COMPLYING WITH INTERNATIONAL

CLEANROOM STANDARDS / GMPs

without getting ulcers

Sheesh Gulati
MeasureTest Corporation
MULTIPLICITY OF STANDARDS / GUIDELINES

F.S.209E ) Antique value?

BS 5295 )
IEST RP-CC-034.1
PDA Technical Report 13
USP 797 Sterile compounding
TGA Guidelines for Sterility testing Annex IV
PIC/S
ISO 14644
US FDA cGMP Aseptic Processing
EU GMP Annex 1

RESULT ---- Total Confusion !

 WHICH STANDARDS TO FOLLOW?
Questions to ask about Standards:
What is the source of the standard?
Is it required?
Should it be required? Can I eliminate it?
Does the standard reflect current technology?
Is it relevant to my product?
Are there newer standards that are more relevant ?
Is the standard sufficient to achieve the required
performance?
Is it realistic?
Does the standard have a potential negative effect on
the product?
Should additional standards and controls be adopted?
 FEDERAL STANDARD 209E OBSOLETE

F.S.209, for 40 years the main definition of

cleanroom classification levels, was officially
cancelled in 2001

Replaced by ISO 14644

The main differences between FS 209 and

ISO 14644 and how the new standards affect
the pharma industry is discussed in the next
few slides
 COMPARISON OF ISO 14644-1 WITH FS 209

ISO 14644-1 FED STD 209E

ISO Class English Metric
ISO 14644-1 adds
1
2 ultra-clean classes
2
ISO Class 1
3 1 M1.5
ISO Class 2
4 10 M2.5
1 very dirty class
5 100 M3.5 ISO Class 9
Total of 9 classes
6 1,000 M4.5
Counts / cubic metre
7 10,000 M5.5 Must specify room status

8 100,000 M6.5 as-built / at rest / in operation

9 - specify particle size/concentration

 ISO 14644-1 COUNT LEVELS
Airborne Particulate Cleanliness Classes (by cubic metre)

CLASS Number of Particles per Cubic Metre by Micrometre Size >=

0.1 um 0.2 um 0.3 um 0.5 um 1 um 5 um

ISO 1 10 2

ISO 2 100 24 10 4

ISO 3 1,000 237 102 35 8

ISO 4 10,000 2,370 1,020 352 83

Old Class 100> ISO 5 100,000 23,700 10,200 3,520 832 29

ISO 6 1,000,000 237,000 102,000 35,200 8,320 293

ISO 7 352,000 83,200 2,930

ISO 8 3,520,000 832,000 29,300

ISO 9 35,200,000 8,320,000 293,000

 ISO CLEANLINESS LEVELS

Points to note:

Each successively higher ISO classification allows

approximately ten times as many particles as the
previous class
The ratio of particles of size A to size B remains
approximately constant for all classes. Example:
Class 4 allows 10,200 particles 0.3 m or 3,520 m
0.5 m
Class 5 allows 102,000 particles 0.3 m or 35,200 0.5
m
 ISO 14644 CLASSES
A common mistake is to assume that, because
sizes of 0.1 micron, 0.2, 0.3, 0.5, 1 & 5 microns
are given in the Classification Table, we have to
check all these particle sizes.
The considered particle size(s) for which the
concentration will be measured, shall be agreed
upon by the customer and the supplier. Each
larger particle diameter shall be at least 1.5 times
the next smaller particle diameter.
In the pharmaceutical industry, normally one checks
0.5 and 5 microns
 25 microns particles?
The main reason 25 microns was historically required for users
is due to the BS 5295 standard [U.K.] that required monitoring
at 25 microns. In the USA, the need for 25 microns was muted.
Several other EU countries also leaned on the BS 5295
standard before ISO 14644-1 was introduced in 1999, and for a
few years after.

Note that 25 microns is specified within BS 5295 for the

equivalent of ISO Class 8 (Class 100K) and ISO Class 9.
These are "dirtier" environments. For BS 5295 Class J (ISO
Class 8), the limit was zero per cubic metre and for BS 5295
Class K (ISO Class 9), the 25 micron limit was 500/cubic metre.

