Chromate(3-), [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)][3-hydroxy-4-[(2-hydroxy-1-naphthalenyl)azo]-7-nitro-1-naphthalenesulfonato(3-)]-, sodium

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 06 to March 19, 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Chromate(3-), [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)][3-hydroxy-4-[(2-hydroxy-1-naphthalenyl)azo]-7-nitro-1-naphthalenesulfonato(3-)]-, sodium

EC Number:

284-915-2

EC Name:

Chromate(3-), [3-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-2-hydroxy-5-nitrobenzenesulfonato(3-)][3-hydroxy-4-[(2-hydroxy-1-naphthalenyl)azo]-7-nitro-1-naphthalenesulfonato(3-)]-, sodium

Cas Number:

84989-26-4

Molecular formula:

C36H20CrN8Na3O14S2

IUPAC Name:

trisodium 12'-methyl-6',7-dinitro-14'-phenyl-2',13,15,16'-tetraoxa-1λ⁵,2,9',10'λ⁵,13',14'-hexaaza-14-chromaspiro[hexacyclo[12.11.0.0³,¹².0⁴,⁹.0¹⁶,²⁵.0¹⁹,²⁴]pentacosane-14,1'-tetracyclo[8.6.0.0³,⁸.0¹¹,¹⁵]hexadecane]-1,3,3'(8'),4',5,6',7,9,9',11,11'(15'),12',16(25),17,19,21,23-heptadecaene-1,10'-bis(ylium)-14,14,14-triuide-4',10-disulfonate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintier farm Madoerin AG, CH-4414 Fuellinsdorf/Switzerland.
- Age at study initiation: 9 to 11 weeks.
- Weight at study initiation: males 201 - 245 g and female 178 - 212 g.
- Fasting period before study: fasted for 12 to 18 hours (access to water was not interrupted). Food was again presented approximately one hour after dosing.
- Housing: groups of five in Makrolon type-3 cages with standard softwood bedding. The cages were cleaned twice weekly during the test period.
- Diet: ad libitum; pelleted standard Kliba 343, Batch 36/85 and 39/86 rat maintenance diet. Food was analysed for contaminants.
- Water: ad libitum; community tap water from Itingen. Water was analysed for contaminants.
- Acclimation period: at least one week under laboratory conditions, after veterinary examination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C.
- Humidity: 40 - 70 %.
- Air changes: air-conditioned with 10-15 air changes per hour.
- Photoperiod: 12 hours artificial fluorescent light/12 hours dark, music/light period.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

TEST ARTICLE PREPARATION
The test article was placed into a glass beaker on a tared Mettler PK 4800 balance, and the vehicle was added.
A weight by volume dilution was prepared using a homogenizer (Ultra-Turrax, Janke and Kunkel, Staüfen, FRG).
Homogeneity of the test article in the vehicle was maintained during treatment using a magnetic stirrer.
The preparation was made immediately prior to dosing.

VEHICLE
4 % solution of CMC, carboxymethylcellulose sodium salt purum (Fluka AG, Buchs / Switzerland in distilled water).

Doses:

Application Volume/kg body weight:
10 ml at 1000 mg/kg
20 ml at 3000 mg/kg
20 ml at 5000 mg/kg
20 ml at 8000 mg/kg

No. of animals per sex per dose:

5 males and 5 females per group

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: mortality/viability were recorded four times during test day 1, and daily during days 2-15. Body weights were recorded at the test days 1 (pre-administration), at the day 8 and the day 15.
- Necropsy of survivors performed: yes. Necropsies were performed by experienced prosectors. All animals were necropsied. All animals surviving to the end of the observation period were killed by intraperitoneal injection of sodium pentobarbitone.
- Other examinations performed: each animal was examined for changes in appearance and behaviour four times during day 1, and daily during days 2-15.All abnormalities were recorded.

Statistics:

The LOGIT-Model (COX, Analysis of Binary Data, London 1977) was applied to estimate the toxicity value. Additionally, the 90, 95 and 99 % confidence intervals for the toxicity and the slope of the concentration response line were estimated.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 792 mg/kg bw

Based on:

test mat.

95% CL:

3 813 - 14 980

Mortality:

1000 mg/kg: no deaths occurred.
3000 mg/kg: one female dead at day 2 and two females dead at the day 3; no males dead.
5000 mg/kg: two females dead at day 2 and one at the day 3; no males dead.
8000 mg/kg: three females dead at day 2 and one at the day 3; two males dead at day 2 and one at the day 3.

Clinical signs:

other: The following symptoms were observed: 1000 mg/kg: sedation, dyspnea, curved body position, ruffled fur. 3000 mg/kg: sedation, dyspnea, ataxia, curved body position, diarrhea, ruffled fur. 5000 mg/kg: sedation, dyspnea, ataxia, curved body position, diarrh

Gross pathology:

1000 mg/kg killed:
lung: partly red, focal (2), partly red, maculate (1), dark-red, mottled (3), no pathologic changes (4)
3000 mg/kg dead:
lung: black, mottled (1), reddish foci/4 mm (1)
liver: black discoloured (1), black (2)
stomach/intestines : black (3)
anal region: black (1)
3000 mg/kg killed:
lung: mottled (3)
no pathologic changes (4)
5000 mg/kg dead
lung: mottled (1)
stomach/intestines : black (2), black contents (1)
5000 mg/kg killed
no pathologic changes (7)
8000 mg/kg dead
liver: black discoloured (7)
lung : mottled (5)
stomach/intestines: black (7)
8000 mg/kg killed
no pathologic changes (3)

Applicant's summary and conclusion

Interpretation of results:

other: Not classified as harmful or toxic according to the CLP Regulation (EC) No. 1272/2008.

Conclusions:

LD50: 5792 mg/kg bw (CL 95 %: 3813 - 14980 mg/kg).

Executive summary:

Method

The test article was administered to rats of both sexes by oral gavage, at doses from 1000 to 8000 mg/kg. The study procedures followed fulfil the OECD guideline 401 and EU method B1.

Results

The following death rate was observed:

0 % at 1000 mg/kg;

30 %. at 3000 mg/kg;

30 % at 5000 mg/kg;

70 % at 8000 mg/kg.

Based on these observations, the Logit-Estimation for the acute oral toxicity in rats of both sexes observed for a period of 15 days is 5792 mg/kg with 95 % CI of males/females 3813 - 14980 mg/kg