Revision of ISO 14644-1:1999 A progress report and

explanation of some of the key issues and principles

June 2012
Gordon J Farquharson
Convenor ISO TC209 WG1, Critical Systems Ltd, 4 Greencroft, Guildford, Surrey, GU1 2SY

Corresponding author: Gordon J Farquharson, Critical Systems Ltd, Guildford, Surrey, GU1
2SY, UK; email: gj.farquharson@gmail.com

Introduction
I have taken the unusual step of using some pages of this journal to share some very
important issues relating to the revision of the fundamental cleanroom standard ISO 146441:19991 (ISO: International Standards Organisation). I decided to do this because the
specifications of air cleanliness by particles form a fundamental part of our good
manufacturing practices (GMPs) for sterile products in particular, and many other active
pharmaceutical ingredients (APIs) and dosage forms. The pharmaceutical and life sciences
industry uses cleanroom technology in many parts of its manufacturing and research
operations, and, in these applications, the control of air cleanliness has a direct link to
regulatory compliance and achievement of specified product quality.
I have not explored the parallel process of revision of ISO 14644-2:20002 since,
whilst an important standard, the issues at large are not as fundamental to the cleanroom
community.
The broader cleanroom community, beyond the pharmaceutical and life-sciences
industry, is also directly influenced by the evolution and development ISO 14644-1:1999,
whick is indeed a very important standard. The process of revision involves input by
technical experts within an ISO committee environment. These experts should bring to the
table current scientific and technical knowledge on the subject, and also table the opinions
and requirements developed through national mirror organisations.
This is not the first time that this journal has addressed the subject. This journal
published a paper in January 2011 (Hartvig, et al.)3, which was prepared to explain the basis
of the sampling statistics in the ISO DIS 14644-1:20104 (DIS: Draft International Standard).

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This report builds on the principles developed in that paper, and sets out to explain the areas
of concern, technical argument, and in particular where the standards community reported
difficulties with the relationship of this DIS to the pharmaceutical GMPs.

Publication of ISO DIS 14644-1:2010 for review and national vote

Close followers of the ISO standardisation process for cleanrooms under ISO TC209 (TC:
Technical Committee) will recall that a draft international standard, ISO DIS 14644-1 was
published at the end of 2010 for the statutory 6-month DIS review, comment and vote. As has
been the case with the complete family of ISO TC209 cleanroom standards, the technical
enquiry and vote was undertaken in parallel within the CEN standards community (CEN:
Committee for European Normalisation). A convention, called the Vienna Convention, put in
place to prevent unnecessary parallel work being carried out in CEN and ISO environments,
has been applied to this work.
The DIS enquiry and vote is a very important stage in the standardisation process
because it is the first time that the draft documents are opened beyond ISO TC209 for broader
public and industry scrutiny. Equivalent English, French and German language versions are
published by national standards bodies, and formal comments are sought. The comments are
then submitted back to the ISO central secretariat, together with a national vote. In the case of
the recent cleanroom standards process, the national voting was in favour of approval of both
ISO DIS 14644-1 and ISO DIS 14644-2 subject to comments being addressed. The ISO and
CEN rules require that all comments are considered and adjudicated upon and reported for
the record.

