Litsea cubeba, ext.

Endpoint summary

Description of key information

Acute oral toxicity: LD50 > 5000 mg/kg bw (similar to OECD 401)

Acute dermal toxicity: LD50 = 4800 mg/kg bw (similar to OECD 402)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

Test was conducted according to methods similar to OECD guideline 401 (limit test) and was performed pre-GLP. A concise description of the protocol is available and results are reported clearly.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Remarks:

pre-GLP

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: approximately 200 g
- Fasting period before study: 16-18 hrs
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum

Route of administration:

oral: gavage

Vehicle:

not specified

Details on oral exposure:

No data available

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

10 males

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: Daily
- Necropsy of survivors performed: yes
- Other examinations performed: symptomatology

Preliminary study:

Not relevant

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

Mortality was observed in one rat.

Clinical signs:

other: No effects observed

Gross pathology:

Necropsy performed: lungs red throughout in one rat

Other findings:

Not reported

Interpretation of results:

other: not classified

Remarks:

according to the classification criteria outlined in Annex I of 1272/2008/EC (CLP)

Conclusions:

The oral LD50 value of Litsea Cubeba Oil in rats was established as exceeding 5000 mg/kg body weight, under the conditions of this study. The substance therefore does not need to be classified according to the classification criteria outlined in Annex VI of 67/548/EEC (DSD) and Annex I of 1272/2008/EC (CLP).

Executive summary:

A single 5000 mg/kg bw dose of Litsea Cubeba oil was administered by oral gavage to 10 male Wistar rats. The animals were observed for 14 days. No symptoms were noted. Mortality was observed in one rat. The oral LD50 value of Litsea Cubeba oil in rats was established as exceeding 5000 mg/kg body weight, under the conditions of this study. The substance therefore does not have to be classified according to the classification criteria outlined in Annex VI of 67/548/EEC (DSD) and Annex I of 1272/2008/EC (CLP).

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

3 November 1976 - 29 December 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test was conducted according to methods similar to OECD guideline 401 and was performed pre-GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

(study was used for range-finding)

GLP compliance:

no

Remarks:

performed pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

mouse

Strain:

not specified

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Doses:

1760, 4400 and 8800 mg/kg bw

No. of animals per sex per dose:

- 1760 mg/kg bw:: 1 animal per sex
- 4400 mg/kg bw:: 3 animals per sex
- 8800 mg/kg bw:: 1 animal per sex

Control animals:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

4 400 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 3 out of 6 animals died (all female)

Clinical signs: Animals of the two highest dose groups were hypothermic, somnolent or comatose, showing signs of stress and exhibiting laboured breathing within 30 minutes after treatment. Mice of the low dose group appeared unaffected by treatment.

Mortality: Mice of the highest dose group died within 2 and 18 hours after treatment. Three female mice dosed at 4400 mg/kg bw were dead within 18 hours after treatment, but the male mice at this dose level recovered within 42 hours. No mortality was observed in the lowest dose group.

Interpretation of results:

other: not classified

Remarks:

according to the classification criteria outlined in Annex I of 1272/2008/EC (CLP)

Conclusions:

Under the conditions of this study, mice exposed to Litsea cubeba by oral gavage showed acute effects and mortality at the 4400 and 8800 mg/kg bw dose. Based on these results an LD50 of 4400 mg/kg bw was established. The substance therefore does not need to be classified according to the classification criteria outlined in Annex I of 1272/2008/EC (CLP).

Executive summary:

An acute oral toxicity according to similar methods as described in OECD guideline 401 was performed with test substance Litsea cubeba essential oil. Three doses (2, 5, 10 ml/kg bw) were administered by oral gavage to male and female mice (1, 3 and 1 animal(s)/sex, respectively). Clinical signs (incl. mortality) were noted and postmortem examinations were performed.

Animals of the two highest dose groups were hypothermic, somnolent or comatose, showing signs of stress and exhibiting laboured breathing within 30 minutes after treatment. Mice of the low dose group appeared unaffected by treatment. Mice of the highest dose group died within 2 and 18 hours after treatment. Three female mice dosed at 4400 mg/kg bw were dead within 18 hours, but the male mice at this dose level recovered within 42 hours. No mortality was observed in the lowest dose group.

Under the conditions of this study, mice exposed to Litsea cubeba essential oil by oral gavage showed acute effects and mortality at the 4400 and 8800 mg/kg bw dose. Based on these results an LD50 of 4400 mg/kg bw was established. The substance therefore does not need to be classified according to the classification criteria outlined in Annex I of 1272/2008/EC (CLP).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Quality of whole database:

The key study was conducted in rats according to methods similar to OECD guideline 401 (limit test) and was performed pre-GLP. A concise description of the protocol is available and results are reported clearly. An adverse effect (mortality in 1 out of rats) was observed. However based on this result, no discriminating dose could be derived LD50 > 5000 mg/kg bw.

