Whitepaper

Changes to GMP force

cleanroom re-classifications
Tony Harrison, Senior Marketing Manager, Particle Counting & Characterization Business Unit

OVERVIEW
According to both USA cGMP 1 and EU GMP 2 cleanroom classification should be carried out according to ISO
14644-1. The 19994 version of this ISO standard has been re-written by a group of international subject matter
experts and ISO 14644-1:2015 6 was published on December 15, 2015. ISO 14644-1:2015 contains substantial
revisions impacting both the way the cleanrooms are classified and the performance requirements of the air
particle counting instruments used to carry out the classification. Cleanroom users claiming compliance to cGMP
and/or GMP will have to consider these new rules when next re-qualifying their cleanrooms, potentially purchasing
new air particle counters if their existing counters do not comply with the new performance rules and facing the
dilemma that the new re-qualification may indicate that their cleanroom is not operating at the grade they had
achieved using the old classification process. This paper explains the new rules.

Introduction
Although EU GMP and USA cGMP both define the airborne particle counts/m3 for each cleanliness class/grade,
they do not contain guidance on the cleanroom classification procedure. Instead they point the reader to ISO
14644-1 for the cleanroom classification methodology. As part of the routine periodic review process, the ISO
committee responsible for ISO 14644-1 decided that areas of the classification process required revision to improve
statistical accuracy. At the same time, the ISO committee decided to add a reference in the normative Annex on
airborne particle counters to
ISO 21501-45, which provides a calibration procedure and verification method for particle counters, intended to
minimize the inaccuracy in the measurement result by a counter, as well as the differences in the measured results
from different particle counters. The stringent performance rules in ISO 21501-4 mean that many older air particle
counters are no-longer suitable for cleanroom classification/re-qualification according to ISO 14644-1.

Cleanliness Classes/Grades
There are fundamental differences between the USA cGMP and the EU GMP documents for cleanroom classes/
grades. The foremost difference is that the EU GMP requires the cleanroom user to classify and subsequently
monitor their cleanrooms for airborne particles 0.5microns in size and also 5microns in size, whereas the
USA cGMP only requires the user to classify and subsequently monitor their cleanrooms for airborne particles
0.5microns only.

As can also be seen from the tables in Fig. 1 and 2, cGMP does not define airborne cleanliness levels for each
cleanroom Class when the cleanroom is at rest, whereas GMP differentiates between these two states.
The third major difference between the two documents is that cGMP has an additional Class between Class
100 (equivalent to GMP Grade A) and Class 10,000 (equivalent to GMP Grade B in operation) and GMP has an
additional Grade D, defined as having 3,520,000 particles 0.5microns at rest.

TABLE 1 - Air Classifications

Clean Area
Classification
3
(0.5 mm particles/ft )

ISO
Designation

100
1000
10,000
100,000

5
6
7
8

0.5 mm
3

particles/m

Microbiological
Active Air Action
Levels (cfu/m )

Microbiological Settling
Plates Action Levels
(diam. 90mm; cfu/4 hours)

1
7
10
100

1
3
5
50

3,520
35,200
352,000
3,520,000

Figure 1. Cleanroom Classes as defined in USA cGMP

Grade
A
B
C
D

Maximum permitted number of particles per m33 equal to or greater

than the tabulated size
At rest
In Operation
0.5 mm
5.0 mm
0.5 mm
5.0 mm
3 520
3 520
352 000
3 520 000

20
29
2 900
29 000

3 520
352 000
3 520 000
Not defined

20
2 900
29 000
Not defined

Figure 2. Cleanroom Grades as defined in EU GMP

The Changes to ISO 14644-1

The fundamental changes in the ISO 14644-1:2015 document impacting on the GMP community are:
1. Change to the method for calculating number of sample locations for classification/re-qualification
2. Elimination of the 95% Upper Confidence Limit (UCL) calculation
3. Reference to ISO 21501-4 in the normative* Annex A where the requirements for air particle counters are
defined (*normative = must comply with this section)
4. Removal of the maximum allowable concentration of airborne particles/m3 5microns for ISO Class 5 from the
Cleanliness Classes table (ISO Class 5 at 5microns is equivalent to GMP Grade B at rest)

