Pentasodium 7-(4-(4-(5-amino-4-sulfonato-2-(4-((2-(sulfonato-ethoxy)sulfonyl)phenylazo)phenylamino)-6-chloro-1,3,5-triazin-2-yl)amino-2-ureidophenylazo)naphtalene-1,3,6-trisulfonate

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:

18 March 1996 - 07 June 1996

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 40549/A
Batch No: TV1
Expiration date: 01 Jan 2001
Stability in water: for atleast 48 hours
Solubility in water: approximately 90 mg/L at 20 degree celcius.
Aggregate state under storage conditions: solid (powder)
Colour: yellow brown.
Storage conditions: at room temperature
Safety precautions: Routine hygenic procudures are sufficient to assure personnel health and safety.

Analytical monitoring:

yes

Details on sampling:

The temperature, pH-values and dissolved oxygen concentrations in the test medium of 100 mg test substance/l and the control were measured before the start of the test and once every day during the test. At the same times the behaviour of the test substance in the test medium was observed.

Vehicle:

no

Details on test solutions:

The only concentration tested was nominal 100 mg test substance/L. Additionally, a control was tested in parallel (test water without addition of the test substance). Thus, a limit test was performed in accordance with the test guidelines to demonstrate that the test substance has no toxic effect on the test fish up to this concentration.

This test concentration was based on the results of a range-finding test.

The test medium with the test substance concentration of nominal 100 mg/L was prepared by dissolving the test substance homogeneously in test water by intense stirring. The test medium was freshly prepared just before the start of the test (= introduction of the fish).

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The study was performed with Rainbow trout (Oncorhynchus mykiss Walbaum 1792). The test fish were obtained from Forellenhof Fredelsloh, D-37186 Moringen. In accordance with the test guidelines they were held for more than 3 weeks in the laboratories of RCC and were acclimatized for the last 7 days to the test water and temperature without any medication. During acclimation until one day before the start of the test the fish were fed daily with a commercial trout diet (Kronen-Fisch Aminostart, Alleinfutter für Forellenbrut, Kraftfiitterwerk Wesel GmbH, D-46483 Wesel). During the last 3 weeks prior to the test the mortality in the test fish batch was lower than 2 % of the population. The mean body length of the fish at the start of the test was 4.6 ± 0.3 cm (Mean ± SD), range 4.0 - 5.1 cm, the mean body wet weight 0.6 ± 0.1 g (Mean ± SD), range 0.4 - 0.7 g.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

2.5 mmol/l (= 250 mg/l) as CaCO3

Test temperature:

14.6 - 15.8 °C

pH:

7.6 - 8.0

Dissolved oxygen:

8.9 - 9.6 mg/L

Salinity:

Alkalinity: 0.8 mmol/L

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: The analytically determined test concentrations in the test medium of nominal 100 mg/L ranged from 89 % to 94 % of the nominal value (calculated as the average over all measurements).

Details on test conditions:

One glass-aquarium (length x width x height: 0.54 x 0.29 x 0.395 m) with 40 liter test medium was used for the test concentration and the control. The test media were slightly aerated during the test period to keep the oxygen concentration sufficiently high. At the start of the test 7 fish were introduced into each aquarium in a random order (by random numbers). The fish were not fed during the test.

Due to the results of a pre-test the test substance FAT 40'549/A degrades into a defined reaction product during the performance of the biological test. Since a toxic effect can be due to the main compound as well as to a reaction product a static test was performed to test the effect of the main compound as well as the reaction product.

Test duration: 96 hours
Water temperature: 14.6 - 15.8 °C during the test period
Light: a 16-hour light to 8-hour darkness photoperiod with a 30 minute transition period between light and darkness. Light intensity at light period: approximately 10 - 50 Lux.
Test water: reconstituted water: in deionized water with a conductivity lower than 10 µS/cm analytical grade salts were added and the water was aerated to saturation.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: main compound and degradation product

Basis for effect:

mortality (fish)

Details on results:

In the control and in the test concentration of nominal 100 mg test substance/L all fish survived until the end of the test and no symptoms of intoxication were observed.

