2,7-Naphthalenedisulfonic acid, 4-amino-5-hydroxy-, coupled with 3-aminophenol, diazotized 5-amino-2-[(4-aminophenyl)amino]benzenesulfonic acid and diazotized benzenamine, sodium salts

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:

November from 05 to 09, 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1984

Principles of method if other than guideline:

Method: SAV: MFI 003 02.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Water samples were taken from the bath at the beginning and at the end of the experiment.

Vehicle:

no

Details on test solutions:

The test substance was directly weighed and dissolved in 200 ml dechlorinated tap water, then dissolved up to 3 liters, which was slowly added to the fish bath.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish.
- Source: West Aquarium, Bad Lauterberg, BRD.
- Age at study initiation: 95 days.
- Length at study initiation: 27.9 - 29.2 mm.
- Weight at study initiation: 0.22 - 0.24 g.
- Feeding frequency during the test: none.

ACCLIMATION
- Acclimation period: 12 days.
- Acclimation conditions: kept in dechlorinated tap water.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

21.2 - 23.2 °C

pH:

7.3 - 8.3

Dissolved oxygen:

85 - 99 %

Nominal and measured concentrations:

Nominal concentrations: 56, 100, 178, 316 and 562 ppm
Average measured concentrations: 49, 85, 163, 285 and 523 ppm

Details on test conditions:

TEST SYSTEM
- Test vessel: 5 liter glass aquaria.
- Fill volume: 2.9 litres (3.0 litres minus 2 x 0.05 litre for analytical control).
- Aeration: slowly aerated.
- No. of organisms per vessel: 10 fish per concentration and water control.
- No. of vessels per concentration: 1.
- No. of vessels per control: 1, carried out with dechlorinated tap water.

OTHER TEST CONDITIONS
- Photoperiod: 12 hrs light/12hrs darkness.
- Light intensity: yellow light TL 16/20 W.

TEST MEDIUM / WATER PARAMETERS
- Diluent: dechlorinated tap water.
- Intervals of water quality measurement: pH, dissolved oxygen, temperature were measured after 3 hours and then once every day (after 24, 48, 72 and 96 hours).

EFFECT PARAMETERS MEASURED:
- Mortality was measured after 3 hours and then once every day (after 24, 48, 72 and 96 hours).
- Calculaton: the LC50-values are calculated with the Geometric mean, Spearman-Kärber- and the Probit-Method.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8.
- Range finding study: it was carried at 10, 30, 100, 300 and 1000 ppm.
- Results used to determine the conditions for the definitive study: LC50 was determined as 130.5 ppm.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

117.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Remarks:

graphically determined

Effect conc.:

116 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No abnormal responses of the fish were observed.
LC0 (96h): 100 ppm
LC100 (96h): 178 ppm

MEASURED CONCENTRATIONS
The tested concentrations resulted to be mainteined at least 80 per cent of the nominal one, over the test period.

Sublethal observations / clinical signs:

The fishtoxicity data are presented in the table below.

Tank  nr	Conc. (mg/l)	Length (mm)	Weight (g)	Measurement	Test duration
					start / 3 h	24 h	48 h	72 h	96 h
1	Blanck control	28.9	0.24	dead/alive	0/10	0/10	0/10	0/10	0/10
				O2 %	85	95	97	97	97
				pH	7.8	8.2	8.36	8.3	8.3
				T (°C)	21.2	21.8	21.7	21.5	22.1
2	56	28.0	0.22	dead/alive	0/10	0/10	0/10	0/10	0/10
				O2 %	87	94	96	92	93
				pH	7.7	8.2	8.2	8.2	8.2
				T (°C)	22.6	22.7	22.7	22.4	23.1
3	100	28.6	0.24	dead/alive	0/10	0/10	0/10	0/10	0/10
				O2 %	87	94	92	93	93
				pH	7.6	8.2	8.2	8.2	8.2
				T (°C)	22.7	22.7	22.7	22.5	23.2
4	178	27.9	0.22	dead/alive	0/10	4/6	7/3	10/0	10/0
				O2 %	87	96	96	97
				pH	7.6	8.2	8.3	8.2
				T (°C)	22.8	22.9	22.9	22.6
5	316	29.2	0.24	dead/alive	0/10	7/10	10/0	10/0
				O2 %	85	97	99
				pH	7.4	8.2	8.2
				T (°C)	22.7	22.7	22.8
6	562	29.4	0.23	dead/alive	0/10	10/0	10/0
				O2 %	88	99
				pH	7.3	8.2
				T (°C)	88.6	22.7

Test concentration

Nominal conc. ppm	Conc. Start		Conc. End		Average conc.
	ppm	% of nominal	ppm	% of nominal
56	48	85.7	49	87.5	49
100	84	84.0	86	86.0	85
178	162	91.0	164	92.1	163
316	283	89.6	286	90.5	285
562	523	93.1	522	92.9	523

Conclusions:

LC50 (96h): ca 117.7 mg/l (meas. geom. mean)

Executive summary:

The toxicity of the substance to fish (Zebrafish) was assessed in a static acute toxicity test according to the EU Method C.1. 10 fish per concentration and water control were used. Fish were exposed to the following concentration, over a period of 96 hours: 56, 100, 178, 316 and 562 ppm. Water samples were taken from the bath at the beginning and at the end of the experiment; the tested concentrations resulted to be mainteined at least 80 per cent of the nominal one, over the test period.

No abnormal responses of the fish were observed. The LC0 at 96 hours resulted to be 100 ppm, while the LC100 resulted to be 178 ppm.

The LC50-values were calculated with the geometric mean, Spearman-Kärber- and the Probit-Method; furthermore, the value has also been graphically determinated. The LC50 at 96 hours resulted to be 117.7 ppm and 116 ppm, based on geometrical mean and graphically determinated, respectively.

Conclusion

LC50 (96h): 117.7 mg/l (meas. geom. mean)

LC50 (96h): 116 mg/l (graphically determined)

Description of key information

Not harmful/toxic for short-term exposure to fish

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

117.7 mg/L

Additional information

The toxicity of the substance to fish was assessed in a static acute toxicity test using Zebrafish, according to the EU Method C.1. 10 fish per concentration were used. Fish were exposed to the following concentration, over a period of 96 hours: 56, 100, 178, 316 and 562 ppm. Water samples were taken from the bath at the beginning and at the end of the experiment and the tested concentrations resulted to be maintained at least 80 % of the nominal one, over the test period. No abnormal responses of the fish were observed. The LC0 at 96 hours resulted to be 100 ppm, while the LC100 resulted to be 178 ppm. The LC50 at 96 hours resulted to be 117.7 ppm and 116 ppm, based on geometrical mean and graphically determined, respectively.

A second test is available. The experiment was performed using five Zebrafish per test concentration, under semi-static conditions. Fish were exposed to three concentrations, i.e. 46, 64 and 100 mg/l. One death occurred at the concentration of 64 mg/l after 24 hours; at the highest concentration of 100 mg/l 3 fish died after 24 hours and the remaining fish died after 48 hours. The LC50 was determined to be 86.4 mg/l of active ingredient.