EXTRACTABLE LEACHABLES OVERVIEW, INDUSTRY REQUIREMENTS & AVECIA - - PowerPoint PPT Presentation

EXTRACTABLE LEACHABLES

OVERVIEW, INDUSTRY REQUIREMENTS & AVECIA APPROACH BY ARYO NIKOPOUR 12/1

• Definition
• Industry Guidance
• Extractables Method Development
• Extractables Method Validation
• Leachables Method Development
• Leachables Method Validation
• Extractables & Leachables Experience
• Case Studies

OUTLINE

Store and Protect The Drug Product WHAT DOES DRUG PACKAGING DO?

SUITABILITY OF CONTAINER CLOSURE SYSTEM

Protection

Suitability

for Use Safety Compatibility Performance

• Extractable: A chemical compound (volatile, semi-volatiles non-volatile and elemental

impurities) that gets extracted from a packaging component in a suitable solvent by utilizing

• ptimum extraction conditions (time and temperature).
• Extractable profile for a given packaging component, typically can be a chromatogram (GC-

MS, LC-MS and ICP-MS) representing all possible extractables.

• Extractable profile is established for all packaging components (resin, vial, foil-laminate) for

their consistent quality assurance. FDA DSaRMAC, May 2004

EXTRACTABLES

• Leachable is any chemical compound (volatile, non-volatile and inorganics ) that leaches into the

drug product formulation either from a packaging component or local environment on storage (time and temperature) through expiry of the drug product. An extractable can be a leachable.

LEACHABLES

Extractables

Leachables

Migrants

• The identity and concentration of recurring leachables in the drug product
• r placebo formulation should be determined through the end of the drug

products shelf life.

• In general, the levels of extractables should be greater than the levels of

leachables for the correlation to be considered valid.

• Evaluation of leachables in the drug product formulation in future routine

stability studies may not be needed when such a correlation exists.

Guidance for Industry: Container Closure System for Packaging Human Drugs and Biologics. May 1999

LEACHABLES

PHARMA/BIOPHARMA SUPPLY CHAIN

Raw Material Supplier Manufacturer Packagers & Distributor Retailer

Flow of Information and Traceability across

• Form a Team
• Role of Analytical Chemist

– Risk based approach DOE – Established study protocol – Execute experiments – Analyze data, determine AET – Identify E&L ≥AET – Submit report to team members for review

APPROACHES TO E&L STUDIES

Toxicologist Analytical Chemist Formulators Manufacturing RA/QA

REGULATORY BODIES AND ORGANIZATIONS

USP

Requirements: Guidelines:

Elastomeric Closures <381> Glass Containers <660> Plastic Containers <661> Biological Reactivity Tests – in vitro<87> Biological Reactivity Tests- in vivo <88> Extractables <1663> Leachables <1664>

Biocompatibility of Materials <1031>

MEDICAL DEVICE: ISO 10993 BIOCOMPATABILITY

• ISO 10993-1: Evaluation and testing in the risk management process
• ISO 10993-7: Ethylene oxide sterilization residuals
• ISO 10993-9: Framework for identification and quantification of potential

degradation products

• ISO 10993-12: Sample preparation and reference materials
• ISO 10993-13: Identification and quantification of degradation products from

polymeric medical devices

• ISO 10993-14: Identification and quantification of degradation products from

ceramics

• ISO 10993-15: Identification and quantification of degradation products from metals

and alloys

• ISO 10993-18: Chemical characterization of materials

MDI and DPI Drug Products:

Chemistry, manufacturing, and controls documentation. Suggested Testing: “The drug product should be evaluated for compounds that leach from elastomeric, plastic components or the coating of the container and closure system, such as…………. “…polynuclear aromatics , nitrosamines, monomers, plasticizers, accelerators, antioxidants and vulcanizing agents”

FDA GUIDANCE FOR INDUSTRY

• Determine the identification and concentration profile through the end of the drug product shelf life.
• Correlate with the extractables profile of the container and closure components (determined under

the various control extraction conditions).

