Cleanrooms:

Classification versus Monitoring;

Considerations of Removal Efficiency
Setting Alerts and Actions
 Agenda

• Classification versus Monitoring

• What affects Particle Concentration in a cleanroom

• What can be readily controlled to affect concentration

• What are the current trends and dialog that will likely
change historic mainstays of cleanroom design

• Setting ALERT and ACTION levels for best effect

2
 Conclusion

Two key thoughts:

1. Know and control sources of contamination

2. Understand how the area/room removes contamination

3
 Sources of contamination

Sources:
In-filtration
Filtered air
Machines
Processes
Materials
People
- How many
- Gowning level and execution

4
 Removal Efficiency

• Quantity of filtered air

• Quality of filtered air
• Turbulence
• Impact of convection
• Cleaning routines

5
WWW.ASHRAE.ORG

USD 151.00

(as PDF)
 Measuring Particles: 2 intentions

Classification Monitoring
12
 Differences

Classification Monitoring
6 months or annual; Daily, weekly, monthly
Frequency
a formal study or continuous

Number of positions By formula By need for data

Sample volume By formula By need for data

By table; By need for trend info

Pass/Fail criteria or control; often
one “class limit” value ALERT and ACTION
In form needed for
Reporting format By standard
rapid understanding
Distribution of counts Uniform or Unique at each
in a room or zone homogeneous sample position
13
 Differences

Classification Monitoring

Focus of assessment Room or Zone Each sample position

14
 Measuring Particles: 2 intentions

Classification
15
 Classification Standards for Airborne Particles

General Cleanroom Airborne Particle Monitoring Standards

– before 1999

16
 Classification Standards for Airborne Particles

General Cleanroom Airborne Particle 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
1999 compliance with ISO 14644-1

ISO 14644

17
 Classification Standard: ISO 14644-1
General Standard for all Industries

Electronics
• Semiconductor
• Flat Panel
• Circuit Board
• Optical
• MEMS/Nanomachines
Life Sciences
Other
• Pharmaceutical
• Biotechnology Laboratory Electronics
• Medical Devices
• Hospitals/Pharmacies
Aerospace
• Launch Vehicles Aerospace
• Satellites
• Commercial/Military Aircraft
Laboratories Life Sciences
• Analytical Laboratories
• Universities
Other
• Nuclear
• Photographic, X-ray films
• Automobile Painting

18
 Classification Standard:
ISO 14644-1:1999

Purpose

• Defines cleanroom classes

• Establishes minimum sampling volume

– 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 a representative sample of the total air volume from a
statistically valid number of locations

19
 Classification Standard: ISO 14644-1:1999
Limits

Class Number of Particles per Cubic Meter by Micrometer Size

0.1 µm 0.2 µm 0.3 µm 0.5 µm 1 µm 5 µm

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

FS 209E
ISO 5 100,000 23,700 10,200 3,520 832 29
Class 100

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

FS 209E
ISO 7 352,000 83,200 2,930
Class 10,000
FS 209E
ISO 8 3,520,000 832,000 29,300
Class 100,000

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

20
 Revised Table for ISO Classes;
Classification Limits: ISO 14644-1:2015

Number of Particles per Cubic Meter by Micrometer Size

ISO
0.1 µm 0.2 µm 0.3 µm 0.5 µm 1 µm 5 µm

1 10

2 100 24 10

3 1,000 237 102 35

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

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

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

7 352,000 83,200 2,930

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

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

21
Revisions to ISO 14644-1

( December 2015 )
 Major change #1:

Minimum number of sample

locations
 Revisions to ISO 14644-1

Method of determining minimum number of

sample positions
– Previously determined by taking the Square Root
(SQRT) of measurement area (in square meters)
– Replaced with stated number of minimum sample
positions as a look-up chart
– Will mean a modest increase in the number of
sample points in almost all cases

24
 Table A.1 — Sample locations related to cleanroom area

Area of cleanroom (m2) less Minimum number of sample

A.4.1 Establishment of than or equal to locations to be tested (NL)

sampling locations 2 1

4 2

6 3

8 4

Derive the minimum number 10 5

24 6
of sampling locations, NL, 28 7

from Table A.1. 32

36
8

9
52 10
Table A.1 provides the 56 11
64
number of sample locations 68
12

13

related to the area of each 72 14

76 15
cleanroom or clean zone to 104 16

be classified and provides at 108

116
17

18
least 95 % confidence that 148 19

at least 90 % of all locations 156

192
20

21
do not exceed the class 232 22

limits. 276

352
23

24
436 25
636 26
1000 27
> 1000 See Equation A.1
25
 Major change #2:

