Understanding ISO 14644-1/2 2015

and Becoming Compliant

Daniele Pandolfi
Global Product Line Manager
Particle Measuring Systems
dpandolfi@pmeasuring.com
 Agenda
• What’s the ISO 14644
• ISO 14644-1:2015 Cleanroom certification revision summary
• Notable areas of changes
• ISO 21501-4 Instrument Implications
• ISO 14644-2:2015 Cleanroom Monitoring revision summary
• Monitoring Plan Alternatives
• Risk Assessment
• Alert and Action Limits
• How to chose the right strategy
• Learn from FDA 483 Warning Letters
Introduction
• The recent revision of ISO 14644-1 and-2 has
introduced several changes for cleanroom
classification and monitoring guidelines.
• The ISO community voted in favor of this revision
on October 29th, 2015.
• This presentation discusses those changes and
how they affect you.
• Questions will be addressed at the end, but feel
free to voice them as they come to mind.
 What is ISO 14644?
ISO 14644 consists of the following parts, under the general title Cleanrooms and associated
controlled environments:
• Part 1: Classification of air cleanliness by particle concentration
• Part 2: Monitoring to provide evidence of cleanroom performance related to air cleanliness by particle
concentration
• Part 3: Test methods
• Part 4: Design, construction and start-up
• Part 5: Operations
• Part 7: Separative devices (clean air hoods, gloveboxes, isolators and mini-environments)
• Part 8: Classification of air cleanliness by chemical concentration (ACC)
• Part 9: Classification of surface cleanliness by particle concentration
• Part 10: Classification of surface cleanliness by chemical concentration
 What Is ISO 14644-1?
ISO 14644 represents the most used standards in Pharma
and Electronics controlled environments.

• ISO 14644-1 specifies classes of air cleanliness in terms of the

number of particles expressed as a concentration in air
volume.
• It also specifies the standard method of testing to determine
cleanliness class, including selection of sampling locations.
 New 14644-1:2015 Revision and Purpose

Simplify the classification process and

remove the need to evaluate the 95%
upper confidence limit (UCL) for low
sample location numbers (currently
required for 2/9 of cleanroom
locations).

Over the last five years, the

Review the classification procedure
and make it more applicable to
cleanroom operation. For example,
contamination is not expected to
be evenly distributed.
ISO Technical Committee 209
Update the standard as required
has been working on the to current reasoning and
revision of the basic airborne industry requirements.
cleanliness classification,
14644-1 and -2.

Avoid any radical change to the

principles of the current ISO
cleanliness classes 1-9.
New 14644-1:2015 Revision Summary and Purpose

Notable areas of change:

• Sample Location Number
• Particle Concentration Limit
• Particle Counter Calibration
 New 14644-1:2015 – Sample Location

How to determine the sample location number:

• A new table has been developed for the determination of the number of
sample locations, replacing “𝐿𝑜𝑐𝑎𝑡𝑖𝑜𝑛 𝑁𝑢𝑚𝑏𝑒𝑟 = 𝑚2 𝑟𝑜𝑜𝑚 𝑎𝑟𝑒𝑎”
from the ISO 14644-1:1999 version of the standard.

• For all room sizes above 6 m2, the new table results in an increase of
required sample locations.

• The ISO14644-1:1999 standard required the UCL 95% (Upper Confidential

Limit) calculation for sample locations to be between 2 and 9.
 Area of zone [m2] ISO 14644-1:1999 ISO 14644-1:2015
New 14644-1:2015 Sample Location 2 2 1
Table 4
6
2
3
2
3
8 3 4
What this means to you: 10 4 5
24 5 6
• Sample location number 28 6 7
32 6 8
calculations(*) are now unnecessary. 36 6 9
52 8 10
• * For all Cleanrooms Smaller than 56 8 11
1000m2 use the table. 64 8 12
68 9 13
• * For all Cleanrooms Bigger than 72 9 14
76 9 15
1000m2 use Formula A.1. 104 11 16
108 11 17
116 11 18
Formula A.1: 148 13 19
𝐴 𝑚2 156 13 20
𝑁𝐿 = 27 × 192 14 21
1000 232 16 22
276 17 23
352 19 24
436 21 25
636 24 26
1000 32 27
>1000 n/a See Formula A. 1
 New 14644-1:2015 – Sample Location
• The new table has been pre-calculated to eliminate the
need for this calculation.
• The new method, when successfully applied, assures that
at least 90% of the room is compliant
at a 95% confidence limit.

