[1R-(1α,2β,5α)]-2-isopropyl-5-methylcyclohexyl 5-oxo-L-prolinate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April - September 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

[1R-(1α,2β,5α)]-2-isopropyl-5-methylcyclohexyl 5-oxo-L-prolinate

EC Number:

264-935-8

EC Name:

[1R-(1α,2β,5α)]-2-isopropyl-5-methylcyclohexyl 5-oxo-L-prolinate

Cas Number:

64519-44-4

Molecular formula:

C15H25NO3

IUPAC Name:

[1R-(1α,2β,5α)]-2-isopropyl-5-methylcyclohexyl 5-oxo-L-prolinate

Test material form:

liquid

Details on test material:

Batch No: G17066-2
Substance type: Monoconstituent
CAS-No: 64519-44-4
EINECS-No: 264-935-8
Description: Yellow to pale pink liquid
Test substance storage: Keep in a tightly closed container in a cool and dry place, protected from light.
Date of manufacture: March 2017
Date of expiration: 13/03/2019

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, France
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 205 ± 9.1 g
- Fasting period before study: 1 day
- Housing: Animals were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid.
- Diet (e.g. ad libitum): Foodstuff (ENVIGO - 2016), ad libitum
- Water (e.g. ad libitum): Drinking water (tap-water from public distribution system), ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18-31°C
- Humidity: 26-70%
- Air changes: At least 10/hour
- Photoperiod: 12 hours dark / 12 hours light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

VEHICLE
- Concentration in vehicle: ca. 2000 mg/10 mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: Dimethyl Sulfoxide (DMSO) was chosen as it produced the most suitable formulation at the requested concentrations.

DOSAGE PREPARATION: In the first and second step of the study, 2.0003g and 4.0048 g of the test item were weighed and DMSO was added to two 10 mL volumetric flasks. Just before the administration, the preparations were stirred by vortex to obtain slightly colored yellow solutions. Each preparation was administered under a volume of 10 mL/kg body weight using a suitable syringe graduated fitted with an oesophageal metal canula.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 female rats

Control animals:

other: historical data

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations and mortality were recorded at 30 minutes, 1 hour, 3 hours, 4 hours, 24 hours, and then once daily for 14 days. Animals were weighed on day D0 (just before administering the test item) and then on D2, D7, and D14.
- Necropsy of survivors performed: Yes; animals were euthanized with sodium pentobarbital (Dolethal®) on D14 and macroscopic observations were noted.

Results and discussion

Preliminary study:

Not applicable

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

An absence or decrease in spontaneous activity (4/5), in Preyer’s reflex (4/5), in muscle tone (4/5) and in righting reflex (4/5) and an increase in salivation (4/5) were noted from 30 minutes or 1 hour post-dose. The animals recovered a normal behaviour at 24 hours post-dose.

Body weight:

The body weight evolution of the animals remained normal throughout the study.

Other findings:

The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of test item Questice L is higher than 2000 mg/kg body weight by oral route in the rat.
In accordance with Regulation (EC) No. 1272/2008 on classification, labelling and packaging substances and mixtures, the test item Questice L does not have to be classified. No signal word or hazard statement is required.

Executive summary:

In an acute oral toxicity study performed according to the OECD Guideline 420 and in compliance with GLP, a single dose of 2000 mg/kg bw of the test substance was given by oral gavage to a group of 5 female Sprague Dawley rats. Animals were then observed for mortality, clinical signs and body weight changes for 14 days, and were all sacrificed for macroscopic examinations.

No mortality occurred during the study. No clinical signs related to the administration of the test item were observed during the study, except for an absence or decrease in spontaneous activity (4/5), in Preyer’s reflex (4/5), in muscle tone (4/5) and in righting reflex (4/5) and an increase in salivation (4/5) were noted from 30 minutes or 1 hour post-dose. The animals recovered a normal behaviour at 24 hours post-dose. The body weight evolution of the animals remained normal throughout the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Rat Oral LD50 >2000 mg/kg bw.

Under the test conditions, the oral LD50 of the test substance is >2000 mg/kg bw in rats. In accordance with Regulation (EC) No. 1272/2008 on classification, labelling and packaging substances and mixtures, the test item does not have to be classified. No signal word or hazard statement is required.

In accordance with Globally Harmonised System of classification and labelling of chemicals (GHS), the test item Boswellia serrata extract has to be classified either Category 5 with the signal word “Warning” and the hazard statement “H303 May be Harmful if swallowed” or does not have to be classified with no signal word or hazard statement required.