2-{4-[4-[4-fluoro-6-(2-(2-vinylsulfonylethoxy)ethylamino)-1,3,5-triazin-2-ylamino]phenylazo]phenylazo}naphthalene-4,6,8-trisulfonate, trisodium salt

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:

26 March 2001 to 08 May 2001

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)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Analytical monitoring:

yes

Details on sampling:

Samples of each concentration were taken from the test solutions at the start and at the end of exposure period. Additionally, a sample of the stock solution was taken at the start. All test medium samples for the analytical determinations were taken from the approximate centre of the test vessels (after mixing the medium) and kept at -18 °C to - 25 °C until analysis.

Details on test solutions:

STOCK SOLUTION:
100.1 mg test item were mixed with test medium, homogenised by ultrasonic treatment for 1 minute, and then made up to 1000 mL with test medium in a volumetric flask. The appearance of the solution was red.

TEST CONCENTRATIONS:
A range finding test was performed with concentrations of 0.01, 0.1, 1.0, 10 and 100mg/L. based on the pre-test, 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. Calculated amounts of the stock solution to produce the desired test concentrations were given into the test vessels and then made up to 200 mL with test medium. The 200 mL of test solutions were divided up into portions of 100 mL for each replicate test vessel.

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 three times a week. The daphnids were fed daily with the exception of the weekend with a suspension of green algae (Scenedesmus subspicatus) supplemented by a Tetramine extract 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. Three times a week the young daphnids were separated from the parent animals to maintain a stock of reproducing daphnia within the same age.
- Quality control: The quality of the daphnids is checked at least every 3 months by determining the EC50 value (24 h) for the reference item potassium dichromate. Reference control on January 31, 2001: EC50 0.91 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 daphnids were separated from the young daphnids by sieving all individuals
through a 1000 µm sieve. This procedure was repeated before the start of the exposure, to receive young daphnids with an age of less than 24 hours.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

214 mg/L CaC03

Test temperature:

20.1-21.0°C

pH:

8.1 to 8.5

Dissolved oxygen:

The oxygen content was in the range of 96 - 98 %.

Nominal and measured concentrations:

Nominal concentrations: 4.3, 9.4, 21, 45, and 100 mg/L.
Mean measured concentrations: 2.6, 6.3, 15, 38, and 86 mg/L.

Details on test conditions:

- Light: 16 hours light daily, 8 hours dark
- Test vessels: 250 mL beakers containing 100 mL test medium
- Daphnia per concentration: 20, two beakers per test concentration with 10 animals each.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

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 EC-values are based on the mean measured concentrations of FAT 40800/A and the nominal concentrations. No immobilisation of daphnids was observed in the control and at any of test item concentrations. After 48 hours of exposure no sublethal effects on the daphnids were recorded.

Results with reference substance (positive control):

EC50 0.91 mg/L

Reported statistics and error estimates:

no statistics were used

Validity criteria fulfilled:

yes

Conclusions:

The EC50-value is based on mean measured concentrations of FAT 40800/A >86 mg/L. The EC50 used for the assessment is the nominal value, measured is with in 80 % of nominal and in line with the other studies performed for this substance.

Executive summary:

A GLP-compliant Daphnia immobilisation test was carried out according to the OECD 202 and EEC C.2 guideline. Daphnia magna Straus species was used in the test (20 daphnids/test concentration). The water temperature was maintained at 20.1-21.0 °C throughout the exposure period in all test vessels. The oxygen content was in the range of 96 - 98 % during the test and the pH ranged from 8.1 to 8.5. The nominal concentrations were 4.3, 9.4, 21, 45, and 100 mg/L and a blank control without test item. The test samples from the start and after 48 hours of exposure were analysed. The measured test concentrations in the analysed samples were in the range of 60.5 - 83.3 % of the nominal values at the start and 60.5 - 89.6 % at the end of exposure. The mean measured test item concentrations were determined as 2.6, 6.3, 15, 38, and 86 mg/L. The EC-values are based on the nominal concentration. No immobilisation of daphnids was observed in the control and at any of test item concentrations. After 48 hours of exposure no sub-lethal effects on the daphnids were recorded.

Description of key information

The 48-h EC50 value of the test substance is >100 mg/L in Daphnia magna.	

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

> 100 mg/L

Additional information

The acute toxicity of the test substance to aquatic invertebrates was determined in a GLP-compliant test according to OECD 202. In this study, Daphnia magna (10 per test beaker) were exposed to nominal concentrations of 0, 4.3, 9.4, 21, 45, and 100 mg/L of the test substance for 48 hours (static). The measured test concentrations in the analysed samples were in the range of 60.5-83.3 % of the nominal values at the start and 60.5-89.6 % at the end of exposure. The mean measured test item concentrations were determined as 2.6, 6.3, 15, 38, and 86 mg/L. The EC-values are based on the nominal concentration as measured is within 80% of nominal and in line with the other studies performed for this substance. No immobilization or other abnormal behaviour of the Daphnia was observed in any of the test concentrations. The 48 h EC50 with measured concentration was >86 mg/L and therefore, the 48-h EC50 was determined at >100 mg/L.