For cleaner areas below ISO Class 8/Class 100K, the chart
in BS 5295 is marked as "NS" for "No Specified Limit".
 Certification: ISO 14644-1 versus FS 209E

Parameter FS 209E ISO 14644-1

2.0 litre
Minimum sample 2.83 litre
(0.07 cubic
volume (0.1 cubic foot)
foot)

Minimum sample time not specified 1 minute

Minimum number of
2 with at least 5 1 with at least 3
samples at each samples total samples total
location
Note: Typical sample volume may be larger than minimum listed above
especially for smaller size particles in very clean areas (better than ISO
Class 5 or FS 209E Class 100)
ISO 14644-1 Minimum Sample Time at 1 CFM
Time required (in minutes) at 1 cfm (28.3 lpm) flow rate with 1-minute limit imposed

0.1 um 0.2 um 0.3 um 0.5 um 1 um 5 um

ISO Class 1 70.64 353.20

ISO Class 2 7.06 29.43 70.64 176.60

ISO Class 3 1.00 2.98 6.93 20.18 88.30

ISO Class 4 1.00 1.00 1.00 2.01 8.51

ISO Class 5 1.00 1.00 1.00 1.00 1.00 24.36

ISO Class 6 1.00 1.00 1.00 1.00 1.00 2.41

ISO Class 7 1.00 1.00 1.00

ISO Class 8 1.00 1.00 1.00

ISO Class 9 1.00 1.00 1.00

 Cleanroom Certification

Initial and periodic certification

Federal Standard 209E (previously)
ISO 14644-1, -2 (now)
Max time interval for ISO Class 5 is 6 months

Three states
As-built
At Rest
Operational / Dynamic

Averaging of Data from all positions permitted

 Certification: FS209E and ISO 14644-1

Defines Cleanroom classes

Establishes minimum sampling volumes

purpose: to gather a sample volume with

theoretically at least 20 particles to help with
statistical validity of sample
Establish minimum number of points to classify area,
based on statistical criteria
Certification is also referred to as Classification or
validation or Verification
 Monitoring vs. Certification (Qualification)

EC or GMP focus: parameters during operation

dynamic or in operation
potential effect on product is critical issue
but Certification is normally done during idle time
infrequent but thorough check of the environment
as-built
at rest
Greatest concern for FDA is for viable microorganisms
Technology is not available today to measure viable
counts in real time
Non-viable counts used as a surrogate
 U.S. FDA AND ISO 14644
FDA welcomes new ISO standards as one
harmonised, base-line document on the subject of air
cleanliness classification is better than five with small
but difficult to reconcile differences
ISO documents are generic, non-industry specific,
written to meet multi-constituent industries, and often
the result is the dilution of standards to the lowest
common denominator
A company may meet the criteria of ISO 14644
but that does not mean they are complying with
cGMPs. According to FDA, classification of a clean
area is based not only on particle concentration but
also on microbiological data
FDA prohibits averaging across positions

A B
for 2 positions in ISO
Class 5 (FS 209E
Class 100) ...
125 3
119 8
120 12
364 >> 121 FDA 8 << 23
says
65
No !
 Placement of Sample Probes
No regulatory standards for
monitoring
Not controlled earlier by FS 209E or
now by ISO 14644-1, when
conducting monitoring of the process
Costly and not practical to establish
monitoring points based on the ISO
14644 formula of square root of area
in sq.metres. Risk assessment is very
important in determining where to
monitor
 ISO Class 5: ISO 14644-1 Classification
Calculations

Vial Freeze
Washing Dryer
System 5m 1

Freeze
Dryer
8m 2

5m
Calculations for Number of Points:
Freeze
Area of clean zone = 80 m Dryer
Take the SQRT (80) = 8.94 3
Rounding up to next integer = 9 sample
positions 4m

This process of selecting sample points for verification will be

compared later to the process of selecting points for the daily
monitoring of the same area.
 ISO Class 5: ISO 14644-1 Classification
Calculations

Vial Freeze
Washing Dryer
System 1
1 2 3 4 5 6

7
Freeze
Dryer
2
Calculations for Number of Points: 8

Area of clean zone = 80 m

Take the SQRT (80) = 8.94 Freeze
9
Dryer
Rounding up to next integer = 9 3
sample positions

We might place them as shown. But this does not take into
account the reality of what is in the room: entrances, exit and
machinery. So we need to adjust for these.
ISO Class 5: ISO 14644-1 Calculations
1 2 3 4 5 6 7 8 9 10

