EXTRACTABLES & LEACHABLES

Standard Protocols in Single Uses

TechnologyUSP and BPOG Approaches; a discussion in brief

Dr. Andreas Nixdorf

SGS Life Sciences

2017 CPHI Frankfurt

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RISK FACTORS ASSOCIATED WITH E&L

General quality consideration with leachables

Product stability

– Aggregation, increase in particulates, …

Interfere with analytical methods or diagnostic tests

Negatively impact on process performance

– cell growth, rate of drug release, drug solubility, pH,

product yield …

Product safety (toxicological risk)

The leachable may be toxic and poses a health risk to

the consumer

Efficacy consideration

The leachable interacts with the drug product

formulation or API so as to alter its stability and potency

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THE CHALLENGES OF END USER

Functionality

by compatibility

Warning letters

from authorities

Regulatory

requirements

End User

Patient

Safety

Product

Impacts

Economic

Impacts

Regulatory

acceptance

Suppliers data

(complete?)

Supply

Chain

Risk Mitigation

Input

Su

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Qu

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interpretation

Ke

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LITERATURE SOURCES – STANDARD

PROTOCOL DISCUSSIONS

Some key publications

PDA, Application of Single-Use Systems in

Pharmaceutical Manufacturing, Technical Report No. 66

Parenteral Drug Association, Inc. 2014.

Biophorum operations Group (BPOG): Best Practices

guide for evaluating Leachables risk from polymeric

Single Use Sytems used in biopharmaceutical

manufacturing. (2017)

Ding W, Madsen G, Mahajan E, O’Connor S, Wong K,

Standardized Extractable Testing Protocol for Single-

Use Systems in Biomanufacturing, Pharmaceutical

Engineering, 2014; 34(6), 1–11.

Drafted USP <1665> and <665> USP forum (2017)

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If extractables testing data provided by a supplier are not

sufficient, the end –user has to perform own studies;

these results in the same components being tested multiple

times and delay in application of SUS in biomanufacturing.

It would ensure, that a comprehensive and consistent set of

extractables testing data are readily available to the end-

users.

Assures comparability of chemical profiles from SUS

components from same functionality but delivered from

different vendors.

It would assist SUS suppliers in more efficiently selecting

materials in line with end-user needs.

“PROS” OF THE USE OF STANDAD

PROTOCOLS

BPOG „Standardized Extractables Testing Protocol for SUS in Biomanufacturing, Pharm. Eng. 2014.

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Classify your disposables in different risk classes:

A: Baseline assessment

B: Expanded Baseline assessment

C: Full Testing assessment

Please compare with risk assessment procedure as

proposed by BPOG group:

Biophorum operations Group (BPOG): Best Practices guide

for evaluating Leachables risk from polymeric Single Use

Sytems used in biopharmaceutical manufacturing. (2017)

USP <1665/665> - RISK BASED APPROACH

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ESTABLISH LEVEL A OF CHARACTIZATION

Level A (Baseline assessment):

The requirement is that all materials of construction must comply

with USP <661.1>:

• Identity;

• Physicochemical characteristics;

• Biological reactivity test, USP <87>;

• Additives (by proper reference to 21 CFR Indirect Food

Additives Used in Food Contact Substances);

• Levels of extractable metals.

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ESTABLISH LEVEL B OF CHARACTERIZATION

Level B (Expanded Baseline assessment):

One requirement is that all materials of construction must comply

with USP <661.1> and,

• The materials of construction must meet the requirements for

Plastic Class VI designation, In Vivo <88>;

• Additives by “proper” testing (list of containing additives and

levels).

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FOOD CERTIFICATE INFORMATION

Example (Polyurethane) of Food “Proper Testing List” (EU):

CAS Substance Restriction

822-06-0 Hexamethylene diisocyanate QM*(T) = 1 mg/kg

584-84-9 2,4-tolylene diisocyanate QM*(T) = 1 mg/kg

693-36-7 Octadecyl-3-(3,5-di-tert-butyl-4-

hydroxyphenyl) propionate

SML = 6 mg/kg

….. ….. ……

*QM max. conc. of residual in the plastic.

** SML restriction of specific migration limit.

Leaching Residue Potential Hazard

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ESTABLISH LEVEL C OF CHARACTERIZATION

Level C (Full Testing assessment):

One requirement is that all materials of construction must comply

with USP <661.1> and specification for Level B met;

• Materials extractable profile has fully established relevant to

situation.

