Trisodium [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)][2-hydroxy-3-[(2-hydroxy-1-naphthyl)azo]-5-nitrobenzenesulphonato(3-)]chromate(3-)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)][2-hydroxy-3-[(2-hydroxy-1-naphthyl)azo]-5-nitrobenzenesulphonato(3-)]chromate(3-)

EC Number:

274-230-7

EC Name:

Trisodium [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)][2-hydroxy-3-[(2-hydroxy-1-naphthyl)azo]-5-nitrobenzenesulphonato(3-)]chromate(3-)

Cas Number:

69943-66-4

Molecular formula:

C32H18CrN8O14S2Na3

IUPAC Name:

Trisodium [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulphonato(3-)][2hydroxy-3-[(2-hydroxy-1-naphthyl)azo]-5-nitrobenzenesulphonato(3-)]chromate(3-)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Velaz Prague.
- Weight at study initiation: about 150 g.
- Housing: semparated, in plastic polypropylene cages T4 (supplied by s.p. Velaz Prague).
- Diet: animals were fed with commercial granular food mixture Altromin 1320, supplied by s.p. Velaz Prague. Daily dose of 15 g/animal/day.
- Fasting period before study: the day before of the test, animals were not fed.
- Water: CSN 757111 ad libitum.
- Bedding: wood shavings, from light wood.
- Acclimation period: 1 week.
Cleaning and disinfection of premises menagerie were made at dates determined, according to the standard operating procedures and compliance regime measures.

ENVIRONMENTAL CONDITIONS
- Temperature: controlled temperature at 22 ± 3 °C.
- Humidity: 50 ± 15 %.
- Photoperiod: 12 hrs dark / 12 hrs light, by fluorescent lamp.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20 %

Doses:

3981, 5020, 6310, 7943, 10000 mg/kg

No. of animals per sex per dose:

10 rats x dose

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: immediately after application, ca 30 minutes, after 3 hours post-application. The day after the application the observations were made in the morning and in the afternoon, while in the following days at least once daily. Body weight was measured at the beginning and end of the experiment.
- Necropsy of survivors performed: yes; internal organs were assessed for colour, size, consistency and structure.
- Other examinations performed: appearance of the skin, hair, visible mucous membranes status, mental activity, somatomotor activity, reactivity to stimulus, lacrimation, respiration, digestion, urogenital and circulatory apparatus. Organs and muscles were examined macroscopically. If post-mortem the bladder of animals was full, the urines were analyzed focusing on the detection of proteins, blood sugar, ketones, bilirubin, urobilinogen and pH.

Statistics:

The LD50 is calculated according to the probit method by Blisse.
Data on mortality, the frequency and using the logarithmic dose levels used, were inserted into a computer and analyzed by the PROBIT program, from October 1991 VUOS.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 6 620.9 mg/kg bw

Based on:

test mat.

Mortality:

3981 mg/kg: 0.0 %
5020 mg/kg: 30.0 %
6310 mg/kg: 30.0 %
7943 mg/kg: 70.0 %
10000 mg/kg: 100.0 %

Clinical signs:

other: After 3 hours post-application at dose level of 10.00 g/kg the following symptoms were recorded: - the appearance of skin and hair: stained with excretions. - nutritional status: cachectic. - appearance of visible mucous membranes: in the range of physiol

Other findings:

Autopsies of dead rats administered at dose of 1000 mg/kg revealed:
- External examination of the cadaver
Appearance of hair, skin: hair contaminated with excretions.
Nutritional status: cachectic.
Natural body openings: no macroscopical, patomorphological changes.
Integrity of the body surface: intect.
Postmortality: rigor mortis.

- Organs of head and neck: no macroscopical, patomorphological changes.

- Internal inspection cadaver
Tongue, esophagus, trachea: no macroscopical, patomorphological changes.
Lungs: pink, spongy consistency, airy, without macroscopical, patomorphological changes.
Heart: brownish-red colour, stiffer consistency, without macroscopical, patomorphological changes.

- Abdominal organs
Stomach: acute dilatation, filled with mash content, wall colour brown.
Guts: filled with sparse content, brown-coloured wall
Liver: brownish-red color, stiffer consistency, without macroscopical, patomorphological changes.
Spleen: stiffer consistency, red colour, without macroscopical, patomorphological changes.
Kidney: brownish-red colour, stiffer consistency, without macroscopical, patomorphological changes.
Peritoneum: no macroscopical, patomorphological changes.
Bladder: empty, without macroscopical, patomorphological changes.

- Urine - biochemical tests not established

Symptoms recorded at the lower dose level administered, are analogous to those of the maximum dose.

Any other information on results incl. tables

Overview of the time interval after the onset of various levels of logarithmic dose

Dose mg/kg	Percent mortality
	Day 1	Day 2	Day 3	Day 4	Day 5	Day 6	Day 7	Day 8	Day 9	Day 10	Day 11	Day 12	Day 13	Day 14
	1/2 hr	3 hrs
3981	0	0	0	0	0	0	0	0	0	0	0	0	0	0
5020	0	30	0	0	0	0	0	0	0	0	0	0	0	0
6310	0	20	10	0	0	0	0	0	0	0	0	0	0	0
7943	0	70	0	0	0	0	0	0	0	0	0	0	0	0
10000	0	70	0	0	0	0	0	0	0	0	0	0	0	0

Overview of mortality by sex of experimental rats at different levels

Mortality for each experimental group of tests for acute oral toxicity.

Dose mg/kg	Number of dead animals		Percentage
	Males	Females
3981	0	0	0
5020	1	2	30
6310	0	3	30
7943	4	3	70
10000	5	5	100

Body weight

Dose mg/kg	Males g/kg			Females g/kg
	Start test	End test	Difference	Start test	End test	Difference
3981	166.4	173	+ 6.6	160	168	+ 8
5020	167.6	178.8	+ 11.22	154.4	158.8	+ 44.4
6310	160.8	171.6	+ 10.8	159.6	154.8	- 4.8
7943	156	148.8	- 7.2	163.2	166	- 3.2
1000	162.8	163.2	- 9.6	148.8	148.4	- 0.4

Results

Dose g/kg	Group	Percentage of mortality	Log. Dose	Probits Exp.
3981	10	0.0	0.59999
5020	10	30.0	0.70070	4.476
6310	10	30.0	0.80003	4.476
7943	10	70.0	0.89998	5.524
10000	10	100.0	1.00000

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

Oral LD50: 6620.9 mg/kg (6098.6 - 7187.9 mg/kg).

Executive summary:

The acute toxicological characterization of the substance was determined by Acute toxicity test, according to the OECD guideline 401.

Groups of ten rats (5 males and 5 females) were administrated by oral gavage with 3981, 5020, 6310, 7943, 10000 mg/kg of test material. The LD50 was calculated according to the probit method by Blisse.

Conclusion

The LD50 was calculated to be 6.6209 g/kg (6.0986 - 7.1879 g/kg).