Toxikon

Extractables & Leachables
Training
Michael Ruberto John Iannone
President & Program Manager
Material Needs Consulting, LLC Toxikon Corporation
1
The Challenge
• Ensuring the safety and compatibility of

materials used in medical devices and
pharmaceutical packaging / processing
equipment
– Metal, Glass, Polymers
Who must meet the challenge?
Pharmaceutical Medical Device
Suppliers
CCS Vendors
Equipment Vendors
Resin Manufacturers
Container Closure Systems
4
Medical Devices
5
Processing Equipment
6
Materials Used in Medical Applications
• Commonalities
– Materials
• Plastic, Rubber, Glass, Metal, Ceramic
– Polymers are More Prone to Migration of
“Leachables” than Traditional Materials Such as
Metal or Glass
• Differences
– Risk – Depends on End Use
– Regulations
– FDA Jurisdiction
– Other Regulatory Bodies
7
Regulations
Where do we start?
8
FDA Centers
FDA Centers with
Product Responsibilities
Center for Devices
and Radiological Health (CDRH)
Medical Devices Center for Veterinary
Medicine (CVM)
Center for Biologics

Evaluation and Research (CBER)
Medical Devices and Packaging
Center for Food Safety

and Applied Nutrition (CFSAN)
Center for Drug
Evaluation and Research (CDER)
Medical Devices and Packaging
FDA oversight for medical devices authorized by 1976 Medical Device

Amendments of 1976 (§513, FD&C Act, as amended)
9
What is a Medical Device?
Per Chapter 2, §201(h) of Food, Drug, and Cosmetic Act, as amended:
• “A medical device is an instrument, apparatus, implement, machine,

contrivance, implant, or in vitro reagent”
• Intended to: - Diagnose diseases/conditions;

- Cure, mitigate, treat, or prevent disease;
- Affect the structure/function of the human body; and
• Does not achieve primary purpose through chemical action in or on the

body.
• Is not dependent upon being metabolized for its primary purpose.
Primary characteristics that distinguish medical devices from drugs.
10
What about combination products?
• CBER Jurisdiction:
•Fibrin Sealant (and delivery system)
•Intranasal Flu Vaccine
• CDER Jurisdiction:
•Metered Dose Inhalers
•Transdermal Patches
• CDRH Jurisdiction:
•Fibroblast-derived Wound Dressing
•Drug Eluting Coronary Stent
11
Questions Regarding Materials
• Patient Safety
• Adulterating the Product
– Efficacy
• Protection
• Performance
• Concern:
Will chemicals in the materials leach or
migrate into the drug product or patient?
12
Extractables and Leachables
Chemicals that can migrate from CCS, devices and

processing equipment into the drug product
• Extractables – Potentials
– Determined in Laboratory Experiments using Neat Solvents
under Conditions that Predict the “Worst Case” Conditions
of Exposure.
• Contact Time, Temperature, Solvating Power of Drug. etc
• Leachables – Actuals
– In the Presence of the Drug or Drug Components During
Normal Use
• Processing, Shelf-Life, Stability Studies, etc.
All Materials Can Produce Leachables!
13
How to Deal with E&L
• Medical Devices
– Premarket Notification - 510(k)
– Premarket Approval - PMA
• Processing Equipment
– Usually Considered to be Drug Product Impurities
– BPSA (Bioprocess Systems Alliance)
• Pharmaceutical Packaging – Container Closure
Systems
– E&L are Linked to Packaging
– Best Practices are Beginning to be Developed
Can best practices developed from one
application be applied to others?
14
Whose responsibility is it?
The Submitter / Marketer of the Drug or Device

• Role of the Vendor – Facilitator
– Shorten Time to Market
• Documentation / Data
• Testing
– Ensure Sustainability of Product
• Change Control
• Reliable Supply Chain
15
The Issue
Risk
vs.
Reward
What is the proper balance?
16
Let’s Start with the Reward
• New Drug or Device Approvals!!!
17
Reward
…and Sustainable Regulatory Compliance

• Secure Supply Chain for Raw Materials
• Change Control
• No Unexpected Leachables
– Product Performance
– Product Safety
– Product Aesthetics
– Product Recalls
– OOS Issues
They Can’t Celebrate Prematurely
18
Risk
• Will they get the new drug or device approvals?

– Think Globally
• Will they do the appropriate amount of testing?
– Too Much vs. Too Little
– Resources
• What will be their time to market?
– Speed vs. Sustainability
• Did they make the correct decisions?
– Only Time Will Tell – 5 – 10 Years
19
Question???
• A Pharmaceutical Company Submits an NDA

for an Inhalation Product Delivered with an MDI
– FDA Approval with No Questions Regarding E&L
– Cost of E&L Studies - $2.0 Million
– Time to Perform Studies – 1.5 Years
• Was it a good Decision?

– Pros and Cons
20
There is No Correct Answer
• Must Balance Risk vs. Reward

• Regulations – Vary
• E&L “Best Practices” – Not Prescriptive
• My Personal Approach
– Conservative on the issues, but creative in
addressing them
• What are your customer’s approach?
– Any trends from small or large companies?
21
Materials Overview
22
Why Polymers?
•Light-weight
•Flexible
•Durable
•Clear
•Color Enhanced
•Unique Shapes
•Disposable
23
Polymers are melt processed….
•Temperature
• Shear
• Exposure / Duration
 Degradation
Processing can significantly affect the polymer – what comes out may
be different than what went in….
Photos Courtesy of Ciba
Polymer Processing
Addition of Additives
Complex Formulation - Leachables
25
Questions
• What is the composition of my component?

• What is the impact of this material change?
• Should I accept the vendor supplied extractables
data?
• How do I design my controlled extractables testing?
• What are these unknown peaks?
• How much “control” do I have over my supply chain?
• What type of QC system should I implement to accept
new lots of processing equipment?
26
Answers
An understanding of the chemical and

physical properties of the materials
can provide answers to these
questions!!!
27
Important Concepts to Consider
• Solubility of Chemicals in the Polymer

– “Like Dissolves Like”
– Reduces Exudation and Blooming
• Diffusion / Migration out of the Polymer
– Thermodynamics and Kinetics
• Impact of External Conditions
– Shipping and Storage
– Sterilization
– Contact with Drug
• Compatibility
How to Simulate in the Laboratory
Time, Solvent, Temp.
28
Polymers
A polymer is a large macromolecule

composed of many smaller repeating
Units containing C, H, O, N, Si, Cl,
Fl, Br, P, or S
• Linear or Branched Framework

• MW Can A Few Thousand – Several Million g/mol
29
Main Groups of Polymers
Elastomer Thermo set

bridges bridges
polymer chain
polymer chain
Thermoplast
•high temperature resistance
•high temperature resistance semi crystalline amorphous •high chemical resistance
•high abrasion resistance crystalline part •very high tensile strength
•high flexibility
•not meltable
amorphous part
•opaque •transparent
•Melting point and glass point •glass transition point
•“defined melting point” •brittle
•high tensile strength •undefined flow temperature
•high E-modulus range
30
Factors Affecting the Mechanical Properties
of Polymers
• Melting Point, Tm
• Glass Transition Temperature, Tg
• Degree of Crystallinity
• Amount of Cross-linking
31
Polymer Blends
Physical mixture of two or more different types of polymers
To blend different polymers is an easy way to improve certain

properties. The most popular blends are PC/ABS , PP/EPM
(high impact resistance), PA/PE (high impact resistance also
under dry conditions),
32
Copolymers
Examples: PP-Copo, ABS, POM,…
Monomer (Particle) A Monomer (Particle) B
Random-
Block-
Graft-
33
Reasons for Crystallization
Structural Regularity with Little Branching

Key - Lock principle: e.g. different types of PP
Molten phase
Compact, Ordered
Morphology
Solid phase
34
Reasons for Crystallization
Electronic Interactions
Hydrogen bonds: PA, POM
Each hydrogen bond has a contribution to the stability of the polymer

up to 4 kJ/mol.
35
It is not just…Polyethylene
LD-PE Density: 0.915 - 0.940m [g/cm3]

Mp: 105°C
Crystallinity up to 50%
Polymerization with peroxide
good aging resistance
Good resistance against chemicals
Density: 0.89 - 0.94 [g/cm3]
LLD-PE Polymerization with Ti/Al, Cr- or Metallocene-
catalyst
Stability depending on the cat. system
Density: 0.940 - 0.965 [g/cm3]

HD-PE Mp: 130°C - 146°C
Polymerization with Ti/Al, Cr-catalyst
Stability depending on the cat. system
High resistance against
UV-light
36
What Type of Polypropylene?
Density of 0.907 [g/cm3]; Crystallinity up to 70%, Mp: 160°C - 208°C;

Polymerization with Ti/Al, Cr or Metallocene-catalyst; Less stable against UV-
light then PE; good chemical resistance.
homo-PP:
isotactic n * highest crystallinity
syndiotactic n *
atactic n most transparent

The tacticity is determined because of the catalyst which is used for the polymerization
copo-PP:
raco
*
block * n m
37
Useful Polymer Information
• Polymer Type
– Homopolymer, Copolymer, Blend, etc.
• Tg and/or Tm
• Molecular Weight Distribution
– Monomers and Oligomers
• % Crystallinity
• Density
• Raw Materials
– Any Chemicals of Concern
• Impacts the Migration Rate of Leachables!
38
What is an Additive????
• Additives are Ingredients Incorporated into the

Polymer to Stabilize or Enhance its Performance
– Stabilizers – Maintain the Polymer’s Original Properties
• Strength
• Flexibility
• Toughness
– Modifiers – Change or Improve Polymers Performance

• Performance Additives (Slips, Anti-Stats)
• Pigments
• Fillers
39
Examples of Chemical Additives
CH3
CH
CH3 CH3
O
H O P O
HOOC H
CH3
40
What’s in a name?
A b u n d a n c e
T I C : 0 8 1 9 0 3 0 5 . D
3 2 0 0 0 0 0
3 0 0 0 0 0 0
2 8 0 0 0 0 0
Abietic acid
2 6 0 0 0 0 0
2 4 0 0 0 0 0
2 2 0 0 0 0 0
2 0 0 0 0 0 0
1 8 0 0 0 0 0
1 6 0 0 0 0 0
1 4 0 0 0 0 0
1 2 0 0 0 0 0
1 0 0 0 0 0 0
8 0 0 0 0 0
6 0 0 0 0 0
4 0 0 0 0 0
2 0 0 0 0 0
0
5 . 0 0 1 0 . 0 0 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 3 0 . 0 0
T i m e - - >
41
Stabilizer Classes

Melt Processing
Process Stabilization /
• Phosphites
Base Stabilization
• Hindered Phenols

Long-term Thermal
• Hindered Phenols
Thermal Stabilizers
• Hindered Amines

Light Stabilizers
• Radical Traps Light Stabilization
• UV Absorbers Package
• Excited State Quenchers
Used in Both Plastic and Rubber

42
Principle Structure of Main Antioxidants
Phenols: Phosphites :
Active group R'
O
OH R O P
High hindrance
O
R''
Low hindrance (RO)3P
R
Other Properties Phosphites consume

hydroperoxides before
Phenols act as hydrogen donor hydroperoxides decompose
and radical scavengers into free radicals
43
Degradation and Stabilization
R-H Melt Processing

