This document discusses extractables and leachables from pre-filled syringes and their potential impact on biologic drug products. It defines extractables and leachables, outlines analytical techniques used to detect them, and provides examples of leachables detected from syringes that caused protein aggregation or covalent bonding. These included residual nylon, tungsten oxides, acrylate adhesives, and silicone oil. The document emphasizes that pre-filled syringes can contain various extractables and leachables from multiple sources, and that early collaboration with suppliers is important to develop a robust understanding and mitigate risks to drug product safety and quality.

1. Pre-filled Syringe
Extractables/Leachables and
Impact on Parenterals
Yasser Nashed-Samuel, PhD
Amgen
Process Development
5th Annual PFS Summit
June 23rd–25th 2015
San Diego

2. 2
Acknowledgements
 Hans Lee
 Kiyoshi Fujimori
 Mike Ronk
 Jian Liu
 Yanxin Luo
 Alejandra Gallegos
 Pavel Bondarenko
 Ron Forster
 Zai-Qing Wen
 Gianni Torraca

3. 3
Outline
 Definitions
 Extractable/Leachable
– Approach
– Impact on safety
– Examples from prefilled syringes
 Summary

4. 4
Extractables and Leachables definitions
• Extractables - chemicals that migrate from the
product-contact material (container) into a
solvent at elevated temperatures.
(Accelerated Condition)
• Leachables - chemicals that migrate from the
container into a formulated drug during
normal storage/usage conditions.
(Normal Condition)
Solvent
Drug
4 °C
Heat

5. 5
Exractables correlation to Leachables
 Relationship between Extractables and Leachables
– L and E can be the same compound.
– L can be a subset of E (derivative, degraded,
oxidative product)
– L/E can react with the other L/E, formulation, or
product
Extractables
Leachables

6. 6
Extractable vs Leachable Analysis
Extractables
(accelerated, heat)
Leachables
(normal, 4 °C)
Matrix
(Solvent)
Simple
(Drug Product)
Complex
Interference Minimal Yes
Analysis Less complex Complex
Storage/Contact
Time
Days Year(s)
Product
independent
Yes No
 Extractable samples are more practical for analysis
and can be predictive for Leachables

7. 7
Typical Analytical Techniques for
Extractables/Leachables
Technique Used to Detect Sensitivity
ICP/MS Inorganic elements (W, Al, Fe, etc.) ppb
GC/FID
Volatile organic molecules
(Boiling point < 300 °C)
ppm
GC/MS
Volatile organic molecules
(Boiling point < 300 °C)
ppm
HPLC UV absorbing molecules ppm
LC/MS
Organic small molecules (polar,
containing hetero atoms O, N, etc.)
ppb-ppm
LC/ELSD Universal detector ppm
NMR Organic compounds ≥ ppm
Goal: Identify and quantitate extractables/leachables

8. 8
Biologics (Injectables): Highest Risk Category
Guidance for Industry-Container Closure Systems for Packaging Human Drugs and Biologics (1999)

9. 9
Manufacturing Process of Biologics
Cell Culture Syringe, Vial,
Device, IV
Leachable Toxicology Risk
(Proximity to Drug Product)
All materials can leach: Polymers, rubber, steel,
glass, connectors, glue, etc..

10. 10
Extractable/Leachable Potential Impact
 Patient Safety
– Toxicity
 Container
– Functionality and Qualification
 Protein/Drug (Product Quality)1-5
– Aggregation / Particle formation (Tungsten)
– Covalent bonding (Acrylates)
– Oxidation (Metals)
 Cell Culture Yield6 (Antioxidant) 1. Am. Pharm. Rev. Jan 2011, p 74.
2. J. Pharm. Sci. 2009, 98(12), p 4695.
3. J. Pharm. Sci. 2009, 98(6), p 3167.
4. PDA J Pharm Sci Tech 2013, 64 (1) p 11.
5. PDA J Pharm Sci Tech 2010, 64 (3) p 242.
6. PDA J Pharm Sci Tech 2013, 67(2) p123.

