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EC number: 248-698-8
CAS number: 27859-58-1
Table1. Summary of Results
MF threshold for a positive response
%RSG = Relative Suspension Growth
RTG = Relative Total Growth
MF§ = 5-TFT resistant mutants/106 viable cells 2 days after treatment
CP = Cyclophosphamide
EMS = Ethylmethanesulphonate
Ø = Not plated for viability or 5-TFT resistance
The mutagenic activity of the test material was evaluated in an in vitro
mammalian cell gene mutation test with L5178Y mouse lymphoma cells. The
GLP study was conducted in accordance with the standardised guidelines
OECD 490 and EU Method B.17, US EPA OPPTS 870.5300 and in alignment with
the Japanese MITI/MHW guidelines for testing of new chemical substances.
One main Mutagenicity Test was performed and in this main test, L5178Y
TK +/- 3.7.2c mouse lymphoma cells (heterozygous at the thymidine kinase
locus) were treated with the test material at 8 dose levels in
duplicate, together with vehicle (R0 media), and positive controls using
4-hour exposure groups both in the absence and presence of metabolic
activation (2 % S9), and a 24-hour exposure group in the absence of
metabolic activation. The dose range of test material used in the main
test was selected following the results of a preliminary toxicity test.
The test material exhibited dose-related toxicity to the cells in each
of the three exposure groups of the preliminary toxicity test. The dose
levels plated for viability and expression of mutant colonies were 3.91,
7.81, 15.63, 31.25, 62.5 and 125 µg/mL for 4-hour treatment without S9,
4-hour treatment with S9 (2 %) and 24-hour treatment without S9. The
maximum dose level used was limited by the presence of precipitate. A
cloudy precipitate of test material was observed at and above 62.5 µg/mL
at the end of the exposure period in all three exposure groups. The
vehicle control cultures had mutant frequency values that were
acceptable for the L5178Y cell line at the TK +/- locus. The positive
control substances induced marked increases in the mutant frequency,
sufficient to indicate the satisfactory performance of the test and of
the activity of the metabolizing system. The test material did not
induce any toxicologically significant or dose-related (linear-trend)
increases in the mutant frequency at any of the dose levels in the main
test, in any of the three exposure groups.
Under the conditions of the test, the test material did not induce any
increases in the mutant frequency at the TK +/- locus in L5178Y cells
that exceeded the Global Evaluation Factor (GEF) of 126 x 10^-6,
consequently it is considered to be non-mutagenic in this assay
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