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

Endpoint summary

Description of key information

According to the acute oral toxicity 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. No serious clinical signs of intoxication were detected at this dose during whole study. No pathologic macroscopic changes were diagnosed during pathological examination.

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.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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

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

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

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.

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Reliability 1

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

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

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

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

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.

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Reliability 1