Professional Documents
Culture Documents
Revision To Parts 1 and 2: 2015 Tim Sandle
Revision To Parts 1 and 2: 2015 Tim Sandle
Tim Sandle
Welcome
Pharmig is a non-profit making professional organisation, established
in 1991, that represents the interests of individuals who work in, have
responsibility for, or work alongside microbiology within
pharmaceutical, healthcare, cosmetics & NHS Industries.
It provides a focus for continuing professional development and serves
as a unique network for the exchange of microbiological information
through training courses, conferences, publications and its website
forum.
Organising meetings, training courses, conferences and producing publications
that provide topical information and views on microbiologically related topics
Advancing the science of microbiology and its practical application
Influencing the development of regulations and guidelines
surrounding microbiology
Acting as a confidential forum for the dissemination of information
concerning all aspects of microbiology
Presenter
Dr. Tim Sandle
Pharmig committee
member
Pharmaceutical
Microbiology website:
http://www.pharmamic
roresources.com/
ISO 14644
ISO 14644
International cleanroom standard
Part 1 in 1999
12 part standard
Electronics, healthcare, biotechnology, pharmaceuticals
Replaced FS 209E in 2001
Accepted by EU GMP in 2003 for classification but not
monitoring
FDA aseptic filling guide in 2004.
ISO 14644 – parts #1
ISO 14644-1:2015 - Part 1: Classification of air
cleanliness
ISO 14644-2:2015 - Part 2: Specifications for testing
and monitoring to prove continued compliance with
ISO 14644
ISO 14644-3:2005 - Part 3: Test methods
ISO 14644-4:2001 - Part 4: Design, construction and
start-up
ISO 14644-5:2004 - Part 5: Operations
ISO 14644 – parts #2
ISO 14644-6: 2004 - Vocabulary
ISO 14644-7:2004 - Part 7: Separative devices (clean air hoods, gloveboxes,
isolators and mini-environments)
ISO 14644-8:2013 - Part 8: Classification of air cleanliness by chemical
concentration (ACC)
ISO 14644-9:2012 - Part 9: Classification of surface cleanliness by particle
concentration
ISO 14644-10:2013 - Part 10: Classification of surface cleanliness by chemical
concentration
No part 11 in draft
ISO 14644-12:draft - Part 12: Classification of air cleanliness by nanoscale
particle concentration
ISO 14644-13:draft - Part 13: Cleaning of surfaces to achieve defined levels of
cleanliness in terms of particle and chemical classifications
ISO 14644-14:draft - Part 14: Assessment of suitability for use of equipment by
airborne particle concentration
Cleanrooms
• Cleanrooms
– Design
• Air
– Filtration (HEPA)
– Pressure differentials
– Air changes
– Clean-up times
– Personnel
– Gowning
– Behaviours
In Operation:
NL = A
NL is the minimum number of sampling locations (rounded up to a whole
number).
A is the area of the cleanroom or clean zone in square metres (m2) for
which the square root is taken.
• Taking the surface of the room in square metres, assessing the square
root and using the obtained number (rounded up) to give the number
of locations, to be positioned equidistantly.
Change A– particle locations #2
Change A– particle locations #3
• The new method is based on hypergeometric distribution.
Opposite to binomial distribution.
Samples are drawn randomly without replacement from a finite
population.
Each location can be treated independently with a 95% level of
confidence that at least 90% of the cleanroom will comply with
the maximum particle limit for the intended class.
Confidence an be increased if desired.
No calculations are required to determine the number of
locations - there is a ‘look-up table’ (Table 1 - the only reference
for all sizes of particle from ISO 1 to ISO 9).
Look up tables #1
Where a room area is not
listed in the look-up
table, the next largest
size is selected.
This method has
generally led to an
increase in particle count
locations.
Look up tables #2
Cleanroom Room size 1999 version Revised no.
location of locations
numbers
A 200 m2 15 23
B 36 m2 6 9
C 8 m2 3 4
Change B – position of counters #1
• Once the number of locations has been selected, the room
is divided up into equal sectors and a particle counter
placed in each sector.
Previous standard – counter placed in approximate centre.
New standard - where the counter is placed within each sector
is determined by the user.
The standard allows counters always to be placed at the same
point within the sector; randomly placed within the sector; or
evenly distributed; or by risk.
Reason: counts no longer assumed to be homogenous within a
sector.
Addition locations can be added at the discretion of the
facility.
Change B – position of counters #2
To align with GMP, the location should be orientated
to the point of greatest risk e.g. close to fixed
equipment. The standard recommends that the
following is accounted for:
Room layout;
Equipment layout;
Airflow patterns;
Position of air supply and return vents;
Air-change rates;
Consideration should be given to any unintended bias in the
sampling process.
Change C – sample volumes #1
Volume of air to be sampled at For example, Grade C
each location, the volume of air
must be sufficient to detect at Volume to be sampled
least 20 particles for the largest
particle size limit. = 20 x 1000 = 0.69 litres
The operative figure is ≥5.0 29,000
microns
Therefore, a minimum of 1 litre
Volume to be sampled (Vs) = would need to be taken at each
location.
However, ISO 14644 states that
[20 x 1000 (constant)] the volume needs to be at least 2
Class limit particles (largest size) litres, sampled over a one
minute period.
Therefore, a minimum of 2 litres
would need to be taken at each
location.
Change C – sample volumes #2
For example, Grade B
Volume to be sampled =
20 x 1000 = 6.9 litres
2,900
Therefore, a minimum of 7 litres would need to be
taken at each location.
Grade A – more complicated…
Change D – class limits #1
The Grade A issue
EU GMP Grade A does not equal ISO class 5, because of
the different 5.0 µm limits
29 count limits for ISO 14644 class 5
20 count limit for EU GMP Grade A.
Where intermediate classes are required the standard no
longer permits increments of 0.1. So, to meet EU GMP, an
ISO class of 4.5 would need to be selected in theory.
Change D – class limits #2
De-emphasis on the 5 μm ISO Class 5 limit:
Sampling and statistical limitations for particles in low concentrations make classification
inappropriate; and
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.
But Annex 1 of EU GMP requires 5.0 µm particles to be
assessed
Options:
Just classify Grade A for 0.5 µm and use 0.5 µm / 5.0 µm for operations,
Or continue with 20 or 29 as a limit as an additional option for 5.0 µm.
Standard states: “In some situations, typically those related to specific process requirements,
alternative levels of air cleanliness may be specified on the basis of particle populations that are not
within the size range applicable to classification.”
This means continuing with one cubic metre per location.
BUT attempting this for 5.0 µm size particle could be difficult due to potential particle loss from
tubing.
Cleanroom Room size 1999 sample Revised
time per sample time
location per location
X 200 m2 3 1
Y 36 m2 6 1
Z 8 m2 12 1
No. particles @each location (or average) x (conversion factor to make one cubic
metre)
Volume of air sampled @each location
For example:
Using a particle count that counts at 28.3 litres per minute (or one cubic foot
per minute), each result would need to be multiplied by 35.3
Using a particle counter counting at 50 litres per minute, each result would be
multiplied by 20.
Individual results must be within limits per sector (unless more than one
sample per sector)
Assessment of results #2
Example: Grade B cleanroom,
There is no longer a assessed for 0.5 µm particles using
‘grand total’ for the a 1-minute counter
pass. 28.3
litres)
metre
(x 35.3)
The room is determined 1 52 1836 352,000 Pass
See:
https://www.pharmig.org.uk/en/products/publications/
or email: info@pharmig.co.uk
Dr. Tim Sandle