(Polymer) Energy (UV Light, Heat)
Catalyst Residues
React with primary Oxygen
antioxidants to
yield inactive
products
(ROH and H20) Cycle II Cycle I
R•
RO • + • OH ROO •
Reacts
R• with
+ primary
anti-
ROOH oxidants
Path of R• Alkyl radical
Degradation RO • Alkoxy radical
Reacts with secondary ROO • Peroxy radical
Path of antioxidants to yield ROOH Hydroperoxide
Stabilization inactive products (ROH)
44
Phosphites – Transformation Chemistry
O O
O O
P [O] P
O
O
[1]
45
Phosphites – Degradation Chemistry
OH
O
O P
O OH
P [ H2O ] O
O
+
[ H2O ]
OH
OH
OH OH
HO
[ H2O ] O P
P + 2
+ 3
OH
OH
46
Reaction of Hindered Phenol
with Singlet Oxygen
O OH O O
(CH3)3C C(CH3)3 (CH3)3C C(CH3)3
H3C OOH
CH3 H3C OOH H 3C OOH
OH OH O OH
(CH3)3C C(CH3)3 (CH3)3C C(CH3)3
OH OH 1O
2
(CH3)3C C(CH3)3
CH2OCH3 CH3 H3C OOH CH2OCH3
CH3 CH3 OH OH
O OH
(CH3)3C C(CH3)3
(CH3)3C C(CH3)3
CH2OCH3 CH2OCH3 H3C OH CH3
O O
(CH3)3C C(CH3)3
H3C OH H3C OH
47
Representative Commercial Phenolic AO’s
O
O
HO CH2 CH2 C O CH2 C
HO CH2 CH2 C O C18H37 OH
4
OH
HO O N O
O
N N
HO CH2 P O Ca2+
OC2 H5 O
HO
2
OH OH
OH H CH3 C8H17
OH OH
S OH
O
C8H17
HO (CH2)2 C N S
H O
2 HO CH2 CH2 C O (CH2)2-]2-S-
Creating Effects with Surface Additives
A controlled incompatibility in the polymer

• Antimicrobials
• Slip Agents and Lubricants
 Antistatic Agents
More Susceptible to Leaching
Wiping
or Migration
Washing
hydrophilic head
hydrophobic tail
49
Slip Agents
• Friction between surfaces of two films

• Friction between film surface and substrates
• Migration of slip agents towards the surface
• Controlled incompatibility with the polymer
50
Case Study- Material Change
Slip or Mold Release Agent
Vendor Replaces Erucamide with Oleamide
NH2
Erucamide
- migrating slower O
- lower vapor pressure
- less volatile
- used for high temperature processing conditions
NH2
Oleamide O
- migrating faster
- slip effect after very short time
- lower vapor pressure
- more volatile than erucamides
Possible change in leachables profile!

51
Color
Color can be very important in the marketing of a product.

Ensuring color consistency and quality in each
component of a container closure system can sometimes
require the use of multiple pigments when differing
polymer resins are utilized.
52
Other Stabilization Needs for Rubber
• Acid Scavengers
– Neutralize Halogen Anions in Rubbers
• Halogenated Isobutylene Isoprene
• Magnesium Oxide
• Hycite® 713
• Metal Deactivators
– Rubber Poisons
– Cu, Fe, Co, Ni, and other Transition Metals
– Catalyze Hydroperoxide Decomposition of Rubber
O
HO (CH 2)2 C NH Irganox® MD 1024
2
53
Extractables Profile for Plastic and Rubber
• Monomers or Oligomers from Incomplete Polymerization

• Additives
• Transformation and Degradation Products
• Lubricants / Surface Modifiers
A b u n d a n c e
•
T I C : 0 2 2 1 0 3 0 2 . D
Fillers 1
1
. 1
1
. 2
. 1
e
e
+
+
0
0
7
•
1 . 0 5 e + 0 7
Rubber Curing Agents 9
9
5
0
1
0
e
0
+
0
0
0
7
•
8 5 0 0 0 0 0
Vulcanizates 8
7
0
0
0
0
0
0
0
0
0
0
0
6 5 0 0 0 0 0
• Special Case Compounds 6
5
0
0
0
0
0
0
0
0
0
0
0
4 5 0 0 0 0 0
– PNAs 4
3
0
0
0
0
0
0
0
0
0
0
0
2 5 0 0 0 0 0
– Nitrosamines 2
1
0
0
0
0
0
0
0
0
0
0
0
5 0 0 0 0 0
– Mercaptobenzothiazoles T im e - - >
0
5 . 0 0 1 0 . 0 0 1 5 . 0 0 2 0 . 0 0 2 5 . 0 0 3 0 . 0 0 3 5 . 0 0
Courtesy of PQRI
54
Extractables and Leachables
Challenge
• Stabilization is a dynamic process
• Many additives will transform and/or degrade
during use.
– Transformation and degradation products must
be considered along with the intact additives as
potential leachables and extractables.
• Unexpected Additives May also be Present in
the Polymer
55
Polymer Supply Chain for CCS
Change Control Change Control
Monomer Polymer Manufacturing Masterbatchers End Products Pharma

Manufacturing Converters N-1 Suppliers
Synthesis Extrusion
Medical Grade
Resins Available
Process Polymerization Antioxidants Light

Chemicals Regulators Processing Stabilizers Stabilizers
Storage (Light stabilizers) (AO/Proc.
Stabilizers stabilizers)
56
How do we manage this situation?
• Additional Issues
– New Source for Raw Materials
– Polymers or Additives Discontinued
• Planned or Surprise
– Unexpected Component in the Formulation
Upstream Change Control Issues
57
Unexpected Changes
Greater Sales Volumes to Other Markets
Cleaning
Personal Care
Polymer Manufacturing Masterbatchers

Monomer
Manufacturing Converters
Synthesis Extrusion
Food
Can Drive Upstream Changes

Automotive
Outdoor
58
Masterbatchers and Converters
Polymer Concentrate
-Stabilizers
-Pigments
-Antistats
Polymer Often Contains a Base Stabilization Package
59
Metal - Deep Drawing Process
Deep-
Drawing
tool
Metal Rolls
Images provided by Presspart
60
Metal - Deep Drawing Process
finished canisters
Images provided by Presspart
Degreasing Process
61
Extractables from Glass
Generally has better barrier properties than plastic,

however…
• Extractables
– Alkali Oxides
– Trace Metals
– Silicone – Assists in plunger glide
• Interactions with Drug Products - Biologics
– Higher pH DP can Increase Leachables
• Particulates
– pH Shifts
– Protein and Peptide Aggregation
– Tungsten Issues
62
Overview of E&L
“Best Practices”
Extractables and Leachables Testing
What is PQRI?
• Product Quality Research
Institute
• Not-for-profit, non-stock, tax-
exempt entity incorporated in
Virginia
• Serves as a forum for
academia, industry and FDA
to work cooperatively
• Working Groups on
Leachables and Extractables
currently in operation
• OINDP – Submitted Best
Practices
• PODP – In Progress
64
Establish an E&L Program
Partner With Vendors Manage Supply Chain
Extractables Leachables
Quality
Control
65
Extractables and Leachables Testing
• Controlled Extraction Study 14

Extraction Efficiency
– Multiple Solvents of Different Polarity 12
10
Amount (Micrograms)
– Commonly Used Extraction Techniques
8
• Soxhlet or Reflux 2
0
0 0.5 1 2 4 16 24
Time (Hours)
• Leachables Study
– Qualitative and quantitative analysis of the migrants from
a container closure system or device into a drug product
over the course of its shelf-life
– Correlation to Extractables Profile
• Routine Extraction Testing
– Optimized Controlled Extraction Conditions
– Quality Control
66
What about the evaluation of leachables
from other dosage forms?
• Issue: The safety evaluation thresholds,

analytical testing, and best practices
recommendations were developed for
OINDP.
• They are currently being applied across
the board to other dosage forms as well
as non-packaging applications
– Implantables, Processing Equipment, etc.
67
Differences in Materials - MDI
• Metal
– Stainless Steele
– Aluminum
• Plastic
– PE
– PP
• Elastomers
– Various Types of Rubber
68
Differences in Materials - Parenterals
• Plastic
– PP, COC, PVC, PC
• Overwraps
– LDPE
• Rubber
• Surface Treatments
– Lubricants
• Printing / Ink
69
Comparison of Formulations
Acidic
PODP
MDI
Polar Basic Non-polar
70
Comparison Between OINDP and DODP
Daily Dose
MDI or DPI LVP – Large Volume

Small Volume Small Number of Doses
Large Number of Doses
71
Consider the Safety Assessment Triad
Material Characterization
(Controlled Extraction Study);
Screening and Selection
Extractables as tentative leachables
Simulation Study
(Simulated Extraction Study)
Worst-Case Safety Assessment
Extractables as probable leachables
Potentially Useful
For LVPs Migration Study
(Target Leachables Study)
Actual Case Safety Assessment
Confirmed leachables
D. Jenke PQRI PODP Workshop Feb. 22-23, 2011

E&L For Processing Equipment
Regulatory Comments •
“…with respect to extractable and leachable data …it is ultimately your
responsibility to assess this data and it’s applicability to your products and process.
CBER recommends a risk‐based approach be taken in evaluating extractables
and leachables …
where you take multiple aspects into account (e.g., indication, safety issues, product
characteristics, dosage, formulation, stability profile, etc.).
If there is no relevant risk associated with the…

(material in question)…, vendor data could be cross referenced
and a detailed justification for the applicability of this data and a justification for no
additional testing should be submitted. “
–Destry M. Sillivan, FDA CBER

IBC Single Use Conference, 2010
CBER/DMPQ communication to regulated industry
E&L For Processing Equipment
• Bioprocess Systems Alliance (BPSA)

– Extractables 101 (2008)
– Extractables 102 (2010)
• Evaluate Vendor Supplied Information
– COAs
– Extractables Data
74
BPSA Risk Assessment
• Make a List of the Components in the Processing Scheme

that Contact the Drug
• Consider the Following:
– Material Compatibility
– Drug / Solution Solvating Strength
– Proximity to Final Product
– Surface Area / Volume Ratio
– Contact Time and Temperature
– Pretreatment of Components
• Sterilization, Flushing, etc.
75
BPSA Recommended Extraction Conditions
• Use Exaggerated Time, Temp, Surface Area / Volume

Ratio, and Worst Case Pre-Treatment Steps
• Solvents – At Least 2
– Water, Alcohol, Hydrocarbon, Oil
• Temperature
– Elevated with Respect to Actual Use Conditions
– Avoid Extremes
• Time – Slightly Longer than Actual Contact Time
• Dynamics – Mimic Actual Use
– Static vs. Flowing System
76
BPSA Continued
Migrants vs. Leachables

• Migrants – “Chemical Species Derived from Process
Equipment in Actual Use”
– Ensure “Worst Case” Conditions
• Leachables – Packaging Term
• Analysis of Extractables and Migrants

– GC, HPLC, ICP, FTIR
77
Case Study – Applying the Concepts
Pt Cured Silicone Tube
78
Material Assessment (1)
• Vendor Supplied Information

– USP Class VI
– EP 3.1.9
– FDA CFR 177.2600
– Compositional Information & Migration Data
Example CFR 177.2600 – Rubber Articles Intended for
Repeated Use
– For Aq Foods must produce less than 20 mg/in2 of extractables
after water reflux for 7 hrs
– For Fatty Foods must produce less than 175 mg/in2 of extractables
after hexane reflux for 7 hrs
79
• Pt Cured Silicone
– Low Level of Pt Catalyst (10 ppm)
– No By-products
– Long Chains with Minimal Crosslinking and Low Tg
• Elastic at Ambient T, so no need for Plasticizers
– No Antioxidants, Heat, or Light Stabilizers Required
– Amorphous Polymer with Inadequate Mechanical
Properties
• Inorganic fillers Added
80
• Possible Leachables
– Low MW Monomers and Oligomers
• Polydimethylsiloxane (PDMS)
– Straight Chain or Cyclic Siloxanes Si O
O
– Inorganic Fillers and/or Pigments

