C.I. Pigment Red 282

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-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:

21 July 1999 to 20 September 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with OECD Guideline 211

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

5,12-dihydro-2,9-dimethylquino[2,3-b]acridine-7,14-dione

EC Number:

213-561-3

EC Name:

5,12-dihydro-2,9-dimethylquino[2,3-b]acridine-7,14-dione

Cas Number:

980-26-7

Molecular formula:

C22H16N2O2

IUPAC Name:

2,9-dimethyl-5,12-dihydroquino[2,3-b]acridine-7,14-dione

Sampling and analysis

Analytical monitoring:

no

Test solutions

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock dispersion was prepared for each test interval by adding 5 mg of the test substance into a 250 mL beaker. Approximately 150 mL water was added and the mixture was homogenized with an ultra-turrax and by ultrasonification (30 minutes). The mixture was than transferred into a graduated flask, filled up to a volume of 5 L with water for dilution(= 1 mg test substance / L) and stirred for 48 hours with a magnetic stirrer. After a sedimentation phase of 1 hour 100 mL portions of the test dispersion were collected from the middle of the stock dispersion vessel using a glass pipette and subsequently transferred to the test vessels.
The test vessels were pre-conditioned with test solution to prevent adsorption of the test substance which perhaps would reduce the test concentration below the solubility limit. The test vessels were filled with test solution (nominal concentration 1 mg/L), gently swirled and then the test solution was discarded.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: clone 5
- Source: Stock culture of Hoechst Marion Roussel Deutschland GmbH, ProTox
- Age of parental stock (mean and range, SD): daphnids were used for breeding up to an age of 42 days, daphnids with an age < 24 hours were used for the studies
- Feeding during test
- Food type: monocellular green algae (Scenedesmus subspicatus)
- Amount: 0.2 mg carbon / day / animal
- Frequency: three times a week during breeding and daily including weekends during the study.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Test conditions

Hardness:

2.1-2.3 mmol Ca2+ + Mg2+/L (concentration group and control)

Test temperature:

20.8-21.2 °C (concentration group and control)

pH:

8.0-8.2 (concentration group), 7.9-8.2 (control)

Dissolved oxygen:

oxygen content: 9.0-9.4 (concentration group), 8.8-9.3 (control) mg/L

Nominal and measured concentrations:

Nominal: 1 mg/L
Measured: no analytical monitoring (see above)
From the properties of the test substance it could be concluded that the test substance was chemically stable in water over the test intervals of 48 and 72 hours, respectively. The concentration values given in this report are based on the nominal concentration. Due to the very low solubility of the substance it can be estimated, that the actual concentration is ¿ 20 µg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL beakers (height 90 mm, diameter 50 mm), filled with 70 mL test water (according to Elendt, M4) served as culture vessels, 150 mL beakers, each filled with 100 mL test water, as test chambers
- Renewal rate of test solution (frequency/flow rate): the daphnids were transferred into fresh prepared test solutions on Monday, Wednesday and Friday
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted fresh water was used as water for breeding and dilution. Demineralized water with a conductance of < 1µS / cm was used for preparing the artificial fresh water (according to Elendt M4).
- The test water was allowed to stand to stabilise for at least a further 24 hours and was aerated until saturated with oxygen.
- Intervals of water quality measurement: The pH-value, oxygen content, conductivity and total hardness were measured and recorded once a week.

OTHER TEST CONDITIONS
- Photoperiod: 16 h/d (fluorescent tubes)
- Light intensity: 300 Ix

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Every 24 hours: immobility, reproduction rate (number of living and dead offspring) and development of embryos in the brood pouch

VEHICLE CONTROL PERFORMED: no vehicle

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 1 mg/L

Nominal / measured:

nominal

Basis for effect:

immobilisation

Effect concentrations 2

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 1 mg/L

Nominal / measured:

nominal

Basis for effect:

reproduction

Effect concentrations 3

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.02 mg/L

Conc. based on:

other: solubility limit: ¿ 0.02 mg/L

Basis for effect:

immobilisation

Effect concentrations 4

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.02 mg/L

Conc. based on:

other: solubility limit: ¿ 0.02 mg/L

Basis for effect:

reproduction

Details on results:

No immobility occurred in the 1 mg/L (nominal concentration) and in the control group.
No differences in the onset of brood production were observed in the concentration group in comparison to the control. The reproduction rate in the concentration group showed no statistically significant changes in comparison to the control (see table below for details).

Reported statistics and error estimates:

On the premises of a monotonous concentration-effect-relationship the Wilcoxon-Test was applied

Any other information on results incl. tables

	control	1 mg/L
		C.I. Pigment Red 122
n	10	10
mean of living offspring	102.2	109.6
standard deviation	15.61	18.56
coefficient of variation	15.3	16.9
median	98.5	106.0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

The mortality of the parent animals did not exceed 20 % at the end of the test in the controls and the mean number of live offspring produced per parent animal surviving at the end of the test was >=60

Conclusions:

In this prolonged toxicity study with Col. Pigment Red 122 in Daphnia magna over 21 days the following effect concentrations were obtained:
NOEC (immobility):
> 1 mg/L (nominal concentration), > solubility limit ¿ 20 µg/L)
NOEC (reproduction rate):
> 1 mg/L (nominal concentration), > solubility limit ¿ 20 µg/L)
No adverse effects of C.I. Pigment Red 122 on Daphnia magna were observed at the maximum tested concentration of 1 mg/L (nominal concentration) which was above the solubility limit (¿ 20 µg/L) of the test substance in the test water.

Executive summary:

C.l. Pigment Red 122 was tested in Daphnia magna (water flea) over 21 days in a semistatic system. Since no effects were expected up to the limit of solubility a limit test with a nominal concentration of 1 mg/L was conducted.

A negative control (0 mg/L) and a nominal concentration of 1 mg/L were tested. The tested concentration is already above the limit of solubility of the test substance in water (¿ 20 µg/L).

The test was conducted under semistatic conditions. The media and the test solutions were renewed every Monday, Wednesday and Friday. For every renewal new test solutions were prepared. This was done by stirring the test substance for 48 hours into the test medium. The aqueous phase of the concentration group was present as a slight reddish clear solution during the entire exposure period. There were no visible particulate matters. Water parameters were within the normal range.

Analytical monitoring of the concentration of the test substance in the test water was not conducted since no relevant results could be expected. The solubility limit of the test substance is below the analytical detection limit. From the properties of the test substance it could be concluded that the test substance is chemically stable in water over the test intervals of 48 and 72 hours, respectively.

The concentration values given in this report are based on the nominal concentration.

In the control group no mortality occurred and the mean number of living offspring produced per parent animal was 102.2, thus fulfilling the validity criteria for the test.

No immobility occurred in the 1 mg/L (nominal concentration) and in the control group. No difference in the onset of brood production was observed in the concentration group in comparison to the control. The reproduction rate in the concentration group showed no statistically significant changes in comparison to the control.

In this prolonged toxicity study with C.I. Pigment Red 122 in Daphnia magna over 21 days the following effect concentrations were obtained:

NOEC (immobility): > 1 mg/L (nominal concentration), > solubility limit (¿ 20 µg/L)

NOEC (reproduction rate): > 1 mg/L (nominal concentration), > solubility limit (¿ 20 µg/L)