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]

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted on 24 February, 1987

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Similar Substance 01

IUPAC Name:

Similar Substance 01

Test animals

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

Administration / exposure

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

Results and discussion

Effect levels

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.

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.