Extent of comments
I would now like to move on to explore the scale and nature of the comments made during
the DIS enquiry process between December 2010 and May 2011. I am going to focus on ISO
DIS 4644-1:2010 because this is the more important of the two standards, and one which
received the majority of substantive comments. There were more than 100 substantive
technical comments, and many more of an editorial nature. Every one of the comments has to
be assessed and reported on.
ISO TC209 WG1 (WG: Working Group) met on the 8th and 9th October 2011 to
consider the comments and the main issues raised by industry (WG committee of experts
nominated by national standards bodies to develop the content of particular standards). The
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nature of the comments, and the likely changes to the DIS were so significant that a second
DIS enquiry and vote will be required. This means a delay to the revision project. However,
the topic of cleanroom classification is so significant that the concerns of industry have to be
addressed.
The majority of the comments came from the pharmaceutical and life sciences industry, but
there were also some very important comments made by semiconductor and display screen
manufacturing industry. One of the most interesting of these was a serious concern about how
the standard should deal with super-large cleanrooms. To help experts understand scale, a
Chinese presentation was made that illustrated single cleanroom facilities of up to 40,000m,
with working zones of up to 10,000m. These are clearly outside the range of expectation
envisaged during the evolution of cleanroom standards from the earliest US Federal Standard
209, up to the current ISO 14644-1:1999, and also the ISO DIS 14644-1:2010. I will address
this issue later, when we consider the number of sample locations required for classifying a
cleanroom or clean-zone.
Sadly, I have to report that many of the national comments on the draft standard
challenged or contradicted some of the important principles developed by their own experts
during the 2-year evolution of the new draft. Whilst this is extremely disappointing I took the
view, as convenor, that it was more important to work through the issues to ensure the
broader acceptance of a revised standard in the future. When challenged, these experts
explained that the DIS had been opened to broader scrutiny than had originally been possible
in their national mirror committees.
The main areas of concern identified in the vote and comments were as follows.

The relevance of the particle concentration values in the classification table.

The exclusion of particles 5 microns from the classification table for ISO Class 5.

The increased number of sample points required for classification of a cleanroom or

clean-zone compared to the current 1999 version.

The locating of the sample points within the cleanroom or clean-zone (randomisation
issues).

Dealing with risk-based locations selected to specifically certify the cleanliness of

critical locations within a cleanroom or clean-zone.

The comments and issues in more detail

The role and significance of the classification table
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The subject I want to look at in more detail is Table 1 in both the current standard (Figure 1)
and the proposals in the 2010 DIS (Figure 2). The first most important change between the
existing standard and the DIS is that Table 1 becomes the foundation and basis of
classification. The advantage of doing this compared with the use of formula (as in the 1999
version), is that the table can be used more effectively to constrain and guide readers and
users into choosing appropriate particle sizes and cleanliness levels. Historians of cleanroom
standardisation will remember that this was the basis of US Federal Standard 209E5 and its
US predecessors.

Figure 1. Informative Table 1 from ISO 14644-1:19991

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Table 1 Classification table for particulate cleanliness classes

ISO Class 1

Maximum allowable concentrations (particles per m3) for

particles equal to and greater than the considered sizes shown
below a
0,1 m 0,2 m 0,3 m 0,5 m 1 m
5 m
d
d
d
d
e
10 b

ISO Class 2

100

24 b

10 b

ISO Class 3

1 000

237

102

35 b

ISO Class 4
ISO Class 5

10 000
100
000
1 000
000

2 370
23 700

1 020
10 200

352
3 520

83 b
832

237 000

102
000

35 200

8 320

293

ISO Class 7
ISO Class 8

83 200
832 000

2 930
29 300

ISO Class 9

352 000
3 520
000
35 200
000

8 320
000

293 000

ISO Classification
number (N)

ISO Class 6

Figure 2. Normative classification Table 1 from ISO DIS 14644-1:20104.

Figure 3. Equivalent table found in US Federal Standard 209E5.

The following became apparent from review of the comments made on Figure 2
above. The issue equally applies to Figures 1, 2 and 3. Many readers were of the opinion that
for a given ISO Class 19, the tabulated particle concentrations at different particle sizes
actually represents typical particle distributions found in the cleanroom environment. This is
not the case. A number of experts, including workers such as Whyte, Eaton, Ljungqvist and
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Reinmuller have all carried out particle concentration measurement and assessment work that
suggests very clearly endorses this position. During the ISO TC209 WG1 meeting in October
2011, the experts took account of these facts, and even considered proposing a new table that
might better represent typical particle distribution at different sizes in cleanrooms. After
much deliberation, it was decided that this was a step too far because it would mean a
fundamental change to the current understanding of the ISO Classes 19. Following further
discussion, it became clear that this only presents a problem when you try to use more than
one particle size to classify a level of cleanliness. Within the existing 1999 standard, the last
paragraph of clause 3.3 suggests it is legitimate to use more than one particle size, provided
there is a reasonable separation in the sizes considered. One possible simple solution to this
would be to simply remove this clause. At this juncture, it is important to remember that ISO
14644-1 is a standard specifically constructed for the purpose of carrying out a cleanroom
classification. This is a formal process undertaken during the commissioning of a new
cleanroom asset, and then periodically thereafter to prove continued compliance with the
class. In addition to a formal classification, most practitioners, either based on good business
practice, or as mandated by a regulatory authority, additionally monitor air cleanliness by
particles is in real-time. Under these circumstances, selection of more than one particle size
might be more appropriate because one should really be looking at deviation from a state of
demonstrable control, rather than compliance with a relatively arbitrary value.

