24-03-2019

FAQs ON PARTICLE COUNTING

1 Minimum Sampling Flow Rates &

Volumes?

2 Maximum Permissible Sampling Tube

Length

3 How to conduct Recovery Test?

by Sheesh Gulati
Secretary, ICCCS
Chairman, CCSI

FAQs ON PARTICLE COUNTING

What is the Minimum /

Maximum / Recommended
Flow Rate specified in
Cleanroom Standards?

FLOW RATES
• Airborne particle counters are available with
flow rates of 0.1cfm,1.0 cfm, 50 lpm or 100
lpm.

• Most standards do not specify using a

particular flow rate. The new ISO14644
standard only specifies a flow rate of 1 cfm for
counting particles >=5 microns. So what flow
rate should you use?

• The following would be the problems using a

low flow rate 0.1 cfm counter while
validating pharmaceutical clean rooms:

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Flow rates (contd.)

– We would have to multiply the 1 minute
measurement result by 10 to get counts per
cft. And again by 35.2 to get counts/cu metre.
This is a valid approach provided that the
measured value has statistical significance i.e. it is
20 counts per minute or greater to provide a
reliable basis.
– If the measured value was only 3 particles in 1
minute measured with a 0.1 cfm counter, this
would convert into 30 particles/cu.ft. but if count
changed to 4 particles it would jump to 40/cu ft.

FLOW RATES (Contd.)

– The other method would be to let the 0.1cfm counter
run for 10 minutes to get a sample volume of 1 cubic
foot. The draw back of this approach is that, if any short
term particle burst occurs during the relatively long 10
minutes count cycle, this burst would be integrated and
would not show up as significant.
– The best way to count large particles is to sample a
large quantity of air, thus maximising the chances of
capturing any 5 micron particles that may be present.
Therefore 1 cfm flow rate is better than 0.1 cfm as
you get 10 times the sample quantity

Effect of Flow Rate on

Particle Sizing

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Ultra High Flow Considerations (100 lpm)

• Impact on immediate environment and airflow
patterns
– Isokinetic probe must be matched to particle counter
flow rate
– High exhaust rate of sampled air may disturb room air
flow

• Disturbance of airflow patterns in restricted areas

due to high rate of sampled air
– Modest air supply of most LAF cabinets may not be
sufficient to support high flow rates without substantial
disturbance
Pulsations in high flow pumps cause inconsistent air flow within
the counter resulting in poor size resolution performance
Long term performance – higher maintenance requirements

Smoke Studies
(Impact of exhausted air)
Particle Counter Off
Photo shows horizontal
laminar flow across table

Particle Counter On
Photo shows disruption of
laminar flow near the
particle counter exhaust
The greater the flow rate of sampled
air, the greater the amount of
Exhaust Air

DON’T SACRIFICE SENSITIVITY FOR SPEED !!

• If you study the specifications of most particle counters
with 100 lpm flow rate, one fact clearly stands out. The
smallest particle size they can measure is only 0.5
microns, not 0.3 microns.
• And the counting efficiency for the most sensitive
threshold of 0.5 microns is 50%. In other words,
compared to a theoretically perfect counter, they are
only counting half the particles of 0.5 micron size!!
• Counting efficiency is an expression of the probability
that a Particle Counter will sense and count a particle
passing through its sample volume. This probability is a
function of size up to a certain critical size above which
all particles are normally sensed and counted.

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FAQs ON PARTICLE
COUNTING
What is the Minimum
Sample Volume to be
collected?

Sample Volumes per ISO 14644-1:2015

No change in minimum sampling volume and time in 2015 version

Clause A.4.4: “At each sample location, sample a sufficient volume of air that a
minimum of 20 particles would be detected if the particle concentration for the
largest considered particle were at the class limit for the designated ISO Class”

Vs = 20_ X 1000
Cn,m
Vs is the minimum Sample Volume Per Location (in Litres)

Cn,m is the class limit (particles / M3 ) for the largest considered particle size specified for
the relevant class

20 is the defined number of particles that could be counted if the particle concentration
were at the class limit

“The Volume Sampled at Each Location Must be at Least 2 Litres,

With a Minimum Sample Time at Each Location of 1 Minute”