FDA GUIDANCE FOR INDUSTRY

FDA GUIDANCE FOR INDUSTRY(USP)

Degree of Concern Associate with Route of Administration Likelihood of Packaging Component-Dosage Form Interaction High Medium Low Highest  Inhalation Aerosols  Injections and injectable Suspension  Sterile Powders and Powders for Injection  Inhalation Powder High  Ophthalmic Solutions  Transdermal Ointments and Patches  Nasal Aerosols and Sprays Low  Topical Solutions and Suspensions  Topical and Lingual Aerosols  Oral Solution and Suspensions  Topical Powders; Oral Powders  Oral Tablets  Oral Capsules

• Review DMF (Type III)
• Review CDER “Guidance for Container Closure Systems for Packaging of Human Drug and Biologics”
• Review Leachables and Extractables Testing Points to Consider form ITFG/IPAC-RS
• Review Guidance for MDI/DPI
• Leachables and Extractables Handbook, Wiley 2012
• OINDP/OPDP

STEP 1. LITERATURE SEARCH

• Is the container closure in contact with the formulation? Identify Material (i.e., HDPE, LDPE, etc.)
• Is the container closure in contact the patient’s mouth or mucosa?
• Secondary Container/Closure?
• Extractables/Leachables Potential?

STEP 2A. WHAT ARE THE CRITICAL COMPONENTS?

• Consult with a toxicologist for the Toxicity Evaluation of the major extractable.
• Determine the Permitted Daily Exposure (PDE).
• Finalize the target leachable compounds based on the quantitative data of extractable and

PDE data. Risk Assessments:

• Dose
• Duration
• Route of Administration
• Patient Population & Indication
• Special Population
• Women of Childbearing Potential

STEP 2B. TOXICITY EVALUATION

RISK BASED APPROACH TO E&L

• Safety

– Immunogenicity and toxicity

• Efficacy

– Leachable may cause lose of activity – Leachable may interact with product

• Quality

– Quality Attribute may change due to presence of leachables – May impact product stability

MDI DEVICE

Actuator

Contaminant

Canister

TYPICAL FOIL-LAMINATE COMPONENTS

A = Exterior layer Polyester/PP/PE (0.00048 inch) B = Inks C = Adhesive1

D = Aluminum Foil (0.00035 inch)

E = Adhesive2 F = Nylon/Polyester/ PP/PE(0.001 inch) G = Adhesive3 H = Interior layer Polyester/PP/PE (0.003 inch) DSaRMAC, May 05,2004

SINGLE USE SYSTEMS (SUS)

• SUS Components includes:

– Filters – Processing Containers (bags) – Tubing – Connectors – Gaskets – Valves All Product Contact surfaces have the potential to release extractables material into a process, In a biopharmaceutical process, a risk assessment for E&L should be make . Following Factors will affect the assessment: – Nature of Extractables – Process Fluid – Contact Time – Contact Temperature – Down Stream processes

EXTRACTABLES & LEACHABLES

USP Nomenclature PQRI-PODP Outcome Extractables Study USP<1663> Controlled Extraction Studies Identify Extractables as Tentative Leachables Simulation Study (Model Extraction Study) Simulation Study Extractables as probable Leachables (fewer extractables). Assessing Safety Risk from Contamination Leachables Study USP <1664> Migration Study Confirmed Leachables in support of Shelf Life Studies in Final CCS

Material Assessment Extractables Study AET Simulation Study AET Toxicological Assessment Leachables Study AET, SCT, TTC, QT

SOURCES OF EXTRACTABLES & LEACHABLES

Leachables

Manufacturing

SUS (Tubing, Filters , Bags)

Drug Substance Materials

Water, Excipients

Method(s) Container/ Closure Environment

Stability Migrants

• A controlled extraction study, is a qualitative and quantitative investigation of critical components of

CCS.

• Establish a basis for development and validation of analytical method in support of routine QC

testing.

• Establish a basis for development of Leachable studies.
• Allow for the “Correlation” of extractables and leachables.

Reference: Leachables and Extractables Handbook, Wiley 2012

STEP 3. ESTABLISH EXTRACTABLE PROFILE (CONTROL EXTRACTION STUDIES)

• 1. Extraction studies of critical component using solvents with different strengths:

 IPA (Soxhlet, Reflux)  Water (Pressurized Vessel)  Heptane (Soxhlet, Reflux) For semi permeable CCS, evaluate secondary CCS too.

What factors have impact on Diffusion:

 Time  Solvent  Temperature  Physical properties of the polymer

STEP 3. ESTABLISH EXTRACTABLE PROFILE (CONTROL EXTRACTION STUDIES)

STEP 3: CONTROLLED EXTRACTION STUDIES (CONT.)

