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Overview
Integrity testing sterilizing filters is a fundamental requirement of critical process filtration applications
in the pharmaceutical industry. FDA Guidelines require integrity testing of filters used in the
processing of sterile solutions such as large volume paren terals (LVPs) and small volume
parenterals (SVPs). The FDA also requires corresponding testing documentation be included with
batch product records.
Two classifications of integrity testing are destructive and non-destructive. Millipore's practice is to
perform destructive testing as a lot release criteria on samples from each manufacturing lot of all
fabricated sterilizing-grade filter products, and non-destructive testing on each sterilizing-grade filter
prior to sale to insure its integrity.

Destructive Testing
Millipore performs destructive bacterial challenge testing in accordance with ASTM F838-83
methodology. Destructive challenge testing is the best way to determine a sterilizing filter's ability to
retain bacteria. Bacterial challenge testing provides assurance that the membrane and fabricated
device meet the critical performance criteria of a sterilizing filter. The test is performed on a statistical
sample of each lot of membrane and fabricated devices produced.
During the Millipore bacterial retention test, 0.22 m filter discs and devices are challenged with a
solution of culture medium containing bacteria (Brevundimonas diminuta ATCC 19146) at a minimum
challenge of 107 per cm2. The effluent is then passed through a second 0.45 m assay filter disc that
is placed on an agar plate and incubated.

Non-Destructive Testing
Non-destructive testing may be done on filters before and after use. Integrity testing sterilizing filters
before use monitors filter integrity prior to batch processing, preventing use of a non-integral filter.
Integrity testing sterilizing filters after a batch has been filtered can detect if the integrity of the filter
has been compromised during the process. Detecting a failed filter alerts operators to a problem
immediately after batch processing, eliminating delay and allowing rapid reprocessing.
There are three types of non-destructive testing the bubble point test, the diffusion test, and the
waterflow integrity test for hydrophobic filters (HydroCorr Test). The pressure hold, forward flow,
and pressure decay tests are variations of the diffusion test. The stringent requirements of the
pharmaceutical industry dictate that non-destructive filter integrity testing must be performed in each
sterilizing application.
To be able to use an in-process non-destructive integrity test, physical tests were developed that
correlate to the bacterial challenge test. A specification for the physical test correlates directly to the
bacterial challenge test. Once this correlation is established, it is determined that a cartridge passing
the physical test is an integral sterilizing filter.

Bubble Point Test

The most widely used non-destructive integrity test is the bubble point test. Bubble point is based on
the fact that liquid is held in the pores of the filter by surface tension and capillary forces. The

minimum pressure required to force liquid out of the pores is a measure of the pore diameter (see
formula).
The bubble point is expressed as :

Where
k = shape correction factor
= surface tension
= contact angle
d = pore diameter

Bubble Point Procedure

1. Wet the filter with the appropriate fluid, typically water for hydrophilic membranes or an
alcohol/water mixture for hydrophobic membranes.
2. Pressurize the system to about 80% of the expected bubble point pressure which is stated in the
manufacturer's literature.
3. Slowly increase the pressure until rapid continuous bubbling is observed at the outlet.
4. A bubble point value lower than the specification is an indication of one of the following:
- fluid with different surface tension than the recommended test fluid
- integral filter, but wrong pore size
- high temperature
- incompletely wetted membrane
- non-integral membrane or seal

Diffusion Test
At differential gas pressures below the bubble point, gas molecules migrate through the water-filled
pores of a wetted membrane following Fick's Law of Diffusion. The gas diffusional flow rate for a filter
is proportional to the differential pressure and the total surface area of the filter. At a pressure
approximately 80% of the minimum bubble point, the gas which diffuses through the membrane is
measured to determine a filter's integrity. The flow of gas is very low in small area filters, but it is
significant in large area filters. Maximum diffusional flow specifications have been determined for
specific membranes and devices and are used to predict bacterial retention test results.