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:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Test was conducted according to methods similar to OECD guideline 402 and was performed pre-GLP. A concise description of the protocol is available and results are reported clearly.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Remarks:

pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: Approximately 2 kg
- Housing: 2 animals per cage
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum

ENVIRONMENTAL CONDITIONS
-Temperature controlled room

Type of coverage:

occlusive

Vehicle:

not specified

Details on dermal exposure:

TEST SITE
- Area of exposure: Abdominal area (skin was abraded)
- Type of wrap if used: 2 single layers of gauze and impervious material

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 24 hrs

Duration of exposure:

24 hrs

Doses:

1250, 2500, 5000 mg/kg bw

No. of animals per sex per dose:

Four animals per dose, with a total of 12 animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: daily
- Necropsy of survivors performed: yes
- Other examinations performed: symptomatology, skin irritation

Statistics:

Not relevant

Preliminary study:

Not relevant

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

4 800 mg/kg bw

Based on:

test mat.

95% CL:

2 400 - 9 600

Mortality:

Mortality was observed in two out of four animals in the highest dose (5000 mg/kg bw) on the third and sixth day, and in one out of four animals on the fourth day in the 2500 mg/kg dose group.

Clinical signs:

other: For the two lowest dose groups no clinical signs were noted, while in the highest doser group anorexia, ptosis and ataxia were noted in 2 animals (not specified whether these were the same animals). In all rabbits and at all doses marked redness and moder

Gross pathology:

Necropsy:
- 2500 mg/kg bw: evidence of extreme diarrhea, extreme dermal and subdermal irritation in 1 animal
- 5000 mg/kg bw: liver blotchy in 1 animal, subdermal irritation in 1 animal, no visible lesion in 1 animal

Other findings:

Not reported

Interpretation of results:

other: Not classified

Remarks:

according to the classification criteria outlined in Annex I of 1272/2008/EC (CLP)

Conclusions:

Under the conditions of this study, dermal application of Litsea Cubeba oil induced mortality at a dose of 2500 mg/kg bw (1/4 animals) and at a dose of 5000 mg/kg bw (2/4 animals). The LD50 is established at 4800 mg/kg bw (CI: 2400-9600) and therefore the substance does not need to be classified for acute dermal toxicity according to the criteria outlined in Annex I of 1272/2008/EC (CLP).

Executive summary:

Three dose levels of Litsea Cubeba oil (1250, 2500, 5000 mg/kg bw) were applied dermally to the skin of 12 rabbits (4 per group). The rabbits were observed for 14 days thereafter for mortality and symptomatology (skin irritation).

No mortality was observed in the lowest dose group. In the 2500 mg/kg bw group, mortality was observed in 1 out of 4 animals. In the 5000 mg/kg bw group 2 out of 4 animals died. At the highest dose level, anorexia was observed in 2 animals, ptosis in 2 animals and ataxia in 2 animals. At 2500 mg/kg bw evidence of extreme diarrhea, extreme dermal and subdermal irritation in 1 animal were observed. At 5000 mg/kg bw a blotchy liver was observed in 1 animal, subdermal irritation in 1 animal and no visible lesions were observed in 1 animal.

Based on the conditions of this study, the LD50 was established to be 4800 mg/kg bw (confidence interval of 2400-9600 mg/kg bw) and therefore the substance does not need to be classified for acute dermal toxicity according to the criteria outlined in Annex I of 1272/2008/EC (CLP).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

4 800 mg/kg bw

Quality of whole database:

Test was conducted according to methods similar to OECD guideline 402 and was performed pre-GLP. A concise description of the protocol is available and results are reported clearly.

Additional information

Acute oral toxicity

In the key study a single 5000 mg/kg bw dose of Litsea Cubeba oil was administered by oral gavage to 10 male Wistar rats. No symptoms were noted. Mortality was observed in one rat. The oral LD50 value of Litsea Cubeba oil in rats was established as exceeding 5000 mg/kg bw. This result is supported by another study (similar to OECD 401, performed in mice) in which an LD50 of 4400 mg/kg bw was established.

Acute dermal toxicity

In the key study three dose levels of Litsea Cubeba oil (1250, 2500, 5000 mg/kg bw) were applied dermally to the skin of 12 rabbits (4 per group). Mortality was observed in 1 out of 4 animals at a dose of 2500 mg/kg bw and in 2 out of 4 animals at a dose of 5000 mg/kg bw. No mortality occurred in the lowest dose group. Based on these results, the LD50 was established to be 4800 mg/kg bw (confidence interval of 2400-9600 mg/kg bw).

Justification for selection of acute toxicity – oral endpoint

The selected study is the key study for this endpoint.

Justification for selection of acute toxicity – dermal endpoint

The selected study is the key study for this endpoint.

Justification for classification or non-classification

Based on the available information, Litsea cubeba oil has shown to be non-toxic after oral exposure. Therefore, the substance does not need to be classified for Acute Oral Toxicity in accordance with the criteria outlined in Annex I of 1272/2008/EC (CLP/EU-GHS).

Based on the available information, Lisea cubeba oil has shown to be non-toxic in contact with skin. Therefore, the substance does not need to be classified for Acute Dermal Toxicity in accordance with the criteria outlined in Annex I of 1272/2008/EC (CLP/EU-GHS).