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Diss Factsheets

Administrative data

acute toxicity: oral
Type of information:
other: read across from similar substance
Adequacy of study:
key study
Study period:
From the 19th February to the 05th of March, 2003
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Study conducted according to internationally accepted testing guideline

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 3-[[ethyl[4-[[4-[(3-sulphonatophenyl)azo]-1-naphthyl]azo]phenyl]amino]methyl]benzenesulphonate
EC Number:
EC Name:
Disodium 3-[[ethyl[4-[[4-[(3-sulphonatophenyl)azo]-1-naphthyl]azo]phenyl]amino]methyl]benzenesulphonate
Cas Number:
Molecular formula:
Disodium 3-[[ethyl[4-[[4-[(3-sulphonatophenyl)azo]-1-naphthyl]azo]phenyl]amino]methyl]benzenesulphonate
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
Diluited in bidistilled water

Test animals

Details on test animals or test system and environmental conditions:
Source: Charles River (France), Charles River Laboratories Espana S.A.
Weight at study initiation: 85 to 99 g
Weight at study initiation (preliminary): 115 to 120 g
Weight during the study: 120-142 g (males), 108-123 g (females)
Age: four weeks
Date of arriving: 5, 12 Fenruary 2003
Identification:by code, ear-punch technique, before the administration
Group size:
preliminary study (500 mg/Kg to a female and afterwards to another female at 2000 mg/Kg)
main study (2000 mg/Kg, 3 to 7 males, 8 to 12 females)
Housing: Makrolon cages (59x38.5x20), with Ultrasorb sawdust bedding, supplied by PanLab, Spain. max 5 rats per cage.
Sawdust bedding replaced during the period of fasting
Diet: rat food -UAR A04C (France), analysed to detect contaminants, Batches 21118 and 21206, ad libitum. Food was withdrawn 16-18 hours before the administration
Water: ad libitum, available in bottles, analysed to detect contaminants (Spain). After 3-4 following the tratment, free access to food
Acclimatization: at least for 5 days, for veterany observation

Temperature: 21-25 °C
Humidity: 30-70 % (occasionally 75%)
Photoperiod (hrs dark / hrs light): 12 hours

Administration / exposure

Route of administration:
oral: gavage
Details on oral exposure:
Gastric gavage with metal cannula
One dose at volume 10 mL/kg, 500 mg/Kg, 2000 mg/Kg (Preliminary study)
One dose at volume 10 mL/kg, 2000 mg/Kg (Preliminary study)
The quantity of the test item calculated from the body weight

No. of animals per sex per dose:
10, 5 rats (5 male, 5 female)
Details on study design:
Preliminary study
Mortality in one animal
Signs of extreme toxicity in one animal
Twice a day for seven days

Main study
Mortality: immediately before administration and on days 7 and 14
Clinical signs: immediately before administration and on days 7 and 14 (changes in skin or furs, eyes and mucous membranes, respiratory, circulatory, central nervous and autonomic nervous systems, somatomotor activity and behaviour)
Body weight: dayly for three days
Necropsies: The animals were submitted to a gross necropsy at the end of the observation period. Inspection of intact animals and all superficial tissues, followed by the observation of the viscera of the cranial, thoracic and abdominals cavities.

Results and discussion

Preliminary study:
No mortalities occurred at 500 and 2000 mg/Kg bw (preliminary study)
No extreme toxicity occurred at 500 and 2000 mg/Kg bw (preliminary study)
Normal body weight at 500 and 2000 mg/Kg bw (preliminary study)

Sign of irritability 4-5 hours after the administration; dark coloured liquid faeces at 500 mg/Kg
Hunched back and ataxia from 4-5 hours after treatment; Ataxia and piloerection the first and the second day of the observation period, at 500 mg/Kg

Sign of irritability, hunched back and ataxia between 5-15 minutes and 5-6 hours after administration of 2000 mg/Kg.
Decreased motor activity and palpebral ptosis at 2-3 hours following treatment at 2000 mg/Kg
Ataxia on the first day of the observationanf third day, at 2000 mg/Kg

Effect levels
Dose descriptor:
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
No mortalities occurred at 2000 mg/Kg bw
Clinical signs:
Hunched back and ataxia were recorded after 90-120 minutes following the treatment in all the animals administered at the dose of 2000 mg/kg. These animals produced dark-coloured liquid faeces from 90-120 minutes. The dark colour of the faeces could be related to the colour of the test item. Irritability was observed in most of the animals treated at this dose between 90-120 minutes and 2-3 hours after the administration. All the animals administered at the dose of 2000 mg/kg had ataxia and, some of them, pasty faeces during the first day of the observation period.
Body weight:
Mean body-weight gain normal.
Gross pathology:
No macroscopic alterations were detected during the necropsies done on all the animals of the Main Study, except for one male administered at the dose of 2000 mg/kg which had a slightly dilated and congestive jejunum and one female treated at the same dose, in which a reddish area was observed on the intemal surface of the abdominal musculature.

Applicant's summary and conclusion

Interpretation of results:
other: not classified
Classification criteria according to the CLP Regulation 1272/2008 and its amendments
Preliminary Study: 500 mg/kg no mortality
Main study: 2000 mg/kg no mortality
Executive summary:


The acute oral toxicity was determined according to the EU Method B.1 (Acute Toxicity (Oral), in GLP.

In the Preliminary Study the test item was administered to one female at the dose of 500 mg/kg.


As no mortality was recorded in this animal, another female was administered at the dose of 2000 mg/kg, and this too survived the treatment.


In the Main Study ve males and ve females were treated at the dose of 2000 mg/kg and no mortality was recorded, hunched back, ataxia, irritability and liquid dark feaces were observed.