Reaction products of fatty acids, tall oil and fatty acids, C18 unsaturated, trimers and fatty acids, C18 unsaturated, dimers with (9Z)-octadec-9-en-1-amine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

No study exists for the Reaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine. Read across was performed to another fatty acid / primary alkyl amine salt: fatty acids, C18 -unsatd., dimers, compds. with coco alkylamines (CAS 68647 -95 -0). Besides the structural similarities, both substances are liquids with a high molecular weight above 1000g/mol, are insoluble in water, highly lipophilic, have a negligible vapour pressure of 0.0001Pa, and are skin sensitizers and skin irritants. For the HPRT test, read across was performed to Oleylamine (CAS 112 -90 -3, CMA 1985), which comprises 49% of the test substance and is presumably the hazard inducing component. A detailed justification for the performed read across is attached in IUCLID chapter 13.

Gene mutation in bacteria

In a gene mutation assay in bacteria (Ames) according to OECD471 and GLP (BASF 2012) fatty acids, C18-unsatd., dimers, compds. with coco alkylamines did not lead to an increase in the number of his + or trp+ revertants in both, the standard plate test and in the preincubation test either without S-9 mix or after the addition of phenobarbital / β-naphthoflavone. induced rat liver S9. Concentrations up to 5000µg/plate (standard assay) or up to 1000µg/plate (pre-incubation assay) were tested in Salmonella strains TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli WPA2 uvrA.

In vitro Micronucleus test in mammalian cells

Fatty acids, C18-unsatd., dimers, compds. with coco alkylamines in THF did not induce micronuclei in V79 cells of the Chinese hamster in vitro in the absence and presence of phenobarbital / ß-naphtoflavon induced rat liver S9. Four independent experiments were performed. In Experiment IA the exposure period was 4 hours with metabolic activation. In Experiment IB and IC the exposure period was 4 hours without S9 mix. In Experiment II the exposure period was 4 hours with and 24 hours without metabolic activation. The cells were prepared 24 hours after start of treatment with the test item. In each experimental group two parallel cultures were set up and at least 1000 cells per culture were scored for micronuclei. The following concentrations were used, but only concentrations showing no excessive cytotoxicity as determined via the proliferation index were used for scoring. Cytotoxicity was observed at app. 40µg/ml with S9, at app. 16µg/mL without S9 after 4h, and above 5µg/mL without S9 after 24h.

4 hrs exposure without S9 (concentrations from 2 independent experiments):

1st: 1.3, 2.5, 5.0, 7.5, 10.0, 12.5, 15.0, 20.0, 30.0, 40.0µg/mL

2nd: 0.5, 1.0, 1.3, 1.5, 2.0, 2.5, 5.0, 7.5, 10.0, 12.5, 15.0, 30.0µg/mL

24hrs exposure without S9:

0.04, 0.08, 0.16, 0.31, 0.63, 1.3, 2.5, 5.0, 10.0, 20.0µg/mL

4hrs exposure with S9 (2 independent experiments):

1st: 0.4, 1.0, 2.6, 6.6, 16.4, 41.0, 102.4, 256.0, 640.0, 1600.0, 4000.0µg/mL

2nd: 2.5, 5.0, 10.0, 20.0, 30.0, 40.0, 50.0, 60.0, 80.0, 100.0µg/mL

Visible precipitation of the test item in the culture medium was observed in Experiment IA in the presence of S9 mix at 41.0 µg/mL and above at the end of treatment. Phase separation was observed in Experiment IB in the absence of S9 mix at 30.0 µg/mL

and above and in the presence of S9 mix in Experiment II at 40.0 µg/mL and above. No relevant influence on osmolarity or pH value was observed.

