N-[4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]phenyl]acetamide

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

other: EC Directive 92/69/EC Method C.2 (1992)

GLP compliance:

yes

Remarks:

Deviation from GLP principles - at the sponsor’s request no test item analysis was carried out.

Analytical monitoring:

no

Remarks:

At the sponsor’s request no test item analysis was carried out.

Vehicle:

no

Details on test solutions:

The solution (100 mg/L test chemical was weighed out) was prepared with dilution water one day prior to application in the test medium. The solubility of test substance is < 1 mg/L (related to the pigment). The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 μm

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: STRAUS
- 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 μE.m-2.s-1.
- Culture media: Culture medium Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
- Strain/clone: Clone 5
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
- Age of parental stock (mean and range, SD): 2 - 24 h old
- Feeding during test: The daphnids were not fed during the study.


ACCLIMATION
- Acclimation period: Ca. 2 h
- Type and amount of food: The daphnids were fed with Desmodesmus subspicatus and Chlorella vulgaris during the study.
- Food type: Desmodesmus subspicatus and Chlorella vulgaris with an algae cell density of > 106 cells/mL.
- Amount: ad libitum
- Frequency: Atleast 5x weekly
- Health during acclimation (any mortality observed):

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

266 mg CaCO3/L

Test temperature:

18 - 22 °C, ± 1 °C constant

pH:

7.80

Dissolved oxygen:

8.69 mg/L

Conductivity:

658 μS/cm

Nominal and measured concentrations:

Saturated solution (100 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers were used as a test vessel.
- fill volume: 50 ml
- Volume of the study 20 mL medium
- Number of study organisms: 20 animals, divided into 4 (2*) parallel samples, each with 5 (10*) animals
- No. of vessels per concentration (replicates): 4 replicates for test conc.. and 2 replicates for reference substance.
- Illumination : Diffuse light, illumination range max. 20 20 μE.m-2 .s-1
- Photoperiod : 16/8 h light/dark cycle

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: According to EC 92/69 L383A C.2. Annex. Components of the dilution water includes KCl, NaHCO3, CaCl2.2H2O, MgSO4.7H2O

Reference substance (positive control):

yes

Remarks:

potassium dichromate

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: At the saturated solution (100 mg/L) of the test item has no biologically significant effect was determined

Results with reference substance (positive control):

For , referance substance
EC10 : 1.51
EC50 : 1.84 (CI 1.74 - 1.95)
EC100 : 3.20

Reported statistics and error estimates:

EC10 and EC50 values were only calculated for the reference item via probit according to WEBER (1986). Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982).

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

IMMOBILISATION [%]
	24 h					48 h
	Replicates					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
Control	0	0	0	0	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The median effect concentration (EC50) of saturated solution of test chemical was observed to be >100 mg/l after 48 hrs of exposure period.

Executive summary:

Acute Immobilisation Test for 48 h in static methode to Daphnia magna STRAUS was performed according to OECD-Guideline 202 for Testing of Chemicals (2004) EC Directive 92/69/EC Method C.2 (1992). The solution (100 mg/L test chemical was weighed out) was prepared with dilution water one day prior to application in the test medium. The solubility of test substance is < 1 mg/L (related to the pigment). The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 μm .The effect of the saturated solution (100 mg/L) as limit concentration of the test chemical was determined.The limit test was conducted under static conditions over 48 h . 20 test organisms were exposed to the limit concentration and control. No test chemical analysis was carried out. A reference test was carried out with potassium dichromate to determine the toxicity of the reference substance. EC10 and EC50 values were only calculated for the reference chemical via probit according to WEBER (1986). Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982). The median effect concentration (EC50) of saturated solution of test chemical was observed to be >100 mg/l after 48 hrs of exposure period.

Description of key information

Acute Immobilisation Test for 48 h in static method to Daphnia magna STRAUS was performed according to OECD-Guideline 202 for Testing of Chemicals (2004) EC Directive 92/69/EC Method C.2 (1992) (Experimental study report, 2005). The solution (100 mg/L test chemical was weighed out) was prepared with dilution water one day prior to application in the test medium. The solubility of test substance is < 1 mg/L (related to the pigment). The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 μm .The effect of the saturated solution (100 mg/L) as limit concentration of the test chemical was determined. The limit test was conducted under static conditions over 48 h. 20 test organisms were exposed to the limit concentration and control. No test chemical analysis was carried out. A reference test was carried out with potassium dichromate to determine the toxicity of the reference substance. EC10 and EC50 values were only calculated for the reference chemical via probit according to WEBER (1986). Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982). The median effect concentration (EC50) of saturated solution of test chemical was observed to be >100 mg/l after 48 hrs of exposure period.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Acute Immobilisation Test for 48 h in static method to Daphnia magna STRAUS was performed according to OECD-Guideline 202 for Testing of Chemicals (2004) EC Directive 92/69/EC Method C.2 (1992) (Experimental study report, 2005). The solution (100 mg/L test chemical was weighed out) was prepared with dilution water one day prior to application in the test medium. The solubility of test substance is < 1 mg/L (related to the pigment). The stock solution was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL) and membrane filtrated with 0.45 μm .The effect of the saturated solution (100 mg/L) as limit concentration of the test chemical was determined. The limit test was conducted under static conditions over 48 h. 20 test organisms were exposed to the limit concentration and control. No test chemical analysis was carried out. A reference test was carried out with potassium dichromate to determine the toxicity of the reference substance. EC10 and EC50 values were only calculated for the reference chemical via probit according to WEBER (1986). Calculation of the confidence intervals were carried out using standard procedures according to BREITIG & TÜMPLING (1982). The median effect concentration (EC50) of saturated solution of test chemical was observed to be >100 mg/l after 48 hrs of exposure period.