Disodium 2,5-dichloro-4-(5-hydroxy-3-methyl-4-(sulphophenylazo)pyrazol-1-yl)benzenesulphonate

Endpoint summary

Description of key information

LD50 (oral) > 5000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

Groups of four Sprague-Dawley rats (two males and two females) were intubated with the sample at selected close levels, i.e. 4600, 6800, 10200, 15400 mg/kg. All doses were administered by gavage. Following oral administration of the test material, the rats were observed for the succeeding 14 days. Initial and final body weights as well as all mortalities and/or reactions displayed were recorded. A necropsy was conducted on any animal which died during the study as well as on all animals sacrificed at the end of the 14 days.

GLP compliance:

no

Remarks:

pre GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: from 150 to 227 grams.
- Housing: following oral administration of the test material, the rats were housed individually in suspended, wire-mesh cages.
- Fasting period before study: 16 hours period immediately prior to oral intubation.
- Diet: standard laboratory diet, ad libitum.
- Water: ad libitum.
- Acclimation period: all animals were kept under observation for five days prior to experimental use, during which period they were checked for general physical health and suitability as test animals.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

All doses were administered directly into the stomachs of the rats using a hypodermic syringe equipped with a ball-tipped intubating needle.
The test material was administered as a 50. 0 percent (w/v) aqueous suspension.

Doses:

4600, 6800, 10200, 15400 mg/kg

No. of animals per sex per dose:

2 males and 2 females

Details on study design:

- Duration of observation period following administration: 14 days.
- Observations and weighing: initial and final body weights as well as all mortalities and/or reactions displayed were recorded.
- Necropsy of survivors performed: necropsy was conducted on any animal which died during the study as well as on all animals sacrificed at the end of the 14 days.

Statistics:

At the end of the observation period, the acute oral median lethal dose (LD50) of each test material was calculated using the techniques of Weil, Thompson, and Thompson and Weil.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

10 200 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Standard Deviation of LD50 = ± 1200 mg/kg

Mortality:

No death occurred at 4600 and 6800 mg/kg. At 10200 2 oput of 4 rats died, while at 15400 mg/kg no rat survived.

Clinical signs:

other: Hypoactivity and ruffed fur were observed at 4600 mg/kg; hypoactivity, ruffed fur and muscular weakness at the two middle doses. At the highest tested dose hypoactivity, ruffed fur, muscular weakness and prostration were observed.

Gross pathology:

Necropsy of the animals that died revealed gastroenteritis. No gross pathologic alterations were noted among the animals sacrificed at the end of the 14-day observation period.

Mortality and Body Weight Data

Dose (mg/kg)	Animal N. and sex	Individual body weight (g)		Number Dead/Number Tested	Percent Dead
		Test day N.
		0	14
4600	1M	184	242	0/4	0
	2M	160	244
	3F	174	194
	4F	172	177
6800	5M	182	250	0/4	0
	6M	177	250
	7F	158	192
	8F	158	167
10200	9M	178	(6-22 hrs)	2/4	50
	10M	205	268
	11F	156	(6-22 hrs)
	12F	150	178
15400	13M	160	(6-22 hrs)	4/4	100
	14M	189	(6-22 hrs)
	15F	163	(6-22 hrs)
	16F	184	(6-22 hrs)

Summary of Reactions

Dose (mg/kg)	Reaction	Time of onset after dose administration	Duration of reaction
4600	Hypoactivity	1 hour	2 days
	Ruffed fur	6 - 2 2 hours	1 day
6800	Hypoactivity	1 hour	3 days
	Ruffed fur	5 hours	3 days
	Muscular weakness	5 hours	1 day
10200	Hypoactivity	1 hour	3 days
	Ruffed fur	1 hour	3 days
	Muscular weakness	3 hours	2 days
15400	Hypoactivity	1 hour	Until death
	Ruffed fur	1 hour	Until death
	Muscular weakness	1 hour	Until death
	Prostration	5 hours	Until death

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

LD50: 10200 mg/Kg bw.

