Advancing non-animal testing approaches for a toxic-free environment

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Publication date: 14 August 2025

Source: Green Deal Projects Support Office

The transition to non-animal testing methods—known as New Approach Methodologies (NAMs)—is gaining momentum as Europe pushes toward a toxic-free environment. Supported by the European Green Deal and aligned with the Zero Pollution Action Plan, NAMs offer faster, more ethical, and more human-relevant alternatives to traditional animal testing. These include in vitro testing, in silico modelling, and organ-on-a-chip technologies.

NAMs are essential to modernising chemical safety assessments and meeting the EU’s sustainability and public health goals. However, regulatory and technical challenges remain. To overcome these, the EU is developing a roadmap to phase out animal testing and has launched several Green Deal-funded projects—such as ALTERNATIVE, PANORAMIX, LifeSaver, SCENARIOS, ZeroPM, and PROMISCES—that are advancing science, building policy support, and demonstrating how NAMs can address complex risks like chemical mixtures and PFAS.

This article highlights key projects, policy developments, and the necessary steps for regulatory reform. It outlines how collaboration between researchers, regulators, and industry will be critical to fully integrating NAMs into EU legislation and achieving a more ethical, efficient, and sustainable approach to chemical safety.

Introduction

Creating a toxic-free environment has become a priority in regulatory toxicology, the science of assessing the safety of chemicals. Traditionally, this has involved testing on animals, but there has been a growing push to find better, more ethical alternatives, called New Approach Methodologies (NAMs). These include:

In vitro testing: experiments performed outside a living organism and typically in a controlled laboratory environment with glassware such as test tubes and flasks
In silico modelling: computer-based simulations and artificial intelligence (AI) models that predict how toxic a chemical might be; and
Organ-on-a-chip: small devices that mimic how human organs work, enabling researchers to see how chemicals affect the body.

These methods are not only more humane, but they also give faster and more accurate results than animal testing. These efforts are aligned with the EU’s broader sustainability goals, particularly the Zero Pollution Action Plan, which aims to reduce harmful chemical exposure through innovative, non-animal testing methods. In particular, the EU’s commitment to the 3Rs principle—Replacement, Reduction, and Refinement of animal use—underpins its support for NAMs as a cornerstone of ethical science. In addition, the European Union is currently developing a ”Roadmap Towards Phasing Out Animal Testing for Chemical Safety Assessment”, the implementation and progress of which can be supported by the development of these new methods.

Regulatory toxicology’s shift away from animal testing is being driven by scientific progress, ethical concerns, and new laws. In Europe, initiatives and policies like the European Green Deal and the Zero Pollution Action Plan are leading the way, aiming to protect both people and the planet, while also encouraging innovation in how we test chemical safety. The EU recognises that rules need to change to support non-animal testing, so that safety checks keep up with today’s scientific progress.

The European Green Deal has supported multiple projects to advance and promote these methods – read on for a few notable examples:

Understanding the risks of chemical mixtures

PANORAMIX aims to create new scientific tools and practical methods for assessing the health and environmental risks caused by the real-life mixtures of chemicals found in our water, food, and the human body.

The project is working on a new way to test how chemicals affect the human body without using animals. It uses an innovative approach, combining in vitro testing with advanced chemical analytical tools, to identify the chemicals that are the major drivers of the adverse effects on human health. PANORAMIX uses in vitro tests that are linked to in vivo adverse outcomes in humans, to assess toxicity of complex chemical mixtures, as well as for studying if and how chemicals act together in mixtures. These studies help researchers to figure out if a chemical mixture is harmful or not, in a faster, more predictable and more ethical way than traditional animal testing.

Their initial tests on complex chemical mixtures, extracted from the environment, food, and human blood, showed that the known chemicals in these mixtures behaved as expected: their effects added up based on their concentrations in lab tests related to health. However, when researchers compared the total biological impact of the full mixture to what could be explained by just the known and measured chemicals, they found a big gap. Less than 1% of the observed effect could be explained by the chemicals they had identified and quantified.

This means that the vast majority of the toxic effect is likely caused by chemicals that are either not detected at all or are present in such low amounts that they fall below detection limits - but they still contribute to the overall biological impact. This highlights the hidden risk of chemical mixtures: even when individual chemicals seem harmless or undetectable, their combined presence can still have significant health effects.

