Particle Counting

-
Theory
-
Guidelines
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Monitoring
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Liquid Particle counters

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 Agenda
• Particle counting – Introduction

• Optimizing Sampling of Airborne Particles

• Recent and Pending Changes concerning Airborne Particle Counting ISO

14644-1, Annex 1, ISO 21501

• Integration and Automation – Getting It All Together: Industry Trends & Case
Studies

• Integration and Automation – Getting It All Together: Defining your

Requirements

• Particles in Liquids: Sampling Techniques and “How to’s”; Satisfying USP

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 Recent and Pending Changes
concerning Airborne Particle
Counting ISO 14644-1, Annex 1,
ISO 21501

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 Classification Standards for Airborne Particles

General Air Monitoring Standards

– ISO 14644-1
• Classification of air cleanliness
– ISO 14644-2
• Specifications for testing and monitoring
to prove continued compliance with ISO
14644-1
– ISO 14644-3
• Guidance on instrumentation to be used
for testing for compliance with ISO 14644-
1999 1

ISO 14644 4
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 Classification Standard
ISO 14644-1
• Defines cleanroom classes in a zone

• Establishes minimum sampling volumes

– Purpose: Gather a sample volume with theoretically
at least 20 particles for a statistically valid sample

• Establishes minimum number of points to classify area, based on

statistical criteria
– Gather from a valid number of locations for a representative sample of
the total air volume

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 Classification: ISO 14644-1

A sample size with at least 20 theoretical particles, or

Minimum sample volume 2.0 liter

Minimum sample time 1 minute

Minimum number of 1
whichever is
locations greatest
with at least 3 samples
total

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 Classification Limits ISO 14644-1

Class Number of Particles per Cubic Meter by Micrometer Size

0.1 m 0.2 m 0.3 m 0.5 m 1 m 5 m

ISO 1 10 2

ISO 2 100 24 10 4

ISO 3 1,000 237 102 35 8

ISO 4 10,000 2,370 1,020 352 83

ISO 5 100,000 23,700 10,200 3,520 832 29

ISO 6 1,000,000 237,000 102,000 35,200 8,320 293

ISO 7 352,000 83,200 2,930

ISO 8 3,520,000 832,000 29,300

ISO 9 35,200,000 8,320,000 293,000

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 What volume do I need to collect?

• ISO 14644 requires sample with 20 theoretical particles

• ISO 14644 allows 3520 particles (0.5um) in 1m3 (1000 liters)

20 particles
Volume = X 1000 liters
3520 particles

Volume = 5.7 liters

Greater than 2.0 liters so minimum sample volume is satisfied

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 ISO 14644-1 Minimum Sample Time at 1 CFM

Time required (in minutes) at 1 cfm (28.3 lpm) flow rate with 1-minute limit imposed

0.1 um 0.2 um 0.3 um 0.5 um 1 um 5 um

ISO Class 1 70.64 353.20

ISO Class 2 7.06 29.43 70.64 176.60

ISO Class 3 1.00 2.98 6.93 20.18 88.30

ISO Class 4 1.00 1.00 1.00 2.01 8.51

ISO Class 5 1.00 1.00 1.00 1.00 1.00 24.36

ISO Class 6 1.00 1.00 1.00 1.00 1.00 2.41

ISO Class 7 1.00 1.00 1.00

ISO Class 8 1.00 1.00 1.00

ISO Class 9 1.00 1.00 1.00

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 Continued Compliance:
ISO 14644-2
Schedule of Tests to Demonstrate Continuing Compliance

Maximum Time
Test Parameter Class Test Procedure
Interval

Particle Count
≤ ISO 5
All 24
6 Months
ISO 14644-1
Test classes
> ISO 5 12months
Months
Annex A

Important
Air Pressure note: If Particle counts and Differential
ISO 14644-1
All Classes 12 Months
Pressure
Difference are monitored continuously (forAnnex B5
example, by a Facility Monitoring System) then
ISO 14644-1
the Maximum Time Interval for re-certification
Airflow All Classes 12 Months
Annex B4
can be extended
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 Example: ISO 14644-1 Classification

Vial
Washing Lyo 1
Station 5m

Lyo 2
8m

5m
Calculations for Number of Points:
Area of clean zone = 80 m²
Lyo 3
Take the SQRT (80) = 8.94
Rounding up to next integer = 9 sample positions