Vial Freeze
Washing Dryer
System 1

Freeze
Dryer
2
Need to adjust for equipment in room.
Under ISO 14644-1, if you sample at 10 or more
positions, you can avoid the added calculation of
the UCL (Upper Confidence Limit). Calculation of
the UCL is only mandated when the number of Freeze
positions used is between 2 and 9. Dryer
Best to sample near potential problem spots 3
which are near entrances and exits and near
operator positions.
 ISO Class 5: ISO 14644-1 Calculations
9
1
Vial 10
Freeze
Washing Dryer
8
System 1
2

11
3 4 5 6 7

Freeze
12
Dryer
2
Need to adjust for equipment in room.
Under ISO 14644-1, if you sample at 10 or more positions, 13
you can avoid the added calculation of the UCL (Upper
Confidence Limit). Calculation of the UCL is only
Freeze
mandated when the number of positions used is between
2 and 9. 14 Dryer
3
Best to sample near potential problem spots which are
near entrances and exits and near operator positions.
Here could be a distribution of sample points that would both
provide a good view of the cleanroom particulate values, and,
importantly, be defended against any regulatory challenge.
Note that there are more points suggested than the minimum
calculation.
Placement of Isokinetic Probes in a
Pharmaceutical Filling Area
for the Purpose of Monitoring

What you wont find in any book !

EU Annex I: Selecting Monitoring Positions
ISO Class 5
6
1
Vial Freeze
Washing Dryer
5
System 1
2

7
3 4

Freeze
Dryer
2
Presence of lyophilizers indicate vials may not be
fully stoppered so the holding position near 5
represents some risk 8

Position 6 provides evaluation of Grade B zone

and probably early indication of pressure balance Freeze
problems due to proximity to doors
Dryer
Positions 7 and 8 are needed because 3
loading area in front of lyophilizers should be
Grade A if product is not fully stoppered
EU Annex I: Selecting Monitoring Positions
5
1
Vial
Washing
System
2

3 4

If this were a filling operation for

which the final product remains
liquid i.e. freeze dryers were not
present, some points would not
be needed.
Placement of Isokinetic Probes in a
Life Science Manufacturing Area
(ISO Class 7 or 8)
Example: Cleanroom Area ISO Class 7 or
8 Classification

How to choose 100 ft

(30 m)
sampling points?

175 feet (53 m)

This is a large area of 30 metres by 53 metres, and rated as an ISO

Class 7 or Class 8 (FS209E Class 10K or 100K).
ISO Class 7: ISO 14644-1 verification

Entry plane (m2):

1590 m2
100 ft
SQRT (1590) = (30 m)
39.87
Minimum sample
points = 40

175 feet (53 m)

Following the ISO 14644-1 methods, we would determine that the

minimum number of sample points to carry out a formal verification
would be 40.
 ISO Class 7: ISO 14644-1 verification

Entry plane (m2):

1590 m2
100 ft
SQRT (1590) = (30 m)
39.87
Minimum sample
points = 40
6x7 grid = 42

175 feet (53 m)

But this is not an easy number to set up for a grid pattern in a rectangular area
so forty-two positions might be a better choice.
ISO Class 7: ISO 14644-1 verification

Entry plane (m2):

1590 m2
100 ft
SQRT (1590) = (30 m)
39.87
Minimum sample
points = 40
6x7 grid = 42

175 feet (53 m)

The sample positions would be at work height in the middle of each rectangle.
Example: ISO 14644-1 Calculations

1. Sum and average values at each position

2. Calculate the mean of averages
3. Result must be less than
a) the limit for the given size and
b) target room classification

If the number of points sampled is more than 1 but less than 10,
then the UCL factor must be applied:
Calculate the standard deviation
Use Students T-factor from tables
Calculate UCL
Compare to classification limit
UCL must not exceed the applicable limit
ISO Class 7: Selecting Monitoring
Positions

Work Station 1

Work Station 2

100 ft
(30 m)
Storage
Work Station 3

Work Station 4

175 feet (53 m)

In the real world, monitoring positions will be affected by the physical layout of
the area and the activities that occur within it.
Also entry and exit points should be considered.
ISO Class 7: Selecting Monitoring
Positions

Work Station 1

Work Station 2

100 ft
(30 m)
Storage
Work Station 3

Work Station 4

175 feet (53 m)

Monitoring should focus on the product exposure and vulnerability:

Where in the process is my product that most vulnerable to
contamination because of a) the length of time it sits exposed to
ambient air, b) the nature of the process step, or c) the effect
contamination might have on the next step [for example, a coating
process].
Where to monitor PIC/S recommendations

