Disodium 6-hydroxy-5-[[4-[[4-(phenylamino)-3-sulphonatophenyl]azo]naphthyl]azo]naphthalene-2-sulphonate

Endpoint summary

Description of key information

According to the study results (acute toxic class method) the value of LD50 of the test substance, Acid Black 26, for female rats is higher than 2000 mg/kg of body weight.

A limit test for acute inhalation toxicity was performed according to OECD TG 403 methodology. The results of the test revealed the P-deposition with test substance particles but did not reveal any adverse effects on the health of the experimental animals and the value of LD50 for male/female rats is higher than 5200 mg/m³.

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:

12.10. – 27.10.2015

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Remarks:

Wistar Han, monitored quality

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: breeding farm VELAZ s.r.o., Únětice, Czech Republic, RČH CZ 21760118- Age at study initiation: 6 - 7 weeks- Weight at study initiation: 171 - 183 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 standard diet for experimental animals ad libitum- Water (e.g. ad libitum): drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 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:

water

Details on oral exposure:

VEHICLEAqua pro iniectione - Lot/batch no.: 1505050287 (expiration 05/2017), producer Ardeapharma Ševětín, 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 in vehicle (aqua pro iniectione) and resulting suspension was administered to the stomach by tube.

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 day- Necropsy of survivors performed: yes- Other examinations performed: body weight, clinical signs, 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:

Black coloured faeces first day after application in all animalsdiarrhoea (at first and second day after application in all six females). No other 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 animals. Weight increments were adequate to species, sex and age of animals in experiment.

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. The test substance caused black coloured faeces first day after application in all animals. Diarrhoea was observed first and second day after application in all six females. 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 26, 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 26, after a single oral administration to Wistar rats.

The testing was performed according to the 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 evaluation of results was performed according to the diagrams given in Annex 1 D (Starting dose 2000 mg/kg bw).

The test substance was administered in a single dose as solution in vehicle (aqua pro inectione), given orally via gavage to female Wistar rats.

On the basis of information about no toxicity of the test substance, the starting dose of 2000 mg/kg of body weight was used in the study. 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 2000 mg/kg caused no death of animals. No serious clinical signs of intoxication were detected during the whole study in all six animals administered by the dose 2000 mg/kg/body weight. The test substance caused black coloured faeces first day after application in all six animals. Diarrhoea was observed first and second day after application in all six females.

No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test substance, Acid Black 26, 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

Quality of whole database:

Reliability 1

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14.09. – 28.09.1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

OECD Guideline for Testing of Chemicals, 1981

Deviations:

no

GLP compliance:

yes

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: breeding farm VELAZ Praha- Females (if applicable) nulliparous and non-pregnant: yes- Microbiological status of animals, when known: delivered with veterinary attest “QUALITY STATUS” – no parazites, pathogen microorganisms, viruses or mold- Age at study initiation: 8 weeks- Weight at study initiation: tested animals; males: 170.8 ± 7.12 g; females: 161.2 ± 4.22 g- Housing: 5 animals in 1 polypropylene cage T4 type (VELAZ), separated male/female - Diet (e.g. ad libitum): granulated mixture Altromin 1320 (VELAZ Praha); dose: 12g/animal/day- Water (e.g. ad libitum): ad libitum- Acclimation period: 1 week as minimumENVIRONMENTAL CONDITIONSAutomatically controlled temperature, humidity and lighting - Temperature (°C): 22±3 - Humidity (%): 40 – 60% - Air changes (per hr): not specified- Photoperiod (hrs dark / hrs light): 12 /12

