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EC number: 200-262-8 | CAS number: 56-23-5
The potential of the test item, CARBON TETRACHLORIDE, to induce delayed contact hypersensitivity using the murine Local Lymph Node Assay (LLNA) was evaluated. Evaluation of local irritation was also carried out in parallel.
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 test.
In the main test, 20 female CBA/J mice were allocated to five groups:
. three treated groups of four animals each receiving the test item at the concentrations of 25%, 50% or 100% in a mixture of acetone/olive oil (4/1; v/v) (vehicle),
. one negative control group of four animals receiving the vehicle,
. one positive control group of four animals receiving the reference item, alpha-hexylcinnamaldehyde (HCA), a moderate sensitizer, at the concentration of 25% the vehicle.
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 6.
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%.
Consequently, the concentrations selected for the preliminary test were 10%, 25%, 50% and 100%.
Since the test item was non-irritant in the preliminary test, the highest concentration retained for the main test was the maximal practicable concentration (100%).
No treatment-related mortality or clinical signs were observed during the main test.
Erythema was noted on day 6 in 1/4 females given 100%. No notable increase in ear thickness was observed in any treated animal.
A significant lymphoproliferation was noted in the positive control group given HCA at 25%. The study was therefore considered valid.
A dose-related increase in the SI was noted at all the concentrations and the threshold of 3 was exceeded at the concentration of 100%.
The results are presented in the following table:
In the absence of local irritation, the positive lymphoproliferative response observed was attributed to delayed contact hypersensitivity. The EC3 value for the test item, CARBON TETRACHLORIDE, is equal to 58%. Therefore, on the basis of relative skin sensitization potency, the test item was classified as weak sensitizer. Conclusion Under the experimental conditions of this study, the test item, CARBON TETRACHLORIDE, (batch No. R864 du 30/11/2009), demonstrated weak dermal sensitization potential in the murine Local Lymph Node Assay and thus is classified skin sensitizer category 1B according to CLP regulation EU No 286/2011.
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