Si Si
• Impurities in these Fillers

– Sorption can be a Problem if Filling Lines are
Left Idle for a period of Time
Si O O Si
O Si Si O
Si O
O Si
O Si Si O
Si O O Si
O Si
81
Pt Cured Silicone Tubing Case Study

• Upstream Transfer Line
• Ambient Temperature
• Contact Time < 1 hr
Solvent Possible Migrants Risk

Aqueous Mostly Inorganics Low
Aqueous Buffer w/ 20% Inorganics, Siloxanes, Moderate
Tween 80 Monomers
Oil Based or High Organic Monomers, Siloxanes High
82
Materials Assessment (2)
• Evaluate the Risk

– Low Risk
• Assessment Report
– Moderate Risk
• Assessment Report and/or Extractables Testing
– High Risk
• Assessment Report and Extractables Testing
• Possible Leachables Testing
83
Extractable/Leachable
Testing Considerations
Testing Needs
Testing Needs starts with understanding Needs for Testing
E&L Testing: Determining the Interactions between

Pharmaceutical Containers and Drug Products
Can have an impact on:

o Drug Product Efficacy
o Drug Product Safety
o Drug Product Compatibility
For Primary Packaging

For Materials in (Bio)Pharma Production
Reminder
• Extractables:
Extractables are compounds that migrate from the contact surface
under more aggressive conditions such as elevated temperature,
extended contact time, or aggressive solvent system. Any component
that is added to the device or the materials used to make the device.
What CAN come out.
• Leachables:
Leachables are compounds that migrate from the contact surface
under normal conditions of exposure. Leachables are usually a
subset of extractables.
What DOES come out.
86
Focus
The terms extractable and leachable provide clarity in terms of:
1. The potential versus the actual impact of the product on its user.
• Extractable = potential impact: what “could” come out
• Leachable = actual impact: what “will” come out
2. The object on which the testing is performed.

• Extractable = test the material
• Leachable = test the finished product
D. Jenke (presentation at SmithersRapra, Providence, May 2013)
87
Relationship
Extractables/Leachables
• LEACHABLES are typically a SUBSET of EXTRACTABLES
• NOT ALL LEACHABLES are EXTRACTABLES
88
Regulations
REGULATORY ASPECTS – PARENTERALS – NON-LIMITATIVE LIST
<1999: 21CFR 211.94(a) “DRUG PRODUCT CONTAINERS AND CLOSURES”

...not reactive, additive, absorptive to alter
safety, identity, strength, quality or purity of drug...
1999: “CONTAINER/CLOSURE SYSTEMS FOR PACKAGING

HUMAN DRUGS AND BIOLOGICS” (FDA-Guidance for Industry)
2003: EU COMMISSION DIRECTIVE 2003/63/EC, (§ 3.2.2.2 g)

• CCS-information is part of the Market Authorization dossier.
2005: “GUIDELINE ON PLASTIC IMMEDIATE PACKAGING MATERIALS”

(EMEA Guideline)
• Contains “Decision Tree” for different dosage forms
2006: ICH Q8 “PHARMACEUTICAL DEVELOPMENT”, §2.4 CCS
2006: “PQRI Safety Thresholds and Best Practices for E/L in OINDP’s”
• First technical recommendations for E/L-studies!
• Threshold concept!!
Regulations
REGULATORY ASPECTS – PARENTERALS – NON-LIMITATIVE LIST
<1999: 21CFR 211.94(a) “DRUG PRODUCT CONTAINERS AND CLOSURES”
...not reactive, additive, absorptive to alter safety, identity, strength, quality or purity
1999: “CONTAINER/CLOSURE SYSTEMS FOR PACKAGING

HUMAN DRUGS AND BIOLOGICS” (FDA-Guidance for Industry)
• Classification, based on likelihood of interaction and route of administration
2003: EU COMMISSION DIRECTIVE 2003/63/EC, § 3.2.2.2 g)
• CCS-information is part of the Market Authorization dossier.
2005: “GUIDELINE ON PLASTIC IMMEDIATE PACKAGING MATERIALS”

(EMEA Guideline)
• “Decision Tree” what information to provide for different dosage forms
2006: ICH Q8 “PHARMACEUTICAL DEVELOPMENT”, §2.4 CCS
2006: PQRI Safety Thresholds and Best Practices for E/L in OINDPs
• First technical recommendations for E/L-studies!
• Threshold concept!!
Regulations
Pre-Filled
Syringes
The FDA Guidance Document

“Container Closure Systems for Packaging Human Drugs and Biologics” of 1999
91
Regulations
The FDA Guidance Document
“Container Closure Systems for Packaging
Human Drugs and Biologics” of 1999
may NOT reflect the current (2013) FDA requirements
for E/L Testing and Documentation:
o Requirements for Freeze Dried Products = Liquid Parenterals
o NOT ONLY EXTRACTABLES evaluation => Consider LEACHABLE STUDIES!
o Consider All Container Parts of a Pre-Filled Syringe

Not only parts that are in Contact with DP
Also Needle Shields/Tip Caps
Glue Residues, Tungsten Residues, Silicone oil
Secondary Packaging, if necessary
92
Regulations
The EM(E)A Guideline on “Plastic Immediate Packaging Materials” of 2005
Pre-Filled Syringes
93
Regulations
The EM(E)A Guideline on

“Plastic Immediate Packaging Materials” of 2005
may NOT reflect the current (2013) European Regulatory Requirements
for E/L Testing and Documentation:
• Not for Elastomers (?) = > In reality: ALSO fo rubbers
• If a Material is described in the E.P. And if it complies with the specifications therein, no
Extractable testing may be needed.  NOT ACTUAL POSITION OF EU REGULATORS
• If Extractable Testing shows only compounds with low risk (at low concentrations) no leachable
study is necessary.  NOT THE ACTUAL POSITION OF EUROPEAN REGULATORS
94
Extractables / Leachables
The Challenges in E/L testing
Challenges in E/L-Testing
Diversity of non-API Related Compounds
in E/L research is Tremendous!!
Broad spectrum of:

o Types of Containers
o Types of Materials used in the Manufacture of Containers
o Number of Suppliers per Material
o Number of Grades (per supplier) for each type of Material
o Type of Sterilization (impact on material impurity profile)
CONFIDENTIAL 96
(Non Limitative) List of types of Pharmaceutical Containers
INHALATION PARENTERAL SECONDARY PACKAGING
o Metered Dose Inhaler Components o Bottles o Labels
• Gaskets o Vials o Adhesive/Glue (e.g. on labels)
• Stem o (Pre-Filled) Syringes
• Body
o Ink
o Cartridges o Overwrap foils
• Metering Chamber
• Protection Ring o (Rubber) Stoppers o Blisters
• Actuator o Rubber Plungers o Cardboard packaging
• Cannister o Needle Shields
o Dry Powder Inhaler Components o Tip Caps MEDICAL DEVICES
o Nasal Spray Systems o I.V. Bags o Stents
o Nasal Dropper Systems o Administration Sets o Cathethers
o I.V. Bags
OPHTHALMIC SINGLE USE SYSTEMS o Admiunistration Sets
o Eye Dropper Systems o (Multilayer) Bags o Implantation Devices
o Tubes o Tubings
o Connectors
DERMAL/TOPICAL o Ports
o Spray Systems o Filters (+ Housing)
o Tube systems o Chromatographic Columns
o Lyo trays
97
Pharmaceutical Containers can be made of different Materials
o Low Density Polyethylene o Polyamide (Nylon-6, Nylon-66)
o High Density Polyethylene o Cyclic Olefin Copolymers (COC)
o Polypropylene o Cyclic Olefin Polymers (COP)
o Rubbers o Polyethylene Terephthalate (PET, PETG)
o Butyl Rubbers o Polybutylene Terephthalate (PBT)
o Chlorobutyl Rubbers o Polyacetal (POM)
o Bromobutyl Rubbers o Polymethylmethacrylate (PMMA)
o EPDM Rubbers o Acrylonitrile Butadiene Styrene (ABS)
o Isoprene Rubbers o Silicone
o Nitrile Rubbers o C-Flex
o Latex Rubbers o Polycarbonate
o Other Rubbers o Teflon
o Multi-layer Films and Foils o PEEK
o Polyurethane (PU) o Glass
o Ethylvinyl Acetate (EVA) o Metals
o Ethylvinyl Alcohol (EVOH)
98
Each Material has different Suppliers
EXAMPLES
Polyethylene - produced by: Pharmaceutical Rubbers - main Global Suppliers:

o Borealis o Datwyler
o LyondellBasell o West Pharmaceutical
o SABIC o Stelmi
o Dupont
o Enichem
o INEOS
o TOTAL
Each Supplier has different Different Grades!
99
Each Supplier has different Different Grades
EXAMPLES
PolyEthylene - produced by:
o Borealis: over 30 different Medical Grades
o LyondellBasell: over 30 different Medical Grades
o SABIC: over 30 different Medical Grades
o Dupont: different grades
o Enichem: different grades
o INEOS: different grades
o TOTAL: different grades
Pharmaceutical Rubbers - main Global Suppliers:

o Datwyler: over 100 different commercial rubber formulations
o West Pharmaceutical: over 100 different commercial rubber formulations
o Stelmi: also, a broad range of commercial rubber formulations
100
Per Material, Supplier and Grade:
what makes up the Impurities Profile?
 Solvent residues (e.g. of Polymerization)
 Polymer residues (e.g. Monomers, Oligomers)
 Catalysts
 Polymer/Rubber Additives
o Antioxidants
o Photostabilizers
o Plasticizers
o Lubricants
o Acid Scavangers
o Pigments/Colorants
o Carifying/Nucleating Agents
o Cross Linking Agents (Rubbers)
o Initiators (Rubbers)
o Accelerators (Rubbers)
 Polymer Additive Degradation Products
 Polymer Degradation Compounds
 Adhesives
101
EXAMPLE FOR 1 GRADE OF POLYOLEFIN
POLYMER EXTRACTABLES: SUM OF
1. INITIAL INGREDIENTS
2. IMPURITIES (e.g. monomers, polymer oligomers, solvent residues, glue

residues, catalyst residues…)
3. REACTION/DEGRADATION PRODUCTS OF POLYMER ADDITIVES
4. DEGRADATION PRODUCTS OF POLYMER

EXAMPLE FOR POLYOLEFINS
antioxidants
1. INITIAL INGREDIENTS O
C2H5 HO
OH O
P
O
NH2
O O
O O
CH3
O
O
H3C OH
C2H5
Stearic acid/stearate OH
O
OH
NC-4/ erucamide
OH CH3
H3C O
Millad
OH
O
HO
O
Glycerol monostearate
O O
O O
O
OH
O
HO
And many others... O CH3
HO
2. IMPURITIES (e.g. monomers, polymer oligomers, solvent residues, catalyst

residues…)
HO OH
O
H3C CH3 OH
H3C O OH
OH
OH CH3 CH3 CH3
O
H3C
OH CH3
H3C O H3C O
Solvent H3C O
CH3
OH
O
OH
residues H3C
CH3
H3C O
CH3
Palmitic acid/palmitate
“Glycerol monostearate”
impurities
Polymer And many others...
oligomers
O O
CH2
OH
O
O
O Irganox 1010 HO
O
O
O CH3
O
HO
O O
OH
OH
O OH

OH
O
HO
O
OH
O
O
HO O O
O O
O O
O
O O
O HO
OH
O
HO
O
HO
O
O O
O O
O O
OH
O
O
HO
OH HO
O
O O
O O
O
OH
O
HO
Irganox 1010
And many others...

