Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-[(2-cyanoethyl)[2-(2-hydroxyethoxy)ethyl]amino]-1,3,5-triazine-4,2-diyl]imino]]bis[benzene-1,4-disulphonate]

Endpoint summary

Description of key information

Not harmful/toxic if swallowed

Not harmful/toxic by dermal contact

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From March the 17th to April the 07th,1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

Justification for read across is detailed in section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted on 24 February, 1987

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

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: Charles River Wiga, Sulzfeld, West-Germany
- Age at start of treatment: ca. 7 weeks
- Body weight at start of treatment: maales 201 - 213 g; females 148 - 175 g
- Identification: earmark- Diet: free access to standard pelleted laboratory animal diet; certificate of analysis has been created.
- Water: free access to tap-water; certificate of analysis has been created.
- Acclimatisation: at least 5 days under laboratory conditions.
- Accomodation: housed in groups of five per sex in polycarbonate cages containing purified sawdust as bedding material.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: 7.5 - 15 ACH
- Photoperiod: 12 hours dark / day

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

TREATMENT
- Dose volume: 20 ml/kg b.w.
- Dosage preparation: the formulations were prepared immediately prior to dosing. The test substance was weighed into a glass flask on an analytical balance and the vehicle (w/w) was added. Homogeneity of the test substance in vehicle was obtained by a homogeniser.

Doses:

5000 mg/kg b.w.

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of mortality observations: periodic intervals on the day of dosing (day 1) and twice daily thereafter for at least 14 days.
- Frequency of weighing: day 1 (pre-administration), 8 and 15.
- Frequency of symptoms observation: periodic intervals on the day of dosing (day 1) and once daily thereafter (except day 9 and 10).
- Necropsy of survivors performed: yes.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occured.

Clinical signs:

No clinical signs of toxicity or behavioural.

Body weight:

One female lost 3 grams in the second week; all other animals showed body weight gain during the study period.

Gross pathology:

Macroscopic examination of all animals at termination did not reveal any abnormalities.

Individual animal observations

Sex	Dose level [mg/kg b.w.]	Observations	No. Animals showing effect at exposure time of
			Hours				Day
			0.05	2.1	3.45		2	3	4	5	6	7	8	9	10	11	12	13	14	15
Males	5000	Abnormalities	0	0	0		0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Deaths	0	0	0		0	0	0	0	0	0	0	0	0	0	0	0	0	0
Females	5000	Abnormalities	0	0	0		0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Deaths	0	0	0		0	0	0	0	0	0	0	0	0	0	0	0	0	0
n.a.: inadvertently, no clinical observations performed

Individual body weights

Sex	Dose level [mg/kg b.w.]	Animal no.	Body weight [g] on day			Body weight gain [g] at end
			1	8	15
M	5000	1	206	249	267	67
		2	201	242	258	57
		3	213	275	304	91
		4	208	272	293	85
		5	210	274	302	92
Mean			208	262	285	77
s.d.			4.5	15.7	21	16.9
F	5000	6	173	195	212	39
		7	175	189	195	20
		8	149	168	172	23
		9	151	177	180	29
		10	148	162	159	11
Mean			159	178	184	24
s.d.			13.6	13.8	20.5	10.4

Interpretation of results:

other: not classified, according to CLP Regulation (EC) No1272/2008

Conclusions:

LD50 (male and female) > 5000 mg/kg b.w.

Executive summary:

The test item was administered to rats of both sexes by oral gavage at 5000 mg/kg body weight, followed by a 15 day observation period. The study was conducted in accordance with the method and procedures outline into the OECD guideline 401.

No mortality occurred during the study period. No clinical signs of toxicity or behavioural changes were seen during the study. Apart from one female that lost 3 grams in the second week, all animals showed body weight gain during the study period.

Macroscopic examination of all animals at termination did not reveal any abnormalities that were considered to have arisen as a result of treatment.

Conclusion

LD50 (male and female) > 5000 mg/kg b.w.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

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

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From March the 24th to April the 07th,1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Justification for type of information:

Justification for read across is detailed in section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted on 24 February, 1987

Qualifier:

according to guideline

Guideline:

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

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: Charles River Wiga, Sulzfeld, West-Germany
- Age at start of treatment: ca. 6 weeks
- Weight at start of treatment: males 207 - 239 g; females 162 - 196 g
- Diet: free access to standard pelleted laboratory animal diet; certificate of analysis was create.
- Water: free access to tap-water; certificate of analysis was created.
- Accodomation: housed in groups of five per sex in polycarbonate cages containing purified sawdust as bedding material.
- Acclimation period: at least 5 days under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: 7.5 - 15 ACH
- Photoperiod: 12 hours dark daily

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

TEST MATERIAL PREPARATION
The formulations were prepared immediately prior to dosing. The test substance was weighed into a glass flask on an analytical balance and the vehicle (w/w) was added. Homogeneity of the test substance in vehicle was obtained by a homogeniser. Concentration of test substance in vehicle was varied to allow constant dosage volume in terms of ml/kg body weight.

TEST SITE
- Shaving (clipping): one day before treatment; area of ca. 5x7 cm on the back of the animal.
- Application area: 5x5 cm for males; 3.5x5 cm for females.
- Type of wrap: gauze patch fixed to aluminium foil and flexible bandage with drops of petrolatum.

TEST MATERIAL
- Dose level: 2000 mg/kg b.w.
- Vehicle volume applied: 10 ml/kg b.w.
- Removal: by tissue moistoned with tap-water.

Duration of exposure:

24 hours

Doses:

2000 mg/kb b.w.

