Without measurement there is no control

Understanding ISO Standards:

ISO 14644-2:2015 Cleanroom Monitoring
Abstract
After the recent release of the ISO 14644-1:2015 Revision Summary and Understanding ISO 21501-4 application
notes, Particle Measuring Systems continues the ISO Standards interpretation series of documents with the
evaluation of the newly released ISO 14644-2:2015.
This document underlines the major changes between the previous version of ISO 14644-2 and the latest,
second edition, dated December, 12th 2015. A specific approach to the best practices to be adopted for an
efficient and compliant cleanroom monitoring process are discussed.

Introduction
In combination with the revision to Part 1. Classification of air cleanliness by particle concentration of ISO 14644-
1:2015, the ISO Technical Committee TC 209 has been working on the update to the basic airborne cleanliness
monitoring guidelines contained in ISO 14644-2.
The TC 209 community voted in favor of the revision to update and improve the standard specifically to:

• Simplify and clarify requirements and guidance tables that specify frequency of testing and monitoring
of cleanrooms used to demonstrate continued compliance with the cleanliness classification

• Refine how these intervals may be extended, provided that automated monitoring systems show the
cleanroom is under control

• Provide new guidance on aspects that should be considered when configuring a monitoring system for a
cleanroom
The new standard's introduction also lists the goals of the ISO 14644-2, when successfully applied:

• Emphasize the needs and advantages of a planned cleanroom contamination monitoring

• Provides the method for a correct particles contamination alarm and warning limits setting, based on a
careful evaluation of data trends

• Define the differences between a simple periodic cleanroom control and a more intensive/complex
monitoring strategy

• Enhance the installation and process knowledge, as to improve the risk assessment evaluation and a
faster reaction to any unexpected cleanroom performance

• Concretely reduce the operation cost by preventing production loss

[email protected] | +1 800 238 1801 Page 1 of 8

 Understanding ISO 14644-2:2015

Terminology
Before we get into the specific requirements, the understanding of ISO Standard terminology is essential. Below is
a list of the most important terms and their definitions:

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”.

Action Level Alert Level

User set level at which, when User set level, defined to provide early
exceeded, will require immediate warning of a drift from normal conditions.
intervention, root cause investigation This level should be used to prevent action
and corrective actions. level conditions.

Risk Assessment
The appropriate creation of risk assessment documentation is a basic requirement for implementing a monitoring
plan.
The new ISO 14644-2:2015 requires a well-developed risk assessment document as a tool to correctly understand
the process, the critical areas/locations, possible sources of contamination, and any element/event that may
compromise the cleanroom performance, product quality and, consequently, negatively affect the operation cost.

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. It is well integrated with good manufacturing practices
required by the Pharmaceutical Industry Standards.

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.
Other useful tools to be considered for reliable risk assessment development are HACCP,
FMEA/FMECA, PHA, FTA and HAZOP.
A responsible understanding of the production process and installation performance aids in
the prevention of unexpected out of specification conditions, and also assists in achieving
energy saving targets.

[email protected] | +1 800 238 1801 Page 2 of 8

 Understanding ISO 14644-2:2015

Monitoring Plan
Particle concentration control is mandatory for cleanroom classification (ISO 14644-1:2015) and monitoring (ISO
14644-2:2015). It provides clear evidence of cleanliness level and the capability of cleanrooms or clean zones to
accomplish required production performance.

CLEANROOM MONITORING CAN BE:

SEQUENTIAL
Sequential monitoring is normally performed by using multiplexing
manifold systems. This practice is well accepted in semiconductor
industries where small particle size (no greater than 1 micron) are
monitored.
The sequential monitoring method typically utilizes long transport tubes,
which is unacceptable in Pharmaceutical Manufacturing cleanrooms
where larger particles are considered and the risk of particle loss in
tubing isn’t negligible.
Read more about Particle Loss in Tubing. SEQUENTIAL

CONTINUOUS
The continuous method uses multiple particle counters, one per individual
location.
This monitoring approach provides a continuous flow of data over time, and
is normally active during the entire production phase.
The correct use of this control method allows for the immediate evaluation
of unexpected contamination events, thereby allowing for swift performance
CONTINUOUS
of corrective actions.
Pharmaceutical aseptic production is required to perform continuous particle
monitoring during the entire production time. This methodology provides trend
information useful for alarm and action level evaluation.

PERIODIC
The periodic method consists of performing particle monitoring at a scheduled
frequency (i.e., once per week), to demonstrate continuous compliance of the
cleanroom from one classification test to the next.
ISO 14644-2:2015 allows periodic monitoring to be performed, provided the user
clearly specifies the test frequency.

PERIODIC

[email protected] | +1 800 238 1801 Page 3 of 8

 Understanding ISO 14644-2:2015

The ISO 14644-2:2015 standard lists the main items to be taken into account when developing an airborne particle
monitoring system:

• Understand the contamination sources and their impact on the activity in the cleanroom

• Locate particle counter probes as close as possible to critical zones

• Potential adverse impact of the sampling system on the process or the process environment (e.g. possible
effects of the rate of the extraction of the sample volume in small volume environments)

• Airborne particle collection efficiency, suitability to monitor the selected particle size(s), and accessibility
for maintenance, calibration, and repair

• Air sample flow rate and volume

• Frequency and duration of the collection of each air sample (determined by the sampling rate)

• Sample probe configuration and orientation with respect to airflow (e.g. isokinetic or anisokinetic)

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.

• In case longer sample tubing is needed, the particle loss rate should be evaluated by measuring the
number of particles that remain trapped in the transportation tubes.

