Benzenesulfonic acid, 2,2'-(1,2-ethenediyl)bis[5-nitro-, disodium salt, reaction products with 4-[(4-aminophenyl)azo]benzenesulfonic acid monosodium salt

Endpoint summary

Description of key information

OECD 401, oral, rat, limIt dose LD50 > 2000 mg/kg.

Oral,rat fed with different doses, limit dose LD50 = 8700 mg/kg.

EU Method B.1, oral, rat, limit dose LD50 > 2000 mg/kg

 

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:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Source study has reliability 2. Details on the read across are attached in section 13.

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

not specified

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: young and healthy rats
- Weight at study initiation: 208 to 231 g
- Housing: Makrolon (48 × 27 × 20 cm) Tecniplast brand
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Humidity: 55 ± 25 %
- Air changes per hr: 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The sample is diluted 1/10 with water
MAXIMUM DOSE VOLUME APPLIED: 1.69 ml per 100 g of body weight, equivalent to 2000 mg/kg bw

Doses:

2000 mg/kg

No. of animals per sex per dose:

3 males and 3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: every day
- Necropsy of survivors performed: yes
- Other examinations performed: skin, hair, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavioral patterns.
- Special attention to: tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Not observed

Clinical signs:

other: All behaviours were considered normal. No toxicity signs were attributed to test substance.

Body weight:

other body weight observations

Remarks:

The body weight variation registered is considered normal.

Gross pathology:

No significant macroscopic changes in any of the animals.

rat	weight in g
	initial	day 7	final
M	215	280	309
M	231	297	320
M	208	284	313
F	164	198	207
F	170	195	214
F	171	198	215

Interpretation of results:

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

Conclusions:

The substance was tested according to guideline EU Method B.1.
LD50 in rats was found to be > 2000 mg/kg.

Executive summary:

Method

Acute oral toxicity was assessed in a limit test. A dose of 2000 mg/kg bw was administered in several phases in order to assess mortality in 6 rats (3 males and 3 females).

An individual weight control and general examinations during the observation period of 14 days were performed.

Results

No clinical signs were noted and no mortality occurred. Therefore, LD50 > 2000 mg/kg.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Source study has reliability 2. Details on the read across are attached in section 13.

Principles of method if other than guideline:

Rats were feeded with different doses of substance.
Symptoms and mortality were recorded during an observation period of 15 days.
The LD50 was calculated by probit analysis method (Maximum likelyhood, Goulden A., Methods of Statistical Analysis, John Wiley and Sons, 1960, 3rd printing, pages 404-408).

GLP compliance:

not specified

Test type:

acute toxic class 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
- Weight at study initiation: 100 to 127 g
- Fasting period before study: overnight
- Housing: in groups of 5
- Diet: standard diet of Nafag ad libitum
- Water: fresh water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1°C
- Humidity: 55 ± 5 %
- Photoperiod: 14 hours light day

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

2%

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25 %
- Amount of vehicle (if gavage): 2 %

Doses:

1000, 3000, 6000, 8000, 10000 mg/kg

No. of animals per sex per dose:

5 male/5 female per dose as described above

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations: symptoms and mortality were recorded daily for 15 days.
-Frequency of weighing: the animals were weighed during the pre-test and on day15 of test.
- Necropsy of survivors performed: no

Statistics:

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

Sex:

male/female

Dose descriptor:

LD50

Effect level:

8 700 mg/kg bw

Based on:

test mat.

95% CL:

> 8 110 - < 9 340

Mortality:

No mortality occurred.

Clinical signs:

other: ditto, venterocumbency, hypoventilation, ataxia, transient diarrhea was seen at doses of 6000, 8000,10000 mg/kg.

Gross pathology:

Not recorded. Necropsy has not been done.

 

dose	vehicle	conc. %	mortality				bodyweight (g)		symptoms
			absolute		%	day of test	pre-test	on day 15 of test
			F	M
1000	CMC 2 %	25	0/5	0/5	0	-	100	182	none
3000		25	0/5	0/5	0	-	100	175	reduction in spontaneous motility lasting > 3 h. After 24 h, no symptoms
6000		30	0/5	0/5	0	-	127	215	ditto, hypoventilation, ataxia, transient diarrhea
8000		30	0/5	2/5	20	1	100	170	ditto, hypoventilation, ataxia, transient diarrhea
10000		25	5/5	4/5	90	1	100	147	ditto, venterocumbeny

Interpretation of results:

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

Conclusions:

Rats were feeded with different doses of substance.
The acute oral LD50 in rats was calculated to be 8700 mg/kg.

