cleanroom standards
COMPLYING WITH INTERNATIONAL
CLEANROOMSTANDARDS / GMPs
without getting ulcers
SheeshGulatiMeasureTestCorporation
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 cGMPAseptic Processing•EU GMP Annex 1RESULT ----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
cleanroomclassification 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 pharmaindustry is discussed in the next
few slides
COMPARISON OF ISO 14644-1 WITH FS 209
ISO 14644-1FED STD 209E1231M1.5410M2.55100M3.561,000M4.59ISO ClassEnglishMetric710,000M5.58100,000M6.5
Text Box: ISO 14644-1 adds
2 “ultra-clean” classes
ISO Class 1
ISO Class 2
1 “very dirty” class
ISO Class 9
Total of 9 classes
Counts / cubic metre
Must specify room status
“as-built” / “at rest” / “in operation”
- specify particle size/concentr
ISO 14644-1 adds•2 “ultra-clean”classes–ISO Class 1–ISO Class 2•1 “very dirty”class–ISO Class 9Total of 9 classesCounts / cubic metreMust specify room status•“as-built”/ “at rest”/ “in operation”
-specify particle size/concentration
Text Box: ISO 14644-1 COUNT LEVELS
ISO 14644-1 COUNT LEVELSAirborne Particulate Cleanliness Classes (by cubic metre)
CLASSNumber of Particles per Cubic Metre by MicrometreSize >=
Old Class 100>
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 µm or 3,520 µm
=0.5 µm
Class 5 allows 102,000 particles =0.3 µm or 35,200 =0.5
µm
ISO 14644 CLASSES
A common mistakeis 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
Minimum sample timenot specified1 minuteMinimum number of
samples at each
locationISO 14644-12.0 litre(0.07 cubic
foot)
1 with at least 3
samples totalParameterMinimum sample
volume2 with at least 5
samples totalFS 209E2.83 litre(0.1 cubic foot)
Text Box: 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)
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 imposed0.1 um0.2 um0.3 um0.5 um1 um5 umISO Class 170.64353.20ISO Class 27.0629.4370.64176.60ISO Class 31.002.986.9320.1888.30ISO Class 41.001.001.002.018.51ISO Class 51.001.001.001.001.0024.36ISO Class 61.001.001.001.001.002.41ISO Class 71.001.001.00ISO Class 81.001.001.00ISO Class 91.001.001.00
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”
125119120364>>121“B”
3812238 <<
65FDA
says“No !”
for 2 positions in ISO
Class 5 (FS 209E
Class 100) ...
Placement of Sample Probes
Text Box: 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
•No regulatory standards for
monitoring•Not controlled earlier by FS 209E or
now by ISO 14644-1, when
conducting monitoringof 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
Calculations for Number of Points:
Area of clean zone = 80 m²
Take the SQRT (80) = 8.94
Rounding up to next integer = 9 sample
positions
8 m5 m5 mFreeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
4 mThis process of selecting sample points for verificationwill be
compared later to the process of selecting points for the daily
monitoringof the same area.
ISO Class 5: ISO 14644-1 Classification
Calculations
Calculations for Number of Points:
Area of clean zone = 80 m²
Take the SQRT (80) = 8.94Rounding up to next integer = 9
sample positions123456789Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
Text Box: 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.
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
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
positions used is between 2 and 9.
Best to sample near potential problem spots
which are near entrances and exits and near
operator positions.
Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
Oval: 1
1
Oval: 2
2
Oval: 3
3
Oval: 4
4
Oval: 5
5
Oval: 6
6
Oval: 7
7
Oval: 8
8
Oval: 9
9
Oval: 10
10
ISO Class 5: ISO 14644-1 Calculations
Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System1234567891011121314Need 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 positions used is between
2 and 9.
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 cleanroomparticulate 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 won’t find in any book !
