Disodium 4,4'-bis[[4-anilino-6-[(2-hydroxyethyl)amino]-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Endpoint summary

Description of key information

Rat oral LD50 > 5000 mg/kg bw

Rat inhalation LC50 > 1895 mg/m3

Rat dermal LD50 > 2000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines. Justification for Read Across is detailed in the endpoint summary and in the Category Justification Report attached to the section 13.

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

limited data

GLP compliance:

not specified

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Stain: Albino rat, strain not specified.
- Age at study initiation: young

Route of administration:

oral: gavage

Vehicle:

not specified

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

3 x sex x dose

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, histopathology

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality observed

Clinical signs:

other: Dyspnoea, exophthalmoses, ruffled fur, and curved body position were seen, being common symptoms in acute toxicity testing. The animals recovered within 11 days.

Gross pathology:

At autopsy, no deviations from normal morphology were found.

Interpretation of results:

not classified

Remarks:

Migrated information According to the CLP regulation. Criteria used for interpretation of results: EU

Conclusions:

LD50 > 5000 mg/kg bw.

Executive summary:

Method

The exploratory (approximate) acute oral LD50 for test substance was investigated by administering a single dose of 5000 mg/kg bw by gavage to the young adult albino rats.

Results

LD50 > 5000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Particle size not specified. Justification for Read Across is detailed in the endpoint summary and in the Category Justification Report attached to the section 13.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann, Borchen, Germany.
- Weight at study initiation: 180 - 200 g.

Route of administration:

inhalation: dust

Type of inhalation exposure:

not specified

Vehicle:

air

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetric with membrane filter.

Duration of exposure:

1 - 4 h

Concentrations:

163.3, 375, 1225 and 1895 mg/m³ air at 4 hour exposure.
1820 mg/m³ air at 1 hour exposure.

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days .
- Necropsy of survivors performed: yes.
- Other examinations performed: clinical signs.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1 895 mg/m³ air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Sex:

male/female

Dose descriptor:

LC50

Effect level:

1 820 mg/m³ air (nominal)

Based on:

test mat.

Exp. duration:

1 h

Mortality:

No mortality occuerred.

Clinical signs:

other: At concentrations of 1225 and 1895 mg/m³ air at the 4 hour exposure the animals showed a decreased general condition for about 4 - 6 hours.

Gross pathology:

No abnormalities detected.

Interpretation of results:

other: not applicable.

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

LC50 (4 h): > 1.895 mg/l air

Executive summary:

Method

Acute inhalation toxicity was assessed following the method described into the OECD guideline 403.

Results

LC50 (4 h): > 1.895 mg/l air

LC50 (1 h): 1.820 mg/l air

Conclusion

According to CLP Regulation (EC 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The inhalation LD50 value classification limit for dust is 5.0 mg/l (category 4: 1.0 < ATE ≤ 5.0 mg/l). In the current test an LD50 was non identified; considering the fact that no mortality occurred, a classification category can not be assigned. Thus, a classification according to the CLP Regulation (EC 1272/2008) is not applicable.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

1 895 mg/m³ air

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Only a summary is available;. Justification for Read Across is detailed in the endpoint summary and in the Category Justification Report attached to the section 13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

no

Remarks:

Pre GLP.

Test type:

standard acute method

Species:

rabbit

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: mean 2205 g.
- Housing: one animal per cage.
- Food: Nafag Würfel Nr. 84.

ENVIRONMENTAL CONDITIONS
- Temperature: 23°C ± 2°C
- Humidity: 55 ± 5%
- Photoperiod: 14 hrs light/day.

Type of coverage:

occlusive

Vehicle:

other: fresh suspension with gum arabic 1% / tap water

Details on dermal exposure:

APPLICATION
One application to intact skin. Occlusive dressing (Draize method) for 24 hours.

TEST SITE
- Area of exposure: dehairing of the dorsal skin by shearing.
- % coverage: SURFACE OF 200-300 cm2

REMOVAL OF TEST SUBSTANCE
- Washing: washing the application area with lukewarm water and a sponge.

TEST MATERIAL
- Concentration: 80%

Duration of exposure:

24 hours

No. of animals per sex per dose:

3 males and 3 females

Details on study design:

- Duration of observation period following administration: 8 days.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred.

Clinical signs:

other: No systemic symptoms were recorded.

