In t r o d u c t i o n t o

In t e g r it y T e s t in g
Theor y
G. Somasundaram (Som)
Associate Director Technology Management
South East Asia & Oceania
Singapore

Somasundaram.g@merckgroup.com
16th May 2017

Merck KGaA
Darmstadt, Germany
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Agenda

1
Why and When to Integrity Test

2
Regulatory Requirements

3
Introduction to Integrity Testing

4
Considerations for a Reliable Integrity Test
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Sterilizing Grade Filter?

•Functional definition
• Remove microorganisms from a fluid
• Produce sterile fluid

•Limited value in defining as a pore size

•Define in terms of Bacterial Retention

• ASTM method
• Retains >107 cfu B. diminuta per cm2 of filtration area
• Produce sterile filtrate

•Microorganisms limited to bacteria, yeast, molds, protozoa

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Basic Elements of a Bacterial Retention Test

ASTM F838 05
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What is Membrane Integrity?

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Why perform Integrity Testing (IT)?

• Check correct installation

 Detects system leaks due to o-rings,
gaskets, faulty seals

• Confirms manufacturers specifications

 Assures the correct pore size filter

• Check for damages

 Assures integrity before sterilization
 Assures integrity after steaming or
autoclaving

• Regulatory requirements
 Link between validation and current
processing conditions
 GMP/Audit Requirement
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Where & When Do We Integrity Test?

WHAT DOES IT FIND? WHEN CAN YOU DO IT?

Faulty housings Before sterilization

Out-of-the box failures

Sterilization induced failures Before use

Stress (process) induced failures After use

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Which Filters to Test & When

Biotech Manufacturing Process

Should all sterilizing grade filters (0.2um) used in the process be tested?
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Which Filters to Test & When

Biotech Manufacturing Process

For final
product
sterilizing
filters:
• Post-use IT:
Regulatory
requirement
• Pre-use IT:
required in EU
and otherwise
recommended
to consider as
part of risk
assessment

Perform IT based on Regulatory

Risk Analysis Requirement
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Who defines the requirements?

Among others!
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Regulatory Requirements

EU GMP: The integrity of the sterilised filter should be verified

before use and should be confirmed immediately after use by an
appropriate method such as a bubble point, diffusive flow or
pressure hold test
EU GMP Annex 1 for the manufacture of
sterile medicinal products, February 2008

US FDA: Integrity testing of the filter can be

performed prior to process and should be routinely
performed post-use.
US FDA Guideline on Sterile Drug Products Produced
by Aseptic Processing, 2004

SFDA: After sterilizing filter is used, the integrity of

the filter should be tested and the results recorded.
The test methods could be bubble point, diffusion or
pressure hold.
SFDA Guidelines, 2010

Where sterility is claimed: IT before use

is recommended, after use is required
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Industry Guidance

PDA® TR26:
• Where the claimed purpose of the filter is to sterilize, pre and post
filtration integrity tests should be performed.
• Steps should be taken to ensure that the downstream side of the
system remains sterile when performing a post sterilization, pre-
use integrity test.
PDA® Technical Report 26-2008, pg. 34

ISO® 13408: The filtration system should be

designed to permit in-place integrity testing as a
closed system prior to filtration.
ISO® 13408, 2003

PDA® recommends IT pre-use and post-use.

Industry guidance emphasizes that maintaining
process sterility is of critical concern
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General Types of Integrity Tests

Non-Destructive: Capillary or diffusive by Manufacturers for process testing

Capillary
− Bubble point, water intrusion
− Direct correlation to pore size/retention
Diffusion Based
− Diffusion, HydroCorr, Forward Flow (same as Diffusion)
− Predictive of bacterial retention
− Sensitive check for bulk flow

Destructive
Bacterial Retention
− Actual demonstration that the filter will perform designed task
− Destructive and impractical for in-situ
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Common/Regulatory accepted types of In-process Integrity Tests

Hydrophilic Filters (Liquid) Hydrophobic filters (Gas & vent)

− Bubble point (BP) − HydroCorr (Water Intrusion)

− Diffusion (Forward Flow) − Alcohol BP

− Enhanced (Diffusion + BP) − Alcohol Diffusion

“A physical integrity test is meaningful only when it can be related to specific

filter retention characteristics.” PDA® Technical Report 26 Section 7.2

BP, Diffusion and Water Intrusion are correlated to Bacterial Retention

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Destructive vs. Non-destructive Testing

There must be a correlation between a

destructive and a non-destructive
integrity test
 Regulatory requirement
 FDA Aseptic Guidelines
 “A physical integrity test is meaningful only when it can be related to
specific filter retention characteristics.” PDA® Technical Report 26
 Validation justification for the use of a test

Where,
LRV=log10 [# of organisms in challenge solution/# of organisms in filter effluent]

For Example,
1X108cfu in challenge and 0 organisms in effluent would be reported as LRV>/= 8
“>” symbol means 0 in denominator of above equation
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How Capillary forces influence IT?

