This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union .

This Regulation shall be binding in its entirety and directly applicable in all Member States.

Done at Brussels, 18 December 2025.

For the Commission

The President

Ursula VON DER LEYEN

( 1 ) OJ L 396, 30.12.2006, p. 1 , ELI: https://data.europa.eu/eli/reg/2006/1907/oj .

( 2 ) Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) ( OJ L 142, 31.5.2008, p. 1 , ELI: https://data.europa.eu/eli/reg/2008/440/oj ).

( 3 ) Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes ( OJ L 276, 20.10.2010, p. 33 , ELI: http://data.europa.eu/eli/dir/2010/63/oj ).

( 4 ) OECD Test Guideline 442D: In Vitro Skin Sensitisation: Assays addressing the Adverse Outcome Pathway Key Event on Keratinocyte activation (2024) https://doi.org/10.1787/9789264229822-en ; OECD Test Guideline 467: Defined Approaches for Serious Eye Damage and Eye Irritation (2024) https://doi.org/10.1787/28fe2841-en ; OECD Test Guideline 496: In vitro Macromolecular Test Method for Identifying Chemicals Inducing Serious Eye Damage and Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage (2024) https://doi.org/10.1787/970e5cd9-en .

( 5 ) OECD Test Guideline 252: Rapid Estrogen Activity In Vivo (REACTIV) assay (2024) https://doi.org/10.1787/54066090-en ; OECD Test Guideline 253: Short-term Juvenile Hormone Activity Screening Assay using Daphnia magna (JHASA) (2024) https://doi.org/10.1787/03cb5c08-en ; OECD Test Guideline 321: Hyallela azteca Bioconcentration Test (HYBIT) (2024) https://doi.org/10.1787/8ac30c4e-en .

( 6 ) OECD Test Guideline 403: Acute Inhalation Toxicity (2024) https://doi.org/10.1787/9789264070608-en .

( 7 ) OECD Test Guideline 442B: Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA or –FCM (2024) https://doi.org/10.1787/9789264090996-en .

( 8 ) OECD Test Guideline 442C: In Chemico Skin Sensitisation: Assays addressing the Adverse Outcome Pathway key event on covalent binding to proteins (2024) https://doi.org/10.1787/9789264229709-en .

( 9 ) OECD Test Guideline 442E: In Vitro Skin Sensitisation: In Vitro Skin Sensitisation assays addressing the Key Event on activation of dendritic cells on the Adverse Outcome Pathway for Skin Sensitisation (2024) https://doi.org/10.1787/9789264264359-en .

( 10 ) OECD Test Guideline 492: Reconstructed human Cornea-like Epithelium (RhCE) test method for identifying chemicals not requiring classification and labelling for eye irritation or serious eye damage (2024) https://doi.org/10.1787/9789264242548-en .

( 11 ) OECD Test Guideline 492B: Reconstructed Human Cornea-like Epithelium (RHCE) Test Method for Eye Hazard Identification (2024) https://doi.org/10.1787/0d603916-en .

( 12 ) OECD Test Guideline 493: Performance-Based Test Guideline for Human Recombinant Estrogen Receptor (hrER) In Vitro Assays to Detect Chemicals with ER Binding Affinity (2024) https://doi.org/10.1787/9789264242623-en .

COMMISSION REGULATION (EU) 2025/2573 of 18 December 2025 amending Regulation (EC) No 440/2008 as regards the test methods, to adapt them to technical progress ANNEX