In Experiment the first experiment in the absence of S9 mix statistically significant increases in micronucleated cells (3.6, 3.05 and 2.23 %) above the historical control data range (0.15 - 1.50 %) were observed in a concentration range of 2.5 - 7.5 µg/mL. In the confirmatory statistically significant increases (0.65 - 1.28 % micronucleated cells) clearly within the range of the historical control data

were observed after treatment with 1.5, 2.0, 5.0, 7.5 and 10.0µg/mL. Thus the positive finding of the first experiment could not be confirmed. After 24h in the absence of S9 mix no relevant increase in micronucleated cells was observed. In the presence of S9 mix two statistically significant increases in micronucleated cells were observed after treatment with 40.0 and 60.0 µg/mL (0.65 and 0.60 %) in the second experiment, which were clearly in the range of the historical control data of 0.05 - 1.70 % and thus considered biologically irrelevant. Statistical significance was due to a very low frequency in the concurrent control of only 0.15%.

Gene mutation in mammalian cells

The test substance, oleylamine, was tested in the CHO/HGPRT mutation assay in the absence and presence of Aroclor-induced rat liver S-9 mix. The first assay was conducted at dose levels of 2.0, 1.5, 1.0, 0.5, and 0.1nl/ml (without S9), and at 9.0, 8.0, 7.0, 6.0,

and 5.0 nl/ml (with S-9). The repeat experiment consisted of two independent parallel experiments, conducted at 2.5, 2.25, 2.0, 1.5 and 1.0 nl/ml (without S-9) and at 10, 9.5, 9.0, 8.0, and 7.0 nl/ml (with S-9). The cells were incubated for 5h with the test substance, followed by a 7 -9 day expression period and a 7day selection period using 6 -thioguanine as the selection agent.

The positive and negative controls were within the historical control range. In the absence and presence of metabolic activation, oleylamine induced a more than 2 -fold increas in mutant colonies relative to solvent control cultures in the highes dose of the first experiment. No dose-response was oberved, and the result could not be confirmed in two additional independent parallel experiments. Thus the increased mutant frequency at a single dose observed in the first assay only, is considered an anomaly and as not relevant. Therefore, the test article is considered negative in the CHO/HGPRT mutation assay in the presence and absence of metabolic activation.

Additional evidence

Several mutagenicity studies exist for related alkylamines. No signs of mutagenicity were found. Study summaries and references can be found in the EU risk assessment report for primary alkyl amines (2008).

Coco alkylamine (CAS 61788 -46 -3), Tallow alkylamine (CAS 61790 -33 -8), Octadecylamine (CAS 124 -30 -1), and Oleylamine (CAS 112 -90 -3) did not cause gene mutations in bacteria (Ames test).

Oleylamine also produced negative results in the HPRT assay mentioned above, and in a mouse lymphoma assay.

Oleylamine also did not cause chromosomal aberrations in CHO cells in vitro.

In vivo tests for chromosome aberrations (mouse micronucleus tests) were performed with Oleylamine and Tallow alkylamine. Though clinical signs and toxicity up to lethality were observed with both substances, no increase in aberrant cells was reported.

Short description of key information:
Ames (read across from 68647-95-0): negative (BASF 2012, GLP, OECD 471)
Ames (read across from 112-90-3): negative (EU RAR 2008)
MNT in vitro (read across from 68647-95-0): negative (BASF 2012, GLP, OECD487)
CA (read across from 112-90-3): negative (EU RAR 2008)
HPRT (read across from 112-90-3): negative (CMA 1985, GLP, equivalent to OECD 467)
MLA (read across from 112-90-3): negative (EU RAR 2008
MNT in vivo (read across from 112-90-3): negative (EU RAR 2008)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Fatty acids, C18 -unsatd., dimers, compds. with coco alkylamines did not induce gene mutations in bacteria or chromosome aberrations in mammalian cells. Oleylamine, did not cause gene mutation in bacteria, mammalian cells, nor chromosome aberrations in vitro or in vivo. Several further negative mutagenicity studies exist for related primary alkyl amines.

Thus, there is no indication, that the Reaction products of Fatty acids, tall-oil, compds. with oleylamine and Fatty acids, C18-unsatd., trimers, compds. with oleylamine cause genetic damage. Therefore the substance has not to be classified according to EU criteria (67/548/EEC) or CLP/EU-GHS for this hazard.