Testing drugs for maternal and foetal safety

The LifeSaver project is changing how we test new drugs by promoting innovations in drug testing that do not involve animal cruelty. Traditional drug safety evaluations often involve extensive animal testing, raising ethical concerns and sometimes failing to predict human-specific effects. The LifeSaver initiative aims to replace these outdated methods with NAMs, thereby ensuring that drug safety assessments minimise animal suffering whilst being more relevant to human biology, instead of animals.

Assessing the safety of ‘forever chemicals’

Lastly, several Green Deal-funded projects use NAMs in their study of per- and polyfluoroalkyl substances (PFAS). Commonly referred to as “forever chemicals”, these substances are used in everyday items like non-stick pans, waterproof clothing, food packaging, and firefighting foam because they resist heat, water, and oil.

However, these same properties make PFAS harmful to the environment and potentially dangerous to human health. They don’t break down easily and can build up in people, animals, and nature over time.
Traditionally, testing the safety of these chemicals has relied on animal testing, which is slow, expensive, and raises ethical concerns. That’s why projects like SCENARIOS, ZeroPM, and PROMISCES are using New Approach Methodologies (NAMs) instead.

These include:

High-throughput screening (fast, automated lab testing),
Computer modelling, and
Advanced bioassays (cutting-edge lab tests using living cells or tissues).

These modern tools help scientists better understand how PFAS affect health and the environment without using animals and support smarter, faster decisions by regulators.

Policy and regulation recommendations from the projects

The European Commission is committed to phasing out animal testing in regulatory toxicology, aligning with the objectives of the Green Deal, REACH (Registration, Evaluation, Authorisation, and Restriction of Chemicals), and the ASPIS Cluster. These frameworks emphasise the need for innovative and scientifically validated alternatives to animal testing.

Within the context of the Green Deal, several updates are worth noting. Firstly, the Zero Pollution of Persistent and Mobile Substances (ZeroPM) project has developed a Regulatory Watch, which provides policy updates and developments, as well as advocacy relating to topics such as chemical safety, advocating for the regulatory acceptance of NAMs in assessing PFAS risks.

Secondly, in October 2024, the Green Deal Health Cluster, which includes ALTERNATIVE, LifeSaver, PANORAMIX, PROMISCES, SCENARIOS and ZeroPM projects, published a policy brief, called “Achieving zero pollution by 2050 needs regulatory change: a call for policy support of New Approach Methodologies (NAMs)”. This brief emphasises the need for next-generation regulation of PFAS using emerging exposure, chemical grouping, non-standard data, and non-animal methodologies:

Emerging exposure: refers to the identification and assessment of new or unexpected exposures to hazardous substances.
Chemical grouping: involves categorising chemicals based on their structural similarities and shared properties.
Non-standard data: refers to data that is not typically collected or available in standard datasets, and therefore may require additional extraction and processing to be used in research or regulatory contexts
Non-animal methodologies: alternative testing methods that do not involve the use of animals.

The policy brief also highlights that the current dependence of chemical regulations on animal testing is a major challenge impeding the achievement of the Green Deal’s zero-pollution ambition. It lists several policy recommendations, including support for using exposure, non-standard data and chemical grouping for regulatory action, support for improving and validating NAMs, support leveraging uncertainty assessment for NAM recognition, support for the provision of aid for the evolution of a NAM-based next-generation regulation, and support for the initiative to develop a European roadmap to an animal-free regulatory system.

Challenges in integrating NAMs into regulatory frameworks

Despite scientific advancements, the adoption of NAMs in regulatory toxicology faces several challenges:

Regulatory acceptance: Many regulatory bodies require extensive validation before approving new testing methodologies.
Standardisation issues: NAMs must be standardised to ensure consistency and reliability across testing environments.
Data gaps: Some NAMs still require further development to comprehensively assess complex toxicological endpoints.
Resistance to change: The regulatory and scientific communities must undergo a cultural and modal shift to fully embrace non-animal methods.

Overall, addressing regulatory barriers is key to accelerating the adoption of NAMs.

