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Key value for chemical safety assessment

Additional information

There are reliable in vitro studies available to assess the potential of the test substance for gene mutations in bacteria and mammalian cells as well as cytogenicity in mammalian cells.

In vitro

Gene mutation in bacteria:

In a GLP conform study according to OECD guideline 471, the substance Golpanol ALS wasserfrei was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial strains, i.e. Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli WP2 uvrA (BASF SE, 2010g). Two independent assays were performed, a standard plate test (SPT) and a preincubation test (PIT), with a dose range of 20 µg - 5000 µg/plate for SPT and 312.5 - 5000 µg/plate for PIT, and both with and without metabolic activation (induced rat liver S-9 mix).

An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolizing system. No precipitation of the test item was found. A weak bacteriotoxic effect was occasionally observed depending on the strain and test conditions at 5 000 μg/plate. 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.

The described study above verifies the study from 1992 which is in accordance with "OECD principles of GLP" (1981). The substance Golpanol ALS there was tested for its mutagenic potential based on the ability to induce back mutations in selected loci in several strains of Salmonella typhimurium (TA 1535, TA 1537, TA 98 and TA 100) in the Ames test (BASF AG, 1992). Two independent assays were performed, a standard plate test (SPT) and a preincubation test (PIT), with a dose range of 2500 µg - 20000 µg/plate for both test types both with and without metabolic activation (induced rat liver S-9 mix). An increase in the number of his+ revertants was not observed both in the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

According to the results of the present studies, the test substance Golpanol ALS wasserfrei is not mutagenic in the Salmonella typhimurium/ Escherichia coli reverse mutation assay under the experimental conditions chosen here.

Gene mutation in mammalian cells:

In a GLP conform study according to OECD guideline 476, the substance Golpanol ALS wasserfrei was tested to investigate the potential of Golpanol ALS wasserfrei to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in V79 cells of the Chinese hamster (Harlan CCR GmbH). A pre-test was performed in order to determine the concentration range for the mutagenicity experiments. The assay was performed in two independent experiments, using two parallel cultures each. The maximum concentration of experiments I and II was again 7000 µg/mL since no relevant cytotoxic effects were expected. The first main 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.

No precipitation of the test item was observed up to the maximum concentration in all experimental parts.

No cytotoxic effects occurred up to the maximum concentration of 7000 µg/mL in both experiments with and without metabolic activation following 4 hours and 24 hours of exposure.

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test item and the activity of the metabolic activation system.

 

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

Therefore, Golpanol ALS wasserfrei is considered to be non-mutagenic in this HPRT assay.

Cytogenicity in mammalian cells:

In a GLP conform study according to OECD (Draft) guideline 487, the substance Golpanol ALS wasserfrei was assessed in Chinese Hamster V79 cells for possible clastogenic or aneugenic activity leading to inducement of micronuclei both in the presence or absence of a metabolizing system (BASF SE, 2010h). According to an initial range-finding cytotoxicity test for the determination of the experimental doses, the following doses were tested:

1st Experiment

4 hours exposure; 24 hours harvest time; without S9 mix: 0; 218.8; 437.5; 875.0; 1 750.0; 3 500.0; 7 000.0 μg/mL

4 hours exposure, 24 hours harvest time, with S9 mix: 0; 218.8; 437.5; 875.0; 1 750.0; 3 500.0; 7 000.0 μg/mL

2nd Experiment

24 hours exposure, 24 hours harvest time, without S9 mix: 0; 625.0; 1 250.0; 2 500.0; 3 500.0; 5 250.0; 7 000.0 μg/mL

4 hours exposure, 24 hours harvest time, with S9 mix: 0; 625.0; 1 250.0; 2 500.0; 3 500.0; 5 000.0; 7 000.0 μg/mL

A sample of at least 1 000 cells for each culture were analyzed for micronuclei, i.e. 2000 cells for each test group.

The vehicle controls gave frequencies of micronucleated cells within the available historical negative control data range for V79 cells. Both positive control substances and cyclophosphamide, led to the expected increase in the number of cells containing micronuclei. On the basis of the results of the present study, the test item did not cause any biologically relevant increase in the number of cells containing micronuclei either without S9 mix or after adding a metabolizing system. These findings were confirmed in two experiments carried out independently of each other.

 

Thus, under the experimental conditions described, Golpanol ALS wasserfrei is considered not to have a chromosome-damaging (clastogenic) effect nor to induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells in the absence and the presence of metabolic activation.


Short description of key information:
in vitro:
Gene mutation in bacteria:
Ames test, S. typhimurium/ E.coli, with and without metabolic activation: negative (GLP, OECD 471; BASF SE 2010g)

Gene mutation in mammalian cells:
HPRT test, V79 cells, with and without metabolic activation: negative (GLP, OECD 476; Harlan CCR GmbH 2010)

Cytogenicity in mammalian cells:
Micronucleus test, V79 cells, with and without metabolic activation: negative (GLP, OECD Draft 487; BASF SE 2010h)

in vivo:
no data

Endpoint Conclusion:

Justification for classification or non-classification

Based on the available in vitro studies, there is no indication for a mutagenic potential of the test substance.

Therefore, no classification is warranted for the genotoxic potential of the test substance according to the criteria of DSD (67/548/EEC) and CLP (1272/2008/EC), respectively.

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