N-(2-chloroethyl)-N-ethylaniline

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-12-22 - 2017-02-15 (Experimental)

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

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147

Version / remarks:

amended 10 December 2002

Deviations:

no

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Health and Welfare

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N-(2-chloroethyl)-N-ethylaniline

EC Number:

245-082-0

EC Name:

N-(2-chloroethyl)-N-ethylaniline

Cas Number:

22564-43-8

Molecular formula:

C11H16ClN

IUPAC Name:

N-(2-chloroethyl)-N-ethylaniline

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 (UK) Limited, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 156 - 168 g
- Fasting period before study: over night
- Housing: The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet & Water (e.g. ad libitum): With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 °C
- Humidity (%): 30-70%
- Air changes (per hr): 15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

arachis oil

Remarks:

for 300 mg/kg bw dose only; for 2000 mg/kg dose level the test item was used as supplied.

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL
- Justification for choice of vehicle: Arachis oil BP was used because the test item did not dissolve/suspend in distilled water.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.

Doses:

2000 mg/kg bw / 300 mg/kg bw

No. of animals per sex per dose:

1 (2000 mg/kg), 5 (300 mg/kg)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.
Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Results and discussion

Preliminary study:

Dose Level - 2000 mg/kg:
- Mortality: The animal was found dead 1 day after dosing.
- Clinical Observations: No signs of systemic toxicity were noted during the observation period.
- Body Weight: Body weight loss was noted during the observation period.
Necropsy: Abnormalities noted at necropsy were dark liver and dark kidneys and hemorrhage of the gastric mucosa and non-glandular epithelium of the stomach.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

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

Based on:

test mat.

Mortality:

Dose Level - 300 mg/kg: There were no premature deaths.

Clinical signs:

Dose Level - 300 mg/kg: Hunched posture and tiptoe gait were noted in all animals. Pilo-erection was also noted in the initial treated animal. Animals appeared normal 3 or 5 days after dosing.

Body weight:

Dose Level - 300 mg/kg: All animals showed expected gains in body weight over the observation period.

Gross pathology:

Dose Level - 300 mg/kg: Thickened non-glandular epithelium of the stomach was noted at necropsy of two animals. No abnormalities were noted at necropsy of three animals.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The study was conducted under GLP according to OECD guideline 420 and EU method B1 bis on the registered substance itself. The method is to be considered scientifically reasonable without any deviations affecting the integrity or validity of the study. The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight.

Executive summary:

The study according to OECD 420 and EU method B1 bis (GLP) was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a solution in arachis oil BP, at a dose level of 300 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

The animal treated at a dose level of 2000 mg/kg was found dead 1 day after dosing. There were no deaths at a dose level of 300 mg/kg.

Signs of systemic toxicity noted at a dose level of 300 mg/kg were hunched posture, pilo-erection and tiptoe gait. Animals treated at a dose level of 300 mg/kg appeared normal 3 or 5 days after dosing.

Surviving animals showed expected gains in body weight.

Abnormalities noted at necropsy of the animal treated at a dose level of 2000 mg/kg were dark liver and dark kidneys and hemorrhage of the gastric mucosa and non-glandular epithelium of the stomach. Thickened non-glandular epithelium of the stomach was noted at necropsy of two animals treated at a dose level of 300 mg/kg. No abnormalities were noted at necropsy of three animals treated at a dose level of 300 mg/kg.

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight (Globally Harmonized Classification System - Category 4).

The test item was also classified as Category 4 according to Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures. It is reasonable to assume that the Signal Word "Warning" and the Hazard Statement "H302: Harmful if swallowed" are therefore required.