Amines, C10-14-branched and linear alkyl, bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-) (1:1)

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1989

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C10-14-branched and linear alkyl, bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-) (1:1)

EC Number:

284-628-2

EC Name:

Amines, C10-14-branched and linear alkyl, bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-) (1:1)

Cas Number:

84961-40-0

Molecular formula:

C32H22CrN10O8.C10-14H21-29NH2

IUPAC Name:

Amines, C10-14-branched and linear alkyl, bis[2,4-dihydro-4-[(2-hydroxy-5-nitrophenyl)azo]-5-methyl-2-phenyl-3H-pyrazol-3-onato(2-)]chromate(1-) (1:1)

Test material form:

solid

Details on test material:

- State of aggregation: solid (powdeQ

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No. of test material: Abl. 13-1341

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: At the start of the test (after 0 hours) samples from vessels which were not stocked with daphnids were analyzed and at the end of the test (after 48 h) samples from stocked and unstocked vessels were analyzed.
- Sample storage conditions before analysis: refrigerator

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: For the preparation of the stock solution (100 mg/L) the test substance was stirred for about 20 h at 22 ± 1 °C in a M4-Medium. Undissolved test substance was removed by paper filtration and centrifugation for about 20 min at about 6000 rpm. By dilution of this stock solution with test medium the following nominal concentrations were prepared: 100 mg/L, 50 mg/L, 25 mg/L, 12.5 mg/L, 6.25 mg/L, 3.13 mg/L, 1.56 mg/L, 0.78 mg/L
- Medium: The synthetic medium M4 was used for the culture and test. The medium was prepared on the basis of an ultrapure, deionized water (conductivity < 0.05 µS/cm). The medium has the composition according to the guideline. The medium was aerated until saturated with oxygen, and was left to stand tor 24 hours for stabilization.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUSS
- Source: Institut National de Recherche Chimique Appliquées, France.
- Culture conditions: The daphnids were cultured under standard conditions in the laboratory.
- Feeding during the test: None
- Age at study initiation: 2 – 24 hours
- Age of the stock of animals: 2 – 4 weeks

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

2.20 – 3.20 mmol/L

Test temperature:

19.5 – 21.1 °C

pH:

7.85 - 8.5

Dissolved oxygen:

7.3 – 8.3 mg/L

Salinity:

- Alkalinity up to pH 4.3: 0.80 – 1.00 mmol/L
- Molar ratio Ca:Mg: ~ 4:1

Conductivity:

550 - 650 µS/cm

Nominal and measured concentrations:

- Nominal concentrations: 100, 50, 25, 12.5, 6.25, 3.13, 1.56 and 0.78 mg/L.
- Measured concentrations: The following nominal concentrations of the test substance were analyzed: 100, 12.5, 1.56 and 0 mg/L (control). The analytical recovery rate was after 48 hours in all analyzed test solutions below the detection limit (< 1 %). The effective concentrations mentioned are based therefore on the nominal concentrations of the test substance.

Details on test conditions:

TEST SYSTEM
- Test vessel: Test tubes (glass) with flat bottom (nominal volume 20 mL)
- Test volume: 10 mL
- Aeration: yes
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 0.5 animals/mL


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: For the preparation of the stock solution (100 mg/L) the test substance was stirred for about 20 h at 22 ± 1 °C in M4-Medium. Undissolved test substance was removed by paper filtration and centrifugation for about 20 min at about 6000 rpm.
- Alkalinity up to pH 4.3: 0.80 – 1.00 mmol/L
- Ca/mg ratio: 4:1
- Conductivity: 550 - 650 µS/cm
- Intervals of water quality measurement:
Measurement of temperature: After 0, 24, 48 hours in an extra vessel close to the test vessels.
Measurement of pH und oxygen: After 0 and 48 hours in one replicate at each concentration.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Illumination/Photoperiod: Artificial light, OSRAM L58 W31 warm white, day:night – rhythm 16:8 hours
- Light intensity: About 5 - 6 µE/(m2-s) in the range of 400 – 700 nm

EFFECT PARAMETERS MEASURED: This study investigated the influence of the test substance on the swimming ability (mobility) of Daphnia magna after 48 hours of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Test concentrations:
Nominal concentrations: 100, 50, 25, 12.5, 6.25, 3.13, 1.56 and 0.78 mg/L.
Measured concentrations: The following nominal concentrations of the test substance were analyzed: 100, 12.5, 1.56 and 0 mg/L (control). The analytical recovery rate was after 48 hours in all analyzed test solutions below the detection limit (< 1 %). The effective concentrations mentioned are based therefore on the nominal concentrations of the test substance.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% C.L not determined; no toxic effects within the range of solubility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% C.L not determined; no toxic effects within the range of solubility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% C.L not determined; no toxic effects within the range of solubility

Results with reference substance (positive control):

not determined

Reported statistics and error estimates:

No reported statistics and error were estimated.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test substance is with high probability not acutely harmful to aquatic invertebrates. No toxic effects occurred within the range of solubility.