The determination of each sampling location will be based on a

semi-random sampling technique, derived from "hypergeometric"
distribution, which is the statistical model for sampling without
replacement.
 ISO 14644-1:1999 – Maximum Concentration Limits
Table 1 Selected airborne particulate cleanliness classes

ISO 14644-1:1999 Maximum concentration limits (particles/m3)

Classification
Number (N) 0.1 µm 0.2 µm 0.3 µm 0.5 µm 1.0 µm 5.0 µm

ISO Class 1 10
ISO Class 2 100 24 10
ISO Class 3 1 000 237 102 35
ISO Class 4 10 000 2 370 1 020 352 83
ISO Class 5 100 000 23 700 10 200 3 520 832 29
ISO Class 6 1 000 000 237 000 102 000 35 200 8 320 298
ISO Class 7 352 000 83 200 2 930
ISO Class 8 3 520 000 832 000 29 300
ISO Class 9 35 200 000 8 320 000 293 000
 ISO 14644-1:2015 – New Maximum Concentration Limits
Table 1 Selected airborne particulate cleanliness classes

ISO 14644-1:2015 Maximum concentration limits (particles/m3)

Classification
Number (N) 0.1 µm 0.2 µm 0.3 µm 0.5 µm 1.0 µm 5.0 µm

ISO Class 1 10
ISO Class 2 100 24 10
ISO Class 3 1 000 237 102 35
ISO Class 4 10 000 2 370 1 020 352 83
ISO Class 5 100 000 23 700 10 200 3 520 832
ISO Class 6 1 000 000 237 000 102 000 35 200 8 320 298
ISO Class 7 352 000 83 200 2 930
ISO Class 8 3 520 000 832 000 29 300
ISO Class 9 35 200 000 8 320 000 293 000
 ISO 14644-1:2015 – New Maximum Concentration Limits

As shown on the previous slide, the foremost concern in the Life Science industry
is the removal of the ≥ 5 µm particle concentration in ISO Class 5 clean areas
(for classification purposes) when compared to the ISO 14644-1:1999 version

In the 1999 version, the limit is 29 particles per cubic meter.

This change to the ISO 14644 standard is a major concern for a number of
reviewers.
 ISO 14644-1:2015 – Dealing with ≥ 5.0 micron particles

Primary reasoning for the de-emphasis of the ≥ 5 µm ISO Class 5 limit include:

• Sampling and statistical limitations for particles in low

concentrations make classification inappropriate.
• In order to specify this particle size in association with ISO Class
5, the macroparticle descriptor ‘M’ may be adapted and used in
conjunction with at least one other particle size.
• Sample collection limitations for both particles in low
concentrations and sizes greater than 1 μm make classification
at this particle size inappropriate, due to potential particle losses
in the sampling system.
 ISO 14644-1:2015 – Concentration Limit Considerations

• The ISO 14644-1:2015 is now the

same as the old US Fed Std 209E in
the treatment of 5 micron particles
in ISO Class 5 (old FS 209E class
100).
• This change was largely influenced
by US expert argument.

5 µm PSLs on a cotton fiber

 New ISO 14644-1:2015 Instrument Calibration Requirements

14644-1:1999

14644-1:2015

What this means to you:

You are expected to have instruments calibrated in accordance with ISO requirements.
Use of non ISO21501-4 compliant instruments will require additional and
undesirable explanations to authorities.
 What Is ISO 21501-4?

• The ISO 21501-4 is the worldwide-

recognized standard for optical particle
counter calibration.
• This standard was introduced by the
ISO organization in 2007, and it is
based on the following purpose.
 ISO-21501 – Purpose

• “…to provide a calibration procedure and

verification method for particle counters, so as to
minimize the inaccuracy in the measurement result
by a counter, as well as the difference in the results
measured by different instruments.”
• Four parts: …-2 (LSLPC), -3 (LELPC), -4 (LSAPC)
(LSAPC: Light Scattering Airborne Particle Counter)
 ISO-21501 – Why Is It Important?

