L-p-mentha-1(6),8-dien-2-one

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01-09-99 to 22-09-99

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

other company data

Title:

Unnamed

Year:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity) and Method B.3 of Commission Directive 92/69/EEC

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

L-p-mentha-1(6),8-dien-2-one

EC Number:

229-352-5

EC Name:

L-p-mentha-1(6),8-dien-2-one

Cas Number:

6485-40-1

Molecular formula:

C10H14O

IUPAC Name:

(5R)-2-methyl-5-(prop-1-en-2-yl)cyclohex-2-en-1-one

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): CARVON L SYNTH
- Physical state: liquid
- Analytical purity: 99.4%
- Lot/batch No.: 50458154
- Storage condition of test material: approximately 4 °C in the dark
- Other: For the purpose of the study the test material was used as supplied. The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.

Test animals

Species:

rat

Strain:

other: Sprague-Dawley CD (Crl : CD (SD) IGS BR)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: Approximately eight to twelve weeks old
- Weight at study initiation: Male - 226 to 241 g; Female - 216 to 233 g.

- Housing: The animals were housed in suspended polypropylene cages furnished with woodflakes. The animals were housed individually during the 24-hour exposure period and in groups of five, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): Rat and Mouse SQC Expanded Diet No.1, Special Diets Services Limited, Witham, Essex, UK ad libitum
- Water (e.g. ad libitum): Mains drinking water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Within 19 to 25°C target range
- Humidity (%): Within 30 to 70% target range
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Back and flanks of each animal were clipped free of hair. The substance was applied uniformly to an area of shorn skin.
- % coverage: Approximating to 10% of the total body surface area
- Type of wrap if used: Surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair was wiped with cotton wool moistened with distilled water to remove any residual test material.
- Time after start of exposure: After 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.10 mL/kg (2000 mg/kg; specific gravity: 0.956)

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 male and 5 female

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Observed for deaths or overt signs of toxicity: 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.
Observations for evidence of primary irritation: Once daily every 14 days.
Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.

- Necropsy of survivors performed: yes

- Other examinations performed: clinical signs, body weight, dermal reactions.
The dermal reactions were numerically graded daily for erythema and eschar formation [0 (no erythema) to 4 (severe erythema to slight eschar formation)] and edema [0 (no edema) to 4 (severe edema); see Draize scoring system below].

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

There were no deaths (Table 1).

Clinical signs:

other: No clinical signs of toxicity were noted during the study (Table 1).

Gross pathology:

No abnormal macroscopic findings at necropsy.

Other findings:

- Other observations: No signs of dermal irritation were noted during the study (Table 2; Draize scoring system) .

Any other information on results incl. tables

Table 1: INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA

Dose Level  mg/kg	Animal Number and Sex	Effects Noted After Dosing (Hours)			Effects Noted During Period After Dosing (Days)
2000 mg/kg		0.5	1	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
	Male 1-0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Male 1-1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Male 1-2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Male 1-3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Male 1-4	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Female 2-0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Fernale 2-1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Fernale 2-2	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Female 2-3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Female 2-4	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

0 = no signs of systemic toxicity

Study report attachments:

Table 2 Individual dermal reactions (012_300)

Table 3 Individual bodyweights and weekly bodyweight changes (012_300)

Applicant's summary and conclusion

Interpretation of results:

not classified

Conclusions:

The acute dermal median lethal dose (LD50) of L-carvone in the Sprague-Dawley CD strain rat was found to be greater than 2000 mg/kg bw.

Executive summary:

In an acute dermal toxicity study (012_300), young adult Sprague-Dawley CD (Crl : CD (SD) IGS BR) rats (5/sex) were dermally exposed (semi-occlusive; approximately 10% of the total body surface area) to L-carvone for 24 hours in a limit test at a dose of 2000 mg/kg bw. Animals were then observed for 14 days.

The dermal LD50 Male/Female was greater than 2000 mg/kg bw (limit test). There were no treatment related clinical signs, necropsy findings, changes in body weight or signs of dermal irritation noted during the study.

This acute dermal study is classified as acceptable. It does satisfy the guideline requirement for an acute dermal study (OECD 402) in the rat.