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EC number: 220-479-1
CAS number: 2781-00-2
The potential of the test item,
1,3(4)-bis(tert-butylperoxyisopropyl)benzene, to induce delayed contact
hypersensitivity using the murine Local Lymph Node Assay (LLNA) was
evaluated. Evaluation of local irritation was also carried out in
This study was conducted in compliance with
the principles of Good Laboratory Practice Regulations.
A preliminary test was first performed in
order to define the concentrations of test item to be used in the main
In the main test, twenty-eight female CBA/J
mice were allocated to seven groups:
. five treated groups of four animals
receiving the test item 1,3(4)-bis(tert-butylperoxyisopropyl)benzene at
the concentration of 2.5, 5, 10, 25 or 50% in a mixture acetone/olive
oil (4/1, v/v) (AOO),
. one negative control group of four animals
receiving the vehicle (AOO),
. one positivie control group of four
animals receiving the reference item, alpha-hexylcinnamaldehyde (HCA), a
moderate sensitizer, at the concentration of 25% in AOO.
During the induction phase, the test item,
vehicle or reference item was applied over the ears (25 µL per ear) for
3 consecutive days (days 1, 2 and 3). After 2 days of resting, the
proliferation of lymphocytes in the lymph node draining the application
site was measured by incorporation of tritiated methyl thymidine (day
6). The obtained values were used to calculate Stimulation Indices (SI).
The irritant potential of the test item was
assessed in parallel by measurement of ear thickness on days 1, 2, 3 and
The test item was soluble in the first
recommended vehicle, acetone/olive oil (4/1, v/v).
A solution was obtained at the maximum
tested concentration of 50%, after 15 minutes stirring.
Consequently, the concentrations selected
for the preliminary test were 5, 10, 25 and 50%.
Since the test item was non-irritant in the
preiminary test, the highest concentration retained for the main test
was the maximal practicable concentration (50%).
Systemic clinical signs and mortality
Neither mortality nor clinical signs were
observed during the study.
Dryness of the skin was noted on day 6 in
3/4 and 2/4 females given 25 and 50%, respectively.
No notable increase in ear thickness was
observed in the animals of the treated groups.
A significant lymphoproliferation was noted
with the positive control group given HCA at 25%. The study was
therefore considered valid.
No notable lymphoproliferation was noted at
any tested concentration.
The results are presented in the following
Under the experimental conditions of this
study, the test item 1,3(4)-bis(tert-butylperoxyisopropyl)benzene (batch
No. 307090713) did not induce delayed contact hypersensitivity in the
murine Local Lymph Node Assay.
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