Cymbopogon winterianus, ext.

Endpoint summary

Description of key information

Acute oral toxicity: 300 >LD50< 2000 mg/kg bw (OECD 420)

Acute dermal toxicity: LD50> 2000 mg/kg bw (OECD 402)

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:

27 August 2015 to 22 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, performed in accordance with OECD Guideline 420

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

GLP compliance:

yes (incl. QA statement)

Remarks:

statement of compliance presented

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Age at study initiation: 8 to 12 weeks
- Fasting period before study: overnight fast before dosing and for 3-4 hours after dosing
- Housing: in groups, up to four animals, suspended solid-floor polypropylene cages furnished with woodflakes
- Diet: ad libitum, 2014C Teklad Global Rodent diet
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30 to 70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12

Route of administration:

oral: gavage

Vehicle:

arachis oil

Remarks:

BP; only at 300 mg/kg bw

Details on oral exposure:

The veicle was used only with the 300 mg/kg bw dose. At the high dose level of 2000 mg/kg bw the test item was supplied as such.
- Concentration in vehicle: 30 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: the test substance did not dissolve or suspend in water
- Rationale for the selection of the starting dose: no toxicity data available and hence, 300 mg/kg bw/d was chosen as a starting point.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

5 animals at 300 and 1 animal at 2000 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of clinical observations : 30 min, 1, 2, & 4 h after dosing and then daily for up to 14 days
- Body weight recording: at Day 0 (after dosing), Day 7 and 14
- Morbidity/mortality: twice daily
- Necropsy of survivors performed: yes, external examination and opening of the abdominal and thoracic cavities
- Other examinations performed: no

Statistics:

not applicable

Preliminary study:

Animal tested with 2000 mg/kg bw: hunched posture, increased respiratory rate, labored respiration, ataxia, lethargy, pallor of the extremities, hypothermia and pilo-erection. The animal was humanely killed due to signs of severe enduring.
Gross necropsy: Patchy pallor of the liver and epithelial sloughing of the gastric mucosa

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality seen at the dose level of 300 mg/kg bw.

Clinical signs:

other: No signs of toxicity were observed.

Gross pathology:

No adverse effects detected.

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 of Citronella oil (Cymbopogon winterianus, ext.) was determined to be between 300 and 2000 mg/kg bw, under the conditions of this test. The test substance is considered as harmful if swallowed (CLP criteria, Acute Tox. 4, H302).

Executive summary:

In an acute oral fixed dose toxicity study, performed according to OECD 420, female Wistar rats were administered Citronella oil (Cymbopogon winterianus, ext.) by gavage. In this stepwise approach, an initial test was performed with the dose levels of 300 mg/kg bw in arachis oil and 2000 mg/kg bw as such (no vehicle) administered to single animals. The results lead to an additional group of four animals treated with a single dose of 300 mg/kg bwin arachis oil, followed by a 14-day observation period. Mortality, clinical signs of toxicity and body weights were recorded. Necropsy was performed in all animals.

 

Treatment with 2000 mg/kg bw induced toxicity, such as hunched posture, increased respiratory rate, labored respiration, ataxia, lethargy, pallor of the extremities, hypothermia and pilo-erection;the animal was humanely killed. No deaths or clinical signs of toxicity were seen at the dose level of 300 mg/kg bw. Body weights appeared unaffected by the treatment. Patchy pallor of the liver and epithelial sloughing of the gastric mucosa were detected at necropsy of the animal treated with 2000 mg/kg bw, while no abnormalities were seen in any of the animals treated with the lower dose. The oral LD50 was considered to be between 300 and 2000 mg/kg bw.The test item needs be classified according to the CLP 1272/2008/EC criteria, as harmful if swallowed, Acute Tox.4, H302.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

The only study available, is a recent GLP, Guidance study and it is considered appropriate to be used as the basis for the chemical safety assessment.The LD50 was determined to be between the range of 300 and 2000 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:

09 September 2015 to 30 September 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, performed according to the OECD Guideline 402.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

Initially, 1 animal/sex was given a single dose of 2000 mg/kg body weight. Based on the result, an additionalgroup of 8 animals was treated accordingly. This deviation does not compromise the result of the study.