• The ISO 14644-1:2015 introduces the need of ISO 21501-4 particle

counters, which assure all cleanroom certification is based on
verified data accuracy and reliability.
• Cleanroom users shall then look to ISO 21501 as a method to meet
cGMP, EU GMP, ISO 14644-1, and other requirements.
• This change to the ISO 14644-1 represents an important step in
improving the accuracy of clean room contamination evaluation
and aseptic process control.
 ISO 14644-1:2015 – Concentration Limit Changes

• The new revision of ISO 14644-1 leaves a gap in

classification of Grade A and B (at rest) in Annex 1 of the EU
GMP, as well as PIC/S, WHO, Japanese and China GMP
guidance for sterile products. Limits are provided to be
between 20 and 29 parts/m3.
• To deal with this problem, ISO 14644-1:2015, Annex C7,
introduces the use of the macro-particle descriptor ‘M’,
which can be used to describe either the specified or
measured concentration of macroparticles ≥ 0.5 µm.
 ISO 14644-1:2015 – New Standard Implementation/Migration

• GMP guidance refers to ISO standards, making the

standards a part of GMP guidance.
• The 2008 Annex 1 of the EU GMP, and US FDA's 2004
Aseptic processing guidance refer to ISO 14644-1 and ISO
14644-2 without any specific date or revision. This means
the latest version always applies.
• It is up to the GMP authorities to inform applicable
parties of the grace period and implementation date of
the revised standards.
• Users of cleanrooms should have a clear policy of
migration to the new standards.
 Standards changes and technology feature

• The dynamism of modern industry could

potentially cause further upgrading of
standards in the near future.
• Misinterpreting changes to standards can
make implementation difficult.
• The use of modern measurement devices
can help prevent deviation, immediately
enabling usage of the most updated
requirements.

Particle Measuring Systems Lasair III

Particle Counter Statistic Sample Screen
Understanding ISO 14644-2:2015
 What Is ISO 14644-2?
ISO 14644 represents the most used standards in
Pharma
and Electronics controlled environments.
• ISO 14644-2 specifies the requirements of a monitoring plan based on
risk assessment of the intended use. The data obtained provides
evidence of cleanroom or clean zone performance related to air
cleanliness by particle concentration.
• This revision of ISO 14644-2 emphasizes the need to consider a
monitoring strategy in addition to the initial or periodic execution of the
classification of a cleanroom or clean zone in accordance with ISO
14644-1:2015.
 New 14644-2:2015 Main Goals

Emphasize the needs and

advantages of a planned cleanroom
contamination monitoring

Over the last five years, the

Enhance the installation and Provides the method for a
ISO Technical Committee 209
process knowledge, as to improve correct particles contamination
has been working on the
the risk assessment evaluation and alarm and warning limits
a faster reaction to any unexpected
revision of the basic airborne setting, based on a careful
cleanroom performance cleanliness classification, evaluation of data trends
14644-1 and -2.

Define the differences between a

simple periodic cleanroom
control and a more
intensive/complex monitoring
strategy
 New 14644-2:2015 Main Goals

ISO 14644-2:2015 shall not be only considered as a new standard with

which to be compliant, but mainly as beneficial tool to use in achieving
mature cleanroom environmental control.

Last but not least, the goal dictated by the TC 209 is:

Concretely reduce the operation cost by preventing

production loss
 ISO 14644-2:2015 - Terminology
MONITORING

• Monitoring is an observation of the

process made in accordance with a
specific method, able to provide clear
evidence of cleanroom performance.
• Monitoring can be “continuous”,
“sequential” or “periodic”.
 ISO 14644-2:2015 – Monitoring Plan

SEQUENTIAL Location
Location1
• Performed using sequential “n”…

multiplexing systems.
• Unaccepted methods for
Pharmaceutical Industry
• High risk of particle loss in Location5 Location2
long tubing while measuring
particles greater than 1µm

Location4 Location3
 ISO 14644-2:2015 – Monitoring Plan

CONTINUOUS
• Uses multiple particle counters,
one per individual location
• Continuous flow of data over Location A
time
• Immediate evaluation of
unexpected contamination
events
 ISO 14644-2:2015 – Monitoring Plan

PERIODIC Week 1
• Scheduled particle monitoring
frequency (i.e. once per week)
Week 2
• ISO 14644-2:2015 requires the
test frequency to be defined
and clearly specified Week 3

