Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

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
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): Sepisol fast Blue 85219
- Substance type: organic salt - UVCB
- Appearance/Physical state: dark blue powder
- Stability under test conditions: stable
- Storage condition of test material:room temperature
Specific details on test material used for the study:
- Batch N° 310551

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:

TEST ANIMALS
- Source: élevage Janvier, Le Genest-St-Isle, France
- Age at study initiation: approximately 7 weeks
- Weight at study initiation:
At d1, mean for the 2000 mg/kg group: 221.1 gram +/- 8.8
At d1, mean for the 300 mg/kg group: 223.3 gram +/-8.6
- Fasting period before study: deprived of water overnight prior to dosing
- Housing: polypropylen cage. Maximum 3 animals per cage
- Diet : ad libitum except the 4 hours following administration. granules A04-10
- Water: ad libitum
- Acclimation period: 5 days at least before administration

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 50 +/- 20
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 hrs/12hrs

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: adapted for each animal
- Amount of vehicle : 5 mL/kg for both doses
- Justification for choice of vehicle: corn oil was chosen among various vehicules because it does not induce pain. Following several preliminary
assays, the aqueous-based vehicles (pure water, HCMC) were disregarded and the non-polar vehicle (corn oil commonly used for oral toxicity assay ) has been chosen since it allowed to prepare a homogenous mixture usable for oral adiministration.
- Lot/batch no. : 07030169/C

Doses:
2000 mg/kg
300 mg/kg
No. of animals per sex per dose:
6 rats for the 2000 mg/kg dose
6 rats for the 300 mg/kg dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: regularly following administration (30 min, 1h, 2h, 3h and 4 h) and at least once a day during the 14 days
- Frequency of weighing: d-1, d1 T0, d4, d8, d15
- Necropsy of survivors performed: yes

Results and discussion

Preliminary study:
first dose of 2000 mg/kg with 3 animals
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
At the 300mg/kg dose tested, no deaths were observed.
At the 2000 mg/Kg dose: deaths occurred for all animals between day 2 and day 6.
Clinical signs:
At the 2000 mg/kg dose, significant symptoms occured: important piloerection, reduced motor activity with diarrhoea and hypotonia among one female.
At the 300 mg/kg dose, the test materai has caused piloerection associated with a reduced motor activity during the first 4 hours but no deaths occured.
Body weight:
At the 300 mg/kg: the mean body weight observation has not shown any sign associated with the test material.
At the 2000 mg/kg: lost of weight has been observed for two animals before their death
Gross pathology:
At the 2000 mg/kg dose: the test material (colorant) has been found to be present all along the digestive tract and also in the main organs
(kidneys, liver, spleen, blader, pancreas) for the 6 females.
At the 300 mg/kg dose: no tissue neither organic abnormalities have been found for all 6 females.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
According to the Global Harmonised System (GHS), the test material Sepisol Fast Blue 85219 has been classified as acute toxicity category 4 with a LD 50 determined between 2000 and 300 mg/kg in rat.
Executive summary:

The aim of this study was to assess qualitatively and quantitatively the toxic effects and the delay of the appearance after single oral administration of pre-defined doses of 2000 mg/kg and 300 mg/kg of body weight, of test element named SEPISOL FAST BLUE 85219 suspended in corn oil, in 6 females rats for each dose level, using a stepwise procedure. The study has been performed by using the OECD guideline 423.

The animals were daily observed for at least 14 days after administration and the clinical signs and signs of toxicity were noted.

A preliminary test was performed with a 2000 mg/kg dose after which all animals died.

The results of the assay showed that 6 animals died with the 2000 mg/kg dose level (100 % of mortality) and that no animals died with the 300 mg/kg dose level (0% of mortality).

At the 2000 mg/kg dose, significant symptoms occured: important piloerection, reduced motor activity with diarrhoea and hypotonia among one female. At the 300 mg/kg dose, the test materai has caused piloerection associated with a reduced motor activity during the first 4 hours but no deaths occured. At the 2000 mg/kg dose: the test material (colorant) has been found to be present all along the digestive tract and also in the main organs (kidneys, liver, spleen, blader, pancreas) for the 6 females. At the 300 mg/kg dose: no tissue neither organic abnormalities have been found for all 6 females.

This test provided results allowing the test element to be classified according to the Classification and Labeling of Products regulation (CLP EC 1272/2008) , which cause acute toxicity.

The test element was classified in the hazard category 4 with a LD50 ranging between 2000 mg/kg and 300 mg/kg