What volume do I need to

collect?
ISO 14644 requires sample with 20 theoretical particles
ISO 14644 allows 3520 particles (0.5um) in 1m 3 (1000 litres)
for Class 5

20 particles
Volume = X 1000 litres
3520 particles

Volume = 5.7 litres

At sampling rate of 1cfm (28.3 lpm) this would take less than one minute

Greater than 2.0 litres so minimum sample volume is satisfied

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Minimum Sample Volume

Regulations and Guidance for the Manufacture
of Sterile Medicinal Products ISO 14644 and
EU GMP Annex 1
EU GMP Annex 1:2009 states:
“For classification purposes in Grade A
zones, a minimum sample volume of 1m3
should be taken per sample location”

This is sometimes mistaken to imply that

1 cu metre sample per location is
required even during routine monitoring
of the Grade A zone

EU GMP Annex 1 2009

EU GMP Clearly differentiates classification and monitoring
Section 4.1 Clean room / clean air device
classification
General interpretation: The GMP Annex 1 Revision
distinguishes very clearly between clean room / clean air
device classification which is described in sections 4 to 7,
and clean room monitoring, which is described in sections
8 to 20.
In general, clean room/clean air device
classification is required to be performed according
to ISO 14644-1 with the applicable limits for
particle counts defined in the table in section 4 of
GMP Annex 1. Monitoring, on the other hand, does
not need to be performed according to ISO 14644-
1. It can be performed for a reduced number of
sampling points and sampling volumes.

EU Annex 1 : Monitoring
Section 12:
“The sample sizes taken for monitoring purposes using
automated systems will usually be a function of the
sampling rate of the system used. It is not necessary for
the sample volume to be the same as that used for formal
classification of clean rooms and clean air devices.”

• It is not necessary to sample 1m3 during verification

or monitoring
• Particle counters used for monitoring may have the
same or different flow rate from those used for
classification.

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FAQs ON PARTICLE
COUNTING
What is the Maximum
Permissible Sampling
Tube Length?

SAMPLE TUBING LENGTH

• Particle loss in the sample tubing applies more to larger
particles of 5 microns and larger, and less to smaller
particles of 0.3 and 0.5 microns.The inertia of larger
particles makes them bump into the wall of the tubing
and they may stick to the tubing wall.
To lessen this problem, two methods are recommended:

• Flow rate should be fairly high i.e. 1 cfm not 0.1 cfm.
The higher the flow rate, the lesser the number of larger
particles that will be "lost"
• Sample tubing length should be minimised, although
tubing cannot be totally eliminated.
• In the case of portable particle counters, the tubing
length should not be more than 3 metres i.e. 10 ft

Sample Tubing Length

EU GMP Annex 1 states:
"Portable particle counters with a short length of sample
tubing should be used for classification purposes
because of the relatively higher rate of precipitation of
particles ≥5.0µm in remote sampling systems with long
lengths of tubing."

and

"Where remote sampling systems are used, the length of

tubing and the radii of any bends in the tubing must be
considered in the context of particle losses in the tubing."

You will notice that nowhere is the "short" length specified.

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Sample Tubing Length

ISO 14644-3 Annex B Test methods Clause
B1.2.2 states:
"The transit tube from the sample probe
inlet to the Discrete Particle Counter
sensor should be as short as possible. For
sampling of particles larger than or equal
to 1 µm, the transit tube length should not
exceed the manufacturer's recommended
length and diameter“
Most particle counter manufacturers
recommend a maximum tubing length of 3
metres

Particle Sampling Tubing

Fed Std 209E: Obsolete Since 2001: But Helpful:

►B40.2.1 Particle transit considerations. The probe and

transit tube should be configured so that the Reynolds
number is between 5,000 and 25,000.

►For Particles in the range of 0.1 to 1.0 µm and a flow rate

of 0.028 M3/min (1.0 ft3/min), a transit tube up to 30 m long
may be used.

►For particles in the range of 2 to 10 µm the transit tube

should be no longer than 3M.

Particle Sampling Tubing

ASTM F50: Standard Practice for Continuous Sizing
and Counting of Airborne Particles in Dust-Controlled
Areas and Clean Rooms Using Instruments Capable of
Detecting Single Sub-Micrometre and Larger Particles
(Not ISO but Helps and is Current)

►For particles in the size range 0.1 µm

to 2 µm in diameter and a Single Particle
Counting Device flow rate of 0.028
m³/min (1 ft³/min), a transit tube up to 30
m long can be used.