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HOW MANY STANDARDIZATION WOULD BE

USEFUL? STANDARDIZED EXTRACTABLES

TESTING USP <665>

Components Extraction

(solutions

C1, C2, C3)

Extraction at

40°C

Extraction Duration

1 day 7 days 21 days

Storage container X X – – X

Mixing bag X X X X –

Bioreactor bag X X – – X

Tubing connector and

disconnector

X X – – –

Aseptic/sterile connector and

disconnector

X X – – –

Sensor/valve X X X – –

Molded parts of mixers X X X – –

Polymer pump surfaces X X X – –

Tubing X X – – X

Gasket, O-ring X X X – –

Sterilizating filter X X X – –

Process filter X X – X –

Tangential flow filtration X X X – –

Chromatographic column X X X – –

Filling needle X X X – –

C1: acidic extraction at pH 3; C2: alkaline extraction at pH 10; organic extraction 1/1 ethanol/water (v/v)

Short term (Extraction?) Long term (Use Study?)

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BPOG

Extraction temperature 25°C (=< 30 min contact); 40°C

Extraction time depending on SUS component: 24h, 7d, 21d, 70d

Material to surface ratio 6 cm2/mL

USP

Extraction temperature: 40°C

Extraction time depending on SUS component: 24h, 7d, 21d

Material to surface ratio 6 cm2/mL

Solvents:

BPOG: 50% ethanol, 1%PS-80, 0.1M H3PO4, 0.5N NaOH,

5M NaCl, WFI

USP: 50% ethanol, HCl/NaCl at pH 3, Phosphate buffer pH 10

DIFFERENT PROTOCOLS – EXTRACTION

CONDITIONS

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In general, standard extractable protocols do not take into

account the different chemical nature of polymers or coated

materials.

Not all polymeric materials will show chemical resistance

under certain conditions.

The chemical profiles may be overestimated, that could lead

to a disqualification of a beneficial material.

By protocol multiple orthogonal methods are used to

chemically profile materials extractables.

Particular critical substances may require target analysis,

thus they run out of analytical window by using screening

methods.

“CONS” OF THE USE OF STANDAD

PROTOCOLS

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INADEQUATE CHEMICAL RESISTANCE –

SANITIZATION WITH 1M NaOH

600 cm2 / 150 cm Pt cured Silicone

Tubes

Simulated use study: 40°C&50°C

7d, 1M NaOH

TOC: 122 – 330 µg/cm2;

73 – 198 mg/150 cm

ICP-MS: Si element; saturated

detector after 1:1000 dilution.

Delamination of tube inner surface

(SEM: scanning electron microscope).

High risk of particle formation.

Oligomeric substances.

Degradation mechanism

SEM image of damaged area.

Delamination flakes

Mosammad Shamsun Nahar*, Jing Zhang „ Analysis of

Damaged Silicon Rubber Hose”, American Journal of

Analytical Chemistry, 2011, 2, 363-370

EXAMPLE

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INADEQUATE RESISTANCE - AQUEOUS

ALCOHOL SOLVENTS

Virus filter with PES membrane

Exaggerated Extraction: PBS

buffer pH 7.4 + 25% (v/v) iPrOH,

28d 40°C

Amount of embedded membrane

polymer found: > 350 mg/filter!

Under real use conditions only

minor

quantities are released below

toxicological relevant limit.

The filter membrane was

demonstrated incompatible to

aqueous alcohol solutions.

ESI + MS (QToF) Ion Trace

EXAMPLE

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INADEQUATE RESISTANCE - PARTICLE

FILTER SYSTEM - 50% ETHANOL

Extraction conditions: 50% ethanol at 40°C for 48h:EXAMPLE

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 5 0 . 0 0

2 0 0 0 0

4 0 0 0 0

6 0 0 0 0

8 0 0 0 0

1 0 0 0 0 0

1 2 0 0 0 0

1 4 0 0 0 0

1 6 0 0 0 0

1 8 0 0 0 0

2 0 0 0 0 0

2 2 0 0 0 0

2 4 0 0 0 0

2 6 0 0 0 0

2 8 0 0 0 0

3 0 0 0 0 0

3 2 0 0 0 0

3 4 0 0 0 0

T i m e - - >

A b u n d a n c e

S i g n a l : S c r e e n i n g _ E x t r a c t a b l e s _ 4 0 7 6 4 4 0 _ 1 7 0 6 2 6 _ 2 4 . D \ F I D 1 A . c h

Advanced disintegration of polymeric material: high amounts of low mass oligomers.

