Reaction mass of Cobaltate(2-), [2-[[[4-hydroxy-3-[[2-oxo-1-[(phenylamino)carbonyl]propyl]azo]phenyl]sulfonyl]amino]benzoato(3-)][2-[[2-hydroxy-5-[(phenylamino)sulfonyl]phenyl]azo]-3-oxo-N-phenylbutanamidato(2-)]-, disodium and Cobaltate(3-), bis[2-[[[4-hydroxy-3-[[2-oxo-1-[(phenylamino)carbonyl]propyl]azo]phenyl]sulfonyl]amino]benzoato(3-)]-, trisodium and sodium bis[2-[[2-hydroxy-5-[(phenylamino)sulphonyl]phenyl]azo]-3-oxo-N-phenylbutyramidato(2-)]cobaltate(1-)

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 November the 18th, 1985 to January the 31st, 1986

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 test solutions:

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

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish.
- Source: Stubenvoll, Allschwil, Switzerland.
- Length at study initiation: 2.5 - 4 cm
- Weight at study initiation: 0.69 g

ACCLIMATION
- Acclimation period: acclimatisation for a minimum of 12 days.
- Acclimation conditions: in 100 l glass tanks, in aerated water test water at 23 ± 2 °C, under flow-through conditions.
- Feeding frequency during acclimation: fed until 24 hours before starting the test.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Vessels: 22 - 25 °C
Control: 22 - 24 °C

pH:

Vessels: 7.2 - 8.0
Control: 7.2 - 8.0

Dissolved oxygen:

Vessels: 7.0 - 8.5 mg/l, i.e. 79 - 97 % oxygen saturation
Control: 7.0 - 8.3 mg/l, i.e. 79 - 95 % oxygen saturation

Nominal and measured concentrations:

Nominal 3.2, 5.6, 10, 18, 32 mg/l

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 vessel: 5 fish per 4 l test solution.
- Biomass loading rate: 0.86 g fish / I
- No. of vessels per concentration: 2 replicates.
- No. of vessels per control: 2 replicates.

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: carbon filtered drinking water with a total hardness of about 180 mg CaCO3/l and pH 7.6 to 8.0.

OTHER TEST CONDITIONS
- Light: fluorescent lighting.

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

EFFECT PARAMETERS MEASURED
The cumulative percentage mortality was plotted against the concentration on Logarithmic-probability paper.
A line was fitted, by eye, and the concentration corresponding to the 50 per cent response read off.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The maximum concentration producing no mortality resulted to be 3.2 mg/l
The minimum concentration producing 100 % mortality resulted to be 10 mg/l

MEASURED CONCENTRATIONS
The measured concentrations were in the range of ± 20 % the nominal one, over the entire observation period, except in the case of the nominal concentration of 3.2 mg/l, of which the geometrical mean revelaed that it was ca 79 % of the nominal one.

Sublethal observations / clinical signs:

RATE OF DEATHS

Nominal conc. (mg/l)	No fish	No of deaths after hrs				Mortality % after 96 %
		24 hrs	48 hrs	72hrs	96 hrs
Control	10	0	0	0	0	0
3.2	10	0	0	0	0	0
5.6	10	0	1	1	1	10
10	10	10	10	10	10	100
18	10	10	10	10	10	100
32	10	10	10	10	10	100

MEASURED CONCENTRATIONS

Nominal conc. (mg/l)	At the start (mg/l)	At the end (mg/l)	Geom mean (mg/l)	% of nominal
Control	-	-		-
3.2	2.3	2.8	2.54	79.30
5.6	4.1	5.0	4.53	80.85
10	8.9	9.4	9.15	91.47
18	16.8		16.80	93.33
32	31		31.00	96.88

TEST CONDITIONS

Nominal conc. (mg/l)	0 h	24 hrs	48 hrs	72hrs	96 hrs	0 h	24 hrs	48 hrs	72hrs	96 hrs
	O2 (mg/l)					% oxygen saturation
Control	8.3	7.6	7.4	7.3	7.0	95	82	82	81	79
3.2	8.4	7.6	7.3	7.0	7.1	95	83	82	79	80
5.6	8.3	8.0	7.9	7.5	7.4	95	87	87	84	83
10	8.5	8.2				97	89
18	8.4					96
32	8.4					96
	pH					Temperature (°C)
Control	7.2	7.7	7.9	7.9	8.0	24	22	22	22	22
3.2	7.3	7.7	8.0	8.0	8.0	24	22	23	23	23
5.6	7.2	7.8	8.1	8.1	8.1	25	22	22	23	23
10	7.3	7.6				25	22
18	7.2					25
32	7.2					25

Conclusions:

LC50 (96h): 6 mg/l (estimated, nominal conc.)

Executive summary:

The short-term toxicity to fish potential of the test substance was evaluated under static test conditions, according to the procedures outlined into the OECD guideline 204 and EEC Directive 84/449 C,1 test method. The substance was tested at the nominal concentrations of 3.2, 5.6, 10, 18, 32 mg/l.

The measured concentrations were in the range of ± 20 % the nominal one, over the entire observation period, except in the case of the nominal concentration of 3.2 mg/l, of which the geometrical mean revealed that it was ca 79 % of the nominal one.

The maximum concentration producing no mortality resulted to be 3.2 mg/l, while the minimum concentration producing 100 % mortality resulted to be 10 mg/l.

Conclusion

LC50 (96h): 6 mg/l (estimated, nominal conc.)

Description of key information

LC50 (96h): 6 mg/l (estimated, nominal conc.)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

6 mg/L

Additional information

The short-term toxicity to fish potential of the test substance was evaluated using zebrafish, under static test conditions; the experiment was conducted in accordance with the procedures outlined into the OECD guideline 204 and EEC Directive 84/449 C,1 test method. The substance was tested at the nominal concentrations of 3.2, 5.6, 10, 18, 32 mg/l. The measured concentrations were in the range of ± 20 % the nominal one, over the entire observation period, except in the case of the nominal concentration of 3.2 mg/l, of which the geometrical mean revealed that it was ca 79 % of the nominal one.

The maximum concentration producing no mortality resulted to be 3.2 mg/l, while the minimum concentration producing 100 % mortality resulted to be 10 mg/l. The LC50 (96h) value was estimated to be 6 mg/l (estimated, nominal conc.).

Since the Acid Yellow 235 lot tested in the key study was characterized by an appreciable non-hazardous impurity content, the available data on structural analogous Similar Substance 01 have been taken into account, in order to confirm the study outcomes. The read across approach can be considered as reliable and adequate for the purpose; details and explanations are detailed in the report attached to the IUCLID section 13.2.

An acute fish toxicity test was performed following a internal method, similar to the OECD test guideline 203. Fish were exposed to 2, 20 and 100 mg/l over a period of 48 hours to determine the Tolerate Level values. The TL0 resulted to be 2 mg/l, while the TL100 was identified to be 20 mg/l. therefore, the TL50 (48h) was estimated to be 6.3 mg/l (graphically determined).