Skin reactions recorded

Dose mg/kg	Time point	N. of animals with reactions	Reaction
2000	24 hrs	6/6	Erythema
	2 days	1/6	Erythema, oedema
	3 days	1/6 1/6	Erythema Oedema
	4 days	1/6 1/6	Erythema Scaling, oedema
	5 + 6 days	1/6	Scaling, oedema
	7 + 8 days	2/6 1/6	Scaling Scaling, oedema

Interpretation of results:

not classified

Remarks:

Migrated information According to the CLP regulation. Criteria used for interpretation of results: EU

Conclusions:

LD50 > 2000 mg/kg bw

Executive summary:

Method

The acute dermal LD50 of the test item was tested in rabbits administed by dermal occlusive dressing (Draize method) at a single concentration of 2000 mg/kg bw.

Results

No deaths occurred.

LD50 > 2000 mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Many summaries of tests performed according to official guidelines are available on the analogous substance CAS 16324-27-9 of the Stilbene Fluorescent Whitening Agents category, assessing a LD50 > 10000 mg/Kg bw. The two substances share the same organic functional group and substitution on the triazino moyety with monohydroxyethylamino. They are therefore very similar both according to the chemical structure (Tanimoto similarities > 90 %) than according to metabolic pathway (OECD Toolbox profiling). The two substances differ in the sulphonation degree: the substance under registration (CAS 17958-73-5) is the disulphonate derivative, while CAS 16324-27-9 is tetrasulphonated. The difference in solubility is of almost one order of magnitude (30.2 g/l vs 140 g/l), but this difference has no impact from a toxicological point of view, because both substances are in any case highly soluble, stable and completely dissociated in water.

Further studies are reported the results for the analogous substance CAS 4193-55-9, which has a LD50 > 5000 mg/Kg bw.

This substance is the analogous disulphonated, dihydroxyethylamino substituted. In this case the suphonation degree is the same as that of CAS 17958-73-5, while the substitution on the triazino ring is different (dihydroxyethyl for CAS 4193-55-9, instead monohydroxyethyl for CAS 17958-73-5). From a metabolic point of view, the two substances can be considered very similar because the monohydroxyl derivative is a metabolite of the dihydroxy derivative. In this respect the water solubilities of CAS 17958-73-5 and CAS 4193-55-9 are more similar (30.2 vs 48.2 g/l, respectively) and the LD value of 5000 g/l assessed for CAS 4193-55-9 can be considered as conservative representative for the substance under registration.

A Klimish 2 study for acute dermal toxicity was performed at 2000 mg/Kg bw with no effect on CAS 16324 -27 -9, for which the same argumentation described for acute oral toxicity are applicable.

Because of the physical state and the trade forms, inhalation is not an appropriate route of exposure. Acute toxicity results for the other two exposure routes indicate no concern therefore no testing was performed.One test is available on an analogous substance, performed at the maximum allowed concentration of 1890 mg/m³: no effects were observed.

Within the whole category eleven over fourteen registered substances were tested for acute oral toxicity and seven for dermal: none of the existing tests arisen any concern at the highest tested doses.

Justification for selection of acute toxicity – oral endpoint

Study conducted according to internationally accepted testing guidelines.

Justification for selection of acute toxicity – dermal endpoint

Study conducted according to internationally accepted testing guidelines, in GLP.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.1 Acute toxicity section, substances can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route according to the numeric criteria. Acute toxicity values are expressed as (approximate) LD50 (oral, dermal) or LC50 (inhalation) values or as acute toxicity estimates (ATE).

The oral LD50 value was established to be greater than 5000 mg/kg body weight, therefore the test substance is out of any classification limit for acute oral toxicity (oral acute toxicity category 4: 300 < ATE ≤ 2000 mg/kg bw).

The dermal LD50 value was established to exceed 2000 mg/kg body weight, which exceeded the highest CLP classification limit (dermal acute toxicity category 4: 1000 < ATE ≤ 2000 mg/kg bw).

The inhalation LC50 value was established to be greater than 1895 mg/m³. For powder the limit for classification is ATE > 5 mg/l i. e. 5000 mg/m³.

Since no effect was observed at the tested concentration and this was the maximum reachable concentration in the test condition, it is assumed that the substance is not classified for inhalation acute toxicity.

In conclusion, the available experimental data are adequate for classification and labelling and the test substance is non classified for oral and dermal acute toxicity, according to the CLP Regulation (EC 1272/2008).