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of mortality observations: periodic intervals on the day of dosing (day 1) and twice daily thereafter for at least 14 days.
- Frequency of weighing: day 1 (pre-administration), 8 and 15
- Frequency of symptoms observation: periodic intervals on the day of dosing (day 1) and once daily thereafter.
- Frequency of skin irritation observation: described immediately after bandage removal (day 2) and on days 5, 8 and 15.
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No deaths occurred.

Clinical signs:

All animals were noted as lethargic on the day of dosing. There were no other signs of toxicity or behavioural changes seen over the 15 day observation period.
Red spots were noted on the treated skin surface of three animals.

Body weight:

All animals showed body weight gain during the study period.

Gross pathology:

Macroscopic examination of all animals at termination did not reveal any abnormalities.

Individual animal observations

Sex	Dose level [mg/kg b.w.]	Observations	Animal no. Showing effects at time
			Hour				Day
			0.1	2.15	4.3		1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
M	2000	Abnormalities	0	5	5		0	0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Lethargy	0	5	5		0	0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Deaths	0	0	0		0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
F	2000	Abnormalities	0	5	5		0	0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Lethargy	0	5	5		0	0	0	0	0	0	0	0	n.a.	n.a.	0	0	0	0	0
		Crust in the neck	0	0	0		0	0	0	0	0	0	0	1	n.a.	n.a.	0	0	0	0	0
		Deaths	0	0	0		0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
n.a.: inadvertently no clinical signs were recorded

Individual body weights

Sex	Dose level [mg/kg b.w.]	Body weight [g] on day			Body weight gain [g] at end
		1	8	15
M	2000	216	263	306	90
		207	232	267	60
		227	260	296	69
		239	273	311	72
		218	259	292	74
Mean		221	257	294	73
S.d.		12.1	15.2	17.1	10.9
F	2000	162	182	202	40
		196	209	214	18
		189	208	229	40
		178	191	208	30
		174	190	199	25
Mean		180	196	210	31
S.d.		13.2	11.9	11.9	9.6

Incidence of treated skin abnormalities

Dose level [mg/kg b.w.]	Observations	Day of evaluation
		2		5		8		15
		M	F	M	F	M	F	M	F
2000	Abnormalities	2	1	0	0	0	0	0	0
	Animals with red spots	2	1	0	0	0	0	0	0

Interpretation of results:

other: not classified, according to CLP Regulation (EC) No 1272/2008

Conclusions:

LD50 (male and female) > 2000 mg/kg b.w.

Executive summary:

The test article was dermally administered to a group of 5 males and 5 female rats with a single dose of 2000 mg/kg b.w. The study was conducted in accordance with the method and procedures outline into the OECD guideline 402.

No deaths and no clinical signs of toxicity were observed during the observation period, as well as body weight changes. In addition, the macroscopic examination revealed no organ abnormalities.

Conclusion

LD50 (male and female) > 2000 mg/kg b.w.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

ACUTE TOXICITY BY ORAL ROUTE

The acute oral toxicity of Optical Brightener 380 was tested using 5 males and 5 females Wistar rats, which were dosed at 15000 mg/kg bw, by oral gavage. On the basis of a preliminary test outcomes, the main test was conducted at the limit dose of 15000 mg/kg bw. No deaths occurred and no signs of poisoning were observed during the observation period of 2 weeks. The body weight gain was normal and no autopsy findings were observed.

Since, the lot tested was characterized by a limited content of Optical Brightener 380, the available experiment conducted on the structural analogous Similar Substance 01 has been taken into consideration, in order to confirm the study outcomes.

The read across approach can be considered reliable and appropriate to investigate the property (details for the approach are included into the IUCLID section 13).

Similar Substance 01 was administered to rats of both sexes by oral gavage at 5000 mg/kg body weight, followed by a 15 day observation period. The study was conducted in accordance with the method and procedures outline into the OECD guideline 401. No mortality occurred during the study period. No clinical signs of toxicity or behavioural changes were seen during the study. Apart from one female that lost 3 grams in the second week, all animals showed body weight gain during the study period. Macroscopic examination of all animals at termination did not reveal any abnormalities that were considered to have arisen as a result of treatment.

ACUTE TOXICITY - INHALATION ROUTE

No acute toxicity studies by inhalation route are available on Optical Brightener 380. Nevertheless, because of the physical state and the trade forms of the substance under registration, inhalation is not an appropriate route of exposure. Acute toxicity results for the other exposure routes indicate no concern.

ACUTE TOXICITY BY DETRMAL ROUTE

Since, no specific experimental is are available on dermal acute toxicity potential of Optical Brightener 380, the available experiment conducted on the structural analogous Similar Substance 01 has been taken into consideration. The read across approach can be considered reliable and appropriate to investigate the property (details for the approach are included into the IUCLID section 13).

Similar Substance 01 was dermal administered to a group of 5 males and 5 female rats with a single dose of 2000 mg/kg bw. The study was conducted in accordance with the method and procedures outline into the OECD guideline 402. No deaths and no clinical signs of toxicity were observed during the observation period, as well as body weight changes. In addition, the macroscopic examination revealed no organ abnormalities.

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.1 acute toxicity section, acute toxicity means those adverse effects occurring following oral or dermal administration of a single dose of a substance or a mixture, or multiple doses given within 24 hours, or an inhalation exposure of 4 hours. 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 2000 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).

Dermal LD50 value was established to be greater than 2000 mg/kg body weight, therefore the test substance is out of any classification limit for acute dermal toxicity (dermal acute toxicity category 4: 1000 < ATE ≤ 2000 mg/kg bw).

In conclusion, the substance is not classified for oral, nor dermal acute toxicity, according to the CLP Regulation (EC) No 1272/2008.