Dichloro-5,12-dihydroquino[2,3-b]acridine-7,14-dione

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

Study period:

2005-03-18 - 2005-04-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study had been performed according to relevant guidelines and compliant to GLP. The results are plausible and well documented.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

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

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP according to German Chemikaliengesetz and directive 88/320/EEC

Test material

Test material information

Constituent 1

Reference substance name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

EC Number:

213-879-2

EC Name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

Cas Number:

1047-16-1

IUPAC Name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
not applicable

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

not applicable

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 µm (RC, SCHLEICHER & SCHUELL).
Limit concentration: Saturated solution (100 mg/L)

CONTROL: Dilution water without test item tested under the same conditions as the test groups.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna STRAUS (Clone 5), bred in the test facility.

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 µE/(m2 ¿ s).

Culture medium: Culture medium Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.

Feeding: At least 5x weekly ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 106 cells/mL.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

not applicable

Test conditions

Hardness:

Total hardness of 266 mg CaCO3/L

Test temperature:

20 +/- 1°C

pH:

Start:
control: 7.8 -- treatment: 7.7

End:
control: 7.83 - 7.94 -- treatment: 7.4 - 7.8

Dissolved oxygen:

[mg/L]
Start:
control: 8.69 -- treatment: 8.87

End:
control: 7.76 - 7.93 -- treatment: 7.91 - 7.98

Salinity:

not applicable

Nominal and measured concentrations:

Due to the poor solubility of the submission substance in water a limit test at 100 mg/L nominal concentration had been performed.

Details on test conditions:

A static preliminary range finding test was conducted as limit test with the saturated solution (NON-GLP-state).
The definitive study was performed as a limit test with saturated solution (100 mg/L) under static conditions to enable the determination of immobilisation after 24 and 48 hours. 2 to 24 h old daphnids were used for the definitive study. They were obtained by removing the mother animals twice within 22 h. The daphnids were acclimated to the dilution water ca. 2 hours before test start. The daphnids were not fed during the study.
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*) groups with 5 (10*) animals per test vessel were used.
In the definitive study the immobilisation of the daphnids was observed after 24 and 48 hours of exposure. Water parameter were measured at begin and end of the study. The reference item was tested in five concentrations within the concentration range 0.58 to 5.8 mg/L.
*) The information in brackets refer to the test of the reference item

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Due to poor solubility of test item in water, limit test at nominal concentration of 100 mg/L

Effect concentrations 2

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: Due to poor solubility of test item in water, limit test at nominal concentration of 100 mg/L

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Due to poor solubility of test item in water, limit test at nominal concentration of 100 mg/L

Effect concentrations 4

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: Due to poor solubility of test item in water, limit test at nominal concentration of 100 mg/L

Details on results:

The percentage immobility was determined for the tested limit concentration and the control after 24 h and 48 h. In both, the control and treatment replicates, no immobilization occured . There is no biologically significant effect neither in the tested limit concentration nor in the control group.

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: 1.51
EC50: 1.84 (CI 1.74-1.95)
EC100: 3.2
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.

AQS-Merkblatt: Bestimmung der nicht akut giftigen Wirkung von Abwasser gegenüber Daphnien über Verdünnungsstufen (DIN 38412 - L 30)

Reported statistics and error estimates:

not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In a 48 hour chronic toxicity test (reliability category 1, compliant to GLP) on Daphnia manga Straus performed as a static limit test due to the low solubility of the submission substance, the NOEC (48 h) had been determined to be the nominal concentration of 100 mg/L. The EC50 is higher than the nominal limit concentration (100 mg/L).

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS) the effect of the saturated solution (100 mg/L) as limit concentration of the test item, was determined according to OECD 202 (2004) and EC Directive 92/69/EC Method C.2 (1992). The limit test was conducted under static conditions over 48 h from 2005-03-22 to 2005-03-24 at DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Straße 1, D-31157 Sarstedt. 20 test organisms were exposed to the limit concentration and control. No test item analysis was carried out. 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 1.84 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.

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) Control	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0	0 0

MV = Mean value