarly produce a low lying
fog) and oil-based smoke machines, which leave the greatest amount of residue. Dry ice smoke
machines present some health and safety concerns, especially when used in confined spaces due
to the high volumes of carbon dioxide produced. Smoke pencils normally contain a porous filler
chemical impregnated with fuming sulphuric acid. Therefore when smoke particles are emitted from
the end of the tube they have acidic properties due to sulphuric acid aerosols. The smoke produced
must conform to national health and safety requirements.

Table 1 displays the most commonly available types of devices for generating smoke for airflow
studies together with the advantages and disadvantages of the methods.

Table 1: Smoke generating devices for conducting cleanroom airflow patterns

Type of smoke generator Advantages Disadvantages

Smoke pencil Inexpensive Low volume of smoke
Easy to use Acidic smoke (leaves residues
that can damage some materials)
Glycol/water High volume of persistent Leaves residues (water soluble)
smoke
No safety concerns
Dry ice High volume of smoke The smoke is heavy and can
Liquid Nitrogen No residues alter the test results
Some safety concerns

As an alternative to hand-held smoke generation devices, smoke generators can be ducted. This
allows for a steady flow of smoke to be produced (in a curtain like effect) (Figure 1). This can be
useful for examining larger aseptic filling machines where the object is to note any ingress from the
external air into the critical zone (sometimes referred to as the ‘work zone’). To achieve the required
distribution of smoke this can function either through the inherent velocity of smoke produced by
the generator or by incorporating fan assistance where a rapid distribution of the smoke is required.

For ducting, this should be as short and straight as possible with as smooth a bore as practical, so
that frictional losses within the duct are kept to a minimum. Ideally the smoke output should be
controlled so that a low output can be
used to show unidirectional flow and a
higher output used where the impact of
the surrounding turbulent air upon the
unidirectional flow is required.

Glycol/water smoke generation devices

work by heating a chemical (such as
propylene glycol or triethylene glycol,
mixed with 20 percent water) above its
boiling point within a heat exchanger. The

Figure 1: Example of ducted smoke

through a perforated pipe in order to
obtain a curtain-like effect.

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 AIRFLOW STUDIES AND AIRFLOW MAPPING

chemical is then vaporised and as the vapour passes out of the heat exchanger and starts to mix
with the relatively colder atmospheric air, the rapid condensation of the vapour results in a visible
smoke (or technically the more accurate term ‘fog’). The configuration of the heat exchanger is
important in order to avoid smoke of too large a particulate size being produced and thus avoiding
the production of smoke of variable quality. If the heat exchanger becomes too hot the machine will
malfunction; if the heat exchanger becomes too cold it will produce liquid rather than smoke.

Smoke pencils are small, chemical-filled glass tubes. When both tips are snapped off and air is
squeezed through the tube, a jet of white chemical 'smoke' is produced. Smoke pencils are best used
for small, localised studies, such as for detecting air leaks in ductwork, cracks around doors, etc.

16.5 Video recording

Airflow studies should be captured by video recording. This not only captures all the required
information but also provides a better historical record than can be provided from photographs or
from hand drawings. The other advantage of video recordings is that they can be shown to auditors
and enable the findings of the airflow study to be communicated explicitly and easier than is
possible through diagrams or written descriptions. The use of video tapes or DVD recordings is
generally a regulatory expectation.

Care must be taken when using video recording that a suitable video camera is used, with sufficient
definition to allow for the smoke to be seen clearly on the finished recording. The use of a tripod is
recommended so that any shaking of the camera by the camera operator is minimised. It is also
important that the recording is secure and stored in a way which does not damage the digitally
captured images for future playback. Making a second copy or transferring onto a computer hard
drive or server is recommended.