The 5 micron particle problem

The removal of the 5 micron particle concentration in ISO 5 compared to the 1999 version
was a major concern for a number of reviewers (see Figure 2 above, and compare with
Figure 1). In the 1999 version, the figure, of course, was the well-known 29 particles per
cubic metre (see Figure 1). So why did experts feel the need to remove this 5 micron particle
number concentration? The clear advice from the experts was that this low concentration of
large particles is unreliable to count using our airborne particle counters, and, therefore, is
considered an inappropriate size of particles to evaluate the ISO Class 5. Once again, there is
a clear precedent for this in that this was exactly the situation in the discontinued US Federal
Standard 209 E (see Figure 3). It is quite difficult to find a clear reason why the 29 particles
limit was retained in 1999, but it is most likely to have occurred because ISO 14644-1:1999
included a significant European input. It is really important to realise that once the new
standard is agreed as a classification based on Table 1, and the table includes no figure in the

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5 micron column for ISO Class 5, then this means that you cannot use this particle size for
classification in accordance with ISO 14644-1.
This leaves us with an interesting problem about how to support the regulatory
requirements set out in Annex 1 of the EU GMP. In the view of particle counting experts on
WG1, the concentration of 5 micron particles in grades A and B of Annex 1 are just as
inappropriate as 29 particle concentration in the ISO standard when considering
classification. Monitoring may be a different matter, not considered in any version of ISO
14644-1. This is becoming an even more sensitive issue due to the recent replication of the
EU GMP Annex 1 requirements in the PIC/S GMP, the WHO GMP, and most recently the
new Chinese GMP regulations. It is because of this that many comments received on ISO
DIS 14644-1:2010 requested that the 29 particle limit be replaced. Whilst this matter is not
yet concluded, the expert opinion is still that Table 1 in the cleanroom standard should not
include particle concentration sizes that are inappropriate. The experts are currently working
on forms of words, to be included in the standard, which would allow pharmaceutical
industry regulatory authorities to provide their own guidance outside the boundaries of the
standard.
The nature of the guidance being considered very seriously is to recommend complete
separation of the activities of cleanroom or clean-zone classification from real-time
monitoring. This is already clearly stated in Annex 1. Careful reading of the text in Annex 1
will indicate to the reader that the regulatory authorities are most interested in the operational
state. The classification of cleanrooms or clean-zones in the at-rest state is more about
defining a benchmark of effective performance of the environmental control system to give
confidence that there is no external contamination contribution within the controlled
environment. The real-time monitoring in operation, on the other hand, gives you much more
important information about the sources of contamination immediately around the process in
operation. The greatest concerns relate to the contamination sources from human beings, and
failed technical systems. Under these circumstances, it would be much easier to consider 5
micron particle assessments for real-time monitoring alone. One of the greatest advantages of
this approach would be that it would allow a better fit with the scientific approach to the
revision of ISO 14644-1, and would remove some of the concerns being expressed about
increased testing time due to the combination of a tight 5 micron particle limit and, in some
cases, an increased number of sample locations proposed.