Week «n»
 ISO 14644-2:2015 – Sample Point
SAMPLE POINT LOCATION
• Understand the contamination
sources and their impact on the
activity in the cleanroom
– Locate particle counter probes as
close as possible to critical zones
Critical Point
• Maximum allowed distance
– 1 foot = 33cm
 ISO 14644-2:2015 – Instrument Selection
INSTRUMENT SELECTION
• Airborne particle collection efficiency,
suitability to monitor the selected
particle size(s), and accessibility for
maintenance, calibration, and repair
• Potential adverse impact of the
sampling system on the process
• Air sample flow rate and volume
– 1 cfm / 28.3 lpm commonly used
 ISO 14644-2:2015 – Instrument Selection – cont.
ISO REQUIREMENT
• ISO 14644-2:2015 requires that all instruments
employed in the cleanroom classification or
monitoring activities fully comply with ISO
21501-4.
• ISO 21501-4 states the calibration and
performance requirement for LSAPC (Light
Scattering Airborne Particle Counters)
 ISO 14644-2:2015 – Remote Sampling

HOW LONG CAN MY TUBING BE?

• The use of long sample transport tubes must be avoided,
when intending to evaluate particle concentrations at
sizes greater than 1 μm.
• ISO requires the adherence to maximum tubing lengths
as specified by the particle counter manufacturer, which
is typically between 1.5 and 2 meters. (ref. ISO 14644-1)
 ISO 14644-2:2015 – Choosing the right monitoring plan

• ISO 14644-2:2015 does not specifically provide

a link between the cleanliness class and the
recommended monitoring method.
• It is up to the user to choose the most
appropriate one based on their specific
manufacturing requirements and risk
assessment.
 ISO 14644-2:2015 – Choosing the right monitoring plan

• Life Science industries may consider other standards to

correctly and reasonably set-up their monitoring method
and frequency.
• World Health Organization(WHO) issued a document
titled Environmental Monitoring of Clean Rooms in
Vaccine Manufacturing Facilities in November 2012
which includes instruction on determining the best
monitoring frequency approach based on cleanliness
class.
ISO 14644-2:2015 – Choosing the right monitoring plan
 ISO 14644-2:2015 – The Risk Assessment
Risk Assessment Goals
• Correctly understand:
• The appropriate creation of
– the process
risk assessment
– the critical areas/locations
documentation is an
essential requirement for – possible sources of contamination
implementing a compliant – element/event that may
monitoring plan. compromise the cleanroom
performance, product quality and,
consequently, negatively affect the
operation cost
 ISO 14644-2:2015 – The Risk Assessment

ICH Q9 – QUALITY RISK ASSESSMENT

• The 2005 International Conference on Harmonisation of Technical Requirements for

Registration of Pharmaceuticals for Human Use is one of the best guidelines available for
proper risk assessment development, review and application.
 ISO 14644-2:2015 – The Risk Assessment
PDA - PARENTAL DRUG ASSOCIATION

• Fundamentals of an Environmental Monitoring Program,

Technical Report No.13 is also a useful document when
approaching monitoring plan development, and addresses
the needs of a meaningful, manageable and defensible
monitoring program.
 ISO 14644-2:2015 – The Risk Assessment
RISK ASSESTMENT ALTERNATIVE TOOLS

• Other useful tools to be considered for reliable risk

assessment development are:
– HACCP,
– FMEA/FMECA,
– PHA, FTA and HAZOP
 ISO 14644-2:2015 – The Risk Assessment

A responsible understanding of the production process and

installation performance aids in the:

• prevention of unexpected out

of specification conditions

• Support of power saving

activities
 ISO 14644-2:2015 - Alert and Action Limits
How to choose the right strategy

Action Level Alert Level

User set level at which, when User set level, defined to provide
exceeded, will require immediate early warning of a drift from normal
intervention, root cause conditions. This level should be used
investigation and corrective actions to prevent action level conditions.
 ISO 14644-2:2015 - Alert and Action Limits
How to choose the right strategy

• The right determination of action and alert limits is extremely important and must be supported by
the risk assessment and a consistent quantity of historical data.
 ISO 14644-2:2015 - Alert and Action Limits
How to choose the right strategy