16.6 Undertaking an airflow visualisation study

As stated earlier, before starting an airflow visualisation study the area to be examined must be
prepared as normally in terms of equipment, personnel and simulated activities. The staff present
should wear appropriate cleanroom clothing as if the event were an actual aseptic operation, even
if the cleanroom is temporarily declassified, otherwise the completed recording will not indicate to
a regulator the full extent of normal operations. Often regulators use the airflow visualisation
studies also for the evaluation of the aseptic technique, since many activities performed in the
critical area (example: some equipment set-up operations) are not easily visible from the external
windows, therefore a great attention to this aspect must be posed during the video takes.

It is generally a good idea to have two people, separately from what is happening in relation to the
process, to run the airflow study (although care must be taken to avoid these additional people from
disrupting the study itself). One person is tasked with operating the smoke generator and for
‘directing’ the study, and the second person is tasked with operating the video camera.

It is important that, before the study starts, that a script is written and consideration be given as to
what will or will not be an acceptable outcome (this is largely a qualitative assessment). This will
ensure that the people who will be recording the study and generating the smoke know what they
should be looking for. This will include consideration of ‘what are the activities being performed? ,
‘how many times with any activity take place?’, ‘how long will the study last for?’ and so forth. The
person who will be operating the video camera will need to understand when close-up shots are
required (such as, examining the impact of the airflow around a filling needle or for an intervention
involving the use of tools) and when a wider-angle shot is needed (to show the path that the smoke
takes). Furthermore, it may be necessary to shoot the same activity using different angles so that
all aspects of the activity and smoke distribution can be seen. Care should be taken when filming
to avoid glare and reflection from the cleanroom lights reflecting from certain surfaces or glass, and

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situations when personnel move or stand immediately in front of the camera. The use of a polarized
filter on the camera lens can help in reducing reflections and glare. In some situations the smoke
may not be clear against a white background and it may be necessary to have a darker material
(such as a dark plastic material fixed to the wall) in order to provide sufficient contrast12. In some
cases, additional light sources may be useful in increasing the overall contrast, mainly if the light
beam is orthogonal to the direction of the video capture (the Tyndall effect).

When conducting the study, the person who operates the smoke generating device must be careful
not to introduce large volumes of smoke into an air stream, otherwise the smoke will either be too
heavy and it will be unclear as to what is happening with the airflow, or that the velocity of the
smoke will be too great and the airflow disrupted. Instead the smoke should be introduced gently
at a low velocity so that the smoke produced is steady and ‘light’, to the extent that the smoke
should appear to float in the air and then dissipate leaving little residue. It is equally important that
the smoke remains visible for the required time.

The use of a video monitor or a PC directly connected to the digital video camera during the video
takes can help in evaluating immediately if the recording is meeting the expectations. This is not
easy on the small screen of the video camera. Considering the efforts implied in an airflow
visualisation study, it could be disappointing to discover that a video take is not satisfactory at a
later stage, when repeating the recoding could be problematic.

16.7 Key objective of airflow visualisation studies

There are a number of essential aspects of the airflow that need to be examined by conducting an
airflow study. When examining aseptic processing, the primary aim is to indicate if there is
adequate protection of exposed product, product contact components and surfaces. Other
considerations include:
a) To understand the difference between static and dynamic states and the direction of air
laminarity away from critical zone. Here it is important to establish if the airflow is smooth, free
from disturbances (such as small, temporary vortices or eddies) and unimpeded. The direction
of the air pattern should be noted. The question to frame analysis is: Is the air movement as
expected? For example:
i) Is the air avoiding the floor?
ii) Is the air moving into a lower grade area?
At rest, airflow studies can also be used to make the following important observations:
a) Demonstrating that airflow patterns do not present a contamination risk, such as considering
if the air from background environment is getting mixed with the air stream from the UDAF
source. It should be proven if the clean zone is positively pressurized in relation to the
background environment. During this test the “induction” effect can also be evaluated.
b) Evaluation of routine operational conditions can be examined, such as changes to the
airflow during one or several door opening of the background clean room.
c) To evaluate the unidirectional airflow from the airflow device. The Institute of Environmental
Sciences and Technology (IEST) and National Environmental Balancing Bureau
recommends maintaining not more than a 14° angle of deflection from vertical.
d) To evaluate the installation and operation of equipment beneath the airflow and to evaluate
impact of equipment on the unidirectional airflow.
e) To evaluate the ‘sweep away’ action of air during the operation of the filling/ stoppering and
sealing stations, and its effect on the product containers or closures.
f) Evaluation of airflow between the stoppering and capping station should be performed in
order to ensure that air flow is from stoppering station to capping (lesser clean/particle
generating) station is satisfactory.

