The Importance of E&L Testing: Regulatory Perspectives

Introduction:

In the highly regulated pharmaceutical, biopharmaceutical, and medical device industries, ensuring patient safety and product integrity is paramount. Extractables and leachables (E&L) testing has emerged as a critical component of regulatory compliance. These tests identify and quantify potentially harmful chemical substances that may migrate from packaging, manufacturing systems, or delivery devices into the product, potentially impacting patient health or product efficacy. This article explores the significance of E&L testing, its regulatory landscape, methodologies, and its pivotal role in safeguarding public health.

Extractables and leachables testing has become an indispensable component of pharmaceutical and medical device development, driven by stringent regulatory requirements and patient safety imperatives. In an era where regulatory agencies demand comprehensive safety data before product approval, extractables and leachables testing serves as the cornerstone of quality assurance for any material that comes into contact with pharmaceutical formulations or biological fluids.

The pharmaceutical industry has witnessed significant regulatory evolution over the past two decades, with agencies worldwide recognizing that chemicals migrating from packaging materials, delivery systems, or device components can compromise product integrity and patient safety. This reality has elevated extractables and leachables testing from a peripheral concern to a central pillar of product development strategy.

From infusion sets and prefilled syringes to inhalation devices and transdermal patches, every drug-contact material must undergo rigorous evaluation to identify potentially harmful compounds that might migrate into the final product. Regulatory bodies including the FDA, EMA, and Health Canada have established clear expectations that manufacturers must meet through systematic extractables and leachables testing programs.

To learn more about E&L regulatory frameworks, visit:

• https://resolvemass.ca/el-testing-in-the-united-states/
• https://resolvemass.ca/el-regulatory-requirements-for-medical-devices/
• https://resolvemass.ca/extractables-vs-leachables-in-medical-devices/
• https://resolvemass.ca/ich-q3e-guideline-for-extractables-and-leachables/

Learn the basics of E&L testing in simple terms – Extractables and Leachables (E&L) Testing

Summary

Extractables and leachables testing is a critical quality assurance process that ensures pharmaceutical products, medical devices, and drug delivery systems remain safe throughout their lifecycle. This comprehensive guide explores:

• Regulatory requirements from FDA, EMA, and ICH for extractables and leachables testing
• Why E&L testing is mandatory for drug-contact materials and medical device components
• Key differences between extractables (what can migrate) and leachables (what does migrate)
• Industry best practices for conducting comprehensive extractables and leachables testing
• Risk assessment frameworks that guide testing protocols and acceptance criteria
• Analytical techniques including GC-MS, LC-MS, and ICP-MS used in E&L studies
• Real-world compliance challenges and how laboratories address regulatory expectations

Extractables are chemical compounds that can be extracted from a material under aggressive laboratory conditions, such as exposure to aggressive solvents or high temperatures. They represent the worst-case scenario of chemical migration.

Extractables represent the universe of compounds that could potentially migrate from a material. These studies use:

• Aggressive solvents (polar, non-polar, and intermediate polarity)
• Elevated temperatures
• Extended contact times
• Multiple extraction techniques

The goal of extractables testing is to create a comprehensive chemical profile that identifies worst-case migration scenarios.

Leachables are compounds that migrate into a drug product or patient-facing system under normal use conditions. These substances often originate from:

• Primary Packaging Materials (e.g., glass vials, plastic containers, rubber stoppers).
• Manufacturing Components (e.g., tubing, filters, and processing equipment).
• Delivery Devices (e.g., inhalers, syringes, and infusion pumps).
• Use real-time and accelerated stability conditions
• Monitor migration over the product shelf life
• Provide data directly relevant to patient exposure
• Validate that extractables findings translate to real-world scenarios

Aspect	Extractables	Leachables
Conditions	Aggressive, exaggerated	Realistic, actual use
Medium	Multiple solvents	Actual drug product
Purpose	Identify potential compounds	Quantify actual migration
Timing	Early development	Throughout lifecycle
Temperature	Elevated	Storage/use conditions

The Need for E&L Testing

E&L testing is crucial for several reasons:

• Patient Safety: Leachables can pose toxicological risks, including carcinogenic, mutagenic, and teratogenic effects.