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

head only

Vehicle:

air

Remark on MMAD/GSD:

see tables in attached pdf file

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION - Exposure apparatus: chamber "head only" type - Exposure chamber volume: dynamic air flow 10 L/min - Method of holding animals in test chamber: glass tubuses of 60 mm diameter - Source and rate of air: central source of pressure air, flow rate was measured continually - System of generating particulates/aerosols: WDFU (Wright Dust Feed Unit MK2, GA 4170, L.Adams, London) - Method of particle size determination: microscopically - Temperature, humidity, pressure in air chamber: 24 °C and humidity 45% (measured in middle part of chamber); negative pressure 98.07 Pa TEST ATMOSPHERE - Brief description of analytical method used: The samples of atmosphere inside the chamber were taken at a rate of 4L/min, total amount 10 L of air/sample.The air samples were passed through a collecting cartridge filled with filter material and the concentration of the test substance inside the inhalation chamber was determined gravimetrically. - Samples taken from breathing zone: yes VEHICLE - Composition of vehicle (if applicable): air - Concentration of test material in vehicle (if applicable): 5.2 mg/L of air TEST ATMOSPHEREBefore starting the exposure a particle size of the initial sample was measured microscopically. The size of the majority of particles was between 2- 4 µm. (see tables in attached pdf file)

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5.2 mg/L of air

No. of animals per sex per dose:

5 male/5 female

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days - Frequency of observations and weighing: immediately after end of exposure, after 1st and 2nd hour, then dailybody weight was measured immediately before exposure, then on the 7th and 14th day - Necropsy of survivors performed: yes - Other examinations performed: clinical signs, body weight, food and water intake, appearance of hair, skin and visible mucous membranes, somatomotor and mental activity, reaction to stimuli, focusing on sensibility and reactivity, lacrimation, functional assessment of the respiratory, circulatory, digestive and urogenital system

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.2 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

no

Clinical signs:

other: During exposure examinationAfter 20 minutes exposure: black discharge from the nasal cavity. No motor disorders of the body have been observed.During post exposure period0, 60 and 120 min post exposure: black discharge from the nasal cavity and eyes, tro

Body weight:

see attached document Midlonová čerň VL_acute toxicity inhal_tables

Gross pathology:

The results of the limit test revealed the P-deposition with test substance particles but did not reveal any adverse effects on the health of the experimental animals. Differences in relation with the sex of the animals were not recorded. Reactions and autopsy findings were the same.

Interpretation of results:

GHS criteria not met

Conclusions:

A limit test for acute inhalation toxicity with substance Midlonová čerň VL was performed. During the exposure and during the 14-day observation period there was no death among tested animals.Clinical examination of health condition suggests a mechanical irritation due to test substance on exposed mucous membranes of the upper respiratory tract, reduced airway patency and insufficient oxygen supply to vital organs.The results of the pathological-anatomical examination revealed a P-deposition of inhaled material particles without local or general response of the organism of tested animals.Differences in relation with the sex of the animals were not recorded. Reactions and autopsy findings were the same.The results of the limit test revealed the P-deposition with test substance particles but did not reveal any adverse effects on the health of the experimental animals.

Executive summary:

A limit test for acute inhalation toxicity with substance Midlonová čerň VL was performed. The test was performed according to OECD TG 403 methodology. Experimental animals (10 rats Wistar) were exposed to an aerosol inhalation of the test substance powder in air for 4 hours in the "head only" inhalation chamber. Actual test substance concentration was measured by a gravimetric method. Prior to exposure, the particle size of the initial sample was measured microscopically. The particle size was predominantly within the range 2- 4 µm. Particle size is one of the limiting factors in the deposition of inhaled material in different areas of the respiratory apparatus.

During the exposure and during the 14 day observation period there was no death among tested animals.

Signs of intoxication were clinically determined and give an evidence of mechanical irritation of dust aerosol on exposed mucous membranes of the upper respiratory tract, reduction in airway patency and insufficient oxygen supply to vital organs.

Pathological examination demonstrated a focal infiltration of pulmonary tissues by test substance particles.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 200 mg/m³

Quality of whole database:

Reliability 1

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In accordance with column 2 of REACH Annex VIII, Acute toxicity by dermal route (required in section 8.5.3.) does not need to be conducted. The acute toxicity by inhalation route is available.