CCI Overview and

USP 1207 Guidance

February 2021
Overview
• Defining CCI
• Regulatory Guidance and USP 1207
• Probabilistic vs Deterministic Methods
• Blue Dye Case Study
• Vacuum Decay Overview
• HVLD Overview
• Vacuum Decay vs HVLD

2
“A container and closure system
refers to the entirety of
packaging components that
together contain and protect
the product. “

FDA Guidance for Industry: Container and Closure System Integrity Testing in Lieu of Sterility
Testing as a Component of the Stability Protocol for Sterile Products, February 2008.

3
How to discuss integrity?
Sterile Product Packaging – Integrity Evaluation <1207>
Package integrity is defined as a package’s ability to prevent product loss, maintain
product sterility.

➢What is Important to the package?

➢What is important to the product?
➢What is important to your customer?
Package Integrity
Smallest leak to allow ingress determination
Lee Kirsch, et al, PDA J Pharm Sci & Technol, Vol. 51, No. 5, 1997

108 to 1010 P. diminuta E. Coli

From: MicrobeWiki From Wikipedia

• Glass micro-pipettes through wall of stoppered glass vial

• 0.1 to 10μm diameter (Sized via helium mass spec)
• Microbial challenge by immersion + liquid tracer element
• Challenge conditions
• Water bath immersion 60ºC 2hr, then 25ºC 1hr
• 24 hr immersion, ambient pressure
Most bacteria are 0.2 µm in diameter and 2-8 µm in length. The
three basic bacterial shapes are coccus (spherical), bacillus (rod-
Ingress Risk Dropped: Log -3.8 sccs (Leak < ~1μm) shaped), and spiral (twisted), however pleomorphic bacteria can
assume several shapes.
No Ingress: Log -5 to -5.8 sccs (Leak ~0.3 to 0.2μm)

5
Shift in CCI
• FDA Guidance for Industry (1999)- Container and Closure System
Integrity Testing in Lieu of Sterility Testing

• USP Chapter 1207 (2019)– Package Integrity Evaluation – Sterile

Products

• EMA Annex 1 (2020)– Manufacture of Sterile Medicinal Products

6
Modern Guidance: The Revised <1207>

• 3 pages to 4 subsections totaling ~50 Probabilistic Deterministic

pages • High Voltage Leak
• Bubble Emission
Detection (HVLD)
• Exhaustive list of available
• Laser-Based
• Microbial
technologies Headspace
Challenge
Analysis (HSA)
• Explicit preference for deterministic • Tracer Gas
methods (Helium) in Sniffer • Mass Extraction
Mode
• Does not support correlation to • Tracer Liquid (Dye) • Pressure Decay
microbial ingress • Tracer Gas
(Helium) in
• Broad lifecycle scope Vacuum Mode
• Vacuum Decay

7
Defining Deterministic
Deterministic: the leakage event is based on phenomena that follow a predictable chain of events, and
leakage is measured using physicochemical technologies that are readily controlled and monitored, yielding
objective quantitative data.

Probabilistic: its stochastic in nature in that it relies on a series of sequential and/or simultaneous events
each associated with uncertainties, yielding random outcomes described by probability distributions.

Deterministic methods Probabilistic methods

• Electrical Conductivity and Capacitance (HVLD) • Bubble Emission
• Laser-Based Gas Headspace Analysis • Microbial Challenge, Immersion Exposure
• Mass Extraction • Tracer Gas Detection, Sniffer Mode
• Pressure Decay • Tracer Liquid
• Tracer Gas Detection, Vacuum Mode
• Vacuum Decay
Multiple arrows in the quiver to
achieve container closure
integrity.

Not one test method alone can

achieve container quality
assurance.
Dye Ingress
DYE Ingress Test
• “Physical method” in old <1207>
• Samples are exposed to methylene blue • Likely the most common leak test
dye for a specific time under vacuum method
• Dye forced into the container due to the
pressure differential • Still probabilistic – hard to validate
• Look for presence of Dye in the container
• No standard approach

For example…

The following slides are adapted from a presentation given by Dana

Guazzo at the 2011 PDA Metro Chapter Conference based on work
performed by H. Wolf, et al, PDA J Pharm Sci & Technol., 63, 2009, p. 489
- 498

10
Dye Ingress Test Methods

11
Dye Ingress Comparison Test
• 1mL BD Staked Needle Syringes
• Consisted of negative controls and
positive controls with laser-drilled defects
of 5, 10, and 15 µm in nominal diameter
• Testing performed across different sites
and operators
• Operators had previously been qualified to
see dye at concentrations 0.2 - 0.5ppm
H. Wolf, et al, PDA J Pharm Sci & Technol., 63, 2009, p. 489 - 498

