T-A06

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 September 2020 to 16 September 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

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:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

ethyl (2E)-3-(cyclopropylamino)-2-[(2,4-dichloro-3-methylphenyl)carbonyl]prop-2-enoate

Molecular formula:

C16H17Cl2N1O3

IUPAC Name:

ethyl (2E)-3-(cyclopropylamino)-2-[(2,4-dichloro-3-methylphenyl)carbonyl]prop-2-enoate

Test material form:

solid: particulate/powder

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)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8weeks
- Weight at study initiation: The animals were weighed on day D0 (just before administering the test item) then on D2, D7, and on D14.
- Fasting period before study: Day 1, food available 4 hours after test item administration
- Housing: group of three in solid-bottomed clear polycarbonate cages with a
stainless-steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air-conditioned animal husbandry.
- Diet (e.g. ad libitum): foodstuff (ENVIGO - 2016) were supplied ad libitum. Food was removed on D-1 and then redistributed 4 hours after the test item administration
- Water (e.g. ad libitum): Drinking water (tap-water from public distribution system) was supplied ad libitum
- Acclimation period: at least 5 days

- Method of randomisation in assigning animals to test and control groups

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25
- Humidity (%): 30 - 70
- Air changes (per hr): 10x/hr
- Photoperiod (hrs dark / hrs light): 12hrs dark / 12 hrs light

IN-LIFE DATES: From: 01 September 2020 To: 16 September 2020

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 1.0057g/5ml and 2.0053/10ml
- Amount of vehicle (if gavage): 10ml/kg
- Justification for choice of vehicle: it produced the most suitable formulation at the requested concentration


MAXIMUM DOSE VOLUME APPLIED: 2000mg/kg, 10ml/kg


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: As no excessive systemic toxicity of the test item was anticipated at the dose of 2000 mg/kg body weight, the first test dose was 2000 mg/kg body weight

Doses:

2000mg/kg bw

No. of animals per sex per dose:

Preliminary:1 female
Main: 4 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (or other?) 14 days
- Frequency of observations and weighing: The animals were weighed on day D0 (just before administering the test item) then on D2, D7, and on D14. Systematic examinations were carried out to identify any behavioural or toxic effects on the major physiological functions at 30 minutes, 1 hour, 3 hours, 4 hours, on day 1 and day 2 and during 14 days following the administration of the test item
- Necropsy of survivors performed: yes
- Clinical signs including body weight : were recorded
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Spontaneous activity, Preyer’s reflex (noise), Respiratory rate, Convulsions, Tremors, Body temperature, Muscle tone, Palpebral opening, Pupil appearance, Salivation, Lachrymation, Righting reflex and Back hair appearance, MORTALITY, BODY WEIGHT EVOLUTION, Organ observations

Statistics:

Not performed

Results and discussion

Preliminary study:

Preliminary Study at 2000mg/kg bw did not produce any signs of toxicity in a single female rat. According to this result, 4 more female rats recieved the same dose of 2000mg/kg bw. 

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

None

Body weight:

No abnormalities

Gross pathology:

Nothing to report

Any other information on results incl. tables

Body Weight and gains in grams

FEMALES	D0		D2	D2-D0	D7	D7-D0		D14	D14-D0
Rf 5165	187		201	14	222	35		247	60
Rf 5166 Rf 5167 Rf 5168 Rf 5169	182 205 210 208		190 210 222 231	8 5 12 23	215 233 250 247	33 28 40 39		232 251 265 269	50 46 55 61
MEAN Standard deviation	198.4 12.9		210.8 16.3	12.4 6.9	233.4 15.2	35.0 4.8		252.8 14.8	54.4 6.4

Applicant's summary and conclusion

Interpretation of results:

other: Not classified (CLP Regulation EC no. 1272/2008).

Conclusions:

The LD50 of the test item is higher than 2000 mg/ kg body weight by oral route in the rat.

Executive summary:

The oral acute toxicity of the test item was determined according to OECD Guideline 420 under GLP conditions. As no excessive systemic toxicity of the test item was anticipated at the dose of 2000 mg/kg body weight, the first test dose was 2000 mg/kg body weight. A single female 8 week old non-pregnang and nulliparous Sprague Dawley rat recieved 2000mg/kg bw via oral gavage in DMSO. No signs of toxicity were noted and 4 further female rats recieved the same dose of 2000mg/kg. No mortality occured during the study. No clinical signs or abnormal changes in body weight were recorded. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes. Thus, the LD50 of the test item is higher than 2000 mg/ kg body weight by oral route in the rat.