Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo)-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfonyl)benzenesulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From July 27, 2005 to August 12, 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

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:

-

EC Number:

475-300-1

EC Name:

-

Cas Number:

910030-59-0

Molecular formula:

Hill formula: C29H25N8Na5O22S7 CAS formula: C29H30N8O22S7.5Na

IUPAC Name:

Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo )-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfony l)benzenesulfonate

Test material form:

solid: particulate/powder

Details on test material:

see below

Test animals

Species:

rat

Strain:

Wistar

Remarks:

CRL:(WI) BR rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (Europe) laboratories INC.
- Hygienic level at arrival: SPF
- Age at study initiation: Young healthy adult rats, less than 10 weeks old
- Body weight range at treatment: 175-188g
- Housing: II type polypropylene/polycarbonate, 3 animals per cage
- Bedding: Laboratory bedding
- Diet: Ssniff RIM-Z+H, ad libitum
- Water: Tap water from 500 mL bottle, ad libitum
- Acclimation period: 14 d
-Animal identification: Numbers on the tail written with a marker pen
ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30 - 70%
- Air changes: 8-12 air exchanges/hour by central air-condition system
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: July 27, 2005 to August 12, 2005

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 1 % aqueous methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Lot/batch no.: 05-07/01
- Expiry date: August 4, 2005
- Storage: In refrigerator

MAXIMUM DOSE VOLUME APPLIED: 1 mL/100 g bw (i.e., 10 mL/kg bw)

DOSAGE PREPARATION: For treatment the test substance was applied in a concentration of 200 mg/mL in 1 % aqueous methylcellulose. Formulations were prepared just before the treatment.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No mortality was expected at 2,000 mg/kg bw dose level, so this was chosen as the starting dose.

Doses:

2,000 mg/kg bw

No. of animals per sex per dose:

6 animals (i.e., 3 animals/group)

Details on study design:

Test procedure
A single oral administration - followed by a 14 d observation period was performed by gavage. On the day before each treatment the animals were fasted. The food but not water was withheld during an overnight period. Animals were weighed just before the application. The test substance was administered by oral gavage in the morning hours. The food was given back 3 h after the treatment. A constant treatment volume of 1 mL/100 g bw was applied.

Three female animals were treated with a dose level of 2,000 mg/kg bw of test substance in the first step. All animals survived; so three further animals were dosed at 2,000 mg/kg bw dose level next day, as the second step. No mortality occurred in either step, so the test was finished meeting the stopping criteria ofguidelines.

Clinical Observations
Careful clinical observation was made once within the first 30 minutes, then 30 minutes, 1 h, 2 h, 3 h, 4 h, 6 h after the treatment and once each day for 14 d thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern as well. Particular attention was directed to the observation of tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma.

Body Weight Measurement

The body weight was recorded on Day 0 (shortly before the treatment), then on Days 7 and 14 with precision of 1g.

NECROPSY

All animals were subjected to gross pathology. Animals were exsanguinated under pentobarbital anesthesia. After examination of the external appearance the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. Abnormalities were recorded on post mortem data sheets.

Results and discussion

Effect levels

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:

Test substance caused no mortality in female CRL:(Wl) BR rats after a single oral (i.e., by gavage) administration of 2,000 mg/kg bw.

Clinical signs:

other: Slight diarrhoea was observed in two animals from 2 h after the dosing to the end of the day. No clinical symptoms appeared on the day of the treatment on the other animals and the following 14 d observation period. The physical condition and behaviour of

Gross pathology:

No macroscopic alterations related to the toxic effect of test substance were found at necropsy. The pinprick-sized haemorrhages (3/6) observed in the lungs were attributed to the termination procedures and exsanguination.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

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

Executive summary:

A study was conducted to assess the acute oral toxicity of the test substance in CRL:(WI) BR female rats according to OECD Guideline 423, EPA OPPTS 870.1100 and EU Method B.1 tris, in compliance with GLP.

Group of three female rats received a single oral (gavage) dose of 2,000 mg/kg bw in a first and second step, respectively. A concentration of 200 mg/mL of test substance was prepared in 1 % aqueous rnethylcellulose and administered at a volume of 1 mL/100 g bw (i.e., 10 mL/kg bw).

 

No mortality occurred, no effect on body weight was observed and no significant macroscopic abnormalities were seen at necropsy. Except for slight diarrhoea in two animals from 2 h after the dosing to the end of the day, no clinical symptoms were observed in other animals.

 

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