Specific Calibration Method

( ISO 21501-4:1997 )
 ISO 21501-4:
Additional Tests

Basic calibration ISO 21501-4

• Size calibration • Size calibration
• False count rate • False count rate
• Sampling Flow Rate • Sampling flow rate
• Sampling Time • Sampling time
• Verification of size setting
• Counting efficiency
• Size resolution
• Concentration limit
• Sampling volume

27
 Repeatability

Measurement Measurement Measurement

#1 #2 #3

28
 Repeatability

6 months 12 months

29
Reproducibility

30
 Table A.1 — Sample locations related to cleanroom area

Area of cleanroom (m2) less Minimum number of sample

A.4.1 Establishment of than or equal to locations to be tested (NL)

sampling locations 2 1

4 2

6 3

8 4

Derive the minimum number 10 5

24 6
of sampling locations, NL, 28 7

from Table A.1. 32

36
8

9
52 10
Table A.1 provides the 56 11
64
number of sample locations 68
12

13

related to the area of each 72 14

76 15
cleanroom or clean zone to 104 16

be classified and provides at 108

116
17

18
least 95 % confidence that 148 19

at least 90 % of all locations 156

192
20

21
do not exceed the class 232 22

limits. 276

352
23

24
436 25
636 26
1000 27
> 1000 See Equation A.1
32
 Intuitive User Interface!

Test wizard for standards

compliance

• Wizard to conduct pass/fail test for:

– ISO 14644-1
– EU-GMP Annex I
– FS 209E standards
• No expertise in standards required.
Just a few clicks to compliance
• The wizard guides an operator
step by step to sample data,
process data and product reports

33
2 No more manual
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Files exported to Excel straight from the
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34
3 Built in workflow tools

MET ONE Simply Paperless:

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35
 5 Easy integration into LIMS

3. Data transferred
automatically via your network
in .pdf, .csv and .xml formats
1. Take 2. Click ‘Export’
Options:
particle
a) Retain .pdf, .csv and .xml
count or
sample a) Feed data direct into LIMS

Accurate, 21CFR part 11

compliant data transfer!

MET ONE Simply Paperless increases EM

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36
MET ONE Simply Paperless 2 No more manual data entry
Summary
1
No more scanning printouts

Eliminate manual data transcriptions

3 Built-in workflow tools

Eliminate paper print-outs and scanning/photocopying

Step-by-step directions, eliminating data gaps. Includes

location labels, sample recipes, sample review/commen

5
Option of full LIMS integration
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Save up to 1½ hours per day

Fully electronic records

37
 Impact on

- EU GMP Annex 1
- PIC/s EU GMP Annex 1
- DR Norm 32
 None directly !!!

But …

because these reference ISO 14644-1

to determine the minimum number of
sample points, there is an effect . . .
 EU Annex 1:
Latest revision (2009)

Classification
– Sections 4 through 7

Monitoring
– Sections 8 through 17

40
 EU Annex 1 Summary:
Classification

Classification – Sections 4 through 7

Section 4:
“Classification should be clearly differentiated from
operational process environmental monitoring.”

Section 5:
“ For classification purposes in Grade A zones, a minimum
sample volume of 1 m3 should be taken per sample
position.

41
 EU Grade Definitions

at rest in operation
maximum permitted number of particles/m3 equal to or above
Grade Activity 0.5 µm 5 µm 0.5 µm 5 µm
A High Risk - filling, open vials, stopper bowls 3 520 20 3 520 20
B Aseptic preparations 3 520 29 352 000 2 000
C Clean area of less critical operations 352 000 2 000 3 520 000 20 000
D Clean area of less critical operations 3 520 000 20 000 not defined not defined

• Zone grades according to risk of product contamination

• Particle count measurements at 0.5 µm and 5 µm
• “At rest” vs “In operation”