• Some method examples can be found in How to Conduct Particle Transport Tests.

WHAT’S THE RIGHT ISOKINETIC PROBE POSITION?

• The sampling probe (named Isokinetic Probe or ISP) provides the advantage of harmonizing the sampling
flow speed with the typical laminar flow patch speed (90 ft/min or 0.45 m/s), in respect to the ISO
requirements for the purpose of not creating any potential adverse impact of the sampling system
on the process.

• The position of these probes must be evaluated in the risk assessment and may never be
located right under the filtration system (unless specifically required).

• The position of the sampling probe must be representative of the cleanroom

performance and should be placed as close as possible to where the production occurs
and where risk to the product is highest. Also, it must be clearly determined and included
in monitoring SOPs to allow for reproducible sampling operations.

[email protected] | +1 800 238 1801 Page 4 of 8

 Understanding ISO 14644-2:2015

WHICH MONITORING METHOD SHOULD I USE?

ISO 14644-2:2015 does not specifically provide a link between the cleanrooms class cleanliness grade and the
recommended monitoring method, meaning that it is up to the user to choose the most appropriate one based
on their specific manufacturing requirements and risk assessment.
Life science industries may also consider other standards to correctly and reasonably set up their monitoring
method and frequency. For example, the 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 cleanroom cleanliness grade class
(see Table 1).

Table 1. Monitoring Frequencies for In Operation Routine Particulate Sampling

Classification In Operation (Dynamic) Routine Particulate Sampling

Grade A
For the full duration of operation
(filling operation)

Grade B Daily (working days)

Grade C Weekly

Grade D Not required

UDAF work stations in B Daily (working days)

UDAF work stations in C Weekly

UDAF work stations in D Monthly

UDAF in UNC areas Runtime re-qualification of UDAF is sufficient.

Table taken from Environmental Monitoring of Cleanrooms in Vaccine Manufacturing Facilities. WHO, 2012.

How to Set Action and Alert Limits

The determination of action and alert limits is extremely important, and must be supported by the risk
assessment as well as a consistent quantity of historical sampling data. In addition, they must be supported by a
consistent amount of data collected during previous monitoring controls.
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. For old plans especially, it should be questioned whether they are still applicable
and consistent with the cleanroom's actual performance, activities and needs.

[email protected] | +1 800 238 1801 Page 5 of 8

 Understanding ISO 14644-2:2015

The standard provides some important recommendations, as well as 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 particle concentration is more likely and
should be avoided by careful selection of alert and action levels. Frequent “nuisance” alarms
should be avoided as they can lead to alarms being ignored by users.
This concept is frequently ignored when setting alert or action alarms for large particle sizes in areas of high
cleanliness, and may result in extensive and useless investigation to determine the reason of alarms.
Good methods to deal with large particle sizes (i.e. 5 µm) in ISO 5 cleanrooms are described in paragraph B.3.4 of
the ISO 14644-2:2015 standards.
The following strategies consider the importance of evaluating an alert or alarm situation by taking into
consideration a series of events rather than a single spot value.

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.

[email protected] | +1 800 238 1801 Page 6 of 8

 Understanding ISO 14644-2:2015

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.

ISO 14644-2:2015 COMPLIANT – LEARN FROM FDA 483 AND TECHNICAL REPORTS
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.
Some issued letters may refer to basic concepts when read by cleanroom
experts, which remain a useful component when preparing a demonstration of
an efficient, compliant and defensible monitoring program.
The following 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.
…Regarding the increased non-routine surveillance monitoring
performed to further evaluate the Building 37 Flu 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…

[email protected] | +1 800 238 1801 Page 7 of 8

 Understanding ISO 14644-2:2015

The FDA emphasizes the need to develop a monitoring plan, based on risk
analysis. Compliance with the reference standards before implementation
must be verified. The following is an excerpt of the warning letter:
…The [redacted] method used for increased surveillance monitoring
of the environment has not been qualified…
One 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. Here is an excerpt of the letter:
…the alert and action limits established for the manufacturing areas LASAIR® III AEROSOL PARTICLE COUNTER:
are not based on historical data taken from the EM Program… CFR 21 PART 11 COMPLIANT

Conclusion
ISO 14644-2:2015 is not only a new standard to be compliant with, but is also a beneficial tool 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. This goal is accomplished by enforcing a continuing discussion and evaluation of collected sampling
data, development of user-set alert and action levels, and a proactive revision of monitoring plans and risk
assessments based on the ongoing developments of monitoring activities, rather than considering each single
monitoring session as isolated from the entire program.
ISO 14644-2:2015 was published on December 15th 2015, and all users who want to be compliant with this
standard are required to take necessary action immediately. For more information about the new ISO 14644-
2:2015 or other specific questions concerning cleanroom contamination control, contact your Particle Measuring
Systems local representative or navigate to our Contacts web page.

Author
Daniele Pandolfi

Daniele Pandolfi is the Global Product Line Manager, Aerosol in

Particle Measuring Systems’ Life Science Division. He has over
ten years’ experience in particle counter instrumentation and
cleanroom contamination control, built building close customer
relationships. He helps customers to solve their cGMP issues.

© 2016 Particle Measuring Systems. All rights reserved.

Lasair® is a registered trademark of Particle Measuring Systems.
Reproduction or translation of any part of this work without the permission of the copyright owner is unlawful.
Requests for permission or further information should be addressed to Particle Measuring Systems, Inc. at
1-800-238-1801.
App Note 246
01/2016

[email protected] | +1 800 238 1801 Page 8 of 8