Diffusion Test Procedure

1. Thoroughly wet the filter with appropriate test fluid, typically water for hydrophilic membranes or
an alcohol/water mixture for hydrophobic membranes.
2. Slowly increase pressure on the upstream side of the filter to the recommended test pressure
provided by the manufacturer, typically at least 80% of the minimum bubble point specification.
3. Allow the system to equilibrate.
4. Measure the gas flow at the outlet for one minute with an inverted graduated cylinder or a flow

meter.
5. A diffusional flow reading higher than the specification is an indication of one of the following:
- wrong pore size
- temperature other than ambient
- incompletely wetted membrane
- non-integral membrane or seal
- liquid/gas combination different than the recommended fluids
- inadequate stabilization time

Diffusion Test

Pressure Hold Test

The Pressure Hold Test, also known as pressure decay or pressure drop test, is a variation of the
diffusion test. In this test, a highly accurate gauge is used to monitor upstream pressure changes
due to gas diffusion through the filter. Because there is no need to measure gas flow downstream of
the filter, any risk to downstream sterility is eliminated.
The pressure hold value is dependent on the diffusional flow and upstream volume. It can be
calculated using the following equation:

Where:
D = Diffusion rate (mL/min)
T = Time (minutes)
Pa = Atmosphere pressure (1 Atm or 14.7 psi)
Vh = Upstream volume of apparatus (mL)
P = Pressure Drop (bar or psi)

Pressure Hold Test

Expert Tips
More

8 Tips to Ensure a Successful Filter

Integrity Test
These tips can help you reduce false failures and improve
robustness and reliability.

Michael Wang, Ph.D.

Click Image To Enlarge +

Filter integrity testing can be conducted manually or via an automated integrity test machine, such as
3M's Minicheck.

A filter integrity test is a critical unit operation commonly employed in the

bioprocessing industry designed to achieve the following:

CONFIRM structural integrity of a sterilizing filter

ENSURE sterilizing filter is properly installed

COMPLY with all regulatory requirements

There are three primary nondestructive integrity test methods that can be used both
before and after filtration:

Forward flow (diffusion) test

Bubble point test

Pressure hold test

Filter integrity testing can be conducted manually or via an automated integrity test
machine. The following eight tips will help reduce false failures and improve
robustness and reliability:

2 TIP #1: Consult Filter Vendor

Product specifications and integrity test methods vary by filter. Seek guidance of
vendor to clarify methods and ensure proper execution of testing.

3 TIP #2: Select the Proper Integrity Test Method I: Hydrophilic vs.
Hydrophobic
Click Image To Enlarge +

Figure 1. Forward flow integrity test (manual method)

4 There are two types of sterilizing membrane filters based upon chemical
properties: hydrophilic and hydrophobic. For hydrophobic sterilizing
membrane filters, a water intrusion test (a secondary, nondestructive integrity
test method) is recommended if the primary, nondestructive integrity test
methods prove problematic.

5 TIP #3: Select the Proper Integrity Test Method II: Membrane
Structure, Filter Area, & Pore Size

Bubble Point Test: best suited for testing symmetric membrane filters
and small filter installations (e.g., 47 mm disc or single capsule).

Forward Flow Test (Figure 1): best suited for testing asymmetric
membrane filters and large filter installations (e.g., multiple cartridges or capsules).

Note that integrity test methods are only valid if they can be correlated with bacterial
removal efficiency. Thus, forward flow, bubble point and pressure hold integrity
test methods are suitable for sterilizing membrane filters with pore sizes ranging
from 0.100.45 micron.

6 TIP #4: Select the Proper Integrity Test Method III: Manual vs.
Automated Integrity Test
Manual Integrity Test:

Pro: easy to setup and execute.

Con: test tubing/apparatus connected to the downstream side of the

filter (risk of contamination significant).
Automated Integrity Test:

Pros: instrument connected to upstream side of filter (risk of

contamination low); hard record of data (via computer or printout); elimination of
operator subjectivity.
Based on pros/cons of both test methods, the automated integrity test is preferred
whenever possible.

7 TIP #5: Maintain Temperature Control (Automated Integrity Test

Method)
When using the automated integrity test method, ensure the temperature remains at
ambient throughout the entire process. Any fluctuation in test temperature can
trigger significant change in pressure and/or volume, resulting in a false failure.

8 TIP #6: Eliminate Sources of Surface Tension

Thoroughly rinse/flush the empty test housing and all related components (e.g., orings). It is imperative that all components utilized are free of surface tension
suppression agents (e.g., oil from a new o-ring), as their presence can generate
false failures.