Reported statistics and error estimates:

The NOEC (No Observed Effect Concentration) and the LC 0 (concentration without mortality) were determined directly from the raw data. The LOEC (Lowest Observed Effect Concentration), the LLC (Lowest Lethal Concentration), the LC50 and the LC100 could not be determined due to the low toxicity of the test substance.

Sublethal observations / clinical signs:

The test medium of 100 mg test substance/L was intensively coloured by the test substance.

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 was established to be greater than 100 mg/L.

Executive summary:

The acute toxicity of the test substance FAT 40'549/A to Rainbow trout was determined in a 96-hour static test according to the Commission Directive 92/69/EEC, Annex Part C.1, dated December 29, 1992 and the OECD Guideline for Testing of Chemicals, Section 2, No. 203, adopted July 17, 1992. The test was performed in compliance with Good Laboratory Practice Regulations.

A limit test was performed in accordance with the test guidelines to demonstrate that the test substance has no toxic effect on the test fish up to the concentration of nominal 100 mg test substance/l. Thus, the only concentration tested was nominal 100 mg test substance/l and a control.

The analytically determined test concentrations in the test medium of nominal 100 mg/l ranged from 89 % to 94 % of the nominal value (calculated as the average over all measurements). The test substance FAT 40'549/A degrades into a defined reaction product during the performance of the biological test. During the test period of 96 hours the test substance concentration decreased to 59 % of the nominal value if the quantification is based on the main compound. Since a toxic effect can be due to the main compound as well as to a reaction product the reported results are related to the sum of the main compound and the reaction product, and thus to the nominal concentration of the test substance since the mean measured concentrations were in the range of 89 % to 94 % of nominal.

The 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), respectively the 96-hour LC 0 of FAT 40'549/A to Rainbow trout was determined to be at least 100 mg test substance/l. The NOEC might even be higher than this concentration. The 96-hour LOEC (lowest concentration with toxic effects) and the 96 -hour LC 50 were clearly higher than this concentration. These values could not be quantified, since concentrations in excess of nominal 100 mg test substance/L have not been tested.

Description of key information

The 96-hour LC50 was higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The acute toxicity of the test substance FAT 40'549/A to Rainbow trout was determined in a 96-hour static test according to the Commission Directive 92/69/EEC, Annex Part C.1, dated December 29, 1992 and the OECD Guideline for Testing of Chemicals, Section 2, No. 203, adopted July 17, 1992. The test was performed in compliance with Good Laboratory Practice Regulations.

A limit test was performed in accordance with the test guidelines to demonstrate that the test substance has no toxic effect on the test fish up to the concentration of nominal 100 mg test substance/l. Thus, the only concentration tested was nominal 100 mg test substance/l and a control.

The analytically determined test concentrations in the test medium of nominal 100 mg/l ranged from 89 % to 94 % of the nominal value (calculated as the average over all measurements). The test substance FAT 40'549/A degrades into a defined reaction product during the performance of the biological test. During the test period of 96 hours the test substance concentration decreased to 59 % of the nominal value if the quantification is based on the main compound. Since a toxic effect can be due to the main compound as well as to a reaction product the reported results are related to the sum of the main compound and the reaction product, and thus to the nominal concentration of the test substance since the mean measured concentrations were in the range of 89 % to 94 % of nominal.

The 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), respectively the 96-hour LC 0 of FAT 40'549/A to Rainbow trout was determined to be at least 100 mg test substance/l. The NOEC might even be higher than this concentration. The 96-hour LOEC (lowest concentration with toxic effects) and the 96 -hour LC 50 were clearly higher than this concentration. These values could not be quantified, since concentrations in excess of nominal 100 mg test substance/L have not been tested.