EU Annex I: Selecting Monitoring Positions
ISO Class 5
Presence of lyophilizers indicate vials may not be
fully stoppered so the holding position near “5”
represents some riskPosition “6”provides evaluation of Grade B zone
and probably early indication of pressure balance
problems due to proximity to doorsPositions “7”and “8”are needed because
loading area in front of lyophilizers should be
Grade A if product is not fully stoppered12345678Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
EU Annex I: Selecting Monitoring Positions
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.
12345Vial
Washing
System
Placement of Isokinetic Probes in a
Life Science Manufacturing Area
(ISO Class 7 or 8)
Example: Cleanroom Area ISO Class 7 or
8 Classification
100 ft
(30 m)How to choose
sampling points?
175 feet (53 m)
Text Box: 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).
This is a large area of 30 metresby 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
SQRT (1590) =
39.87
Minimum sample
points = 40
100 ft
(30 m)
175 feet (53 m)
Text Box: 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.
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
SQRT (1590) =
39.87
Minimum sample
points = 40
6x7 grid = 42
100 ft
(30 m)
175 feet (53 m)
Text Box: 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.
But this is not an easy number to set up for a grid pattern in arectangular area
so forty-two positions might be a better choice.
ISO Class 7: ISO 14644-1 verification
Entry plane (m2):
1590 m2
SQRT (1590) =
39.87
Minimum sample
points = 40
6x7 grid = 42
100 ft
(30 m)
175 feet (53 m)
Text Box: The sample positions would be at work height in the middle of each rectangle.
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 Student’s T-factor from tables
•Calculate UCL
•Compare to classification limit
•UCL must not exceed the applicable limit
ISO Class 7: Selecting MonitoringPositions
100 ft
(30 m)
Work Station 1Work Station 2Work Station 3Work Station 4Storage
175 feet (53 m)
Text Box: 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.
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 MonitoringPositions
100 ft
(30 m)
Work Station 1Work Station 2Work Station 3Work Station 4Storage
175 feet (53 m)
Text Box: 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
Monitoringshould 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 effectcontamination 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 laminarityof 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 bodyTypically this may be the following 3 areas : de-
scrambler table, near the filling needle
mechanism, and the stopperingprocessSome filling machines often incorporate these
functions in a more compact area and thus
only one or two positions are practical
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
Placement of Sample Probes
Less than 12 inches(30
cm)
2. Sample near to points of intervention by operatorsExamples:
•Descrambler table•Filling needles•Stoppering process
Probe Sampling Positions
Isokineticprobe 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
Isokinetic sampling under the
HEPA Filter in a Shrouding Machine
Isokineticprobe cups can be mounted
up to 3 metres from the counter
Each probe is connected to the counter
via a Hytrelnon 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 m³for Grade A & B areas;
preferably also in Grade C areas
REVISION OF EU-GMP GUIDE
Comments:
Research on size distribution of particles in a
cleanroomhas 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 cleanroommuch 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 14thFebruary 2008, the European Commission updated
Volume 4 of EU Guidelines to the GMPsfor 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 cleanroomand clean air device should first be
classified
Monitoring:the cleanroomshould 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
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
Existing
Effective Sept. 2003
GradeMaximum permitted number of particles/m3 equal to or above0.5 µm5 µm0.5 µm5 µmA3 50013 5001B3 5001350 0002 000C350 0002 0003 500 00020 000D3 5000 00020 000not definednot definedAt RestIn Operation
At Rest In Operation
Grade Maximum permitted number of particles/m3 equal to or above
0.5 µm 5 µm 0.5 µm 5 µm
A 3 520 20 3 520 20
B 3520 29 352 000 2 900
C 352 000 2 900 3 520 000 29 000
D 3 520 000 29 000 not defined not defined
NewEffective Mar. 20095 µm limits for Grade A & B0 ..1 per cubic meter5 µm limits for Grade A1 ..20 per cubic meter
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 doesn’t like Manifolds
Tubing Transport LossTubing Transport Loss01020304050607080901002713192630Length of tubing (meters)
%
Loss0,1 µ0,5 µ0,7 µ1 µ3 µ5 µ10 µ
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:
“Classificationshould be clearly differentiated from operational
process environmental monitoring.”