9 TIP #7: Ensure the Test Housing is Properly Sealed & Vented
When installing a filter in the test housing, take great care to ensure filter o-rings
dont become dislodged from respective grooves. Similarly, ensure the housing
sump is seated properly on the base o-ring. Proper venting is achieved when a
steady stream of liquid (i.e., no air) emits from the vent.

10 TIP #8: Adhere to a Robust Membrane Wetting Protocol

Finally, the most common mistake that causes false integrity test failures is an
inadequately saturated (wet) filter. 3M Purification recommends one of
the following methods to ensure a completely wet filter prior to conducting an
integrity test:
Soak: stationary exposure of the filter to be tested in the appropriate

wetting solution for a minimum of 10 minutes.

Flush: dynamic exposure of the filter to be tested in the appropriate

wetting solution for approximately 5 minutes @ 3 GPM (11.4 LPM) per 10 filter.
Michael Wang, Ph.D., is Senior SASS Specialist II, Scientific Applications Support Services at 3M
Purification.

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Filter Integrity Testing in the

Pharmaceutical Process Environment
Automating filter integrity testing helps to ensure that GMP
sterility requirements are met.
Jun 01, 2006
By Reinhard Baumfalk, Jerry Finazzo
BioPharm International
Volume 19, Issue 6

Integrating filtration steps into an aseptic

pharmaceutical process has some unusual
complications. Various factors, such as particle load,
fluctuations in pressure and temperature, and cleaning
steps (which may have high temperatures or severe
Reinhard
changes in pH), can cause wear to the filter elements. Baumfalk, Ph.D
Continued performance of the filter elements becomes
one of the critical process parameters that must be
controlled.1,2,3 Using a filter in a process can lead to various damage
phenomena such as irreparable blockage of the filter membrane and
cracks or changes to the membrane or pore structure. While blockage
can be easily detected during the process run, cracks or changes in
pore structure cannot be detected that easily.4
There is a method for checking these filter
element properties throughout the course of
the processthe filter integrity test. A typical
four-step integrity test procedure (algorithm)
is Figure 1.5 We will delve first into the test
methods and then we will describe how to
ease the burden by automating the entire
cycle. Many industry-specific terms are
defined in the glossary at the end of this article.
INTEGRITY TEST METHOD
The four integrity test steps are implemented upstream and
downstream of the filtration unit in order to be able to detect any
possible changes in filter element properties and thereby supply
positive results to confirm the success of the filtration step. The
diffusion test and the bubble point test are established, nondestructive test methods. To determine validity, compare the test
results to the filter manufacturer's tolerance data. The bases of the
tolerance data are validation processes conducted by the
manufacturer. These are used to determine tolerances by correlating

absolute, destructive test methods (bacteria challenge test) with filter

integrity tests. Both test methods rely on a procedure of wetting the
filter membrane with a pre-defined medium (typically ultrapure water,
but also customer-specific liquids).4
To start the automatic filter test, the filtration housing is disconnected
from the actual process, or the filter is installed in a separate test
housing. In this procedure, the filter element is pressurized at a specific
value on the upstream (non-sterile) side. At this test pressure, the
diffusion through the filter membrane is determined for values starting
at 0.1 mL/min. By contrast, when the bubble point test is carried out,
the applied test pressure is raised successively and the diffusion
measured in parallel is monitored to determine any disproportionate
increase. This increase in diffusion is caused by the wetting medium
being pressed out of the pores of the filter membrane, which initially
occurs in the largest pores, thereby providing information about the
upper limit of the pore size distribution. Typical bubble point values for
microfilters range between 3,000 mbar and 4,000 mbar and, in
ultrafiltration, can go up as high as 8,000 mbar. These methods also
allow reliable detection of cracks or blockage.
Air filtration also needs a filter integrity test method to guarantee the
sterility of critical parameters. Applications for these filter elements
include pharmaceutical and biotechnological production systems.
Unlike liquid filtration, air filtration elements are based on hydrophobic
membranes, which prohibit the test methods previously mentioned
(membrane wetting with ultrapure water).
In this context, an alternate, non-destructive methodthe water
intrusion test (WIT)6 is an established test. During the WIT test, or the
very similar water flow test (WFT), the penetration of water into the
hydrophobic membrane is measured under pressure and evaluated by
comparing the results to a limit value. Additionally, this filter type can
be tested by a diffusion test after wetting the membrane by liquids
with low surface tension such as isopropanol or mixtures of WFI and

isopropanol.