Pharmaceutical Inspection Convention, Geneva

Recommendation on the Validation of Aseptic Processing
states:
Ensure that location chosen for non-viable monitoring
reflects the worst case
For room monitoring, counts should be performed in
locations where there is most operator activity
For the filling environment,counts should be performed
adjacent to the filling zone and where components are
exposed in such a way as to detect operator activity
within these areas
 Where to monitor -- PIC recommends

Avoid monitoring in such a way that the probes

monitor the air from the HEPA filter rather than the air
immediately surrounding the critical zones
Location of the sample device should not compromise
the the laminarity of the air flow in the critical zone

FDA GMP:
Measurements should be taken with the particle
counting probe oriented in the direction of oncoming
airflow and at the sites where there is most potential
risk to the exposed product
Where to monitor?
 Where to monitor in ISO Class 5
(Class 100) / Grade filling room

General wisdom is to monitor wherever an

operator is known to breach the sterile zone
with his arm or body
Typically this may be the following 3 areas : de-
scrambler table, near the filling needle
mechanism, and the stoppering process
Some filling machines often incorporate these
functions in a more compact area and thus
only one or two positions are practical
Placement of Sample Probes
1. Sample near to exposed product
Generally near work height and exposed product
If liquid sterile fill, guidance is to sample air approaching the product within 12 (30 cm) of
exposed
Do Not measure directly above critical point
Starves the are of air
Creates turbulence
Measure to one side, close to critical location

2. Sample near to points of intervention by operators

Examples: Less than 12 inches (30

Descrambler table cm)
Filling needles
Stoppering process
Probe Sampling Positions

Isokinetic probe on an
Accumulation Turntable
provides monitoring of
rotary in-feed turntable
after sterilization zone.

Adjustable mount
(optional) allows fine
tuning of sampling
position.

Probe shown with Cap

in place
Probe Sampling Positions

Iso kinetic sampling under the

HEPA Filter in a Shrouding Machine

Isokinetic probe cups can be mounted

up to 3 metres from the counter

Each probe is connected to the counter

via a Hytrel non shedding tubing.
 EC GMP Guide Annex 1 Sterile products

Version effective September 2003 stated:

Limit of 5 micron particles for Grade A is 1
per cu.metre in operation and for Grade B
at rest
Continuous measurement system should
be used for Grade A areas (and
recommended for Grade B)
For routine testing, total sample volume
should not be <1 m for Grade A & B areas;
preferably also in Grade C areas
 REVISION OF EU-GMP GUIDE
Comments:
Research on size distribution of particles in a
cleanroom has shown that when 3500
particles of 0.5 micron are present per
cu.metre, it will contain more than one
particle of 5 micron. The well established,
and confirmed, size distribution curve used in
the ISO standard predicts 29 particles.
Therefore this stipulation was illogical and led to protests
from industry
 EU-GMP ANNEX 1 2003 REVISIONS

The use of the word "continuous" was misleading; the

right interpretation was periodic automated
sampling
Sampling intervals of 5 to 10 minutes are all right,
whereas 35 to 40 minutes would be too long.
The regulators feel that there should be zero 5
micron particles in the room, but realise that there
can be occasional "outliers". However, they expect
people to react to trends of frequent or high readings.
So, the occasional count of "1" or "2" should not stop
the line. Constant readings of say 20, should cause
an investigation.
CLASSIFICATION ACCORDING TO 0.5 MICRON

If we were just considering 0.5 micron particles, both

ISO and US FDA would permit a sampling volume of
Vs = 20/Cn.m x 1000
where Vs = Volume in litres, Cn.m = number of
particles/m3 for the relevant class
Therefore Volume of sample = 20/3520 x 1000 = 5.68
litres
With a particle counter of flow rate 1 cfm (28.3 lpm), time
taken would be less than 1 minute, so we would run the
particle counter for just 1 minute.
 1 PARTICLE OF 5 um IN QUALIFICATION

Now let us see what happens if we were to

consider 5 micron particles also.
Using the same formula, volume per location
according to ISO 14644 would be :
20/29 x 1000 = 690 litres
At sampling rate of 1 cfm (28.3 litres/mim), this
would take only 24 minutes

Which is still quite reasonable

BUT WHAT HAPPEN WITH EC GMP
LIMIT OF ONE PARTICLE OF 5 um

Volume required per location would be:

20/1 x 1000 = 20,000 litres
Time required per location at sampling rate of 28.3
l/min = 706 minutes
= 12 hours approx!
This made Certification of your cleanroom much more
expensive and time consuming.

