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EC number: 248-256-4
CAS number: 27138-01-8
The objective of this study was to
evaluate the potential of the test item to induce reverse mutation in Salmonella
The study was performed according to
the international guidelines (OECD No. 471 and Commission Directive No.
B.13/14) and in compliance with the principles of Good Laboratory
A preliminary toxicity test was
performed to define the dose-levels of m/p DIOL to be used for the
mutagenicity study. The test item was then tested in two independent
experiments, with and without a metabolic activation system, the S9 mix,
prepared from a liver post-mitochondrial fraction (S9 fraction) of rats
induced with Aroclor 1254.
Both experiments were performed
according to the direct plate incorporation method except for the second
test with S9 mix, which was performed according to the pre-incubation
method (60 minutes, 37°C).
Five strains of bacteria Salmonella
typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 102 were used.
Each strain was exposed to at least five dose-levels of the test item
(three plates/dose-level). After 48 to 72 hours of incubation at, the
revertant colonies were scored.
The evaluation of the toxicity was
performed on the basis of the observation of the decrease in the number
of revertant colonies and/or a thinning of the bacterial lawn.
The test item was dissolved in
The number of revertants for the
vehicle and positive controls was as specified in the acceptance
criteria. The study was therefore considered to be valid.
Since the test item was found freely
soluble and non-cytotoxic in the preliminary test, the highest
dose-level was 5000 µg/plate, according to the criteria specified in the
The selected treatment-levels were:
312.5, 625, 1250, 2500 and 5000 µg/plate for both mutagenicity
experiments, with and without S9 mix.
No precipitate was observed in the
Petri plates when scoring the revertants at any dose-levels.
No noteworthy toxicity (decrease in
the number of revertants or thinning of the bacterial lawn) was noted
towards any of the tested strains, either with or without S9 mix.
No noteworthy increase in the number
of revertants, which could be considered as biologically relevant, was
noted in any of the five strains used, either with or without S9 mix.
The test item did not show any
mutagenic activity in the bacterial reverse mutation test with Salmonella
typhimurium, either in the presence or in the absence of a rat liver
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