Disodium 4-amino-3,6-bis[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxynaphthalene-2,7-disulphonate

Endpoint summary

Description of key information

NOAEL oral rat > 2000 mg/kg

NOAEL inhalation rat > 5000 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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Firma Charles River Wiga Gmbh Deutschland, Sulzfeld
- Weight at study initiation: The weights of the male rats are: 461,2 g ± 23,3 g. The weights of the female rats are: 275,2 g ± 11 g.
- Fasting period before study: 22 h
- Housing: 3 animals per cage Type 3 Makrolon
- Diet: Altromin 1326 ad libitum
- Water: tap water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20,5 - 22,5
- Humidity (%): 25-40
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

600

Doses:

Single dose: 2000 mg/kg

No. of animals per sex per dose:

6 male and 6 female

Control animals:

yes

Statistics:

Not applicable.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No mortality

Clinical signs:

other: Diarrhoea occurred in 3 male rats 5 hours after exposure to the test substance. Discoloration of the faeces occurred 5 day after the exposure.

Gross pathology:

No damage to any organs.

Other findings:

The exposure to the tested substance did not influencing the behaviour of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The tested item was found to be non toxic for oral exposure with a LD50 > 2000 mg/kg bw.

Executive summary:

The acute oral toxicity was assessed following the OECD 401.

The results show no toxicity with a LD value > 2000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Velaz Prague
- Age at study initiation: 8 weeks
- Supplier certification: declared that animals had not the external and internal parasites, pathogenic microorganisms, viruses and fungi.
- Housing: after transporting the animals were placed in polypropylene plastic nursery containers T4 (550 x 320 x 180 mm Velaz Prague), five animals each sex separately.
- Acclimation period: one week
- Diet: fed with granulated mixture ALTROMIN 1320 (Velaz Prague) in a dose of 12 g per animals each day
- Water: drinking water without restrictions

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C
- Humidity: 40-60 %
- Photoperiod: 12 hours of light and 12 hours of darkness

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

head only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: samples were dosed by WDFU (Wright Dust Feed Unit MK2, GA 4170, L. Adams, LTD., London) diredtly into an inhalation chamber, where they were dispersed through the spray; the atmosphere inside the inhalation chamber was homogeneous.
- Method of holding animals in test chamber: animals were fixed in a glass tube of diameter 60 mm so that the face of the glass cylinder to interfere with the vertical axis - 250 mm in diameter- in which there was a linear dynamic atmosphere exchange at 8 l/min.
- Source and rate of air: compressed air was taken from the central distribution Synthesia. Air flow was measured continuously throughout the exposure.

TEST CONDITIONS
- The average air flow apparatus: 0.480 m3/h
- The temperature inside the apparatus: 24°C
- Relative humidity inside the apparatus: 45%
- Actual concentration: 4.91 mg/L air

TEST SUBSTANCE
- Particle size of the test substance: :
diameter 1-4: 364.8 ± 5.86 (89.04 %);
diameter 4-10: 35.3 ± 2.48 (8,62 %);
diameter 10-20: 8.2 ± 0,64 (2.00 %);
diameter >20: 1,4 ± 0.34 (0.34 %)

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

4.91 mg/l air

No. of animals per sex per dose:

5 males and 5 non-pregnant females.

Control animals:

yes

Details on study design:

- Immediately after application the animals were removed from tube, placed in breeding containers and observed.
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: before exposure and after 7 and 14 day
- Other examinations performed: clinical diagnosis was focused on the observation of nutritional status (food intake and water), the appearance of hair and skin, visible mucous membranes, mental activity, somatomotor activity, response to stimuli, focusing on sensitization and reactivity, lacrimation, functional assessment of the respiratory, circulatory, digestive and urogenital apparatus.
- After 14 days the animals were killed and an autopsy was performed. At autopsy were examined macroscopically thoracic and abdominal organs and lungs, trachea, larynx, thymus, liver, spleen and kidneys.

Statistics:

No data

Sex:

male/female

Dose descriptor:

LC0

Effect level:

ca. 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

During the test and during the observation period (14 days) there was no mortality of the tested animals.

Clinical signs:

other:

Body weight:

The increase of body weight of animals tested was in according to the trend of increase of control animals.

Gross pathology:

Immediately thereafter after the 14-day observation period the animals were killed . After an autopsy was performed, focusing on the overall examination of the body surface and orifices, and macroscopic examination of the thoracic, abdominal and selected organs - trachea, lungs, heart, thymus, liver, spleen and kidneys.

In one male was present a diffuse pulmonary congestion.
In females were observed different effect:
- Female 1: numerous punctate hemorrhages on the right lung. No other pathological anatomical findings.
- Female 2: No pathological anatomical findings.
- Female 3: sharply defined deposits a small pea size founded in the right lung. In the left lung were observed edema.
- Female 4: grey-blue sharply defined deposits with size from pinehead to pea were observed in both lungs.
- Female 5: Similar findings has been identified as female 4.

Other findings:

Potential target organs: Trachea , lungs, heart,thymus, liver, spleen and kidneys.

Observations during the exposure:

Immediately after application the animals were removed from tubes, placed in breeding containers and observed.

- 60 min after application:

animals presented the same symptoms of intoxication of the first observation

- 120 min after application:

animals presented the same symptoms of previous observations

- 2 days after application:

stains of the test-compound on the hair, and brown-black colored (color of dark beer) urine of all of testing animals was observed. No other clinically detectable health disorders.

- From 3 to 4 days after application:

animals presented the same symptoms of previous days

- From 5 to 14 days after application: during this period there were no clinical signs of intoxication.

Interpretation of results:

GHS criteria not met

Conclusions:

A sample of Direct Black 19 were tested for limit test for acute inhalation toxicity. During the exposure or during the 14-day observation period there was no death of animals tested.
The results showed some effects that could be considered as a result of the strong irritating effects of the tested substance.

Executive summary:

Direct Black 19 was tested for acute inhalation toxicity. During exposure and during the 14-day observation period there was no mortality of experimental animals.
The effects observed during the expsoure, black fluid from eyes and nasal cavity, and after the macroscopic examination, pathological changes in the lungs (pulmonary congestion, haemorrhages in the lungs, pulmonary edema and major lobular pneumonia) can be considered as a result of strong irritating effect of the tested substances on exposed mucous upper and lower respiratory tract.
Given that these pathological changes observed in the lungs, predominantly in females (one male was found only pulmonary congestion, other males were no pathological changes), it can be considered an increasing of the sensitivity of the female sex.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 000 mg/m³ air

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No Adverse Effect level for acute toxicity oral is LD50 > 2000 mg/kg.

No Adverse Effect level for acute toxicity inhalation is LD50 > 5000 mg/m3.

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one offour toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be more than 2000 mg/kg body weight and the LD50 for inhlataion is > 5000 mg/kg bw therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity.

No classification for acute toxicity is warranted under Regulation 1272/2008.