The role of the Green Deal-funded projects in shaping policy

Projects funded under the European Green Deal are helping to drive this transition. These initiatives are shaping the conversation around regulatory reform, by generating robust scientific evidence and engaging with policymakers. Their work is essential in building confidence in NAMs, advocating for updated guidelines, and paving the way for a more ethical, efficient, and human-relevant approach to chemical safety.

Key NAMs European activities

Collaboration between universities, businesses, and regulators is key to accelerating the adoption of NAMs. Joint efforts have helped share knowledge through events such as:

ASPIS Open Symposium: Provides a platform for integrating the results of the three EU Horizon projects (ONTOX, PrecisionTOX, and RiskHunt3r) to ensure the application of NAMs in regulatory toxicology.
Green Deal Science-Policy Dialogue: Engages policymakers, scientists, and industry leaders to promote NAM adoption.
European Commission Roadmap towards phasing out animal testing for chemical safety assessments: The Commission plans to publish this roadmap in the first quarter of 2026 at the latest. Phasing out animal testing will require substantial time – for instance, there are currently no available approaches that are sufficient for developing and validating methods. Consequently, the roadmap foresees an implementation phase for the years to come.

Partnerships between governments and private companies (thanks to EPAA) are key to driving innovation in NAMs. By combining their resources, knowledge, and funding, these collaborations help turn scientific research into real-world solutions that regulators can use.

The roadmap for complete regulatory acceptance of NAMs

Achieving a fully animal-free regulatory toxicology framework requires a strategic, multi-phase roadmap. This roadmap must integrate scientific, regulatory, infrastructural, and educational components to ensure the widespread adoption and acceptance of New Approach Methodologies (NAMs). The European Commission’s “Roadmap Towards Phasing Out Animal Testing for Chemical Safety Assessments” provides a foundational structure for this transition,

Scientific validation: NAMs must undergo rigorous validation to demonstrate reliability, reproducibility, and relevance. The development of internationally accepted validation protocols is essential. The Commission supports initiatives like revising OECD Guidance Document 34 and standardising emerging technologies such as organ-on-chip.
Harmonisation of regulations: Global alignment is critical. The roadmap emphasises harmonised regulatory standards across jurisdictions, with the OECD playing a central role in facilitating consensus and mutual recognition of NAMs.
Investment in infrastructure: More investment is needed to ensure labs have the right tools and technology to use NAMs, and an economic ecosystem to provide the services. The roadmap also calls for building an economic ecosystem that supports NAM services and innovation.
Training and capacity building: The Commission highlights the importance of education and training for scientists, regulators, and stakeholders. This includes capacity building through partnerships such as EPAA, PARC, and ASPIS, and the involvement of EU-NETVAL

Key milestones on this roadmap include the development of internationally accepted validation protocols, harmonised regulatory standards, and increased investment in training and infrastructure. More specifically, international cooperation is essential for the widespread adoption of NAMs. The EU, the US, and other global stakeholders should collaborate to share best practices, harmonise regulations, and promote scientific advancements in NAMs. The Commission’s roadmap encourages consultation activities and working groups to foster cross-border dialogue and coordination.

Milestones and implementation

The roadmap outlines short- to long-term milestones, including:

The establishment of working groups focused on human health, environmental safety, and change management.
The development of consultation strategies and stakeholder engagement mechanisms.
The publication of the full roadmap by Q1 2026, with implementation phases extending beyond that date.

This strategic approach aims to accelerate the transition to an animal-free regulatory system while maintaining high safety and scientific integrity standards.

Conclusion

The transition to non-animal testing approaches is a crucial step toward achieving a toxic-free environment. NAMs are not only more ethical but also give results that are more relevant to humans and are often faster and more efficient. Green Deal-funded projects have played a pivotal role in advancing NAM research and advocating for regulatory changes, and will continue to be highly relevant for future developments.

Even though there are still challenges, continued funding, updated regulations, and better public understanding will help NAMs become more widely used. By working together among researchers, policymakers, and industry stakeholders, we can accelerate the shift towards a fully animal-free regulatory toxicology framework.

As science and regulations evolve, non-animal methods will play a bigger role in keeping people and the environment safe, while supporting the EU’s goals for ethical and sustainable science. Consequently, the EU’s leadership reinforces its global role in promoting humane, science-based regulatory practices that align with its environmental and public health objectives.

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