Low Molecular Weight

• Monomers
• Solvent Residues
• Residues from polymer

treatment

• Polymer Breakdown

HS-GC/MS, HS-GC-FID

Screening, Target Analysis, Identification and Quantification

Mid Molecular Weight

• Lubricants
• Plasticizers
• Antioxidants
• Solvents

Direct Injection GC/MS GC-FID

Screening, Target Analysis, Identification and Quantification

High Molecular weight

• Plasticizers
• Antioxidants
• Anti-Slip Agents
• Fillers

LC/MS, LC-PDA Screening, Target Analysis, Identification and Quantification Elemental

• Metals

Alkali Earth Metals ICP/MS , ICP-OICES AAS, Screening, Target Analysis, Identification and Quantification

VOC SVOC NVOC Inorganics

STEP 3. ESTABLISH EXTRACTABLE PROFILE

The Proposed Specification for Extractables Lower detection limits required for nitrosamine, PAHs, and MBTs, they are consider as special case

Sources: ITFG/IPAC-RS “Leachables and Extractables Testing: Points to Consider”

Extractable Level in Component Tasks > 100 ppm Structure confirmed 20 –100 ppm Tentatively identified < 20 ppm Reported as unknown

• Safety Concern Threshold (SCT)

Low Risk Leachables Not Identified <0.15 µg/day

• Qualification Threshold (QT)

Assessment of Identified Leachables >5 µg/day

• Parenteral Drug Product:
• PQRI

The leachables as provided by the Chemistry , is evaluated based on individual leachables, concentration and estimated total daily dose, relative to the propose thresholds for safety evaluation

TOXICOLOGICAL THRESHOLD LEVEL

Class I Class II Class III Class IV Sensitizer Class IV Irritant Class V Genotoxicant Threshold Level (µg/day) 150 45 7.5 5 5 0.15 Proposal Class I No Genotox Class II No Genotox Class III Genotox M7 Threshold (µg/day) 50 if Systemic 5 If irritant/Sensitizer 1.5 To identify

ICH M7

• The ICH M7 Guideline was finalized in 2014 offering guidance on analysis of Structure Activity

Relationships (SAR) for genotoxicity. Furthermore, it is intended to resolve questions such as whether impurities with similar alerts that potentially have similar mechanism of action should not be combined in calculating a Threshold of Toxicological Concern (TTC) and whether the TTC may differ based on differences in the approved duration of use.

• To complement this ICH M7 Guideline an Addendum was finalized in 2017 to summarise known

mutagenic impurities commonly found or used in drug synthesis. The intent of this Addendum is to provide useful information regarding the acceptable limits of known mutagenic impurities/carcinogenic and supporting monographs.

Where,

AET: Analytical Evaluation threshold SCT: Safety Concern threshold Max #: Maximum number of doses a patient would take per day Component Wt. Weight of the component per dose (g/unit dose) UF: Uncertainty factor used to correct for source of uncertainty with the method

“Leachables and Extractables Handbook” Wiley publishing 2012

STEP 3: ANALYTICAL EVALUATION THRESHOLD (CONT.)

AET (µg/g)= [ (SCT(µg)/day)÷ (max# of doses/day)÷(wt. of component (g)/dose)] x (UF)

Component: Glass with Elastomeric Closure Delivery System: Injectable SCT: 1.5 µg/day

• No. of Doses: 1/day

Vial Volume: 1 mL Max # of Dose: 1 vial/day Uncertainty Factor: 50% AET= [1.5 µg/day] *[1day/1vial]* [1 vial/1 mL] * [0.5]= 0.75 µg/mL

EXAMPLE: ANALYTICAL EVALUATION THRESHOLD(CONT.)

Protocol Report Introduction about Container/Closure Introduction about Container/Closure Extraction Procedure Extraction Procedure Type of Solvents Type of Solvents Temperature/Duration Temperature/Duration Sample Size Sample Size Method of Analysis Method of Analysis Identify Surrogate Standards Results Internal Standards AET Evaluation Tentative ID of Peaks above AET level

STEP 3. CONTROL EXTRACTION DOCUMENTATION

• Based on the analytical and toxicological evaluation of the extractables from the control

extraction studies, the applicant should establish discriminatory test procedures and set appropriate acceptance criteria for the extractable profiles for routine testing for each critical component of the container closure system.