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g) To evaluate the interfaces with the filling line. For example a depyrogenating tunnel (the
exhaust fan of the depyrogenating tunnel shall be designed to exhaust air from the filling
zone isolator as well, since airflow should be from filling station to cool zone of tunnel),
freeze-dryer, mousehole (the exit path for vials from isolators is termed as a mousehole.
The isolator may have other exit ports through which product is discharged, opening the
isolator to the outside environment. Airflow at this station should be towards the outside,
which should also be justified with sufficient overpressure in a continuous manner to
ensure that isolation is maintained at this location), etc.
Prior to the actual test for airflow pattern, a trial run may be conducted by keeping to the above
objectives with clearly defined acceptance criteria. This will help in optimizing the operational
parameters of the equipment and surrounding environment. Operating parameters such as speed
of either inlet or exhaust fans (Proportional Integral Derivative (PID) controlled fans), pressure
differential between two stations in the clean zone, room pressure differential, etc.
b) Within cleanrooms, where a turbulent flow is expected (an air-pattern characterised by small and
temporary fluctuations caused by instabilities) the impact of any air eddying must be examined
to note any impact upon critical process steps. Within cleanrooms, the air streams should be
quickly dispersed. Areas of poor air movement should be noted. A combination of single 'puffs',
'streams' or 'multiple streams' of smoke can be used to investigate possible problems.
With both ‘a’ and ‘b’ above, examples of problems or points of concern that may be found include:
• Heat rising from machinery and disrupting the airflow
• Obstructions preventing the supply of air getting to the critical area
• Obstructions, or machinery design, turning a unidirectional flow into a turbulent flow
• Entrainment: contaminant drawn into the clean air
• Stagnant or turbulent areas acting as conduits for entry of contamination from areas of
lower contamination
• Air flowing from personnel towards the product
• Examination of HEPA filters to detect any adverse affect on air flow patterns, which will
eventually have an affect on unidirectional airflow
• Evaluation of disruption of Active Air Sampling/particle counting in the clean zone
c) To demonstrate the impact of operator interventions and other personnel activity, including
environmental monitoring (the placement and operation of active air-samplers can be
particularly disruptive to the air pattern). Studies of interventions must take into account:
1) Prime objective of such studies is to demonstrate that air-flow patterns do not present a
contamination risk during performing routine interventions e.g. care should be taken to
ensure that air flows do not distribute particles from a particle generating person, operation
or machine to a zone of higher product risk. Insertion of hand into all the gloves of a
positive pressure isolator should also be considered.
2) Such studies must ensure that that, during performing of any intervention, there is no
obstruction to the airflow over the container/closure/product is in open condition. All the
interventions shall be evaluated against this criterion by performing and assessing the air
flow pattern.
3) It is important that UDAF devices are operating as normal while performing interventions in
the clean zone.
Airflow pattern studies that examine interventions can be conducted prior to media fill simulation
runs to finalize the modus operandi of aseptic interventions. These studies are helpful in
developing the interventions in a complaint manner. Video records of these studies can also be
made in order to train the technicians involved in machine assembling, filling, and stoppering
activities. The impact of other personnel operations should be assessed as appropriate;

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d) To show the impact of equipment operating. One important observation which can be made is
assessing the impact of a change in vial size (minimum or maximum can be considered) in
order evaluate the air flow along different stations in a filling line. This can be assessed at
different states: at the start of batch (front line is empty and impact of automatic opening of
tunnel door); during batch (vials completely filled in the line) and at end of the batch (rear side
is getting empty).
e) To demonstrate the set-up of equipment
f) The area outside of the UDAF units (within the ISO Class 7/EU GMP Grade B cleanroom).
Arguably, the biggest contamination risk in these areas is when potential airborne particles
could accumulate in vortex regions. This can happen when an unidirectional air-flow strikes an
object and creates a ‘wake region’. It is prudent to confirm the level of particles in such
regions through the use of a discrete particle counter.

