Benzoic acid, 2-hydroxy-5-[[4-[[4-[[8-hydroxy-7-[[4-[(8-hydroxy-3,6-disulfo-1-naphthalenyl)azo]-2-methoxy-5-methylphenyl]azo]-3,6-disulfo-1-naphthalenyl]amino]-6-(phenylamino)-1,3,5-triazin-2-yl]amino]phenyl]azo]-, pentasodium salt

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:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Principles of method if other than guideline:

Acute oral toxicity in rats. Groups of 5 rat/sex were dosed at 2500, 3000, 3500, 4200 mg/kg by gavage and observed for 14 days after dosing.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Direct Green 026

IUPAC Name:

Direct Green 026

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5 weeks
- Weight at study initiation: 137 g (M), 123 g (F)
- Fasting period before study: 18 hours
- Housing: singly
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Photoperiod: 12 light and 12 darkness

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 25, 30, 35, 42 %
- Amount of vehicle: 10 ml/kg

Doses:

2500, 3000, 3500, 4200 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs

Statistics:

Method of Litchfield J.T. and Wilcoxon F.W. (1949), J. Pharmac. Expt. Therap.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3 800 mg/kg bw

Based on:

test mat.

95% CL:

> 3 300 - < 4 300

Clinical signs:

A few rats at each dose level showed hyperactivity and/or convulsions when disturbed.

Other findings:

Autopsy reports on animals which died during the study:
3000 mg/kg 1 M: dead 47 hours after dosing. Green discolouration of the medulla of the kidneys.
3500 mg/kg 1 M: dead 13 hours after dosing. Green discolouration of the diaphragm and the medulla of the kidneys.
4200 mg/kg 1 M: dead 24 hours after dosing. Excess clear thoracic fluid.
1 F: dead 42 hours after dosing. Green discolouration of the diaphragm and the medulla of the kidneys.

Autopsy after 14 days observation: at 3000, 3500 and 4200 mg/kg, haemorrhages on lungs.

Any other information on results incl. tables

Dose and mortality data

dose	animals	mortality ratio		combined deaths	%
		M	F
2500 mg/kg	5/sex	0/5	0/5	0/10	0
3000 mg/kg	5/sex	2/5	1/5	3/10	30
3500 mg/kg	5/sex	3/5	2/5	5/10	50
4200 mg/kg	5/sex	2/5	2/5	4/10	40

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

LD50 = 3800 mg/kg in rats

Executive summary:

Method

Rats of both sexes dosed by gavage with 2500, 3000, 3500 and 4200 mg/kg as single oral dose. Observations were continued up to 14 days after dosing. Mortality and clinical signs were recorded. At the end of the observation period, surviving rats were killed and autopsy was performed.

Results

LD50 = 3800 mg/kg.

As for animals died during the study, green discouloration of diaphragm and medulla of kidneys were noted.

At autopsy after 14 days, hemorrhages on lungs were noted.