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EC number: 700-748-4
CAS number: 1226911-69-8
7.4.1/1: Individual disintegrations per minute and stimulation indices –
main test – initial and additional test
Concentration (% v/v) in
acetone/olive oil 4:1
Disintegrations per minute
a = Total
number of lymph nodes per animal is 2
Stimulation Index of 3.0 or greater indicates a positive result
na = Not
Significantly different from control group p<0.01
= Significantly different from control group p<0.001
study was performed to assess the skin sensitisation potential of test
material in the CBA/Ca (CBA/CaOlaHsd) strain mouse following topical
application to the dorsal surface of the ear. The method was conducted
according to the OECD test guideline No 429 and in compliance with GLP.
a preliminary screening test in which no clinical signs of toxicity were
noted at a test item concentration of 100%, this concentration was
selected as the highest dose investigated in the main test of the Local
Lymph Node Assay. Three groups, each of five animals, were treated with
50 μl (25 μl per ear) of the undiluted test item or the test item as a
solution in acetone/olive oil 4:1 at concentrations of 50% or 25% v/v. A
further group of five animals was treated with acetone/olive oil 4:1
alone. A further group of five animals was treated with acetone/olive
oil 4:1 alone. In order to determine the concentration of test item
expected to cause a 3 fold increase in 3HTdR incorporation (EC3 value),
and at the request of the Study Sponsor, an additional group of five
animals was treated with the test item at a concentration of 5% v/v in
acetone/olive oil 4:1. An additional group of five animals was treated
with acetone/olive oil 4:1 alone.
were no deaths. No signs of systemic toxicity were noted in the test or
control animals during the test. No visual local skin irritation or
excessive irritation indicated by an equal to or greater than 25%
increase in mean ear thickness were noted at any dose concentration
evaluated. Bodyweight changes of the test animals between Day 1 and Day
6 were comparable to those observed in the corresponding control group
animals over the same period.
Stimulation Index values expressed as the mean radioactive incorporation
for each treatment group divided by the mean radioactive incorporation
of the vehicle control group are as follows: 2.12, 6.19, 13.93 and 15.06
at 5, 25 and 50% v/v in acetone/olive oil 4:1 and 100%, respectively. The
concentration of test item expected to cause a 3 fold increase in 3HTdR
incorporation (EC3 value) was calculated to be 9%.
historical positive control, α-Hexylcinnamaldehyde, gave a SI of 6.29,
when tested at 25 % v/v. The test system was therefore considered to be
the test conditions, test item is classified as a contact sensitiser
(Category 1B) according to the Regulation EC No. 1272/2008 (CLP) and to
study is considered as acceptable and satisfies the requirement for
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