Reaction product of disodium 4,4'-dinitrostilbene-2,2'-disulphonate, p-[(4-amino-2,5-xylyl)azo]benzenesulphonic acid, 3-[(4-amino-2-methoxyphenyl)azo]-4-hydroxybenzenesulphonic acid, copper (II) sulfate

Endpoint summary

Description of key information

LD50(oral) = 2232 mg/kg b.w

LC0(inhalation) = 2.10 mg/l

LD0(dermal) = 2700 mg/kg b.w

Key value for chemical safety assessment

Acute toxicity: via oral route

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Value:

2 232 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:

From April 21, 1978 to May 5, 1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Principles of method if other than guideline:

4-Hour Acute Inhalation Toxicity in Rats in Accordance With the Regulations as Defined in the Protocol Submitted by Ciba-Geigy. Ten laboratory rats (5 males & 5 females) are exposed in a 40 liters (36 x 36 x 31 cm) glass exposure chamber, to the test substance via the inhalation route at an atmospheric concentration of 2.34 ± 0.26 mg/l for 4 hours.

GLP compliance:

no

Test type:

fixed concentration procedure

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: no data- Weight at study initiation: 245 - 324 g- Housing: The animals were housed, individually, in suspended wire bottom cages- Diet: ad libitum- Water: ad libitumENVIRONMENTAL CONDITIONS- Temperature (°C): 23 ± 1 °C- Humidity (%): 45-55 %- Photoperiod (hrs dark / hrs light): 12 hours cycle dark/light

Route of administration:

inhalation: dust

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTIONThis test was conducted in a 40 l (36 x 36 x 31 cm) glass exposure chamber. The sides and the bottom of the chamber had centred holes (3-4 cm in diameter) to allow access to the chamber for testing and exhaust of the atmosphere. The port in the bottom of the chamber was centred over a 10 cm hole in a wooden platform. A funnel (8.5 cm in diameter) was brought from the underside of the platform through the hole and centred on the port in the bottom of the chamber. Dynamic air flow within the chamber was maintained by connecting the funnel to a vacuum pump for continuous changing of the chamber atmosphere.- Exposure chamber volume: 40 l (36 x 36 x 31 cm) glass exposure chamber.- Source and rate of air: Dynamic air flow within the chamber was maintained by connecting the funnel to a vacuum pump for continuous changing of the chamber atmosphere.- System of generating particulates/aerosols: The test substance was generated as a dust using a 3-neck, round-bottom, 250 ml Pyrex flask. A stirring mechanism was placed through the center neck of the flask and an air line through one of the side necks. The third neck was connected by a glass elbow (which had an air vent to allow flushing) to the chamber. The dust was introduced into the chamber through a side port. A piece of rubber was taped over the outer area around the hole and a vertical slit made in the rubber to allow the entrance of the glass elbow from the dust generator. Constant flow of material was maintained by a calibrated flowmeter connected between the air line and the generating apparatus. In all instances, the air flow was maintained at or above 0.5 liter of air per minute per rat. - Method of particle size determination: Particle size determinations were made using a Cascade Impactor- Treatment of exhaust air: filter holder was attached to a vacuum pump which was regulated to exhaust 1.0 liter of air per minute from the chamber through the filterTEST ATMOSPHERE- Brief description of analytical method used: Measurement of the atmospheric concentration of test substance in the chamber was achieved using a Gelman Model 1235 stainless steel filter holder containing a pre-weighed glass fiber filter (Gelman type AE-47 mm).

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

2.34 ± 0.26 mg/l

No. of animals per sex per dose:

5 per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days- Frequency of observations and weighing: 4 times for clinical signs. All animals were weighed prior to the initiation of the exposure and on days 1, 7 and 14 following exposure. - Necropsy of survivors performed: yes- Other examinations performed: clinical signs, body weight, organ weights, histopathology

Sex:

male/female

Dose descriptor:

LC0

Effect level:

ca. 2.34 mg/L air (analytical)

Based on:

test mat.

95% CL:

ca. 0.26

Exp. duration:

4 h

Key result

Sex:

male/female

Dose descriptor:

LC0

Effect level:

ca. 2.106 mg/L air (analytical)

Based on:

act. ingr.

95% CL:

ca. 0.26

Exp. duration:

4 h

Mortality:

no mortality observed

Clinical signs:

other: abnormal respiration was noted during the first 2 hours post-exposure. This condition was not noted 4 hours post-exposure and animals appeared normal for the remainder of the observation period.

Body weight:

Eight animals (4 males and 4 females) showed slight weight losses on day one; however, all animals were gaining weight normally for the remainder of the observation period.

Gross pathology:

The results of the gross pathological examinations showed that organs of all rats were within normal limits.

Interpretation of results:

GHS criteria not met

Remarks:

not classified according to CLP Regulation (EC n. 1272/2008)

Conclusions:

The test substance shows no adverse effect at concentration of 2.34 mg/l (2.10 mg/l based on active ingredient).