4. DEGRADATION PRODUCTS OF POLYMER
O H3C CH3 CH3 CH3 CH3 CH3 CH3
O O CH3
H3C OH H3C H 3C CH3
H OH H CH3 CH3
H3C
CH3
O O H3C CH3 CH3 CH3 CH3
O H3C OH
CH3
H3C OH CH3 H3C H3C CH3
H CH3
O
H3C CH3 O H3C CH3 CH3
H3C OH
OH H3C CH3
H3C H3C CH3
acids aldehydes alcohols ketones Olefin fragments

/ oligomers
Conclusion:
1. The broad diversity of pharma containers, materials, suppliers and
grades, leads to a extremely long list of potential impurities (leachables),
introduced into the drug product
2. The compounds cannot be investigated with 1 analytical technique.

Typically, at least 3 to 5 analytical techniques will need to be combined.
3. Compound Identification is of high importance, therefore the detection

needs to be compound specific (e.g. MS-detection)
• Headspace GC/MS – Volatile Organic Compounds
• GC/MS – Semi-Volatile Organic Compounds
• LC/MS – Non-Volatile Organic Compounds
• ICP – Metals
• IC – Anions
108
Conclusion:
4. For Companies / Labs, only performing E/L-testing, every E/L-project

could turn out into a high level research project (with the need for high
level analytical techniques) because of the lack of materials knowledge
5. For Labs, performing E/L-studies on a routine basis, excessive analytical

costs (associated with high-end analytical procedures) should be avoided
in FIRST PASS testing.
Toxikon: TOX-RAY development
109
What can happen
if the Qualification of the Materials
of the Container/Closure system
has not been performed
properly?
LEACHABLES Profile of a Pre-Filled Syringe
GC/MS Chromatogram of the analysis of the complex drug

solution in a PFS aged for 12 months at 30 °C and 75 % RH.
LEACHABLES Profile of a Pre-Filled Syringe
GC/MS Chromatogram of the analysis of the complex drug

solution in a PFS aged for 12 months at 30 °C and 75 % RH.
Same chromatogram as in
previous slide, but blown up.
36 Leachables!
Sourcing: where do these

compounds come from?
A Novel, Integrated Comprehensive
Approach towards E&L Testing
Integrated Approach for E&L testing
Extractables
• Potential
• Identify Leachables
• Early Risk
TOX- TOX- Assessment
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
• Qualify • Screening
• Source Leachables
Leachables
• Control • Synthesis
• Quantify
E&L ANALYTICAL TECHNIQUES
SAME FOR BOTH

EXTRACTABLE & LEACHABLE STUDIES
HOWEVER:
DIFFERENT FOCUS!!
EXTRACTABLES STUDIES
• INCREASE THE KNOWLEDGE ABOUT THE
COMPOSITION OF THE POLYMER
• FOCUS: IDENTIFICATION OF EXTRACTABLES
• ADDS TO INFORMATION PROVIDED BY RAW MATERIAL

SUPPLIERS OR C/C MANUFACTURERS
• EXTRACTABLES LIST: FOCUS FOR LEACHABLE STUDY
• IN SOME CASES: QUANTITATIVE EXTRACTABLES

STUDIES (e.g. inhalation)
LEACHABLES STUDIES
• TRYING TO ASSESS THE LEACHING BEHAVIOUR

• ASSESS POTENTIAL TOXIC CONSEQUENCES
• FOCUS ON QUANTIFICATION OF “TARGET” COMPOUNDS
KNOWN POLYMER ADDITIVES USED
VALIDATION PACKAGE OF CONTAINER SUPPLIERS
EXTRACTABLES STUDY INFORMATION
TARGETS AFTER Toxicological (ie TOX-RITE) EVALUATION
FOR LVP’S: TARGETS AFTER SCREENING LEACHABLE STUDY
• “SIMULATED USE” CONDITIONS

STORAGE TIME / TEMPERATURE / HUMIDITY
CONDITIONS: SIMILAR TO STABILITY STUDIES
PHARMACEUTICAL FORMULATION AS CONTACT SOLUTION
• VALIDATED METHODS (ICH Q2(R1))
HEADSPACE GC/MS
• EXTRACTABLE STUDIES: Heat the polymer material in a closed vial to

a “polymer process temperature” – avoid thermal degradation
T
Headspace
Volatile organic
compounds
• Desorption of the volatile “impurities” – potential leachables

• Inject the headspace of the vial into a GC/MS
• Similar for solutions:
T Headspace
Volatile organic
compounds
Autosampler
Headspace GC/MS
A b u n d a n c e
S c a n 1 2 8 6 ( 1 3 . 0 0 1 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s
A b u n d a n c e 9 5 .2
2 4 0 0 0 0 0
S c a n 9 6 4 (1 0 . 5 9 2 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s
6 9 . 1
2 2 0 0 0 0 0
A bundanc e
7 5 0 0 0 0
2 0 0 0 0 0 0
S c a n 6 6 8 ( 8 . 3 7 7 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s 7 0 0 0 0 0 A b u n d a n c e
9 3 .1
1 8 0 0 0 0 0 S c a n 2 0 0 9 ( 1 8 . 4 1 1 m in ) : 0 2 J U L 0 0 9 . D \ d a t a . m s
6 5 0 0 0 0 1 3 6 .2
1 5 0 0 0 0 0
600000
6 0 0 0 0 0 1 6 0 0 0 0 0 1 4 0 0 0 0 0
550000 1 3 0 0 0 0 0
6 9 .1
5 5 0 0 0 0
1 4 0 0 0 0 0 9 5 .2
1 2 0 0 0 0 0
5 0 0 0 0 0
500000 1 2 0 0 0 0 0 1 1 0 0 0 0 0
1 2 1 .1
4 5 0 0 0 0 1 0 0 0 0 0 0
450000 4 1 . 2 1 0 0 0 0 0 0
9 0 0 0 0 0
4 0 0 0 0 0
1 0 0 . 1
8 0 0 0 0 0 8 0 0 0 0 0
400000 3 5 0 0 0 0
7 0 0 0 0 0 4 1 .2
6 0 0 0 0 0
3 0 0 0 0 0 6 0 0 0 0 0
350000
1 1 0 .2
4 0 0 0 0 0 5 0 0 0 0 0
2 5 0 0 0 0
4 1 .1 6 7 .1 1 3 6 .2
300000 4 0 0 0 0 0
2 0 0 0 0 0 2 0 0 0 0 0 8 2 .2
3 0 0 0 0 0
250000 5 4 .1 1 2 3 .2 1 5 4 .2
1 5 0 0 0 0 2 0 7 .1
0 2 0 0 0 0 0
7 9 .1 5 5 . 2 8 5 . 1 3 0 4 0 5 0 6 0 7 0 8 0 9 0 1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 1 6 0 1 7 0 1 8 0 1 9 0 2 0 0 2 1 0
1 7 9 .2
m / z --> 1 0 0 0 0 0 1 1 1 .2 1 5 3 .2
1 0 7 .1 1 0 0 0 0 0 2 2 2 .2
200000 1 9 5 .2 2 5 2 .8
0
5 0 0 0 0
4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0
m / z -->
150000 1 1 3 . 1 1 2 9 . 1
1 4 3 . 1 1 9 5 . 0
0
3 0 4 0 5 0 6 0 7 0 8 0 9 0 1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 1 6 0 1 7 0 1 8 0 1 9 0 2 0 0
1 3 6 .2 m / z -->
100000 4 1 .1
6 5 .1
50000
1 5 8 .9 2 0 6 .7
0
30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210
m / z -->
Abundanc e
Sc an 1992 (18.283 min): 02JU L009.D \ data.ms

121.1
Abundance 4500000
S c a n 2 3 5 (5 .1 3 7 m in ): 0 2 J U L 0 0 8 .D \ d a ta .m s (-8 9 ) (-) 4000000

8 6 .0
110000
3500000
100000
90000 3000000 147.1
80000
4 1 .1
2500000
70000
2000000
60000
1500000 91.1
50000
40000 1000000
30000
500000
20000 6 9 .1 59.1
172.1
38.2 203.1 235.1 327.0
10000
0
40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
1 3 2 .9 2 0 7 .1 2 5 3 .1
0 m/ z-->
40 60 80 100 120 140 160 180 200 220 240
m / z -->
Abundance
Scan 3021 (25.982 min): 02JUL009.D\ data.ms

4000000
Abundance
69.1
3500000
S c a n 1 5 1 5 (1 4 . 7 1 4 m in ): 0 2 J U L 0 0 9 . D \ d a t a . m s
7 1 .1
3500000 3000000
2500000
3000000
2000000
2500000
1500000
2000000 41.2
1000000 253.2
1500000 500000
98.1 136.1 183.2 310.3
159.1 224.2 282.3 338.2
0
1000000 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340
4 3 .2
m/ z-->
500000
9 9 .1 1 2 4 .1 1 4 0 .1 1 6 9 .0 2 0 7 .0 2 3 9 .3
0
40 60 80 100 120 140 160 180 200 220 240
m / z -->
Abundance
7 1 .1
3500000
3000000
2500000
2000000
1500000
1000000
4 3 .2
500000
9 9 .1 1 2 4 .1 1 4 0 .1 1 6 9 .0 2 0 7 .0 2 3 9 .3
0
40 60 80 100 120 140 160 180 200 220 240
m / z -->
HEADSPACE GC/MS: Examples of target compounds
AROMATIC COMPOUNDS: e.g. Benzene, Toluene, Ethylbenzene, Xylenes (POLYSTYRENE)

1,3-Bis-(1,1-dimethylethyl)benzene (POLYPROPYLENE)
ALDEHYDES, e.g. Butanal, Pentanal, Hexanal (LDPE, EVOH)

2-Ethylhexanal (PVC)
2-Methyl-2-propanal (PP)
ALCOHOLS: e.g. 2-Ethyl-1-hexanol, 1-Octanol (PVC)

Ethanol (EVOH)
2-Methyl-2-propanol (HDPE, PP)
KETONES: e.g. Acetone, 2-Butanone, 2-Hexanone, 3-Hexanone (PP, LDPE)

Methylethylketone (EVOH)
ACIDS: e.g. Acetic, Propionic, Butanoic, Pentanoic Acid (PP, LDPE)
ESTERS: e.g. Acetic Acid, 2-ethyl ester (PVC, EVA)
ALKANES (PE, PP) ...
The TOX-RAY Database, developed by Toxikon, contains about 650

Volatile Compounds (analytical standards with known Retention Time and
Mass Spectrum) for Rapid Identification
EXTRACTION GC/MS
• Extraction in an appropriate solvent

(e.g.DCM, IPA, Hexane, WFI...)
• Release of semi-volatile “impurities”

– potential leachables
• Inject the extract into a GC/MS REFLUX EXTRACTION
Autosampler
GC/MS
EXTRACTION + GC/MS: Examples of target compounds
POLYMER ADDITIVES: e.g. Butylated Hydroxytoluene

