The document discusses MilliporeSigma's helium integrity test technology for validating the integrity of single-use systems. It provides an overview of regulatory guidance on integrity testing, outlines MilliporeSigma's three pillar strategy for integrity assurance, and describes how the helium integrity test works and its ability to detect defects down to 2 microns in size. Mobius single-use assemblies are tested using this method and receive a certificate of quality to verify 100% integrity testing.

1. The life science business of Merck KGaA,
Darmstadt, Germany operates as
MilliporeSigma in the U.S. and Canada.
How a Helium Manufacturing
Release Test Increases
Integrity Assurance for
Single-use systems
March 31st 2022

2. The life science business
of Merck KGaA, Darmstadt,
Germany operates as
MilliporeSigma in the U.S.
and Canada

3. Agenda
1
2
3
Regulatory & Industry
Guidance
Life Cycle Management /
Risk Mitigation
Correlation to Microbial
Ingress
4
5
Helium Integrity Test
Technology
What to Expect-Mobius®
Single-Use Assemblies
tested with Helium
Integrity Test

4. Regulatory &
Industry
Guidance

5. Helium Integrity Test Technology | 31.March.2022
5
Industry Guidance & Standards
Guidance Standard
BPSA
(Bio-Process Systems Alliance)
Design, Control, and Monitoring of Single-Use Systems for Integrity
Assurance.
PDA
(Parenteral Drug Association)
Technical Report 27 Pharmaceutical Package Integrity.
ASTM
(American Society for Testing and
Materials)
ASTM F2391C05 Standard Test Method for measuring Package and Seal
Integrity using Helium as the Tracer Gas.
USP
(United States Pharmacopoeia)
USP <1207> Sterile Product Packaging Container Closure Integrity
Evaluation.
EU GMP
(European Commission)
Annex 1 Manufacture of Sterile Products, draft, 2020
ISO
(International Organization for
Standardization)
ISO 11607-1: Packaging for terminally sterilized medical devices
ISO 15747: Plastic containers for intravenous injections

6. Annex 1 Draft. Manufacture of Sterile Medicinal Products
Regulatory Changes
Helium Integrity Test Technology | 31.March.2022
6
Draft Guidance, not yet published.
8.119 Appropriate measures should be in place to ensure the integrity of components used in aseptic
connections. The means by which this is achieved should be determined and captured in the CCS*.
Appropriate system integrity tests should be considered when there is a risk of compromising
product sterility. Supplier assessment should include the collation of data in relation to potential failure
modes that may lead to a loss of system sterility.
8.121 SUS are those technologies used in the manufacture of sterile products [...].
8.112 There are some specific risks associated with SUS which should be assessed as part of the CCS*.
These risks include but are not limited to:
i. [....]
vi. The risk of holes and leakage
vii. The potential for compromising the system at the point of opening the outer packaging.
* CCS Contamination Control Strategy

7. Regulatory & Industry Guidance
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7
USP <1207> : Container
Closure Integrity Testing
No regulatory guidance
on single-use system
integrity
Regulations for
Container Closure
Integrity (CCI) of
primary drug
packaging
Customers & industry
groups apply CCI
requirements to single-
use systems
BPSA
ISO
ASTM
A Quality risk-management approach is recommended

8. Life Cycle
Management &
Risk mitigation

9. ✓ Integrity Assurance – a holistic approach of risk analysis and mitigation
by means of product and process robustness, quality and process control
and integrity testing
✓ Leak Test – a test used to identify leaks of certain sizes in a single-use
system
✓ Integrity Test – a test used to confirm the defined barrier properties of a
single-use system
✓ Non-Destructive Test Method – a test method that maintains the tested
single-use system in a conditions for further use, without impacting its
quality attributes
✓ Destructive Test Method – a test method that may destroy the tested
single-use system during the test and prevent from further use
✓ Maximum Allowable Leakage Limit (MALL) – the greatest leakage rate
(or leak size) tolerable for a given product package that poses no risk to
product safety and no or inconsequential impact on product quality
Terminology
Lets start on good grounds
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9

10. Single-Use System Life Cycle
Leak risks should be mitigated at every step
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Assembly
Manufacture
Assembly
Design
Release
Testing Pre-Use
Integrity
Testing
Receipt &
Unpackaging
Post-Use
Testing
Sterilization &
Shipping
Manufacturer End-User
Critical point in
the lifecycle!
Last step that the
manufacturer has
direct control over

