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If released in water,12-ketochenodeoxycholic acid is expected to biodegrade quite quickly, according to results of the ready biodegradability test. Thus, the test item, although it is not used in a detergent, can be considered a surfactant as defined in Regulation 648/2004/EC: “any organic substance and/or mixture used in detergent, which has surface-active properties and which consist of one or more hydrophilic and one or more hydrophobic groups of such a nature and size that is capable of reducing the surface tension of water, and of forming spreading or adsorption monolayers at the water-air interface, and of forming emulsions and /or microemulsions and/or micelles, and of adsorption at water-solid interfaces”. Therefore, a surfactant shall be considered as biodegradable if the level of biodegradability is at least 60% within 28 days, irrespectively from 10-day window principle. In this test, carried out according to OECD TG 301F, ursodeoxycholic acid reached 53.4% of degradation after the 10-d window, and 82% after 28 days. Therefore 12-ketochenodeoxycholic acid, basing on a read-across approach, is considered readily biodegradable.