In general, when examining airflows the streamlines should be noted. A streamline is defined as
the route the air is taking and a streamline will be the path that any contamination by ventilation
could take (so-called convective transport). When a streamline meets an object and causes
turbulence (or meets air which is turbulent), then contamination could potentially be dispersed (or
so-called turbulent diffusion). Regions of stagnation should also be noted. These can often occur
in front of machinery and by work surfaces that are perpendicular from the airflow. The air velocity
in these areas can be unpredictable.

16.8 What to include in airflow studies

There is a range of aspects of an airflow that should be examined at the time the airflow is conducted
and when playing back the video recording13. These items are listed and discussed below:
a) Examination of the unidirectional airflow.
The unidirectional airflow is a key parameter of a clean air device in terms of the air velocity
(operating at 0.45 metres per second (±20%) and direction14. These devices contain HEPA
filters, which control the air speed and direction. The object of the unidirectional airflow is to
push outward any contamination which might be deposited into the air-stream and to avoid
the potential for contamination dropping out of the air, either though gravity or by striking a
object, and falling onto a critical surface. To measure the aerodynamic performance of the
unidirectional airflow unit, smoke should be introduced at the filter face so that the
distribution of the smoke downwards and away from the critical zone can be seen. Smoke
should also be introduced at the working height, immediately above the area where product
or product components are exposed.
The assessment should note the impact of the machine upon the airflow. Does the smoke, for
example, entrail inwards when the air impacts upon the filling machine guarding? What is the
effect of disturbances caused by the motion of machine operations? The biggest risk will be
when potential airborne particles accumulate in vortex regions. When a unidirectional air flow
strikes and object, an obstacle will create a 'wake region'. Such regions should be studied for
vortices and potential particle accumulation.
Any regions of stagnation should be detected. When unidirectional airflow meets an object,
wakes and vortex streets can be formed. This causes turbulence that can lead to pockets of
stagnation in front of machinery and work surfaces that are perpendicular to the main
direction of the airflow. Such pockets can be unpredictable in speed and direction and
require mapping. Another potential risk from large surfaces is that wake regions can entrain
ambient air into clean zones. A consideration of the impact of Grade B areas upon Grade A
zones should be considered in such circumstances.
Ideally, the pattern should show that the air is characterised by a smooth flow, free of any
disturbances (such as small and temporary vortices or eddies) and unimpeded.

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b) Examination of the operation of the aseptic process.

Where a filling machine is examined, for example, the machine should be filmed running with
bottles or vials moving along a conveyor belt. The filling operation should be simulated with
filling needles moving upwards and downwards. Ideally a placebo should be used in place of
product, such as filling vials with water.
For filling machines, steps such as machine set up, loading of vials, stoppers and caps;
cleaning of spilled product; cleaning vials; removing jammed stoppers; gauntlet changes, etc.
should be captured in the study.
c) Examination of in-process interventions.
Where interventions by personnel are commonly performed during aseptic processing, these
should be simulated during the study. This may include activities like picking up fallen vials
using forceps, adjusting stopper grippers, fixing filling needles, etc. The air patterns should be
carefully assessed for turbulent regions where contamination may be dispersed in an
unwanted direction or accumulate15. In ideal circumstances, the airflow should continue to flow
out of ISO Class 5/Grade A and into ISO Class 7/Grade B. Where excessive turbulence is
created and air flows back from ISO Class 7/Grade B into ISO Class 5/Grade A, this will
indicate a potential problem as shown in Figures 2 and 3.

