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EC number: 470-080-3
CAS number: -
There are reliable in vitro studies
available to assess the potential of the test substance for both gene
mutations in bacteria and cytogenicity in mammalian cells.
Gene mutation in bacteria
In a GLP conform Ames test according to OECD
guideline 471, the substance
(purity: 93.0 weight-%) 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 2006). Two independent assays
were performed, a standard plate test (SPT) and a preincubation test
(PIT), both with a dose range of 56 µg - 5500 µg/plate, and both with
and without metabolic activation (Aroclor 1254-induced rat liver S-9
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. Precipitation of the test substance was found from
about 557 mg/plate onward. A slight decrease in the number of revertants
was occasionally observed depending on the strain and test conditions at
doses >= 1750 µg/plate.
According to the results of the present
study, the test substance is not mutagenic in the Salmonella
typhimurium/ Escherichia coli reverse mutation assay under the
experimental conditions chosen here.
Gene mutation in mammalian cells
In an in vitro gene mutation test (HPRT) in
CHO cells the substance
was assessed for its potential to induce gene mutations. Two independent
experiments were carried out, both with and without the addition of
liver S9 mix from induced rats (exogenous metabolic activation). The
following concentrations were assessed:
without S9 mix (4-hour exposure period)
0; 6.3;12.5; 25.0; 50.0; 75.0; 100.0; 150.0;
with S9 mix (4-hour exposure period)
0; 31.3; 62.5; 125.0; 500.0; 750.0; 1 000.0;
1 250.0; 1 500.0 μg/mL
without S9 mix (24-hour exposure period)
0; 1.6; 3.1; 6.3; 12.5; 20.0; 30.0; 40.0;
The vehicle controls gave mutant frequencies
within the range expected for the CHO cell line. Both positive control
substances, EMS and MCA, led to the expected increase in the frequencies
of forward mutations.
In the absence of metabolic activation, in
both experiments the highest concentrations applied could not be
evaluated for gene mutations due to pronounced cytotoxicity. In the
presence of metabolic activation, the highest concentration that could
be formulated in the vehicle DMSO (1500 μg/mL) led to slightly reduced
cloning efficiencies of about 50% survival in both experiments.
On the basis from the results of the present
study, the test substance did not cause any relevant increase in the
mutant frequencies neither without S9 mix nor after adding a
metabolizing system in two experiments performed independently of each
other, and apply clear cytotoxic or maximum soluble concentrations.
Thus, under the experimental conditions of
is not mutagenic in the HPRT locus assay under in vitro conditions in
CHO cells in the absence and the presence of metabolic activation.
Cytogenicity in mammalian cells
In a GLP conform study according to OECD
guideline 473, the substance
was assessed for its potential to induce structural chromosomal
aberrations (clastogenic activity) and/or changes in the number of
chromosomes (aneugenic activity) in V79 cells in vitro both in the
presence and in the absence of a metabolizing system.
According to an initial range-finding
cytotoxicity test for the determination of the experimental doses and
taking into account the cytotoxicity actually found in the main
experiments, the following doses were tested (the test groups in
brackets were not evaluated):
- 1st experiment:
4-hour exposure, 18-hour sampling time,
without S-9 mix: 0; 62.5 ; 125.0; 250.0 (500.0; 750.0; 1000) µg/mL;
4-hour exposure, 18-hour sampling time, with
S-9 mix: 0; (250.0;) 500.0 ; (750.0;) 1000.0; (1250.0;) 1500.0 µg/mL.
- 2nd experiment
18-hour exposure, 18-hour sampling time,
without S-9 mix: 0; 6.25; 12.5; 25.0; (50.0; 75.0; 100.0; 200.0) mg/mL
For confirmation of the results of the 1st
and 2nd experiments including continuous exposure and a second sampling
time, the following doses were tested in the amendment report:
- 3rd experiment
18-hour exposure, 28-hour sampling time,
without S-9 mix: 0; (6.3; 12.5; 25;) 50(; 100) μg/mL
4-hour exposure, 28-hour sampling time, with
S-9 mix: 0; 31.3; 62.5; (125;) 500; 1000(; 1500) μg/mL
About 2-3 hours prior to harvesting the
cells, Colcemid was added to arrest cells at a metaphase-like stage of
mitosis (c-metaphases). After preparation of the chromosomes and
staining with Giemsa, 100 metaphases for each culture in the case of the
test substance, and vehicle controls, or 50 cells for each culture in
the case of the concurrent positive controls, were analyzed for
The negative controls (vehicle controls)
gave frequencies of aberrations within the range expected for the V79
cell line. Both of the positive control chemicals, i.e. EMS and
cyclophosphamide, led to the expected increase in the number of cells
containing structural chromosomal aberrations.
On the basis of the results of the present
study, the test substance did not cause any increase in the number of
structurally aberrant metaphases incl . and excl. gaps either without
S-9 mix or after adding a metabolizing system in two experiments
performed ndependently of each other and apply clear cytotoxic
concentrations. No increase in the frequency of polyploid cells was
demonstrated either. Thus, under the experimental conditions of this
assay, the test substance is considered not to be either a clastogenic
or an aneugenic agent under in vitro conditions in V79 cells.
Based on the available in vitro studies,
there is no indication for a mutagenic potential of the test substance. Thus,
the test substance has not to be classified with regard to mutagenicity according
to 67/548/EEC and to Regulation (EC) No 127272008 (GHS, CLP).
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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