2-Naphthalenesulfonic acid, 7-amino-4-hydroxy-8-[2-[2-sulfo-4-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]diazenyl]-, potassium sodium salt (1:?:?), coupled with diazotized 2-[(4-amino-5-methoxy-2-methylphenyl)sulfonyl]ethyl hydrogen sulfate

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:

Study initiation date - 28 May 2003; Experiment start date - 02 June 2003; Experiment end date - 21 June 2003; Study completion date - 30 July 2003.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40812/A
Batch: WP 8/03
Purity: approx. 75 %
Appearance: Solid, dark red-brownish powder
Expiration date: 23 April 2010
Storage: At room temperature at about 20 °C

Analytical monitoring:

yes

Details on sampling:

Specimens of each test concentration were drawn from the approximate centre of the test vessels. They were taken at the beginning and the end of the exposure period.

Vehicle:

no

Details on test solutions:

Stock solution: 299.9 mg test item were weighed, dissolved, and made up to 3 litres with test water.
Test concentration: Since no mortality was observed in the range-finding test with the nominal test concentration of 100 mg/L, the definitive test was performed as limit test with the nominal test item concentration of 100 mg/L. Additionally a blank control was carried out.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Origin: Commercial supplier: Institut für Hirnforschung, Universität Zürich-Irchel, 8057 Zürich, Switzerland. Fish supplied in one batch were all of a similar age and size.
- Holding: Fish were held in well water of "Lange Erlen" and "Hard" and fed daily (except for weekends) with a commercially prepared zebrafish food diet supplemented by living organisms such as daphnia or artemia. Fish used in the test were not fed 96 hours prior to initiation of the test. Quality of the fish was assessed during an acclimatisation period of at least 12 days in well water. Fish were not treated for disease during holding.
- Total length: (mean) 28 mm ± 2 mm (range 24-30 mm) based on 7 fish of the control.
- Body weight: (mean) 0.15 g ± 0.04 g (range 0.11-0.22 g) based on 7 fish of the control
- Loading: Fish biomass to solution ratio was 0.34 g/L.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

160 mg/L CaCO3

Test temperature:

22.3 - 23.4 °C

pH:

7.8 - 8.3

Dissolved oxygen:

96 - 101 % of the saturation value

Nominal and measured concentrations:

- Nominal: 100 mg/L
- Measured: ranging from 103% to 110% of nominal between start and end of exposure

Details on test conditions:

Experimental conditions:
- Lighting: Fluorescent light, 16 hours daily with 30 min transition period
- Test vessels: 5 litre glass aquaria with a test volume of 3 litre, slightly aerated
- Replicates: Performed with one test chamber per concentration and control
- Test organisms: 7 fish per concentration and control
- Loading: Fish biomass to solution ratio was 0.34 g/L W
- Feeding: Fish were not fed during the test period.

Stock solution and test concentration:
- Stock solution: The stock solution was prepared as follows: 299.9 mg test item were weighed, dissolved, and made up to 3 litres with test water.
- Test concentration Since no mortality was observed in the range-finding test with the nominal test concentration of 100 mg/L, the definitive test was performed as limit test with the nominal test item concentration of 100 mg/L. Additionally a blank control was carried out.

Test water:
Well water of "Lange Erlen" and "Hard" was used as test dilution water. Prior to its use, the water was passed through an activated carbon filter. Water hardness was 160 mg/L CaC03. The chlorine content was under the detection limit (< 0.1 ppm). For more information about the test water.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality of fish was recorded in the control and the nominal test concentration of 100 mg/L FAT 40812/A during the 96-hour exposure period. No sublethal effects such as changes in swimming behaviour, respiratory function, pigmentation, exophthalmos or loss of equilibrium were observed in the control or the treatment concentration of 100 mg/L during the 96 hour test period.

Reported statistics and error estimates:

Due to the absence of mortality during the exposure, the median lethal concentration (LC50) was estimated without statistical program.

Validity criteria fulfilled:

yes

Conclusions:

The 96-h LC50 is >100 mg/L in zebrafish

Executive summary:

The acute toxicity of the test substance to Zebra fish was determined in a GLP-compliant 96 hours static limit test according to OECD Guideline 203. The nominal concentration for the test was 100 mg/L. During the test, the test article concentrations in the analysed test media were in the range of 103 - 110 % of the nominal values indicating that the test substance was sufficiently stable. In the control and the test concentration no mortalities and no sublethal effects such as changes in swimming behaviour, respiratory function, pigmentation, exophthalmos, or loss of equilibrium were observed after 96 hours. The LC50 was higher than 100 mg/L.

Description of key information

The acute toxicity of the test substance to Zebra fish was determined in a GLP-compliant 96-hours static limit test according to OECD Guideline 203. The nominal concentration for the test was 100 mg/L. During the test, the test article concentrations in the analysed test media were in the range of 103 - 110 % of the nominal values indicating that the test substance was sufficiently stable. In the control and the test concentration no mortalities and no sublethal effects such as changes in swimming behaviour, respiratory function, pigmentation, exophthalmos, or loss of equilibrium were observed after 96 hours. The LC50 was higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L