Disodium [3-[4,5-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonato(3-)][1-[[2-hydroxy-5-(phenylazo)phenyl]azo]-2-naphtholato(2-)]chromate(2-)

Endpoint summary

Description of key information

LD50 (male/female)= 5437 mg/kg bw. 

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:

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 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

In order to determine the dose/response relationship of the test substance, groups of 5 male and 5 female rats were treated with various single doses of the compound, diluted in carboxymethylcellulose by oral intubation. Symptoms and mortality after administration were recorded during an observation period of 14 days.

GLP compliance:

no

Remarks:

pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Tif RAI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age: 6 o 7 weeks
- Weight at study initiation: 160 g to 180 g
- Housing: housed in groups of 5 in macrolon cages (Size 3)
- Fasting period before study: animals were starved during one night before starting the treatment
- Diet and water: animals were fed a standard diet of Nafag ad libitum.


ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1 °C
- Humidity: approximately 50 %

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

2 %

Details on oral exposure:

- Concentration in vehicle: 30 % or 40 %

Doses:

2780, 4640, 6000, 7750 and 10000 mg/kg bw

No. of animals per sex per dose:

Groups of 5 male and 5 female rats

Control animals:

no

Details on study design:

Symptoms and mortality after administration were recorded during an observation period of 14 days.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 5 437 mg/kg bw

Based on:

test mat.

95% CL:

> 4 715 - < 6 270

Mortality:

Mortality was observed starting from the dose of 4640 mg/kg bw at 24 hours.

Clinical signs:

Within 2 hours after treatment the rats in all dosage groups showed sedation, dyspnoea, exophthalmus, curved position, diarrhoea and ruffled fur. Sedation, diarrhoea and ruffled fur became more accentuated as the dose was increased.

Gross pathology:

No substance related gross organ changes were seen in the killed animals.

Dose (mg/kg)	Concentration (%) of formulation	Number of animals		Died within
				1 hr		24 hrs		48 hrs		7 days		14 days
		Male	Female	Male	Female	Male	Female	Male	Female	Male	Female	Male	Female
2780	30	5	5	0	0	0	0	0	0	0	0	0	0
4640	40	5	5	0	0	1	0	2	1	2	2	2	2
6000	40	5	5	0	0	1	3	1	4	1	4	1	4
7750	40	5	5	0	0	2	5	2	5	4	5	4	5
10000	40	5	5	0	0	4	4	5	5	5	5	5	5

The LD50 was calculated by probit analysis method (Goulden A., Methods of Statistical Analysis, John Wiley and Sons, 1960, 3rd printing, pages 404-408).

Interpretation of results:

other: Not classified according to the CLP Regulation

Conclusions:

The acute oral LD50 of the susubstance in rats of both sexes observed over a period of 14 days is 5437 (4715-6270) mg/kg bw.

Executive summary:

The oral acute toxicity study was performed according to an internal method: groups of 5 male and 5 female rats were treated with various single doses of the compound, diluted in carboxymethylcellulose by oral intubation. Symptoms and mortality after administration were recorded during an observation period of 14 days.

The acute oral LD50 of the substance in rats of both sexes observed over a period of 14 days is 5437 (4715-6270) mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 437 mg/kg bw

Quality of whole database:

The test results/procedures cannot be subsumed under a testing guideline, but they are nevertheless well documented and scientifically acceptable.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Despite the key study was not carried out according to a OECD Guideline, but to an internal method, the test turns out to be sufficiently detailed. Groups of 5 male and 5 female rats were treated with various single doses of the compound, diluted in carboxymethylcellulose administered by oral intubation. Symptoms and mortality after administration were recorded during an observation period of 8 days.

The acute oral LD50 in rats was established to be 5437 mg/kg bw.

Justification for classification or non-classification

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

In the case of oral exposure route, the acute toxicity hazard categories and acute toxicity estimates (ATE) defining the respective categories are:

- 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 acute oral LD50 in rats was established to be 5437 mg/kg bw.

Therefore, the substance is not classified for oral acute toxicity according to the CLP Regulation (EC n. 1272/2008).