Reaction mass of Sodium 2-(2 or 3-{[(chloroacetyl)amino]methyl}-4-{[4-(cyclohexylamino)-9,10-dioxo-9,10-dihydroanthracen-1-yl]amino}phenoxy)-5-methylbenzenesulfonate and Sodium 2-(3 or 2-{[(chloroacetyl)amino]methyl}-4-{[4-(cyclohexylamino)-9,10-dioxo-9,10-dihydroanthracen-1-yl]amino}phenoxy)-5-methylbenzenesulfonate

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:

22 December, 1993 to 09 March, 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

None

Analytical monitoring:

yes

Details on sampling:

Composite samples of each test concentration were drawn by mixing identical volumes of the test solutions taken from the approximate centre of the test vessels. They were taken immediately before exposure and after 48 hrs exposure and kept at 18 to 22 °C until analysis.

Vehicle:

yes

Details on test solutions:

Stock solution:
1.00 g test substance were mixed with 9.01 g water, by means of a marble mill, during 24 hours. 2.00 g of this blend were mixed and made up to 2000 ml with water.

Preparation of test solutions:
Calculated amounts of the stock solution to produce the desired test concentrations were given into the water and were homogeneously distributed. After an equilibration time of 7 hours the daphnia were transferred into the beakers. The test substance was homogeneously distributed in the test vessels at all test concentrations at the begin of the test. At the end of exposure a deposit was observed at all test concentrations. Values are based on measured concentrations at 0 h.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus 1820
- Source: CIBA-GEIGY Ltd., testing facility
- Breeding: Cultures of daphnia are maintained in glass vessels containing approx. 2.5 L of reconstituted water (see below) at 20±1 °C (water is renewed partially thrice weekly). At each renewal the daphnia are fed with a suspension of green algae (Scenedesmus subspicatus) supplemented by a suspension of TETRAMIN-extract in such quantities that the food is consumed after 24h.
- Pretreatment: 24 hours before the begin of the exposure reproductive daphnia are separated from the young by sieving all individuals through a 800 pm sieve.
This operation is repeated 6 hours before the exposure and the young (6-24h of age) are retained for the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

None

Post exposure observation period:

Not available

Hardness:

240 mg CaC03/L

Test temperature:

20±1 °C

pH:

7.8 - 8.0

Dissolved oxygen:

≥8.8 mg/L

Salinity:

Not available

Nominal and measured concentrations:

Nominal: 4.4, 9.6, 21, 46 and 100 mg test substance/L
Actual : 3.9, 7.9, 18.1, 35.8 and 78.1 mg test substance/L

Details on test conditions:

Type of vessel: Beakers covered with watch glasses 100 ml solution per beaker
Test procedure: static
Duration: 48 hours
Temperature: 20.5 – 21.0 °C
Oxygen concentration: ≥ 8.8 mg/L
pH-value: 7.23 – 7.95
Exposure to light: 16 hours photoperiod / 8 hours darkness daily
Feeding: None during the test
Test vessels: four 100 mL glass beakers per concentration, each filled with 50 mL, one additional replicate for measurements without test organisms
Loading: 10 mL of test solution for each animal
Number of animals: 20 per concentration in 4 replicates of 5

Observations: Immobilizations were recorded after 24 and 48 hours exposure.

Measurements: The oxygen content, pH and temperature were measured at 0 and 48 hours.

Calculâtions/Statistical Analysis: None

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 78 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

None

Results with reference substance (positive control):

Not available

Reported statistics and error estimates:

None

The rate immobilisations observed with test concentrations 0, 3.6, 7.4, 16.3, 34.6 and 75.2 mg/L were 5 %, 5 %, 0 %, 10 %, 5 % and 0 %, respectively.

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h) for immobilisation was determined to be >78 mg/L. The corresponding NOEC (48 h) was 78 mg/L.

Executive summary:

An acute toxicity test on Daphnia magna (Daphnia magna Straus 1820) was performed on FAT 20077/C according to the 92/69/EEC C.2 Acute toxicity for Daphnia method.

In a static assay, 20 daphnia’s were exposed to nominal concentrations of 4.4, 9.6, 21, 46 and 100 mg/L. The corresponding measured concentions were determined to be 0, 3.6, 7.4, 16.3, 34.6 and 75.2 mg/L.

The percentage immobilisations observed with test concentrations 0, 3.6, 7.4, 16.3, 34.6 and 75.2 mg/L were 5, 5, 0, 10, 5 and 0, respectively.

Based on these findings, the following results were obtained:

EC50 (48 h) calculated: >78 mg/l

NOEC (48 h): 78 mg/l

EC0 (48 h): >78 mg/l

EC100 (48h): >78 mg/l

Description of key information

An acute toxicity test on Daphnia magna (Daphnia magna Straus 1820) was performed on FAT 20077/C according to the 92/69/EEC C.2 Acute toxicity for Daphnia method.

In a static assay, 20 daphnia’s were exposed to nominal concentrations of4.4, 9.6, 21, 46 and 100 mg/L.The corresponding measured concentions were determined to be 0, 3.6, 7.4, 16.3, 34.6 and 75.2 mg/L.

The percentage immobilisations observed with test concentrations 0, 3.6, 7.4, 16.3, 34.6 and 75.2 mg/L were 5, 5, 0, 10, 5 and 0, respectively.

Based on these findings, the following results were obtained:

EC50 (48 h) calculated: >78 mg/l

NOEC (48 h): 78 mg/l

EC0 (48 h): >78 mg/l

EC100 (48h): >78 mg/l

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

78 mg/L