{[(2,4 and 2,5 and 2,6)-dimethylphenyl and 2-ethylphenyl]amino}{[(2,4 and 2,5 and 2,6)-dimethylphenyl and 2-ethylphenyl]amino}methaniminium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate

Endpoint summary

Description of key information

Acute oral toxicity, OECD Guideline 423, GLP compliant.

Acute inhalation toxicity, no study available

Acute dermal toxicity, study scientifically not necessary / other information available

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 29 July 2014 to 19 August 2014

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.

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

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 173 g and 202 g
- Fasting period before study: Food removed on D-1 and redistributed 4 hours after the test administration
- Housing: Solid-bottomed clear polycarbonate cages with a stainless steel mesh lid, containing sawdust bedding changed at least 2 times a week. Cages are placed in conventional air contitioned animal husbandry.
- Diet : Food stuff (A04, SAFE) supplied ad libitum
- Water : Tap water from public distribution system (microbiologically and chemically analysed every 6 months) supplied 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): at least 10 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2.0054 g of test item for the first step and 0.3068 or 0.3005 g of test item for the second and third step in 10 mL volumetric flask field of olive oil.
- Amount of vehicle (if gavage): 10 mL/kg/bw of preparation

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 (prior administration), D2, D7 and D14.
- Necropsy of survivors performed: yes. Animals anaesthetised with sodium pentobarbital and administration continued to fatal levels. Examination of organs likely to be modified in cases of acute toxicity.
- Other examinations performed: Observation and mortality report carried out every day : examination of behavioural or toxic effects on the major physiological functions: clinical signs (spontaneous activity, Preyer's reflex, respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, pupil appearance, salivation, lachrymation, righting reflex, back hair appearance)

Statistics:

no statistic was used

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Step 1 at 2000 mg/kg/bw: death of 2 of the rats at 22 hrs 55 minutes post-dose (2/3).
Step 2 and 3 at 300 mg/kg/bw : no mortality occured (6/6).

Clinical signs:

At 2000 mg/kg/bw of the 2 dead rats: rigor mortis before necropsy (2/2). Orange faeces and urine during D1 (2/2)

At 2000 mg/kg/bw of the surviving animal: no clinical signs (1/1). Orange faeces and urine during D1 (1/1).

At 300 mg/kg/bw : no clinical signs (6/6).

Body weight:

At 2000 mg/kg/bw of the surviving animal : evolution remained normal during the study (1/1)

At 300 mg/kg/bw : evolution remained normal during the study (6/6).

Gross pathology:

At 2000 mg/kg/bw: No treatment related changes revealed at the macroscopic examination (3/3).

At 300 mg/kg/bw: No treatment related changes revealed at the macroscopic examination (6/6).

Control study N° TA0 -2014 -006 on Olive Oil 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.

Performed from 08 July 2014 to 22 July 2014.

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 control item Olive Oil, 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)

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information 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 1000 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-F 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 2 rats treated at 2000 mg/kg/bw at 2 hrs 55 minutes post-dose (2/3).

Orange faeces and urine was noted during the 1st day of the test.

Rigor mortis was noted before the necropsy (2/2).

The macroscopic examination did not reveal treatment related changes (2/2).

In the surviving animal treated at 2000 mg/kg b.w. (1/3), orange faeces and urine was noted during the 1st day of the test.

The body weight evolution remained normal during the study.

The macroscopic examination of this animal at the end of the study did not reveal treatment related changes.

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

No clinical signs related to the administration of the test item at 300mg/kg were observed during the study.

The body weight evolution remained normal during the study.

The macroscopic examination of the animals at the end of the study did not reveal teatment 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 1000 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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

Moderate quality of the all database

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
According to OECD Series on Testing and Assessment No 237, reviews comparing the classification of oral and dermal hazards indicate that it is rare for the dermal test to yield a more severe classification. under this premise, substance would be classified in the corresponding GHS category as the oral hazard, if the LD50 oral is less than 300 mg/kg bw.

However, for substances where the oral LD50 range is between 300 and 2000 mg/kg and dermal penetration data indicates low dermal absorption (<10%) relative to oral absorption, the LD50 would equate to a dermal-equivalent value of 3000 mg/kg bw. However, this approach, which is assuming a high oral bioavailability, must be reconsidered if available information indicates low oral bioavailability of the substance.
Although the LD50 of the substance by oral route is between 300 and 2000 mg/kg, the toxicokinetic of the substance is not compliant with the above assumptions. Therefore, as a reasonable worst case scenario, and taking into account animal welfare consideration, the substance will be classified for dermal toxicity in the corresponding GHS category as the oral hazard, i.e. category 4, in the same way as the oral hazard was less than 300 mg/kg.
This is supported by the fact that in a sensitization test performed on the very closed analogue, no systemic toxicity was observed throughout the study on mice.

Endpoint conclusion

Endpoint conclusion:

no study available

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

Moderate quality of the whole database

Additional information

No dermal acute toxicity test is available. According to OECD series on Testing & Assessment No. 237, a waiving has been drawned based on the oral toxicity test already available on the substance. As a worst case scenario, the substance is classified for dermal toxicity in the corresponding GHS category as the oral hazard, i.e. category 4, in the same way as the oral hazard was less than 300 mg/kg, corresponding to a LC50 between 1000 and 2000 mg/kg. For the CAS, a LD 50 of 1001 mg/kg bw was therefore used.

Justification for classification or non-classification

Based on the available data, the substance is classified acute toxicity oral and dermal category 4.