Air filters are sterile barriers that typically are sterilized prior to the test
step (Process step 1 in Figure 1), at the same time the production
system is sterilized. They can no longer be removed for the test or
during the test, but may only be accessed from the non-sterile process
direction. Since the sensitive measurands (diffusion, bubble point or
water intrusion value) can already be determined within the upstream
volume, no contact or inflow to the sterile area is necessary and
process-relevant sterile barriers remain
intact.
All of these test methods give results as
either a flowrate (diffusion test, water
intrusion test, water flow test) or a pressure
value (bubble point test, pressure drop test)
which has to be compared to the given limit
value of the tested filter element in order to Table 1. Typical limit
pass the test. These limit values are supplied values for different
either by the filter manufacturer as part of
filter types
the validation documents of the filter
element, or are determined during a validation process for customerspecific parameters and correlate to a destructive bacteria challenge
test.7 Table 1 shows a typical set. Short test descriptions appear in the
glossary.
AUTOMATED FILTER TESTING
The test algorithm described by Figure 1 consists of several steps to be
performed by an automated integrity tester (Figure 2). An automated
integrity tester therefore needs to be able to handle pressurized air or
WFI, in order to pressurize the filter cartridge with the appropriate
media. Based on this requirement, hardware
such as valves, pressure transducers, and
flowmeters are standard components of a
Figure 2. Standard
today's integrity tester. Typically these parts integrity test algorithm
are controlled by micro-controller-based
11
hardware and run by proprietary firmware
(GAMP class 2 or 3).8Since integrity testers are not only used in a welldefined laboratory environment, but also installed in a rough
production process, the integrity tester manufacturer has taken these
conditions into consideration. Hardware components should be made

of appropriate materials (stainless steels are favored) and have the

needed process reliability. Software has to cover the normal operating
conditions, and also be able to detect critical changes of process
parameters such as temperature or supply media (pressurized air).
Often, these requirements are met by an automated functionality test
or separate software control loops within the test procedure.
To meet the needs of process integration, most of the available
integrity testers have interfaces in order to use additional external
hardware. External valves mounted on the filter housing can be
handled directly, or controlled by sending commands via a PLC
interface. External sensors can be connected in order to perform the
measurement directly at the filter housing, because long
tubing lengths could interfere with the accuracy of the
measurement.
Because the WIT and WFT test use WFI as penetration
media for the filter membranes, additional dedicated
equipment has been developed by most of the integrity
test manufacturers. A trolley equipped with a stainless
steel vessel stores the needed water volume under
Filter
sterile conditions. A PLC allows automated CIP or even
Integrity
SIP procedures for the vessel, with valves permanently
connected.

Electrical interfaces are specified based on serial protocols, such as

RS232 or RS485, or based on the Ethernet/TCP/IP protocol. These
interfaces allow a data transfer, or even a remote control of the
integrity tester, by a PLC or MES software.
PROCESS INTEGRATION
Data handling involved in integrating an integrity tester into a filtration
process can generally be accomplished in one of two ways: electronic
data storage in a 21 CFR Part 11 environment or hard copy printouts

where no data are stored. Hard copy is not covered under

21 CFR Part 11.9,10
In addition to deciding on the method of data storage,
the integrity tester needs to be qualified before it can be
used in the production environment. Qualification is
typically divided into three steps, namely Installation
Qualification (IQ), Operational Qualification (OQ) and a
Performance Qualification (PQ). We cover this later.
When working with electronic data storage, many factors Glossary
must be considered to properly integrate the integrity tester into the IT
process. Not only must the integrity tester fit into the 21 CFR Part 11
environment, but also all other equipment (PLCs, computers, SCADA
systems) involved in the data transfer must fit. Also, one must consider
the method of data transfer. There are many standards in industry for
equipmentCOMMUNICATION . Some of the more common ones found
on integrity testers are Ethernet and OPC servers for communication to
computers and RS-232 and Profibus DP for communication to PLCs.
Pay attention to when and how often data are to be transferred from
the integrity tester. Integrity testers usually have the ability to transfer
data automatically (as it is being created or time based) and manually.
It is important that the data be transferred in a timely fashion to a
designated computer where the data can be properly archived and
backed up. While integrity testers may have the memory to store large
amounts of data, they are not intended as the sole data storage
location for integrity test data (transient data).
In principle, two strategies for electronic data handling are common.
First a strategy described as the "stand alone" setup is based on the
capability of the state-of-the-art devices to process and store data. The
device is only connected from time to time to the process network
containing the archive servers or MES systems. This device needs to
have the capability to store data (integrity test parameters as well as
test results) since GMP-relevant data are being stored. An audit trail, or
user access management, is required since all 21CFR Part 11
requirements have to be met in the device.
In the second setup, the devices are permanently connected to the
process network. Due to this permanent connection, in most cases all