Phthalates
Oleamide
Erucamide
Irgafos 168
Irganox 1076
DEGRADATION PRODUCTS: e.g. 2,6-di-tert-butyl-p-benzoquinone

1,3-di-tert-butylbenzene
Ethylbenzaldehyde
2,4-di-tert-butylphenol
3,5-di-tert-4-hydroxybenzaldehyde
OTHER IMPURITIES: e.g. Crown-ethers, alkylparabens, styrene , tricyclic sesquiterpenes ,

diterpenes, 1.4-Isopropanol acetophenone
MONOMERS &OLIGOMERS: PBT-dimer,

Rubber oligomers,
Polyoxymethylene oligomers,
Acrylonitrile-Butadiene-Styrene
oligomers,
Polyester Oligomers
The TOX-RAY Database, developed by Toxikon Europe, contains about
1550 Semi-Volatile Compounds (analytical standards with known
Retention Time and Mass Spectrum) for Rapid Identification
EXTRACTION UPLC/HRAM: IDENTIFICATION OF
NONVOLATILE COMPOUNDS
• Extraction in an appropriate solvent

(e.g.DCM, IPA, Hexane, WFI)
• Release of non-volatile “impurities” – potential

leachables
• Inject the extract into a UPLC/HRAM High

Resolution Accurate Mass (Orbitrap Technology)
• Can be run in:

 Quantitative analysis
 Screening mode
LC/MS
EXTRACTION + LC/UV/MS: some target compounds
POLYMER ADDITIVES: e.g. BHT OLIGOMERS: e.g. PBT dimer

Irganox 1010 PBT trimer
Irganox 245 PBT tetramer
Irganox 1330 PBT pentamer
Irganox 1076 Caprolactam oligomers
Irganox 3114 ABS oligomers
Hostanox 03
Oleamide
erucamide
EBS
Irgafos 168
Irgafos 126
A.O. 2246
DEGRADATION PRODUCTS: e.g. oxidized Irgafos 168, Irganox 245 degradation products, Irganox 1010
degradation products
NUCLEATING AGENTS: e.g. Millad 3988

NC-4
FATTY ACIDS: Myristic acid, Palmitic acid, Oleic acid, Stearic acid
The TOX-RAY Database, developed by Toxikon Europe, contains about

900 Non-Volatile Compounds (analytical standards with known Retention
Time and Mass Spectrum) for Rapid Identification
TOX-RAY Screener Database A bundanc e
S c a n 6 6 8 ( 8 . 3 7 7 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s
9 3 .1
A b u n d a n c e
7 5 0 0 0 0
7 0 0 0 0 0
6 9 . 1
S c a n 9 6 4 (1 0 . 5 9 2 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s
A b u n d a n c e
2 4 0 0 0 0 0
2 2 0 0 0 0 0
2 0 0 0 0 0 0
S c a n 1 2 8 6 ( 1 3 . 0 0 1 m in ) : 0 2 J U L 0 0 8 . D \ d a t a . m s
9 5 .2
A b u n d a n c e
1 8 0 0 0 0 0 S c a n 2 0 0 9 ( 1 8 . 4 1 1 m in ) : 0 2 J U L 0 0 9 . D \ d a t a . m s
6 5 0 0 0 0 1 3 6 .2
1 5 0 0 0 0 0
600000
6 0 0 0 0 0 1 6 0 0 0 0 0 1 4 0 0 0 0 0
550000 1 3 0 0 0 0 0
6 9 .1
5 5 0 0 0 0
1 4 0 0 0 0 0 9 5 .2
1 2 0 0 0 0 0
5 0 0 0 0 0
500000 1 2 0 0 0 0 0 1 1 0 0 0 0 0
1 2 1 .1
4 5 0 0 0 0 1 0 0 0 0 0 0
450000 4 1 . 2 1 0 0 0 0 0 0
9 0 0 0 0 0
4 0 0 0 0 0
1 0 0 . 1
8 0 0 0 0 0 8 0 0 0 0 0
400000 3 5 0 0 0 0
7 0 0 0 0 0 4 1 .2
6 0 0 0 0 0
3 0 0 0 0 0 6 0 0 0 0 0
350000
1 1 0 .2
4 0 0 0 0 0 5 0 0 0 0 0
2 5 0 0 0 0
4 1 .1 6 7 .1 1 3 6 .2
300000 4 0 0 0 0 0
2 0 0 0 0 0 2 0 0 0 0 0 8 2 .2
3 0 0 0 0 0
250000 5 4 .1 1 2 3 .2 1 5 4 .2
1 5 0 0 0 0 2 0 7 .1
0 2 0 0 0 0 0
7 9 .1 5 5 . 2 8 5 . 1 3 0 4 0 5 0 6 0 7 0 8 0 9 0 1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 1 6 0 1 7 0 1 8 0 1 9 0 2 0 0 2 1 0
1 7 9 .2
m / z --> 1 0 0 0 0 0 1 1 1 .2 1 5 3 .2
1 0 7 .1 1 0 0 0 0 0 2 2 2 .2
200000 1 9 5 .2 2 5 2 .8
0
5 0 0 0 0
4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0
m / z -->
150000 1 1 3 . 1 1 2 9 . 1
1 4 3 . 1 1 9 5 . 0
0
3 0 4 0 5 0 6 0 7 0 8 0 9 0 1 0 0 1 1 0 1 2 0 1 3 0 1 4 0 1 5 0 1 6 0 1 7 0 1 8 0 1 9 0 2 0 0
1 3 6 .2 m / z -->
100000 4 1 .1
6 5 .1
50000
1 5 8 .9 2 0 6 .7
0
30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210
m / z -->
Abundanc e
Sc an 1992 (18.283 min): 02JU L009.D \ data.ms

121.1
Abundance 4500000
S c a n 2 3 5 (5 .1 3 7 m in ): 0 2 J U L 0 0 8 .D \ d a ta .m s (-8 9 ) (-) 4000000

8 6 .0
110000
3500000
100000
90000 3000000 147.1
80000
4 1 .1
2500000
70000
2000000
60000
1500000 91.1
50000
40000 1000000
30000
500000
20000 6 9 .1 59.1
172.1
38.2 203.1 235.1 327.0
10000
0
40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
1 3 2 .9 2 0 7 .1 2 5 3 .1
0 m/ z-->
40 60 80 100 120 140 160 180 200 220 240
m / z -->
Abundance
Scan 3021 (25.982 min): 02JUL009.D\ data.ms

4000000
Abundance
69.1
3500000
7 1 .1
3500000 3000000
2500000
3000000
2000000
2500000
1500000
2000000 41.2
1000000 253.2
1500000 500000
98.1 136.1 183.2 310.3
159.1 224.2 282.3 338.2
0
1000000 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340
4 3 .2
m/ z-->
500000
9 9 .1 1 2 4 .1 1 4 0 .1 1 6 9 .0 2 0 7 .0 2 3 9 .3
0
40 60 80 100 120 140 160 180 200 220 240
m / z -->
Abundance
7 1 .1
3500000
3000000
2500000
2000000
1500000
1000000
4 3 .2
500000
9 9 .1 1 2 4 .1 1 4 0 .1 1 6 9 .0 2 0 7 .0 2 3 9 .3
0
40 60 80 100 120 140 160 180 200 220 240
m / z -->
OTHER ANALYTICAL
TECHNIQUES
 Ion Chromatography (Acetate/Formate; Bromide/Chloride)
 ICP-OES or ICP-MS (Metals)
 Research of Unknowns:
GC-ToF
LC-Q-ToF/MS
FT-MS
 SEM/EDXA
 Microscope-FTIR
EXTRACTABLE STUDIES
DO’S AND DON’TS
CONFIDENTIAL
Extractable studies: do’s and don’ts
STARTING POINT
REGULATORY EVALUATION
 EXTRACTABLES & LEACHABLES STUDIES ARE

NECESSARY!
IDENTIFIED CRITICAL COMPONENT OF CCS
 RUBBER PLUNGER.
DEPENDING UPON THE DESIGN OF E-STUDIES:
1. LOW # of extractables
2. HIGH # of extractables
DO NOT Select the E-study design on # of Compounds
DO select the E-study design on expected conditions

of use
Main Variables: Type of Solvents/Extraction Vehicles

Material-to-solvent ratio
Extraction Times
EXAMPLE
Which EXTRACTION conditions are relevant/worst case
for a plunger in a 1 mL pre-filled syringe, with a 3 year
shelf life at RT (drug product contains 5% Organic)
OPEN QUESTIONS
• Is an extraction ratio of 1 g/10 mL a relevant E-condition?
• Is a 1 hour reflux in Isopropanol a relevant E-condition?
• Is an 8 hour reflux in the DPV a relevant E-condition?
DO NOT ALLOW SURPRISES IN YOUR LEACHABLE / STABILITY STUDIES!!!
E-study: Take worst case conditions compared to “real use”

THE CRITICALITY OF USING THE DP(V) AS A SOLVENT
• Complex DPV: COMPLEX INTERPRETATION OF E-STUDIES!!

Abundance
1.5e+07
DPV-Rubber Extract
1e+07
5000000
-5000000
DPV Blank Extract
-10000000
5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 45.00

Time-->
VARIABLE 1: TYPE OF SOLVENT/EXTRACTION VEHICLES
• Take a Solvent with a similar polarity, but with a higher

extraction propensity than DP
• Optional: Add a solvent with a different polarity

• Confirmation of results
• Avoid surprises in Leachable study!
• Optional: Perform E-study in Drug Product Vehicle

THE CRITICALITY OF USING THE DRUG PRODUCT
(VEHICLE) (DP(V)) AS A SOLVENT
Perform E-study in Drug Product (Vehicle), suggested in:

FDA-Container/Closure Guidance (1999), (eg parenteral/Ophthalmic)
EMEA-Guideline - immediate packaging (2005)

THE CRITICALITY OF SELECTING DP(V) AS SOLVENT
Similar advantages/disadvantages as for WFI:
ADVANTAGE: simulation of extractables behaviour in DP(V): same

extraction propensity!
DISADVANTAGE: Risk of missing the presence of compounds
- Matrix interference of DP(V) (see previous slide)
Risk of misinterpretation of analytical data
- DP(V) Matrix degradant may be misinterpreted as extractable!
Risk of underestimating the concentration of compounds

- Extraction conditions – may potentially be to mild
- Difficult to select the right set of extraction conditions (e.g.
extraction time, temperature!)
EXAMPLE for DP(V) – does 8 hour reflux mimic a 3 year shelf life?
THE CRITICALITY OF SELECTING DP(V) AS SOLVENT
ADVICE when selecting DP(V) as extraction solution:
1. Combine it with organic model solvent (e.g. IPA, DCM, Hexane)

o Minimize the risk of missing the presence of extractables
2. If necessary: Use validated methods, developed for extraction study with DP(V)
as solvent
o Eliminate matrix interference from DP(V) matrix
o Assess DP(V) matrix degradation during extractable study
3. Consider the right set of extraction conditions, relevant for the DP(V) contact
o Extraction time
o Temperature
IDENTIFICATION OF EXTRACTABLES via GC/MS
DIFFERENT LEVELS
IC: Identified Compound