12
USP / Ph. Eur. Dye Test Results

H. Wolf, et al, PDA J Pharm Sci & Technol., 63, 2009, p. 489 - 498

13
ISO Dye Test Results

H. Wolf, et al, PDA J Pharm Sci & Technol., 63, 2009, p. 489 - 498

14
Modified ISO Dye Test Results

H. Wolf, et al, PDA J Pharm Sci & Technol., 63, 2009, p. 489 - 498

15
 Vacuum Decay Dye Ingress
Leak Test Results Visual Inspection Results2
Defect Type ID Code1
dP Pa P/F Inspector 4 Inspector 5 Inspector 6
B6 8 P N N N
B7 8 P N N N
Controls Tested
B8 8 P N N N
for Ingress
B9 8 P N N Y
B10 8 P N N N
111 64 F Y Y Y
112 54 F N N Y
5μm hole 113 88 F Y Y Y
114 56 F N N N
115 46 F N N Y
126 192 F Y Y Y
127 184 F Y Y Y
10μm hole 128 186 F Y Y Y
129 301 F Y Y Y
130 194 F Y Y Y
141 352 F Y Y Y
142 356 F Y Y Y
15μm hole 143 346 F Y Y Y
144 445 F Y Y Y
145 371 F Y Y Y

1 Holed syringes are identical to those used for Part 1, ASTM precision and bias studies.
2Y = dye seen, N = No dye seen

16
Dye Ingress Research
Vacuum Decay Container/Closure Integrity Testing Technology. Part 2. Comparison to Dye
Ingress Tests
Heinz Wolf, Tony Stauffer, Shu-Chen Y. Chen, Yoojin Lee, Ronald Forster, Miron
Ludzinski, Madhav Kamat, Brian Mulhall and Dana Morton Guazzo; PDA Journal of
Pharmaceutical Science and Technology September 2009, 63 (5) 489-498

Development of a Dye Ingress Method to Assess Container-Closure Integrity: Correlation to

Microbial Ingress
Lana S. Burrell, Mary W. Carver, George E. DeMuth and William J. Lambert; PDA Journal of
Pharmaceutical Science and Technology November 2000, 54 (6) 449-455

Comparing Physical Container Closure Integrity Test Methods and Artificial Leak
Methodologies
Sarah S. Peláez, Hanns-Christian Mahler, Christoph Herdlitschka, Toni Wertli, Matthias
Kahl, Atanas Koulov, Anja Matter, Satish K. Singh, Martina Widmer, Oliver
Germershaus and Roman Mathaes; PDA Journal of Pharmaceutical Science and
Technology May 2019, 73 (3) 220-234

Container Closure Integrity Testing—Practical Aspects and Approaches in the Pharmaceutical

Industry
Helen Brown, Hanns-Christian Mahler, James Mellman, Alejandra Nieto, Daniel
Wagner, Matthias Schaar, Roman Mathaes, Juergen Kossinna, Franz Schmitting, Sascha
Dreher, Holger Roehl, Markus Hemminger and Klaus Wuchner; PDA Journal of Pharmaceutical
Science and Technology March 2017, 71 (2) 147-162

17
Vacuum
Decay Leak
Detection

18
Vacuum Decay Test Method

• ASTM F2338-09 (2013) - Standard Test Method for

Nondestructive Detection of Leaks in Packages by
Vacuum Decay Method
• PTI’s VeriPac technology was utilized to develop this
test method
• Measures the level and change of vacuum over time
• Inspects the integrity of the entire package
• FDA Consensus Standard and USP 1207 Compliant
• Sensitive down to 0.01 cc/min 2 micron

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Vacuum Decay Test Cycle
Cycle 1 (step1 -2) Cycle 2 (step 3-5)
Large Leaks Small Leaks
Test Cycle
900.0

800.0

700.0
1 2 3 4 5
Absolute Pressure, mb

600.0

500.0
Test Cycle Pass
400.0
Large Leak
300.0 Small Leak

200.0

100.0

0.0
0 1 2 3 4 5 6 7 8 9 10
Test Cycle – Time in seconds

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Vaporization of Liquid Defects

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Liquid Defect Amplification

6R Vials - (Air leaks measured/verified at 500mb then filled

with WFI tested at 5mb)
dPa 450

400
500mbar
350

300
5mbar
250

200
0.86
150

100

0.22
50
0.05 Leak size [um]
0 Labels are MC flows in ccm
0 2 4 6 8 10 12
MC 500 mbar Real 500 mbar Real <10 mbar <10 mb out of scale

22
Leak Test Data

40
+3.42ccm
35 Master+3.42ccm

30
+1.94ccm 20um
25
dp, Pa

Master+1.94ccm dp Ref
20
15um
15

10

0
0 5 10 15 20 25 30
All data Avg Avg+3std

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Biologics and CCIT
• Smaller batches, higher value, sensitive products
• Wide array of product chemistries and structure
• Complex molecules
• “Proteinaceous product may interfere with defect
detection.”1
• Naturally occurring defects and laser defects are
susceptible to blockage when testing.
• Solutes may be deposited internal to the defect Albumin – 68,500 Daltons
passage during testing due to liquid vaporization

106

Sucrose – 342 Daltons

1 Li,
Lei. American Pharmaceutical Review. February 20, 2013. Container Closure Integrity
Testing Method Development and Validation for Prefilled Syringes.

