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EC number: 215-125-8
CAS number: 1303-86-2
The study was performed according to the
OECD Guidelines for Testing of Chemicals No. 471 "Bacterial Reverse
Mutation Test", Method B13/14 of Commission Regulation (EC) number
440/2008 of 30 May 2008, the ICH S2(R1) guideline adopted June 2012 (ICH
S2(R1) Federal Register. Adopted 2012; 77:33748-33749) and the USA, EPA
OCSPP harmonized guideline - Bacterial Reverse Mutation Test in
compliance with GLP.
Salmonella typhimurium strains
TA1535, TA1537, TA98 and TA100 and Escherichia coli strain
WP2uvrA were treated with the test item using the Ames plate
incorporation method both with and without the addition of a
metabolizing system (10% liver S9 in standard co-factors). The dose
range for Experiment 1 was based on OECD TG 471 and was 1.5 to 5000
μg/plate. The dose range was amended following the results of Experiment
1 and was 15 to 5000 μg/plate.
The vehicle (sterile distilled water)
control plates gave counts of revertant colonies within the normal
range. All of the positive control chemicals used in the test induced
marked increases in the frequency of revertant colonies, both with and
without metabolic activation. There was no visible reduction in the
growth of the bacterial background lawn at any dose level, either in the
presence or absence of metabolic activation (S9-mix), in the first
mutation test. Similarly, there was no visible reduction in the growth
of the bacterial background lawn at any dose level, either in the
presence or absence of metabolic activation (S9-mix) in the second
No test item precipitate was observed on the
plates at any of the doses tested in either the presence or absence of
metabolic activation (S9-mix) in Experiments 1 and 2. There were no
significant increases in the frequency of revertant colonies recorded
for any of the bacterial strains, with any dose of the test item, either
with or without metabolic activation (S9-mix) in Experiment 1.
Similarly, no biologically relevant
increases in the frequency of revertant colonies were recorded for any
of the bacterial strains, with any dose of the test item, either with or
without metabolic activation (S9-mix) in Experiment 2. Minor statistical
values were noted in Experiment 2 (WP2uvrA at 15, 150 and 500 μg/plate
in the absence of S9-mix), however these responses were within the
in-house historical vehicle/untreated control ranges and were, therefore
considered of no biological relevance.
In conclusion, diboron trioxide (B2O3)
was considered to be non-mutagenic under the conditions of this test.
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