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Summaries of the available fish, Daphnia and algal growth inhibition studies are presented below.

A 96 hour acute semi-static limit toxicity study in zebra fish (Brachydanio rerio) was performed to assess the toxicity of di-trimethylolpropane. The study was performed according to GLP and the OECD 203 Test Guideline. No mortalities were observed during the test, therefore the 96 hour LC50 was observed to be >1000 mg/L (the highest nominal concentration tested) (Olsson, 1992).

A static 48 hour definitive test was performed exposing Daphnia magna to a range of concentrations (1000 - 5000 mg/L) of di-trimethylolpropane. The study was performed in accordance with the OECD guideline 202 and according to GLP. The 48 hour EC50 was calculated to be 3560 mg/L (based on nominal concentrations) (Olsson, 2008)

A 72 hours algal growth inhibition limit study, test species Selenastrum capricornutum, was performed to determine the toxic effects of the test substance di-trimethylolpropane. The test was conducted according to OECD test guideline 201, and performed to GLP. The 72 hour EC50 value, based on growth rate, was observed to be >1000 mg/L (the highest nominal exposure concentration tested)

(Olsson, 1992).

In the chemical safety assessment performed according to Article 14(3) in connection with Annex I section 3 (Environmental Hazard Assessment) no hazard was identified. Therefore according to REACH Annex I (5.0) exposure estimation is not necessary for long-term toxicity to fish or aquatic invertebrates. Consequently, in accordance with Column 2 of REACH Annex IX, the studies do not need to be conducted as all identified uses of the substance are assessed as safe for the environment.

A summary of the available activated sludge study is presented below.

The toxicity of Di-Trimethylolpropane to activated sewage sludge was investigated in a respiration inhibition test according to OECD test guideline 209. Following three hours exposure to Di-Trimethylolpropane the EC50 was observed to be > 1000 mg/L (based on nominal exposure concentrations), the corresponding NOEC was observed to be 1000 mg/L (Knight, diTMP, 2010).