2-Naphthalenesulfonic acid, 4-hydroxy-3-[2-[2-methoxy-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-7-[(sulfomethyl)amino]-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, sodium salt (1:5)

Endpoint summary

Description of key information

The test substance is considered to be of low acute oral and dermal toxicity with LD50 values >2,000 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

Study period:

From Mar. 22, 2005 to Apr. 06, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Gartenstrasse 27. D-33178 Borchen
- Age at study initiation: 6-10 weeks
- Weight at study initiation: 178.5± 8.8 g (mean)
- Fasting period before study: yes
- Housing: In transparent macrolon® cages (type IV) on soft wood granulate in an air-conditioned room, 3 animals per cage
- Diet: ssniff® R/M-H (V 1534), ad libitum
- Water: tap water in plastic bottles, ad libitum

- Acclimation period: At least 5 d
- Animal identification: fur marking with KMnO4 and cage numbering

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C (short lasting deviations were permissible, e.g., during cleaning processes)
- Humidity: 50±20%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From Mar. 22, 2005 to Apr. 06, 2005

Route of administration:

oral: gavage

Vehicle:

other: deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20% suspension in deionized water
- Amount of vehicle (if gavage): 10 mL/kg bw

DOSAGE PREPARATION: Test substance was suspended in the stated concentration in deionized water and distributed homogeneously by means of a magnetic stirrer

CLASS METHOD: Acute toxic class method
- Rationale for the selection of the starting dose: The acute oral toxicity of test substance was tested only at a dose level of 2,000 mg/kg bw (limit test) according to toxicity data of related compounds. The animals received the test substance as a 20% suspension in deionized water, the administration volume was 10 mL/kg bw. As no compound-related mortality was produced in this limit test, no further dose was tested.

Doses:

2,000 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Twice every day (in the morning and in the afternoon), on weekends and public holidays only once
- Necropsy of survivors performed: yes
- Other examinations performed: Mortality, body weight, clinical observations and macroscopic examinations

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the whole study.

Clinical signs:

other: Dark red discolored feces was the only clinical sign observed in the animals at the time point of 4-8 h after the administration of the test substance: From Day 2 until the end of the study no symptoms were observed.

Gross pathology:

The animals killed at the end of the observation period showed no macroscopically visible changes.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the oral LD50 value for the test substance was found to be >2000 mg/kg bw.

Executive summary:

A study was conducted to assess the acute toxicity of the test substance on rats according to EU Method B.1 and OECD Guideline 423, in compliance with GLP.

The substance was tested only at a dose of 2,000 mg/kg bw (limit test). The substance was administered by gavage to fasted animals as a 20% suspension in deionized water. The administration volume was 10 mL/kg bw. The observation period following treatment lasted for 14 d. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time, the animals were weighed weekly. At the end of the observation period, the animals were sacrificed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes.

No mortality or adverse effect on development of body weight was observed. No clinical signs, except discoloured feces at the time point of 4-8 h after administration, were observed. The macroscopic examination after sacrifice did not reveal any adverse effects.

Hence, under the test conditions, the oral LD50 was > 2,000 mg/kg bw (Kauffmann, 2005a).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

High quality study.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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 Mar. 30, 2005 to Apr. 13, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

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
- Source: Harlan Winkelmann GmbH, Gartenstrasse 27, D-33178, Borchen.
- Age at study initiation: 6-10 weeks
- Weight at study initiation: 248.8±15.4 g (males); 215.2±5.6 g (females)
- Housing: in transparent macrolon® cages (type III) on soft wood granulate* in an air-conditioned room, 1 animal per cage
- Diet: ssniff® R/M-H (V 1534), ad libitum
- Water: tap water in plastic bottles, ad libitum
- Acclimation period: At least 5 d

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 50±20 %
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From Mar. 30, 2005 to Apr. 13, 2005

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: 30cm2
- Type of wrap if used: Fixomull and Elastoplast

REMOVAL OF TEST SUBSTANCE
- Washing: With warm water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied: 0.5 g
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied: 0.3 mL

Duration of exposure:

24 h

Doses:

2,000 mg/kg bw

No. of animals per sex per dose:

5/sex

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Twice every day (in the morning and in the afternoon), on weekends and public holidays only once.
- Necropsy of survivors performed: yes
- Other examinations performed: Mortality, body weight, clinical observations and macroscopical examinations

