Reaction mass of (E)-1-(3,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one and (E)-1-(4,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 April 2019 - 21 May 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The study is considered to be a reliability 1 as it has been conducted according to OECD Guideline for Testing of Chemicals No. 420 ‘Acute Oral Toxicity – Fixed Dose Method’ Adopted 17 December 2001 and in compliance with GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of (E)-1-(3,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one and (E)-1-(4,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one

EC Number:

950-718-1

Molecular formula:

C14H22O

IUPAC Name:

Reaction mass of (E)-1-(3,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one and (E)-1-(4,6-dimethylcyclohex-3-en-1-yl)-2-methylpent-1-en-3-one

Test material form:

liquid

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS B.V.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: 156.2 - 181.8 g
- Fasting period before study: overnight
- Housing: groups of one to five rats (of the same sex and dose group)
- Diet: 2018C Teklad Global 18% protein rodent diet (certified), ad libitum
- Water: tap water, ad libitum
- Acclimation period: At least 5 days prior to the start of dosing under test conditions after health examination

ENVIRONMENTAL CONDITIONS
temperature 22 + 2°C
relative humidity approx. 45-65 (except for deviation) (with the aim of 50 – 60%)
artificial light 6.00 a.m. - 6.00 p.m.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

Dose administration was once orally by gavage (feeding needle). The volume administered did not exceed 10 mL/kg b.w.
Based on available information on the toxicity of the test item, 300 mg/kg was chosen as the initial starting dose, since a severe toxicity which may necessitate humane euthanasia was not expected at this dose level.

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

Two animals treated as follows:
- one at 300 mg/kg
- one at 2000 mg/kg

Due to mortality and signs of systemic toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated as follows:
- four rats at 300 mg/kg
A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.

Control animals:

no

Details on study design:

Clinical observations and inspections for morbidity / mortality were performed at least three times within the first six hours after application (i.e., 30 minutes and 1 hour, 2 hours and 4 hours after dosing), thereafter at least once daily for 14 days. All surviving animals were observed for 14 days after dosing.
The nature and severity, where appropriate, of the clinical signs and the time were recorded at each observation for all individual animals. If applicable, the time of death/humane kill was recorded as precisely as possible. Observations included changes in the skin and fur, eyes and mucous membranes, and respiratory, circulatory, autonomic and central nervous system, and somatomotor activity and behavior pattern. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma.

Animals were sacrificed by carbon dioxide asphyxiation followed by cervical dislocation.
Body weights were recorded on Day 0 (prior to dosing), Day 7, and 14, or (if applicable) at death (unscheduled).
A gross necropsy was performed on all animals that died or were humanely killed during the study (if applicable) and at the end of the in-life part. Any macroscopic abnormalities were recorded. Routinely no organs or tissues were retained.

Statistics:

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.

Results and discussion

Preliminary study:

N/A

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

300 mg/kg b.w: There were no deaths during the study.

2000 mg/kg b.w: The animal was euthanized 4 hours after application due to deterioration of symptoms.

Clinical signs:

other: 300 mg/kg b.w: There were no clinical signs of reaction to treatment throughout the study. 2000 mg/kg b.w: Partially closed eyes, decreased activity, sunken flanks, piloerection, prostration, lethargy, ataxia, hunched back, and increased respiration rate

Gross pathology:

300 mg/kg b.w: One animal showed a clear liquid in the uterus. No abnormalities were noted at necropsy in the remaining animals.

2000 mg/kg b.w: Upon macrosopic examination of the animal which was sacrificed after 4 hours of dosing the stomach was found filled with diet and test item, with an aromatic, sweet smell. Furthermore hardened feces were noted proximal to the caecum.

Other findings:

Not specified

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute median lethal oral dose (LD50) to rats of PG-RAW-90-032 was demonstrated to be between 300 and 2000 mg/kg body weight.

Executive summary:

The acute median lethal oral dose (LD50) of the test substance PG-RAW-90-032, was between 300 and 2000 mg/kg body weight according to OECD Test Guideline 420 using the fixed dose procedure.