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 aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 March 1996 to 07 June 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'549/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:

At the start and at the end of the test, the pH-values and the dissolved oxygen concentrations were determined in the test media of all test concentrations and the control. The water temperature was determined in the test medium and in one control beaker at the start and the end of the test.

Vehicle:

no

Details on test solutions:

The test medium of the highest test substance concentration was prepared by dissolving the test substance in test water (100 mg/L). This test medium was diluted in a series of sequential dilutions with test water to prepare the following final nominal concentrations: 4.6, 10, 21, 46 and 100 mg test substance/L. Additionally, a control was tested in parallel (test water without addition of the test substance). The test solutions were prepared just before introduction of Daphnia (= start of the test).

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed with young Daphnia (at the start of the test 6-24 hours old) of a clone of the species Daphnia magna Straus. The clone is bred in the laboratories of RCC under standardized conditions. The cultivation of the parental Daphnia was performed in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 mg/L as CaCO3

Test temperature:

19.9 - 20.7 °C

pH:

7.8 - 8.0

Dissolved oxygen:

8.9 - 9.8 mg/L

Nominal and measured concentrations:

The nominal concentrations tested were 4.6, 10, 21, 46 and 100 mg test substance/l, and in parallel a control.

The analytically determined test concentrations in the test media ranged from 92 % to 96 % of the nominal values (calculated as the average over all measurements per test concentration). The test substance FAT 40'549/A degrades into a defined reaction product during the performance of the biological test. The mean measured concentrations were determined to be 55 % to 62 % of the nominal values, 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 concentrations of the test substance since the mean measured concentrations were well above 90 % of nominal.

Details on test conditions:

In each test concentration and the control 20 Daphnia were tested, divided into four groups of five animals, each group in 60 mL test medium in a glass-beaker (100 ml). The test was performed in an air-temperated room.

Water temperature: 19.9 - 20.7 °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 540 - 800 Lux.

Test duration: 48 hours

Test water: reconstituted water: in deionized water with a conductivity lower than 0.1 µS/cm (Milli-Q-water) analytical grade salts were added, and the water was aerated to oxygen saturation.

The Daphnia have not been fed, and the test media have not been aerated during the test period.

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

In the control and up to and including the test concentration of nominal 100 mg test substance/L no significant rate of immobility or mortality of the test animals was observed during the test period of 48 hours.

In the control and at the test substance concentration of nominal 46 mg test substance/L one Daphnia was immobilized until the end of the test. This rate of immobilization was not determined as a significant toxic effect, because according to the test guidelines this immobility rate is also tolerated in the control, and additionally in the highest test substance concentration of nominal 100 mg test substance/L no immobilization and no other signs of intoxication were observed.

Therefore, the 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), and the 48-hour EC 0 of FAT 40'549/A to Daphnia magna were determined to be at least 100 mg test substance/L. The NOEC might even be higher than this concentration. The 48-hour LOEC (lowest concentration with toxic effects) and the 48-hour EC50 were clearly higher this test concentration. These values could not be
quantified, since concentrations in excess of nominal 100 mg test substance/L have not been tested.

During the test period, the dissolved oxygen concentrations in the test media were at least 8.9 mg/L or higher, the pH-values ranged from pH 7.8 to pH 8.0. All test media were coloured by the test substance.

Reported statistics and error estimates:

The immobility or mortality of the Daphnia was determined by visual controls after 24 and 48 hours. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile.

The NOEC (No Observed Effect Concentration) and the EC0 (highest test concentration without effect) were determined directly from the raw data. The LOEC (Lowest Observed Effect Concentration), the EC50 and the EC100 could not be quantified due to the low toxicity of the test substance.

Validity criteria fulfilled:

yes

Conclusions:

In conclusions, the 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), and the 48-hour EC0 of FAT 40'549/A to Daphnia magna were determined to be at least 100 mg test substance/L. The NOEC might even be higher than this concentration. The 48-hour LOEC (lowest concentration with toxic effects) and the 48-hour EC 50 were clearly higher this test concentration. These values could not be quantified, since concentrations in excess of nominal 100 mg test substance/L have not been tested.

Executive summary:

The acute toxicity of the test substance FAT 40549/A to Daphnia magna was determined in a 48-hour static test according to the Commission Directive 92/69/EEC, Annex Part C.2, dated December 29, 1992, and the OECD Guideline for Testing of Chemicals, Section 2, No. 202, Part I, adopted April 04, 1984. The test was performed in compliance with Good Laboratory Practice Regulations.

The nominal concentrations tested were 4.6, 10, 21, 46 and 100 mg test substance/L, and in parallel a control.

The analytically determined test concentrations in the test media ranged from 92 % to 96 % of the nominal values (calculated as the average over all measurements per test concentration). The test substance FAT 40549/A degrades into a defined reaction product during the performance of the biological test. The mean measured concentrations were determined to be 55 % to 62 % of the nominal values, 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 concentrations of the test substance since the mean measured concentrations were well above 90 % of nominal.

The 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), respectively the 48-hour EC 0 of FAT 40549/A to Daphnia magna was determined to be at least 100 mg test substance/L. The NOEC might even be higher than this concentration. The 48-hour LOEC (lowest concentration with toxic effects) and the 48-hour EC 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 48-hour EC50 was higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The acute toxicity of the test substance FAT 40549/A to Daphnia magna was determined in a 48-hour static test according to the Commission Directive 92/69/EEC, Annex Part C.2, dated December 29, 1992, and the OECD Guideline for Testing of Chemicals, Section 2, No. 202, Part I, adopted April 04, 1984. The test was performed in compliance with Good Laboratory Practice Regulations.

The nominal concentrations tested were 4.6, 10, 21, 46 and 100 mg test substance/L, and in parallel a control.

The analytically determined test concentrations in the test media ranged from 92 % to 96 % of the nominal values (calculated as the average over all measurements per test concentration). The test substance FAT 40549/A degrades into a defined reaction product during the performance of the biological test. The mean measured concentrations were determined to be 55 % to 62 % of the nominal values, 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 concentrations of the test substance since the mean measured concentrations were well above 90 % of nominal.

The 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), respectively the 48-hour EC 0 of FAT 40549/A to Daphnia magna was determined to be at least 100 mg test substance/L. The NOEC might even be higher than this concentration. The 48-hour LOEC (lowest concentration with toxic effects) and the 48-hour EC 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.