1-Butanamine, N-[3-(trimethoxysilyl)propyl]-, reaction products with (trimethoxysilyl)-N-[(trimethoxysilyl)propyl]-1-propanamine and 1,6-diisocyanatohexane homopolymer

Endpoint summary

Additional information

in vitro:

Ames-Test

iGloss Crosslinker (ZQ54-2211) was tested in a GLP compliant Ames reverse mutation assay according to OECD guideline 471 using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100 and Escherichia coli WP2 uvr A at 33 to 5000 µg/plate (two independent experiments (standard plate incorporation test and pre-incubation test) and each concentration was tested in triplicate) with and without metabolic activation (BASF SE, 2012). Precipitation of the test substance was found at 5000 μg/plate with and without S9 mix. A bacteriotoxic effect was observed depending on the strain and test conditions from about 2500 μg/plate onward. A relevant increase in the number of his⁺or trp⁺revertants was not observed in the standard plate test or in the preincubation test either without S9 mix or after the addition of a metabolizing system. Thus, under the experimental conditions of this study, the test substance iGloss Crosslinker (ZQ54-2211) is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay in the absence and the presence of metabolic activation.

Chromosome Aberration Test:

iGloss Crosslinker (ZQ54-2211) was tested in a GLP compliant in vitro mammalian chromosome aberration test according to OECD guideline 473 in which Chinese hamster V79 cells were exposed to 19.5 to 5000 (IA), 0.8 to 200 (IB) or 5 to 100 (IC) µg/mL without metabolic activation and 19.5 to 5000 (IA) or 10 to 200 (IC) µg/mL with metabolic activation (Harlan CCR, 2012).

In Experiment IA and IB in the absence or presence of S9 mix concentrations showing clear cytotoxicity were not scorable for cytogenetic damage. In Experiment IC clear cytotoxicity indicated by reduced cell numbers or mitotic indices was observed at the highest evaluated concentrations. In the absence of S9 mix no clastogenicity was observed at the concentrations evaluated. In Experiment IA in the presence of S9 mix one statistically significant increase in chromosomal aberrations (6.0 % aberrant cells, excluding gaps) clearly exceeding the historical solvent control data (0.0 – 4.0 % aberrant cells, excluding gaps) was observed after treatment with 78.1 µg/mL. In the confirmatory experiment IC one statistically significant increase in chromosomal aberrations (7.5 % aberrant cells, excluding gaps) clearly exceeding the historical solvent control data (0.0 – 4.0 % aberrant cells, excluding gaps) was observed after treatment with 80.0 µg/mL. No evidence of an increase in polyploid metaphases was found after treatment with the test item as compared to the frequencies of the control cultures. Appropriate mutagens (EMS and CPA) were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations. In conclusion, it can be stated that under the experimental conditions reported, the test item induced structural chromosome aberrations in V79 cells (Chinese hamster cell line) in vitro. Therefore, iGloss Crosslinker (ZQ54-2211) is considered to be clastogenic in this chromosome aberration test in the presence of metabolic activation, when tested up to cytotoxic, precipitating or the highest evaluable concentration.

HPRT-Test:

A GLP-compliant gene mutation assay, tested according to OECD guideline 476, was performed to investigate the potential of iGloss Crosslinker (ZQ54-2211) to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster (Harlan 2012). The assay was performed in two independent experiments, using two parallel cultures each. The first experiment was performed with and without liver microsomal activation and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.The concentration range of the main experiments was limited by the cytotoxicity and precipitation of the test item. Relevant cytotoxic effects were observed in the first experiment at 10.0μg/mL and above without metabolic activation. In the presence of metabolic activation no cytotoxic effects were noted up to the second highest concentration of 80.0μg/mL. At the maximum concentration of 160.0μg/mL exceedingly severe cytotoxicity precluded evaluation of any results. In the second experiment cytotoxic effects as described above were noted at 80.0μg/mL with and without metabolic activation. The recommended cytotoxic range of approximately 10-20 % relative cloning efficiency I or relative cell density was covered with and without metabolic activation. No substantial and reproducible dose dependent increase of the mutation frequency was observed up to the maximum concentration with and without metabolic activation. Therefore, iGloss Crosslinker (ZQ54-2211) is considered to be non-mutagenic in this HPRT assay.

in vivo:

Micronucleus Assay

This study was performed to investigate the potential of iGloss Crosslinker (ZQ54-2211) to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse according to OECD 474 guideline and GLP (Harlan, 2013).

The test item was dissolved in 30% DMSO and 70% PEG 400, which was also used as vehicle control. The volume administered orally was 10 mL/kg b.w.. 24 h and 48 h after a single administration of the test item the bone marrow cells were collected for micronuclei analysis.

Seven males per test group (except the vehicle and positive control groups with 5 males only) were evaluated for the occurrence of micronuclei. Per animal 2000 polychromatic erythrocytes (PCEs) were scored for micronuclei.

To investigate a cytotoxic effect due to the treatment with the test item the ratio between polychromatic and normochromatic erythrocytes was determined in the same sample and reported as the number of PCEs per 2000 erythrocytes.

The following dose levels of the test item were investigated, based on results of a pre-experiment:

24 h preparation interval: 500, 1000, and 2000 mg/kg b.w.
48 h preparation interval: 2000 mg/kg b.w.

After treatment with the test item the number of polychromatic erythrocytes (PCEs) per 2000 erythrocytes was not substantially decreased as compared to the mean value of PCEs per 2000 erythrocytes of the vehicle control thus indicating that iGloss Crosslinker (ZQ54-2211) did not exert a cytotoxic effect in the bone marrow.

In comparison to the corresponding vehicle controls there was no biologically relevant or statistically significant enhancement in the frequency of the detected micronuclei at any preparation interval after administration of the test item and with any dose level used.

40 mg/kg b.w. cyclophosphamide administered orally was used as positive control which showed a substantial increase of induced micronucleus frequency

In conclusion, it can be stated that under the experimental conditions reported, the test item iGloss Crosslinker (ZQ54-2211) did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse.

Therefore, iGloss Crosslinker (ZQ54-2211) is considered to be non-mutagenic in this micronucleus assay.

Short description of key information:
in vitro:
Ames-Test (BASF, 2012): negative
HPRT-Assay (Harlan, 2012): negative
in vivo
Mikronucleus-Assay (Harlan, 2013): negative

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Negative results were obtained with an Ames-Test and with a HPRT-Test. The positive in vitro results in a chromosome aberration test could not be confirmed in an in vivo Micronucleus Assay. Therefore no classification for mutagenicity is required according to EU directive 67/548/EEC and EU classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.