Reaction mass of 2-[(4-methyl-2-nitrophenyl)diazenyl]-3-oxo-N-phenylbutanamide and 2-[(4-nitrophenyl)diazenyl]-3-oxo-N-phenylbutanamide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11.09. – 27.09.2017

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

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

2008

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:

2-[(p-nitrophenyl)azo]acetoacetanilide

EC Number:

216-754-0

EC Name:

2-[(p-nitrophenyl)azo]acetoacetanilide

Cas Number:

1657-16-5

Molecular formula:

C16H14N4O4

IUPAC Name:

2-[(4-nitrophenyl)diazenyl]-3-oxo-N-phenylbutanamide

impurity 1

Reference substance name:

Unknown impurities

Molecular formula:

Not available

IUPAC Name:

Unknown impurities

additive 1

Reference substance name:

Resin acids and Rosin acids, aluminum salts

EC Number:

263-075-0

EC Name:

Resin acids and Rosin acids, aluminum salts

Cas Number:

61789-65-9

Molecular formula:

Not available - UVCB substance

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Remarks:

monitored quality

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: breeding farm VELAZ s.r.o., Lysolaje, Czech Republic, RČH CZ 21760118- Age at study initiation: 8 weeks- Weight at study initiation: Step No.1 average 139.3 g; Step No.2 average 156.7 g- Housing: animal room with monitored conditions – 3 animals of one sex in one plastic breeding cage with sterilized shavings of soft wood- Diet (e.g. ad libitum): pelleted diet for rats and mice Altromin ad libitum- Water (e.g. ad libitum): drinking water ad libitum quality corresponding to Regulation No. 252/2004 of Czech Coll. of Law- Acclimation period: 5 daysENVIRONMENTAL CONDITIONS- Temperature: 22 ± 3°C, permanently monitored- Humidity: 30 – 70 %, permanently monitored- Photoperiod: 12 hour light/12 hour dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLEolive oil - Lot/batch no.: 8002182001 (expiration 07/2018), producer Dr. Kulich Pharma, s.r.o., Czech RepublicDOSING: On the basis of information about no toxicity of the test substance, the starting dose of 2000 mg/kg body weight was used. Because this dose caused no death of females, same dose of 2000 mg/kg level was sequentially applied for confirmation (application with time distance 24 hours) to group of 3 females. No death of animals was observed in this group of 3 females.PREPARATION AND APPLICATION OF THE TEST SUBSTANCEImmediately before application the test substance was weighed, mixed with vehicle (olive oil) and resulting suspension was administered to the stomach by tube. All prepared suspensions of the test substance in olive oil were mixed by magnetic stirrer during administration.

Doses:

2000 mg/kg (first step)2000 mg/kg (confirmation)

No. of animals per sex per dose:

3 animals per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days- Frequency of observations and weighing: 1st day: twice (30 minutes and 3 hours after application), 2nd day: twice (in the morning and in the afternoon), thereafter days: once a dayClinical signs: observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system. The results of the observations were recorded on special data sheets.- Necropsy of survivors performed: yes- Other examinations performed: nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded on special data sheets.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death of animals.

Clinical signs:

No clinical signs of intoxication were observed after dosing 2000 mg/kg/body weight.

Body weight:

Body weight was recorded and weight increments were calculated in all surviving animals at the end of study. Weight increments were adequate to species, sex and age of animals in experiment. No reduction of body weight was observed

Gross pathology:

No pathologic macroscopic changes were diagnosed during pathological examination.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings at the end of study. The test substance administered at the dose of 2000 mg/kg caused no death of females. No serious clinical signs of intoxication were detected at this dose during whole study. No pathologic macroscopic changes were diagnosed during pathological examination.According to the study results the value of LD50 of the test substance, Pigment Yellow 4, for female rats is higher than 2000 mg/kg of body weight.

Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance, Pigment Yellow 4, after a single oral administration to Wistar Han rats.

The testing was performed according Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

The test substance was administered in a single dose as a suspension in vehicle (Olive oil), given orally via gavage to female Wistar rats. The volume of administered suspension was 1 ml/100 g body weight of animals.

The dose level of 2000 mg/kg was used as the starting dose.

The dosing was performed sequentially in three groups of three females: group No. 1 - first step using the starting dose of 2000 mg/kg of body weight and group No. 2 – second step using the same dose 2000 mg/kg.

The test substance administered at the dose of 2000 mg/kg caused no death of animals. No serious clinical signs of intoxication were detected during whole study. No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test substance Pigment Yellow 4, for female rats is higher than 2000 mg/kg of body weight.