Analytical Standard Available
Retention time (RT) and Mass Spectrum (MS) confirmed: 100%
Abundanc e
S c a n 4 8 7 7 ( 4 0 . 4 7 6 m in ) : 0 4 A U G 0 1 6 . D \ d a t a . m s
8000
4 4 1 .4
unknown
6000
5 7 .1
Abundance 4000
2000 1 4 7 .1
3 0 8 .3 6 4 6 .6
1 9 1 .1 2 3 7 .1 3 8 5 .3
T I C : 2 5 A P R 0 0 9 . D \ D A T A . M S (* ) 1 0 3 .0 5 8 9 .6
0
50 100 150 200 250 300 350 400 450 500 550 600 650
T I C : 0 4 A U G 0 1 6 . D \ d a t a . m s (* ) 4 0 .4 7 8
m / z -->
Abundanc e
2000000
Chromatogram test article # 9 : I rg a f o s 1 6 8

4 4 1 .0
standard
8000
1500000
6000
5 7 .0
4000
1000000 1 6 .7 8 9
2000 1 4 7 .0
3 0 8 .0 6 4 6 .0
1 9 1 .0 2 3 7 .0
1 0 3 .0 3 8 5 .0 5 8 9 .0
0
1 5 .2 0 5 1 8 .7 6 6 2 1 .8 8 1 2 4 .6 6 9
500000 2 7 .1 8 5
50 100 150 200 250 300 350 400 450 500 550 600 650
2 9 .4 7 9 m / z -->
1 0 .8 5 4 1 4 .1960. 80 0 1318 8. 12. 692 2.0474 2 2 2 . 4 6 2 2 5 . 1 8 02 7 . 6 4 3 3 1 .5 8 3
3 3 . 5 2 18 . 2 4 1
5 .1 0 4 1 0 . 9 6 7 1 5 . 3 595811917 8. 51. 098 76. 90602670222.218111.2..56.76948429369236
1 0 . 0 16 12 . 18 3011.10331412..48.4941.12.9545758452.4.405 2 .359742216679265626. 73. 7.00422042 29 9. 4. 8139714. 5 03 83 . 4 34 5
224.474..102
0
fos 168 -5 0 0 0 0 0
-1 0 0 0 0 0 0
Irgafos 168
Chromatogram standard
5 .0 0 1 0 .0 0 1 5 .0 0 2 0 .0 0 2 5 .0 0 3 0 .0 0 3 5 .0 0 4 0 .0 0 4 5 .0 0
T im e - - >
DIFFERENT LEVELS
MPC: Most Probable Compound
Analytical Standard NOT available
Excellent fit with MS-library (>80%)
A b u n d a n c e
S c a n 2 8 5 2 (2 4 .7 1 8 m in ) : 2 4 O C T 0 1 4 . D \ d a t a . m s
2 0 5 .2
9 0 0 0
8 0 0 0
7 0 0 0
MS spectrum of unknown
6 0 0 0
5 7 .2 1 7 5 .2
5 0 0 0
4 0 0 0
3 0 0 0 2 3 2 .3
2 6 1 .2
9 1 .1
2 0 0 0
1 0 9 .1 1 3 5 .2
1 0 0 0 3 9 .1
1 5 7 .2
0
4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0 2 6 0 2 8 0
m / z -->
A b u n d a n c e
O
# 1 0 9 5 0 8 : 7 , 9 - D i- t e r t - b u t y l- 1 - o x a s p ir o ( 4 , 5 ) d e c a - 6 , 9 - d ie n e - 2 , 8 - d io n e
9 0 0 0
5 7 .0
O
8 0 0 0
O
7 0 0 0
6 0 0 0
2 0 5 .0
MS library match
5 0 0 0
4 0 0 0
1 7 5 .0
3 0 0 0
2 0 0 0 9 1 .0
1 0 9 .0 1 3 5 .0 2 3 2 .0
1 0 0 0 2 6 1 .0
1 5 3 .0
0
4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 2 2 0 2 4 0 2 6 0 2 8 0
m / z -->
MPC: 7,9-Di-tert-butyl-1-oxaspiro(4,5)-deca-6,9-diene-2,8-dione
TIC: Tentatively Identified Compound

Analytical Standard NOT available
Lower fit with MS-library (50%-80%): limited structural information
Abundance
S c a n 7 6 5 7 (2 9 . 7 0 6 m in ): 1 8 S E P 0 1 2 . D \ d a t a . m s
4 3 .1 5 7 .1 7 1 .1
8000
MS spectrum of unknown
MS-fit: 72%
T IC : B r a n c h e d H y d r o c a r b o n C 1 2 H 2 6
6000 8 5 .1
Not sure about the

4000
2000 1 1 3 .1 1 2 7 .1
unique identification!
9 9 .1
1 5 5 .1 1 7 0 .0
0
20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170
m / z -->
Abundance
Reported as:
# 3 6 4 5 1 : D e c a n e , 3 , 7 -d im e t h y l-
4 3 .0 5 7 .0
8000 7 1 .0
TIC: Branched alkane C12 6000

MS library match:
4000 8 5 .0
2 9 .0 3,7-dimethyldecane
2000
9 9 .0 1 2 7 .0 1 4 1 .0
1 1 3 .0 1 5 5 .0 1 7 0 .0
0
20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170
m / z -->
IDENTIFICATION OF EXTRACTABLES
DO NOT “OVER RELY” ON MASS SPECTRAL LIBRARIES!!
OPTIMIZE “IDENTIFIED COMPOUND” CATEGORY!!
A UNIQUE IDENTIFICATION IS NECESSARY FOR:
 DOING THE TOX ASSESSMENT ON THE RIGHT COMPOUNDS!
 HAVING THE RIGHT TARGET COMPOUNDS IN YOUR LEACHABLE

STUDY!
EXTRACTABLE STUDIES
Extractable studies
USEFUL DOCUMENTATION PRIOR TO E-STUDY
GENERAL INFORMATION
Product Name, Product N°, Type, Manufacturer, Physical properties…
CERTIFICATES of compendial tests

USP<381>, USP <87>, USP<88>, EP 3.2.9, JP<49>, ISO 8871
INGREDIENTS OF RUBBER
Very useful information, but this will not tell the complete E-story!!
EXTRACTABLES DATA FROM SUPPLIER

Check relevancy of technical info and test conditions
Extractable studies
RUBBER: BASIC INGREDIENTS
ELASTOMER: Base Material
FILLER: Strength, Physical properties (Clay, Talc, Carbonates)
ANTIOXIDANT: Elastomer protection (e.g. BHT, Irganox 1010)
CURING AGENT: Cross Linking (e.g. S, S-donor, Phenol-formaldehyde…)
ACTIVATOR: gives onset of vulcanisation (e.g. ZnO + Stearic Acid)
ACCELERATOR: speeds up vulcanisation (e.g. carbamates, sulfenamides…)
OTHER INGREDIENTS: Pigments, Stabilizers, Plasticizers, Release agents…

Extractable studies
RUBBER EXTRACTABLES: SUM OF
1. INITIAL INGREDIENTS OF THE RUBBER FORMULATION
2. IMPURITIES OF THESE INGREDIENTS

(e.g. residual solvents, oligomers in Elastomer, halides in halobutyl rubber…)
3. REACTION/DEGRADATION PRODUCTS DURING RUBBER PRODUCTION

Extractable studies
IF PROVIDED INFORMATION IS NOT AVAILABLE/SUFFICIENT:
SET-UP AN EXTRACTABLE STUDY
1. DESIGN YOUR E-STUDY, SO THAT IDEALLY:

“LEACHABLES ARE A SUBSET OF EXTRACTABLES”
2. EXTRACTABLE STUDIES: DEFINE MAXIMUM RISK FOR INTENDED USE

IDENTITY OF EXTRACTED COMPOUNDS
CONCENTRATION (RANGE) OF COMPOUNDS
Extractable studies
EXTRACTABLE STUDIES
 Increase KNOWLEDGE About Qualitative Composition
of Polymer/Rubber
 Focus: IDENTIFICATION of Extractables: TOX-RAY!!!
 ADDS TO INFORMATION Provided by either Raw

Material Suppliers or C/C Manufacturers
 Extractables List: FOCUS for Leachable Study
 In Some Cases: QUANTITATIVE Extractable Studies

Extractable studies
Combination of Analytical “BASIC” SCREENING Techniques
• Headspace GC/MS: Volatile Organic Compounds
Monomers, Solvent Residues, Small Polymer (Additive) Degradation Products (e.g. low
MW organic acids, esters, ketones, alcohols aldehydes…), Small Oligomers…
• Extraction GC/MS: SemiVololatile Organic Compounds

Low MW Polymer Additives (Plasticizers, Anti-Oxidants…), Their Degradation Products,
(Halo-)Oligomers, Solvent residues…
• Extraction UPLC/HRAM MS: NonVolatile Polymer Additives

High MW Anti-Oxidants, Slip Agents, Acid Scavengers, S8
• ICP: Metals Analysis

e.g. Inorganic Filler Material can release metals
• Ion Chromatography:
Bromide, Chloride residuals
Extractable studies
VARIABLE 1: TYPE OF SOLVENTS / EXTRACTION VEHICLES
PQRI: combination of POLAR and NON-POLAR solvents

Isopropanol, Hexane, Dichloromethane, WFI
OTHER: acidified WFI
alkaline WFI
saline/buffered WFI
WFI/EtOH-mix (e.g. to mimic Organic content)
Drug Product Vehicle (in FDA-Guidance, PQRI)
Extractable studies
VARIABLE 2: MATERIAL-TO-SOLVENT RATIO - during extraction
DO NOT USE FIXED RATIO’s LIKE:
 2 g / 250 mL (e.g. EP 3.1.3, sample prep for quant analysis of AO’s)

 1 g / 10 mL
RATHER: ADJUST M-t-S RATIO TO THE REAL LIFE RATIO’s
EXAMPLE: for a plunger of a 1 mL Pre-filled Syringe, try to stay as

close to the 1 plunger / 1 mL ratio.
E.g. reflux at a ratio of 1 plunger / 10 mL (fully submerged), and

concentrate 10x afterwards
Extractable studies
VARIABLE 3: EXTRACTION TIMES
Difficult parameter to extrapolate from “real use”
EXAMPLE: for WFI – does 8 hour reflux mimic a 3 year shelf life?
for IPA – does 1 hour reflux reflect a “worst case”?
Alternative Approach:
PQRI: “Determine Asymptotic Extraction Profile”
Extractable studies
Asymptotic Extraction Profile:
PQRI-Example: Test Article: Sulphur Cured Elastomer

Extraction: DCM – Soxhlet
CONCLUSION: Extraction conditions on the ‘plateau’-regime = “MAXIMUM RISK”

Extractable Studies Design
For a PRE-FILLED SYRINGE: Sources of Impurities,
coming from packaging:
 BARREL: Metals (may not be necessary to be studied in EXT Study, if
glass composition is available, direct assessment in LEA study)
Silicone Oil residues may cause protein aggregation
 Rubber Plunger:
 Typically, higher migration when solution is in contact (inverted)
 Migration will be determined by:
 Solubility of leachables in Reconstitution Solution (typically inorganic
aqueous solution (typically low solubility for most non-polar organics)
 Potential Diffusion of Compounds through rubber, into solution
 Temperature
 VOC, SVOC and NVOC may cause a safety issue
 VOC, SVOC, NVOC, Silicone Oil and some Metals may also be Reactive
with DP: also potential Performance & Quality Issue!
 Also, Ions may need to be “checked off”...
152
For a PRE-FILLED SYRINGE: Sources of Impurities,
coming from packaging:
• GLUE for staked needle: Metals (may not be necessary to be studied in EXT
Study, if glass composition is available, direct assessment in LEA study)
• TUNGSTEN Residues: May cause protein aggregation

