5,12-dihydroquinolino[2,3-b]acridine-7,14-dione and (mono and bis)-[(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)methyl]-5,12-dihydroquinolino[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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study according to OECD 202

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of mono-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)methyl]-5,12-dihydroquino[2,3-b]acridine-7,14-dione and di-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)methyl]-5,12-dihydroquino[2,3-b]acridine-7,14-dione and 5,12-dihydroquino[2,3-b]acridine-7,14-dione

EC Number:

700-776-7

Molecular formula:

C29H17O4N3 (unspecified isomers) / C38H22O6N4 (unspecified isomers) / C20H12O2N2

IUPAC Name:

Reaction mass of mono-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)methyl]-5,12-dihydroquino[2,3-b]acridine-7,14-dione and di-[(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)methyl]-5,12-dihydroquino[2,3-b]acridine-7,14-dione and 5,12-dihydroquino[2,3-b]acridine-7,14-dione

Details on test material:

- Physical state: Red powder
- Storage condition of test material: Room temperature, protected from light, in original container.

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The water accommodated fractions (WAF) was prepared with a nominal loading of the test item of 100 mg/L as specified below.
100 mg/L of the test item were weighed out and transferred with an appropriate amount of dilution water into a brown glass flask. This dispersion was shaken for 24 h with 20 rpm at room temperature.
After a separation phase of 1 h, the aqueous phase or WAF was removed by siphoning from the approximate centre of the glass
flask to give the 100 mg/L loading rate WAF to be used as a stock WAF.
The Iimit loading Ievei of 100 mg/L was based on the results of a non GLP preliminary range finding test conducted at the test facility und er static conditions.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Strain: Daphnia magna STRAUS (Clone 5).
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBolu).
- Age at study initiation (mean and range, SD): 2 to 24 h old daphnids were used for the study. They will be obtained by removing the parental daphnids twice within 22 h.
- Culture: ln glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 ± 2°C, in an incubator, 16 h illumination, light intensity of max. 20 ln glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 ± 2°C, in an incubator, 16 h illumination, light intensity of max. 20 µE*m^-2*s^-1. The pH should be in the range of 6 to 9.
- Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
Culture feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata, Desmodesmus
subspicatus and Chlorella vulgaris, with an algae cell density of > 10^6 cells/m L.
- Feeding during test: None

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

160 to 180 mg CaCO3/L.

Test temperature:

18 - 22°C, ± 1 °C constant

pH:

7.88 - 8.08

Dissolved oxygen:

7.55-8.67 mg/L

Nominal and measured concentrations:

100 mg/L (nominal, WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 ml capacity.
- Aeration: No
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Same as for culture.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16/8 h Iight/dark cycle.
- Light intensity: Diffuse light, light intensity of max. 20 Diffuse light, light intensity of max. 20 µE*m^-2*s^-1
- Prior to the start of the exposure (0 h), the water quality parameters (i.e. pH value, dissolved oxygen concentration, temperature, conductivity and total hardness) of the dilution water were measured. ln the fresh media (0 h), the water quality parameters (i.e. pH values, dissolved oxygen concentrations) were measured in one additional replicate of the WAF and the control. ln the old media (48 h), the water quality parameters were measured in all replicates of the WAF and the control. The room temperature was recorded throughout the test period.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobilisation was determined in all groups after 24 and 48 h. An animal is considered to be immobile, if it is not able to swim within 15 sec after gentle agitation of the test vessel.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: The Iimit loading Ievei of 100 mg/L was based on the results of a non GLP preliminary range finding test conducted at the test facility under static conditions.
- Range finding study: Yes
- Test concentrations: 100 mg/L WAF (No analytical evaluation of the Iimit concentration and the control was carried out. The solubility of the test item in water turned out to be lower than the Iimit of quantification of the analytical system of 0.2 mg test item/L.)

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate (WAF)

Remarks on result:

other: No acute toxic effects within the range of solubility.

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate (WAF)

Remarks on result:

other: No acute toxic effects within the range of solubility.

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate (WAF)

Remarks on result:

other: No acute toxic effects within the range of solubility.

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EL10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: loading rate (WAF)

Remarks on result:

other: No acute toxic effects within the range of solubility.

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC50-value with 95 % confidence interval (Cl) was calculated by sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50 : 1.92 mg/L (CI 1.74- 2.10 mg/L)
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 P 9/2 (05/1996) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

With high probability acutely not harmful to aquatic invertebrates. No acute toxic effects within the range of solubility.