11. Business Integrity
Operator
Environment
Product
Risk Management Plan
Four Main Elements
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11
Risk to Patient
• Quality by Design
• Quality Risk
Management
• Handling &
Operator Training

12. Three Pillar Integrity Assurance Strategy
12
Three Pillar
Integrity Assurance
Strategy
Quality by Design
(QbD)
Handling & Operator
Training
Quality Risk
Management
(QRM)
▪ Initial component
qualification &
specific testing
▪ Junction Validation
▪ Engineering design
assessment
▪ Best Handling
Practices (Bags &
Assemblies)
▪ Operator Training
▪ Single Use Visual
Acceptance Criteria
Library
▪ In-Process Testing
▪ Release Testing
▪ Packaging &
Transport Validation
▪ Point of use
Integrity Test
Helium Integrity Test Technology | 31.March.2022

13. Leak Test Integrity Test Helium Integrity Test
Leak Test vs Integrity Test
Release Test
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Integrity Test
Detects gross leaks in a system,
such as:
 Improper bonding of film
 Missing components
 Improper tubing connections
 Deficiencies in bag making and system
assembly
Detects defects of a known size that have a correlation to a known
parameter using a specific test methodology such as aerosolized or
liquid bacterial ingress.
 Offers a high level of confidence in the integrity of the single-use system
 Recommended for critical applications such as final fill single-use systems
 Needs to be validated and correlated to microbial ingress
Leak Test
Decreasing Defect Detection Size Limit
Increasing Level of Risk-Reduction

14. Test Sensitivity
Release Test
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Leak Test Integrity Test Helium Integrity Test
Integrity Test
Leak Test
System Volume
Approximate
Detection Limit
Tubing Assemblies 50 µm
1 – 9 L 150 µm
10 – 999 L 1000 µm
>1,000 L 2000 µm
System Volume
Validated
Detection Limit
Tubing Assemblies 20 µm
Up to 50 L 20 µm
System Volume
Validated
Detection Limit
Tubing Assemblies 2 µm
Up to 50L 2 µm
Select 2D 50 L Single Use Systems can obtain 50 to 500 times better sensitivity

15. Not a “one-size-fits-all” Approach
Integrity Test Technologies
Test Feature Tracer-gas Integrity Test Air/Pressure Integrity Test
Sensitivity ≥2 µm ≥10 µm
Impact of Test Environment
Impact of Test Specimen Volume
Impact of Test Specimen Materials
SU Assembly Design Space
Test Time
Maintenance
Investment Costs
Low/Simple
Medium/Challenging
High/Complex

16. Test Complete Single-Use Systems
Design Space Considerations
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Single Use Component Included
Bags
Tubing
Other components*
Connection Points
* Depend on design, some
components cannot be test.

17. 2D 50 L Single-Use System
Design Space Considerations
17
• Filters up to 30”
• Multiple tubing types,
diameters and lengths
• Limitation on total single
use system volume
Helium Integrity Test Technology | 31.March.2022
There are design space limitations, not every
system can be tested via Helium Integrity Test

18. Correlation to
microbial
Ingress

19. Two Test Methods
BPSA Guidance on Critical Defect Size
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Aerosol Challenge Liquid Immersion Challenge
19
2017 Bio-Process Systems Alliance Design, Control and Monitoring of Single Use Systems for Integrity Assurance

20. Results
Aerosol And Liquid Challenge
Helium Integrity Test Technology
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2017 Bio-Process Systems Alliance Design, Control and Monitoring of Single Use Systems for Integrity Assurance

21. Helium Integrity
Test Validation

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In-House Verification of 2 µm Critical Defect Size
Helium Integrity Test Validation
Test Parameters
Test Bags 2L 2D bag
Growth Media
Trypticase soy
broth
Challenge Organism
Brevundimonas
diminuta
Microbial
Concentr
ation
T= 0 hrs 2.07E+07
T = 3 hrs 2.20E+07
Immersion Time 3 hours
Incubation Temp 36°C
Incubation Time 2 weeks
Defect Size (µm)
Results
Positive/Total
2 0/6
5 3/3
10 2/6
15 4/5
20 3/3
30 5/5
40 5/5
50 4/4