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Manual Bubble Point Testing

Flush with water Apply air pressure

Increase pressure until rapid continuous

flow of bubbles is seen downstream
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Setup Checks - Automatic

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Physical integrity test – Diffusion

Wet
Membrane
Gas is dissolved in liquid
Apply
40psi pressure from is held in the pores of a
validated fully wetted membrane
Upstream
High Pressure
specification filter.
Hold
Pressure A pressure differential
will give a gas
concentration gradient
across the filter.
Results in diffusive
Downstream
gas flow from
Low Pressure upstream to
downstream.
Physical integrity test – Diffusion
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Out of specification

Gas Flow
40psi
Hold Good
Upstream
High
Pressure Bad Filter Filter
Pressure

Within
specification

Pressure

Downstrea For this test, you are measuring

m Low that gas flow at the prescribed test
Pressure
pressure meets validated
specifications.
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Physical integrity test – Bubble Point

30psi 40psi ≥50psi Gradually
Bubble point is the transition point
raise
pressure from diffusive to bulk flow, where
liquid gets evacuated from the
Upstream Wet
largest pores.
High Membrane
Pressure
Water held in For this test, you are seeking to
by capillary identify this transition point which
forces must align with validated filter
Sufficient pressure will specifications.
over come the capillary
forces to evacuate the
liquid from the filter.
Downstrea
m Low
Pressure
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Physical integrity test – Bubble Point

Bubble point is the transition point from
diffusive to bulk flow, where liquid gets
evacuated from the largest pores.

For this test, you are seeking to

Upstream
High identify this transition point which
Pressure must align with validated filter
specifications.

Out of Within

Gas Flow
specification specification
Downstrea
m Low Bad Filter Good Filter
Pressure

Durapore
® CVGL:

50psid

Pressure
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Hydrocorr/Water Intrusion Test for Hydrophobic Filters

Install dry  Hydrophobic polymers repel water

 Water Intrusion Pressure is
2.6

cartridge
the minimum pressure necessary to
bar
0
Fill force water into the largest pores of
upstream a hydrophobic (water resistant)
w/ water membrane.

Apply
pressure
Hold
pressure

Pressure causes pleats

P atm
P atm
to compact - resulting
in apparent water flow

Measure pressure
decay
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Hydrocorr/Water Intrusion Test for Hydrophobic Filters

 Hydrophobic polymers repel water

 Water Intrusion Pressure is
2.6

the minimum pressure necessary to

bar
0
force water into the largest pores of
a hydrophobic (water resistant)
membrane.
Water Out of
Flow Bad Filter Specification
ml/min

1.0 Aervent®:
0.75 ml/min for
10”
0.5
P atm
P atm
0.4 Good Filter
0.3 20 Within
minutes Specification
0.2
0.1
Measure pressure
decay 0 10 Time
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What can Affect the Integrity Test

Bubble Point Diffusion

 Wetting along and through the device  Wetting along and through the device
 Pore shape  Wetting solution
 Wetting solution  What is on the pore
 What is on the pore  Test gas
 Pore diameter  Temperature
 Temperature  Differential pressure
 Porosity
 Area
 Wetted thickness

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What is the Best Integrity Test?

The one that

 Gives the greatest sensitivity/reliability
 Gives the best accuracy
 Can be done in the test environment
 Uses specifications provided by the manufacturer
 Can be done with the test equipment available
 Fits in with the company philosophy

Sometimes it is diffusion, sometimes it is bubble point, sometimes is it both

All non destructive integrity tests measure or detect gas flow

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Manual vs Automated Integrity Testing

Manual Automated

Advantages
Advantages  No downstream interventions
 Simple equipment  Facilitates post-SIP testing without risking
 Direct measurement or downstream contamination
observation of gas flow  Repeatable, can be validated
 Fast  Eliminates Operator subjectivity
 Enhanced record keeping
 Hard Copy Printout enabling
Disadvantages Troubleshooting
 Nearly impossible to perform
aseptically Disadvantages
 Operator Subjectivity  False Failures
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Co n s i d e r a t i o n s f o r a
R e lia b le In t e g r it y T e s t
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Considerations for a Reliable Integrity Test

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Test Selection

Hydrophilic Filters Hydrophobic filters

(Liquid) (Gas & vent)
 Bubble point (BP)  HydroCorr (Water Intrusion)

 Diffusion (Forward Flow)  Alcohol BP

 Enhanced (Diffusion + BP)  Alcohol Diffusion

Test
Selection What needs to be
FTET considered in selecting
the right test?
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Selecting the right test depends on the properties of

the filter and your process

Filter
Characteristics

Hydrophobic or Pore size

Membrane area Pore structure
hydrophilic >0.45 to 0.1µm

Process
Characteristics
Test method Environment
•Manual v. •Temperature
Automated stability
Test
Selection

FTET
Your supplier should help to recommend the
best test for your process.
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Certified Operator: Critical Link to Sterility assurance

Program information
Operator and lot info
Test Specifications
Upstream volume
Diffusion
Bubble point
Summary

FTET
Trained Operators trained to interpret the test results is
Operato
r
critical to verify the validity of the integrity test.
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Robust Wetting Procedurec