• This testing will provide continued assurance of the batch-to-batch consistency of the

composition and purity of the container and closure components. An extraction test should be performed on every incoming component batch, using water and other suitable solvents selected from the control extraction studies, to determine the individual and total extractables. *Guidance for industry: Nasal and Inhalation Solution

STEP 4. ROUTINE EXTRACTION

• Conduct Model extraction studies by choosing up to 3 different mediums representative of final

formulation (Placebo), e.g., buffer at various pH.

• Container closure will be filled with three formulation Placebo and stored at accelerated conditions,

e.g. 40ºC/75%, 50ºC.

• Samples are analyzed at initial, 1 week, 3 and 6 weeks.
• Representative methods from Controlled Extraction studies will be employed to analyze the samples

(LC/MS, GC/MS,..).

• Evaluate data and determine the potential Leachables.
• Model Extraction Studies will expedite the risk assessment for determination of leachable studies.

STEP 5. MODEL EXTRACTION(SIMULATION) STUDIES

Risk Assessments:

• Dose
• Duration
• Route of Administration
• Patient Population & Indication
• Special Population
• Women of Childbearing Potential

STEP 6. LEACHABLE STUDIES

• Apply extractable analytical methods to drug formulation and aged stability samples through shelf-

life.

• Optimize the methods to avoid interference from active and excipients.
• Do not apply Filtration
• In addition to Targeted Leachable method(s) apply Scan method in support of every sample
• Evaluate method characteristics during course of method development (e.g. Robustness and

Ruggedness)

• Qualify Reagents , at least two different manufacturers
• Identify suitable Internal Standards
• Use Surrogate Standards for Scanning Methods

STEP 6. LEACHABLE STUDIES (CONT.)

STEP 6. LEACHABLE STUDIES (CONT.)

Method Development Method Validation Specificity Specificity Linearity Linearity (QL- 300% Level of Concern) QL QL Accuracy At Level of Concern Accuracy At 50 to 200% of Level of Concern Precision Repeatability Precision Repeatability Intermediate Precision Solution Stability System Suitability System Suitability Robustness Robustness Establish Scan Method Add Scan Method parameters to Final Method

Follow ICH Q2(R1) for Quantitative Impurities

• Include Leachable Studies as part of Official Stability Studies in support of NDA/BLA and

ANDA/Biosimilar submission.

• Employ validated methodologies which could detect and accurately quantitate the potential

Leachables.

• Method(s) also need to monitor and screen for unexpected leachables (e.g. Migrants,…).

STEP 7. LEACHABLE STABILITY STUDIES (CONT.)

• Based on the chromatographic and spectroscopic data, a correlation between leachables and

extractables is established when each leachable can be linked qualitatively to a corresponding extractable, directly or indirectly.

STEP 8. ESTABLISH A CORRELATION BETWEEN E&L (CONT.)

• pMDI Valve
• Nasal Spray Actuator and Cap/Liner
• Injectable Bags
• PETE Bottles
• LDPE in Support of Inhalation Solution and Ophthalmic Products
• HDPE Bottles
• PVC/Foil Laminate in Support of Solid Dose Blister Pack
• Disposable Holding Bags (SUS)
• C/C for Parenteral Products (Small Molecules and mAb, oligonucleotides,..)
• PFS (mAb and small molecules)
• Medical Devices

NITTO AVECIA PHARMA SERVICES EXPERIENCE

CASE STUDIES

Controlled Extraction Studies for Rigid Tip Cap In Support of Glass Syringe Image of Syringe with Rigid Shield and Tip Cap

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Controlled Extraction Studies for Rigid Tip Cap In Support of Glass Syringe

• Controlled extraction studies are to systematically identify and quantify potential Extractables from

the Rigid Tip Cap.

• Controlled extraction studies were performed for the heat-induced extraction and solvent extraction

and analyzed by GC/MS, LC/MS and ICP/MS for volatile, semi-volatile, non-volatile and elemental impurities, respectively.

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Volatile Extractable Profile for Tip Cap Headspace-GC/MS Total Ion Chromatogram of Tip Cap A Sample

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Tip Cap Sample

Isobutylene Carbon Disulfide Thiirane Isopropanol Propene

AET = 0.94 µg/g

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

The identities of all extractables were verified against authentic reference standards except for 2-and 4-(1-Phenylethyl)-phenol, which were verified only by matching to NIST library.

