Reaction mass of methyl 2-[[(E)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate and methyl 2-[[(Z)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5 April 2016 - 25 April 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

An assessment of acute toxicity is required to fulfil REACH Annex VII information requirements. The acute oral toxicity study was conducted according to OECD TG 423, GLP and is considered reliable without restriction (Klimisch 1).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 2-[[(Z)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate

Molecular formula:

C17H21NO2

IUPAC Name:

Methyl 2-[[(Z)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate

Constituent 2

Reference substance name:

Methyl 2-[[(E)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate

Molecular formula:

C17H21NO2

IUPAC Name:

Methyl 2-[[(E)-(2,4-dimethylcyclohex-3-en-1-ylidene)methyl]amino]benzoate

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Species, strain: Sprague-Dawley (Crl:CD(SD)), SPF
- Age at study initiation: 8 weeks
- Weight at study initiation: 179.9-190.2 g
- Fasting period before study: Approximately 16 hours
- Housing: Housed individually in stainless wire mesh cages (260 x 350 x 210 mm)
- Diet (e.g. ad libitum): Pelleted rodent chow, ad libitum
- Water (e.g. ad libitum): Public tap water in Cheongju-si was filtered and irradiated by ultraviolet light and provided ad libitum.
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1−23.2°C
- Humidity (%): 45.1−54.2%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle, 150-300 Lux

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 500 mg/mL
- Lot/batch no. (if required): MKBV2080V

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION (if unusual): The test item was weighed into a bottle and a small amount of corn oil was added. The test item was dissolved using a vortex mixer. Corn oil was gradually added to yield the desired concentration.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Due to the low expected toxicity of the test substance, based on information supplied by the sponsor, a starting dose of 5,000 mg/kg was administered in one animal for this study (Step 1).

Doses:

5000 mg/kg bodyweight

No. of animals per sex per dose:

1 for Step 1, 2 for Step 2

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for mortality, general condition and clinical signs (type, severity, time of onset and recovery) at 30 minutes after dosing and at 1, 2, 4 and 6 hours after dosing on Day 0 and once daily thereafter for 14 days (Day 1−Day 14). The body weight was recorded prior to dosing on Day 0 and on Days 1, 3, 7 and on the day of necropsy, Day 14.
- Necropsy of survivors performed: Yes
- Other examinations performed: clinical signs and body weight

Statistics:

Statistical analysis was not performed. Mean scores and values are determined.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

All animals at 5,000 mg/kg survived the duration of the study. There were no effects on mortality.

Clinical signs:

other: Chromaturia (abnormal colour of the urine) were observed in all animals at 6 hours after dosing at 5,000 mg/kg. Chromaturia, compound-coloured stool, decrease in locomotor activity, decrease of faecal volume, mucous stool, soiled perineal region and/or ab

Gross pathology:

No abnormal morphological findings were observed in any animal at 5,000 mg/kg.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In an acute oral toxicity study conducted according to OECD TG 423, GLP and considered reliable without restriction (Klimisch 1), the LD50 (cut-off) was determined to be ≥ 5,000 mg/kg bw.

Executive summary:

The acute oral toxicity of the test item was determined by the Acute Toxic Class Method in female Sprague-Dawley rats. Three animals were administered a single oral dose of 5000 mg/kg body weight (bw) by intubation. Mortality, clinical signs of toxicity and body weight were observed for 14 days following administration. Mortality was not observed at any tested dose level, and the LD50 was expected to be >5000 mg/kg bw. Effects considered to be test substance related included chromaturia, compound-colored stool, decrease in locomotor activity, decrease of fecal volume, mucous stool, soiled perineal region, abnormal gait, suppression of body weight gain and/or decreased bodyweights on Days 1 and 2.

 

This study is considered to be reliable without restrictions (Klimisch 1) as it was GLP-compliant and was performed according to OECD guideline 423. There is no evidence of a relevant intrinsic acute oral toxicity requiring classification or substance specific risk mitigation measures (RMM).