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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Correlation between experimental human and murine skin sensitization induction thresholds
Author:
Anne Marie Api, David Basketter, and Jon Lalko
Year:
2015
Bibliographic source:
Cutaneous and ocular toxicology, 34, no. 4, (2015), 298-302.

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Principles of method if other than guideline:
To evaluate the skin sensitization potential of D-Carvone by Mouse Local Lymph Node Assay.
GLP compliance:
not specified
Type of study:
mouse local lymphnode assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material: d-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular Weight: 150.22 g/mole- Smiles Notation: C1[C@H](CC=C(C1=O)C)C(C)=C- Inchl: 1S/C10H14O/c1-7(2)9-5-4-8(3)10(11)6-9/h4,9H,1,5-6H2,2-3H3/t9-/m0/s1- Substance type: Organic- Physical state: Colorless to pale-yellow liquid
Specific details on test material used for the study:
- Name of test material: d-Carvone- IUPAC name: (5S)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one- Molecular formula: C10H14O- Molecular Weight: 150.22 g/mole- Smiles Notation: C1[C@H](CC=C(C1=O)C)C(C)=C- Inchl: 1S/C10H14O/c1-7(2)9-5-4-8(3)10(11)6-9/h4,9H,1,5-6H2,2-3H3/t9-/m0/s1- Substance type: Organic- Physical state: Colorless to pale-yellow liquid

In vivo test system

Test animals

Species:
mouse
Strain:
CBA/Ca
Sex:
female
Details on test animals and environmental conditions:
No data availlable

Study design: in vivo (LLNA)

Vehicle:
other: 1:3 ethanol :diethyl phthalate.
Concentration:
25 µl (Each group received one of five test concentrations prepared as a w/v%.)
No. of animals per dose:
Total=5
Details on study design:
MAIN STUDYANIMAL ASSIGNMENT AND TREATMENT- Name of test method: LLNA- Criteria used to consider a positive response: EC 3 value TREATMENT PREPARATION AND ADMINISTRATION: Groups of female CBA/CA mice were dosed topically on the dorsum of each ear with 25 µl of test fragrance in 1:3 ethanol: diethyl phthalate.Each group received one of five test concentrations preparedas a w/v%.
Positive control substance(s):
other:

Results and discussion

In vivo (LLNA)

Results
Key result
Parameter:
SI
Value:
2 650
Variability:
not specified
Test group / Remarks:
5 mice
Remarks on result:
other: Sensitizing to skin
Cellular proliferation data / Observations:
CELLULAR PROLIFERATION DATA: Suspensions of the lymph node cells were prepared by mechanical disaggregation through 200- mesh stainless steel gauze. The cell suspensions were washedthree times with phosphate buffered saline and precipitated overnight at 4°C with 5% w/v trichloroacetic acid (TCA). Thesamples were then pelleted by centrifugation. The cells wereresuspended in 1ml of TCA and transferred to scintillation vials containing 10 ml of scintillation fluid. The incorporationof 3HTdR was measured by b-scintillation counting and expressed as disintegrations per minute (dpm) per lymph node for each experimental group.DETAILS ON STIMULATION INDEX CALCULATION : The stimulation index (SI)values were calculated for each dose level. A SI of 3 or more was considered to give a positive response.EC3 CALCULATION : The EC3 valuewas taken as a measure of relative potency

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
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.
Executive summary:

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.