Registration Dossier

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:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals 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:
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 faeces in all the treated animals / STEP 2: blue faeces in all the treated animals and Rf9049 = decrease in spontaneous activity from D8 to D10.

D11 to D12: STEP 1: blue faeces in all the treated animals / STEP 2: nothing to report

D13 to D14: Nothing to report in all the treated animals in both step.
Body weight:
A decrease in body weight of two animals (Rf 9048 and 9049 - STEP 2) was noted on day 7: between -9% and -19% respectively, compared to day 0.
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 Sepisol Black 46 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 Sovent 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.
Executive summary:

The test item Solvent Black 46 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 Sepisol Black 46 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.