Section 5:
“For classificationpurposes 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 m3should 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
•encouragea continuous sampling system for the Grade A areas
•encouragea continuous sampling system for the GradeB areas,
although not so necessary as for Grade A
•indicatethat 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 metreduring monitoring
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Increased sampling frequency of low air volume is
preferable to high air volume at low frequencyIn other words, 35 readings of 1 minute at 1 cfmare
preferable to 1 reading of 35 minutesAction limit for 1 cu.metreSingle readings: If all readings are below 1/35 of the
limjt, the limit will never be exceededMultiple readings: If some of the single readings exceed
the 1/35 of cu.metrelimit, it has to be checked
whether the result of sampling 1 cu.metreair volume
would have exceeded the limit
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Action limit for 1 cftreadings:
Readings below 1/35 of the 1 m3 requirement
are acceptable
•100 counts/cftfor 0.5 micron
•0 counts/cftfor 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 metretakes about 36 minutes to sample at I cfmi.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 litresample 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 stopperedby hand when
missing
•Partially stopperedfreeze 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 stopperedfreeze 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 aluminiumcap has
been crimped into place on the stopperedvial
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 stopperedvials 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
Sample probe to
demonstrate air
quality before
capping processLess
than305 mmUnidirectional air showerHigher sample
probe for
monitoring during
capping operation
Text Box: Interpreting EU GMP Annex 1 –
the PHSS Best Practice Guide
(Pharmaceutical & Healthcare Sciences Society, U.K.)
the PHSS Best Practice Guide
Advice on Best Practice for
CleanroomMonitoring
None in Annex 1
–What system to use?
–Where to locate monitoring points?
–How do we deal with 5µm counts?
–1m3volume 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
CleanroomMonitoring
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 itcurrently 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 Aareas monitored continuously using dedicated particle
counters.
Grade Bareas (background for a Grade A) use dedicated particle
counters.
Other Grade B areas and Grade Careas may be monitored by manifoldsystems to check that they are under control.
(Note: There are no limits for the ‘in operation’state in Grade D areas.)
Corridors and change areasmay be checked on a routine basis using
portable particle countersor monitored using manifold systems.
Enhanced monitoringshould 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
VialSterilizing
TunnelCentralVacuumPumpKeyRemoteCounterVacuumTubing,
Power& DataCentralSoftwareSystem
Appendix B –dedicated counter Grade A
VialSterilizing
TunnelKeyRemoteCounter(Built-in
Pump)
Power& DataVacuumTubingCentralSoftwareSystem0.5µ0.5µ0.5µ0.5µ0.5µ
Appendix B –Manifold for Grade B & C
VialSterilisingTunnelParticleCounterControllerManifoldKeySampleProbeVacuumTubingVialSterilisingTunnelParticleCounterControllerMnifoldKeySampleProbeVacuumTubingVialSterilisingTunnelParticleCounterControllerManifoldKeySampleProbeVacuumTubing
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,
cleanroomclassifications under dynamic conditions,
environmental monitoring, isolators, blow-fill-seal systems
Appendix 1 reiterates FDA’s 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 cleanroomindustry 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”.