42
 EU Annex 1:
Latest revision (2009)
Limits at 5 microns for Grade A
1 per cubic meter  20 per cubic meter

At Rest
At Rest In Operation
3
permitted number of particles/m3
Maximum permitted
Maximum
Grade
equal to
equal to or
or greater
greater than
than the
the tabulated
tabulated size
size
0.5 µm
0.5 µm 5 µm
5 µm 0.5 µm
0.5 µm 5 µm
5 µm
A
A 3 500
3 520 20
1 3 500
3 520 20
1
B 3 500
520 29
1 352 000
350 2 000
900

C 352 000
350 2 000
900 3 500
520 000 29 000
20
D 335000
520 000
000 29 000
20 not defined not defined

43
 EU Annex 1 Summary:
Classification

Section 5:

“For classification purposes EN/ISO 14644-1 methodology

defines both the minimum number of sample locations
and the [minimum] sample size based on the class limit
of the largest considered particle size and the method of
evaluation of the data collected.”

44
 EU Annex 1 Summary:
Classification

Section 5 (continued)
“For classification purposes EN/ISO 14644-1 methodology
defines both the minimum number of sample locations
and the [minimum] sample size based on the class limit
of the largest considered particle size and the method of
evaluation of the data collected.”

Number of locations Sample Volume (B,C,D)

• Based on lookup table

45
 EU Annex 1 Summary:
Monitoring
Monitoring: Sections 8 through 17
Section 8:
“Clean rooms and clean air devices should be routinely
monitored in operation and the monitoring locations
based on
– a formal risk analysis study
and
– the results obtained during the classification of rooms and/or
clean devices”

46
 EU Annex 1 Summary:
Monitoring
Section 9
• “The Grade A zone should be monitored at such a
frequency and with suitable sample size that all
interventions, transient events and any system
deterioration would be captured and alarms triggered if
alert limits are exceeded.

= “continuous” !!!

47
 EU Annex 1 Summary:
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.
48
 Monitoring Positions:
Risk-based Approach

Vial
Sterilizing 1 Lyo 1
5
Tunnel
7

2 3

Lyo 2
• Monitoring must follow the workflow, covering areas
where product is exposed – Annex 1 (2009)
− Where open vials exit de-pyrogenation – human interaction (1) 6

− Where vials are filled (2,3)

− Surrounding Grade B background (4)
─ Where the vials are partially stoppered (5) Lyo 3
─ Loading area in front of lyophilizers must be Grade A if
product is not fully stoppered (6,7)

49
 Monitoring Positions:
Risk-based Approach

Vial
Washing 1

System

2 3

In a filling operation for which the

final product remains liquid, some
points established for a lyophilized
product would not be needed.

50
 Measuring Particles: 2 intentions

Classification: Monitoring:
ISO 14644-1; Annex 1 Your SOP
51
 Key Points about Cleanrooms

Cleanrooms are dynamic; particle concentrations change

with location and with time
Particle counts can fluctuate considerably but yet be normal
Studying the actual particle count values over a long period
of time is often critical to setting good values for ALERT and
ACTION levels
Strategies for setting ALERT and ACTION levels will likely
be different for continuous PROCESS control versus
intermittent EM sampling

52
What affects Particle Concentration in a
cleanroom ???

53
 Cleanrooms and Clean Zones

“Cleanrooms and associated controlled environments

provide for the control of contamination of air or surfaces to
levels appropriate for accomplishing contamination
sensitive activities. Contamination control can be beneficial
for protection of product or process integrity in applications
such as the aerospace, microelectronics, pharmaceuticals,
medical devices, healthcare, food, etc.”

ISO 14644-1

54
 Cleanrooms and Clean Zones

3.1.1 cleanroom

Room within which the number concentration of airborne

particles is controlled and classified, and which is designed,
constructed and operated in a manner to control the
introduction, generation, and retention of particles inside the
room.

ISO 14644-1

55
 What can go wrong?
A cleanroom or cleanzone usually starts out clean
• What are my potential sources of contamination?
– How can I eliminate, minimize or control them?

• How does my room or zone remove particles?

