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EC number: 245-844-2
CAS number: 23726-93-4
7.6.1/2: Preliminary toxicity Test
WP2 uvr A
a reverse gene mutation assay performed according to the OECD test
guideline No. 471 and in compliance with GLP, strains of Salmonella
typhimurium (TA 1535, TA 1537, TA 98 and TA 100) and Escherichia
coli WP2 uvr A were exposed to the test material diluted in DMSO at
the following concentrations using the plate incorporation method. The
dose range for the range-finding test was determined in a preliminary
toxicity assay and was 5 to 5000 and 50 to 5000 µg/plate for the Salmonella
strains and E.coli strain WP2uvrA- respectively. The
experiment was repeated on a separate day using a slightly amended dose
range (15 to 5000 and 50 to 5000 µg/plate for the Salmonella strains
and E.coli strain WP2uvrA- respectively), fresh cultures of the
bacterial strains and fresh test material formulations. Experiment was
carried out in
triplicate, both with and without metabolic activation (10 % liver S9 in
standard co-factors). Negative,
vehicle (DMSO) and positive control groups were also included in
vehicle 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 or without metabolic activation. Thus, the sensitivity of the assay
and the efficacy of the S9-mix were validated.
test material caused a visible reduction in the growth of the bacterial
background lawn and/or a substantial decrease in revertant colony
frequency to all of the Salmonella strains, initially at 1500
µg/plate both in the presence and absence of S9. However no toxicity was
noted to E.coli strain WP2uvrA- at any test material dose level. These
results were not indicative of toxicity sufficiently severe enough to
prevent the test material being tested up to the maximum recommended
dose level of 5000 µg/plate.
A light (oily in appearance) precipitate was observed at and above 1500
µg/plate, this did not prevent the scoring of revertant colonies. No
significant increases in the frequency of revertant colonies were
recorded for any of the bacterial strains, at any dose level either with
or without metabolic activation.
the test condition, the test material is not mutagenic with and without
metabolic activation to S. typhimurium (TA 1535, TA 1537, TA 98
and TA 100) and E. coli WP2 uvr A according to the criteria of
the Annex VI of the Regulation (EC) No. 1272/2008 (CLP).
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