4m

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 Example: ISO 14644-1 Classification

Vial
Washing Lyo 1
Station
1 2 3 4 5 6

Lyo 2

Calculations for Number of Points:

Area of clean zone = 80 m²
Take the SQRT (80) = 8.94 9
Lyo 3
Rounding up to next integer = 9 sample positions

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 Example
ISO 14644-1 Calculations
1 2 3 4 5 6 7 8 9 10

Vial
Freeze
Washing
Dryer 1
System

Freeze
Dryer 2
• Need to adjust for equipment in room.
• Under ISO 14644-1, if you sample at 10 or more
positions, you can avoid the added calculation of
the UCL (Upper Confidence Limit). Calculation of
the UCL is only mandated when the number of
positions used is between 2 and 9. Freeze
• Best to sample near potential problem spots which Dryer 3
are near entrances and exits and near operator
positions.

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 Example
ISO 14644-1 Calculations

9
1
Vial 10 Freeze
Washing
8 Dryer 1
System
2

11
3 4 5 6 7

12 Freeze
Dryer 2
• Need to adjust for equipment in room.
• Under ISO 14644-1, if you sample at 10 or more 13
positions, you can avoid the added calculation of
the UCL (Upper Confidence Limit). Calculation of
the UCL is only mandated when the number of
positions used is between 2 and 9. Freeze
14
• Best to sample near potential problem spots Dryer 3
which are near entrances and exits and near
operator positions.

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 Example
ISO 14644-1 Calculations

1. Average sample data values at each position

2. Normalize the average to number of particles per cubic meter
3. Compare normalized value to the target class limit; normalized value
at each and every sample point must be less than the limit for the
given size and target room classification

If the number of points sampled is more than 1 but less than 10,
then the UCL factor must be applied:
a) Calculate the standard deviation
b) Use Student’s T-factor from tables
c) Calculate UCL
d) Compare to classification limit; UCL must
not exceed the applicable limit

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 Probable Revisions to
ISO 14644-1, -2
1. Frequency of re-certification
Now:
• If zone is ≤ ISO Class 5, every 6 months
• If zone is ≥ ISO Class 6, every 12 months

Proposed:
• For all zones, every 12 months

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 Probable Revisions to
ISO 14644-1, -2

2. Eliminate Student’s T test for sample plans with 1 to 9

sample positions

3. Method of determining number of sample positions

– Replace with stated number of minimum sample positions
as a look-up chart
– Based on 95% confidence levels not on SQRT of area
– May mean a small increase in the number of sample points

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 B.4.1.1

Derive the minimum number of

sampling locations NL from table 3.

Table 3 shows the number of sample

locations related to the area of each
cleanroom or clean zone to be
classified and provides at least 95%
confidence that at least 90 % of all
locations do not exceed the class
limits.

Different levels of confidence and

verification can be specified and
agreed upon by the customer and
supplier.

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 ISO
Room Room ISO 14644-1: FS209E FS209E FS209E FS209E
Size Size 14644-1: 2007 Unidirectional Turbulent Turbulent Turbulent
(ft2) (m2) 1999 (95,90) (all classes) Class 1000 Class 10K Class 100K
21.5 2 2 1 2 2 2 2
43.1 4 2 2 2 2 2 2
64.6 6 3 3 3 3 2 2
86.1 8 3 4 4 3 2 2
107.6 10 4 5 5 4 2 2
129.2 12 4 6 6 5 2 2
150.7 14 4 7 7 5 2 2
172.2 16 4 8 7 6 2 2
193.8 18 5 9 8 7 2 2
279.9 26 6 10 12 9 3 2
301.4 28 6 11 13 10 4 2
344.4 32 6 12 14 11 4 2
366.0 34 6 13 15 12 4 2
387.5 36 6 14 16 13 4 2
409.0 38 7 15 17 13 5 2
559.7 52 8 16 23 18 6 2
581.3 54 8 17 24 19 6 2
624.3 58 8 18 25 20 7 2
796.5 74 9 19 32 26 8 3
839.6 78 9 20 34 27 9 3
1033.3 96 10 21 42 33 11 4
1248.6 116 11 22 50 40 13 4
1485.4 138 12 23 60 47 15 5
1894.4 176 14 24 76 60 19 6
2152.8 200 15 25 87 69 22 7
3229.2 300 18 26 130 103 33 11
5382.0 500 23 27 216 171 54 18 19
10763.9
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5000 71 29 2153 1702 539 171
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Particle107639.1
Counting 10000 100 29 4306 3404 1077 341
 Probable Revisions to ISO 14644-1, -2
4. Remove possibility to classify
at 5 micron for ISO Class 5
Limit number of 29 removed
“Note (b) Due to sampling difficulties affecting collection of large
particles in low concentrations, specification of sampling by DPC
is inappropriate for classification at the indicated class.