The new method for calculating the number of sample locations now relies on a look-up table where the number of
locations per cleanroom size is defined as opposed to the current method that calculates the number of locations
based on the square root of the cleanroom floor area. In order to achieve a pragmatic level of statistical accuracy,
the number of sample locations required for classification/re-qualification has been increased, i.e. cleanroom users
will have to increase the number of sample locations for cleanroom classification/re-qualification. As per the ISO
14644-1:1999 document, ISO 14644-1:2015 states that the sample locations should be spaced out evenly across the
cleanroom on an equally-spaced grid.

The requirement for the new sample locations could lead to confusion in particle counter users who are used
to following the same classification/re-qualification routine, especially for those with multiple cleanrooms and
multiple air particle counter users. Cleanroom owners could attempt to mitigate the chances of resultant data
errors/miss sampling by choosing to use air particle counters that can be programmed with the new cleanroom
sampling program so that the user is prompted to take the correct number of samples using the correct particle
counter configuration in each cleanroom Grade. The same opportunity for confusion and potential resultant
mistakes would apply to any form of manual data transfer from paper print outs from the particle counters to
Excel* or a laboratory information management system (LIMS). Cleanroom owners could choose to mitigate the
risk of data transfer errors by choosing particle counters that can transfer the particle count data via wireless
Ethernet in Excel readable format straight to their secure data server or LIMS systems (visit http://www.particle.
com/met-one-air-particle-counters/portable/met-one-3400-simply-paperless for more information on particle
counters with these capabilities)

By increasing statistical accuracy through a larger number of sampling locations, the ISO 14644-1:2015 document
also eliminates the need for the UCL calculation which was previously required for cleanrooms requiring between
2 and 9 sample locations. Provided all sample locations are within the Class/Grade requirements, there is nolonger any need to perform the UCL calculation.

ISO 21501-4 lays down the calibration procedures to ensure that air particle counter sampling measurement
inaccuracies are reduced. It also includes a final confirmation of air particle counter performance, the Counting
Efficiency test. This compares the performance of the particle counter under test against an instrument with
a higher resolution of around 0.3microns. Both the counter under test and the higher resolution instrument
are challenged with an air sample containing certified particles at the smallest calibration size for the counter
under test, typically 0.5microns for GMP applications. As the certified particles will have an expected Normal (or
Gaussian) size distribution profile, 50% of the certified particles will be 0.5microns and 50% will be 0.5microns,
hence the counter under test must count and report that 50% of the airborne particles are 0.5microns, whereas
the instrument with the higher resolution should report 100% of the airborne particles as being 0.3microns.
Hence the higher resolution counter will count the entire particle population, allowing the ability of the counter
under test to accurately report that 50% of the certified particles are 0.5microns (see Fig. 3).

Figure 3. The ISO 21501-4 Counting Efficiency Test

In another move to improve the statistical accuracy of the classification/re-qualification, the international subject
matter experts tasked with re-writing the standard decided to remove from the Cleanliness Classes table numbers
of particles that were too small to be considered statistically sound. Controversially, this was extended to include
the 29 particles/m3 for ISO Class 5 for particles 5microns, potentially undermining the EU GMP requirement to
the measure these particles in Grade A and Grade B environments.

Implications for the GMP Cleanroom Community

Thanks to the undated reference to ISO 14644-1 in the GMP and cGMP documents, to the new ISO 14644-1:2015
will impact on every cleanroom user in the GMP community.
All GMP cleanroom users will have to make changes to their cleanroom classification/re-qualification SOPs to
incorporate the larger number of sampling points required in the new version of the ISO standard. In some cases a
risk assessment may be required to retrospectively investigate the potential impact on product quality if the new
classification/re-qualification changes the cleanroom Class or Grade. In some cases where this is the case, the
cleanroom owner may have to consider making changes to their cleanroom and air handling systems to bring the
cleanroom back into the required Class/Grade.
All GMP cleanroom users will have to ensure that the particle counters used to classify and monitor their cleanrooms
are compliant with the requirements of ISO 21501-4, including those counters forming a part of a continuous
monitoring system.

