tetrakis[({bis[4-(dimethylamino)phenyl] and 4-(methylamino)phenyl} or {4-(dimethylamino)phenyl and bis[4-(methylamino)phenyl]} or {tris[4-(dimethylamino)phenyl]}methylium)]C12-(branched and/or linear)-alkyl-(4-sulfonatophenoxy)benzenesulfonate and oxybis[C12-(branched and/or linear)-alkyl-benzenesulfonate].

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2008/03/18 to 2008/04/03

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 423), 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:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

bis(bis[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylium) bis(tris[4-(dimethylamino)phenyl]methylium) 3-(dodecan-3-yl)-4-(2-dodecyl-4-sulfonatophenoxy)benzene-1-sulfonate 3-dodecyl-4-(4-sulfonatophenoxy)benzene-1-sulfonate

EC Number:

700-759-4

Molecular formula:

not applicable

IUPAC Name:

bis(bis[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylium) bis(tris[4-(dimethylamino)phenyl]methylium) 3-(dodecan-3-yl)-4-(2-dodecyl-4-sulfonatophenoxy)benzene-1-sulfonate 3-dodecyl-4-(4-sulfonatophenoxy)benzene-1-sulfonate

Details on test material:

- Name of test material : Sepisol Fast Violet 881239
- Substance type: Organic salt - UVCB
- Physical state/ appearance: violet powder
- Lot/batch No.: 601204
- Storage condition of test material: ambient temperature, in the dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: élevage JANVIER (53940 Le Genest-St-Isle, France).
- Age at study initiation: approximately 8 weeks.
- Weight at study initiation: At D1, mean (for group 1 and 2): 223.9 gram +/- 6.3
- Fasting period before study: deprived of food overnight prior administration.
- Housing:Housed in polypropylene cage (31 cm x 46 cm x 19 cm) with a stainless steel lid. 3 animals per cage. Sterilized and dust free sawdust litter.
- Diet : ad libitumexcept the night prior administration and 3-4 hours following administration (sterilized granules A04-10).
- Water: ad libitum. Distributed by a polypropylene biberon with a stainless steel teat.
- Acclimation period:at least 5 days before dosing.

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

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2000 mg/kg of test item in corn oil. Administration volume of 5 ml/kg.
- 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. : Cooper - lot 07030169/C

Doses:

2000 mg/kg for both groups.

No. of animals per sex per dose:

3 rats for both groups (6 in total)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: regularly following administration (immediately, 30 min, 1h, 2h, 3h and 4 h) and at least once a day until the end of the study
- Frequency of weighing: D-1, D1/T0 just before administration, D4, D8, D15
- Necropsy of survivors performed: yes
- Other examinations performed:
* 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, grip strength, diarrhea, lethargy, coma, skin, fur and eyes changes, mortality)
* Autopsy

Results and discussion

Preliminary study:

First dose of 2000 mg/kg with 3 animals.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At the 2000 mg/kg dose: no deaths were observed for both groups.

Clinical signs:

other: At the 2000 mg/kg dose, symptoms occured: important piloerection and reduced motor activity which persisted maximum 24 hrs after administration of the test item. Colored feces on D1 for all 6 animals until D3 for 3/6 animals. No organic lesion or tissue

Gross pathology:

Nothing to report

Any other information on results incl. tables

Dose = 2000 mg/kg

Clinical observations

Time of observation	Comments
D1 (following administration)	Important piloerection for all animals.
D1 T30'	Important piloerection and reduced motor activity for all animals. Violet feces for 3/6 animals
D1 T1h	Important piloerection, reduced motor activity and violet feces for all animals
D1 T2h	Important piloerection, reduced motor activity and violet feces for all animals
D1 T3h	Important piloerection, reduced motor activity and violet feces for all animals
D1 T4h	Important piloerection, reduced motor activity and violet feces for all animals
D2	Animals 5644 to 5646: important piloerection, reduced motor activity and violet feces. Animals 5647 to 5649: Blue feces.
D3	Animals 5644 to 5646: blue feces. Animals 5647 to 5649: Nothing to report.
D4	Nothing to report
D5 to D15	Nothing to report

Autopsy

Animals N°		Mortalilty		Comments
		Day	Cause
1^stStep	5644	D15	Euthanasia	Nothing to report
	5645	D15	Euthanasia	Nothing to report
	5646	D15	Euthanasia	Nothing to report
2^dStep	5647	D15	Euthanasia	Nothing to report
	5648	D15	Euthanasia	Nothing to report
	5649	D15	Euthanasia	Nothing to report

Body Weight evolution

Animals N°		Weight (g)
		D1	D4	D8	D15	D15-D1
1^stStep	5644	219.4	245.6	258.4	278.7	59.3
	5645	217.2	246.7	274.1	304.3	87.1
	5646	230.5	242.9	274.7	290.4	59.9
2^dStep	5647	218.1	247.0	279.3	323.8	105.7
	5648	228.2	253.3	261.3	278.8	50.6
	5649	230.1	249.2	265.3	283.1	53.0
Mean (standard deviation)		223.9 6.3	247.5 3.5	268.9 8.4	293.2 17.8	69.3 22.1

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the CLP regulation, the test material Sepisol Fast Violet 881239 has not been classified as acute toxicity with a LD 50 determined to be above the threshold of 2000 mg/kg bw.

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 a pre-defined dose of 2000 mg/kg body weight, of a test element named SEPISOL FAST VIOLET 881239 suspended in corn oil, in 6 females rats, using a stepwise procedure. The experimental protocol was established according to the official method as defined in the OECD guideline No. 423 dated December 17th, 2001.

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

The first step of the assay was performed with a 2000 mg/kg dose after which no animals dies. So, the second step of the assay was also performed with a 2000 mg/kg dose, after which no animals dies.

Important piloerection, reduced motor activity which persisted 24 hrs after administration were observed

No organic lesion or tissue injury has been observed at the autopsy.

In conclusion, the LD50 of the test item Sepisol Fast Violet 881239 is higher than 2000 mg/kg body weight by oral route in rats.

In accordance with the OECD guideline No. 423, the LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in rats.

According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with the EEC directive 67/548, 2001/59 and 99/45, the test item Sepisol Fast Violet 881239 must not be classified. No symbol or risk phrase is required.

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