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EC number: 245-844-2
CAS number: 23726-93-4
No irritation or other adverse toxic effects were noted in any of the
mice used in this study, and there were no animal deaths in any of the
- There was some weight loss (0.1-1.1 g) was
observed in 11/48 animals over the 6 day period from dosing to lymph
node harvest. This weight loss occurred across dose and treatment
groups. The animals appeared healthy otherwise with a final body weight
range of 19.8-22.9 g
The highest doses (1, 2.5 and 5 %) of the test article had SI of 3.5,
6.2 and 7.9. A one-sample t test on the untransformed SI values
indicated that the SI value for 2.5 and 5 % were statistically
significant. All the other doses tested (0.25 and 0.5 %) had SI values
less than 3.0.
In a Local
Lymph Node Assay (LLNA) conducted similarly to the OECD test guideline
No 429 and in compliance with GLP, groups of female CBA/J mice (5
females/group) were topically applied with test material at the dose
concentrations of 0.25, 0.5, 1.0, 2.5 and 5.0 % final concentration in
1:4 acetone:olive oil to the dorsal surface of both ears (25 µL/ear)
daily for three consecutive days. A vehicle control group (8
females/group) was treated with 1:4 acetone:olive oil alone and a
positive control group (5 females/group) was treated with isoeugenol at
the dose concentration of 0.5, 1.0 and 5.0 % in acetone:olive oil (4:1)
in same manner to confirm the sensitivity and reliability of the test
method. The animals were allowed to rest without dosing on Days 4 and 5.
On Day 6, animals were injected intravenously with 125I- labelled luDR
to label proliferating cells. 125I-incorporation was quantified using a
gamma counter and disintegrations per minute (DPM) and stimulation index
(Sl) were calculated for each dose group.
positive control, Isoeugenol gave SI of 4.2 and 18.4, when tested at 1
and 5 % in 4:1 acetone/olive oil, respectively. The test system was
therefore considered to be valid.
mortality, irritation or other adverse toxic effects were noted in any
of the animals. Mean DPM for 0 (vehicle), 0.25, 0.5, 1.0, 2.5 and 5.0 %
were 26.9, 16.9, 42.8, 95.2, 167.6 and 213.9, respectively. Stimulation
Index (SI Value) calculated for test material was found to be 0.6, 1.6,
3.5, 6.2 and 7.9 for the dose concentrations of 0.25, 0.5, 1.0, 2.5 and
5.0 %, respectively. The highest doses (1, 2.5 and 5 %) of the test
article had SI of 3.5, 6.2 and 7.9. A one-sample t test on the
untransformed SI values indicated that the SI value for 2.5 and 5 % were
statistically significant. All the other doses tested (0.25 and 0.5 %)
had SI values less than 3.0. The calculated EC-3 for test material using
a fitted quadratic equation was 1.22% and potency value was 305 µg/cm2.
test conditions, test material is classified asa skin sensitiser
according to the annex VI of the Regulation EC No. 1272/2008 (CLP).
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