Benzenesulfonic acid, 2,2'-(1,2-ethenediyl)bis[5-[[4-(diethylamino)-6-[(4-sulfophenyl)amino]-1,3,5-triazin-2-yl]amino]-, sodium salt (1:4)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From September 29 to Ocober 15, 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 17 December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tetrasodium 2,2'-ethene-1,2-diylbis[5-({4-[diethylamino]-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)benzenesulfonate]

Cas Number:

42355-78-2

Molecular formula:

C40H40N12Na4O12S4

IUPAC Name:

tetrasodium 2,2'-ethene-1,2-diylbis[5-({4-[diethylamino]-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)benzenesulfonate]

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Únětice, Czech Republic
- Age at study initiation: 8 - 10 weeks at the time application
- Fasting period before study: twenty hours before oral administration the animals were not fed.
- Housing: animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage Velaz T4.
- Bedding: sterilized shavings of soft wood.
- Diet: ad libitum, ST - 1, VELAS a.s., Hrabanov 535, 289 22 Lysá nad Labem, CZ 801080-01.
- Water: ad libitum, drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law).
- Acclimation period: 20 days
- Health condition: certificate of good health condition from breeding farm; no signs of diseases were observed at clinical check-in, during the acclimatisation period and before the start of study.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 – 70 %
- Photoperiod: light period 12-hour light/12 hour dark.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua pro injection

Details on oral exposure:

Preparation and application of the test substance: immediately before application the test substance was weighed and mixed in vehicle. The test substance was administered dissolved in water pro injection.

Dosing: the test substance was administered to the stomach by tube. The single volume of administered solution was 1 ml/100 g of animal body weight.

Doses:

2000 mg/kg of body weight

No. of animals per sex per dose:

6 females (3 per group)

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: immediately before application the animals were weighed, 8th day and at the 15th day, before euthanasia of animals.
- Necropsy of survivors performed: yes.
- Clinical examination: after application the animals were observed individually. The first day twice (30 minutes and 3 hours after application), the second day twice (in the morning and in the afternoon) and daily thereafter for 14 days. Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.
- Pathological examination: all test animals survived to the end of study were sacrificed on the 15th day by injection of veterinary preparation T61 (1 ml iv.) and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No death occured.

Clinical signs:

other: No clinical signs of intoxication were observed during the study.

Gross pathology:

No pathologic changes were recorded.

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met

Conclusions:

The LD50 of the test substance for female rats is higher than 2000 mg/kg of body weight.

Executive summary:

Method

The aim of the study was to investigate acute toxic effects of the test substance after a single oral administration to Wistar CRL rats.

The testing was performed according Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008. The test substance was administered dissolved in water pro injection as a single dose, given orally via gavage to three groups of three female Wistar CRL rats. 

The dosing was performed sequentially in two groups of three females:

group No. 1 - first step using the starting dose of 2000 mg/kg of body weight

group No. 2 - second step using the same dose 2000 mg/kg.

Results

The test substance administered at the dose of 2000 mg/kg caused no death of animals. No clinical signs of intoxication were detected during whole study. No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test substance for female rats is higher than 2000 mg/kg of body weight.