See how E&L testing protects patients – The Role of E&L Testing in Ensuring Patient Safety 

• Regulatory Compliance: Stringent guidelines from the FDA and other regulatory bodies mandate comprehensive E&L evaluations.
• Product Quality Assurance: Prevents drug-product interactions, ensuring stability and efficacy.
• Global Market Access: Complying with U.S. and international regulations facilitates seamless product approval and distribution.

Curious about how much E&L testing costs? Read this – Cost of Extractables and Leachables Testing

2: Regulatory Perspectives in the United States

Regulatory agencies, including the FDA and USP (United States Pharmacopeia), emphasize rigorous E&L testing to mitigate risks associated with chemical migration. Key guidelines include:

FDA Guidance

1. Container Closure Systems: FDA guidance requires robust assessments of container closure systems to ensure they do not adversely interact with drug products.
2. Quality by Design (QbD): Encourages integrating E&L considerations into product development.

The U.S. Food and Drug Administration has issued several guidance documents addressing extractables and leachables testing:

Container Closure Systems

The FDA expects comprehensive evaluation of container closure systems for injectable drugs, requiring:

• Complete characterization of extractables profiles
• Leachables monitoring throughout stability studies
• Safety qualification of detected compounds
• Control strategies for identified risks

Orally Inhaled and Nasal Drug Products (OINDP)

The FDA’s OINDP guidance specifically addresses extractables and leachables testing for inhalation products, where patient exposure via the respiratory route creates unique safety concerns. Requirements include:

• Systematic extractables studies using multiple solvents
• Controlled extraction studies mimicking product use
• Leachables assessment in routine batches
• Toxicological risk assessments for all detected compounds

EMA Requirements

The European Medicines Agency has established clear expectations for extractables and leachables testing through various guidelines:

• Guideline on plastic immediate packaging materials: Addresses polymeric materials in direct contact with medicinal products
• Quality and equivalence guidance: Requires E&L data for generic drug applications
• Nitrosamine guidance: Expanded E&L considerations to include nitrosamine formation potential

ICH Guidelines

International Council for Harmonisation guidelines provide globally harmonized approaches:

• ICH Q3D on Elemental Impurities: Requires assessment of metal leachables from manufacturing equipment and components
• ICH M7 on Mutagenic Impurities: Establishes frameworks applicable to leachable compound risk assessment

ISO Standards

ISO 10993 series for medical devices includes specific requirements for extractables and leachables testing:

• ISO 10993-12: Sample preparation and reference materials
• ISO 10993-18: Chemical characterization of medical device materials

United States Pharmacopeia (USP)

• USP <1663>: Focuses on assessing extractables from pharmaceutical packaging and delivery systems.
• USP <1664>: Addresses the evaluation of leachables and their impact on drug product quality.

ICH Guidelines

• ICH Q3D: Emphasizes the control of elemental impurities in drug products.
• ICH Q8: Highlights the importance of understanding material compatibility in pharmaceutical development.
• E&L requirements for medical devices: https://resolvemass.ca/el-regulatory-requirements-for-medical-devices/

Other Regulatory Bodies

Organizations such as the International Organization for Standardization (ISO) and ASTM International also provide standards for E&L testing methodologies.

3: Why Extractables and Leachables Testing Is Mandatory

Extractables and leachables testing is mandatory because chemical migration from materials can cause adverse patient reactions, compromise drug efficacy, and violate regulatory safety standards. The consequences of inadequate testing extend beyond regulatory non-compliance.

Patient Safety Imperatives

Historical incidents have demonstrated the critical importance of extractables and leachables testing:

1. Toxicity events: Leached plasticizers have caused adverse reactions in patients receiving IV medications
2. Efficacy impacts: Active pharmaceutical ingredients can be absorbed by packaging materials, reducing drug potency
3. Sensitization reactions: Certain leached compounds trigger allergic responses in sensitive populations
4. Carcinogenic concerns: Some packaging components release compounds with carcinogenic potential

Regulatory Compliance

Regulatory submissions require comprehensive extractables and leachables testing data:

• Pre-market approval: New Drug Applications (NDAs) and Abbreviated New Drug Applications (ANDAs) must include E&L data
• Post-market surveillance: Changes to packaging or manufacturing require updated E&L assessments
• International harmonization: Global marketing requires meeting multiple regulatory standards simultaneously

Quality Assurance

Extractables and leachables testing forms part of the overall quality system:

• Ensures consistency across manufacturing batches
• Validates supplier material quality
• Supports change control processes
• Provides data for risk management activities

4: Applications of E&L Testing

E&L testing is integral to:

• Pharmaceutical Packaging: Ensures compatibility and safety of packaging materials.
• Biopharmaceutical Manufacturing: Validates that equipment does not release harmful compounds.
• Medical Devices: Evaluates device safety by identifying potential leachables. See how we test leachables in medical devices – Case Study: Identifying and Mitigating Leachables in Medical Devices 
• Combination Products: Assesses interactions between drug formulations and delivery devices.