Latest 2008 revision therefore relaxed limit to 20

particles instead of 1 particle of 5 microns/cu.metre
 EU GMP LATEST REVISION 2008
On 14th February 2008, the European Commission updated
Volume 4 of EU Guidelines to the GMPs for medicinal products
for Human and Veterinary use
This revised Annex 1 will come into operation on 1st. March 2009
except for the provisions on capping of freeze-dried vials,
which should be implemented by 1st. March 2010.
It clearly outlines three phases that need to be performed:
Certification: Each cleanroom and clean air device should first be
classified
Monitoring: the cleanroom should then be monitored to verify that
conditions are being maintained relative to product quality
Data Review: Ensure that the data accrued from the monitoring be
reviewed in the light of risk to finished product quality.
 EU GMP LATEST REVISION 2008
The maximum permitted airborne particle concentration for each
grade is given in the following table
Maximum permitted number of particles per m3 equal to
or greater than the tabulated size
At rest In operation

Grade 0.5 m 5.0m 0.5 m 5.0m

A 3 520 20 3 520 20
(ISO 5 = 29)
B 3 520 29 352 000 2 900
(ISO 7 =2930)
C 352 000 2 900 3 520 000 29 000
(ISO 7 = 2930) (ISO 8 = 29,300)
D 3 520 000 29 000 Not Not defined
(ISO 8= defined
29,300)
 EU GMP 2008 REVISION
Continuous Discontinued?
The new revision does not specifically mention Continuous
Measurement Systems are mandatory, but it is implied :
For Grade A zones, particle monitoring should be
undertaken for the full duration of critical
processing, including equipment assembly

The Grade A zone should be monitored at such a

frequency and with suitable sample size that all
interventions, transient events and any system
deterioration would be captured and alarms
triggered if alert limits are exceeded
 EU Annex 1: March 2009 Changes
At Rest In Operation

Grade Maximum permitted number of particles/m3 equal to or above

Existing
0.5 m 5 m 0.5 m 5 m Effective Sept. 2003

A 3 500 1 3 500 1 5 m limits for Grade A & B

0 1 per cubic meter
B 3 500 1 350 000 2 000
C 350 000 2 000 3 500 000 20 000

D 3 5000 000 20 000 not defined not defined

At Rest In Operation
3
Grade Maximum permitted number of particles/m equal to or above

0.5 m 5 m 0.5 m 5 m
New
A 3 520 20 3 520 20 Effective Mar. 2009

B 3520 29 352 000 2 900

5 m limits for Grade A
C 352 000 2 900 3 520 000 29 000
1 20 per cubic meter
D 3 520 000 29 000 not defined not defined
 Why measure 5 um particles ?

EU inspectors maintain that large particles are

potential carriers (hitch-hikers), of or are, viable
organisms themselves.
If these particles are present in an aseptic
environment, they represent an increased risk of
contamination of the sterile product.
Large particles do not transport well in tubing runs
exceeding 3 metres (10 feet). Keep tubing runs
from the sample site to the particle counter as
short as possible to avoid particle loss.
5 micron counts can be an indicator of:
Problems with the physical plant
Problems with personnel and procedures
 SEQUENTIAL MANIFOLD SYSTEMS

1. Although manifold type sequential monitoring systems

are not banned, it will be difficult to justify the use of
manifolds after 1st.September 2003. Evidence might be
expected that manifold systems had documented
efficiency at larger particles the length of tubing and
the radii of any bends in the tubing must be considered
in the context of particle losses in the tubing.

2. I believe that our intention was not to ban cyclical sampling.

By continuous we meant throughout the filling run. But we would
expect, again, for the sampling regime to be documented; the
rationale explained and justified. if you had a cyclical manifold
and it was sampling in the Grade A zone once every 45 minutes,
then you might have a bit of a problem justifying that. If it is
sampling, for example, every 5 minutes, the justification would be
very much easier to write.
Paul Hargreaves, MHRA
Why EC GMP doesnt like Manifolds

