Tetrasodium 4,4'-bis[[4-[bis(2-hydroxyethyl)amino]-6-(4-sulphonatoanilino)-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:

experimental study

Adequacy of study:

key study

Study period:

From September 21 to October 5, 1981

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Tif:RAif (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7 to 8 weeks.
- Body weights at study initiation: males - 175 g; females - 176 g.
- Fasting period before study: overnight.
- Housing: in groups of 5 in Macrolon cages (type 3).
- Diet: rat food NAFAG No. 890 NAFAG Gossau SG, ad libitum.
- Water: tap water, ad libitum.
- Acclimation period: 4 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C.
- Humidity: 55 ± 10 %.
- Photoperiod: 12 hours light per day.

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

400

Details on oral exposure:

VEHICLE
- Amount of vehicle: 20 ml/kg body weight.

Doses:

5000 mg/kg body weight.

No. of animals per sex per dose:

5 animals/sex

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: physical condition and rate of deaths were monitored throughout the whole observation period; body-weights were recorded immediately prior to dosing (control weights) and at 7 and 14 days.
- Necropsy of survivors performed: yes; all animals were submitted to a necropsy, whenever they died, survivors at the end of the observation period.
- Other examinations performed: gross examination.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occured.

Clinical signs:

other: Dyspnoea, exophthalmos, ruffled fur reported, diarrhoea and curved body position were observed. Nevertheless all symptoms are recovered within 9 days.

Gross pathology:

No compound related gross organs changes were reported.

Signs and symptoms

hrs

Days

1

2

3

5

24

2

3

4

5

6

7

8

9

10

11

12

13

14

Sedation

Dyspnoea

+

+

+

+

+

+

+

+

+

+

+

+

Dacryorrhoea

Chromodacryorrhoea

Rinorrhoea

Epistaxis

Exophthalmos

+

+

+

+

+

+

+

Salivation

Ruffled fur

++

++

++

++

+

+

+

+

+

+

+

Pallor

Cyanosis

Diarrhoea

+

+

Body position (ventral)

Body position (lateral)

Body position (curved)

+

+

+

+

+

+

+

+

+

+

Ataxia

Trismus

Tremor

Tonic clonic muscle spasms

Convulsions

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 acute oral toxicity of the test compound in rats of both sexes was tested at the single concentration of 5000 mg/kg bw and observed over a period of 14 days.

Results

The LD50 is greater than 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:

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

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Particle size not specified. The analogous substance, part of the Stilbene Fluorescent Withening Agents, group 3, is the acid form of the disulphonated derivative dihydroxyethyl derivative, therefore contains the same organic functional groups, but due to the sulphonation/salification degree it is less soluble. This property makes of Read Across substance 4404-43-7 a conservative representative because of the potential higher bioavailability. Justification for Read Across is detailed in the Toxicokinetics 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

Remarks:

Pre GLP.

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:

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:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wblferstrasse 4, CH-4414 Fullinsdorf.
- Age at study initiation: 10 weeks (males), 12 weeks (females).
- Weight at study initiation: 225 - 247 g (males), 200 - 222 g (females).
- Housing: individually in Makrolon type-2 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: pelleted standard Kliba 343, Batches 77/90 and 78/90 rat maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst); ad libitum.
- Water: community tap water from Itingen; ad libitum.
- Acclimation period: one week.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 40-70 %
- Air changes: 10 - 15 air changes per hr.
- Photoperiod: 12 / 12 hrs dark / hrs light.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: backs of the animals.
- Preparation: clipping 24 h before application.
- % coverage: 10% of the total body surface.

REMOVAL OF TEST SUBSTANCE
- Washing: with lukewarm tap water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 4 ml
- Concentration: 100%
- For solids, paste formed: yes

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: observations four times during test day 1, and daily during days 2 - 15; weighing on test days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: gross pathology.

Statistics:

The LOGIT-Model could not be applied to the observed rate of death. The toxicity was estimated without use of a statistical model.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality.

Clinical signs:

other: No systemic signs were observed in the animals during the entire observation period. Skin observations: males/females: scales (back) (10/10); general erythema (back) (3/10); males: erythema focal (back) (3/5). All animals had recovered from the local sign

Gross pathology:

No findings noted.

Interpretation of results:

not classified

Remarks:

Migrated information according to he CLP Regulation Criteria used for interpretation of results: EU

Conclusions:

LD50 > 2000 mg/kg bw

Executive summary:

Method

Upon an acute oral single administration (2000 mg/kg bw) and a 15 day post-treatment observation period, the dermal LD50 was determined for the test substance, according to the OECD guideline 402.

Results

LD50 > 2000 mg/kg bw

No deaths accorred and no systemic signs were observed in the animals during the entire observation period. Skin reactions are all recovered after 7 observation days.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

The acute oral toxicity of the test compound in rats of both sexes was tested at the single concentration of 5000 mg/kg bw and observed over a period of 14 days. No deaths occurred, thus the LD50 was determined as higher than 5000 mg/Kg bw (Ciba-Geigy, 1981).

A good GLP Klimisch 1 study for acute dermal toxicity is reported. Upon an acute oral single administration (2000 mg/kg bw) and a 15 day post-treatment observation period, the dermal LD50 was determined for the test substance, according to the OECD guideline 402. No deaths occurred and no systemic signs were observed in the animals during the entire observation period (RCC, Research & Consulting Company AG., 1991).

Furthermore, a summary reporting some toxicological test results is available on CAS 16470-24-9 (main component: 68 %; ramaining composition: sodium chloride, sodium sulphate and water); unfortunately the original study reports cannot be more consulted. The available information included in the toxicological test results summary indicate that the substance is not acutely harmful for both oral and dermal route: an oral LD50 of 11800 mg/kg and a dermal LD5 of 2000 mg/kg were indicated (Sandoz, 1972).

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 has been performed. The result of a test performed on an analogous substance at the maximum allowed concentration of 1890 mg/m3has been reported, where no effect is indicated. The analogous substance, part of the Stilbene Fluorescent Whitening Agents, group 3, is the acid form of the disulphonated derivative dihydroxyethyl derivative, therefore contains the same organic functional groups, but due to the sulphonation/salification degree is less soluble. This property makes of Read Across substance 4404-43-7 a conservative representative because of the potential higher bioavailability (further details are given in the Category Justification Report, Section 13 of the technical dossier).

As a conclusion it can be stated that the substance is not acutely toxic for all the three exposure ways.

REFERENCE

Sandoz (1972). Toxicologische Prüfung eines optischen Aufhellers. Unpublished data. Testing laboratory: Sandoz Agroforschung. Owner company: Archroma GmbH. Report N. 31/72. Report date: 1972-08-28

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. Furthermore the active ingredient tested was 83 %.

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).