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EC number: 227-815-6
CAS number: 5989-54-8
Table 1: Local lymph node assay responses to
Exposure Concentration (% v/v)
Stimulation index (SI)
In a skin sensitization study conducted
following a method similar to OECD guidelines 429, four groups of CBA/Ca
strain female mice (4/concentration) were exposed topically on the
dorsum of both ears to 25 µL of vehicle (acetone/olive oil, 4:1 v/v),
25, 50 and 100% v/v of d-limonene daily for 3 consecutive days. Five
days following initiation of exposure all mice were injected 250 µL of
phosphate buffered saline (PBS) containing 20 µCi of [3H] methyl
thymidine (3H-TdR). The animals were then sacrificed after 5 hours and
suspension of lymph node cells was prepared from auricular lymph nodes.
The incorporation of 3H-TdR was measured by β-scintillation counting as
disintegrations per minute (dpm) per node for each experimental group. A
Stimulation index of 3 or greater was considered to be indicative of a
potential to cause contact sensitization.
D-limonene at exposure concentration of 0
(vehicle), 25, 50 and 100% v/v resulted in 476, 877, 1164 and 1882
dpm/node and stimulation index of 1, 1.84, 2.44 and 3.95, respectively.
The calculated EC3-value for d-limonene was
found to be 68.5%. Under these test conditions, d-limonene was found to
be sensitising and should be clasified as skin sensitiser according to
the CLP Regulation (EC) N° (1272-2008).
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