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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

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Diss Factsheets

Administrative data

acute toxicity: inhalation
Type of information:
other: experimental results on similar substance
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study well documented, meets generally accepted scientific principles, acceptable for assessment.

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 403 (Acute Inhalation Toxicity)
GLP compliance:
Test type:
acute toxic class method
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 4-amino-3,6-bis[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxynaphthalene-2,7-disulphonate
EC Number:
EC Name:
Disodium 4-amino-3,6-bis[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxynaphthalene-2,7-disulphonate
Cas Number:
Molecular formula:
disodium 4-amino-3,6-bis({4-[(2,4-diaminophenyl)diazenyl]phenyl}diazenyl)-5-hydroxynaphthalene-2,7-disulfonate
Test material form:
solid: particulate/powder

Test animals

Details on test animals or test system and environmental conditions:
- 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

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

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
head only
Details on inhalation exposure:
- 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.

- 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

- 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:
Duration of exposure:
ca. 4 h
4.91 mg/l air
No. of animals per sex per dose:
5 males and 5 non-pregnant females.
Control animals:
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.
No data

Results and discussion

Effect levelsopen allclose all
Dose descriptor:
Effect level:
ca. 5 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Dose descriptor:
Effect level:
> 5 mg/L air
Based on:
test mat.
Exp. duration:
4 h
During the test and during the observation period (14 days) there was no mortality of the tested animals.
Clinical signs:
other: The main signs were: - ca 10 minutes after exposure, emission of black non-foamed liquid from nasal cavity and eyes was observed. - Motor disorders of the body, legs and of the tails were observed.
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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
The tested item 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:
The substance 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.