• ISO 14644-2:2015 states the importance of a long term

evaluation as well as a yearly assessment of limits,
method and frequency.
• While not necessarily requiring a change, the assessment
is an important exercise in the critical evaluation of a
monitoring plan.
• Cleanroom users should frequently question whether
their existing monitoring plan is still applicable and
consistent with the cleanroom’s actual performance and
activities.
 ISO 14644-2:2015 - Alert and Action Limits
How to choose the right strategy

The standard provides some important recommendations, as well as an applicable strategy to

keep in mind when setting alert and action limits. One with high significance is provided in the
paragraph B.3.1.2, quoted below:

• B.3.1.3 When setting alert and action levels, it is important to be sensitive to the high variability of airborne particle concentrations with time and at
different locations. In particular, special care shall be taken when considering alert and action levels for cleanliness classes ISO Class 5 and cleaner
with low concentrations of particles. In these circumstances, the occurrence of “nuisance alarms” due to false counts and/or natural variability of
Frequent “nuisance”
particle concentration is more likely and should be avoided by careful selection of alert and action levels.
alarms should be avoided as they can lead to alarms being ignored by users.
 ISO 14644-2:2015 - Alert and Action Limits, How to choose the right strategy
STRATEGY 1
• Establish a trigger threshold value based on a series of consecutively high readings.
– For example: 3 consecutive, 1 minute readings all above a specified level
Monitoring Data Particles

Alarm Level
16

14 Alarm Event

12

10
PARTICLES

0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
MINUTES
 ISO 14644-2:2015 - Alert and Action Limits, How to choose the right strategy
STRATEGY 2
• Establish a trigger threshold value based on a high frequency of elevated readings. This method is
commonly referred as “x out of y”, where “x” is the number of events and “y” is the number of
minutes.
– For example, 3 out of the last 10 readings/minutes are above the specified alarm threshold
Monitoring Data Particles

Alarm Level
18
16
14
12
PARTICLES

10
8
6
4
2
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
MINUTES
 ISO 14644-2:2015 - Learn from FDA 483 Warning Letters
• Being compliant with any standard requires experience,
knowledge and a critical approach, enabling the harmonization
of regulation requirements with specific production
environments.
• Jumping on this task can be done well in advance by studying
the observations of FDA inspectors made to users in similar
situations.
• Several 483 warning letters have been redacted over the last
years, many of them strictly linked with monitoring plans' lack
of compliance. Some of them are summarized in Scott Sutton's
The Environmental Monitoring Program in a GMP
Environment.
 ISO 14644-2:2015 - Learn from FDA 483 Warning Letters
483 letter extract
• …Regarding the increased non-
routine surveillance monitoring
performed to further evaluate the
Building “123456” manufacturing
facility, there was no plan in place
specifying the locations to be tested,
method of sampling, and actions to
be taken when microbial
contamination was noted…
Comments
• This warning letter states the need of
having a monitoring plan in place, a
description of the location to be
tested and a specific sampling
method. Monitoring results are
considered insufficient if they don't
support and link to a clear and
approved plan.
 ISO 14644-2:2015 - Learn from FDA 483 Warning Letters
483 letter extract
• …the alert and action limits
established for the manufacturing
areas are not based on historical
data taken from the EM Program…
Comments
• This warning letter, dated 2001, requires the
cleanroom user to proactively and critically
review the sampling historical data, as it
must be referenced to correctly set up the
appropriate alert and action levels.
• This requirement was likely difficult to
accomplish in 2001, but is now easily
achievable using the appropriate software
platform (CFR 21 part 11 compliant), capable
with a particle counter's data storage.
 Conclusion
• The new changes described here will impact cleanroom classifications and
monitoring. Any company that needs to comply with this standard is required to
update their internal SOP in order to meet the new ISO 14644 requirements.
• ISO 14644-1/2:2015 are not only a new standards to be compliant with, but are also
beneficial tools to use in achieving mature cleanroom environmental control.
• The standard's main goal is to cultivate and promote a strong knowledge of
cleanroom performance inside every company.
• ISO 14644-1/2:2015 were published on December 15th, 2015. All users who want to
be compliant with this standard are required to take necessary action immediately.
Thank you for attending!
Daniele Pandolfi
Global Product Line Manager, Aerosol
dpandolfi@pmeasuring.com

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