• NEEDLE SHIELD:
• No Direct Contact between DP and Needle Shield
• HOWEVER: Release of Volatile (VOC) and Semi-Volatile (SVOC)
Compounds from the Needle shield into the content of the PFS is possible!
• VOC and SVOC may also be Reactive with DP : also potential
Performance & Quality Issue!
• Typically No NVOC, Metals and Ions investigation is necessary for a
Needle Shield.
153
EXAMPLE of Potential set-up for an extraction study on the rubber

components of the Pre-Filled Syringe
Plunger Needle shield
TARGET COMPOUNDS ANALYTICAL METHOD
WFI IPA Neat IPA Neat
Volatile Organic Compounds (VOC) Headspace- GC/MS - × × × ×
Semi-Volatile Organic Compounds

GC/MS × × - × -
(SVOC)
Non-Volatile Organic Compounds

LC/MS (APCI+) × × - - -
(NVOC)
Non-Volatile Organic Compounds

LC/MS (APCI-) × × - - -
(NVOC)
Sulfur (S8) LC/UV × × - - -
Elements: Al, Ca, Fe, Mg, Si,

ICP × - - - -
S (total), Ti, Zn
Anions: Br-, Cl- and F IC × - - - -
154
EXAMPLE of Potential set-up for an extraction study on the glass barrel of the
Pre-Filled Syringe for Reconstitution Solution
ANALYTICAL
TARGET COMPOUNDS NH3* WFI DCM mix** Hexane
METHOD
Semi-Volatile Organic Compounds

GC/MS - - X - -
(SVOC) – Glue Residues
TMPTMA (Marker for Glue residue) LC/UV - - - X -
Silicone oil GF-AAS - - - - X
Elements: Al, Ba, B, Ca, Cu, Fe, Mg, Mn, K, S(total),

ICP - X - - -
Na, Ti, W, Zn
Elements: W ICP X - - - -
*: 5 % NH4OH in WFI; **: TFA/MeCN/WFI 0.2/30/70
155
Extractables
• Potential
• VOC • Early Risk
• SVOC
• NVOC
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
Extractable/Leachable Sources
Potential
Inorganic
Compounds
Metals
of concerns
If you don’t look for it, you won’t find it!

157
Volatile Organic Compounds:

» Monomer Residues
» Solvent Residues from Production steps
» residues from polymer treatments (e.g. washing)
» Small Polymer Breakdown products
158
Semi-Volatile Organic Compounds:

» Lubricants
» Plasticizers
» Antioxidants
» Polymer degradation products
» Solvents with an elevated boiling point
159
Non-Volatile Organic Compounds:

» Fillers
» Plasticizers
» Antioxidants
» Polymerization or Hydrogenation Catalysts
» Anti-slip agents
160
Leachable Sources
Inorganic
Metal base complexes or compounds with metals
in their molecular structure
» Fillers
» Pigments
» Catalyst Residues
Potential
Inorganic
Compounds
of concerns
Metals
ICP
161
Extractables
• Potential
• Identify
Leachables
• Early Risk
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
TOX-RITE Evaluations
WHY Integrate (Toxicological) Structure Activity Relationship Assessment
• Extractables studies: long lists of compounds!!
• Extractable reports only give analytical information, but no toxicological

Information is given.
• Toxicological information is often very limited, even for fully identified

Compounds.
• For many compounds toxicological information is even non-existent.

• polymer oligomers
• polymer degradation compounds
• polymer additive degradation compounds
• reaction products (e.g. rubbers)
WHY Integrate (Toxicological) Structure Activity Relationship Assessment
• A basic risk/toxicological assessment on extractables is necessary to

define the target compounds to be monitored during a leachable study.
• lack of accurate toxicological information on the reported compounds

makes these basic risk/tox assessments cumbersome!
HOW are (Toxicological) Derek Nexus Assessments integrated?
• Toxikon has built an E/L specific database (TOX-RAYTM), often detected in

these analytical studies.
• In a first step, all compounds of the TOX-RAYTM database were submitted

to a Cramer Classification. This allowed classifying all compounds into 3
different classes:
Class I: substances of simple chemical structure with known metabolic pathways &
innocuous end products which suggest a low order of oral toxicity
Class II: substances that are intermediate in toxicity
Class III: substances with chemical structures that permit no strong initial impression
of safety & may even suggest a significant toxicity
HOW are (Toxicological) Derek Nexus Assessments integrated?
• Although the Cramer Classification provides value, it cannot be used to

assess:
• genotoxic/mutagenic/carcinogenic compounds
• irritants
• sensitizers
• endocrine disruptors
• In a second step, all compounds of the TOX-RAYTM database were

submitted to a Structure Activity Relationship Assessment (SAR),
using Derek Nexus.
HOW are (Toxicological) Derek Nexus Assessments included?
• The suggested Threshold, per extracted compound, allows to determine
which compounds should be monitored during a leachable study and to
what concentration these compounds should be determined
(i.e. “how low to go” for each target leachable compound).
• This takes into account:
 The Route of administration,
 The total volume of the pharmaceutical container
 The (daily) administration regimen.
• The suggested Threshold will be argued, with reference to the current

relevant guidelines (PQRI OINDP, PQRI PODP,...).
WHAT are the Benefits of the TOX-RITETM approach?
• By integrating the Cramer Classification and Derek

assessments into the analytical extractable reports,
extractable compounds of concern will be identified
immediately.
• This will allow selecting the right target compounds for the
subsequent leachable study from the start of the leachable
study design onwards.
WHAT are the Benefits of the TOX-RITETM approach?
• Resources for toxicological evaluation can be minimized.

• Lead time to completing an E&L study can be minimized.
• TOX-RITETM can reduce/avoid costs for additional

toxicological evaluations
• With this new approach, toxicologists may only need to

“validate” the results of the Cramer Classification/Derek
evaluation and suggested Threshold levels.
Extractables
• Potential
• Identify
Leachables
• Early Risk
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
“Screening Leachable Study”
• As a Step in between an Extractable Study and a “FORMAL” Leachable

study
• Understanding the “Long Term” Interaction between the Container/

Closure System and the Drug Product.
• Screening Methodology to verify:

– The Extractables that have become Leachables
– Unexpected Leachable Compounds
– Allows to narrow down the list of relevant compounds for a leachable study
• With the same techniques as were selected for the Extractables Study
• Easy if some DP is already available in ongoing stability studies (e.g.

Accellerated Conditions)
• For Large Volume Parenterals: Further Narrowing down the list of target
compounds for a Formal Leachable Study
“Screening Leachable Study”
• Feasibility of a Screening Leachable Study depends largely on the

complexity of the DP (the DP should allow screening methodologies)
• Screening Leachable Study is typically “semi-quantitative” in nature (no

validated methods
• Could be performed in a more quantitative way by adding a Method

Suitability Check for certain target compounds.
– To verify if the screeing methodology is capable of picking up certain target
compounds at relevant concentrations in the DP
• If the purpose would be to verify if there are no unexpected leachbles

(leachables that were not detected as extractables):
– A dding ascreening mode study in the Formal leachable study could help to addres
this issue
Extractables
• Potential
• Identify
Leachables
• Early Risk
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
LEACHABLE STUDIES
ANALYTICAL CHALLENGE
 Looking for trace impurities in often very complex matrices!!
 Analytical Methods for Stability Testing are often NOT

suited/optimized for detection of Leachables
o Historically: if no additional peaks (impurities) were found in the

HPLC/UV chromatogram, no leachables
o However: HPLC/UV is optimized for stability/QC testing on API.

HPLC/UV is NOT optimized for the detection of trace impurities
originating from the primary packaging!
LEACHABLE STUDIES
ANALYTICAL CHALLENGE
 Use dedicated and optimized analytical tools for

leachables analysis, e.g.:
o Headspace GC/MS: volatile compounds
o Sample prep. + GC/MS or GC/QQQ: semi-volatile compounds
o Sample prep. + LC/MS or LC/QQQ: non-volatile compounds
o ICP: metals
o IC: ions
 PQRI- threshold considerations for leachables

LEACHABLE STUDIES
 “Simulated Use” Conditions

Storage Time / Temperature / Humidity
Conditions: Similar to Stability Studies
Pharmaceutical Formulation As Contact Solution
 Quantification of “Target” Compounds

Extractables study information
Targets, Identified after TOX-RITE evaluation
Screening Leachable Information
Composition of Raw Materials/Compounds
Validation Binder of Container/Closure Manufacturer
LEACHABLE STUDIES
 Validated Methods (ICH Q2(R1))
• Specificity - Identification
• Range
• Linearity of Method
• Extraction Yields (when applicable)
• Detection Limit
• Quantification Limit
• Accuracy in low, mid and high concentration range
• Precision in low, mid and high concentration range
Other: Intermediate Precision, Robustness...

Leachables Study Design
Pre-Filled Syringes have LONG TERM EXPOSURE to either

the Drug Product
FULL LEACHABLE STUDY

o Long Term Ageing Conditions
o Accelerated Ageing Conditions can be considered, in support of LT Ageing
o Monitoring Concentrations of target compounds from EXT study, after an
initial toxicological/risk assessment (if using a threshold approach, see part 6)
o At different time points
o Quantitative Methods (Validation) to quantify the compounds in DP
o Screening Methods (semi-quantitative), to pick up unexpected leachables
178
EXAMPLE OF ANALYTICAL PACKAGE FOR A FULL
LEACHABLE STUDY
TARGET COMPOUNDS ANALYTICAL METHOD

VALIDATED METHOD
Headspace GC/MS
Volatile Organic Compounds (VOC) SCREENING
VALIDATED METHOD
GC/MS
Semi-Volatile Organic Compounds (SVOC) SCREENING
VALIDATED METHOD
LC/MS
Non-Volatile Organic Compounds (NVOC) SCREENING
Element Analysis ICP
Anions: fluoride, chloride, and bromide IC
Sulfur (S8) LC/UV
179
EXAMPLE OF FULL TIME TABLE LEACHABLE STUDY
Storage Time (Months)

Type of Solution
0 3 6 12 24
Drug Product in Rubber Sealed Vials (Test Item) at 5 ± 3 °C × × × × ×
Drug Product in Inert Containers (Blank) at 5 ± 3 °C × × × × ×
Drug Product in Rubber Sealed Vials (Test Item) at 25 ± 3 °C - × × - -
Drug Product in Inert Containers (Blank) at 25 ± 3 °C - × × - -
× = sampling time point
180
EXAMPLE OF FULL EXTRACTABLE&LEACHABLE STUDY
t = -8 t=0 t=3 t=6 t = 12 t = 24
EXT VAL Leachables Study
Administrative
Evaluation Evaluation
lead time
Total = 34 months
181
What to do if
some of the target compounds
are not commercially available
for quantification?
Extractables
• Potential
• Identify
Leachables
• Early Risk
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
SYNTHESIS AND ISOLATION OF COMPOUNDS FOR
1. Identification/Qualification of “unknown” extractables
2. Quantifying these compounds as a leachable

– For Compounds of Concern, often low levels need to be verified
– Having access to the Analytical Standards allows us to develop, optimize
& validate the methods for these compounds at the appropriate levels
3. In certain cases perform Toxicological Tests

– AMES test
– (Rat) Accute Toxicity Study
– Cytotoxicity
HALOGENATED RUBBER OLIGOMERS: COMPOUNDS
OF HIGH CONCERN IN PARENTERAL APPLICATIONS
Synthesizing compounds, focus for 2012, some examples:

