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EC number: 283-829-2
CAS number: 84731-70-4
Ames test: negative (OECD 471);
Chromosome aberration test: negative (OECD 473);
In vitro gene mutation test: negative (OECD 490).
No contaminant colonies were observed on the sterility plates for the
highest concentration of the test substance or the S9 mix.
Confirmatory mutagenicity study:
There was no increase in the number of revertant colonies compared to
the vehicle control at any concentration of the test substance either in
the presence or absence of metabolic activation system in any strain
tested. The precipitation of test item was observed at the concentration
more than 250µg/plate
in all strains used, when test item solutions were mixed with top agar.
Precipitation was observed at the 500µg/plate
concentration after 48 h incubation. The number of viable cells in the
overnight cultures as determined by optical density was within
acceptable limits: 1.2 to 2.3E09 cells/mL (S. typhimurium strains) and
2.3E09 cells/mL (E. coli strain); 1.1 to 2.2E09 cells/mL (S. typhimurium
strains) and 2.1E09 cells/mL (E. coli strain) in the 1st and 2nd
experiments respectively. The positive controls induced a significant
increase in the number of revertant colonies indicating the assay was
Dosing solutions were analysed. The concentration analyses indicate that
the actual concentration of the dosing solutions used were 101.81 % to
103.82 % and 94.47 % to 103.81 % of target concentration in the 1st and
2nd experiments, respectively.
Mouse micronucleus test: negative (OECD 474).
Three in vitro genetic toxicity studies and an in vivo
micronucleus assay have been conducted on the test material, as follows:
Ames Test (in vitro):
The test item was evaluated for its potential to induce reverse mutation
using Salmonella typhimurium TA100, TA98, TA1535 and TA1537 and
Escherichia coli WP2uvrA in the presence or absence of metabolic
activation with S9 mix using the plate incorporation method. Under the
conditions of the study DEHCH was concluded to be negative. .
Chromosome aberration test (in vitro):
The potential of DEHCH to induce chromosomal aberrations in Chinese
Hamster Lung (CHL) cells in the presence or absence of metabolic
activation (S9 mix) was evaluated in the in vitro chromosome aberration
assay. Under the conditions of the study DEHCH did not induce
chromosomal aberrations in CHL cells.
In vitro gene mutation test:
This in vitro assay was performed to assess the potential of the test
item to induce mutations at the thymidine kinase locus of the mouse
lymphoma cell line L5178Y. In conclusion it can be stated that under the
experimental conditions reported the test item did not induce mutations
in the mouse lymphoma thymidine kinase locus assay using the cell line
L5178Y in the absence and presence of metabolic activation.
Mouse micronucleus assay ( in vivo)
A micronucelus test of DEHCH was performed using bone marrow cells of
specific pathogen free (SPF) 7 week old ICR mice The test item was
orally administered twice at 24 h intervals at doses of vehicle control,
500, 1000 and 2000 mg/kg in both male and female mice (6/sex/group). The
number of MNPCE (micronucleated polychromatic erythrocytes) was counted
in 2000 PCEs (polychromatic erythrocytes) per animal. There was no
statistically significant increase in the frequencies of MNCPEs at any
dose of DEHCH compared to the vehicle control. In addition no
significant difference was observed in the mean value for the ratio of
PCE to total erythrocytes (PCE/(PCE+NCE)) in the DEHCH group. DEHCH was
therefore concluded to be negative in the study.
Three in vitro genetic toxicity studies have been conducted on
the test material, an Ames study (OECD 471), a chromosome aberration
study (OECD 473), and in vitro gene mutation study (OECD 490) which were
both negative. An in vivo mouse micronuclues assay (OECD 474) is also
available which was also negative for mutagenicity.
All the in vitro and in vivo genetic toxicity studies showed the test
material to have no significant effects for gentoxicity. As such, the
test material can be considered to be non-classified.
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