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All endpoints in this chapter were covered by read-across of studies of 1,4-bis[(2-ethyl-6-methylphenyl)amino]anthraquinone. This read-across is justified as both substances have very similar chemical structures, the main physicochemical parameters (vapour pressure, water solubility and dissociation constant) are in the same range and further all available toxicological studies for both substances revealed the same results. Therefore a read-across for other physicochemical, ecotoxicological or toxicological endpoints seems to be justified. More details about the read-across justification are displayed in a detailed document attached to the Annex of the CSR.

The studies were performed to assess the acute toxicity to aquatic organisms under static and semi-static conditions. The aquatic organisms were exposed to a limit test concentration dissolved in dilution water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and different filter system. All results are expressed in terms of nominal concentrations. The values were below the quantification limit of the HPLC analysis.

Acute toxicity to fish (Danio rerio): No toxic effects against fish were observed after 96h exposure at a limit test concentration of 100 mg/L.

Toxicity to aquatic invertebrates (Daphnia magna): After 48 hours of exposure, an EC50 > 100 mg/l was reported. No toxic effect was observed.

Toxicity to aquatic algae and cyanobacteria: After 72 h no toxic effects against algae were observed at a limit test concentration of 100 mg/l.

Toxicity to microorganisms: The test item showed 0 % respiration inhibition of activated sludge after a test period of 3 hours at the highest concentration of 1000 mg/L. Therefore a 3 hrs-EC50 higher than 1000 mg/L was calculated for the test substance.

All toxicity studies mentioned above are classified as acceptable and satisfy the guideline requirements for the toxicity studies on acute toxicity to aquatic organisms.