Executive summary:

Method

After 5 days of acclimatization, groups of 5 male and 5 female rats were given various single doses of test substance (1000, 3000, 6000, 8000, 10000 mg/kg) suspended in carboxymethylcellulose 2 % (CMC). Symptoms and mortality after administration were recorded during an observation period of 15 days.

Results

No mortality was recorded up to a single dose of 6000 mg/kg. At doses of 8000 and 10000 mg/kg, 20 % and 90 % mortality was recorded. The acute oral LD50 in rats was calculated to be 8700 mg/kg. Dose-dependent clinical signs were noted, e.g. decreased motility, hypoventilation, ataxia, diarrhea, venterocumbeny.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Source study has reliability 2. Details on the read across are attached in section13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Qualifier:

according to guideline

Guideline:

other: WHO/VBC 88.953

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: TIf: RAI f (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CIBA-GEIGY Limited Animal Production 4332 Stein / Switzerland
- Weight at study initiation: 177 to 206 g
- Fasting period before study: overnight
- Housing: in Macrolon cages type 4, with standardized soft wood beeding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Air changes: 15 per h
- Photoperiod: 12 h /day light cycle.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Single oral dose, by gastric intubation (gavage)
Vehicle: distilled water
Amount of vehicle: 10 ml/kg bw

Doses:

2000 mg/kg (males and females)
Volume applied: 10 ml/kg bw

No. of animals per sex per dose:

5 males and 5 females (total number of animals: 10)

Control animals:

yes

Details on study design:

Observations and records
Mortality: daily; a.m. and p.m. on working days, a.m. on weekend days.
Signs and symptoms: daily for 14 days.
Body weight: immediately before administration and on days 7 and 14.
Necropsies: animals submitted to a gross necropsy at the end of the observation period.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities occurred.

Clinical signs:

other: Piloerection, hunched posture, and dyspnea were seen, being common symptoms in acute tests. Additionally, diarrhea was observed in females. Animals recovered within 5 days.

Gross pathology:

At necropsy, a spotted thymus was found in one female. No deviations from normal morphology were found in the remaining animals.

	dose (mg/kg)	number of animals	number of deaths
males	2000	5	0
famales	2000	5	0

 

Interpretation of results:

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

Conclusions:

The substance was tested according to the guidelines OECD 401.
Upon an acute oral administration and a 14 day post-treatment observation period in rats of both sexes, no mortality was recorded. Therefore, in male and female rats:
LD0 = 2000 mg/kg
LD50 > 2000 mg/kg

Executive summary:

Method

Acute oral toxicity to rats was assessed in a limit test. A single dose of 2000 mg/kg was given by gavage to 10 young adult rats (5 males and 5 females Tif: Rai f). Rats were obserevd daily for 14 days after dosing. Mortality, signs and symptoms were recorded daily; body weight was measured immediately before administration and on days 7 and 14; necropsy was done at the end of the observation period.

Results

No mortality was seen during the 14 -days observation period, thus LD0 = 2000 mg/kg and LD50 > 2000 mg/kg.

Clinical signs as piloerection, dyspnea, hunched posture and diarrhea were recorded.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

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

No data on target substance was available, thus a read across approach was followed as detailed in section 13. Experimental data on Similar Substance 01 and Similar Substance 02 was used.

A study on Similar Substance 01 was selected as key study, mainly due to the high purity of test sample (91 % of active substance). It was a limit test according to OECD guideline 401. Rats were dosed by gavage 2000 mg/kg bw. No deaths occurred and no remarkable clinical signs and macroscopic changes were noted in both male/female. Accordingly, LD50 > 2000 mg/kg.

In an older test on Similar Substance 01 (66 % of active substance), rats were dosed with 1000 to 10000 mg/kg of test substance and a LD50 of 8700 mg/kg was obtained by probit analysis.

In a study conducted in rats on Similar Substance 02 (10 % of active substance) according to EU method B.1, a LD50 > 2500 mg/kg was found.

 

 

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 oral, dermal or inhalation route according to numeric criteria. Acute toxicity values are expressed as LD50 (oral, dermal) or LC50 (inhalation) values as well as acute toxicity estimates (ATE).

 

In available studies in rats, oral LD50 values above 2000 mg/kg bw were found. Accordingly, target substance was not classified for acute oral toxicity in the CLP Regulation (EC 1272/2008).