Grade B Grade A Grade B Grade A

Filling Room Filling Machine UDAF Filling Room Filling Machine UDAF

Figure 2: Representation of an intervention by an Figure 3: Second representation of an

operator into an ISO Class 5/Grade A zone. intervention by an operator. Individual performs
Individual performs intervention. Very little intervention. Turbulent airflows are formed
turbulence is generated and air stream is from resulting in some air being driven back into
Grade A to Grade B. This situation is satisfactory. Grade A. This situation is not satisfactory.

In addition the effect of the

operator breeching the barrier
between the critical zone and
the cleanroom (such as
between ISO class 5 and class
7) should be noted.

Figure 4: Typical smoke pattern

when a Grade A filling machine
cabinet window is opened (note
the additional light behind the
operator to enhance the smoke
visualization).

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d) Examination of the cleanroom.

The room in which the unidirectional airflow device is located must also be studied. This is
important in order to determine if there is any impact of the room upon the unidirectional device
(particularly any possibility of air ingress either in standard operation or where the filling
machine doors are opened). It is also important to study the effect of personnel and equipment
movement within the turbulent flow16, in terms of general contamination control (this may
indicate, for example, where there is dead air, particularly arising from local impediments 17 and
thus any areas of potential contamination). It is important to see if the air in a cleanroom is
supplied and mixed well to ensure that all contaminants are removed. For room studies, smoke
should be delivered at the HEPA filter face and its distribution across the room noted.
Ideally, the air pattern will show that the airflow is characterised by small and temporary
fluctuations caused by instabilities. The flow is not necessarily constant and will fluctuate
around an average value. Then will be small and temporary eddies. Such patterns will occur
under UDAF when air meets an object. The impact of the temporary eddy should be
assessed. Some surface materials can attract airborne particles base on the quality of the
material (such as roughness, electrostatic properties). Where this occurs, the impact and
potential concentration of contamination should be considered as appropriate. The
contamination rate will depend on:
• Concentration of contaminants and motion of contaminants
When cleanrooms are examined, the effect of the ‘clean-up’ time can also be measured.
Here the room can be filled with smoke and then the time taken for the smoke to clear (as
it is extracted and the room replaced with clean air from the HVAC system) captured. This
part of the study will also indicate if there are any areas of the cleanroom that clear more
slowly than others. Such information can be useful for future reference should a problem
with particle counts occur in the future as the time taken for the room to clear is known.
d) Dynamic characteristics of the door opening and closing operation18.
Doors of filling rooms and filling machines should be opened in order to simulate what might
happen during processing. With filling rooms, if equipment is required to be transferred during
an operation, this activity should be simulated. When examining the impact of door opening,
the extent of backflow velocity, and thus the potential for less clean air ingression into a
cleanroom, should be noted. This can vary depending upon the type of cleanroom door.
Information relating to the air distribution in the cleanroom can also provide supporting
information about the function of pressure differentials. Most importantly, there should be a
cascading positive air pressure differential system, where the “cleanest” space is maintained
at the highest pressure and the lesser cleaner spaces decrease in pressure. For this the
impact of the pressure gradient upon the airflow can be examined. The function of increased
pressure is to force clean air outwards, thus preventing the ingress of contaminated outside
air entering the clean zone. A pressure differential also prevents the entry of outside air when
the door is open.

16.9 Reviewing airflow studies

Once complete, airflow studies should be reviewed against the pre-set acceptance criteria for the
clean air zone, room or activity. The airflow pattern study report should contain details about the
operating parameters and set points for the operating equipment involved in the study. The
set/operating parameters could be the inlet fan speed, the exhaust fan speed, position of automated
air control valves, pressure differentials, pressure differential between tunnel and isolators, etc.
Problems should be noted and preventative actions considered. The most effective actions are
those that lead to a change in practice or redesign, so as to eliminate the risk. Only where risks
cannot be eliminated should monitoring be targeted19. Where significant changes take place, the
airflow study should be repeated so that the effectiveness of the action can be measured.

