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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
From 05 NOV 2004 to 11 NOV 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study compliant to GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guidelineopen allclose all
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)
Remarks:
GLP according to German Chemikaliengesetzt and Directive 88/320/EEC

Test material

Constituent 1
Reference substance name:
2-[[1-[[(2,3-dihydro-2-oxo-1H-benzimidazol-5-yl)amino]carbonyl]-2-oxopropyl]azo]benzoic acid
EC Number:
250-830-4
EC Name:
2-[[1-[[(2,3-dihydro-2-oxo-1H-benzimidazol-5-yl)amino]carbonyl]-2-oxopropyl]azo]benzoic acid
Cas Number:
31837-42-0
IUPAC Name:
2-({2-oxo-1-[(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl)carbamoyl]propyl}diazenyl)benzoic acid

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
Stock solution The solution (100 mg/L test item was weighed out) was prepared with dilution water one day prior to application.
Dispersion treatment The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 micro-m (RC, SCHLEICHER & SCHUELL).
Limit concentration Saturated solution (100 mg/L)
Dilution water According to EC 92/69 L383A C.2. Annex
Saturated solution: The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna STRAUS (Clone 5)
Origin Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
Breeder DR.U.NOACK-LABORATORIEN, 31157 Sarstedt, Germany

Culture In 2-3 L glass vessels with approximatively 1.8 L culture medium, at 21 °C (temperatures of 20 - 25 °C are tolerated), in an incubator, 16 h illumination, illumination strength max. 20 µmol photons*mE(-2)*sE(-1).
Culture medium Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
Feeding of the culture At least 5x weekly ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 10E(6) cells/mL.
Age of test organisms 2 - 24 h old

Acclimation period: approx. 2h

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
total hardness of 160 to 180 mg CaCO3/L
Test temperature:
18 - 22 °C, ± 1 °C constant
pH:
Control: 7.6 - 7.85
Treatment: 7.63 - 7.69
Dissolved oxygen:
[mg/L]
Control: 7.87 - 8.66
Treatment: 8.13 - 8.35
Nominal and measured concentrations:
Nominal limit concentation: 100 mg/L
Control: 0 mg/L
Details on test conditions:
Illumination Max. 20 µmol photons*mE(-2)*sE(-1)
Photoperiod 16 / 8 hours light / dark cycle
Exposure vessel type Glass beakers (5 cm ID x 8 cm H), 50 mL volume were used.
Number of study organisms 20 per concentration and control divided into 4 parallel samples
Feeding during the study The daphnids were not fed during the study.
Endpoints assessed Immobilisation of daphnids
Measurements Prior to test start pH-value, dissolved oxygen concentration,conductivity, total hardness and temperature of the dilution water were measured. At the beginning of the test water parameters (pH value, oxygen concentration) were measured in one additional replicate per concentration and control groups. After 48 h the water parameters in old media were measured in all replicates per concentration and control. The room temperature was recorded throughout the test with a thermohygrograph.
Age of test organisms: 2-24 h old

2 to 24 h old daphnids were used for the definitive study. They were obtained by removing the mother animals twice within 22 h.
Control replicates with dilution water (test medium without test item) were tested under the same conditions as the test groups.
20 daphnids divided into 4 (2 for the reference item) groups with 5 (10 for the reference item) animals per test vessel were used.
In the definitive study the immobilisation of the daphnids was observed after 24 and 48 hours of exposure.
The reference item was tested in five concentrations within the concentration range 0.58 to 5.8 mg/L.
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 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: As the test item is highly insoluble in water, a saturated solution was prepared at the nominal concentration and filtrated
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: As the test item is highly insoluble in water, a saturated solution was prepared at the nominal concentration and filtrated
Duration:
24 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: As the test item is highly insoluble in water, a saturated solution was prepared at the nominal concentration and filtrated
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: As the test item is highly insoluble in water, a saturated solution was prepared at the nominal concentration and filtrated
Details on results:
The test solution was clear throughout the test.
Neither in the control nor in the treatment replicates were any immobilzed daphnids observed.
Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC100-value was determined directly from the test results. The EC10- and EC50- with 95 % confidence interval (CI) was determined in a probability network by interpolation according to standard procedures.
EC-values after 24 h of the reference item in mg/L:
EC10 : 0.97
EC50 : 2.07 (CI 1.68 - 2.56)
EC100 : 5.80
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according to AQS: DIN Guideline 38412 L 30
Reported statistics and error estimates:
The effects of the limit concentration were determined directly from the test results.
EC50- and EC10-values were only calculated for the reference item via probit analysis according to WEBER (1986). Calculation of the confidence intervals was carried out using standard procedures according to BREITIG G. & TÜMPLING W. (1982).

Software
All data were computer generated and rounded for presentation from the full derived data. Consequently, if calculated manually based on the given data minor variations may occur from these figures. Calculations were carried out using software: SigmaPlot rel. 2000 (2000), SPSS CORPORATION

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
At the limit concentration of 100 mg/L of the submission substance no effects on mobility to Daphnia magna were observed. Neither in the control nor in the treatment replicates were any daphnids immobilized.
EC0 (24 and 48 h) = 100 mg/L nominal concentration
EC50 (24 and 48 h) > 100 mg/L nominal concentration
Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution (100 mg/L; the maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6) as limit concentration of the test item was determined according to OECD 202 (1999, Draft) and EC Directive 92/69/EC Method C.2 (1992). The limit test was conducted under static conditions over 48 h from. 20 test organisms were exposed to the limit concentration and control. A reference test was carried out with potassium dichromate to determine the toxicity of the reference item. The EC50-value of the reference item of 2.07 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS, DIN Guideline 38412 L 30. Water quality parameters pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

Table 1: Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure (n = 20)

Test
Concentration

IMMOBILISATION [%]

24 h Replicates

48 h Replicates

1.

2.

3.

4.

MV

1.

2.

3.

4.

MV

Saturated
solution
(100 mg/L)

0

0

0

0

0

0

0

0

0

0

Control

0

0

0

0

0

0

0

0

0

0