Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
GLP compliance:
yes (incl. certificate)
Remarks:
statement of compliance presented
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Citronella oil (Cymbopogon winterianus, ext.)
- Physical state: liquid (pale yellow)
- Analytical purity: 100%
-Storage conditions: approximately 4 °C in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals 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

Administration / exposure

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

Results and discussion

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
Effect levels
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:
No signs of toxicity were observed.
Body weight:
The treatment did not have any effect on the body weights.
Gross pathology:
No adverse effects detected.

Applicant's summary and conclusion

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.