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Oxon, UK
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: at least 200 g
- Fasting period before study: overnight fast before dosing and for 3-4 hours after dosing
- Housing: indivudually during dosing and till 24 h post-treatment, in groups up to 4 animals of same sex rest of the experiment, suspended solid-floor polypropylene cages furnished with woodflakes
- Diet: ad libitum, 2014C Teklad Global Rodent diet
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30 to 70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure, % coverage: 10% of the total body surface area
- Type of wrap if used: semi-occlusive self-adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): cotton wool moistened with arachis oil BP
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.26 mL/kg
- Constant volume or concentration used: yes

VEHICLE no vehicle used

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

Initially, 2 animals (1 male, 1 female) were given a single dermal application of the test item at a dose level of 2000 mg/kg body weight. Based on the result, an additionalgroup of 8 animals was treated accordingly.
- Duration of observation period following administration: 14 days
- Frequency of clinical observations : 30 min, 1, 2, & 4 h after dosing and then daily for up to 14 days
- Body weight recording: at Day 0 (after dosing), Day 7 and 14
- Morbidity/mortality: twice daily
- Necropsy performed: yes, external examination and opening of the abdominal and thoracic cavities
- Other examinations performed: yes, irritation scoring assigned once per day for the whole observation period

Statistics:

Not applicable

Preliminary study:

No signs of systemic toxicity observed at any of the two animals treated during the initial test.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was detected

Clinical signs:

other: No signs of toxicity observed at any the two animals treated

Gross pathology:

No abnormalities seen

Other findings:

- Other observations: no signs of dermal irritation

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The dermal LD50 of Citronella oil (Cymbopogon winterianus, ext.) was estimated to be higher than 2000 mg/kg bw, under the conditions of this test and hence, the test substance is practically non toxic.

Executive summary:

In an acute dermal toxicity study, performed according to OECD 402, male and female Wistar rats were administered with a single dose of unchanged Citronella oil (Cymbopogon winterianus, ext.), for 24 h kept in place with a semi- occlusive dressing. An initial test was performed with one male and one female animal. The results lead to an additional group of four animals/sex treated with a single same dose of 2000 mg/kg bw, followed by a 14-day observation period. Mortality, clinical signs of toxicity and body weights were recorded. Irritation scorings were given. Necropsy was performed in all animals.

Treatment with the test item did not result in any deaths, clinical signs of toxicity or macroscopical abnormalitites. Transient body weight losses were recorded in two animals, but were deemed acceptable variations for the species tested. The dermal LD50 of the test substance was established to be higher than 2000 mg/kg bw, under the conditions of this test. The test shall not be classified for acute oral toxicity, as set down by the CLP 1272/2008/EC criteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

One acute oral toxicity test is available for Citronella oil (C. winterianus ext.), in which the fixed dose procedure was applied Single doses: 300 and 2000 mg/kg bw were given by gavage to female Wistar rats. The LD50 was estimated to be in the range of 300 and 2000 mg/kg bw, due to severe toxicity and deliberate death at the highest administered dose.

Acute dermal toxicity was tested in a standard acute limit test, during which Citronella oil (C.winterianus ext.) was applied dermally for 24 h to the skin of 5 male and 5 female Wistar rats, at a dose of 2000 mg/kg bw. No mortality was noted, and hence, the dermal LD50 is considered higher than 2000 mg/kg bw.

Justification for classification or non-classification

The oral LD50 of Citronella oil (Cymbopogon winterianus, ext.) was found to be between 300 and 2000 mg/kg bw. Under the conditions of this test, the substance should be classified for acute oral toxicity (H302 / Acute Tox. 4) in accordance with the criteria outlined in Annex I of CLP (1272/2008/EC).

The dermal LD50 was estimated to be higher than 2000 mg/kg bw. Under the conditions of this test, Citronella oil (Cymbopogon winterianus, ext.) does not need to be classified for acute dermal toxicity in accordance with the criteria outlined in Annex I of CLP (1272/2008/EC).