relevant GMP data are not stored on the device, but directly received
from or sent to the MES or PLC system ("unique source principle"). The
supervising network can handle the start of integrity tests and running
the complete test procedure. Since GMP data are not stored on the
integrity test device, it is only a part of a 21 CFR Part 11 system.
If electronic data files of test results are stored on the integrity tester,
the 21CFR Part 11 requirements have to be fulfilled as mentioned
above. But because the software capabilities of such a system are
much less than that of a common PC, smart solutions have to be
found. For instance, electronic data should be stored in separate files.
By adding a checksum to each file and naming it with a unique nonreproducible name, it will have the required 21 CFR Part 11 features.
Then, the computer or data historian (audit trail) can link the data file
to the proper filter and product batch records. This is commonly
described as traceability and the file content can be checked for
integrity based on the specific checksum. The data file includes all test
parameters as well as the actual integrity test value with a definitive
pass or fail result. Typically, additional information is also included in
the integrity test result file such as product, product lot, filter part
number, and filter lot or serial number.
A large majority of existing processes do not have electronic data
transfer capability and are organized by a paper-based documentation
system. Most integrity testers on theMARKET will have an onboard
printer, or a serial port for an external printer. A printout is the only
record of the integrity test and it is essential to make a photocopy.
Typically, one copy is stored in a safe location while another is
incorporated into the batch records for each process run.
QUALIFICATION
Qualification of an integrity tester is an important part of the process
integration. Suppliers of integrity testers support customers with the
device, and with qualification protocols that are optimized for an
efficient and secure qualification process based on the supplier's knowhow. Qualification support is typically available from the supplier and
ensures that the qualification can be performed in a professional
manner. The documentation used, whether the integrity test results

are electronic records or hardcopy printouts, is very similar. Additional

protocols are used when data transfer is involved.
Installation qualification. A qualification generally starts with the IQ.
The purpose of the IQ is to provide documented verification that the
integrity tester complies with approved design intentions and the
manufacturer's recommendations or requirements. Verification that the
proper documentation has been supplied with the unit, a physical
inspection of the equipment, and a facility requirements check are
common items included in the IQ.
Operational qualification. The purpose of an OQ is to provide
documented verification that the equipment performs as intended.
There are many items verified during an OQ, with some of the more
difficult tests confirming that the actual integrity test result is within
specification. Typically, for bubble point, diffusion (forward flow), and
water intrusion, the integrity test result will be compared with a
measured reference. For example, when testing for diffusion, it is
common for the integrity result to be verified using a mass-flow meter.
IQ and OQ can be performed by the manufacturer of the integrity
tester, or by the end user. Similarly, the IQ and OQ protocols will
typically originate from the manufacturer, with the end user using
them as is, or modifying them to their validation standards.
Performance qualification. The last part of a validation is the PQ.
The purpose of a PQ is to provide documented verification that the
equipment can perform effectively and reproducibly based on
approved standard operating procedures (SOPs). Typically, actual
process filters in the production environment are used during this
testing. The end user's SOPs, as well as
personnel, are used to perform the PQ. The
integrity tester manufacturer may be consulted
during the PQ, but typically will not perform the
protocols.
CONCLUSION
The integration of filter integrity testers impacts
the filtration process, as well as the IT
environment. Available integrity testers offer a

Quick Recap

wide variety of integration options to perform in a laboratory and also

in the process environment. Because the integration concept is based
on a standard device, which only has to be configured in an
appropriate way, qualifying the device within the customer's validation
process can be done by using the integrity tester manufacturer's
documentation. This possibility leads to dramatically reduced efforts in
the qualification procedure and should therefore result in a preferred
choice of standard integrity testers to carry out a filter integrity test
step in a filtration process.