Conclusion
The changes to ISO 14644-1 represent possibly the biggest change to GMP cleanrooms since the requirement
to sample 1m3 at each and every sample location when classifying/re-qualifying Grade A environments was
introduced to GMP Annex 1 in 2003. The GMP cleanroom community would be wise to start planning for these
changes now rather than waiting till they are inspected to the ISO 14644-1:2015 doument.. (visit http://www.
particle.com/met-one-air-particle-counters/portable/met-one-3400-simply-paperless for more information on
particle counters designed to support the new ISO 14644-1 and ISO 21501-4 requirements)

*Excel is a registered trademark of Microsoft Corporation in the United States and/or other countries.

References
1. Food and Drug Administration. Guidance for industry. Sterile drug products produced by aseptic processing
current good manufacturing practice, 2004. U.S. Department of Health and Human Services Food and Drug
Administration Center for Drug Evaluation and Research (CDER) Center for Biologics Evaluation and Research
(CBER) Office of Regulatory affairs (ORA) Division of Drug Information, HFD-240 Center for Drug Evaluation and
Research Food and Drug Administration 5600 Fishers Lane Rockville, MD 20857 USA
2. European Commission. EudraLex. The Rules Governing Medicinal Products in the European Union. Volume 4.
EU Guidelines to Good Manufacturing Practice. Medicinal products for human and veterinary use, Annex 1:
Manufacture of Sterile Medicinal Products, 14th February 2008. European Commission Enterprise and Industry
Directorate-General, B-1049 Bruxelles / Europese Commissie, B-1049 Brussel Belgium.
3. European Commission. Ad Hoc GMP Inspections Services Group. EC Guide to Good Manufacturing Practice.
Revision to Annex 1. Manufacture of Sterile Medicinal Products, 30th May 2003
4. ISO 14644-1:1999(E) Cleanrooms and associated controlled environments Part 1: Classification of air cleanliness
1st May 1999. International Organization for Standardization Case Postale 56 CH-1211 Genve 20 Switzerland
5. ISO 21501-4 Determination of particle size distribution Single particle light interaction methods Part 4: Light
scattering airborne particle counter for clean spaces 2007. International Organization for Standardization Case
Postale 56 CH-1211 Genve 20 Switzerland
6. ISO 14644-1:2015 Cleanrooms and associated controlled environments Part 1: Classification of air cleanliness
by particle concentration. International Organization for Standardization Case Postale 56 CH-1211 Genve 20
Switzerland

Biography
Tony held the Convenorship of the ISO Working Group revising ISO 14698-1 & -2 for microbial control in cleanrooms
and is the UK subject matter expert to the ISO Working Group currently revising ISO 14644-1 & -2 for cleanroom
classification at the heart of the aseptic manufacturing chapters of both the European GMP and the USA cGMP
documents.
Tony holds a Bachelors Degree in Electrical & Electronic Engineering and is employed by Beckman Coulter Life
Sciences as a Senior Marketing Manager.
Experienced in water system TOC, conductivity and ozone analysis and cleanroom monitoring systems as well as
particle characterisation, Tony has spent the last fifteen years in applied metrology for the pharmaceutical and
healthcare manufacturing industries. Prior to that, he worked for companies providing process control automation
solutions for manufacturing industries.
Tony was joint-editor on the ISPE Guide to Ozone Sanitization of Pharmaceutical Water Systems and was also chief
editor of the PHSS Best Practice Guide for Cleanroom Monitoring.
Tony is a well-known international speaker and has provided educational seminars on TOC, liquid particle counting,
ozone sanitization for water systems and cleanroom monitoring in UK, France, Italy, India, Malaysia, China, USA,
Scandinavia, Ireland, Hungary, Switzerland, Indonesia, Belgium, Greece, Switzerland, Turkey, Egypt, Denmark and
most recently Poland and Italy.

Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries.
For Beckman Coulters worldwide office locations and phone numbers, please visit Contact Us at beckmancoulter.com
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