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EC number: 204-279-1
CAS number: 118-82-1
The test substance AN-2 was examined for its potential to induce
structural chromosomal aberrations in Chinese Hamster Ovary (CHO) cells,
in both the absence and presence of a metabolic activation system
(S9-mix). Dimethylsulfoxide (DMSO) was used as vehicle for the test
substance. Two chromosomal aberration tests were conducted. In all
instances, duplicate cultures were used. Dose levels, ranging from 1.25
to 300 micro-g/ml, were tested. Both solubility and cytotoxicity were
used for dose level selection. Mitotic index reduction was used as
indication for cell toxicity.
In the first chromosomal aberration test, in both the absence and
presence of a metabolic activation system (S9-mix). treatment/harvesting
time was 4/18 hours (pulse treatment) and three concentrations of the
test substance (50, 100 and 200 micro-g/ml) were selected for
chromosomal aberration analysis. In the pulse treatment group with
metabolic activation (S9-mix), the highest concentration was weakly
cytotoxic to the cells. In the pulse treatment group without metabolic
activation (S9- mix), the two highest concentrations were clearly
cytotoxic to the cells.
In the second chromosomal aberration test, in the presence of S9-mix.
treatment/harvesting time was 4/18 hours (pulse treatment) and three
concentrations of the test substance (100, 200 and 300 micro-g/ml) were
selected for chromosomal aberration analysis. Dose-related cytotoxicity
was induced by the test substance. In the absence of S9-mix,
treatment/harvest time was 18/18 hours (continuous treatment) and three
concentrations of the test substance (2.5, 5 and 10 micro-g/ml) were
selected for chromosomal aberration analysis. The test substance was
weakly cytotoxic to the cells at the two lowest concentrations and
clearly cytotoxic to the cells at the highest concentration. At the next
higher concentration (30 micro-g/ml), the test substance was too
cytotoxic and therefore not selected for chromosomal aberration analysis.
In both the first and second chromosomal aberration lest, the test
substance AN-2 did not induce a statistically significant increase in
the number of aberrant cells, at any of the concentrations and treatment
periods analysed. when compared to the number of aberrant cells found in
the vehicle (DMSO) control cultures. In both chromosomal aberration
tests, the numbers of aberrant cells. found in the vehicle (DMSO)
control cultures. were within the historical range and the positive
control substances mitomycin C (in the absence of the S9·mix) and
cyclophosphamide (in the presence of the S9-mix) induced the expected
statistically significant increases in the incidence of structural
chromosomal aberrations. This demonstrates the validity of the assay.
Under the conditions used in this study. the test substance AN-2 was not
clastogenic to Chinese Hamster Ovary (CHO) cells.
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