Learn how E&L testing applies to respiratory devices – Extractables and Leachables (E&L) Testing for Inhalation and Nasal Drug Products: A Step-by-Step Guide

5: Key Components of an Extractables and Leachables Testing Program

A comprehensive extractables and leachables testing program includes material characterization, systematic extraction studies, leachables monitoring, analytical method development, and toxicological risk assessment. Each component builds upon the others to create a complete safety evaluation.

Risk Assessment

Every extractables and leachables testing program begins with risk assessment:

• Material composition analysis: Understanding polymer types, additives, and processing aids
• Contact conditions evaluation: Assessing temperature, duration, and contact surface area
• Route of administration consideration: Different exposure routes require different safety thresholds
• Patient population factors: Pediatric, geriatric, and immunocompromised populations may have heightened sensitivities

Extractables Studies

Systematic extractables testing establishes the baseline:

Solvent Selection

• Polar solvents: Water, dilute acids/bases
• Non-polar solvents: Hexane, isopropanol
• Intermediate polarity: Ethanol, acetone
• Simulating solvents: Matching drug formulation polarity

Extraction Conditions

• Temperature ranges: Room temperature to 60°C or higher
• Duration: 24 hours to several weeks
• Reflux extraction for aggressive assessment
• Controlled extraction mimicking actual use

Leachables Studies

Leachables monitoring provides real-world validation:

• Accelerated stability studies: 40°C/75% RH for up to 6 months
• Real-time stability studies: Labeled storage conditions for shelf life
• Routine testing protocols: Ongoing monitoring across commercial batches
• Multiple time points: Initial, intermediate, and end-of-shelf-life testing

Analytical Techniques for Extractables and Leachables Testing

Technique	Application	Compounds Detected
GC-MS	Volatile and semi-volatile organics	Plasticizers, antioxidants, monomers
LC-MS	Non-volatile organics	Polymer fragments, stabilizers
ICP-MS	Elemental impurities	Metals from catalysts, equipment
Headspace GC	Volatile organics	Residual solvents, low MW compounds
IC	Ionic species	Anions, cations
NMR	Structural elucidation	Unknown compound identification

Safety Qualification

All detected compounds above analytical thresholds require toxicological evaluation:

• Database searches: Checking against toxicology databases (ECHA, EPA, etc.)
• Structure-activity relationship (SAR) analysis: Predicting toxicity from chemical structure
• Threshold of Toxicological Concern (TTC): Applying safety thresholds for low-level exposures
• Comparative assessments: Evaluating against established safety data

6: E&L Testing Methodologies

Effetive Extractables and leachables Testing involves a systematic approach comprising:

1. Study Design

• Material Selection: Identify components that may release extractables or leachables.
• Simulated Conditions: Mimic worst-case and real-use scenarios.

How Much Does Extractables and Leachables (E&L) Testing Cost in 2025? Click here to make your testing plan cost-efficient.

2. Analytical Techniques

• Gas Chromatography-Mass Spectrometry (GC-MS): Ideal for volatile and semi-volatile compounds.
• Liquid Chromatography-Mass Spectrometry (LC-MS): Suitable for non-volatile and polar compounds.
• Inductively Coupled Plasma Mass Spectrometry (ICP-MS): Detects elemental impurities.
• Fourier Transform Infrared Spectroscopy (FTIR): Identifies organic functional groups.

3. Toxicological Risk Assessment

• Identifies and quantifies risks posed by detected leachables.
• Determines permissible exposure levels (PELs) based on toxicological data.

4. Data Interpretation and Reporting

• Comprehensive documentation of test results.
• Correlation of findings with regulatory requirements.

