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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

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Diss Factsheets

Administrative data

short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Meets the criteria for classification as Reliable without restriction according to Klimisch et al (1997)

Data source

Reference Type:
study report

Materials and methods

Test guideline
according to guideline
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
C35H31N8O18S3.xNa.yK, where x+y=3
dipotassium sodium 3-[(1E)-2-{4-[(1E)-2-{4-[(1E)-2-(5-carbamoyl-1-ethyl-2-hydroxy-4-methyl-6-oxo-1,6-dihydropyridin-3-yl)diazen-1-yl]-2-sulfonatophenyl}diazen-1-yl]-2,5-bis(2-hydroxyethoxy)phenyl}diazen-1-yl]-4,5-dihydroxynaphthalene-2,7-disulfonate

Sampling and analysis

Analytical monitoring:
Details on sampling:
The concentrations of test substance in the test solutions were measured at 0 and 96 hours using the high performance liquid chromatography.

Test solutions

Details on test solutions:
This study was run with a single nominal concentration of 120 mg/l of test substance together with a dilution water control. The test solution was prepared by the addition of a known amount of test substance directly to 20 litres of dilution water in the test vessel, resulting in a dark blue/black solution.

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
The test organism was the rainbow trout Oncorhynchus mykiss. The fish (batch 04/04) were obtained from Houghton Springs Fish Farm, Winterbourne Houghton, Blandford Forum, Dorset, DT11 OPD, UK on 20 January 2004 and held in stock tanks at Brixham Environmental Laboratory for at least 12 days before the start of the test. Prior to the test the fish were acc1imatised to the test temperature (15 ± 1°C) for a minimum period of 7 days.

The last medication given to the fish was a 10 day course of amoxycyllin. The treatment was completed on 9 February 2004, 14 days prior to the test. No mortality was observed in the batch of fish in the 7 days prior to the test.

During the holding and acc1imatisation periods the fish were fed appropriate amounts of a commercial fish food. Food was withheld from the fish for at least 24 hours prior to the commencement of the test.

At the end of the exposure period, the fish from the dilution water control were weighed and measured. The range in weight was 1.31 to 2.60 g with a mean of 1.73 g, and the range in length was 45 to 54 mm with a mean of 48 mm.

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
96 h

Test conditions

The total hardness of the dilution water was 49.3 mg/l CaC03, and the conductivity of the water was 228 uS cm-I @ 25°C.
Test temperature:
Test was undertaken in a temperature controlled room. Test temperature was recorded continuously was within the range 15 ± 1°C.
pH values were recorded at 0, 24, 48, 72 and 96 hours and ranged from 7.3 to 7.7
Dissolved oxygen:
At no time during the course of the study was dissolved oxygen concentration in any of the test vessels less than 60% of the air-saturation value as given in the: National Institute of Oceanography of Great Britain and UNESCO (1973). International Oceanographic Tables, Vol 2
Not applicable - freshwater test
Nominal and measured concentrations:
The overall mean measured concentration was 100% of the nominal value. The percentage loss in the measured concentration over the test period was <1%.

The nominal concentrations were used for the calculation and reporting of the results.
Details on test conditions:
The test procedure employed was a static system. Single borosilicate glass vessels (external dimensions; 400 mm x 280 mm x 280 mm; length x width x height) were used for the dilution water control and the exposure solution. The vessels had a working volume of 20 litres.

The test was undertaken in a temperature controlled room which was set at the nominal test temperature of 15 ± 1°C. The test solutions were gently aerated. The photoperiod in this study was 16 hours fluorescent light and 8 hours dark with 20 minute dawn and dusk transition periods commencing at 06:00 and 21:40 hours. At the start of the test ten fish were randomly allocated to the single test concentration and the dilution water control. The fish were not fed during the course of the test.
Reference substance (positive control):

Results and discussion

Effect concentrationsopen allclose all
96 h
Dose descriptor:
Effect conc.:
> 120 mg/L
Basis for effect:
mortality (fish)
96 h
Dose descriptor:
Effect conc.:
120 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:
The test substance showed no toxicity to fish under the test conditions.