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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30.10. – 16.11.2017

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.3 (Acute Toxicity (Dermal))

Version / remarks:

Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008

Deviations:

no

GLP compliance:

yes

Test type:

standard acute 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:

Wistar CRL, monitored quality

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Breeding farm VELAZ s.r.o., Lysolajské údolí 15/53, Czech Republic, RČH CZ 11760500; with certificate of good health condition - Age at study initiation: not specified- Weight at study initiation: average F 216.2 g, M 249.9 g - Fasting period before study: not specified- Housing: individually in plastic breeding cage with sterilized shavings of soft wood, monitored conditions - Diet (e.g. ad libitum): pelleted diet ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: at least 5 days ENVIRONMENTAL CONDITIONS- Temperature: 22 ± 3°C, permanently monitored- Humidity: 30 – 70 %, permanently monitored- Photoperiod: 12 hour light/12 hour dark

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Remarks:

moistened with water

Details on dermal exposure:

TEST SITE - Area of exposure: 6 × 6 cm on the shaved back of animals (aprox. 10% of the body surface) - Type of wrap if used: gauze, plaster REMOVAL OF TEST SUBSTANCE - Washing (if done): wiped off with water - Time after start of exposure: 24 h TEST MATERIAL - Amount(s) applied (volume or weight with unit): 2000 mg/ kg bw - For solids, paste formed: no, applied in delivered form, moistened with water to ensure good contact with the skin

Duration of exposure:

24 h

Doses:

2000 mg/ kg bw

No. of animals per sex per dose:

5 males + 5 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days - Frequency of observations and weighing: Body weight: before application, 8th and 15th day of study Mortality: dailyClinical signs: dailyAfter application the animals were observed individually – at the first day: twice (30 minutes and 3 hours after application), at the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days. 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: yesPathological examination:15th day of studyAll test animals surviving to the end of study were sacrificed on the 15th day by diethyl ether narcosis and gross necropsy was carried out. Nutritional state, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 000 mg/kg bw

Based on:

test mat. (total fraction)

Mortality:

No death of animals was observed during the 14-day observation period.

Clinical signs:

No clinical signs of intoxication were observed.

Body weight:

In males the weight increments were adequate to species and age of animals in the experiment. In three females only the slight decrease of body weight was observed during the first week after application.

Gross pathology:

No macroscopic changes were diagnosed during pathological examination in all animals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance toxicity was evaluated on the basis of mortality, body weight changes, and clinical signs of toxicity during the observation period and necropsy findings at the end of study. The test substance applied on skin at the dose of 2000 mg/kg of body weight did not cause death of any animals. No clinical signs of toxicity were observed during the entire study. No macroscopic changes were diagnosed during pathological examination. According to the results of this study, the LD50 value (dermal) of the test substance, Pigment Yellow 4, for rats of both sexes is higher than 2000 mg/kg of body weight.LD50> 2000 mg/kg

Executive summary:

The test substance, Pigment Yellow 4, was tested for acute dermal toxicity using Wistar rats.

Testing was performed according to Method B.3 - Acute toxicity (Dermal), Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008

The study was performed as a limit test: two groups of animals – 5 males and 5 females at a dose of 2000 mg/kg of body weight. The pre-test was performed with 1 male and 1 female from each group. After the end of exposure of these pilot animals, the other animals of the group were dosed.

The test substance (moistened with water) was applied on the shaved skin (approx. 10 % of body surface) of the test animals and held in place by a semi-occlusive dressing for 24 hours.

The test animals were observed daily for 14 days after ending of exposure of the test substance, sacrificed, and necropsy for macroscopic examination of the organs was performed.

The test substance applied at a dose of 2000 mg/kg of body weight did not cause death of any animals. No clinical signs of toxicity were observed during the entire study. No macroscopic changes were diagnosed during pathological examination.

According to the results of this study, the LD50 value (dermal) of the test substance, Pigment Yellow 4, for rats of both sexes is higher than 2000 mg/kg of body weight.