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EC number: 218-827-2
CAS number: 2244-16-8
Skin sensitization potential of d-Carvone was evaluated by Local Lymph Node Assay (LLNA) The LLNAs were conducted according to the method described in OECD Guideline 429. Groups of female CBA/CA mice (n= 5) were dosed topically on the dorsum of each ear with 25 ml of test material in 1:3 ethanol :diethyl phthalate. Each group received one of five test concentrations prepared as a w/v%. A concurrent vehicle control group was similarly treated with 1:3 ethanol: diethyl phthalate.
Dosing occurred daily for three consecutive days. The animals received no treatment for two days and on the sixth day after the first application, all mice were injected intravenously via the tail vein with 250 ml of phosphate buffered saline containing 20 µCi of radiolabelled methyl thymidine (3HTdR). Five hours later, the mice were euthanized and the draining auricular lymph nodes were excised and pooled . Suspensions of the lymph node cells were prepared by mechanical disaggregation through 200-mesh stainless steel gauze. The cell suspensions were washed three times with phosphate buffered saline and precipitated overnight at 4 °C with 5% w/v trichloroacetic acid (TCA). The samples were then pelleted by centrifugation. The cells were resuspended in 1ml of TCA and transferred to scintillation vials containing 10 ml of scintillation fluid. The incorporation of 3HTdR was measured by β- scintillationcounting and expressed as disintegrations per minute (dpm) per lymph node.
The EC3 value obtained from LLNA assay performed to assess the sensitizing potential of d-Carvone was 2650 microgram/sq cm. Based on this value, d-Carvone can be considered to be sensitizing to skin.
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