Reaction mass of trisodium [m[7-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-8-hydroxynaphthalene-1,6-disulphonato(7-)]]dicuprate(3-) and trisodium [m[4-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-3-hydroxynaphthalene-2,7-disulphonato(7-)]]dicuprate(3-)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Direct Blue 251

IUPAC Name:

Direct Blue 251

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: animals were purchased from KFM, CH-4414 Füllinsdorf, Switzerland.
- Weight at study initiation: weight ranged between 160 - 190 g.
- Fasting period before study: access of food only was prevented approximately 18 hours prior and four hours after the dosing.
- Housing: rats were housed individually in Macrolon cages.
- Diet: standard laboratory pelleted diet (KLIBA no. 24-343-4 fro Klingentalmfihle AG., Basle), ad libitum. The batch of diet used for the study was analysed for chemical and microbiological contaminants.
- Water: ad libitum. The water bottles were with-drawn two hours prior and four hours after dosing.
- Acclimation period: about 5 days prior to the start of the study.

ENVIRONMENTAL CONDITIONS
- Temperature: the room temperature was maintained within a median of 23 ± 2 °C recorded by a maximum-minimum thermometer.
- Humidity: 30 - 70 %, measured by a hygrometer.
- Air changes: the rate of air exchange was maintained at approximately 15 changes/hour.
- Photoperiod: the light/dark cycle was 12 hours.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: the test item was diluted with WEM (33.3 %)

Details on oral exposure:

Dosed volumee: 15 ml per kg body weight.
The sample was prepared immediately before dosing.

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5 male and 5 female rats

Details on study design:

PRELIMINARY STUDY
A preliminary study was carried out to establish a dosing regimen, using groups of two males and two females at one dosage, using 15 ml/kg body weight.
- Duration of observation period following administration: 14 days

MAIN STUDY
- Duration of observation period following administration: 14 days
- Frequency of weighing: individual body weights on day 1, 7 and 14.
- Necropsy of survivors performed: surviving animals were killed after two weeks. All animals which died during the study and those killed after two weeks were subjected to a macroscopic postmortem examination. The macroscopic appearance of the abnormal organs was recorded.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occurred during both the preliminary and main experiments.

Clinical signs:

The animals were flaccid, weak and showed a rough coat.

Gross pathology:

No special findings.

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

LD50 (male and female) > 5000 mg/kg bw

Executive summary:

Young adult male and female rats were used to evaluated the oral acute toxicity potential of the test item. A group of 5 males and 5 females rats were dosed at 5000 mg/kg bw. The substance was administered at the dose volume of 15 ml/kg bw, by gavage. During the observation period of 14 days, all signs of toxicity were recorded. All rats were sacrificed terminally and examined macroscopically in an attempt to identify any target organs.

No deaths occurred during both the preliminary and main experiments; the animals were flaccid, weak and showed a rough coat. Gross examination did not revealed any special finding.

Conclusion

LD50 (male and female) > 5000 mg/kg bw