Tubing Transport Loss

100
90
80 0,1
% 70 0,5
L 60 0,7
o 50 1
s 40 3
s 30 5
20 10
10
0
2 7 13 19 26 30
Length of tubing (meters)
EU Annex 1 2008 Revision Summary
For verification (classification of room)
Section 3: (Enhanced definition of at rest)
The at-rest state is the condition where the installation is installed
and operating, complete with production equipment but with no
operating personnel present.
Section 4:
Classification should be clearly differentiated from operational
process environmental monitoring.
Section 5:
For classification purposes EN/ISO 14644-1 methodology defines
both the minimum number of sample locations and the [minimum]
sample size based on the class limit of the largest considered
particle size and the method of evaluation of the data collected
For classification purposes in Grade A zones, a minimum
sample volume of 1 m3 should be taken per sample position.
 EU Annex 1 2008 Summary
For monitoring (for example, with an FMS system)
there is no minimum volume or time period for each sample
In Annex I, the only statements are to
encourage a continuous sampling system for the Grade A areas
encourage a continuous sampling system for the Grade B areas,
although not so necessary as for Grade A
indicate that the sample rate can be different than that used to
qualify the area

Section 12 states:
"It is not necessary for the sample volume to be the same as that used
for formal classification of clean rooms and clean air devices.
i.e. you do not need to sample minimum 1 cu metre during monitoring
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Increased sampling frequency of low air volume is
preferable to high air volume at low frequency
In other words, 35 readings of 1 minute at 1 cfm are
preferable to 1 reading of 35 minutes
Action limit for 1 cu.metre
Single readings: If all readings are below 1/35 of the
limjt, the limit will never be exceeded
Multiple readings: If some of the single readings exceed
the 1/35 of cu.metre limit, it has to be checked
whether the result of sampling 1 cu.metre air volume
would have exceeded the limit
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Action limit for 1 cft readings:

Readings below 1/35 of the 1 m3 requirement

are acceptable
100 counts/cft for 0.5 micron
0 counts/cft for 5 microns

Any reading exceeding 1 m3 requirement is not

acceptable:
3521 counts for 0.5 micron &
21 counts for 5 micron particles
SUGGESTED PARTICLE MONITORING
REGIME
Sample size and frequency should be based
on likelihood of finding a contamination
event
A long sample time --and hence large volume
could allow a short term even to be diluted
by a low subsequent count rate
A short sample time alone might cause you
to think a false or spurious count is a real
major contamination event
Correlation with media fill data? Usually
unlikely
Correlation with an activity ? Highly likely
SUGGESTED PARTICLE MONITORING REGIME
Considering the >= 5 micron particles,
EU GMP limit is 20 per cu metre
1 cu metre takes about 36 minutes to sample at I cfm
i.e. 28.3 lpm
If we wait 36 minutes before evaluating a sample, we
could miss an event
If we only look at a small sample of 1 cft (28.3 litre),
then 1 real or false particle would imply 35/cu metre
= FAILING the 20 limit
So we should look simultaneously at each 28.3 litre
sample and a cumulative 1000 litre (1 m3) sample
 EU GMP -- VIAL CAPPING ISSUES
Relates to freeze dried products, liquid and
solid fill applications
Concerns were raised by inspectors when
seeing mis-placed stoppers re-seated by
hand or even stoppered by hand when
missing
Partially stoppered freeze dried vials
maintained under Grade A conditions at all
times
Non-freeze dried vials protected with a
Grade A air supply. Is protected the same
as maintained?
 EU GMP VIAL CAPPING
Containers should be closed by appropriately
validated methods. Containers closed by fusion
e.g. glass or plastic ampoules should be subject
to 100% integrity testing. Samples of other
containers should be checked for integrity
according to appropriate procedures

Partially stoppered freeze drying vials should be

maintained under grade A conditions at all times
until the stopper is fully inserted

The container closure system for aseptically filled

vials is not fully integral until the aluminium cap has
been crimped into place on the stoppered vial
 EU GMP VIAL CAPPING
As the equipment used to crimp vial caps can generate
large quantities of non viable particulates, the
equipment should be located at a separate station
equipped with adequate air extraction

Vial capping can be undertaken as an aseptic process

using sterilized caps or as a clean process outside the
aseptic core. Where this latter approach is adopted,
vials should be protected by Grade A conditions up to
the point of leaving the aseptic processing area, and
thereafter stoppered vials should be protected with a
Grade A air supply until the cap has been crimped.
 EU GMP VIAL CAPPING
Note that a Grade A air supply is differentiated
from a Grade A environment

Vials with missing or displaced stoppers should

be rejected prior to capping. Where human
intervention is requires at the capping station,
appropriate technology should be used to
prevent direct contact with the vials and to
minimize microbial contamination

This part of Annex 1 will be effective from

1st.March 2010
Interference from capping operation

Unidirectional air shower

Higher sample
probe for
monitoring during
capping operation

30 tha s
s
Sample probe to 5 n
Le

m
m
demonstrate air
quality before
capping process
Interpreting EU GMP Annex 1

the PHSS Best Practice Guide

(Pharmaceutical & Healthcare Sciences Society,
U.K.)
 Advice on Best Practice for
Cleanroom Monitoring
None in Annex 1
What system to use?
Where to locate monitoring points?
How do we deal with 5m counts?
1m3 volume during manufacture?
Powder fill applications?