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 High Voltage Leak
Detection (HVLDmc)
 Functional Principle of HVLDmc Test
Good Sample Leak

C1 – volume btw. C1
inspection electrode I1 I2
& the product V V
V R R

C2 C2
R – Liquid in the
C2 – volume btw. detection vial/syringe
electrode & the product
2 capacitors 1 capacitor

V – High Voltage Source V = IR V = I ↑ R↓

R – Electric Resistance of the product
C1 – Capacitor 1: Glass between the inspection electrode and product
C2 – Capacitor 2: Glass between the detection electrode and product
I1 – current produced when product container is sealed
I2 – current produced when product container is defective
High Voltage Leak Detection (HVLDmc)

e-

Inspection
Electrode
C1  Glass Wall
C1
R  Liquid Contents

C2  Glass Wall

Detection
Electrode

27
High Voltage Leak Detection (HVLDmc)
Conventional vs Microcurrentmc

This new technology applies less than 50% of the voltage used with conventional high
voltage technologies and the product its exposed to less than a 5% of the voltage exposure
experienced when testing with comparable HVLD solutions.

Conventional HVLD
Pure AC voltage:
VAC i
C1

VDC DC offset
-16kV
C2

AC Voltage
5kVpp

MicroCurrent HVLD
AC + DC voltage:

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MicroCurrent HVLDmc
• The MicroCurrent applied to the product during the test greatly reduces the
voltage exposed to the product and environment.
MicroCurrent HVLDmc
• The MicroCurrent applied to the product during the test greatly
reduces the Ozone Production.
MicroCurrent HVLDmc
• The nature of this solution allows the testing of packages with extremely low
conductivity liquids such as sterile water (WFI).

Source: PTI / Packaging Technologies and Inspection

Vacuum vs. HVLDmc

• 1mL and 2.25 mL Syringe

• Positive Controls: 5, 10 und 20 μm laser drilled holes, barrel
and shoulder
• 3 positive controls with water for each size
• 5 positive controls with Albumin for each size
• Albumin concentration of 17.5%
• Two test methods: Vacuum Decay and HVLDmc
Vacuum Decay – 1mL Syringe
Vacuum Decay – 2.25ml Syringe Case Study 2

Only 3 H2O positive controls

clearly identified
HVLD 1mL Syringe

All positive controls identified

HVLD 2.25mL

One unidentified positive control

Summary
• Know your package and what attributes can effect your
product over the product life time.
• Current Guidance for CCI points towards Deterministic
Test Methods
• No Golden All Encompassing Test Method to cover all
package/product combinations.
• Understanding the risks over the product life cycle and
mitigating those risks through strategy/ analysis is key.
Supplemental Slides

39
 Realistic Defects

COPYRIGHT © PDA 2018

 Naturally Occurring Defects

Sample leak rate Visual Size

mbar·l/s 10-5 mbar·l/sec sccm um

1 0.00095 95 0.05700 Large crack 2.67

Sample 2 Sample 6
2 0.0000021 0.21 0.00013 Medium crack 0.13

3 0.000014 1.4 0.00084 Medium crack 0.32

4 0 0 0.00000 Small Scratch 0.00

6 0.00067 67 0.04020 Large crack 2.24
7 0.015 1500 0.90000 Large crack 10.61
Sample 3 Sample 16
9 0.00000029 0.029 0.00002 Small crack 0.05
10 0.00029 29.0 0.01740 Large crack 1.47
11 0.074 7400 4.44000 Large crack 23.56
12 0.055 5500 3.30000 Large crack 20.31
16 0.0014 140 0.08400 Small crack 3.24
18 0 0 0.00000 Small Scratch 0.00
19 0 0 0.00000 Small Scratch 0.00
20 0.016 1600 0.96000 Large crack 10.95

COPYRIGHT © PDA 2018

HVLD Natural Defects
He Certified Leak Rate
❖ Controlled crack produced. 95.0 10-5 mbar·l/s
Sample 1
❖ Certified flow measurement with Helium 0.05700 sccm
mass spec. 2.67 ~µm
❖ Peak signal response is recorded.
❖ Voltage signal response with each rotation.

10.0V

COPYRIGHT © PDA 2018

HVLD Natural Defects

He Certified Leak Rate He Certified Leak Rate

Sample 6 67.0 10-5 mbar·l/s Sample 3 29.0 10-5 mbar·l/s
0.04020 sccm 0.01740 sccm
2.24 ~µm 1.47 ~µm

10.0V 10.0V

COPYRIGHT © PDA 2018

HVLD Natural Defects
He Leak Rates
He Certified Leak Rate He Certified Leak Rate
Sample 2 0.21 10-5 mbar·l/s Sample 3 1.40 10-5 mbar·l/s
0.00013 sccm 0.00084 sccm
0.13 ~µm 0.32 ~µm

10.0V 7.6V

COPYRIGHT © PDA 2018