[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2011-12-06 to 2011-12-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

EC Number:

265-449-9

EC Name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Cas Number:

65113-55-5

Molecular formula:

C25H30N3.C18H14N3O3S

IUPAC Name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: elevage JANVIER (53940 Le Genest St Isle - FRANCE)
- Age at study initiation:8 weeks old
- Weight at study initiation: between 181 g and 199 g
- Fasting period before study: Drinking water (tap water from public distribution system) and foodstuff (M20, SDS) were supplied freely. Food was removed at D-1 and then redistributed 4 hours after the test item administration
- Housing: Animal were housed by group of 3 in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Cage was installated in conventional air conditioned animal husbandry
- Diet (e.g. ad libitum): Ad libitum.
- Water (e.g. ad libitum):Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12h dark/12h light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: adapted for each animals
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: Olive oil was chosen among various vehicules because it does not induce pain. It has been chosen since it allowed to prepare a homogenous mixture, under stirring, usable for oral administration


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Based on previous examination of similar molecules, the dose of 2000 mg/kg was chosen.

Doses:

The 2 treated groups were dosed at 2000 mg/kg body weight

No. of animals per sex per dose:

3 females/dose/step:
STEP 1: 3 females at 2000 mg/kg body weight (N° Rf 9009, 9010 and 9011)
STEP 2: 3 females at 2000 mg/kg body weight (N° Rf 9048, 9049 and 9050)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily observations (spontaneous activity, preyer's reflex, respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, pupil appearance, salivation, lachrymation, righting reflex, back hair appearance) and weighing on days D0 (before administration), D2, D7 and D14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Macroscopic examinations at the end of the study (after anaesthesia) with the examination of organs likely to be modified in cases of acute toxicity (i.e. oesophagus, stomach, the entire digestif tract, spleen, liver, thymus, trachea, lungs, heart, kidneys, urinary bladder, ovaries, uterus, treatment area, adrenals, pancreas).

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured during the study

Clinical signs:

other: At T0 + 1 hour: STEP 1: blue faeces in all the treated animals / STEP 2: Rf 9049 and 9050 = blue faeces. Rf 9048 = decrease in spontaneous activity. At T0 + 3 and 4 hours: Blue faeces in all the treated animals in both step. D1 to D10: STEP 1: blue fae

Gross pathology:

The macroscopical examination of the animals at the end of the study revealed a white thickening of the forestomach in all the treated animals of the STEP 2.
No other observations or abnormalities were to report.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat.
In accordance with the OECD guideline N° 423, the LD50 cut-off of the test item may be considered to be higher than 2000 mg/kg body weight by oral route in the rat.
According to the classification, packaging and labelling of dangerous substances and preparations in accordance with the EEC directives 67/548, 2011/59 and 99/45, the test item must not be classified. No symbol or risk phrase is required.

In accordance with the regulation EC N° 1272/2008 on classification, labelling and packaging of substances and mixtures, the test item must not be classified. No signal word or hazard statement is required.

Executive summary:

The test item was administered to a group of 6 female Sprague Dawley rats at the single dose of 2000 mg/kg body weight. The experimental protocol was established according to the official method as defined in the OECD guideline N° 423 dated December 17th, 2001 and the test method B.1tris of the Council regulation N°440/2008.

 

No mortality occured during the study.

 

From the first day of the test, decrease in spontaneous activity (3/6) and blue faeces (6/6) were observed. The animals recovered a normal spontaneous activity on day 1 (2/3) and on day 10 (1/3), and recovered normal faeces on day 11 (3/6) and on day 13 (3/6).

 

A decrease in body weight of two animals (2/6) was noted on day 7: between -9% and -19% compared to day 0.

 

The macroscopical examination of the animals at the end of the study revealed a white thickening of the forestomach (3/6).

 

In conclusion, the LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat.

In accordance with the OECD guideline N° 423, the LD50 cut-off of the test item may be considered to be higher than 2000 mg/kg body weight by oral route in the rat.

According to the classification, packaging and labelling of dangerous substances and preparations in accordance with the EEC directives 67/548, 2011/59 and 99/45, the test item Solvent Black 46 must not be classified. No symbol or risk phrase is required.

 

In accordance with the regulation EC N° 1272/2008 on classification, labelling and packaging of substances and mixtures, the test item must not be classified. No signal word or hazard statement is required.