FDA cGMP
areas where product is at most potential risk
not more than 1 foot away from the work site
 Advice on Best Practice for
Cleanroom Monitoring
What was lacking was practical advice on how to implement these
continuous monitoring systems. There is none in Annex 1 and
ISO14644 is for room classification only.
Continuous what, for example, does the word continuous mean?
Monitoring where should we locate the monitoring points and how
many should there be?
5micron what do we do if we see 5micron counts. How many are
acceptable before we initiate and action limit
1 cubic metre it is not clear as to whether we should try to sample a
complete cubic meter during each manufacturing batch. Some
aseptic manipulations are complete in a matter of minutes and it
currently takes at least 20 minutes to capture 1cubic metre of air
with a modern counter.
Powder are we really supposed to monitor for particles during a
powder fill?
Scope & Aims of PHSS Special Interest
Group

Scope:
Cleanroom non-viable air particle monitoring
EU GMP Annex 1

Aims:
Collate best practice from Industry, Healthcare and regulatory
bodies
Publish monograph :

Best Practice for Particle Monitoring

in
Pharmaceutical Cleanrooms
Best Practice Document the team

AstraZeneca
Bio Products
Boehringer-Ingelheim
Boots Contract Manufacturing
Cardinal Health
GlaxoSmithKline
Hach Ultra Analytics
Ipsen Biopharm
Particle Measurement Techniques
Wyeth
MHRA Regulatory Inspectors (EMeA)
 Best Practice Document Contents
Changes to EU GMP Annex 1 published 2008, live March
2009
System Design
Operations
Maintenance and Cleaning
Training
Appendix A Worked example
Appendix B Manifold and Remote Particle Monitoring
Systems
Appendix C Examples of particle loss in transport tubing
Appendix D Isokinetic probes
Appendix E Validation and risk assessment standards
and guidelines
 UK PHSS Best Practice monitoring
Grade A areas monitored continuously using dedicated particle
counters.

Grade B areas (background for a Grade A) use dedicated particle

counters.

Other Grade B areas and Grade C areas may be monitored by manifold

systems to check that they are under control.
(Note: There are no limits for the in operation state in Grade D areas.)

Corridors and change areas may be checked on a routine basis using

portable particle counters or monitored using manifold systems.

Enhanced monitoring should be provided in certain Grade C and D areas,

for example in biologicals sites where low grade areas can
potentially contribute a significant bioburden (to the point of
sterility failure).
Appendix B dedicated counter Grade A

Vial
Sterilizing
Tunnel

Key
Remote
Counter

Vacuum
Tubing,
Power
& Data

Central Central
Software Vacuum
System Pump
Appendix B dedicated counter Grade A

Vial
Sterilizing 0.5

Tunnel
0.5

Key

Remote 0.5

Counter 0.5 0.5

(Built-in
Pump)

Power
& Data

Vacuum
Tubing
Central
Software
System
Appendix B Manifold for Grade B & C

Vial
Sterilising
Tunnel

Key
Sample
Probe

Vacuum
Tubing

Particle Controller Manifold

Counter
 FDA ASEPTIC PROCESSING
CGMP GUIDANCE

After over 15 years, US FDA published on 27th. September

2002, a Concept Paper entitled Sterile Drug Products
produced by Aseptic Processing & and invited comments
Subsequently issued as Draft GMP guidance in August 2003
and final CGMP in September 2004
New topics include guidance for personnel qualification,
cleanroom classifications under dynamic conditions,
environmental monitoring, isolators, blow-fill-seal systems
Appendix 1 reiterates FDAs view that isolators should not
be located in unclassified rooms and suggests Class
100,000 (ISO Class 8) background
 FDA ASEPTIC PROCESSING
CGMP GUIDANCE