AET Value In Support of Controlled Extraction Studies for Tip Cap

Proposed SCT (Safety Concern Threshold) = 1.5 µg/Day Max# of Dose = 1

• Wt. of Component = 0.8 g

UF (uncertainty factor) = 0.5

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

) / ( 94 . 5 . 8 . . 1 5 . 1 ) ( ) ( # max ) ( (µg/g) AET g µg UF dose g component

• f

wt day dose

• f

day µg SCT                                        Parameter Tip Cap AET (µg/g) 0.94 (µg/g)

Leachable Method Development & Validation

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Tungsten (W):

• Originates from the tungsten pins used to produce the channel through which the needle in the

syringe is mounted. Silicone Oil (Polydimethylsiloxane, PDMS):

• Widely used as a lubricant for the plunger in pre-filled syringes.
• Both silicone oil and tungsten have a tendency to interact with proteins leading to aggregation.
• The quality of a protein drug product is therefore affected by the presence of tungsten or silicone
• il from the syringe.

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Mechanism of silicone oil-induced protein aggregation: A protein can adsorb onto the hydrophobic silicone oil/water interface, which may or may not be reversible. Over time, protein can lose its conformation (denaturation), which is mostly irreversible. Denatured species can revert to the bulk and form aggregates with similar molecules in solution. (Source: Concise Encyclopedia of High Performance Silicones (2014), pp. 381-394)

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

• Non-GMP Study of Tungsten and Silicone Oil in Placebo-filled Syringe

Tungsten results are for the contents of each of nine syringes for each syringe type. Silicone oil results are for the contents of each of nine syringes for Syringe Type 2, and nine preparations with the combined contents of two syringes each for the other syringe types. QL is 2.0 µg/mL for placebo study.

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Summary:

• A method for determination of Silicone Oil in Protein Drug Product was developed with a

quantitation limit (QL) of 6.0 µg/mL.

• An optimized extraction procedure was required to obtain good recoveries of silicone oil from drug

product.

• Free Silicone Oil was found in various placebo-filled syringes at levels from less than QL to 73.2

µg/mL.

• Tungsten was found in the placebo-filled syringes from 0.10 to 2.75µg/mL.
• Other compounds found as extractables, were not detected as leachable in the Protein Drug Product.

CASE STUDY I:E&L STUDIES IN SUPPORT OF PFS

Introduction: – Single-Use BioProcess Bags are made of an advanced medical-grade film. – This film is a five-layer, 14 mil cast film and co-extruded polyolefin film. – The outer layer is a polyester elastomer coextruded with an EVOH barrier layer and an ultra-low density polyethylene product contact layer. – The layers of film also include: co-polyester ether (COPE), a blend of maleic anhydride modified and unmodified polyethylene, ethylene vinyl alcohol, and an ethylene alpha olefin material. – The commercial Bag sizes can very from less than 1 liter to greater than 1,500 liters. The volume of the Single-Use BioProcess Bags used for this study is 100 mL.

CASE STUDY II : LEACHABLE STUDIES IN SUPPORT OF SINGLE-USE BIOPROCESS BAG (SUS)

Techniques:

• 1. Ion Chromatography was used to quantitate levels of acids in bags
• 2. An Agilent GC/MS system was used to analyze the Headspace volatile organic compounds and the

Soxhlet solvent extractables for semi-volatile organics.

• 3. An API 4000TM LC/MS/MS system was used to analyze Soxhlet IPA/Heptane/H2O extractions for

polar, non-polar and aqueous extractables. Model Extraction Conditions:

• 1. All samples stored at 60ºC for 0, 1, 2, 3 and 4 weeks.
• 2. Model extraction solution were analyzed by IC, GC/MS and LC/MS.

CASE STUDY II: LEACHABLE STUDIES IN SUPPORT OF SUS

Results – Ion Chromatography

CASE STUDY II: LEACHABLE STUDIES IN SUPPORT OF SUS

Bag Boric Acid (µg/mL) Formic Acid (µg/mL) Citric Acid (µg/mL) Type I 17 28 Type II 56 7 52

• 21 Tentative Leachables found from Controlled extraction Study:

– Low molecular acids:

• Acetic acid, Boric acid, Formic acid, and Citric acid

– Three long chain aliphatic carboxylic acids

• Palmitic acid, Stearic acid, and Myristic acid

– Five low MW, non-antioxidant Aromatic compounds

• Dibutylphthalate, Dibutyl phthalate, 2, 4-Di-tert-butylphenol, 2, 5-Di-tert-butylphenol, 1, 3-Di-tert-butylbenzene, and

