Reaction mass of tetrasodium 3-[[4-[[4-[(6-amino-1-hydroxy-3-sulphonato-2-naphthyl)azo]-6-sulphonato-1-naphthyl]azo]-1-naphthyl]azo]naphthalene-1,5-disulphonate and tetrasodium 3-[[4-[[4-[(6-amino-1-hydroxy-3-sulphonato-2-naphthyl)azo]-7-sulphonato-1-naphthyl]azo]-1-naphthyl]azo]naphthalene-1,5-disulphonate

Endpoint summary

Description of key information

LD50, oral, rat = 5000 mg/kg bw /day (LD0)

LC50, 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

Study period:

1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

GLP compliance:

no

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

HealthySpregue-Dawley RAI.f. (SPF) (Ra25) derived rats were used. These were bred on the premises. Aged 6-7 weeks, and had an average body weight of 154 g. (male) and 127 g. (female).
Rats were caged singly and kept in a room maintained at a temperature of 21°C. Animals were subjected to 12 hours artificiale light and 12 hours darkness in each 24 hour period. A commercial autoclavable pelleted diet (Labsure CMR Rat and mouse nuts) was fed and lib. Water filtered at 0.45 micron was available at all times.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

A 25% w/v suspension of the compound in a 0.5% aqueous solution of carboxymethyl cellulose was administered as a single dose by gavage to rats which had been fasted for 18 hours, at a rate of 20ml/kg. (equivalent to 5g/kg of compound)

Doses:

5g/kg

No. of animals per sex per dose:

5 females and 5 males per dose

Control animals:

no

Details on study design:

After administration of the compound, the animals were observed for 14 days. Death and clinical symptoms were recorded. At the end of the observation period, surviving animals were killed by exsanguination under ether anaesthesia and an autopsy performed.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 5 000 mg/kg bw

Based on:

test mat.

Mortality:

no death

Clinical signs:

other: no abnormalities

Gross pathology:

dark kidney surfaces in 3/5 male and 4/5 female animals

Other findings:

Black stained urine and faeces were seen from 3 hours up to 6 days post dose. No abnormalities were seen from day 7 until the study terminated on day 14. At terminal autopsy the only abnormalities noted were dark kidney surfaces in 3/5 male and 4/5 female animals.

Interpretation of results:

other: not classigied under Regulation 1272/2008

Conclusions:

The substance was tested for acute toxicity oral follwoing OECD 420. Under the experimental conditions the substance showed LD50 (LDo= > 5000 mg/kg and did not show any toxicity signs over the 14 days period of observation

Executive summary:

The substance was tested following OECD 420. Ten rats, five males and five females, were kept and fed in the same conditions by a 25% w/v suspension of the compound in a 0.5% aqueous solution of carboxymethyl cellulose. The substance was administered as a single dose by gavage to rats which had been fasted for 18 hours, at a rate of 20ml/kg. (equivalent to 5mg/kg of compound). After administration of the compound, the animals were observed for 14 days. Deaths and clinical symptoms were recorded. No abnormalities were seen from day 7 until the study terminated on day 14. At terminal autopsy the only abnormalities noted were dark kidney surfaces in 3/5 male and 4/5 female animals. The substance can be considered as non toxic at doses ca. 5000 mg(kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 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

Study period:

September 27, 1994 to October 11, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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.
- Diet: fed with granulated mixture ALTROMIN 1320 (Velaz Prague) in a dose of 12 g per animals each day
- Water: drinking water without restrictions
- Acclimation period: one week

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 dispensing by WDFU (Wright Dust Feed Unit MK2, GA 4170, L. Adams, LTD., London) to an inhalation chamber, where they w ere 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 9 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: 0.540 m3/h
- The temperature inside the apparatus: 24°C
- Relative humidity inside the apparatus: 50%
- Actual concentration: 5.16 mg/L air

TEST SUBSTANCE
- Particle size of the test substance: :
diameter 1-4 um: 78,65%
diameter 4-10 um: 16,00%
diameter 10-20 um: 4,37%
diameter >20 um: 0.98%

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

5.16 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: at immediatly after exposure, 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.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

ca. 5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: also LC0

Mortality:

During the exposure there was no mortality of animals tested.

Clinical signs:

other: Clinical signs were due to mechanical irritant effect of aerosol dust on exposed mucous membranes and upper respiratory track. The main signs were: the swelling of the eyelids, runny eyes and nasal. Motor disorders of the forelegs and of the tails were ob

Body weight:

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

Gross pathology:

Immediately 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 and in one female were presents haemorrhages in the lungs. In both cases, these haemorrhages were located under the pulmonary pleura. In one female were found congested kidneys

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.

Motor disorders on the body,irregular breathing were recorded.

- 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:

shaggy skins, different parts of the body covered by residues of the test substance,blue urine and swollen eyelids.

- 3 days after application:

in addition to the blue color of the urine were not check for any effects .

- 4 days: apart from slight blue discoloration of urine were no clinical signs of intoxication.

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

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: not specified

Conclusions:

The substance was tested for acute toxicity by inhalation following OECD 403. Under the experimental conditions the LC50 (LC0) resulted equal to 5.16 mg/l (dust aerosol).

Executive summary:

The substance was tested for acute inhalation toxicity following OECD 403. The test was performed on 10 rats WISTAR strain, that were exposed to inhalation of dust aerosol test substance for 4 hours in the inhalation chamber. The test chemical concentration was measured gravimetric method. The size of most particles ranged from 2 to 4 µm. Particle size is one of the limiting factors in the deposition of inhaled substances in different areas of the respiratory apparatus.

During the exposure or during the 14-day observation period, there was no mortality of test animals.

Clinical signs of intoxication can be described as the result of mechanically irritating dust aerosol effect on exposed mucous membranes and upper respiratory tract (HCD).

The pathological-anatomical examination showed a infiltration of lung tissue particles of the substance tested. The measured particle size of the substance tested for acute inhalation are one order of magnitude smaller than the dimensions determined for the registered substance.

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

Justification for selection of acute toxicity – oral endpoint

Both studies are in agreement and with good reliability. The study conducted following the most updated OECD (420) has been selected as key study

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008), table 3.1.1, Acute toxicity hazard categories and acute toxicity estimates (ATE) defining the respective categories:

For Acute toxicity oral route:

Category 1: ATE <= 5 mg/kg bw

Category 2: 5 < ATE <= 50 mg/kg bw

Category 3: 50 < ATE <= 300 mg/kg bw

Category 4: 300 < ATE <= 2000 mg/kg bw

The LD50 of the test substance was determined to be greater than 5000 mg/kg bw in the chosen reference test, which is outside the above criteria. Therefore, the test substance is not classified for Acute toxicity by oral exposure.

 

For Acute toxicity inhalation route (mists, mg/l)

Category 1: ATE <= 0.05

Category 2: 0.05 < ATE <= 0.5

Category 3: 0.5 < ATE <= 1

Category 4: 1 < ATE <= 5

The LC50 of the test substance was determined to be 5 mg/l, therefore, the test substance is not classified for Acute toxicity by inhalation exposure.