REACH test methods updated: new OECD Test Guidelines 321 (bioconcentration in Hyalella azteca), 252 and 253 (endocrine disruption), in vitro methods for eyes and skin sensitisation, dustiness of nanoforms under EN 17199
Commission Regulation (EU) 2025/2573 of 18 December 2025, published on 19 December, amends the Annex to Regulation (EC) No 440/2008: it updates the in vitro methods for serious eye damage and skin sensitisation, adds three ecotoxicological methods and dustiness standards for nanoforms. According to the recitals, the aim is also to reduce the number of animals used in testing (Directive 2010/63/EU).
- New in the fate and ecotoxicity table: OECD TG 321 — Hyalella azteca bioconcentration test (HYBIT), OECD TG 252 — rapid estrogen activity in vivo assay (REACTIV) and OECD TG 253 — short-term juvenile hormone activity screening assay with Daphnia magna (JHASA), all from 2024. HYBIT has an uptake and a depuration phase; the rate constants k1 and k2 and the bioconcentration factors BCFSS and BCFK are derived.
- The in vitro section for serious eye damage now covers TG 437, 438, 460, 491, 492, 492B, 494 and 496 plus the defined approaches of TG 467, and the one for skin sensitisation TG 442C, 442D and 442E in their 2024 versions plus the defined approaches of TG 497 (2023). TG 403 (acute inhalation toxicity), 442B (LLNA: BrdU-ELISA or FCM) and 493 (hrER estrogen receptor) were also updated.
- Dustiness of nanoforms of substances: the table now lists EN 17199-1 to -5:2019 (requirements and choice of methods, rotating drum, continuous drop, small rotating drum, vortex shaker) alongside EN 15051-1:2013, EN 15051-2:2016 and EN 15051-3:2013.
- The full descriptions of methods A.13 (pyrophoric properties of solids and liquids), B.2 (acute inhalation toxicity) and B.70 (estrogen receptor) were deleted — part 0 now refers to the equivalent international methods instead. The regulation entered into force on the twentieth day after publication.