2-nitro-4-(trifluoromethyl)benzonitrile

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 Feb - 09 Mar 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

adopted 17 Dec 2001

GLP compliance:

yes

Remarks:

The test complied with the Principles of Good Laboratory Practices (GLP) of the Certification and Accreditation Administration of the People’s Republic of China (2013 revised edition).

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2-nitro-4-(trifluoromethyl)benzonitrile

EC Number:

212-298-1

EC Name:

2-nitro-4-(trifluoromethyl)benzonitrile

Cas Number:

778-94-9

Molecular formula:

C8H3F3N2O2

IUPAC Name:

2-nitro-4-(trifluoromethyl)benzonitrile

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SPF grade

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Liaoning Changsheng Biotechnology Co., Ltd., Liaoning, China
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: 201.80 - 235.18 g
- Fasting period before study: overnight before and 3 - 4 hours after exposure
- Housing: 2 animals during acclimation, individually during testing period, in hanging stainless steel cages (L32.0 cm x W28.0 cm x H20.0 cm) affixed to racks
- Diet: SPF rat/mice maintenance feed (Liaoning Changsheng Biotechnology Co., Ltd. China); ad libitum (except night before treatment and 3 - 4 h after exposure)
- Water: grade-one reverse osmosis water prepared at testing centre; ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.65 - 23.30
- Humidity (%): 41.23 - 69.80
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From 15 Feb to 09 Mar 2019

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL in the 300 mg/kg bw dose group; 200 mg/mL in the 2000 mg/kg bw dose group; actual weighed sample quantity was 0.3015 g, 0.3007 g and 1.9999 g, in the first, second, and third step, respectively.
- Justification for choice of vehicle: mixes more evenly with test material
- Lot/batch no: FN2019/01/03

MAXIMUM DOSE VOLUME APPLIED: 10 mL/Kg

DOSAGE PREPARATION:
Theoretical weighed sample quantity was calculated based on designed dose and required volume. After grinding, the test sample was weighed in an appropriate container. A suitable quantity of vehicle was added to the standard mark, magnetically stirred for at least three minutes, and labelled in preparation for use. All test sample solutions were used within 4 h. In order to ensure the uniformity of the test sample solution, prior to use, the test sample solution was stirred on a magnetic stirrer for at least five minutes, and it was stirred throughout the entire exposure process.

CALCULATION FORMULA:
test sample concentration (mg/mL) = dose (mg/kg)/exposure volume (mL/kg),
theoretical weighed sample quantity (g) = prepared volume (mL) x test sample concentration (mg/mL)/1000.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 animals per step:
300 mg/kg: 2 x 3 animals (step 1 and 2); total 6 animals
2000 mg/kg (step 3): 3 animals

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were checked twice daily, morning and night, for dead and moribund animals and time of death was recorded. The animals were weighed at introduction, at grouping, the day of exposure (fasting), at day 7 and day 14 after exposure, and at death.
- Necropsy of survivors performed: yes: gross necropsy was performed on all exposed animals (including animals that died in the test period) and gross necropsy findings were recorded in detail for each animal. Macroscopic observation comprised the tissues, organs, and contents of the animal’s thoracic cavity and abdominal cavity.
- Clinical signs including body weight: Detailed observation was performed once within 30 minutes on the day of exposure and at one, two, and four hours. Subsequently, symptoms were observed once daily and observation continued for 14 days. All observation results were fully recorded and individual records were kept for each animal. Changes to animal fur, eyes, mucous membranes, respiratory system, circulatory system, and nervous system were observed and recorded in detail, with particular attention to changes in limb movements and behaviour.

Statistics:

not performed

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off: 500 mg/kg

Mortality:

300 mg/kg bw: No mortality occurred during the test period.
2000 mg/kg bw: 3/3 females died: one female died on the day of exposure, the remaining two females died one day after exposure.

Clinical signs:

lethargy (hypoactivity)

observations of tremors

other:

Body weight:

other body weight observations

Remarks:

Surviving animals gained weighed throughout the study.

Gross pathology:

300 mg/kg bw: No abnormalities were seen in gross necropsy of any of the animals.
2000 mg/kg bw: One animal exhibited abdominal distention. One animal had brown pigment changes in the lungs and the gastric contents were light yellow. In the third animal, the gastric contents were also light yellow.

Any other information on results incl. tables

Table 1: Acute oral toxicity

Dose [mg/kg bw]	Mortality	Clinical signs
	N*	N*
Females
300	0/6	0/6
2000	3/3	1/3

*Number of animals/ number of animals used

Applicant's summary and conclusion

Interpretation of results:

other: Acute Oral 4 (H302) according to Regulation (EC) No 1272/2008

Conclusions:

In this acute oral toxicity study in rats a LD50 value of > 300 - 2000 mg/kg bw was determined. The LD50 cut-off was 500 mg/kg bw. The substance thus meets the criteria for Acute Oral 4 (H302) according to Regulation (EC) No 1272/2008.