Amines, C12-14-tert-alkyl, C14-18 and C18-unsatd.-alkyl phosphates

Endpoint summary

Additional information

General considerations

Based on the intrinsic properties of reaction products of alcohols, C14-18, C18 unsat., esterified with phosphorus pentoxide and salted with amines, C12-14, -tert-alkyl, Water Accommodated Fractions (WAFs) were prepared for testing. This procedure is in accordance with the OECD Guidance document on aquatic toxicity testing of difficult substances and mixtures (Series on testing and assessment - Number 23; OECD, 2000). The term WAF is applied to aqueous media containing only the fraction of multi-component substances that is dissolved and / or present as a stable dispersion or emulsion. These fractions are prepared individually and not by serial dilution of a single stock WAF. As the term indicates, only a fraction of the total mass of multi-component substances responsible for the composition may be present in the WAF. The "loading rate" is the mass to volume ratio of the mixture to medium used in the preparation. The obtained LL50/ EL50 values are comparable to LC50/ EC50 values, as well as the NOELR (No Observable Effect Loading Rate) to NOEL (No Observable Effect Concentration).

Fish

Oncorhynchus mykiss was used as test organisms in an experiment according to OECD Guideline 203 (Shacklady, 2001). Based on the outcome of the range finding study (no mortality and no sub-lethal effects up to 1000 mg/L), the definitive test was conducted as limit test with a single concentration of 1000 mg/L in two replicates. A static test regime was applied and a control group was included.

After 96 h, the NOEL was determined as 1000 mg/L WAF, together with a LL50 of > 1000 mg/L WAF. The toxicity data available demonstrate that the substance is not toxic to fish even at the highest test substance concentration tested. Based on these results and due to animal welfare reasons, no chronic investigation with fish is intended, which is in accordance to REACH, Annex IX, Section 9.1.6 (column 2).

Aquatic invertebrates

Vryenhoef and Mullee (2012) used Daphnia magna as freshwater representative in an experiment according to OECD Guideline 202.

Based on the results, the definitive test was conducted with the following test concentrations: 10, 18, 32, 56 and 100 mg/L (WAFs, each with four replicates).

The EL50(48h) value for the test substance was determined as 91 mg/L WAF with 95 % confidence limits of 76 - 110 mg/L WAF. The NOELR(48h) amounts to 56 mg/L WAF. 24h-values are reported as: EL50 > 100 mg/L WAF and the NOELR was 100 mg/L WAF.

Aquatic algae

The toxicity towards the unicellular algae Pseudokirchneriella subcapitata was investigated according to OECD Guideline 201 (Vryenhoef and Mullee, 2012). Based on the results of two range-finding experiments, the definitive test was conducted with loading rates of 0.10, 0.32, 1.0, 3.2 and 10 mg/L WAF. Three replicates were prepared for each treatment group.

The following results were revealed: ErL10(72h): 1.9 mg/L WAF, ErL20(72h): 3.5 mg/L WAF, ErL50(72h): 10 mg/L WAF based on growth rate inhibition. The corresponding No Observed Effect Loading Rate (NOELR) was 1.0 mg/L WAF, the Lowest Observed Effect Loading Rate (LOELR) was 3.2 mg/L WAF. Based on yield inhibitions (biomass), the EbL10(72h) amounts to 0.86 mg/L WAF, EbL20(72h): 1.3 mg/L WAF and EbL50(72h): 2.4 mg/L WAF with 95 % CL of 1.9 - 2.9 mg/L WAF. Aquatic microorganisms

The toxicity towards aquatic microorganisms was investigated according to the conditions of OECD Guideline 209 (Clarke, 2012). Activated sludge, collected at an aeration tank of a predominantly domestic sewage treatment plant (STP) was used as inoculum. Synthetic sewage was used as respiration substrate. Based on the findings in the range-finding test and review, the definitive test was conducted with test concentrations of 100, 320 and 1000 mg/L to obtain a NOEC. 3,5-dichlorophenol was used as reference, resulting in an EC50(3h) value of 11 mg/L. No statistically significantly toxic effects were shown at the test concentrations of 100 and 320 mg/L. Effects were observed at 1000 mg/L. Hence the NOEC amounts to 320 mg/L after 3 h exposure.