23. Objective:
 Screen the compatibility of individual
components from the Mobius
component library
 Determine the systems acceptance
cut-offs
Procedure:
 Permeation characterization of 2D bags
 Testing of tubing and other components
 Min/max assembly volume testing
Helium Integrity Test Validation
Validation of Equipment
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0.00E+00
2.00E-07
4.00E-07
6.00E-07
8.00E-07
1.00E-06
1.20E-06
No Defect Defect
Leak
Rate
(atm*cc/sec) 5L 10L 20L 50L
2D Bag Leak Rates
(No Defect vs 2 µm Defect)

24. Helium
Integrity Test
Technology

25. 25
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P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
Industry
Proven Technology
Uses helium as a tracer
gas
• Ideal due to its
inertness and small
atomic size
Defect detection down
to 2 µm
• Correlated to microbial
ingress studies
Restraining Supports
Pressure gauge

26. 26
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Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
1 Test Setup
The SU Assembly is
placed inside the
vacuum chamber
• Between the
restraining support
plates
• Connected to the
helium supply
P
Vacuum Chamber
SU Assembly
Restraining Supports
Air
Pressure gauge

27. 27
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P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
SU Assembly
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
2 Air Evacuation
The vacuum pump
valves are opened
The air inside the
chamber & assembly is
evacuated through the
exhaust lines
Restraining Supports
Air
Pressure gauge

28. 28
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P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
3 Helium Fill
The helium supply
valve is opened
The SU Assembly is
filled with helium
Restraining Supports
Helium
Pressure gauge

29. 29
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P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
4 Measurement
The helium detector
valve is opened
Restraining Supports
Helium
Pressure gauge

30. 30
Helium Integrity Test Technology | 31.March.2022
P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
4 Measurement
PASS
If the SU Assembly is
integral, all of the
helium will remain
inside the assembly
The helium detector
will not detect helium
Restraining Supports
Helium
Helium
Leak
Rate
Time
Threshold
Pressure gauge

31. 31
Helium Integrity Test Technology | 31.March.2022
P
Vacuum Chamber
Atmosphere
Helium Supply
Exhaust
Vacuum Pump
Atmosphere
Helium Detector
Exhaust
Vacuum Pump
High-sensitivity Helium Integrity Release Test
HIT Working
Principle
4 Measurement
FAIL
If there is a defect,
helium will leak from
the assembly
The helium detector
will detect helium
Restraining Supports
Helium
Helium
Leak
Rate
Time
Threshold
Pressure gauge

32. What to Expect With
mobius® Single-use
Assemblies Tested
With Helium Integrity
Test

33. Mobius® Single Use Systems and Certificate of Quality
What’s in the box?
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33
Certificate of Quality
Same as Gold level with different
Integrity Test Claim
Mobius® Single Use Systems
Double bagged
(No change to standard packaging)

34. Example Certificate of Quality
Mobius® Assembly, Gold with Helium Integrity Test
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100% Integrity Testing in Manufacturing
Each assembly was tested and passed an integrity
test method using helium as a tracer gas.

35. Mobius® Quality Certification Levels
Now even more options to meet your process needs
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Certification
Level
Shelf
Life
Sterility
USP <85> Endotoxins
USP <788> Sub-visible
particulates
Leak/Integrity
Testing
USP <88> Biological
Reactivity
In vivo (Class VI)
USP <661>
Physicochemical tests
Gold with
Helium
Integrity Test
2 years
Sterile
(Quarterly dose audit)
One assembly per lot tested
100% assembly
integrity tested
Post-gamma irradiation
Gold 2 years
Sterile
(Quarterly dose audit)
One assembly per lot tested
100% assembly
leak tested
Post-gamma irradiation
Silver 2 years
Sterile
(Quarterly dose audit)
Quarterly review of results
Lot sample
tested
Post-gamma irradiation
Bronze No claim No claim No testing performed
No testing
performed
Post-gamma irradiation

36. Product Manager, Final Fill Single Use Systems
charles.raye@milliporesigma.com
Senior Program Manager
monica.cardona@milliporesigma.com
Chuck Raye
Monica Cardona
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