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Description of key information

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. Only one study is available (GLP, Klimish score 1)

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:
25.09. - 11.10.2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: SPF breeding, VELAZ s.r.o., Lysolajské údolí 15/53, 165 00 Prague 6, Czech Republic, RČH CZ 11760500- Females nulliparous and non-pregnant: yes- Age at study initiation: 8 weeks at the application- Weight at study initiation: 175.08 -183.43 g- Fasting period before study: 20 h before aplication- Housing: animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage, shavings of soft wood- Diet: ad libitum, pelleted standard diet for experimental animals - Water: ad libitum, drinking tap water (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law- Acclimation period: 12 days ENVIRONMENTAL CONDITIONS - Temperature (°C): 22 ± 3 °C, permanently monitored - Humidity (%): 30 – 70 %, permanently monitored - Photoperiod (hrs dark / hrs light): light period 12-hour light/12 hour dark Animal supply: 14.09. 2016Experimental part of study: 26.09. - 11.10. 2016
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
DosingThe test substance was administered to the stomach by tube. The single volume of administered suspension was 1 mL/100 g of animal body weight. The starting dose was 2000 mg/kg of body weight.CLASS METHOD - Rationale for the selection of the starting dose: The dose level of 2000 mg/kg was used as the starting dose, because no toxicity of the test substance was estimated on the basis of information from Sponsor about testing of similar substances.START: 2000 mg/kg – 3 females (Step No. 1) – no deaths ►2000 mg/kg – 3 females (Step No. 2): no deaths ► END of study
Doses:
2000 mg/kg
No. of animals per sex per dose:
6
Control animals:
no
Details on study design:
- Body weight recordingAnimals were weighed before application, at the 8th day of study and at the 15th day, before euthanasia of animals. Average body weight in a group was calculated from individual body weights. Body weight increments were calculated from body weight at the start of the study, the first week and at the end of the study.- Clinical examinationAfter application the animals were observed individually: - the first day: twice (30 minutes and 3 hours after application)- 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. - Pathological examinationAll test animals survived to the end of study were sacrificed on the 15th day and gross necropsy was carried out. 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.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
no mortality
Clinical signs:
Changes of colour of faeces associated with the colour of the test substance were observed during the clinical observation in the second and the third day after the application. 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 animals. Only changes of colour of faeces associated with the colour of the test substance were observed. No other clinical signs of intoxication were observed after dosing 2000 mg/kg/body weight.Weight increments were adequate to species, sex and age of animals in experiment. No pathologic macroscopic changes were diagnosed during pathological examination.According to the study results the value of LD50 of the test substance, Acid Black 227, 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, Acid Black 227, 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 (Aqua pro iniectione), 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, because no toxicity of the test substance was estimated on the basis of the information from Sponsor.

The dosing was performed sequentially in two groups of three females: group No. 1 - using the starting dose of 2000 mg/kg of body weight. No death of females caused this dose, dose of 2000 mg/kg of body weight was sequentially applied for confirmation to group No. 2.        

The test substance administered at the dose of 2000 mg/kg caused no death of animals. Only changes of colour of faeces associated with the colour of the test substance were observed during the clinical observation from the first to the third day after the application. No other clinical signs of intoxication were detected during the whole study in all six animals administered by the dose 2000 mg/kg/body weight. 

No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test substance, Acid Black 227, for female rats is > 2000 mg/kg of body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Justification for classification or non-classification