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CUSTOMIZE REALISTIC EXTRACTION

CONDITIONS -5M NaCl

5M NaCl, high salt

concentrations

are not appropriate!

Salt effect will lower solubility of Leachables. There will be no chemical interaction

between lipophilic polymer (surface) and 5M NaCl.

So, why this extraction?

EXAMPLE

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CUSTOMIZE REALISTIC EXTRACTION

CONDITIONS - POLYSORBATE

PS-80 generates high analytical background noise. It also suppress detectability of

Extractables.

PS-80 can be simulated by ethanol

or IPA/water mixtures!*

Tween could contain carry over

leachables that are sourced by

storage conditions and chemical

decomposition of PS-80.

*Dennis Jenke, Ph.D. A Means of Establishing and Justifying Binary Ethanol/Water Mixtures as Simulating Solvents in Extractables Studies,

Smithers/Rapra E&L USA 2015: Bethesda, MD; May 13, 2015.

EXAMPLE

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CUSTOMIZE REALISTIC EXTRACTION

CONDITIONS – POLYSORBATE RELATED

IMPURITIES

PS-80 is a polymeric mixture consisting from lipophilic monoesters condensed with

ethylene oxide.

EXAMPLE

Class Examples Source

Alcohols C4-8 PS-80 Breakdown

Aldehydes C6-10 PS-80 Breakdown

Ketones C5-C8 PS-80 Breakdown

Carbonic

acids

C4-8, C18, C16 PS-80 Breakdown

others Glycols-ethers PS-80 Breakdown

Leachables Antioxidant, fatty acids, fatty

alcohols,….

Interaction with

storage container

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RADIATION RESISTANCE

Validation of radiation regarding impact on extractable conc. level was

missed!

Dose: approx. 32 kGy

Extraction of Pt cured silicone tube at 50 °C for 24 h with WFI

Analyte Blank µg/mL µg/cm2 Compared to Blank

(LOQ) an increase

by factor of

Formaldehyde < LOQ1 21 7 233

Formiate < LOQ2 106 34 25

Acetate < LOQ2 18 14 9

1 0.03 µg/cm2 2 1.6 µg/cm2

EXAMPLE

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STANDARD EXTRACTION PROTOCOLS

- SELECTION OF EXTRACTION CONDITIONS

Misapplied use of plastic could cause chemical attack; it may

occur in several ways:

Disintegration or degradation of physical nature due to absorption,

permeation, solvent action, swelling…

Oxidation, where chemical bonds are attacked,

Hydrolysis, were e.g. ester bonds are attacked,

Radiation, were chemical reaction are triggered,

Thermal degradation,

Delamination, due to chemical attack.

In E&L studies those substances must be assessed that could

migrate into DP or process steam.

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STANDARD EXTRACTION PROTOCOLS

- RISK ASSOCIATED WITH SEP

Misconceptions increases the risk of faulty decisions.

Extraction conditions are too harsh:

Decomposition of the polymeric network, high levels of

oligomers are expected,

• Generation of high conc. levels of breakdown products.

Extraction conditions are too harsh:

• Screening methods do not discover all relevant

substances – combine with target analysis.

Material qualification can become not longer valid:

Under real use condition there is probably no risk on

leaching into the drug product.

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IMPROVEMENTS TO DISCUSS

– SOME CUSTOMIZATION NEEDED

Simulated migration studies with conditions that are selected

close to real process conditions would be the best practice to

asses risk from SUS such as bags, tubes, connectors and

filters ….

Extractables studies conditions should be selected to avoid

materials damage!

Extraction conditions selection must based on damage mode

studies and/or chemical resistance studies.

SGS is offering:

Chemical characterization (Extractables studies),

Simulated Use Studies (Leachables studies),

and Damage Mode Studies to support end-user materials

qualification process.

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LIFE INSPIRED

THANK YOU FOR YOUR ATTENTION

Visit us at our booth Hall 4.2 B30

Dr. Andreas NixdorfLife Sciences

Senior Scientist / Business Development Manager

SGS INSTITUT FRESENIUS GmbH

Extractables & Leachables – Impurity Profiling

H578

Kasteler Str. 45

D-65203 Wiesbaden

Phone: + 49 611 962 5903

E-mail : andreas.nixdorf@sgs.com

Web : www.sgs.com/lifescience