2, 2’-Bipyridyl

– Amides:

• Erucamide, Hexadecanamide, Octadecanamide, Oleamide

– Three Antioxidants:

• Irganox 1010, Irganox 1076, and Irganox 168

– Siloxane:

• Decamethylcyclopentasiloxane

CASE STUDY II LEACHABLE STUDIES IN SUPPORT OF SUS

Determination of the Structure of an Unknown Peak by MS/MS

CASE STUDY II: LEACHABLE STUDIES IN SUPPORT OF SUS

Structure of Unknown (Irgafos 168 degradant) Irgafos 168

Unknown Peak 1 2 4 5 6 7 8 2 1 7 8 9

251..20

3 LC/MS/MS ESI Negative Scan

RT= 27.97 min

The unknown peak is an Irgafos 168 Degradant Bis(2,4-di-tert-butylphenl) phosphate (bDtBPP)

Synthesis Characterization Method Development Method Validation Quantitation of the Sample in Bioprocess Bag

CASE STUDY II: LEACHABLE STUDIES IN SUPPORT OF SUS

P O OH O O

CONTROLLED EXTRACTION STUDIES IN SUPPORT OF Drug Product XYZ 13-MM LYOPHILIZATION STOPPERS AND Drug Product ZYX 20-MM SERUM STOPPERS Purpose: To Evaluate Extraction Profile of Elastomeric Closure Used in Support of the Lyophilized and Solution Product.

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13 MM LYOPHILIZATION AND 20 MM SERUM STOPPERS

Headspace Volatile Extractables Profile of Company XYZ 13-mm LYOPHILIZATION STOPPER

CASE STUDY III :CONTROLLED EXTRACTION STUDIES IN SUPPORT OF Z LYOPHILIZATION AND SERUM STOPPERS

XYZ 13-mm Lyo Stoppers Empty Vial (Blank) Heptane Cyclohexane Tetrahydrofuran Methylcyclopentane 3-Methyl-pentane 2-Methyl-pentane

80ºC for 30 min

Hexane

AET = 113 µg/g

Headspace Volatile Extractable Profile of ZYX 20-mm SERUM STOPPERS

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

Empty Vial (Blank)

80ºC for 30 min

Cyclohexane 3-Methyl-pentane Methylcyclopentane Hexane 2-Methyl-pentane ZYX 20-mm Serum Stoppers

AET = 42 µg/g

Headspace Volatile Extractable Results by GC/MS

CASE STUDY III : CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

No. Extractable Name XYZ 13-mm Lyo Stopper ZYX 20-mm Serum Stopper Extractable Conc. (µg/g) Extractable Conc. (µg/g) 1 2-Methyl-pentane 0.0426 0.0582 2 3-Methyl-pentane 0.114 0.176 3 Hexane 0.226 0.166 4 Methylcyclopentane 0.212 0.359 5 Tetrahydrofuran 0.282 6 Cyclohexane 0.102 0.109 7 Heptane 0.0426

Soxhlet Semi-volatile Extractable Profile

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

BHT Rubber Oligomer

Octadecane C18H38 Octacosane C28H58 Tetracosane C24H50 Docosane C22H46 Eicosane C20H42 XYZ 13-mm Lyo Stoppers & ZYX 20-mm Serum Stoppers Hexacosane C26H54

Soxhlet Semi-Volatile Extractable Results for XYZ 13-mm LYOPHILIZATION STOPPERS (Eight Semi-volatile Extractables found by GC/MS)

CASE STUDY III :CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

Semi-volatile Extractable Name Water Extraction Conc. (µg/g) Ethanol Extraction Conc. (µg/g) Heptane Extraction Conc. (µg/g) AET (µg/g) BHT 10.6 24.6 33.7 Rubber Oligomer 17.6 72.9 113 Octadecane 1.55 2.62 Eicosane 2.22 7.59 Docosane 3.62 17.5 Tetracosane 4.41 28.9 Hexacosane 3.64 28.6 Octacosane 5.05 55.8

Soxhlet Semi-volatile Extractable Results for ZYX 20-mm SERUM STOPPERS (Eight Semi-volatile Extractables found by GC/MS)

