Reaction mass of trisodium [m[7-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-8-hydroxynaphthalene-1,6-disulphonato(7-)]]dicuprate(3-) and trisodium [m[4-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-3-hydroxynaphthalene-2,7-disulphonato(7-)]]dicuprate(3-)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From October 28th to December 20th, 1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

April 4, 1984

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Concentrations were measured at the beginning and at the end of the experiment.

Vehicle:

no

Details on test solutions:

The test solutions were prepared by adding approximate amounts of a stock solution of the test material to test water, keeping the volume of the test solution at 4 litres in each test vessel.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Name: zebrafish, Brachydanio rerio, Hamilton-Buchanan.
- Source: Stubenvoll, Allschwil, Switzerland.
- Length at study initiation: 3 - 4 cm
- Weight at study initiation: 0.57 g

ACCLIMATION
- Acclimation period: acclimatization for a minimum of 12 days in 100 liter glass tanks.
- Acclimation conditions: areated test water of 23 ± 2 °C, under flow-through conditions.
- Feeding: fed daily, until 24 hours before starting the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

22 - 23 °C

pH:

7.2 - 8.0

Dissolved oxygen:

5.4 - 8.6 mg/l, i.e. 60 - 95 % saturation

Nominal and measured concentrations:

100, 180, 320, 560 and 1000 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass tanks (150 x 200 x 200 mm) with 4 l test solution.
- Aeration: none.
- No. of organisms per concentration: 10 fish.
- No. of vessels per concentration: 2 replicate.
- No. of vessels per control: 1 replicate.
- Biomass loading rate: 5 fish per 4 litres test solution, i.e. 0.71 g fish/l.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: carbon filtered drinking water.
- Total hardness: about 180 mg CaCO3/l.
- pH: 7.6 to 8.0.

OTHER TEST CONDITIONS
- Light: fluorescent lighting.

MEASUREMENTS
Dissolved oxygen, pH and temperature were measured daily in each test vessel.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No deaths occurred over the 96-hours study period at none of the tested concentrations.

Sublethal observations / clinical signs:

Measured concentrations

Nominal concentration (mg/l)	At the beginning of the test, DOC (mg/l)	At the end of the test, DOC (mg/l)	At the end of the test, % of DOC at beginning
Control	3.5	5	-
Test item 100	47	27	50.5
Test item 180	69	58	81
Test item 320	132.5	105	78
Test item 560	225	155	68
Test item 1000	395	305	77

Test conditions

	O2 (mg/l)
	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control	8.6	7.8	7.4	7.6	7.7
Test item 100	8.1	8.2	7.4	5.8	6.1
Test item 180	7.9	8.4	7.3	5.8	5.9
Test item 320	7.5	8.1	7.6	6.7	6.7
Test item 560	7.0	8.1	7.1	7.0	6.9
Test item 1000	5.4	7.7	6.6	6.8	7.1
	% oxygen saturation
	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control	95	86	82	84	84
Test item 100	90	89	80	62	67
Test item 180	88	90	79	63	64
Test item 320	83	88	82	72	73
Test item 560	77	87	76	76	74
Test item 1000	60	82	71	74	77
	pH
	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control	7.2	7.7	7.9	8.0	8.0
Test item 100	7.3	7.8	7.9	7.6	7.8
Test item 180	7.2	7.9	7.9	7.7	7.7
Test item 320	7.3	7.9	8.0	7.9	7.9
Test item 560	7.2	7.8	7.9	7.9	7.9
Test item 1000	7.2	7.8	7.9	7.9	8.0
	Temperature (°C)
	0 hrs	24 hrs	48 hrs	72 hrs	96 hrs
Control	23	23	23	23	23
Test item 100	23	22	22	22	22
Test item 180	23	22	22	22	22
Test item 320	23	22	22	22	22
Test item 560	23	22	22	22	22
Test item 1000	23	22	22	22	22

Conclusions:

LC50 (96h) > 1000 mg/l (nominal, corresponding to ca 770 mg/l, based on DOC measurement).

Executive summary:

The potential for acute toxicity to fish of the test item was investigated in zebrafish (Brachydanio rerio, Hamilton-Buchanan). Fish were tested in a static experiment, conducted over a period of 96 hours. Stability of test item during the experimental period was checked measuring the DOC vessel content, at the beginning and at the end of the experiment. The percentage of the DOC amount at the end of the test respect to the quantity of the beginning was 50.5, 81, 78, 68 and 77 %, corresponding to the nominal cocnetration of 100, 180, 320, 560 and 1000 mg/l, respectively.

No deaths occurred over the 96-hours study period at none of the tested concentrations.

Conclusion

LC50 (96h) > 1000 mg/l (nominal, corresponding to ca 770 mg/l, based on DOC measurement).

Description of key information

Non harmful/toxic for short-term exposures to fish. LC50 (96h) higher than 1000 mg/l (nominal, corresponding to ca 770 mg/l, based on DOC measurement).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

770 mg/L

Additional information

The potential for acute toxicity to fish of the test item was investigated in zebrafish (Brachydanio rerio, Hamilton-Buchanan). Fish were tested in a static experiment, conducted over a period of 96 hours. Stability of test item during the experimental period was checked measuring the DOC vessel content, at the beginning and at the end of the experiment. The percentage of the DOC amount at the end of the test respect to the quantity of the beginning was 50.5, 81, 78, 68 and 77 %, corresponding to the nominal cocnetration of 100, 180, 320, 560 and 1000 mg/l, respectively. No deaths occurred over the 96-hours study period at none of the tested concentrations.