Sodium 1-amino-4-(cyclohexylamino)-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From February 07 to February 09, 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

Limit test: 3 concentrations tested

Principles of method if other than guideline:

Internal method: SAV 14 000 03

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 45, 100 and 220 mg test article/L.
- Sampling method: the concentrations of the test substance was calculated at the beginning and at the end of the test using a spectrophotometric method.
- Sample storage conditions before analysis: all test medium samples for the analytical determinations were kept at -18°C to 25°C until analysis.

Vehicle:

yes

Details on test solutions:

- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Tween 80 (Polyoxyethylensorbitanmonooleat).
- Concentration of vehicle in test medium (stock solution and final test solution): 5 ppm.
- Evidence of undissolved material: precipitate.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia Magna.
- Method of breeding: suspension of green algae (Scenedesmus subspicatus) supplemented by a TETRAMIN-extract in such quantites that the food was consumed within 24h.
- Source: Product Ecology TF, Textile Dyes Division.
- Breeding: cultures of Daphnia magna are held in glass vessels containing approx. 3.0 litre of reconstituted water at 18 - 22°C. The water was renewed partially three times per week.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

214 mg/L CaCo3

Test temperature:

21°C

pH:

The pH was in a range of 7.8 - 8.2.
7.8 (in the highest test concentration)

Dissolved oxygen:

Saturated

Nominal and measured concentrations:

Nominal concentration: 45 mg/L, 100 mg/L, 220 mg/L - Measured concentration: 21 mg/L, 37 mg/L, 67 mg/L

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 10
- Type of vessel: 250 ml beakers
- Content of vessel: 100-200 ml test medium
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- Light exposure: 12 hours light/12 hours dark

TEST MEDIUM
The test medium used had the following composition:
NaHCO3 = 65 mg/L
MgSO4*7H2O = 123.3 mg/L
KCl = 5.8 mg/L
CaCl2*2H2O = 294 mg/L
Na2SiC = 4.3 mg/L
NaNO3 = 274 µg/L
KH2PO4 = 143 µg/L
K2HPO4 = 184 µg/L

The test medium was prepared with bidistilled water.
The medium was oxygen saturated at the beginning of the test.

Ca/Mg ratio = 4:1
Na/K ratio = 10:1
alkalinity = 08 mmol/L

STOCK SOLUTION
0.5 g test substance were mixed with 200 ml test medium in a beaker, was homogenized for 10 minutes by ultrasonic treatment and filled up to 500 ml with test medium. Since the test substance was poorly soluble in the test medium, most of the test substance settled down and built a sediment at the bottom of the flask.

TEST CONCENTRATIONS
The nominal test concentrations were chosen as 45/100 and 220 mg test article/I. For the preparation of the test concentrations only the supernatant of the stock solution was taken.

SAMPLING FOR ANALYSIS
All test medium samples for the analytical determinations were taken from the approximate center of the test vessels (without mixing the medium) and kept at -18°C to -25°C until analysis.
The concentrations of the test substance at the beginning and at the end of the test were analysed by the PE Analytical Services K, Textile Dyes Division.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

ca. 67 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

ca. 34 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 67 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 67 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 67 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 67 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Basis for effect:

mobility

Results with reference substance (positive control):

EC50: 0.95 mg/L

Because of the low solubility of the test article in the test media the actual test concentrations in the analysed test media were in the range of 30 - 47% of the nominal values. Therefore, the reported results are related to the actual mean values of the analysed concentrations of the test article measured at the beginning and at the end of the test. No immobilization and abnormal behaviour of the Daphnia was observed in the controls. The NOEC and ECQ value after 48 hours were determined to be 34 mg/L, the EC₅₀ and the EC₁₀₀ value could not be calculated, but are clearly higher than 67 mg/L.

Validity criteria fulfilled:

yes

Conclusions:

EC0 = 67 mg/L
EC50 > 67 mg/L
EC100 > 67 mg/L

Executive summary:

Method:

The short term toxicity to aquatic invertebrates was determined using the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).

 

Observations:

The test concentrations were prepared from a 1.0 g/L stock solution. Although the test substance is soluble in pure (deionised) water ( about 80 g/L), most of the test substance in the stock solution settled down and build a sediment at the bottom of the flask. This effect was possibly caused by a reaction of the test substance with the water hardness compounds in the test water. For the preparation of the test concentrations only the supernatant of the stock solution was taken (limit of the solubility). The nominal concentrations tested were 45/100 and 220 mg test article/L, a vehicle and a blank control without any addtions. In all test concentrations a sediment at the bottom of the test vessel was observed after 48 hours. Because of the low solubility of the test article in the test media, the actual test concentrations in the analysed test media were in the range of 30 - 47% of the nominal values. Therefore, the reported results are related to the actual mean values of the analysed conentrations of the test article measured at the beginning and at the end of the test. No immobilization and abnormal behaviour of the Daphnia was observed in the controls. The NOEC value after 48 hours was determined to be 34 mg/L, the EC₅₀ and the EC₁₀₀ value could not be calculated, but are clearly higher than 67 mg/L.

 

Results:

EC₀ = 67 mg/L

EC₅₀ > 67 mg/L

EC₁₀₀ > 67 mg/L

Description of key information

EC50 (48h) > 67 mg/L (measured)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

67 mg/L

Additional information

The short term toxicity to aquatic invertebrates was determined using the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test).

Although the test substance is soluble in pure (deionised) water ( about 80 g/L), most of the test substance in the stock solution settled down and build a sediment at the bottom of the flask. This effect was possibly caused by a reaction of the test substance with the water hardness compounds in the test water. For the preparation of the test concentrations only the supernatant of the stock solution was taken (limit of the solubility). The reported results are related to the actual mean values of the analysed conentrations of the test article measured at the beginning and at the end of the test. No immobilization and abnormal behaviour of the Daphnia was observed in the controls. The NOEC value after 48 hours was determined to be 34 mg/L, the EC₅₀ and the EC₁₀₀ value could not be calculated, but are clearly higher than 67 mg/L.

Read-Across

The short-term toxicity to aquatic invertebrates was predicted by using the Read Across technique. A similar substance to the test substance was evaluated due to the lack of data for this endpoint. The complete Read-Across justification is attached at section 13.