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

Semi-volatile Extractable Name Water Extraction Conc. (µg/g) Ethanol Extraction Conc. (µg/g) Heptane Extraction Conc. (µg/g) AET (µg/g) BHT 16.9 179 12.6 Rubber Oligomer 52.2 304 42.0 Octadecane 1.94 7.24 Eicosane 3.47 15.8 Docosane 6.26 29.7 Tetracosane 7.80 45.4 Hexacosane 7.35 45.9 Octacosane 9.11 84.8

Non-volatile Extractable Profile By LCMS/MS

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

Elemental Extractable Profile by ICP-MS Elemental Extractable Name Heptane Extraction Conc. [XYZ 13-mm Lyo Stopper] (µg/g) Heptane Extraction Conc. [ZYX 20-mm Serum Stopper] (µg/g) Mg 0.045 0.050 Ca 0.15 0.10 Sr 0.0011 0.0011 Cu 0.00040 0.00010 Zn 0.0030 0.0015 Ni 0.0040 0.0030 Ta 0.00000050 0.00000050 V 0.00015 Not Detected AET (µg/g) 113 42

• Conclusions:

– 7 Volatile Extractables found but All are less than AET. – 8 Semi-volatile Extractables found but only BHT and Rubber oligomers are greater than AET. – No non-volatile Extractable found. – 8 Elemental Extractables found but all are less than AET.

CASE STUDY III: CONTROLLED EXTRACTION STUDIES IN SUPPORT OF 13-MM LYOPHILIZATION STOPPERS AND 20-MM SERUM STOPPERS

• “Heat-induced Extractable Profile Study In PVC films Using Headspace GC/MS Methodology”;
• Mai Zhou, Kareem Chehade, Aryo Nikopour; 234th American Chemical Society National Meeting, Poster ID: 1109066, Division of

Analytical Chemistry, 19 August 2007, Boston, MA,

• “Residual Solvents Screening By Utilization Headspace - GC/MS”;
• Aryo Nikopour, Mai Zhou, Ruggero Picci, Darren Lorenz; 2007 AAPS Annual Meeting and Exposition, Poster ID: 3451, Nov. 11-15, 2007,

San Diego, CA, USA

• “Heat-Induced and Soxhlet Solvent Extraction Profile Study In Foil Laminate Film Using GC/MS Methodology”; Mai Zhou, Kareem A. H.

Chehade, Aryo A. Nikopour; 2008 AAPS Annual Meeting and Exposition, Poster ID: 3225, Nov. 16-20, 2008, Atlanta, Georgia, USA

• Testing of Biopharmaceuticals for USP Class 1, 2 and 3 Residual Solvents Using GC/MS, Mai Zhou, John Cui, Aryo Nikopour, Paul

Maffuid, BIOZONA 2009, Arizona’s Annual Bioscience Conference, April 7, 2009, Phoenix, Arizona

• “Heat-Induced and Soxhlet Solvent Extraction Profile Study in Medical Rubber Component used for Container Closure System Using

GC/MS Methodology”, Mai Zhou, John Cui, Joseph Bordas-Nagy, Aryo A. Nikopour, 2009 AAPS Annual Meeting and Exposition, Poster ID: T2106, Nov. 10, 2009, Los Angeles, California, USA

• “Heat-Induced and Soxhlet Solvent Extraction Profile Study in HDPE Bottle Used for Container Closure System Using GC/MS

Methodology”; Mai Zhou, Dong Zhao, John Cui, Joseph Bordas-Nagy, Aryo A. Nikopour; 2010 AAPS Annual Meeting and Exposition, Poster ID: W5005; Nov. 17, 2010, New Orleans, Louisiana, USA

• “Determination of Melamine, Ammeline, Ammelide and Cyanuric Acid in Pharmaceutical Raw Materials by LC-MS/MS”; John Cui, Mai

Zhou, Aryo Nikopour; 2013 AAPS Annual Meeting and Exposition, Poster ID: AM-13-1266; Nov. 10, 2013, San Antonio, Texas, USA

• “Heat-Induced and Soxhlet Solvent Extraction Profile Study in Elastomeric Closure Using GC/MS Methodology”; Mai Zhou, Spencer

Huang, Robert Lee, Aryo Nikopour; 2013 AAPS Annual Meeting and Exposition, Poster ID: AM-13-0934; Nov. 10, 2013, San Antonio, Texas, USA

EXRACTABLES/LEACHABLES POSTERS

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