- Overall study design: Before dermal treatment the fur was mechanically removed from the dorsal skin of the animals over an area of approximately 30 cm². 0.5 g test substance was moistened with 0.3 mL deionized water on a two-ply gauze and an aluminum foil (6x8 cm) and distributed as uniformly as possible. Together with the foil the test substance was administered to the shaved and intact dorsal skin. The foil was held in place with an elastic plaster bandage fixed around the animal's body (Fixomull and Elastoplast, 8 cm in width, both manufactured by Beiersdorf Aktiengesellschaft).
At the end of the dermal exposure period of 24 h the bandage was removed and the treated skin area washed with warm water in order to remove any unabsorbed remnants of the test substance.
The observation period after the dermal administration lasted for 14 d. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time the animals were weighed weekly. At the end of the observation period the animals were sacrificed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the whole study

Clinical signs:

other: No test substance related symptoms were observed after administration of 2,000 mg/kg bw. The skin of the animals showed substance related discolorations (by compound) starting on Day 2 in all animals and lasting up to Day 13 of the study on some animals.

Gross pathology:

The animals killed at the end of the observation period showed no macroscopically visible changes.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the dermal LD50 value for the test substance was found to be >2000 mg/kg bw.

Executive summary:

A study was conducted to assess the acute toxicity of the test substance on rats according to EU Method B.3 and OECD Guideline 402, in compliance with GLP.

The animals (5/sex) were exposed to the test substance at the dose rate of 2,000 mg/kg bw for the period of 24 h. At the end of the dermal exposure period, the bandage was removed and the treated skin area washed with warm water in order to remove any unabsorbed remnants of the test substance. The observation period after the dermal administration lasted for 14 d. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time the animals were weighed weekly. At the end of the observation period the animals were sacrificed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes.

No deaths occurred during the whole study. One female animal showed a slight loss of body weight at the end of the study. In the other female and male animals development of body weight was not impaired. No test substance related symptoms were observed after administration of 2,000 mg/kg bw. The skin of the animals showed substance related discolorations (by compound) starting on Day 2 in all animals and lasting up to Day 13 of the study on some animals.

Hence, under the test conditions, the dermal LD50 value for the test substance was found to be >2,000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

High quality study

Additional information

Oral route:

A study was conducted to assess the acute toxicity of the test substance on rats according to EU Method B.1 and OECD Guideline 423, in compliance with GLP. The substance was tested only at a dose of 2,000 mg/kg bw (limit test). The substance was administered by gavage to fasted animals as a 20% suspension in deionized water. The administration volume was 10 mL/kg bw. The observation period following treatment lasted for 14 d. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time, the animals were weighed weekly. At the end of the observation period, the animals were sacrificed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes. No mortality or adverse effect on development of body weight was observed. No clinical signs, except discoloured feces at the time point of 4-8 h after administration, were observed. The macroscopic examination after sacrifice did not reveal any adverse effects. Hence, under the test conditions, the oral LD50 was > 2,000 mg/kg bw (Kauffmann, 2005b).

Dermal route:

A study was conducted to assess the acute toxicity of the test substance on rats according to EU Method B.3 and OECD Guideline 402, in compliance with GLP. The animals (5/sex) were exposed to the test substance at the dose rate of 2,000 mg/kg bw for the period of 24 h. At the end of the dermal exposure period, the bandage was removed and the treated skin area washed with warm water in order to remove any unabsorbed remnants of the test substance. The observation period after the dermal administration lasted for 14 d. Symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once. During this time the animals were weighed weekly. At the end of the observation period the animals were sacrificed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes. No deaths occurred during the whole study. One female animal showed a slight loss of body weight at the end of the study. In the other female and male animals development of body weight was not impaired. No test substance related symptoms were observed after administration of 2,000 mg/kg bw. The skin of the animals showed substance related discolorations (by compound) starting on Day 2 in all animals and lasting up to Day 13 of the study on some animals. Hence, under the test conditions, the dermal LD50 value for the test substance was found to be >2,000 mg/kg bw (Kauffmann, 2005c).

Justification for classification or non-classification

Oral route:

Based on the results of an acute oral toxicity study, the test substance does not need to be classified for this endpoint according to the EU CLP criteria (EC 1272/2008) as well as EU Directive 67/548/EEC.

Dermal route:

Based on the results of an acute dermal toxicity study, the test substance does not need to be classified for this endpoint according to the EU CLP criteria (EC 1272/2008) as well as EU Directive 67/548/EEC.