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Additional information

In vitro, the mutagenicity of ethylenediamine, +4PO was studied in an GLP and OECD guideline conform Ames test including both a standard plate test and a preincubation test (BASF, 40M0530/884153, 1988). The S. typhimurium strains TA98, TA100, TA1535 and TA1537 and the E.coli strain Wp2 uvrA were exposed to six concentration levels ranging from 0 -5000 μg/plate with and without metabolic activation by S9-mix. Negative and positive controls (2-aminoanthracene, N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitro-o-phenylendiamine, 9-aminoacridine) were included. The number of revertants did not increase when treated with the different test substance concentrations. According to the Ames test, ethylenediamine, +4PO is non-mutagenic.

To fullfil the requirements for genotoxicity of REACH Annex VIII-X, a grouping approach for NLP Polyols has been adapted. It is based on the report of Paul Illing Consultancy Services Ltd. and Marlin Consultancy (Illing and Barratt, 2007 & 2009) which identifies common breakdown products and functionalities within a given grouping category of NLP polyols. Accordingly, a read-across from ethylenediamine, +EO +PO (CAS# 26316 -40 -5) to ethylenediamine, +4PO (CAS# 102 -60 -3) is justified, as both polyols are grouped within the same NLP polyol category by Illing and Barrett.

For the GLP and OECD guideline conform chromosome aberration assay, V79 chinese hamster lung fibroblasts were exposed to five dose levels from 0 -2800 μg/ml with or without S9 -mix (ISOPA AT04950, 2008). Negative (vehicle) and positive controls (mitomycin c, cyclophosphamide) were included. Cytotoxicity was observed in the absence of S9 metabolic activation at 2100

μg/ml. None of the cultures treated with the polyol showed biologically relevant or statistically significant increased number of aberrant metaphases. Thus, under the experimental conditions chosen here ethylenediamine, +EO +PO is considered to be non-clastogenic in V79 cells.

For the GLP and OECD guideline conform HPRT, chinese hamster lung fibroblasts (V79) were exposed to seven dose levels (0 - 2800 μg/ml) of ethylenediamine, +EO +PO with and without S9 -mix (ISOPA T 0078744, 2009). Negative (vehicle) and positive controls (ethylmethansulfonate, dimethylbenzanthracene) were included. In the presence of metabolic activation, ethylenediamine, ethoxylated and propoxylated was not cytotoxic up to the highest dose tested which is equivalent to the limit dose of 10 mM. On the basis of the present study, the test substance does not cause an increase of the mutant frequencies in two independent experiments. According to the conditions chosen in the HPRT test, ethylenediamine, +EO +PO is non-mutagenic.

Overall, the available GLP and OECD guideline conform in vitro studies (Ames test, Chromosome Aberration Assay, HPRT) on genotoxicity are negative. Therefore, ethylenediamine, +4PO is considered to be non-mutagenic. According to REACH Annex VIII, an in vivo method was not applied to monitor genotoxicity, as all three conducted in vitro tests yielded negative results.

Short description of key information:

Genetic toxicity in vitro:

Ames test (GLP & OECD 421/422): not-mutagenic with and without S9-mix (S.typhimurium TA98, TA100; TA1535; TA1537 and  E.coli Wpa2 uvrA(BASF SE 40M0530/884153; 1988)

CA assay (GLP & OECD 473): non-clastogenic with and without S9-mix in V79 cells (ISOPA, AT04950, 2008)

HPRT (GLP & OECD 476): non-mutagenic with and without S9-mix in V79 cells (ISOPA T 0078744, 2009)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC and 1272/2008/EEC. In conclusion, classification of ethylenediamine, +4PO is not warranted.