►For particles size range 2 µm to 10 µm,

a maximum transit tube length of 3 m
can be used.
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Particle Sampling Tubing

ASTM F50: (Not ISO but Helps)
►If a flexible transit tube is to be used, then no
radius of curvature below 15 cm shall be used.

Probe With
Curvature of 15
CM Radius

Probe With too

Tight of a
Curvature
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PARTICLE SAMPLING TUBING

New ISO 1644-1:2015 Annex C (Informative not

Mandatory)-- Counting and sizing of airborne
macroparticles states:

Clause C.4.1.2 “For sampling of particles larger

than and equal to 1 μm, the transit tube length
should not exceed the manufacturer’s
recommended length and diameter, and will
typically be no longer than 1 metre in length.”

FAQs ON PARTICLE
COUNTING
How Often Should we
Classify an ISO Class 5
cleanroom?

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FREQUENCY OF CLASSIFICATION OF
CLEANROOMS
• At-rest and operational classification should performed
periodically based upon risk assessment of the cleanroom
and clean zone operations, typically on an annual basis
• Where the cleanroom and clean zone is equipped with
instrumentation for continuous or frequent monitoring of air
cleanliness (airborne particles, room pressure differentials),
the time intervals between classification may be extended
provided that the results of the monitoring remain within the
specified limits. However, in the pharmaceutical and
related industries formal classification / re-qualification
must be undertaken at least annually
• Frequency of re-testing cleanroom no longer dictated by
Class, to be done annually. Previously it was 6 months for
ISO Class 5

FAQs ON PARTICLE
COUNTING
How Should we
conduct the Recovery
Test?

Revision to Room Recovery Rates

ISO/DIS 14644-3 Section B.4:

•For non-unidirectional cleanroom only

•This test now incorporates 100:1 and 10:1 tests.

•Test should be performed when the clean room is in the ‘As Built’
or ‘At Rest’ states, or after major modification to the
cleanroom, or its operation.

•No requirement to retest every 2 years.

•It is not recommended that the 100:1 test be used for ISO 8 & 9

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RECOVERY TEST according to new

ISO/DIS 14644-3:2016
Clause B.4 Recovery Test (previously Clause B.12)
This test is performed to determine the ability of the
installation to reduce the concentration of airborne
particles by dilution. (Previously clause said This test is
performed to determine the ability of the installation to
eliminate airborne particles.)
Recovery performance is evaluated by using the 100:1 or
10:1 recovery time and/or the cleanliness recovery rate.
The 100:1 or 10:1 recovery time is defined as the time
required for decreasing the initial concentration by a factor
of 100 times (and 10 times).
Previously only 100:1 test was mentioned, now 10:1
also.

RECOVERY TEST
. Where to test:

•Working level
•At places where particle concentration indicates further
investigation
•At product / process level
Do not test at:
•Places where there is Direct influence of air supplies
•Near air returns

When an artificial aerosol is used, the risk of residue

contamination of the installation should be considered.

RECOVERY TEST- PROCEDURE

• Care should be taken to avoid coincidence error and
potential contamination of the Particle Counter optics.
Before testing, calculate the concentration required to
carry out the recovery test. If the concentration exceeds
the maximum capability of the Particle Counter such that
coincidence loss occurs: 1) use the dilution system; or 2)
reduce the concentration to avoid coincidence losses.
• The particle size used in this test should be not greater
than 0.5 μm. The cleanroom area to be examined should
be contaminated with an aerosol while the air -handling
units are in operation
• Raise the initial particle concentration to more than 10 or
100 times depending on the target cleanliness level
• Commence measurements at not more than 1 min
intervals and record time and concentration.

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Evaluation by 10:1 or 100:1

recovery time
Evaluation procedure:
a) note the time when the particle concentration reaches
the 10 or 100 ⨯ target concentration threshold (t10n or
t100n)
b) note the time and concentration when the particle
concentration reaches the target cleanliness level,
(tn)
c) the 10:1 recovery time is represented by t0.1 = (tn - t10n);
d) the 100:1 recovery time is represented by t0.01 = (tn -
t100n).

• Any Questions about the

Questions?

• Any Questions about the Answers?

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