4,11-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:

2004-11-12 - 2004-12-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test performed according to relevant guidelines and compliant to GLP. Test results are plausible and well documented.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss Ordinance relating to Good Laboratory Practice, adopted February 2nd, 2000 [RS 813.016.5], based on OECD GLP 1997 (C(97) 186/Final)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 5,12-dihydro-2,9-dimethylquino[2,3-b]acridine-7,14-dione and 5,12-dihydro-2-methylquino[2,3-b]acridine-7,14-dione and 5,12-dihydroquino[2,3-b]acridine-7,14-dione

EC Number:

909-082-0

Cas Number:

938065-79-3

Molecular formula:

Unspecified

IUPAC Name:

Reaction mass of 5,12-dihydro-2,9-dimethylquino[2,3-b]acridine-7,14-dione and 5,12-dihydro-2-methylquino[2,3-b]acridine-7,14-dione and 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

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Test item: Young daphnids of a clone of the species Daphnia magna Straus
- Origin: A clone of this species was originally supplied by the University of Sheffield/UK in 1992, defined by the supplier as clone 5. Since that time, the clone is bred in the laboratories of RCC in reconstituted water of the quality identical to the water quality used in the tests (in respect to pH, main ions, and total hardness) and under temperature and light conditions identical to those of the tests.
- Age: daphnids used for the test were 6-24 hours old and were not first brood progeny.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Water Hardness 2.5 mmol/L (= 250 mg/L as CaCO3)

Test temperature:

19 - 20 °C during the test period. The test was performed in a temperature-controlled room (room temperature continuously monitored):
Control: 20°C (start), 20°C (end)
Treatment: 19°C (start), 20°C (end)

pH:

Control: 7.9 (start), 8.0 (end)
Treatment: 7.9 (start), 7.9 (end)

Dissolved oxygen:

[mg/L]
Control: 9.0 (start), 7.8 (end)
Treatment: 9.1 (start), 7.7 (end)

Salinity:

not applicable

Nominal and measured concentrations:

Nominal concentration 100 mg/L
Due to poor solubility in water, no analytical determination possible.

Details on test conditions:

- The test medium is identical with ISO Test water (1) as specified in OECD 202
- Feeding: none
- Limit-test according to guideline (poor solubility in water):
The test medium was prepared as follows: Due to the low water solubility of the test item, a supersaturated dispersion of the test item with a loading rate of 100 mg/L was prepared by weighing 50.1 mg of the test item into 500 mL of test water. No auxiliary solvent or emulsifier was used. The test item was mixed into the test water as homogeneously as possible using ultrasonic treatment for 15 minutes and intense stirring. The dispersion was stirred for 96 hours at room temperature in the dark to dissolve a maximum concentration of the test item in the dispersion.
Then, the dispersion of the test item was filtered through a glass microfibre filter (Whatman GF/C, maximum pore size approximately 1.2 µm) just before the start of the test. The undiluted filtrate of the dispersion with the maximum concentration of dissolved test item was used as test medium.
The limit test was based on the results of a range-finding test and on pre-experiments to determine the solubility of the test item in test water (without GLP).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, tested at least once a year with the strain used

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: test item poorly soluble in water

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: test item poorly soluble in water

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: test item poorly soluble in water

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: test item poorly soluble in water

Details on results:

The biological results are based on the loading rate of the test item of 100 mg/L. Analytical measurements of the actual test item concentration in the test medium were not possible.
In the control and in the undiluted filtrate with the loading rate of 100 mg/L, no immobilized test organisms were observed during the test period of 48 hours.
No remarkable observations were made concerning the appearance of the test medium. The test medium was a clear non-coloured solution throughout the whole test duration.

Results with reference substance (positive control):

The latest result of the positive control test with potassium dichromate in 2004 (48-hour EC50: 0.83 mg/L, RCC Study no. 852355) showed that the toxic performance was valid and within the historical range of the RCC laboratory (from 1996 to 2004: 48-hour EC50: 0.55 - 1.1 mg/L).

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). The acutal concentration could not be determined due to the low solubility of the submission substance in water and the salt concentration present in the ISO-test medium.

Executive summary:

The acute toxicity of the test item to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202, Part I (1984). A limit test was performed in accordance with the EU Commission Directive 92/69/EEC to demonstrate that the test item has no toxic effect on the test organisms up to the solubility limit of the test item in test water. Due to the low water solubility of the test item, a dispersion of the test item with a loading rate of 100 mg/L was continuously stirred at room temperature in the dark over 96 hours. Then, the dispersion was filtered. The undiluted filtrate with the maximum concentration of dissolved or very finely dispersed test item was used as the test medium. Additionally, a control was tested in parallel. No analytical determination of the test item concentration in the test medium was performed since the analysis of the test item in the filtrates - after comprehensive trials - proved impossible due to the very low solubility of the test item in test water and the matrix of the test water. In the control and in the undiluted filtrate with the loading rate of 100 mg/L, no immobilized test organisms were observed during the test period of 48 hours. The 48-hour NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours) and the 48-hour EC0 of the test item to Daphnia magna were determined to be at the loading rate of 100 mg/L. The 48-hour NOEC and the 48 hour EC0 might even be higher but concentrations above the solubility limit of the test item in test water (loading rate of 100 mg/L) were not tested according to the EU Commission Directive. The 48-hour EC50 and the 48-hour EC100 were clearly higher than the solubility limit of the test item in test water (loading rate of 100 mg/L) since the test item had no toxic effect on the daphnids up to the highest test item concentration which could be dissolved or very finely dispersed in the test water under the conditions of the test. In conclusion, the test item had no acute toxic effects on Daphnia magna up to its solubility limit in test water under the present conditions of the test.