5. Indicate that single digit limits for ISO Class 1 and 2

create challenges to timely execution

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 Probable Revisions to ISO 14644-1, -2
6. Normative Reference to ISO 21501-4

Normative reference
The following normative document contains provisions, which, through reference in
this text, constitute provisions of this part of ISO 14644. For dated references,
subsequent amendments to, or revisions of, any of these publications do not apply.
However, parties to agreements based on this part of ISO 14644 are encouraged to
investigate the possibility of applying the most recent editions of the normative
documents indicated below. For undated references, the latest edition of the
normative document referred to applies. Members of ISO and IEC maintain registers
of currently valid International Standards.

ISO 21501-4:2007 Determination of particle size distribution – Single particle light

interaction methods-Part 4: Light scattering airborne particle counter for clean spaces.

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 What is ISO 21501?

• ISO 21501 is a new family of standards describing the

instruments and calibration requirements for determining
particle size distribution using light interaction methods for
both liquid and airborne particle counters.

• ISO 21501 represents the culmination of work by

instrumentation manufacturers and industry users and comes
at a critical time for the life science industry with the increasing
trend for real-time air particle monitoring in cleanrooms using
light scattering air particle counters.

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 What is ISO 21501 replacing?
Previous calibration methods guidelines:
• ASTM F 328-98(2003) “Standard Practice for Calibration of an
Airborne Particle Counter Using Monodisperse Spherical Particles”
(withdrawn May 2007).
• IEST-RP-CC014.1 “Calibration and Characterization of Optical
Airborne Particle Counters” (providing actual methods to perform the
calibration).
• JIS B 9921:1997 - Japanese standard which comprehensively deals
with OPC design performance, most notably in the area of counting
efficiency.

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 ISO 21501 Additional Tests

Before ISO 21501-4 ISO 21501-4

• Size calibration • Size calibration
• Verification of size setting
• Size resolution • Counting efficiency
• False count rate • Size resolution
• False count rate
• Concentration limit
• Sampling time
• Sampling flow rate
• Sampling time
• Sampling volume 24
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 ISO 21501 Requirements
ISO 21501 Requirements
21501-2 21501-3 21501-4
Parameter Comments
LS Liquid LE Liquid Airborne
Standard uncertainty of the mean size
Size calibration +/- 2.5% +/- 2.5% +/- 2.5%
Median voltage
+/- 15% +/- 10%
+/- 10%
Verification of size setting Cal curve Cal curve Use 10% (As-received)
Reference material
(minimum = 3 points) (minimum = 3 points)
Close to the minimum detectable size.
50% Counting efficiency 50 +/- 30% N/A 50 +/- 20%
Concentration < 25% of concentration limit.
100 +/- 30% 100 +/- 20% 100 +/- 10%
100% Counting efficiency 1.5 to 3X minimum Reference material 1.5 to 2X minimum Use 1.5 to 2X minimum size
size < 2X minimum size size
Manufacturer's recommended particle size.
Use area under the curve.
Size resolution 10% 10% 15%
>1.5X
CC sets markers at 5% medium value (counts)
Particles per liter Particles per cubic Poisson distribution with a 95% confidence limit
False count rate N/A
(minimum size) meter (minimum size) On screen calculator

Maximum particle Particles per cubic Particles per cubic Particles per cubic 10% Coincidence loss (calculated)
concentration centimeter centimeter meter (minimum size) On screen calculator
Volumetric flow rate
MFR standard MFR standard
Sample flow rate +/- 5% If unit has flow rate control system
uncertainty uncertainty
Include flow rate uncertainty (all)
Sample time +/- 1% +/- 1% +/- 1% On screen calculator
Sample volume +/- 5% +/- 5% N/A On screen calculator
10 min at concentration limit (minimum size),
Response rate N/A N/A 0.5% Sample (<60 sec) then filter (10 sec), then
sample (< 60 sec)

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 Questions?

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