Executive summary:

Method

Young albino rats of the Sprague-Dawley strain were used in order to assess the cute toxicity potential by otral route of the test item. Selected groups of four rats (two males and two females) were intubated with the sample at selected close levels, i.e. 4600, 6800, 10200, 15400 mg/kg. All doses were administered directly into the stomachs of the rats using a hypodermic syringe equipped with a ball-tipped intubating needle. Following oral administration of the test material, the rats were observed for the succeeding 14 days. Initial and final body weights as well as all mortalities and/or reactions displayed were recorded. A necropsy was conducted on any animal which died during the study as well as on all animals sacrificed at the end of the 14 days.

Results

No death occurred at 4600 and 6800 mg/kg; at 10200 mg/kg 2 out of 4 rats died, while at 15400 mg/kg no rat survived.

Hypoactivity and ruffed fur were observed at 4600 mg/kg; hypoactivity, ruffed fur and muscular weakness at the two middle doses. At the highest tested dose hypoactivity, ruffed fur, muscular weakness and prostration were observed.

Necropsy of the animals that died revealed gastroenteritis. No gross pathologic alterations were noted among the animals sacrificed at the end of the 14-day observation period.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

Healthy Sprague-Dawley were administered with a 25 % w/v solution of the compound in tap water, as a single dose by gavage at a rate of 20 ml/kg (equivalent to 5000 mg/kg of compound). 5 males and 5 females were used. Symptoms and mortality after administration were recorded during an observation period of 14 days. At the end of the observation period surviving animals were killed by exsanguination under ether anaesthesia and an autopsy performed.

GLP compliance:

no

Remarks:

pre GLP

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Breeding: testing laboratory.
- Age at study initiation: 7 weeks.
- Weight at study initiation: average body weight of 163 g for females and 206 g for males.
- Housing: rats were caged singly.
- Fasting period before study: 18 hours
- Diet: commercial palletad diet (Dakes Special Diet with added Vit. E), ad libitum.
- Water: ad libitum.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Photoperiod: 12 hours artificial light and 12 hours darkness, in each 24 hour period.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25 % w/v solution of the compound in tap water.
- Rate: 20 ml/kg.

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females

Details on study design:

- Duration of observation period following administration: 14 days.
- Observations and weighing: deaths and clinical symptoms were recorded.
- Necropsy of survivors performed: 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:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the 14 day observation period.

Clinical signs:

other: Faeces were stained orange 24 hours after administration of the compound.

Gross pathology:

At autopsy no changes in organs or tissues caused by the administration of the test compound were seen.

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

LD50 > 5000 mg/Kg bw

Executive summary:

Method

Healthy Sprague-Dawley were administered with a 25 % w/v solution of the compound in tap water, as a single dose by gavage at a rate of 20 ml/kg (equivalent to 5000 mg/kg of compound). 5 males and 5 females were used. Symptoms and mortality after administration were recorded during an observation period of 14 days. At the end of the observation period surviving animals were killed by exsanguination under ether anaesthesia and an autopsy performed.

Results

Faeces were stained orange 24 hours after administration of the compound. No deaths occurred during the 14 day observation period. At autopsy no changes in organs or tissues caused by the administration of the test compound were seen.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Justification for type of information:

Justification for Read Across is detailed in the report attached to the IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Principles of method if other than guideline:

Substance was administered, as water solution, by gavage to 10 rats. A single dose of 5000 mg/kg was given. Animals were observed for the successive 14 days.

GLP compliance:

no

Remarks:

pre GLP

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 150 - 200 g

Route of administration:

oral: gavage

Vehicle:

water

Doses:

5000 mg/kg

No. of animals per sex per dose:

10 animals

Details on study design:

Duration of observation period following administration: 14

Statistics:

Probit-Analysis (Fink and Hund, Arzneimitteiforsch. 15, 1 965, 62).

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred.

Clinical signs:

other: Poisoning symptom: none

Interpretation of results:

other: not classified, according to the CLP Regulation

Conclusions:

LD50 > 5000 mg/Kg bw

Executive summary:

Method

Substance was administered, as water solution, by gavage to 10 rats. A single dose of 5000 mg/kg was given. Animals were observed for the successive 14 days. Unfortunately, only a summary sheet is available, thus details on testing procedures and results are lacking.

Results

No death occurred and no poisoning symptoms were recorded.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

According to the CLP Regulation (EC 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 greater than 5000 mg/kg bw.

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