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Description of key information

LD50 Oral (male/female): 5400mg/kg bw  (OECD 401;GLP)  

LD50 Dermal: >2000mg/kg bw  (OECD 402; GLP)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 May 1986 - 26 June 1986
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: Annex V 79/831/EEC Method B.1; OECD Guideline 401
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: CFY (Sprague Dawley origin)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Interfauna U.K. Ltd., Huntington, Cambridgshire, UK.
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 90-142g
- Housing: Within treatment groups, by sex, in metal cages with wire mesh floors.
- Diet (e.g. ad libitum): A standard rodent diet (Labsure LAD 1 diet) ad libitum.
- Fasting period before study: Access to food only was prevented overnight prior to and approx. 4 hours after dosing.
- Water (e.g. ad libitum): Drinking water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Mean daily minimum and maximum were 21°C and 23°C
- Humidity (%): Mean daily relative humidity was 59%
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours aritifical light in each 24 hour period
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED:
3.30 mL/kg : 3.2 g/kg dose
5.15 mL/kg : 5.0 g/kg dose
8.25 mL/kg : 8.0 g/kg

Doses:
Preliminary test: single dose of 5.0 g/kg bw of undiluted test substance

Main test: 3.2, 5.0, or 8.0 g/kg of undiluted test substance at a dose volume of 3.30, 5.15, or 8.25 mL/kg respectively.
No. of animals per sex per dose:
Preliminary test: 2 male and 2 female animals

Main test: 5 male and 5 female animals
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Observations: Soon after dosing, frequently on Day 1, on subsequent days once in the morning and the evening; 5 and 14 days observations for preliminary and main tests respectively.
Body weights were recorded on Days 1 (day of dosing), 8, and 15.

- Necropsy of survivors performed: yes

- Other examinations performed: clinical signs, body weight, terminal autopsy
Statistics:
The acute median lethal oral dose was calculated by method of Weil.
(Weil, C.S. (1952) Biometrics, 8: 249.)
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 400 mg/kg bw
95% CL:
4 600 - 6 300
Sex:
male
Dose descriptor:
LD50
Effect level:
5 900 mg/kg bw
95% CL:
4 900 - 7 100
Sex:
female
Dose descriptor:
LD50
Effect level:
4 900 mg/kg bw
95% CL:
3 900 - 6 100
Mortality:
Preliminary test (males and females):
At 5 g/kg bodyweight: 0/4 deaths;


Main test (males):
At 3.2 g/kg: 0/5 deaths;
At 5 g/kg: 1/5 deaths (by day 4);
At 8 g/kg: 5/5 deaths (by day 3).

Main test (females):
At 3.2 g/kg: 0/5 deaths;
At 5 g/kg: 3/5 deaths (by day 4);
At 8 g/kg: 5/5 deaths (by day 5).
Clinical signs:
other: All rats: piloerection, hunched posture, increased salivation; abnormal gait (waddling) in majority of rats; isolated cases of prostration and straub tail. At 3.2 g/kg: lethargy (one male), pallor of extremities, ataxia and body tremors; clinical signs
Gross pathology:
All rats surviving to the end of the observation period were killed on day 15 by cervical dislocation and examined macroscopically.
Terminal necropsy showed no macroscopic abnormalities except for unilateral hydronephrosis of 1 male in the lower dose
group.
Other findings:
- Other observations: Autopsy revealed pallor of the kidneys, liver and/or spleen in all rats that died.

Table 1: Time and number of deaths or rats dosed orally with L-carvone (Main study)

Day
Sex Dose (g/kg) Number of deaths in a group of 5 1 2 3 4 5 6 to 15
a b a b a b a b a b a b
Male 3.2 0
5 1 1
8 5 3 1 1
Female 3.2 0
5 3 2 1
8 5 2 1 2

(a): First observation

(b:) Second observation

.

Interpretation of results:
not classified
Conclusions:
The acute median lethal oral doses (LD50) and their 95% confidence limits to rats of L-carvone were estimated to be:

LD50 (combined): 5400 mg/kg bw (4600 - 6300 mg/kg bw; 95% C.I)
LD50 (male): 5900 mg/kg bw (4900 - 7100 mg/kg bw; 95% C.I)
LD50 (female): 4900 mg/kg bw (3900 - 6100 mg/kg bw; 95% C.I)
Executive summary:

In an acute oral toxicity study (86771D/ULR 138/AC), groups of fasted, young CFY (Sprague Dawley origin) (5/sex) were given single oral doses of L-carvone undiluted at 3.2, 5.0, or 8.0 g/kg bw and observed for 14 days.

LD50 (combined): 5400 mg/kg bw (4600 - 6300 mg/kg bw; 95% C.I)

LD50 (male): 5900 mg/kg bw (4900 - 7100 mg/kg bw; 95% C.I)

LD50 (female): 4900 mg/kg bw (3900 - 6100 mg/kg bw; 95% C.I)

No mortalities were observed at 3.2 g/kg bw while no rat survived administration of L-carvone of 8 g/kg bw. Clinical signs were noted at all doses and resolved towards the end of the observation period in surviving animals. Body weights were lower during first week of observation but were as anticipated at end of second week of observation. Terminal necropsy showed no treatment-related macroscopic abnormalities.

This acute oral study is classified as acceptable. It does satisfy the guideline requirement for an acute oral study (OECD 401) in the rat.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 400 mg/kg bw
Quality of whole database:
The key study was the only study available and was assigned a Klimisch score of 1. The overall quality of the database is high.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
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
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
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

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].
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) .

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)

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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The key study was the only study available and was assigned a Klimisch score of 1. The overall quality of the database is high.

Additional information

Acute oral toxicity

There is one acute oral toxicity study in rats available.

In an acute oral toxicity study (OECD 401/GLP), groups of fasted, young CFY (Sprague Dawley origin) (5/sex) were given single oral doses of L-carvone undiluted at 3.2, 5.0, or 8.0 g/kg bw and observed for 14 days. No mortalities were observed at 3.2 g/kg bw while no rat survived administration of L-carvone of 8 g/kg bw. Clinical signs were noted at all doses and resolved towards the end of the observation period in surviving animals. Body weights were lower during first week of observation but were as anticipated at end of second week of observation. Terminal necropsy showed no treatment-related macroscopic abnormalities. The LD50 (combined) was 5400 mg/kg bw (4600 - 6300 mg/kg bw; 95% C.I). The LD50 (male) was 5900 mg/kg bw (4900 - 7100 mg/kg bw; 95% C.I). The LD50 (female) was 4900 mg/kg bw (3900 - 6100 mg/kg bw; 95% C.I).

Acute dermal toxicity

There is one acute dermal toxicity study in rats available.

In an acute dermal toxicity study (OECD 402/GLP), 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. There were no treatment related clinical signs, necropsy findings, changes in body weight or signs of dermal irritation noted during the study.

Both studies are suitable to use the in human health risk assessment.

Justification for classification or non-classification

Based on the available information in the dossier, the substance L-carvone (CAS No. 6485-40-1) does not need to be classified for acute toxicity or specific target organ toxicity - single exposure when the criteria outlined in Annex I of 1272/2008/EC are applied.