CommentsThis is very difficult in practice, and facilities are
normally classified under static conditions as per ISO
14644.Various factors outside the control of
cleanroomcontractors 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 nomicrobiological 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 cleanroomsperforming 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
In operationAt restIn operationStates to be
monitoredCritical = AControlled = C, DGrades A, B, C, DGrade B as
surrounding Grade ARoom
Classes0.5 micron5 micron0.5 micronSizes
monitoredFDA GuidelineEU Annex 1
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 MPPSLeak Testing also at MPPSEfficiency as low as 85% rated as HEPAParticle Counter and CNC only, no photometersA 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
CLEANROOMSTANDARDS / GMPs
without getting ulcers
SheeshGulatiMeasureTestCorporation
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 cGMPAseptic Processing•EU GMP Annex 1RESULT ----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
cleanroomclassification 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 pharmaindustry is discussed in the next
few slides
COMPARISON OF ISO 14644-1 WITH FS 209
ISO 14644-1FED STD 209E1231M1.5410M2.55100M3.561,000M4.59ISO ClassEnglishMetric710,000M5.58100,000M6.5
Text Box: ISO 14644-1 adds
2 “ultra-clean” classes
ISO Class 1
ISO Class 2
1 “very dirty” class
ISO Class 9
Total of 9 classes
Counts / cubic metre
Must specify room status
“as-built” / “at rest” / “in operation”
- specify particle size/concentr
ISO 14644-1 adds•2 “ultra-clean”classes–ISO Class 1–ISO Class 2•1 “very dirty”class–ISO Class 9Total of 9 classesCounts / cubic metreMust specify room status•“as-built”/ “at rest”/ “in operation”
-specify particle size/concentration
Text Box: ISO 14644-1 COUNT LEVELS
ISO 14644-1 COUNT LEVELSAirborne Particulate Cleanliness Classes (by cubic metre)
CLASSNumber of Particles per Cubic Metre by MicrometreSize >=
Old Class 100>
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 µm or 3,520 µm
=0.5 µm
Class 5 allows 102,000 particles =0.3 µm or 35,200 =0.5
µm
ISO 14644 CLASSES
A common mistakeis 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
Minimum sample timenot specified1 minuteMinimum number of
samples at each
locationISO 14644-12.0 litre(0.07 cubic
foot)
1 with at least 3
samples totalParameterMinimum sample
volume2 with at least 5
samples totalFS 209E2.83 litre(0.1 cubic foot)
Text Box: 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)
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 imposed0.1 um0.2 um0.3 um0.5 um1 um5 umISO Class 170.64353.20ISO Class 27.0629.4370.64176.60ISO Class 31.002.986.9320.1888.30ISO Class 41.001.001.002.018.51ISO Class 51.001.001.001.001.0024.36ISO Class 61.001.001.001.001.002.41ISO Class 71.001.001.00ISO Class 81.001.001.00ISO Class 91.001.001.00
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”
125119120364>>121“B”
3812238 <<
65FDA
says“No !”
for 2 positions in ISO
Class 5 (FS 209E
Class 100) ...
Placement of Sample Probes
Text Box: 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
•No regulatory standards for
monitoring•Not controlled earlier by FS 209E or
now by ISO 14644-1, when
conducting monitoringof 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
Calculations for Number of Points:
Area of clean zone = 80 m²
Take the SQRT (80) = 8.94
Rounding up to next integer = 9 sample
positions
8 m5 m5 mFreeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
4 mThis process of selecting sample points for verificationwill be
compared later to the process of selecting points for the daily
monitoringof the same area.
ISO Class 5: ISO 14644-1 Classification
Calculations
Calculations for Number of Points:
Area of clean zone = 80 m²
Take the SQRT (80) = 8.94Rounding up to next integer = 9
sample positions123456789Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
Text Box: 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.
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
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
positions used is between 2 and 9.
Best to sample near potential problem spots
which are near entrances and exits and near
operator positions.
Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
Oval: 1
1
Oval: 2
2
Oval: 3
3
Oval: 4
4
Oval: 5
5
Oval: 6
6
Oval: 7
7
Oval: 8
8
Oval: 9
9
Oval: 10
10
ISO Class 5: ISO 14644-1 Calculations
Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System1234567891011121314Need 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 positions used is between
2 and 9.
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 cleanroomparticulate 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 won’t find in any book !
EU Annex I: Selecting Monitoring Positions
ISO Class 5
Presence of lyophilizers indicate vials may not be
fully stoppered so the holding position near “5”
represents some riskPosition “6”provides evaluation of Grade B zone
and probably early indication of pressure balance
problems due to proximity to doorsPositions “7”and “8”are needed because
loading area in front of lyophilizers should be
Grade A if product is not fully stoppered12345678Freeze
Dryer1Freeze
Dryer2Freeze
Dryer3Vial
Washing
System
EU Annex I: Selecting Monitoring Positions
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.