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 AIRFLOW STUDIES AND AIRFLOW MAPPING

Examples of problems often detected during airflow studies are reported in the table below with
possible corrective actions:

Table 2: Common concerns from airflow visualisation studies and suggestions for
corrective actions
Problem
Low air velocity inside a Grade A/ISO class 5
cabinet at the working height (vertical flow).
Stagnant smoke pockets at the bottom side
of the cabinet.
The air pattern inside the Grade A/ISO
Class 5 is not unidirectional. There are
turbulences immediately downstream the
surface of the HEPA filters.
There are turbulences at the interface
between the Grade A/ISO class 5 cabinet
(filling machine) and the adjacent pieces of
equipment (example: depyrogenating tunnel,
crimping machine)
Possible corrective action
This problem is often caused by insufficient air
exhaust at the bottom level of the cabinet windows.
The windows should have openings at the bottom
side big enough for allowing an easy exhaust of the
air coming from the HEPA filters.
This can be caused by obstacles hidden behind the
HEPA filters grid (lights, grid frame). Remove the
obstacle; use lights specifically designed for Grade
A/ISO class 5 cabinets.
Another root cause for this kind of problems could
be a not uniform velocity of the air coming from
every single HEPA filter units installed in the Grade
A/ISO class 5 cabinet. It is necessary to check the
air velocity of each filter with a calibrated
anemometer and replace any partially clogged filter.
Turbulences in these areas are almost unavoidable.
It is important that the airflow study confirms that
the air is always flowing from the inside of the most
critical area (the filling machine) towards the
adjacent areas. If the turbulence is too evident it
can be minimized by differential pressure
adjustments.

16.10 Documenting airflow studies

As for other qualification/validation activities, it is advisable that each airflow visualization study is
executed following a pre-approved protocol. The protocol is also useful for planning accurately all
the activities necessary for an effective execution of the study. Executing an airflow study is a
relatively complex and time consuming activity that can require several days of work and
consequent shutdown of the involved production area. Therefore it is very important that all the
activities are appropriately planned and that all the required materials are available when needed.
For example, it would be wise to have available a back-up for critical instruments (smoke generator,
video camera) and plenty of consumables (dry ice, liquid nitrogen, or glycol solution), since it is not
easily predictable how many times a specific production step could need to be repeated (mainly if
it the first study for a specific department/line).

An example of content of an air flow study protocol could be:

1. Title and approval page
2. Scope
3. Description of the area/equipment to be tested and selected conditions (use of a risk
assessment could be advisable for selecting the appropriate conditions, such as: vial size,
line speed, number of operators present, etc.)
4. Responsibilities
5. Instruments/materials
6. Activities to be covered (for example):

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 CLEANROOM MANAGEMENT IN PHARMACEUTICALS AND HEALTHCARE

a. Filling line set-up

b. Filling line in static conditions after set-up
c. Filling line in dynamic conditions
d. Simulated interventions
e. Environmental Monitoring
7. Acceptance criteria
8. Cleaning of the line and environment post execution

For each activity listed at point 6., a more detailed description of each single step to be covered
should be included in the protocol, the execution of the activity must be similar to what happens
during the aseptic filling simulation studies (Media Fill).

An example of a form to be used for describing the activities to be executed for simulating critical
interventions and then for documenting their execution is attached (see Appendix I).

16.11 Reporting airflow studies

Once completed, the airflow visualisation study should be copied to permanent media (such as disc
or computer hard-drive). The study should be given a unique reference number (or the protocol
number) and the date and time of the study should accompany the recording. It is not advisable to
edit the master study (original recording), as this could create suspicion to a regulator that the study
has been adulterated. If a scene has been shot incorrectly, this should be noted in the final report.
However, editing the study (example on a DVD divided in chapters) can be helpful for presenting
it to the inspectors provided that the “master recording” remains unaltered in the event that a
regulatory inspector wishes to view it. Furthermore, narration can be added to the recording if it is
considered helpful, although in most circumstances this will be unnecessary if the accompanying
report is written sufficiently well. Adding music to the recording is an unnecessary frivolity and
should be avoided.

The study should then be reviewed by a competent person and a report generated. The amount of
detail which goes into the report will depend upon the user (to the extent that the report describes
each scene). Nonetheless, notable events should be described with reference to the time or frame
number of the recording.