Air classification given in Table 1 of Buildings &

Facilities only gives number of particles of 0.5 micron
and larger per cft/cu.metre. Also dynamic state only.
In this respect it differs from EU GMP as no mention
is made of 5 micron particles
Regular monitoring should be performed during each
shift
Non-viable particulate monitoring with a remote
counting system is generally less invasive than the
use of portable particle counting units and provide the
most comprehensive data
 FDA ASEPTIC PROCESSING
CGMP GUIDANCE

Air changes - For Class 100,000 (ISO Class 8)

supporting rooms, at least 20 air changes per
hours is typically acceptable, for higher
cleanliness classes significantly higher air
change rates
Air velocity: air in critical areas should be
supplied at a velocity sufficient to sweep
particulate matter away from the filling/closing
area and maintain laminarity.
A velocity of 90 to 100 ft/min +-20% is
recommended
 90 to 100 ft/min air velocity?

The 90 to 100 fpm +/- 20% value should be a

"guideline value" (in MCA terminology) or
"informative" (in ISO terminology).
The magic of 90 fpm has been known to be a
fallacy within the cleanroom industry for over 25
years.
This value is based on a simple calculation that a 5
um particle would stay airborne (settle less than 2
feet) over a distance of 20 feet in a horizontal flow
cleanroom.
The true test is airflow pattern testing
 U.S.FDA ASEPTIC PROCESSING
CGMP

Differential pressure should be monitored continuously

throughout each shift and frequently recorded
We recommend conducting non-viable particle monitoring
with a remote counting system
Airflow velocities are measured 6 inches from the filter face
or at a defined distance proximal to the work surface, for
each HEPA filter
Samples from Class 100 (ISO Class 5) environments
should normally yield no microbiological contaminants
 FDA GMP -WEAKNESSES

Dynamic Classification Expected

the final room or area classification should be
derived from data generated under dynamic
conditions.
Comments
This is very difficult in practice, and facilities are
normally classified under static conditions as per ISO
14644.Various factors outside the control of
cleanroom contractors make classification in
operational mode difficult such as gowning practice,
microbial control, etc.
 FDA GMP -WEAKNESSES
Sterility Expectations
Air monitoring of critical areas should normally yield no
microbiological contaminants
also Samples from Class 100 environments should
normally yield no microbiological contaminants

Comments:
Attaining a sterile state in an aseptic processing
facility is impossible. Personnel are always present in
manned cleanrooms performing various activities
including microbial sampling. Detection of micro-
organisms occasionally is inevitable and need not be
a cause for action against product.
 AIR CHANGES
Describing airflow in terms of air changes per hour is common
for non-unidirectional flow rooms (ISO classes 6 through 9)
and high-bay installations.
Since the airflow in these rooms is non-uniform, attempting to
directly measure the average air velocity is not feasible. The
average velocity may be calculated, however, using
volumetric measurements from the terminal filters. This
velocity is then converted into an equivalent room air
changes per hour (AC/H).
It is worth noting that in the UK GMP ('Orange Guide')
the room air change requirements have been removed
in the latest edition.
 EU Annex 1 vs. FDA Guideline
EU Annex 1 FDA Guideline

Sizes 5 micron 0.5 micron

monitored 0.5 micron

Room Grades A, B, C, D Critical = A

Classes Grade B as Controlled = C, D
surrounding Grade A

States to be At rest In operation

monitored In operation

FDA does not require classification of final stage of changing

room to be the same as the room into which it leads
 FDA CGMP -- HEPA FILTER TESTING

HEPA filter integrity testing should be performed

twice a year
DOP /PAO challenge and scanning with aerosol
photometer is recommended (DOP not banned)
Concentration of poly-dispersed aerosol should be
appropriate for the accuracy of the photometer
(previously FDA had suggested 80 to 100 ug/L which
was too high. Normally 20ug/L sufficient)
 EN 1822 - European Standard for Filters

HEPA Filter efficiency tested at MPPS

Leak Testing also at MPPS
Efficiency as low as 85% rated as HEPA
Particle Counter and CNC only, no photometers
A suitable 0.1 micron sensitivity particle counter
would also require a dilutor and the total cost
would be around $ 20,000 i.e. double that of
a photometer
 TIME TO CHANGE
OLD TERMINOLOGY NEW TERMINOLOGY
Laminar flow Unidirectional Flow

Clean Room Cleanroom (one word)

Class 100, Class 1000 Class 5, Class 6 etc

Micron Micrometre
Static/dynamic At rest/operational
Air pattern/smoke Airflow visualisation