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EC number: 700-957-0
CAS number: -
No systems of local toxicity at the ears of the animals and no systemic
findings were observed during the study period.
Table 1. Calculation of individual DPM and SI values
Test substance concentr.
DPM values measured
DPM-BG per animal (2 lymph nodes)*
13 (BG I)
12 (BG II)
BG = background (1 mL 5% trichloroacetic acid)
1 = control group; 2-4: test groups
SI = stimulation index relative to the mean of the control group (group
* values corrected for mean background value (BG I and BG II)
Table 2. Calculation of mean DPM and SI values
Test substance concentration
DPM per lymph node#
# DPM/node was determined by dividing the sum of the measured values
from all lymph nodes within a group by the number of lymph nodes taken
from that group.
*Statistically significant compared to control (p = 0.005)
Table 3. Calculation of the EC3 value
EC3 = (a-c) [(3-d)/(b-d)] + c = 59.9 %
EC3 = estimated concentration for a SI of 3
a, b, c, d = co-ordinates for the 2 pairs of data lying immediately
above and below the SI value of 3 on the LLNA dose response plot
The skin sensitising potential of Y-15866 was investigated in a
Local Lymph Node Assay (LLNA) in mice according to OECD guideline 429
and in compliance with GLP. Based on a preliminary test in two female
CBA/CaOlaHsd mice, the undiluted test substance (100%) and dilutions of
25 and 50% (w/v) of the test substance in dimethylformamide (DMF) were
selected as treatment concentrations for the main study. In this
experiment, 5 female CBA/CaOlaHsd mice per test group were treated with
the undiluted or diluted test substance or vehicle alone, respectively.
The test substance formulations or the vehicle were applied on the
external surface of each ear (25 µL/ear) for three consecutive days.
Five days after the first topical application, the cell proliferation of
pooled lymph nodes from individual animals was measured by incorporation
of ³H-methyl thymidine and expressed as the amount of radioactive
disintegration per minute (DPM). The mean DPM/lymph node for each test
group was 520.8, 784.6 and 1450.2 at concentrations of 25, 50 and 100%
of the test substance, respectively. Treatment with the undiluted test
substance (100% concentration) resulted in a statistically significant
increase in DPM/lymph node (1450.2) compared to control values (DPM/node
= 305.3) and showed a clear dose-response relationship. Based on these
results, stimulation indices of 4.75, 2.57 and 1.71 were calculated for
the treatment concentrations of 100%, 50% and 25%, respectively. The
estimated concentration for a stimulation index of 3 (EC3) was 59.9%. No
local or systemic toxicity and no effects on body weights were observed.
The historical positive control hexyl cinnamic aldehyde confirmed the
sensitivity and reliability of the experimental technique (SI ≥ 3).
Under the above mentioned conditions, the test substance was found to be
a sensitiser in the LLNA.
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