• BP and Diffusion require a fully wet filter

• Bulk gas flow through dry pores results in false failures
• Most common reason for improper wetting is flow rate and pressure are
not defined or controlled
• Robust wetting procedure design should be based on -
• Manufacturers
recommendation
• Understanding
membrane
wetting
characteristics 1 LPM/ft2 (0.1 m2) filter area for 5
• Process capability, minutes
control and
monitoring

Wetting Instructions – P35515

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Stable Temperature Environment

• Manufacturers Specifications are typically

established at ambient temperature (~23°C)

• If operating outside of ambient temperature, a

ratio should be established during validation and
applied to test specification
• E.g.: Cold room operation or heat jacketed
housing Effect of Temperature on Diffusional Flowrate

Correction Factor (Relative

2.2
2.0
1.8

to 23 C)
1.6
1.4
1.2
1.0
0.8
5 10 15 20 25 30 35 40 45 50 55 60 65 70
FTET Temperature (C)
Stable
Environment
There is no significant impact on diffusive flow or BP
over normal ambient temperature range
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Stable Temperature Environment

When using Automatic Testers, temperature changes during a test
leads to additional upstream pressure gain or loss – Test Error
 Ideal Gas Law - PV = nRT
 As Temperature increases Pressure increases
 As Temperature decreases Pressure decreases
Trained Operator can identify the Temperature effect by reviewing the
flow curve
Even a change of 1oC DURING the test can lead to false failure because
•
the pressure change WILL NOT be solely due to gas flow
Air Flow Diffusion flow Out of
Common reasons for ml/min Specification
temperature changes
Flow at Unstable Temp
• Inadequate stabilization False Failure
After steaming or wetting with hot
WFI
15
Durapore®:
• Environmental changes 13.3 ml/min for
during testing Near autoclave, 10 10”
HVAC outlets, Cold room door, Near Flow at Stable Temp
a window
5 Within
Specification

1
FTET
Stable 10
Environment
0
Test has to be repeated per retest Time min

SOP after establishing stable test

conditions
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Validated Equipment

Regulatory agencies require instrument

validation to ensure reliable and
Design Specification predictable performance
and Qualification
Establish and provide documentary evidence

Change Control
that:
Installation
Qualification  DQ: Instrument is designed
Change Control in accordance
with the requirements of GMP. (Supplier)
Changes to instrument initiates the
Operational IQ: Instrument
design process athas• been
the URS built and
supplier.
Qualification Itinstalled in compliance
should follow with their
Functional
the• same design
Specification
qualification
specifications.
process (Supplier)
steps – DQ, • Design Specification
IQ, OQ, PQ.
• Component Construction
Process Validation  OQ: The supporting utilities and the
• Acceptance Test
instrument operate in accordance with
• Module Test
their design specifications. (Supplier)
• Integration Tests
 PQ: A specific process
Regulatory will consistently
requirements
• Systems are
Acceptance Test
FTET produce a product
established • meeting
via DQ, IQ,itsOQ
Validation Summary
Validated predetermined specifications
and PQ. Suppliers and quality
typically
Equipment
attributes. (End User
offer protocols and with Supplier
execution
Assistance)
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Fluid properties affecting IT results

• Feed stream components can impact

surface tension and/or gas diffusion rate
impacting IT results as compared to
suppliers ( typically water based)
specifications
• This must be controlled or validated.

Common causes for alteration:

• Contaminants: New silicone tubing, residual
cleaning agent, etc
• Identify and eliminate
• Residual Product: Develop product
based integrity test specification or post
use flush procedure

Consistent Application
Test Fluids Note
FTET AN1505EN00
During filter validation, your supplier can help to
establish product specific integrity test
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Troubleshooting SOP with Decision tree

SOP should
• Be based IT physical
principles and ability to
interpret test results
• Minimize re-tests & improve
overall test efficiency
• Link to the validated process
parameters
SOP should address the following
• Is the filter integral?
• If it is a false failure, why did it
fail?
• Leaks or temperature changes
• Poor wetting PDA® Journal of Pharmaceutical
• Surface tension suppression Science and Technology, Vol. 62, No.
S-5, 2008
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Pr o d u c t
S pe c if ic
In t e g r it y
T e s t in g
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Product-Wetted versus
Water-Wetted Integrity Testing

• Filter manufacturers recommended test values are for specified fluid(s) and conditions.

• Testing with non-specified fluid(s) conditions requires establishing product/process specific

test parameters

• Testing includes examining the effect of fluid AND process variables on integrity test results

and the reproducibility of these values

PDA Tech Report 26 Sec. 7.3

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Product Specific Test Principles

Compare water to product on the same filters

Variables that could effect bubble point include:
 surface tension
 contact time/volume
 membrane lots
 temperature
Bubble point should test multiple filter lots
Diffusion can be multiple trials on one filter
Product based specifications should be based on experimental results, rather than calculations

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Summary

• IT is a critical component of Sterility

Assurance and is a regulatory
requirement

• The choice of an integrity test depends

on the size and type of the filter and
your risk mitigation strategy.

• Good process design will deliver

reliable IT operation.

• Validated, automatic integrity testers

improve process efficiency.

• We are here to help.

T h a n k Yo u

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