Hexylamine

Endpoint summary

Additional information

In vitro tests:

Ames

In a GLP conform bacterial reverse mutation assay, performed according to OECD guideline 471/472 (1983/Revised Draft March 1996), the test substance (analytical purity 99.0 area %) was investigated in the Ames test using the plate incorporation test and the pre-incubation test. Thereby, Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and the Escherichia coli strain WP2 uvrA were utilised. The assay was performed with and without metabolic activation (S9 mix of Aroclor 1254 -induced rat livers). The test substance was dissolved in DMSO. Each concentration, including the controls (positive and negative), was tested in triplicate. The test item was tested at the following concentrations: Standard plate test: 20; 100; 500; 2,500 and 5,000 µg/plate with and without metabolic activation; Preincubation test: 125; 250; 500; 1,000 and 2,000 µg/plate with and without metabolic activation. No test substance precipitation was found. An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test with and without metabolic activation and consistently a negative result was obtained. Cytotoxic effects (reduced his-negative or trp-negative background growth, decrease in the number of his-positive or trp-postive revertants, reduction in the titer) was observed in the standard plate test from about 2500 µg/plate onward. In the preincubation assay cytotoxicity was found from about 500 - 1,000 μg/plate onward. The positive control substances (2-aminoanthracene for all strains with metabolic activation; N-methyl-N’-nitro-N-nitrosoguanidine for the strains TA 100 and TA 1535, 4-nitro-o-phenylendiamine for the strain TA 98; 9-aminoacridine hydrochloride hydrate for the strain TA 1537 without metabolic activation; all substances dissolved in DMSO) produced valid responses.

HPRT

The read across substance n-Butylamine was tested in a HPRT test according to OECD Guideline 476. Therefore, mouse lymphoma L5178Y cells were used and treated with the test substance with and without metabolic activation (S9 mix from Aroclor 1254 induced rat livers). Two experiments were conducted using the following test substance concentrations: Experiment 1: 0, 50, 100, 210, 280, 320, 360, 400, and 500 µg/mL (evaluated); Experiment 2: 0, 50, 200, 300, 400, 425, and 450 µg/mL (evaluated). 4-nitroquinoline-N-oxide (without S9) and benzo(a)pyrene (with S9) were used as positive controls. Cells were exposed to the test substance for 3 hours at a temperature of 37 +/- 1 °C followed by a 7 days expression period. After this period the cells were incubated with 6-thioguanine for 12 days for selection of resistant cells. As a result, n-Butylamine did not induce mutation at the hprt locus of L5178Y mouse lymphoma cells under test conditions employed in this study. This included treatments up to highly toxic concentrations in the two independent experiments.

The same test was performed on another read across substance (Octylamine). Test concentrations were: Experiment 1: 0, 10, 20, 30, 40, 50, 60, 70, 75, 80, 90, 100 µg/mL (evaluated); Experiment 2: 0, 10, 20, 30, 40, 50, 55, 60, 65, 70, 72.5, 75, 80, 90 µg/mL (evaluated). As a result, N-octylamine did not induce mutation at the hprt locus of L5178Y mouse lymphoma cells either under the test conditions used. Again, the test substance was tested up to highly toxic concentrations.

In vivo tests:

Micronucleus

For the detection of chromosome aberrations induced by the read across substances butylamine and octylamine hydrochloride, two Micronucleus assays were conducted according to OECD Guideline 474 and in compliance with GLP.

The micronucleus test on butylamine was conducted on ICR mice. Doses of 125, 250, and 500 mg/kg bw were used, applied by oral gavage as concentrations of 6.25, 12.5, and 25 mg/mL butylamine in distilled water at dose volumes of 20 mL/kg bw. Erythrocytes were evaluated 24, 48 and 72 hours after treatment. No induction of micronuclei was observed. It was concluded that butylamine was not clastogenic under the test conditions chosen.

A second read across study was conducted on octylamine hydrochloride. For this micronucleus assay, male NMRI mice were used. 75, 150 and 300 mg/kg bw of octylamine in water were applied by gavage to 5 mice per dose. Cyclophosphamide (CPP) and Vincristine Sulphate (VCR) were applied intraperitoneal as positive controls. Samples of the bone marrow were taken 24 hours and 48 hours after the treatment. In general 2000 polychromatic erythrocytes (PCEs) from each of the male animals of every test group were evaluated and investigated for micronuclei. The normochromatic erythrocytes (NCEs) which occurred were also scored. The number of normochromatic erythrocytes containing micronuclei did not differ to any appreciable extent in the negative control or in the various dose groups at any of the sacrifice intervals. No inhibition of erythropoiesis induced by the treatment of mice with Octylamine hydrochloride was detected. The ratio of polychromatic to normochromatic erythrocytes was always in the same range as that of the control values in all dose groups. Thus, under the experimental conditions chosen here, the test substance Octylamine hydrochloride has no chromosome-damaging (clastogenic) effect nor does it lead to any impairment of chromosome distribution in the course of mitosis (aneugenic activity) in bone marrow cells in vivo.

Justification for selection of genetic toxicity endpoint
Several studies on different types of genetic toxicity were used for assessment.

Short description of key information:
Ames:
The test substance was not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay.
HPRT:
Two read across substances (butylamine and octylamine) were not mutagenic in the HPRT test.
Micronucleues Test:
Two read across substances (butylamine and octylamine hydrochloride) did not induce chromosome aberrations in the Micronucleus assay.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for genetic toxicity under Directive 67/548/EEC, as amended for the 31st time in Directive 2009/2/EG.

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified for genetic toxicity under Regulation (EC) No 1272/2008, as amended for the sixth time in Regulation EC 605/2014.