tetrasodium 3-amino-4-{[4-({4-fluoro-6-[phenyl(2-{[2-(sulfonatooxy)ethyl]sulfonyl}ethyl)amino]-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]diazenyl}-5-hydroxynaphthalene-2,7-disulfonate

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 September 27, 2004 to October 14, 2004

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 to 10 weeks
- Body weight range at treatment: 178-191 g
- Housing: In transparent macrolon cages (type IV) on soft wood granulate in an air-conditioned room, 3 animals per cage
- Diet: Ssnif 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
- Randomization procedure: Computer generated algorithm (archived with raw data) Randomization scheme 2004.0213

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

IN-LIFE DATES: From: To: September 27, 2004 to October 14, 2004

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionized

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20% solution in deionized water

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual): Test substance was dissolved in the stated concentration in deionized water and distributed homogeneously by means of a magnetic stirrer. The stability and the homogeneity of the test substance in the vehicle was determined by analytical methods.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Limit test (according to toxicity data of related compounds).

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

six (female only)

Control animals:

no

Details on study design:

TEST PROCEDURE
The prepared test substance was administered by gavage to fasted animals at the stated dosage. The observation period following treatment lasted for 14 d. Symptoms were recorded twice every day (i.e., 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 killed by carbon dioxide asphyxiation, dissected and examined for macroscopically visible changes.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occurred during the whole study.

Clinical signs:

other: Reddish discolored urine and feces were observed in the animals starting 2-4 h after the administration of test substance. From Day 3 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 study conditions, the oral LD50 was found to be >2000 mg/kg bw in rats.

Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance in female Hsd:Sprague Dawley (SD) rats according to OECD Guideline 423 and EU Method B.1, in compliance with GLP.

Group of six female fasted rats received a single oral (gavage) dose of 2000 mg/kg bw. A 20% solution of test substance was prepared in deionized water and administered at a volume of 10 mL/kg bw.

 

No mortality occurred, no effect on body weight was observed and no significant macroscopic abnormalities were seen at necropsy. Reddish discolored urine and feces were observed after the administration of test substance for up to 2 d. There were no further symptoms from Day 2 until end of the study.

 

Under the study conditions, the oral LD50 was found to be >2,000 mg/kg bw in rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

High quality study report

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 Sept. 30, 2004 to Oct. 14, 2004

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

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 to 10 weeks
- Weight at study initiation: 254 g (males) and 224 g (females)
- Housing: in transparent macrolon® cages (type III) on soft wood granulate* in an air-conditioned room, 1 animal per cage
- Diet (e.g. ad libitum): ssniff® R/M-H (V 1534), ad libitum
- Water (e.g. ad libitum): tap water in plastic bottles, ad libitum
- Acclimation period: at least five days

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

IN-LIFE DATES: From: Sept. 30, 2004, To: Oct 14, 2014

Type of coverage:

occlusive

Vehicle:

water

Remarks:

deionized

Details on dermal exposure:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with warm water in order to remove any unabsorbed remnants of the test substance.
- Time after start of exposure: 24 h

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

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

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occurred during the whole study

Clinical signs:

other: No symptoms were observed after administration of 2,000 mg/kg bw. The skin of the animals showed substance related discolorations (reddish). From Day 8 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 test substance was determined to have the median lethal dermal dose value (LD50) of greater than 2000 mg/kg bw.

Executive summary:

An acute dermal toxicity for the test subsatnce was conducted according to OECD guideline 402, EU method B.3 and EPA OPPTS 870.1200 and in compliance with GLP.

After administration of 2000 mg/kg bw dermally, the animals were observed for 14 d for clinical signs, mortality and body weight change. No clinical signs and mortality occurred. The skin of the animals showed substance related discolorations (reddish) from Day 2 up to Day 7 of the study. Two female animals showed loss of body weight during the first week of the study, which returned to normal until the end of the study. One female animal showed a loss of body weight at the end of the study. In the other animals development of body weight was not impaired. Development of body weight in male animals was not impaired. All animals were sacrificed at the end of the observation period. They showed no macroscopically visible changes.

Under the test conditions, the test substance was determined to have the median lethal dermal dose value (LD50) of greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

High quality study report

Additional information

Acute oral toxicity:

A study was conducted to assess the acute oral toxicity of the test substance in female Hsd: Sprague Dawley (SD) rats according to OECD Guideline 423 and EU Method B.1, in compliance with GLP. Group of six female fasted rats received a single oral (gavage) dose of 2,000 mg/kg bw. A 20% solution of test substance was prepared in deionized water and administered at a volume of 10 mL/kg bw. No mortality occurred, no effect on body weight was observed and no significant macroscopic abnormalities were seen at necropsy. Reddish discolored urine and feces were observed after the administration of test substance for up to 2 d. There were no further symptoms from Day 2 until end of the study. Under the study conditions, the oral LD50 was found to be >2,000 mg/kg bw in rats (Kaufmann, 2004a).

Acute dermal toxicity:

An acute dermal toxicity for the test substance was conducted according to OECD guideline 402, EU method B.3 and EPA OPPTS 870.1200 and in compliance with GLP. After administration of 2,000 mg/kg bw dermally, the animals were observed for 14 d for clinical signs, mortality and body weight change. No clinical signs and mortality occurred. The skin of the animals showed substance related discolorations (reddish) from Day 2 up to Day 7 of the study. Two female animals showed loss of body weight during the first week of the study, which returned to normal until the end of the study. One female animal showed a loss of body weight at the end of the study. In the other animals development of body weight was not impaired. Development of body weight in male animals was not impaired. All animals were sacrificed at the end of the observation period. They showed no macroscopically visible changes. Under the test conditions, the test substance was determined to have the median lethal dermal dose value (LD50) of greater than 2,000 mg/kg bw (Kaufmann, 2004b).

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).

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).