Reaction products of disodium 3,5-diamino-2-[(2-sulfonatophenyl)diazenyl]benzoate and  diazotised sodium 2-[(4-aminophenyl)sulfonyl]ethyl sulfate

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:

Study initiation date - 20 January 2003; Experiment start date - 21 January 2003; Experiment completion date - 14 February 2003; Study completion date - 14 April 2003.

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 40810/A
Batch: WP 6/02
Purity: approx. 75 %
Appearance: Solid, dark brownish powder
Expiration date: 12 December 2010
Storage: At room temperature at about 20 °C

Analytical monitoring:

yes

Details on sampling:

Samples of aqueous solutions of the blank and all test concentrations were taken from the test solutions at the start and at the end of the exposure period. All test medium samples for the analytical determinations were taken from the approximate centre of the test vessels and kept at -18 °C to -25 °C until analysis.

Vehicle:

no

Details on test solutions:

100.1 mg test item were mixed with test medium, treated with ultrasound for one minute and then made up to 1000 ml with test medium in a volumetric flask. The appearance of the solution was red.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test species: Daphnia magna Straus, clone 5
Breeding: Cultures of Daphnia magna were bred in the laboratories of the test facility in glass vessels containing approximately 3 litre of reconstituted water (M4 medium, Elendt 1990). The water was renewed partially twice a week. The Daphnia were fed daily with the exception of the weekend with a suspension of green algae (Scenedesmus obliquus) in such quantities that the food was consumed within 24 hours. Culturing was performed at 18 – 22 °C and a photoperiod of 16 hours light and 8 hours dark. Twice a week the young Daphnia were separated from the parent animals to maintain a stock of reproducing Daphnia within the same age.
Quality control: The quality of the Daphnia is checked at least every 3 months by determining the EC50 value (24 h) for the reference item potassium dichromate. Reference control on December 04, 2002: EC50 = 0.9 mg/L The EC50 for potassium dichromate after 24 hours of exposure was determined among 46 laboratories in an international ring-test with Daphnia magna (European Communities, 1992) and found to be at 1.5 mg/L (0.9 to 1.9 mg/L). Therefore, the cultures of Daphnia magna used by the test facility were demonstrated to be adequately sensitive.
Pre-treatment: About 24 hours before the start of exposure reproductive Daphnia were separated from the young Daphnia by sieving all individuals through a 1000 µm sieve. This procedure was repeated before the start of the exposure, to receive young Daphnia with an age of less than 24 hours.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

231 mg CaCO3/L

Test temperature:

20 °C ± 2 °C, constant throughout the test within ± 1 °C. During the exposure period the water temperature ranged between 19.2 - 20.2 °C in the test vessels.

pH:

The pH ranged from 7.8 to 8.0

Dissolved oxygen:

>60 % of air saturation value. The dissolved oxygen content was between 96 % and 99 % during the test.

Salinity:

Alkalinity 0.8 mmol/L

Nominal and measured concentrations:

A range finding test was performed with concentrations of 0.01, 0.1, 1.0, 10 and 100 mg/L.
The main test was performed with nominal test item concentrations of 4.3, 9.4, 21, 45 and 100 mg/L and a blank control without test item. Measured test concentrations in the analysed samples ranged from 105 to 114 % of the nominal values at the start of the test and from 107 to 116 % of the nominal values at the end of the exposure period.

Details on test conditions:

Test vessels were 250 ml beakers with 100 ml test medium. 20 Daphnia per concentration were used, two beakers per test concentration with 10 animals each. A 16 hours light daily, 8 hours dark cycle was maintained.

TEST SYSTEM
- Test vessel: Beaker
- Material, size, headspace, fill volume: Glass, 250 mL containing 100 mL of test medium.
- Aeration: Yes
- No. of organisms per vessel: 20 Daphnia per concentration
- No. of vessels per concentration (replicates): Two
- No. of vessels per control (replicates): 20 Daphnia per concentration
- No. of vessels per vehicle control (replicates): Two

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light daily, 8 hours dark.

EFFECT PARAMETERS MEASURED: Immobilization

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
- Test concentrations: 0.01, 0.1, 1.0, 10 and 100 mg/L.

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:

The test was considered valid since:
- not more than 10 % of the daphnids in the controls were immobilised and
- the oxygen concentration at the end of the test was >60 % of the air saturation value.

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

No statistical analysis of the data was performed.

After 48 hours of exposure, no immobilisation was observed in the test concentrations of 4.3, 9.4, 21 and 45 mg/L. At the highest test concentration of 100 mg/L, 15 % immobilisation of daphnids had occured after 48 hours of the test.

Validity criteria fulfilled:

yes

Conclusions:

The EC100 and EC50 were both >100 mg/L in this test and the EC0 was set to 45 mg/L. Thus, FAT 40810/A is not considered toxic to daphnia magna.

Executive summary:

The objective of this study was the estimation of the acute toxicity of FAT 40810/A to the Daphnia magna in a 48 hour static test. This test was conducted in accordance with OECD test guideline 202 and EEC guideline C.2 in a GLP-certified laboratory. The nominal concentrations of FAT 40810/A were 4.3, 9.4, 21, 45 and 100 mg/L and a blank control without test item. Test samples of the blank control and the test concentrations taken at the start and at the end of the 48-hour period were analysed for FAT 40810/A. Measured test concentrations in the analysed samples ranged from 105 to 114 % of the nominal values at the start of the test and from 107 to 116 % of the nominal values at the end of the exposure period. Hence, the following results are based on nominal test item concentrations. No immobilisation of daphnids was observed in the control and at any of the test item concentrations after 24 hours. After 48 hours of exposure, no immobilisation was observed in the blank control and test concentrations of 4.3, 9.4, 21 and 45 mg/L. At the highest test concentration of 100 mg/L, 15 % immobilisation of daphnids had occurred after 48 hours of the test.

Description of key information

EC100 and EC50 were both >100 mg/L in an acute daphnia toxicity test with FAT40810/A and the EC0 was set to 45 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The objective of this study was the estimation of the acute toxicity of FAT 40810/A to the Daphnia magna in a 48 hour static test. This test was conducted in accordance with OECD test guideline 202 and EEC guideline C.2 in a GLP-certified laboratory. The nominal concentrations of FAT 40810/A were 4.3, 9.4, 21, 45 and 100 mg/L and a blank control without test item. Test samples of the blank control and the test concentrations taken at the start and at the end of the 48-hour period were analysed for FAT 40810/A. Measured test concentrations in the analysed samples ranged from 105 to 114 % of the nominal values at the start of the test and from 107 to 116 % of the nominal values at the end of the exposure period. Hence the following results are based on nominal test item concentrations. No immobilisation of daphnids was observed in the control and at any of the test item concentrations after 24 hours. After 48 hours of exposure, no immobilisation was observed in the blank control and test concentrations of 4.3, 9.4, 21 and 45 mg/L. At the highest test concentration of 100 mg/L, 15 % immobilisation of daphnids had occurred after 48 hours of the test.