– In what direction(s)?
– How fast?
– How many occupants can be in there at one time?
• Is my cleaning service effective?
• How can I test or monitor to know I’m OK?
 The equation

Particle
generation
Rate Expected
Removal Counts per
----------------- x =
efficiency volume
Dilution
rate
 Particle Generation: Sources

Filtered Air
Leakage into Room
Machinery
People
Deposition > re-circulation

58
 What can go wrong?
Do you know
which way the
wind blows?

Particle events !!
 Key Take-away #1

Particle concentration varies by:

A) Location

B) Time

62
63
Effect of Unidirectional Air Control

64
 FDA on smoke studies of interventions

Company X

Company
X

Company
X

65
FDA’s Inspectional Observations (483’s) on Air Flow Pattern Visualization

1. Smoke studies in ISO 5 hoods were not conducted under dynamic conditions.

2. There has been no air flow pattern (i.e smoke study) evaluation study
performed to determine the acceptability of the horizontal air flow, that is,
the air flow is not compromised (i.e air turbulence/air eddies) during
the aseptic operations that are performed in the ISO-5 area.

3. There has been no air flow pattern evaluation to determine that

the personnel activities and manual transfer of materials between the
ISO-8 and ISO-7 areas negatively affect the air movement and air cascade.

4. Smoke studies have not been properly documented for the air flow patterns
of the ISO 6 class rooms or ISO 5 laminar air flow hoods used in the processing
of injectable products.

66
FDA’s Inspectional Observations (483’s) on Air Flow Pattern Visualization

5. The air flow pattern video does not present data to adequately assess
the requested “downward sweeping air flow pattern” for the ISO 5
aseptic fill zone. The firm failed to evaluate the potential product impact
of the turbulence, air eddies observed in the middle of the ISO 5 hoods
during dynamic operations.

6. Smoke study did not include an evaluation of the personnel activities

performed in the adjacent ISO 5 hoods to determine that the personnel
activities do not negatively affect air flow patterns within ISO 5 hoods.

7. The smoke study does not demonstrate critical aseptic connections

performed during the assembly of filling equipment used to fill
sterile pharmaceuticals within ISO 5 hoods

67
 Smoke tests

Airflow Visualization Techniques and

Practices - August
Aug 15 - Aug 17, 2017 / Bethesda, MD

68
 People contribute particles

What
74
 People contribute particles

Study into Human Particle Shedding, Cleanroom Technology, August 2011, pages 26- 28
 The equation

Particle
generation
Rate Expected
Removal Counts per
----------------- x =
efficiency volume
Dilution
rate
 General Air Monitoring

• Non-viable counts
– Sometimes referred to as “total count”
– Includes all types of airborne material
• Solid particles
• Fibers
• Microorganisms
• Skin flakes
• Droplets
Table salt Bacteria
40 µm

Skin flake
What can be readily controlled ???

80
 What can be controlled

• Fan Speed/Air Change Rate

• Number of sources and source strength
• Coverage of Garments
• Quality of garments/frequency of
washing/lifetime
• Activity of personnel
• Workflow and Position in room

81
82
 Study of people as sources in ISO 5

Contamination of cleanrooms by people, John Sharp et al,

European Journal of Parenteral & Pharmaceutical Sciences 2010; 15(3): 5-11
84
 Study of people as sources in ISO 5

Contamination of cleanrooms by people, John Sharp et al,

European Journal of Parenteral & Pharmaceutical Sciences 2010; 15(3): 5-11
85
What are the current trends and dialog that
will likely change historic mainstays of
cleanroom design ???

86
 Current trends and topics

• Impact of Isolators and RABS

• Robots
• Energy Conservation
• Historic guidelines for Flow Rates and Air change Rates

87
 Why reduce air flow/air change rates ?

• Lower air change rates result in smaller fans, which

reduce both initial investment and construction cost.

• Fan power is proportional to the cube of air change rates

or airflow. A 30-percent reduction in air change rate
results in a power reduction of approximately 66 percent.

• By minimizing turbulence, lower airflow may improve

cleanliness.

88
Energy conservation – idle times

• If no sources are generating

particles (machinery or
people), why not turn down
the fan speed to save
energy?
• Still maintain room
characteristics of temperature
and humidity as needed
• Need to understand time
needed to bring room back to
desired levels for active use

90
Cleanroom???

91
 Cleanroom airborne contamination
control

• Control of people emissions – good garments are the primary control

measure
• Positive pressure to ensure external contamination is excluded
• well-sealed and cleanable cleanroom envelope to minimise
uncontrolled leak paths
• Correctly positioned, integral terminal H14 HEPA filters and housings to
supply particle free (no particles ≥0.5µm) air to the room
• Good ventilation effectiveness to ensure clean supply air is providing
good dilution and removal of contamination
• Cleanroom supply diffusers to effectively distribute clean air into the
room
• Low level extraction to help flushing away contamination
• A HVAC system to supply and extract sufficient contamination free
airflow to dilute and remove residual contamination from the room

92
 What does “sufficient airflow”
mean?

It depends who you ask……

Some companies (customers & designers) still talk about airflow in terms of
needing a certain minimum number of air-changes per hour (ac/hr) to meet
their cleanliness requirements….

