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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 06 November 2012 to 27 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability is rated 1 because the study followed the standard guideline of reference (OECD 471), which describes a procedure designed to evaluate this endpoint, the results were reviewed for reliability and assessed as valid, and the study was conducted under GLP condition.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
anilino(phenylimino)methanaminium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
EC Number:
941-151-0
Molecular formula:
C31H28N6SO3
IUPAC Name:
anilino(phenylimino)methanaminium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Sepisol Fast Yellow MG-DPG
- Physical state: Power
- Stability under test conditions: Stable
- Storage condition of test material: room temperature

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 or 9 weeks old
- Weight at study initiation: between 196 g and 239 g
- Fasting period before study: Food removed on D-1 and redistributed 4 hours after the test administration
- Housing: Solid-bottomed clear poycarbonate cages with a stainless steel mesh lid, containing sawdust bedding changed once a week. Cages are placed in conventional air contitioned animal hyusbandryµ.
- Diet : Food stuff (A04, SAFE) is supplied ad libitum
- Water : Tap water from public distribution system (microbiologicaly and chemicaly analysed every 6 months) is supplie ad libitum.
- Acclimation period: Five days at least.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70% RH
- Air changes (per hr): 10 to 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
DMSO
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2.0219 g of test item for the first step and 0.3025 or 0.3013 g of test item for the second and third step in 10 mL of DMSO.
- Amount of vehicle (if gavage): 10 mL/kg/bw of preparation
- Justification for choice of vehicle: the use of distilled water or olive oil gives a suspension. Because the use of the DMSO gives a homogeous solution the DMSO was chosen as the vehicule of the study.

Doses:
Step 1: 2000 mg/kg/bw
Steps 2 and 3: 300 mg/kg/bw
No. of animals per sex per dose:
3 females per dose per step.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily examination. Weighing on D0 (prio administration), D2, D7 and D14.
- Necropsy of survivors performed: yes
- Other examinations performed: Observation and mortality report carried out every day : examination of behavioural or toxic effects on the major physiological functions:
* Body weight
* Clinical signs (spontaneous activity, Preyer's reflex, respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, mydriasis, salivation, lachrymation, righting reflex, piloerection, mortality)
Statistics:
no statistic was used

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
<= 300 mg/kg bw
Based on:
test mat.
Mortality:
Step 1 at 2000 mg/kg/bw: death of the rats at 3 hrs post-dose.
Step 2 and 3 at 300 mg/kg/bw : no mortality occured.
Clinical signs:
At 2000 mg/kg/bw: absence or decrease in spontaneous activity (3/3), in Preyer's reflex (1/3), in muscle tone (3/3) and in righting reflex (3/3), polypnea (1/3), dyspnea (2/3), myosis (2/3) and salivation (1/3) before death

At 300 mg/kg/bw during the first hour: absence or decrease in spontaneous activity (6/6), in muscle tone (3/6) and in righting reflex (3/6) , increase in mucle tone (3/6), salivation (1/6), lacrymation (1/6), chromodacryorrhea (1/6), partial prosis (1/6) and piloeretion (1/6).
Recovery of a normal behaviour between 24 and 72 hrs post-dose.

Body weight:
At 2000 mg/kg/bw : no data because of the death of the animals 3 hrs post-dose.

At 300 mg/kg/bw : lower body weight in the animals on day 2 compared to D0: +2% versus +13% in the historical control group. The animals recovered a normal body weight evolution on day 7.
Gross pathology:
At 2000 mg/kg/bw: The macroscopic examination revealed a thinnning of the forestomach (3/3), associated with an orange coloration of the forestomach (3/3) and red spots and orange coloration of the corpus (3/3).

At 300 mg/kg/bw: at the end of the study, no treatment related changes were observed at the macroscopic examination.

Any other information on results incl. tables

Control study N° TA0 -2012 -009 on DMSO used as the vehicule in the study.

The study was performed to assess the comportment of the strain of rat used at Phycher laboratory in its environment and to give additional historical data.

The method was designed to meet the requirements of the following:

- OECD guideline for the testing of chemicals N°423 dated December 17th, 2001

- Method B.1tris of the council regulation N°440/2008

Three animals received the DMSO, administered by gavage under a volume of 10 mL/kg body weight using a suitable syringe graduated fitted with an oesophageal metal canula.

No clinical signs, body weight changes nor treatment related changes were reported (3/3 animals normal)

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information according to the Regulation EC N° 1272/2008 Criteria used for interpretation of results: EU
Conclusions:
The LD50 of the test item is higher than 300 mg/kg/bw and lower than 2000 mg/kg/bw by oral route in rats.
In accordance with the OECD guideline N°423, the LD50 cut-off of the test item may be considered as 500 mg/kg/bw by oral route in the rat.

In accordance with the E.E.C directives 67/548, 2001/59 and 99/45, the test item must be classified R22 "harmful is swallowed". The symbol "Xn" and the warning label "Harmful" are required.
In accordant with the regulation EC 1272/2008, the test item must be classified in category 4. The signal word "Warning" and hazard statement H302 "Harmful is swallowed" are required.
Executive summary:

The test item Sepisol Fast Yellow MG-DPG was administered to a group of 3 female Sprague Dawley rats at the single dose of 2000 mg/kg/bw and then to a group of 6 females Sprague Dawley at the single dose of 300 mg/kg/bw. The experimental protocol was established according to the official method as defined in the OECD guideline No. 423 date December 17th, 2001, and the test method B.1tris of the Council regulation No. 440/2008.

It was noted the death of the 3 rats treated at 2000 mg/kg/bw at 3 hrs post-dose. The mortalities were preceded by absence or decrease in spontaneous activity (3/3), in Preyer's reflex (1/3), in muscle tone (3/3) and in righting reflex (3/3), polypnea (1/3) dyspnea (2/3), myosis (2/3) and salivation (3/3).

The macroscopic examination of the dead animals revealed a thinning of the forestomach (3/3) associated with an orange coloration of the forstomach (3/3) and red spots and orange coloration of the corpus (3/3).

No mortality occured in animals treated at 300 mg/kg/bw.

An absence or decrease in spontaneous activity (6/6), in muscle tone (3/6), and in righting reflex (3/6), increase in muscle tone (3/6), sakuvatuib (1/6), lacrymation (1/6), chromodacryorrhea (1/6), partial ptosis (1/6) and piloerection (1/6) were noted during the first hours of the test. the naimals recovered anormal behavior between 24 and 72 hrs post-dose.

A lower body weight gain was noted in the animals on day 2 comprated to day 0 :+2% versus +13% in the historical control group. The animals recovered a normal body weight evolution on day 7.

The macroscopic examination of the animals at the end of the study did not revel treatment related changes.

In conclusion, the LD50 of the test item is higher the 300 mg/kg/bw and lower than 2000 mg/kg/bw by oral route in rats.

In accordance with the OECD guidelin No. 423, the LD50 cut-off ot the test item may be considered as 500 mg/kg/bw by oral route in rats.

According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with the E.E.C directives 67/548, 2001/59 and 99/45, the test item must be classified R22 "Harmful is swallowed". The item must be characterised by the symbol "Xn" and the warning label "Harmful".

In accordance with the regulation EC No. 1272/2008, the test item must be classified in category 4. The signal word "Warning" and hazard statement H302 "Harmful is swallowed" are required.