In compiling the report, the results of airflow studies should be carefully examined to see if the
design of the room is appropriate and to determine if the activities conducted in the area are
suitable for minimising airflow disruption (and where disruption occurs this will lead to decisions
being required whether the design needs to be changed or the activities modified).

In addition, airflow studies should be considered in relation to the environmental monitoring

undertaken within the cleanroom or unidirectional devices under normal operations. Where there
is dead air or risk of turbulence which cannot be easily eliminated, then it is important that
environmental monitoring takes place. This will involve consideration of the use of settle plates or
active air-samplers (noting the concern that active air-samplers can themselves disrupt the airflow).
The selection of sites for environmental monitoring should be documented 20.

16.12 Frequency of airflow studies

Airflow studies are normally undertaken for newly built cleanrooms and after significant
modifications. The airflow studies presented to regulatory inspector should reflect normal
operations.

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 AIRFLOW STUDIES AND AIRFLOW MAPPING

One consideration is the frequency of repeating airflow studies so as to demonstrate a continuous

state of control. The user will need to decide if airflow studies are undertaken only after significant
modifications have taken place or whether they are carried out at regular intervals (as with the
requalification of other critical cleanroom parameters). The frequency of requalification would need
to be determined by some type of risk assessment.

16.13 Alternatives to airflow studies

Computational design systems are sometimes used as alternatives to airflow studies. Such
approaches are more common within the microelectronics and semiconductor industries than with
pharmaceutical cleanrooms (nevertheless such approaches can be used during the design phase
for a new cleanroom). Embarking on alternative and novel approaches is more difficult in
pharmaceutical cleanrooms because of the pharmaceutical stringent regulatory requirements for
an airflow study.

Alternative approaches include numerical flow and particle transport modelling21. Here
comparisons can be made between experimental data from an operating clean room and
theoretical predictions indicate that the model is sufficiently accurate to predict the main features
of the flow and particle transport for various configurations and operating conditions 22.

16.14 Conclusion
This chapter has examined the importance of airflow studies as a means to visually understand the
airflow within a cleanroom or clean air device, and has indicated that understanding the air pattern
is important in order to know the level of contamination risk and to identify any potential weak areas
within cleanroom design and operations. This chapter has also provided pragmatic tips in
conducting a meaningful airflow study that will satisfy the regulatory considerations.

Although airflow visualisation studies are a necessary part of cleanroom assessments and can
reveal valuable information they do not provide the complete picture of contamination concern or
allow a complete assessment of contamination risks to be made. As Ljungqvist and Reinmüller
have pointed out, for example, airflow studies must be combined with other observations and
environmental monitoring data in order to entirely understand the cleanroom risks23.

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 CLEANROOM MANAGEMENT IN PHARMACEUTICALS AND HEALTHCARE

Appendix I
Example of a form for documenting an airflow study covering critical interventions on a filling line
to be included in the study protocol. Similar forms may be used for the other steps (e.g. equipment
set-up, environmental monitoring, etc.).

It is based on the use of a Digital Video camera (DV tape recording). The indication of the time
code (that identifies every single frame) allows for easy retrieval of each simulation step. It is
important that the date and time of the camera internal clock are synchronised with the local time
in order to document on the video recording the exact time of the video takes.

Filling Line ________________

(Critical interventions to be executed)
Operation to be executed/simulated Video recording data
Tape # Time code Time code Done by
start end
Vial removal from the accumulation table
at the exit of the tunnel using the sterile forceps.
Stopper removal from the guide using the
sterile tool.
Dosing pump and needle replacement
(opening window A).
Vial removal from the filling line with sterile
forceps (opening window B).
Simulation of line cleaning and sanitisation
after vial breakage.