12345Vial
Washing
System
Placement of Isokinetic Probes in a
Life Science Manufacturing Area
(ISO Class 7 or 8)
Example: Cleanroom Area ISO Class 7 or
8 Classification
100 ft
(30 m)How to choose
sampling points?
175 feet (53 m)
Text Box: 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).
This is a large area of 30 metresby 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
SQRT (1590) =
39.87
Minimum sample
points = 40
100 ft
(30 m)
175 feet (53 m)
Text Box: 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.
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
SQRT (1590) =
39.87
Minimum sample
points = 40
6x7 grid = 42
100 ft
(30 m)
175 feet (53 m)
Text Box: 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.
But this is not an easy number to set up for a grid pattern in arectangular area
so forty-two positions might be a better choice.
ISO Class 7: ISO 14644-1 verification
Entry plane (m2):
1590 m2
SQRT (1590) =
39.87
Minimum sample
points = 40
6x7 grid = 42
100 ft
(30 m)
175 feet (53 m)
Text Box: The sample positions would be at work height in the middle of each rectangle.
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 Student’s T-factor from tables
•Calculate UCL
•Compare to classification limit
•UCL must not exceed the applicable limit
ISO Class 7: Selecting MonitoringPositions
100 ft
(30 m)
Work Station 1Work Station 2Work Station 3Work Station 4Storage
175 feet (53 m)
Text Box: 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.
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 MonitoringPositions
100 ft
(30 m)
Work Station 1Work Station 2Work Station 3Work Station 4Storage
175 feet (53 m)
Text Box: 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
Monitoringshould 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 effectcontamination 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 laminarityof 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 bodyTypically this may be the following 3 areas : de-
scrambler table, near the filling needle
mechanism, and the stopperingprocessSome filling machines often incorporate these
functions in a more compact area and thus
only one or two positions are practical
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
Placement of Sample Probes
Less than 12 inches(30
cm)
2. Sample near to points of intervention by operatorsExamples:
•Descrambler table•Filling needles•Stoppering process
Probe Sampling Positions
Isokineticprobe 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
Isokinetic sampling under the
HEPA Filter in a Shrouding Machine
Isokineticprobe cups can be mounted
up to 3 metres from the counter
Each probe is connected to the counter
via a Hytrelnon 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 m³for Grade A & B areas;
preferably also in Grade C areas
REVISION OF EU-GMP GUIDE
Comments:
Research on size distribution of particles in a
cleanroomhas 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 cleanroommuch 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 14thFebruary 2008, the European Commission updated
Volume 4 of EU Guidelines to the GMPsfor 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 cleanroomand clean air device should first be
classified
Monitoring:the cleanroomshould 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
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
Existing
Effective Sept. 2003
GradeMaximum permitted number of particles/m3 equal to or above0.5 µm5 µm0.5 µm5 µmA3 50013 5001B3 5001350 0002 000C350 0002 0003 500 00020 000D3 5000 00020 000not definednot definedAt RestIn Operation
At Rest In Operation
Grade Maximum permitted number of particles/m3 equal to or above
0.5 µm 5 µm 0.5 µm 5 µm
A 3 520 20 3 520 20
B 3520 29 352 000 2 900
C 352 000 2 900 3 520 000 29 000
D 3 520 000 29 000 not defined not defined
NewEffective Mar. 20095 µm limits for Grade A & B0 ..1 per cubic meter5 µm limits for Grade A1 ..20 per cubic meter
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 doesn’t like Manifolds
Tubing Transport LossTubing Transport Loss01020304050607080901002713192630Length of tubing (meters)
%
Loss0,1 µ0,5 µ0,7 µ1 µ3 µ5 µ10 µ
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:
“Classificationshould be clearly differentiated from operational
process environmental monitoring.”