CH2 H3C CH3 CH3
CH2 CH3
CH3
H3C CH3 H3C
H3 C CH3 H3C CH3
H3C CH3
CH2 H3C CH3 CH3

CH2
Cl
Br H3C CH3 CH3 CH2 CH3
Br
H3C CH3 CH2 CH3
H3C CH3
H3 C CH3 Cl H3C CH3
H3C CH3
H3C CH3
H3C CH3
H3C CH3
o Halobutyl rubbers are used in many Parenteral applications (Vials PFS, Needle
shields....)
o Oligomers are a typical material impurity of these rubbers, as Extractable or
Leachable
185
CONFIDENTIAL
C13H23Br/ C13H23Cl and C21H39Br/ C21H39Cl Oligomers
• Considered as
• HALOGENATED Cyclic Aliphatic Hydrobarbon compounds (Allyl Halide)
• Alkylating Agents
• One double bond
• Structure Activity Relationship (SAR) Assessment:
CARCINOGENICITY IN HUMANS IS PLAUSIBLE
• As no experimental data / Literature data is known about the toxicity of these

compounds, a lot of Pharma companies:
• Rely on the result of a SAR assessment to perform a tox evaluation
• Conclude that these compounds are of High Concern
For potential Mutagenic/Carcinogenic compounds:
SCT: 0.15 µg/day (PQRI OINDP)
TTC: 1.5 µg/day (PQRI-PODP; ICH guideline on Genotoxic Impurities)
Low SCT/TTC levels for the Halogenated Oligomers mean:

 Low associated AET levels
 High level of method optimization to obtain these levels (certainly with LVP)
 e.g. SIM mode for GC/MS
 Can only be performed with appropriate analytical standards with known purity
– Method Selectivity
– Accuracy
– Sensitivity
– Precision
187
Observed Reactivity of C13H23Br and C21H39Br
(as alkyating agents) with peptides, proteins, and nucleic acids
O
Example of Lyophized H
N
HN CH2
peptide
Lys OBSERVED & POTENTIAL O
H
N
O
O
H
Reactivity with Peptides (lyo) NH
CH2
H3C CH3
O S
Cys N
HN H3C CH3
O NH2 CH2 H3C CH3
O HN
N H3C CH3
NH SH nucleophilic
O side chain
CH2 attack H3C CH3
N
His Br H3C CH3
Peptide NH
H3C CH3
H3C CH3
O CH3
With different nucleophilic groups H3C
O N
O NH
P Nucleophilic “N7” attack is also
O
O N N NH2 "N7" attack H3C O SN2 reaction mechanism in
O CH3
O
anti-cancer drug Busulfan
O H2C N
NH
DNA O Guanosine Most nucleophilic site P
O O
P O N N NH2
O O
O
O
O
P
O
O
188
Observed Reactivity of C13H23Br and C21H39Br
(as alkyating agents) with peptides, proteins, and nucleic acids
O
H
N
HN CH2
Example of Lyophized OBSERVED & POTENTIAL H
Lys O N
peptide Reactivity with Peptides (lyo) O
O CH2
H NH S H3C CH3
N O
Cys HN H3C CH3
O NH2 CH2 H3C CH3
O HN
N H3C CH3
NH SH nucleophilic
O side chain
CH2 attack H3C CH3
N
His Br H3C CH3
Peptide NH
H3C CH3
With different nucleophilic groups H3C CH3
O CH3
H3C
O N Nucleophilic “N7” attack is also
O NH
P SN2 reaction mechanism in
O anti-cancer drug Busulfan
O N N NH2 "N7" attack H3C O
O CH3
Most nucleophilic site O H2C N
DNA DNA O
O Guanosine
O
O
P
O N
NH
P N NH2
O
O
POTENTIAL O
Reactivity with
O
DNA O
P
O
O
189
o Toxikon is the Only Lab worldwide, having access to these
halogenated oligomers, through their TOXYNTH synthesis/isolation
programs
o By Obtaining the pure Oligomer Compounds, Toxikon was/is able to
 Confirm Identity of these species
 Fully Identify these compounds in “First Pass” Experiments
 Quantify these compounds as a leachable
o The halogenated Oligomers are of high Toxicological Concern

o Access to analytical standards will allow us to develop sensitive
analytical methods to monitor the concentration of these compounds at
very low levels.
o Performing an AMES test on obtained Halogenated Oligomers
TOXYNTH: DEGRADATION OF ADDITIVES
Degradation Study of Polymer Additives:
1. Understand the Degradation Mechanism
2. Identify Degradation Compounds + Include in TOX-RAY

• GC/MS
• Derivatization GC/MS + Derivatisation GC-ToF
• LC-Orbitrap (Accurate Mass)
3. Can we Isolate certain degradation compounds and use

them as standards?
191
CONFIDENTIAL
DEGRADATION OF POLYMER ADDITIVES
UV Ozone
Water
Compound THF H2O2
Solid Solution Reflux
Solid Solution
Irgafos126 x x x x x
Irgafos168 x x x x x x
Irgafos PEPQ x x x x x x
Irganox245 x x x x x x
Irganox1010 x x x x x x x
Irganox1076 x x x x x
Irganox1098 x
Irganox1330 x x x x x x
Irganox3114 x
AntiOxidant2246 x
Hostanox03 x
192
Extractables
• Potential
• Early Risk
RAYTM RITETM
Toxicology TOX-
ASSESSMENT SYNTHTM
Leachables
• Quantify
Analytical Methods for E&L Determination
“FIRST PASS” Testing for Extractable Studies

Try to Fully Characterize the Extraction Profile of material, using:
Standard Analytical Equipment, e.g.

1. Headspace GC/MS
2. GC/MS
3. LC/MS
4. ICP
5. Ion Chromatography
Optimized Procedures & Procotols

» Leverage Compound Database (ie:TOX-RAY) for First Pass Analytical
Techniques built on expertise & high volume of studies  allows high level ID
in First Pass
Dedicated Equipment to Standardized Methods
Efficiency/Cost effectiveness is Key in FIRST PASS Experiments

“SECOND PASS” Testing

Utilize Specialized Analytical Equipment for further identification of any
unknown compounds
High End Analytical Equipment, e.g.

1. LC/MS Orbitrap
2. GC-ToF (often in combination with derivatization)
3. FT-MS
4. NMR
5. SEM-EDXA
Project Related Second Pass Testing

» Evaluate the need to identify unknowns on a case by case basis
» Is it Toxicologically & Physiologically Relevant?
Note: Be sure to collect data and further develop database from

2nd Pass testing to improve subsequent 1st Pass testing
ACCELERATED LEACHABLES testing

In some cases: a SIMULATION study
» As a step in between an Extraction Study and a “Formal Leachable” Study
» Understanding the real risk of which extractabe compounds may become a

leachable
» Account for unexpected leachables and for secondary leachables
» Using screening methodologies, not optimized for the specific matrix.
» Either as qualitative/semi-quantitative Screening (not for submission) or

with more quantitative methods
LEACHABLES testing
Select Targets, based upon
» Extraction Studies
» Accelerated Leachable Studies
» TOX-RITE information or other
Toxicological Assessment information
Validate the methods in Matrix for Adequate Quantitation
Factors that could impact the depth of a validation:

» The dosing regimen (frequency, volume)
» The complexity of the drug product composition
» The toxicity of compounds
» Regulatory requirements, Sponsor requirements
Additional Screening is advised!
Accelerated Leachable study (e.g. with screening methods):
Verify the presence of “unexpected leachables”
Kinetics of Extraction Extraction Accelerated Real time/temp

Leachable St. Leachable St.
H2O DCM or IPA e.g. 6 Mo, 40°C e.g. 3 y at 25°C
e.g. 8h reflux e.g. 8h reflux
EXTRACTION SLOW – FAST – Enhanced SLOW, but long
Incomplete complete Diffusion controlled term contact!
leaching is T-dependent
no swelling/enhanced Enhanced Diffusion
D = D0 exp(-E/RT)
diffusion Almost Asymptotic
MATERIAL Slightly enhanced Very Slightly Slightly enhanced SLOW, but

DEGRADATION ASTM 1980: reflux at enhanced ASTM 1980: evaluated over
100°C/8h: 60d at RT 6 Mo ageing at 40°C ≡
ASTM 1980: (IPA) LONG period!
Even if they will be formed, 17 Mo at 25°C
reflux at 80°C/8h:
will they come out? (e.g. 3y)
15d at RT
REACTION Slightly enhanced Not relevant! Enhanced, SLOW, but

KINETICS Low [extr]init will limit the k = k0 exp(-Ea/RT) evaluated over
formation of reaction Ea: Activation Energy,
• Dissolved O2 in H2O LONG period!
comp. (i.e. for slow reaction dependent
• Hydrolysis (H2O)
reactions) (e.g. 3y)
• Reaction with DP and
(Pseudo) first order
leachates/materials
kinetics

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Extractables & Leachables

• Ensuring the safety and compatibility of

Pharmaceutical Medical Device

Center for Biologics

Center for Food Safety

FDA oversight for medical devices authorized by 1976 Medical Device

Per Chapter 2, §201(h) of Food, Drug, and Cosmetic Act, as amended:

• “A medical device is an instrument, apparatus, implement, machine,

• Intended to: - Diagnose diseases/conditions;

• Does not achieve primary purpose through chemical action in or on the

• Is not dependent upon being metabolized for its primary purpose.

Primary characteristics that distinguish medical devices from drugs.

Chemicals that can migrate from CCS, devices and

The Submitter / Marketer of the Drug or Device

• New Drug or Device Approvals!!!

…and Sustainable Regulatory Compliance

• Will they get the new drug or device approvals?

• A Pharmaceutical Company Submits an NDA

• Was it a good Decision?

• Must Balance Risk vs. Reward

• What is the composition of my component?

An understanding of the chemical and

• Solubility of Chemicals in the Polymer

A polymer is a large macromolecule

• Linear or Branched Framework

Elastomer Thermo set

Physical mixture of two or more different types of polymers

To blend different polymers is an easy way to improve certain

Examples: PP-Copo, ABS, POM,…

Monomer (Particle) A Monomer (Particle) B

Structural Regularity with Little Branching

Hydrogen bonds: PA, POM

Each hydrogen bond has a contribution to the stability of the polymer

LD-PE Density: 0.915 - 0.940m [g/cm3]

Density: 0.940 - 0.965 [g/cm3]

Density of 0.907 [g/cm3]; Crystallinity up to 70%, Mp: 160°C - 208°C;

atactic n most transparent

• Additives are Ingredients Incorporated into the

– Modifiers – Change or Improve Polymers Performance

Used in Both Plastic and Rubber

Other Properties Phosphites consume

R-H Melt Processing

CH2OCH3 CH2OCH3 H3C OH CH3

A controlled incompatibility in the polymer

• Friction between surfaces of two films

• Migration of slip agents towards the surface

• Controlled incompatibility with the polymer

Possible change in leachables profile!

Color can be very important in the marketing of a product.

• Monomers or Oligomers from Incomplete Polymerization

Rubber Curing Agents 9

• Special Case Compounds 6

Monomer Polymer Manufacturing Masterbatchers End Products Pharma

Process Polymerization Antioxidants Light

Upstream Change Control Issues

Greater Sales Volumes to Other Markets

Polymer Manufacturing Masterbatchers

Can Drive Upstream Changes