7: Challenges in E&L Testing

• Complexity of Materials: Diverse polymeric materials complicate extractable profiling.
• Analytical Sensitivity: Detecting trace levels of leachables requires cutting-edge instrumentation.
• Data Interpretation: Distinguishing between harmful leachables and benign substances demands expert analysis.

Still have questions about E&L testing? Check this –  Top Questions About E&L Testing Answered 

8: Emerging Trends in E&L Testing

Current trends in extractables and leachables testing include focus on nitrosamines, increased scrutiny of prefilled syringes and combination products, advanced analytical techniques, and sustainability considerations in material selection. The regulatory landscape continues evolving in response to scientific advances and safety concerns.

Nitrosamine Contamination

Recent nitrosamine findings have transformed extractables and leachables testing approaches:

• Root cause investigations: Identifying nitrosamine formation pathways in materials
• Preventive strategies: Selecting materials less prone to nitrosamine formation
• Enhanced testing: Adding nitrosamine-specific analytical methods
• Risk mitigation: Implementing control strategies across supply chains

Advanced Drug Delivery Systems

Complex delivery systems create unique challenges:

• Combination products: Medical devices with drug components require comprehensive E&L programs addressing both device and drug regulations
• Wearable devices: Extended wear times increase leachables exposure
• Smart delivery systems: Electronic components introduce new material types requiring evaluation

Sustainability and Green Chemistry

Industry is balancing safety with environmental responsibility:

• Alternative materials: Bio-based polymers requiring new E&L approaches
• Reduced additive formulations: Minimizing extractables sources while maintaining material performance
• Recyclable systems: Evaluating safety of materials designed for sustainability

1. Advanced Analytical Technologies

• High-resolution mass spectrometry and multi-dimensional chromatography enhance detection capabilities.

2. Risk-Based Approaches

• Emphasizing risk assessments over exhaustive testing to optimize resources.

3. Digitalization

• Integration of AI and machine learning for predictive toxicology and material behavior modeling.

4. Sustainability

• Developing eco-friendly packaging materials to reduce extractable risks.

See how we solved a real E&L challenge – Case Study: Enhancing Product Safety Through Comprehensive E&L Testing 

9: Why Choose ResolveMass Laboratories for E&L Testing?

ResolveMass Laboratories Inc. stands out as a leader in extractables and leachables (E&L) testing in the United States due to:

1. State-of-the-Art Facilities

• Equipped with advanced analytical instruments, including GC-MS, LC-MS, and ICP-MS.

2. Expert Team

• Our team comprises experienced chemists, toxicologists, and regulatory specialists.

3. Comprehensive Services

• From study design to toxicological assessments, we provide end-to-end solutions.

4. Regulatory Expertise

• In-depth understanding of FDA, USP, and ICH guidelines ensures compliance.

5. Tailored Solutions

• Custom testing protocols designed to meet client-specific requirements.
• Need a custom study design? We’ve got you – Customized E&L Testing Solutions for Your Industry

See the full list of E&L testing services offered – Extractables and Leachables services

10: Case Studies: Real-World Applications of Extractables and Leachables Testing

Real-world extractables and leachables testing case studies demonstrate how comprehensive evaluation programs prevent safety issues, support regulatory approvals, and maintain product quality throughout the lifecycle. These examples illustrate practical application of regulatory requirements.

Injectable Product Container Closure System

A pharmaceutical manufacturer developing a new injectable formulation encountered challenges:

• Initial findings: Extractables study revealed multiple compounds from rubber stoppers
• Risk assessment: Toxicological evaluation identified one compound exceeding safety thresholds
• Material change: Switched to alternative stopper formulation with different cure system
• Re-testing: Confirmatory extractables and leachables testing showed acceptable profile
• Outcome: Successful regulatory approval with documented material change rationale

Metered Dose Inhaler Development

An OINDP product required comprehensive E&L evaluation:

• Complex materials: Multiple plastics, elastomers, and metals in device
• Simulating use: Controlled extraction mimicking patient actuation patterns
• Leachables monitoring: Throughout stability in actual formulation
• Metals focus: Particular attention to valve components per ICH Q3D
• Result: Complete characterization supporting global registrations

Prefilled Syringe Safety Assessment

A biosimilar manufacturer needed E&L data for prefilled syringe presentation:

• Protein interaction concerns: Assessing potential for leachables to interact with biologic
• Silicone oil evaluation: Characterizing lubricant layer composition and migration
• Tungsten assessment: Evaluating metal exposure from manufacturing process
• Comparability: Demonstrating E&L profile comparable to reference product
• Approval: Data supported demonstration of biosimilarity

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11: Future Directions in Extractables and Leachables Testing

The future of extractables and leachables testing will involve increased automation, predictive modeling, harmonized global standards, and integration with overall pharmaceutical quality frameworks. These developments promise to make E&L evaluation more efficient while maintaining rigorous safety standards.