Number of sample locations

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The next important subject we have to consider is the basis of the new table of sample
locations. The experts believe that a look-up table is a more appropriate approach because it
allows the cleanroom or clean-zone to be divided into fundamental unit areas that have some
tangible logic. Therefore, for very small clean-zones, fundamental unit areas of one square
metre were considered appropriate, and for larger cleanrooms and clean-zones fundamental
unit areas of 4m were considered to be suitable. The DIS feedback clearly indicated that
there was confusion between fundamental unit areas and sub-divided regions. The experts
are working on improvement to the text to resolve this problem.
It is a basic assumption within the DIS standard, and, indeed, a similar principle exists
in the current standard, that within any single fundamental unit area the cleanliness level is
assumed to be even throughout the area, and that a sample taken anywhere in a fundamental
unit area will be representative of the whole area.
The DIS feedback clearly demonstrated that the majority of the cleanroom community
accepted that an increase in the number of sample locations to give at least 95% confidence
that at least 90% of all locations do not exceed the class limits was an adequate level of
assurance. It also allows the 95% upper confidence limit evaluation required for between 2
and 9 locations in ISO 14644-1:1999 to be discarded. This simplifies the classification
process, one of the demands made by the user community at the outset of this revision.
We now have to consider the issue of super-large cleanrooms. This was a problem
brought to the table by Chinese experts, and the main concern was that the proposed new
table of sample locations would lead to unnecessarily large numbers of sample locations for
very large rooms. The Chinese experts reported that they felt comfortable with a combination
of sample locations based on square root of the area of clean-zone, plus specific risk-based
process-specific test locations. The problem is best illustrated by considering a single
cleanroom area of 10,000m. In such a situation, if we use the square root rule in ISO
14644:1999, then we would require only 100 locations to carry out full classification. If you
take the sample location requirement identified in ISO DIS 14644-1:2010, then you will
require 520 locations because the draft stated that you have to divide the cleanroom into
500m parts, and apply the sample location table to each part. Once again, this is still under
consideration by the experts, but it is likely that the table of sample locations will be used for
up to 900m, and then the square root rule would be applied for rooms or clean-zones greater
than 900m. The logic of this can be seen on the graph below, where 900m is the point where
the confidence level and the square root rule lines cross (see Figure 4).

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Square root rule

ISO 14644-1:1999
Applies above 900m2
Transition point at
900m2
95/90 rule
ISO DIS 14644-1:2010
Applies up to 900m2

Figure 4. Sample point number comparison.

From a statistical point of view, the confidence level would theoretically be much
higher for cleanrooms greater than 900m than for those less than 900m. The WG1 experts
believe this is a reasonable pragmatic approach to give a reasonable number of locations. The
other aspect of super-large cleanrooms that requires some additional consideration is those
which have an extremely high ceiling. The experts believe the new DIS should contain an
element that explains that you may need to define several test planes within the cleanroom,
and carry out classification at multiple locations in each test plane.

Random selection of sample locations

This was the next bridge confronting the expert committee review. During the course of the
development of the DIS, the experts had agonised over how to deal with the random selection
of previously established fundamental unit areas. Initially, the concept developed involved
dividing the cleanroom or clean-zone into equal areas based on the number of fundamental
unit areas, and then randomly selecting from these the number of areas required to be
sampled from the look-up table providing a number of sample locations. So, if we had 100m
of clean-zone, this would be divided into 25 fundamental unit areas, and from the look-up
table, we would be required to sample from 16 of them. The 16 would be selected randomly.
There is, of course, the possibility that the 16 could be grouped together, rather than be
evenly distributed (Location Selection Option 1). To get over this, the DIS adopted a semiPage 9 of 12

random approach that tried to ensure a more even distribution of sample locations (Location
Selection Option 2). This received quite a lot of negative response during the DIS enquiry.
The experts have further discussed these two options at length, and now the majority believe
that the original plan should prevail. It is most likely that Location Selection Option 1 will be
worked on in more detail in the next few months.
Next we need to consider how to undertake the random selection. This becomes a
great deal easier once we decide to choose Location Selection Option 1. A standard number
generator without replacement can be easily found on the Internet for this kind of activity.

