Essential oil of Citrus sinensis (Rutaceae) peel, terpene-free

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 09 June 2020 to 25 June 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 423 without deviations being an impact on the conclusion of the study (See "Principles of method if other than guideline").

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

17 December 2001.

Deviations:

yes

Remarks:

See below "Principles of method if other than guideline".

Principles of method if other than guideline:

Deviations to the study plan:
- a relative humidity higher than 70% was registered from 23 to 25 June 2020. The maximum value measured was 77%.
- A temperature higher than 25°C was registered on 21 and 23 June 2020. The maximum value measured was 28%. As no effect was noted on the health of the animals, this deviation is considered as without impact on the conclusion of the study.

GLP compliance:

yes (incl. QA statement)

Remarks:

Signed on 2019-10-14

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Essential oil of Citrus sinensis (Rutaceae) peel, terpene-free

EC Number:

951-680-9

IUPAC Name:

Essential oil of Citrus sinensis (Rutaceae) peel, terpene-free

Test material form:

liquid

Details on test material:

Batch No. 0000448007
CAS No. not applicable
EC No. 951-680-9
Form: liquid
Colour: orange-yellow to green-yellow
Date received: 25 May 2020
Production date: 13 January 2020
Expiry date: 12 January 2021
Purity: 100% UVCB
Storage: fridge (6°C+/-3°C), darkness

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle, France)
- Age at study initiation: 8 weeks old
- Weight at study initiation: 180 - 203 g
- Housing: housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid.
- Diet: foodstuff (ENVIGO, 2016), ad libitum but food was removed at D-1 and then redistributed 4 hours after the test item administration.
- Water: tap water from public distribution system, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70%
- Air changes: at least ten changes per hour.
- Photoperiod: 12 h light/12 h darkness.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.30 mL/kg bw (corresponding to 2 g/kg bw)

ADMINISTRATION OF TEST ITEM:
Animals received an effective dose of 2000 mg/kg bw of the test item used as supplied by the Sponsor, administered by by gavage under a volume of 2.30 mL/kg bw using a suitable syringe graduated fitted with an oesophageal metal canula.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed 30 min, 1, 3 and 4 hours after test item administration and thereafter once daily for 14 days. Animals were weighed pretest (Day 0) and on Day 2, 7 and 14.
- Necropsy of survivors performed: Yes; Animals were killed on Day 14 with sodium pentobarbital (Dolethal) euthanasia and subjected to macroscopic examination.

Statistics:

None

Results and discussion

Preliminary study:

Not applicable

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality occurred during the study.

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs related to the administration of the test item were observed during the study.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal treatment related changes. Consequently, no microscopic examination of organs was performed.

Other findings:

None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the oral LD50 for test substance is higher than 2000 mg/kg bw and the LD50 cut-off may be considered to be higher than 5000 mg/kg bw in female rats. Therefore, the substance is not classified according to the annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an acute oral toxicity study (limit test) performed according to OECD Guideline 423 and in compliance with GLP, 6 female Sprague Dawley rats were given a single oral (gavage) dose of test item at 2000 mg/kg bw. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and were all sacrificed for macroscopic examination.

No mortality occurred during the study. No clinical signs related to the administration of the test item were observed during the study. The body weight evolution of the animals remained normal throughout the study, similar between treated and control animals. The macroscopical examination of the animals at the end of the study did not reveal treatment-related changes.

 

Under the test conditions, the oral LD50 for test substance is higher than 2000 mg/kg bw and the LD50 cut-off may be considered to be higher than 5000 mg/kg bw in female rats. Therefore, the substance is not classified according to the annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for acute oral toxicity endpoint.