Technological Advances

• High-resolution mass spectrometry: Improved unknown identification capabilities
• Automated data processing: AI/ML tools for compound identification and library matching
• Non-targeted screening: Comprehensive detection without predefined target lists
• Miniaturized systems: Reduced sample requirements for expensive biologics

Regulatory Evolution

Expected regulatory developments include:

• Additional guidance documents: More specific requirements for emerging product types
• International harmonization: Greater alignment between regional agencies
• Risk-based flexibility: More tailored approaches based on product-specific factors
• Real-world evidence: Incorporating post-market surveillance data

Industry Standardization

Collaborative initiatives aim to:

• Shared databases: Industry-wide toxicology and safety information repositories
• Standard methods: Harmonized analytical approaches reducing redundant development
• Material master files: Pre-approved materials simplifying product development
• Best practice sharing: Industry associations facilitating knowledge transfer

ResolveMass Laboratories Inc.: Comprehensive Scientific Expertise You Can RelyResolveMass Laboratories Inc. is a trusted Canadian contract research organization offering a wide spectrum of specialized services spanning polymer synthesis, advanced analytical testing, and custom organic synthesis. With over a decade of experience supporting pharmaceutical, biotech, and industrial clients, we bring scientific precision and regulatory insight to every project. Our core capabilities include Polymer Synthesis and Characterization, Peptide Characterization, Organic Synthesis, Nitrosamine Testing and Analysis, PFAS Testing, and Extractable & Leachable Studies, as well as a broad suite of analytical techniques such as HPLC, GC-MS, MALDI-TOF, NMR, and FTIR. 

Our multidisciplinary team includes chemists, analytical scientists, and regulatory experts with advanced academic and industry backgrounds. We excel at developing customized, high-quality solutions—whether you need innovative polymer designs, impurity profiling, or confirmatory testing that meets global regulatory standards. 

Clients across North America choose ResolveMass Laboratories for our deep technical knowledge, commitment to quality, and ability to deliver reproducible, reliable data that drives confident decision-making. When precision, innovation, and trust matter—ResolveMass is your partner of choice. 

Need full-service support? Check this out – Choosing the Right E&L Testing Service Provider 

Conclusion: The Critical Importance of Extractables and Leachables Testing

Extractables and leachables testing stands as an essential element of pharmaceutical quality assurance and regulatory compliance, protecting patients from potentially harmful chemical migration while enabling safe product innovation. As regulatory expectations continue evolving and analytical capabilities advance, companies must maintain comprehensive extractables and leachables testing programs that meet both current requirements and anticipate future developments.

The regulatory landscape demands that pharmaceutical manufacturers, medical device companies, and their suppliers invest in robust extractables and leachables testing strategies from early development through post-market lifecycle management. Success requires not only technical analytical capabilities but also deep regulatory understanding, toxicological expertise, and strategic planning to integrate E&L considerations throughout product development.

For organizations navigating the complex requirements of extractables and leachables testing, partnering with specialized laboratories provides access to the expertise, instrumentation, and regulatory knowledge necessary for successful product development and approval. The investment in comprehensive extractables and leachables testing ultimately protects both patients and business interests by ensuring products meet the highest safety standards while avoiding costly delays or post-approval complications.

FAQs on E&L Testing: Regulatory Perspectives

Reference

1. Thomas F. Understanding Evolving E&L Regulations. In the Lab eNewsletter. 2020 Jan 8;15(1).
2. Vas G, Fleck L, Comstock K, Cole J. Extractable and leachable testing for pharmaceutical packaging, finished pharmaceutical products, and medical devices: an analytical perspective.
3. Ramamoorthy S, Chong NS, Hotha KK. Strengthening Extractable & Leachable Study Submissions: Best Practices to Avoid Regulatory Deficiencies. American Journal of Analytical Chemistry. 2024 Dec 5;15(12):368-94.