Terminology used
There is also some confusion about terminology used in the context of subdividing a
cleanroom or clean-zone, with terms used such as sub-zone, region, area, sector and so on.
These matters will be resolved by the experts, and will be coordinated with the other
standards in the ISO TC209 family. The likely outcome of this, subject to further review and
discussion, will be the following:

cleanroom;

clean-zone;

fundamental unit area;

number of sample points;

sample point location.

Conclusion and preview of the next draft ISO 14644-1

I want to try to summarise this complex story, and suggest that the following scope and
principles are the most likely scenario for the second DIS 14644-1 to be published for
comment and national vote sometime in 2012.
1. The classification will be by means of a table quite close to that in Figure 2, and will
exclude the 5 micron particle number concentration for ISO 5. It should be noted that
the table does not address the real-time monitoring, or special requirements for testing
the presence of 5 micron particles as required by some regulatory authorities.
2. The sample size calculation will be based on exactly the same principle as in the
current ISO 14644-1:1999. That is, you must sample sufficient volume that you would
count 20 particles at the class limit for the largest considered particle size for the ISO

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cleanliness class in question. This is the requirement within the existing standard and
endorsed in the DIS vote, and was not questioned in the vote response.
3. There will be a look-up table that provides information for a cleanroom on the number
of fundamental unit areas, and the number of these that needs to be selected for
sampling. Figure 5 illustrates how this might look, and includes some sample values.
Beneath the table will be a description on how to divide up the cleanroom or cleanzone, and how to use a random number generation process for selecting the ones to be
tested. The standard will specify a procedure something like the following.

Identify the number of fundamental unit areas from the table.

Divide the total area of cleanroom or clean-zone into equal sized fundamental
unit areas and identify them uniquely.

Identify from the table the number of fundamental unit areas that need to be
tested.

Use a random number generator to define the ones to be tested.

Locate the sample point anywhere within the defined fundamental unit areas
for this test. (Since there is a basic assumption that each fundamental unit area
is homogeneous in terms of cleanliness, then the absolute location of the
sample point within the randomly selected fundamental unit areas can be
determined by the tester.)

Should additional risk-based sample points be required, then these are

additional to those selected on a random basis, except when a risk-based
sample point happens to be in one of the randomly selected fundamental unit
areas.

Cleanroom or clean-

Fundamental

Number of available

Minimum number of

zone available area m2

unit area used

fundamental unit areas

fundamental unit areas required

m2

to be tested

36

52

13

10

104

26

16

116

29

18

900

225

29

>900

Use the square root of the area and the practice defined in ISO 14644-1:1999. (Note: the details of
how this transition works are still to be developed by the experts of ISO TC209 WG1.)

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Figure 5. Example sampling plan look-up table.

4. Take the minimum air sample at each point, and count the concentration of particles.
This concentration would be normalised to particles per cubic metre. Should more
than one sample be taken at any sample point, then it is acceptable to average the
count prior to normalising the value.
5. The data collected shall then be evaluated on a point-by-point basis, and all points
must comply with the class limit for a specific classification class.

References
1. International Standards Organisation. ISO 14644-1:1999 Cleanrooms and associated
controlled environments Part 1: Classification of air cleanliness. Geneva,
Switzerland: ISO; 1999.
2. International Standards Organisation. ISO 14644-1:2000 Cleanrooms and associated
controlled environments Part 2: Specifications for testing and monitoring to prove
continued compliance with ISO 14644-1. Geneva, Switzerland: ISO; 2000.
3. Hartvig NV, Farquharson G, Mielke R, Foster M. Sampling plan for cleanroom
classification with respect to airborne particles. EJPPS 2011; 16(1):512.
4. International Standards Organisation. ISO DIS 14644-1:2010 Cleanrooms and
associated controlled environments Part 1: Air cleanliness classification by particle
concentration (ACP). Geneva, Switzerland: ISO; December 2010. (Published by ISO
for the DIS enquiry and national vote.)
5. US-FED-STD-209E Federal Standard Airborne particle cleanliness classes in
cleanrooms and clean-zones. Published by US GSA June 16, 1988.

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