COMMENTS
Executed by _______________________________ Date___________________
Reviewed by _______________________________ Date___________________

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16.15 References
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Jinno N. Study of airflow pattern in the clean room for manufacturing semi-conductors. Proceedings
of the Institute of Environmental Sciences 1986:556-564.
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Sandle T. Environmental Monitoring. In: Saghee MR, Sandle T and Tidswell EC. eds. Microbiology
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Horizons, 2011, pp293-326.
3
International Organisation of Standardisation. Cleanrooms and Associated Controlled Environments.
I. Classification of air cleanliness; II. Specifications for testing and monitoring to probe continued
compliance with ISO 14644-1; III. Metrology and test methods. BS EN ISO 14644 Parts 1,2 and 3.
1999 (Part 3: revised 2005).
4
Food and Drug Administration. Guideline on Sterile Drug Products Produced by Aseptic Processing
FDA, Rockville, MD, USA, 2004.
5
World Health Organisation. WHO GMP for Sterile Pharmaceutical Products, Working document
QAS/09.295 Rev.1, Geneva: World Health Organisation, 2009.
6
European Commission. EU GMP Guide to Good Manufacturing Practice. Revised Annex 1:
Manufacture of sterile medicinal products. Brussels, Belgium. European Commission, 2008.
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Pharmaceutical Inspection Convention (PIC/S). GMP Annex 1 Revision 2008 Interpretation of the
Most Important Changes for the Manufacture of Sterile Medicinal Products – Recommendation,
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Batchelor GK. Introduction to Fluid Mechanics. Cambridge: Cambridge University Press, 2000.
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GMP Trends Inc. GMP Trends Issue 20714. GMP Trends, Colorado, USA.
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GMP Trends Inc. GMP Trends Issue 20716. GMP Trends, Colorado, USA.
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Kuehn T, Marple VA, Han H, Liu D, Shanmugavelu I,Yourself SW. Comparison of measured and
predicted airflow patterns in a clean room. Proceedings of the Institute of the Environmental
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Klinberg S. Smoke studies: Clearing the mystery of air flow visualisation. Journal of Validation
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13
Ross S and Sandle T. Air pattern analysis of a filtration transfer. Pharmaceutical Microbiology
Interest Group News 2007,26:3-4.
14
Gail L. Facts and fiction in cleanroom metrology. Journal of IEST 2010;53(1).
15
Ljungqvist B and Reinmüller B. Risk assessment with the LR method. Eur J Parenteral Sci 2002;
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16
Boyle DR and Golay MW. Measurement of a recirculating, two-dimensional, turbulent flow and
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17
Settles GS, Huitema BC, McIntyre SS and Via GG. Visualization of clean room flows for
contamination control in microelectronics manufacturing. In: Flow visualization IV: Proceedings of the
Fourth International Symposium, Paris, France, Aug. 26-29, 1986 (A87-52301 23-34). Washington,
DC, Hemisphere Publishing Corp., 1987, pp 833-838.
18
Shigeo H, Kazutomo I, Yoshiaki K, Kaoru M. Dynamic characteristics of the door opening and closing
operation and transfer of airborne particles in a cleanroom at solid tablet manufacturing factories.
Transactions of the Society of Heating, Air-Conditioning and Sanitary Engineers of Japan
2004;95:63-71.
19
Katayama H, Higo T, Tokunaga Y, Hiyama Y, Morikawa K. Establishment of critical contamination risk
locations ("hot spots") in environmental monitoring by means of three-dimensional airflow analysis
and particulate evaluation. PDA J Pharm Sci Technol 2005; 59(1):49-63.
20
Sandle T. Environmental Monitoring Risk Assessment. Journal of GXP Compliance 2006; 10(2):54-73.
21
Murakami S, Kato S and Suyama Y. Three-dimensional numerical simulation of turbulent airflow in a
ventilated room by means of a two-equation model. ASHRAE Trans 1987; 93(2):621-641.
22
Kuehna T H. Clean room aerosols: Predicting air flow patterns and particle contamination in clean
rooms. Journal of Aerosol Science,1988; 19(7):1405-1408.
23
Ljungqvist B, Reinmüller B. The LR method in critical areas: airflow patterns and the design of
aseptic interventions.

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