11. 11
Proteins in Glass Prefilled Syringes (PFS)
 Advantages of PFS relative to Vials/IV
– Small volume
– Autoinjectors (ease of use)
– Safety
– Accuracy
– Reduced overfill
 Some PFS challenges
– High protein concentrations
– Formulation
– Glass breakage / delamination
– Lubrication / Protein particulation
– Storage time > 1 year
– Increase risk for Leachables

12. 12
Prefilled syringe: Sources of Leachables
Barrel
Stopper
Needle Shield
Needle
Adhesives
Lubricant
Residual from
Manuf. Tools
(Tungsten, nylon)
Sterilization:
Ethylene oxide
Gamma
E-beam
Stopper
coating

13. 13
Case 1: Black Particle in Filled Syringes
Nashed-Samuel, et. al. PDA J Pharm Sci Tech 2010, 64 p 242.
LCMS
Residual Nylon
Residual Nylon
Nylon pins used on assembly line

14. 14
Case 2: Tungsten Oxides and Protein
Aggregation
Observed
Protein Aggregation
in PFS
Detected Leachable:
Tungsten
0
100
200
300
< 0.01 0.1 0.2 0.3 0.4 0.5 0.6 0.7 > 800
Frequency
W, ppm
Tungsten in 8 Lots (500 New Syringes)
Jiang,Y, Nashed-Samuel, Y, et al. J. Pharm. Sci. 2009, 98(12), p 4695.
Lee, H, Nashed-Samuel, Y, et al. PDA Nov 2007 Extractable forum.
Analyses
Tungsten is in all glass syringes
Sensitivity
Visible
Particles
Spike Tungsten
oxide, ppm
Aggregation, %
(SEC)
YES 8.3 1.3
YES 1.0 0.3
YES 1.8 0.2
YES 1.3 0.2
YES 1.3 0.2
YES 1.1 0.1
NO 0.6 Not Detected
NO 0.4 Not Detected
NO 0.01 Not Detected
NO 0.01 Not Detected

15. 15
Residual Tungsten in Glass Syringes
After formation Glued-in needleTungsten pin in
barrel
 Tungsten pin (tool) used to form the needle channel at
~ 1,200 C

16. 16
Tungsten Oxides in Syringes
--
Tungsten oxides
Tungsten oxides
Metallic needle
> 800 C
2WO3
> 150 C
2W + 3O2 Sublimes

17. 17
Case 3: Adhesives from Prototype Syringe
Extractable* Structure
A Acrylic acid
B Methacrylic acid
C
2-hydroxypropyl
methacrylate
D
1-Hydroxy-2-propyl
methacrylate
E
Tetrahydrofurfuryl
methacrylate
O
HO
O
HO
O
O
HO
O
O
HO
O
O
O
*concentration: ng scale (ppb)
Reactive

18. 18
Michael Addition (Lysine)
Uchida K. Amino Acids 2003, 25, 249-257
Michael Donor
Michael acceptor
(Acrylic Acid)

19. 19
Acrylic Acid binds to Protein: Modification
Percentage
Peptide 45 ºC 37 ºC
VDIKR 0.250 % 0.020 %
DIVMTQTPLSSPVTLGQPASISCR 0.028 % 0.006 %
VVSVLTVVHQDWLNGK 0.019 % 0.003 %
WQQGNVFSCSVMHEALHNHYTQK 0.230 % 0.040 %
Leachables can bind to proteins
Liu, D., Nashed-Samuel, Y, et al. PDA J. Pharm. Sci. Tech 2012, 66(1), p 12.

20. 20
Silicone Oil Impact on Protein
 Device impact
– Silicone oil is essential for syringe lubrication
– Syringe stalling and incomplete drug delivery
 Protein impact
– Silicone oil can cause/induce protein aggregation
and visible/sub-visible particle formation
– Intermolecular interactions between silicone oil
and protein
Nashed-Samuel, Y. et al. Am. Pharm. Rev. Jan 2011, p 74.
Thirumangalathu, R. et al. J. Pharm. Sci. 2010, 98(9), 3167-3181

21. 21
Silicone oil movement (4-12 months)
Silicone Oil: Movement
1 3 5 7
Siliconeoil
UP-4 month
Down-4
Down-8
Down-12
4D 8D
12D
4U
 Gravity moves silicone oil within the syringe.
 Syringe storage position and time will influence
silicone oil distribution.
n=10
UP Down
Normal
Storage position
Gradient
Nashed-Samuel, Y. et al. Am. Pharm. Rev. Jan 2011, p 74.

22. 22
Syringe Extractable/Leachable Profile vary by
Suppliers
4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00
Supplier A syringe1 2 3
4
5
 Five compounds observed in supplier A
syringes by GCMS
Supplier B syringe
0
200000
400000
600000
800000
1000000
1200000
1400000
1600000
1800000
2000000
2200000
Abundance

23. 23
Summary
 Prefilled syringes have Extractables/Leachables
– Many sources (tools, stopper, adhesive, etc.)
 Leachables can impact biologic drug products
 Proper supplier controls and formulation design
mitigates potential Leachable impact to product
resulting in robust safe systems.
Partnering early with Suppliers, developing robust understanding of extractables/leachables
during commercialization is critical for success!