methyl (2Z)-2-(2-nitrobenzylidene)-3-oxobutanoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed in accordance with OECD Gideline No. 423.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

methyl (2Z)-2-(2-nitrobenzylidene)-3-oxobutanoate

EC Number:

810-021-4

Cas Number:

111304-31-5

Molecular formula:

C12H11NO5

IUPAC Name:

methyl (2Z)-2-(2-nitrobenzylidene)-3-oxobutanoate

Test material form:

solid: crystalline

Details on test material:

Name: Niliden (Nifedipine Intermediate 11)
CAS: 111304-31-5
Batch No.: ND-78
Description: A white to pale yellow crystalline substance
Date of analysis: 10 2014
Expiry date: 10 2015
Storage: in the refrigerator (2-8 °C)

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Experimental animals

Species and strain: Crl: (WI)BR rats
Source: Toxi Coop ZRT. Cserkesz u. 90. 103 Budapest, Hungary
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The wistar rats as a rodent is one of the standard species of acute toxicity studies
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 9 weeks old in first and second step
Body weight range
at starting (first step): 212-215 g
Body weight range
at starting (first step): 207-209 g
Acclimatization time: 12 days in first step, 13 days in second step

Husbandry

Animal health: Only healthy animals were used for the study.
Health status was certified by the study director.
Room: 5/I (E building)
Housing: Group caging (3 animals/cage)
Cage type: Type II polypropylene/polycarbonate.
Bedding: Laboratory bedding
Light: Artificial light, from 6 a.m. to 6 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 10-15 air exchanges/hour by central air condition system

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Methyl cellulose solution (1 %)

Details on oral exposure:

The test item with a suitable chemical purity, analytical certificate of the product was supplied by the Sponsor.
All doses were formulated in the vehicle. Concentration of formulations was adjusted to maintain a treatment volume of 10 mL/kg bw.
The test item was applied in a concentration of 200 mg/mL. Formulations were prepared just before the administration and were stirred continuously during the treatment.

Doses:

Starting dose was selected on the basis of the available information of the test item.
The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats.

No. of animals per sex per dose:

Three female rats in the first step at 2000 mg/kg bw dose level, further three female rats in the second step at 2000 mg/kg bw dose level again.

Control animals:

no

Details on study design:

-Dosages: Starting dose was selected on the basis of the available information of the test item.
-Procedure: A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The day before treatment the animals were fasted. The food but not water was witheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.
-Duration of the experimental period: 12 days in first step and 13 days in second step of acclimatization, treatment's day, 14 days post-treatment observation period including the treatment day, necropsy on Day 15.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

approximate LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality

Clinical signs:

In group 1 treated with 2000 mg/kg dose clinical sign of reaction comprised of decreased activity (4 cases out of 57 observations), abnormal gait (4/57), incoordination (3/57), blood around the eyes (2/57), decreased righting reflex (5/57), decreased grip- and limb tone (5/57), decreased abdominal tone (2/57), decreased plantary reflex (2/57), blood around the nose (2/57) and piloerection (3/57). Decreased activity (score -2;-3), abnormal gait (score +2,+3) decreased righting reflex (score -2,-3,-4), decreased grip- and limb tone (score -1,-2,-3,-4) decreased body tone (score -1;-2,-3) and piloerection (score +1) occured all animals. Incoordination (score +2,+3), blood around the eyes (score +2,+3), decreased plantary reflex (score -1, -2) and blood around the nose (score +2,+3) were observed in two animals (No.3342, 3350)
These symptoms were observed between Day 1 and Day 2.

In group 2 treated with 2000 mg/kg dose clinical sign of reaction comprised of incoordination (1 cases out of 57 observations), decreased righting reflex (4/57), decreased grip- and limb tone (4/57), decreased body tone (4/57), decreased plantary reflex (4/57) and blood around the eyes (1/57). Decreased righting reflex (-1,-2), decreased grip- and limb tone (score -1,-2,), decreased body tone (score -1,-2) and decreased plantary reflex (score -1,-2) occured in two animals (No.:3394, 3396). Incoordination (score +2) and blood around the eyes (score +2) were observed in animal No.: 3394.
These symptoms were observed between Day 1 and Day 3. Animal No.: 3378 was free of symptoms during the observation period.

Body weight:

The mean body weight of the animals corresponded to their species and age throughout the study.

Gross pathology:

All animals survived until the scheduled necropsy on Day 15.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute toxic class method (OECD Guideline 423.) was carried out involving a stepwise procedure with the use of 2000 mg/kg bw dose. The test item is ranked into classes of Globally Harmonized Classification System as GHS category 5 since the LD50 (mg/ kg bw) is above 2000.