A mixture of: trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-(4-nitro-2-sulfonatoanilino)phenylazo)phenolato)ferrate(1-); trisodium bis(2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-nitro-2-sulfonatophenylazo)phenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(3-sulfonatophenylazo)phenolato)ferrate(1-); disodium 3,3'-(2,4-dihydroxy-1,3(or 1,5 or 3,5)-phenylenediazo)dibenzenesulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From September 19, 1990 to October 18, 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1991

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-(4-nitro-2-sulfonatoanilino)phenylazo)phenolato)ferrate(1-); trisodium bis(2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-nitro-2-sulfonatophenylazo)phenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(3-sulfonatophenylazo)phenolato)ferrate(1-); disodium 3,3'-(2,4-dihydroxy-1,3(or 1,5 or 3,5)-phenylenediazo)dibenzenesulfonate

EC Number:

406-870-1

EC Name:

A mixture of: trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-(4-nitro-2-sulfonatoanilino)phenylazo)phenolato)ferrate(1-); trisodium bis(2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(4-nitro-2-sulfonatophenylazo)phenolato)ferrate(1-); trisodium (2,4(or 2,6 or 4,6)-bis(3,5-dinitro-2-oxidophenylazo)-5-hydroxyphenolato)(2(or 4 or 6)-(3,5-dinitro-2-oxidophenylazo)-5-hydroxy-4(or 2 or 6)-(3-sulfonatophenylazo)phenolato)ferrate(1-); disodium 3,3'-(2,4-dihydroxy-1,3(or 1,5 or 3,5)-phenylenediazo)dibenzenesulfonate

Cas Number:

115100-55-5

Molecular formula:

not applicable

IUPAC Name:

triiron(3+) nonasodium 2-(2-{4-hydroxy-2-oxido-5-[2-(3-sulfophenyl)diazen-1-yl]phenyl}diazen-1-yl)-4,6-dinitrobenzen-1-olate 2-(2-{5-[2-(3,5-dinitro-2-oxidophenyl)diazen-1-yl]-2-hydroxy-4-oxidophenyl}diazen-1-yl)-4,6-dinitrobenzen-1-olate tris(2-(2-{5-[2-(3,5-dinitro-2-oxidophenyl)diazen-1-yl]-4-hydroxy-2-oxidophenyl}diazen-1-yl)-4,6-dinitrobenzen-1-olate) 2-{2-[4-hydroxy-5-(2-{4-[(4-nitro-2-sulfophenyl)amino]phenyl}diazen-1-yl)-2-oxidophenyl]diazen-1-yl}-4,6-dinitrobenzen-1-olate

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
-Source: BRL Ltd., Basel, Switzerland
-Females (if applicable) nulliparous and non-pregnant: [yes/no]
-Age at study initiation: Approx. 8-9 weeks
-Weight at study initiation:
males; 179 - 203 g
females: 154 - 201 g
-Fasting period before study: Feed was withheld overnight prior to dosing until approxlmately 3-4 hours after administration of the test substance.
-Housing: Group housed, five per sex to a cage, using polycarbonate cages containing purified sawdust as bedding material (Woody SPF, supplied by Broekman Instltute, Someren, The Netherlands).
-Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (Kliba 343 from Klingentalmühle AG, Kaiseraugst, Switzerland).
-Water (e.g. ad libitum): Free access to tap-water.
-Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 21 ± 3 °C
-Humidity (%): 40-70 %
-Air changes (per hr): 15 air changes per hour
-Photoperiod (hrs dark / hrs light): Lighting was 12 hours artificial fluorescent light and 12 hours dark per day.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
-Amount of vehicle (if gavage): 20 ml/kg body weight (Group 1 ), 10 ml/kg body weight (Group 2 and 3 )

Doses:

Group 1: 5000 mg/kg body weight.
Group 2: 2800 mg/kg body weight.
Group 3: 1580 mg/kg body weight

No. of animals per sex per dose:

5 per sex per doses

Control animals:

no

Details on study design:

-Duration of observation period following administration: 14 days
-Frequency of observations Mortality / Viability: At periodic intervals on the day of dosing (day 1) and twice dally thereafter for 14 days
-Frequency of observations and weighing: Test days 1 (pre-administration), 8 and 15.
-Necropsy of survivors performed: yes
All animals killed for humane reasons or animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide asphyxiation and subjected to necropsy. All animals assigned to the study were subjected to necropsy.
The necropsies were performed by experienced prosectors and descriptions of all macroscopic abnormalities recorded.
-Other examinations performed: At periodic intervals on the day of dosing (day 1) and once daily thereafter for 14 days.
All signs of reaction to treatment were recorded with particular attention paid to changes in the skin, fur, eyes and mucous membranes, as well as to behaviour pattern, tremors, convulsion, salivation, diarrhoea, lethargy, sleep and coma.

Statistics:

The LD50 value for females alone and the slope of the dose mortality curve were estimated using the Maximum likelihood method (Finney, O.G. Probit Analyses, 3rd Edn., London, Cambridge, University Press, 1971).

Results and discussion

Effect levels

Effect levels 1

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

LD50

Effect level:

4 047 mg/kg bw

Based on:

test mat.

Mortality:

4 females rats are dead within 24 hours of dosing.

Clinical signs:

Signs of systemic toxicity observed during the study period in each dose group were as follows:
5000 mg/kg body weight: Piloerection, lethargy, hunched posture, ataxia, alopecia
2800 mg/kg body weight: Piloerection, lethargy, alopecia
1580 mg/kg body weight: No abnormities
All surviving animals had recovered by day 4.
All females found dead showed body weight loss on the day of death. All surviving animals showed body weight gain over the study period.

Gross pathology:

Macroscopic post mortem examination of the animals that died during the study or surviving animals at termination did not reveal any histopathological changes that were considered to have arisen as an effect of treatment. Minor changes noted in each group were:
5000 mg/kg body weight: Black/brown stomach contents, dilatation of the renal pelvis
2800 mg/kg body weight: Dilatation of the renal pelvis
1580 mg/kg body weight: No abnormalities

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP regulation (EC) no. 1272/2008

Conclusions:

The oral LD50 values for male and female rats combined or for males separately were estimated as exceeding 5000 mg/kg. The estimated LD50 value for females alone was 4047 mg/kg.

Executive summary:

The purpose of this study was to assess the toxicity of the test article when administered to rats as a single oral dose.

The study was carried out in accordance with OECD Guideline No. 401, "Acute Oral Toxicity" and EEC Directive 84/449/EEC, Part B.1, "Acute Toxicity-Oral".

The substance was administered by oral gavage to five rats of each sex per group, at 5000 mg/kg body weight. As 4 females died during the study period 2 additional groups, each comprising of 5 females, were dosed at 2800 or 1580 mg/kg body weight. Macroscopic examination was performed at the end of the experimental period. Four females receiving 5000 mg/kg body weight died within 24 hours of dosing. No mortalities occurred during the study period among males receiving 5000 mg/kg or females receiving 2800 or 1580 mg/kg.

Signs of toxicity noted among animals receiving 5000 or 2800 mg/kg included piloerection, lethargy, hunched posture, ataxia and alopecia. Females receiving 1580 mg/kg showed no changes in clinical appearance. Females found dead on day 2 showed body weight loss. All surviving animals showed body weight gain over the study period. Macroscopic post mortem examination of animals found dead or surviving animals at termination did not reveal any abnormalities that were not commonly noted among rats of this age and strain or that were considered toxicologically significant.

The oral LD50 values for male and female rats combined or for males separately were estimated as exceeding 5000 mg/kg. The estimated LD50 value for females alone was 4047 mg/kg. Based on these results and according to the EEC criteria for classification and labelling requirements for dangerous substances (EEC Directive 83/467/EEC, Annex VI of the EEC Directive 67/548/EEC), the substance cannot be classified and has no obligatory labelling requirement.