93
Typical ac/hr across industry

94
 Air-change rate as a basis for design
leads to..

• Unpredictable cleanliness levels

• Cleanrooms which are much cleaner than they are required to be
• Possibly less clean operationally than required
• Facilities which are much larger than necessary (AHU / Ducts etc)
• Facilities which are more costly to build than necessary
• High HVAC energy use
• Greater emissions
• Greater impact on the environment
• Less sustainable
• Sub-optimised

95
 What’s the answer?

Follow the science, consider HVAC design based on a Scientific

approach rather than ad-hoc air-changes to ensure an optimised and
transparent solution is delivered.

96
 A scientific approach
to determine airflow

Determine emissions into the room (D), agree required level of cleanliness in
the room (C), and calculate the amount of supply air required (considering
ventilation effectiveness)
97
100
Removal or Ventilation Efficiency

101
Monitoring Compressed Air/Gas
for Particles

1
FDA: Air (CDA) or Gas
that contacts Product

103
 Air (CDA) or Gas
that contacts Product
Frequency of sampling and target levels for CDA/Gases

It would seem that the authorities have not established a periodicity for
sampling of compressed gas sources. In cases where there is concern
that there might be some risk, it may be wise to sample gas sources on
a quarterly basis and to set an expectation of an ISO 7 level in non-
sterile applications; for sterile areas, a target of ISO 5 or better should
be used. Many customers look for an ISO 4 level in sterile gas supplies
and this should be achievable with most commercially available filter
methods. This higher target level is, however, one of choice, rather than
one dictated by regulation (but does add some safety margin).

Also, for sterile areas, a more frequent - monthly or even weekly -

sampling may help minimize any lengthy period of heightened risk due
to a system failure.

104
 MET ONE 3400 for CDA/Gas testing

Ordering a Particle Counter for Gas Sampling

The new 3400 gas option, together with the
appropriate High Pressure Diffuser (HPD), makes gas
monitoring straight-forward. The user simply selects
the required gas from a pull-down list on the 3400
interface; the sample flow is automatically adjusted to
ensure that a calibrated flow rate is maintained to
specification.
Ordering a new 1 CFM 3400 with gas calibration is likewise
straight-forward. Four versions are available, and include gas
calibrations for these gases: Air, N2, and CO2.

2088900-06 3413 0.3 micron, 1 CFM, GASES

2088900-07 3413 0.3 micron, 1 CFM, GASES, WIFI
2088900-08 3415 0.5 micron, 1 CFM, GASES
2088900-09 3415 0.5 micron, 1 CFM, GASES, WIFI
For the 1 CFM counters listed above, select the 2080732-13 HPD. Note that HPD
versions are available for other flow rates when CDA and/or N2 are the only gases
105
to be sampled.
MET ONE Diffusers for CDA/Gas

106
 Summary

Cleanrooms are dynamic; particle concentrations change

with location and with time
Particle counts can fluctuate considerably but yet be normal
Studying the actual particle count values over a long period
of time is often critical to setting good values for ALERT and
ACTION levels
Strategies for setting ALERT and ACTION levels will likely
be different for continuous PROCESS control versus
intermittent EM sampling

107
 What can go wrong?
A cleanroom or cleanzone usually starts out clean
• What are my potential sources of contamination?
– How can I eliminate, minimize or control them?

• How does my room or zone remove particles?

– In what direction(s)?
– How fast?
– How many occupants can be in there at one time?
• Is my cleaning service effective?
• How can I test or monitor to know I’m OK?
 Conclusion

Two key thoughts:

1. Know and control sources of contamination

2. Understand how the area/room removes contamination

109
Thank you !

joegecsey@gmail.com

+1 541 295 7538

Presented by

Joe Gecsey
Grants Pass, Oregon USA
Life Science
Applications

110