Executive summary:

The substance has been tested for acute toxicity via inhalation route. Ten laboratory rats (5 males and 5 females) initially weighing between 245 and 324 grams, when exposed to test substance via the inhalation route at an atmospheric concentration of 2.34 ± 0.26 mg/l for 4 hours, exhibited no observable adverse clinical signs during the exposure. Abnormal respiration was observed in all animals during the first 2 hours post-exposure. However, all animals appeared normal when observed 4 hours after the exposure. No abnormalities were noted during the 14-day post-exposure period, therefore the LC0 is 2.34 mg/l (2.10 mg/l based on active ingredient). The concentration stated above was the maximum attainable aerosol concentration of the test material under the experimental conditions. The average mass median particle diameter was 8.04 ± 1.62 µm. Gross pathological observations showed organs of all animals to be within normal limits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Value:

2 106 mg/m³

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From April 17, 1978 to May 1, 1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 434 (Acute Dermal Toxicity - Fixed Dose Procedure)

Deviations:

not specified

GLP compliance:

no

Test type:

fixed dose procedure

Species:

rabbit

Strain:

New Zealand White

Sex:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Ace Animals, Boyertown, Pennsylvania- Weight at study initiation: 2.0 and 3.1 kgThe animals were fed, housed and maintained in accordance with standard laboratory procedures

Type of coverage:

open

Vehicle:

not specified

Details on dermal exposure:

TEST SITE- % coverage: 30 % of each animal's skin surfaceREMOVAL OF TEST SUBSTANCE- Washing (if done): yes , with warm water- Time after start of exposure: 24 hours after the star

Duration of exposure:

24 hours

Doses:

3000 mg/kg bw

No. of animals per sex per dose:

3 per sex per dose

Control animals:

yes, concurrent no treatment

Details on study design:

- Duration of observation period following administration: 14 days - Frequency of observations and weighing: daily- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD0

Effect level:

ca. 3 000 mg/kg bw

Based on:

test mat.

Key result

Sex:

male/female

Dose descriptor:

LD0

Effect level:

ca. 2 700 mg/kg bw

Based on:

act. ingr.

Clinical signs:

There were no adverse clinical signs observed in any of the six animals throughout the study

Body weight:

Individual body weight fluctuations presented no consistent pattern of significance.

Gross pathology:

Necropsies revealed all organs to be within normal limits.

All animals exhibited erythema and edema on day one. On day 2, two animals showed oedema and two animals showed erythema. On the 3rd day of scoring, two animals displayed oedema and one animal displayed erythema. This lasted until day four. On day 5 and 6, one animal showed oedema and one animal showed erythema. By day 7, all animals were normal and remained so throughout the study.

Interpretation of results:

GHS criteria not met

Remarks:

not classified according to CLP Regulation (EC n. 1272/2008)

Conclusions:

The test substance at a dose level of 3000 mg/kg bw, produced oedema and erythema up to day 6, but no mortality, therefore the LD0 = 3000 mg/kg bw (2700 mg/kg b.w based on active ingredient)

Executive summary:

New Zealand albino rabbits, when exposed dermally to the test substance at a dose level of 3000 mg/kg bw, corresponding to 2700 mg/kg bw based on active ingredient, produced oedema and erythema up to day 6, but no mortality. All gross necropsies were within normal limits. Within 7 days of observation period all animals were normal and remained so throughout the study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 700 mg/kg bw

Additional information

Acute oral toxicity:

The key study made in 1974 was conducted with similar substance 1 and its concentration is 72 % . The substance shows no mortality at any concentrations and the signs of toxicity were: sedation, dyspnoea, exophthalmos, curved position and ruffled fur. These symptoms become more accentuated with the dose increased. The calculated LD50 is more than 2232 mg/kg b.w. based on active ingredient.

The supporting study made in 1990 was a short abstract, but the purity of the substance was 85 %, the composition of the tested batch is known and the curve dose/response was good described. The substance shows an LD50 = 7380.55 mg/kg b.w. No information about methods and signs of toxicity.

In the study conducted on 1973, the substance shows slight signs of toxicity by oral route, LD50 = 1837.5 mg/kg bw based on active ingredient. The rats show the following signs: ditto, ataxia, hypoventilation. The purity of the test substance is around 75 %, no data is specified by the customer about 25% of the test product. The 25 % of the substance could influence the toxicity of the experiment, therefore this is the main reason for disregarding the study.

The key study and the supporting study show a determinate behavior. In the key study it is not seen mortality at all concentrations, in the study tested in 1990 the mortality begin at concentration equal to 7943 mg/kg b.w and the 100 % of mortality was seen at 12590 mg/kg b.w. This curve dose/response support the absence of mortality seen in the key study. For cautelative manner the LD50 = 2232 mg/kg b.w it could be used for assessment of acute administration by oral route.

Acute inhalation toxicity:

The substance tested for acute toxicity by inhalation route does not show signs of toxicity , only abnormal respiration immediately after the acute dose administration. After the next two hours and since the end of post exposure period, no signs of toxicity were observed. The value of lethal concentration used in the experiment is 2.34 mg/l (2.10 mg/l based on active ingredient), it is below the trigger value for classification (5 mg/l), but no signs of mortality were observed during the experiment, therefore the value of 2.34 mg/l is not valid as LC50.

 

Acute dermal toxicity

The test substance administered by dermal route shows signs of toxicity as erythema and oedema to all tested animals. After 7 days of observation period all animals return to initial conditions and remain so throughout the study. No animals die during the acute administration of 2700 mg/kg bw and during the observation period.

Justification for classification or non-classification

According to the CLP Regulation (EC n. 1272/2008) the acute toxicity is:

acute toxicity means those adverse effects occurring following oral or dermal administration of a single dose of a substance or a mixture, or multiple doses given within 24 hours, or an inhalation exposure of 4 hours.

After the oral administration the LD50 of test item is 2232 mg/kg b.w (based on active ingredient), .

After acute dermal and inhalation administration no lethal dose (LD50 or LC50) has been derived. Taking into account that no effect at all has been reported at 2.10 mg/L for inhalation , and 2700 mg/kg for dermal it can be assumed that :

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

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

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