Section 5:
“For classificationpurposes 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 m3should 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
•encouragea continuous sampling system for the Grade A areas
•encouragea continuous sampling system for the GradeB areas,
although not so necessary as for Grade A
•indicatethat 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 metreduring monitoring
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Increased sampling frequency of low air volume is
preferable to high air volume at low frequencyIn other words, 35 readings of 1 minute at 1 cfmare
preferable to 1 reading of 35 minutesAction limit for 1 cu.metreSingle readings: If all readings are below 1/35 of the
limjt, the limit will never be exceededMultiple readings: If some of the single readings exceed
the 1/35 of cu.metrelimit, it has to be checked
whether the result of sampling 1 cu.metreair volume
would have exceeded the limit
DEALING WITH 1 CU.METRE REQUIREMENT
DURING CLASSIFICATION
Action limit for 1 cftreadings:
Readings below 1/35 of the 1 m3 requirement
are acceptable
•100 counts/cftfor 0.5 micron
•0 counts/cftfor 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 metretakes about 36 minutes to sample at I cfmi.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 litresample 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 stopperedby hand when
missing
•Partially stopperedfreeze 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 stopperedfreeze 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 aluminiumcap has
been crimped into place on the stopperedvial
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 stopperedvials 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
Sample probe to
demonstrate air
quality before
capping processLess
than305 mmUnidirectional air showerHigher sample
probe for
monitoring during
capping operation
Text Box: Interpreting EU GMP Annex 1 –
the PHSS Best Practice Guide
(Pharmaceutical & Healthcare Sciences Society, U.K.)
the PHSS Best Practice Guide
Advice on Best Practice for
CleanroomMonitoring
None in Annex 1
–What system to use?
–Where to locate monitoring points?
–How do we deal with 5µm counts?
–1m3volume 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
CleanroomMonitoring
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 itcurrently 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 Aareas monitored continuously using dedicated particle
counters.
Grade Bareas (background for a Grade A) use dedicated particle
counters.
Other Grade B areas and Grade Careas may be monitored by manifoldsystems to check that they are under control.
(Note: There are no limits for the ‘in operation’state in Grade D areas.)
Corridors and change areasmay be checked on a routine basis using
portable particle countersor monitored using manifold systems.
Enhanced monitoringshould 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
VialSterilizing
TunnelCentralVacuumPumpKeyRemoteCounterVacuumTubing,
Power& DataCentralSoftwareSystem
Appendix B –dedicated counter Grade A
VialSterilizing
TunnelKeyRemoteCounter(Built-in
Pump)
Power& DataVacuumTubingCentralSoftwareSystem0.5µ0.5µ0.5µ0.5µ0.5µ
Appendix B –Manifold for Grade B & C
VialSterilisingTunnelParticleCounterControllerManifoldKeySampleProbeVacuumTubingVialSterilisingTunnelParticleCounterControllerMnifoldKeySampleProbeVacuumTubingVialSterilisingTunnelParticleCounterControllerManifoldKeySampleProbeVacuumTubing
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,
cleanroomclassifications under dynamic conditions,
environmental monitoring, isolators, blow-fill-seal systems
Appendix 1 reiterates FDA’s 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 cleanroomindustry 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”.
CommentsThis is very difficult in practice, and facilities are
normally classified under static conditions as per ISO
14644.Various factors outside the control of
cleanroomcontractors 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 nomicrobiological 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 cleanroomsperforming 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
In operationAt restIn operationStates to be
monitoredCritical = AControlled = C, DGrades A, B, C, DGrade B as
surrounding Grade ARoom
Classes0.5 micron5 micron0.5 micronSizes
monitoredFDA GuidelineEU Annex 1
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 MPPSLeak Testing also at MPPSEfficiency as low as 85% rated as HEPAParticle Counter and CNC only, no photometersA 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
posted by pharma tips @ 1:21 PM
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