1-hydroxy-4-(p-toluidino)anthraquinone

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Purity: 95.2 %
- Batch Number: AAFE 046203
- Chemical name: Solvent Violet 13
- Appearance: Violet powder
- Water solubility: < 1 mg/L
- Stability in water and light at room temperature: Stable
- pH vlaue: Neutral (in aqueous solution)
- Melting point: 185 °C
- TOC: 76.57 %
- Expiry date: 2016-10-27

Analytical monitoring:

yes

Remarks:

The saturated solution and the control after O h (new media) and 48 h (old media) were analytically verified via DOC analysis according to DIN EN 1484.

Details on sampling:

Sampling schedule:
- Control: at 0 and 48 hours
- Test concentration: at 0 and 48 hours

Storage
Test Item
- Recommended storage: Room temperature
- Storage at test facility: Room temperature, protected from moisture and light

Vehicle:

no

Details on test solutions:

Because the substance is insoluble in water Water Accommodated Fractions (WAF) were used to test effects at limit effective loadings and no specific analysis was performed. The content of the test item during the exposure period was verified by DOC determination.

Test Item
Stock solution: The stock solution (100 mg/L test item were weighed out) was prepared with dilution water in brown glass flask 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). Undissolved particles were removed by membran filtration (0.45 μm, RC, SCHLEICHER & SCHUELL).

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

Test organisms (species):

Daphnia magna

Details on test organisms:

- Test organism: Daphnia magna STRAUS (Clone 5)
- Origin: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu)
- Breeder: DR.U.NOACK-LABORATORIEN, 31157 Sarstedt, Germany
- Culture: 2-3 L glass vessels with approximately 1.8 L culture medium, at 20 ± 2 °C, in an incubator, 16 h illumination, illumination (max. 20µE·m-2 xs-1)
- Culture medium: Elendt M4, according to Elendt (1990), modified to a total hardness of 160 to 180 mg CaC03/L
- Feeding: At least 5 times per week ad libitum with a mix of Desmodesmus subspicatus and Chlorella vulgaris (cell density of > 10E-6 cells/mL)
- Number of test organisms per concentration and control: 20 animals, divided into 4 parallel samples, each with 5 animals
- Age of test organisms: 2 - 24 hold

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

14.3 °dH (256 mg/L CaCO3)

Test temperature:

20.3 °C

pH:

7.76 (mean value of 4 replicates after 48 h)

Dissolved oxygen:

8.67 (mean value of 4 replicates after 48 h)

Salinity:

n.a.

Nominal and measured concentrations:

100 mg/L of the test item corresponds to 1.2 mg/L DOC after 48 hours

Details on test conditions:

PRE-TREATMENT OF TEST ITEM AND PREPARATION OF TEST CONCENTRATIONS:
- The stock solution (100 mg/L test item were weighed out) was prepared with dilution water in brown glass flask one day prior to application and was shaken at room temperature with 20 rpm for 24 h (rotating shaker 3040, GFL)
- Undissolved particles were removed by membran filtration {0.45 µm, RC, SCHLEICHER & SCHUELL)
- The pH was measured to be 7.9

EXPOSURE CONDITIONS:
- Test vessels: glass beakers, holding 5 neonates in 20 ml of test medium
- Test concentration: 1 (100 mg/L), plus 1 control
- Number of neonates: 5 per vessel
- Acclimation: at least 2 h in dilution water
- Replicates: 4 per concentration/control
- Feeding during exposure: None
- Method of initiation: neonates were placed in prepared media
- Illumination: diffuse light, illumination range max. 20 µE·mE-2*sE-1
- Photoperiod: 16 h light:8 h dark
- Temperature of incubation unit: 18 to 22.0 +/- 1 °C
- Aeration: none
- Medium renewal: none
- Monitoring data: pH, dissolved oxygen and temperature, measured after 0 and 48 hours
- Criteria of effects: The criterion of adverse effects used in th is study was the item-induced alteration of the normal mobility behaviour and the loss of motory actions of the neonates, observed at 24 and 48 hours

Reference substance (positive control):

yes

Remarks:

Potassium dichromate p.a. (SIGMA) (Batch No. 065K1349; purity 99.9 %); Test concentrations: 1.66 - 1.99 - 2.39 - 2.87 mg/L

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The tested saturated solution* was visually clear throughout exposure. The percentage immobility was determined in the tested saturated solution* and the control after 24 h and 48 h. There is no biologically significant effect neither in the tested saturated solution nor in the control group.
*The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, acc. to OECD Series, No. 23 (2000)6

Results with reference substance (positive control):

The 24 h-EC50-value of 2.09 mg/L with 95 % confidence interval (1.98-2.21 mg/L) was calculated by analyses for reference item sigmoidal dose-response regression. Calculation of the confidence intervals for EC50 were carried out using standard procedures according to CLOPPER and PEARSON (1934 ). The range of validity was 1.0 mg/L < EC50 (24 h) < 2.5 mg/L.

Reported statistics and error estimates:

EC-values and statistical analyses for reference item
EC50-values was only calculated for the reference item by sigmoidal dose-response regression. Calculation of the confidence intervals for EC50 were carried out using standard procedures according to CLOPPER and PEARSON (1934 ).

Software: All data were computer generated and rounded for presentation from the full derived data. Consequently, if calculated manually based on the given data minor variations may occur from these figures. Calculations were carried out using software:
- GraphPad Prism4 (2005), GRAPHPAD SOFTWARE, INC.

DOC-Analysis

Analysis of New (0h) and Old Media (48h)

	DOC [mg/L]
	0 h			48 h			Overall
	Parallel determination		Mean	Parallel determination		Mean	Mean
Saturated solution	1.2	1.2	1.2	1.2	1.1	1.1	1.2
Control	1.3	1.3	1.3	1.3	1.3	1.3	1.3

Validity criteria fulfilled:

yes

Remarks:

(The immobilisation and other abnormalities in the controls did not exceed 10 % by the end of the test. The dissolved oxygen concentration remained above 3 mg/L throughout the exposure period)

Conclusions:

At the saturated solution of Solvent Violet 13 (1.2 mg/L DOC) no biologically significant effect was determined to Daphnia magna under static conditions over a period of 48 hours.

Executive summary:

ln the acute immobilization test with Daphnia magna (STRAUS) the effects of the saturated solution (1.2 mg/L DOC) of Solvent Violet 13, was determined according to OECD 202 (2004) and Directive 92/69/EC Method C (1992). The Iimit test was conducted under static conditions over a period of 48 hours. 20 test organisms were exposed to the saturated solution and the control. A reference test with potassium dichromate was carried out once per month to check the test system and test conditions. The EC50-value of the reference item of 2.09 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS. Water quality parameters pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable Iimits. The validity criteria of the test guideline were fulfilled. At the saturated solution of Solvent Violet 13 no biologically significant effect was determined. This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute Daphnia study.

Description of key information

ln the acute immobilization test with Daphnia magna (STRAUS) the effects of the saturated solution (1.2 mg/L DOC) of Solvent Violet 13, was determined according to OECD 202 (2004) and Directive 92/69/EC Method C (1992). The Iimit test was conducted under static conditions over a period of 48 hours. 20 test organisms were exposed to the saturated solution and the control. A reference test with potassium dichromate was carried out once per month to check the test system and test conditions. The EC50-value of the reference item of 2.09 mg/L after 24 h was within the prescribed concentration range of 1.0 to 2.5 mg/L according to AQS. Water quality parameters pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable Iimits. The validity criteria of the test guideline were fulfilled. At the saturated solution of Solvent Violet 13 no biologically significant effect was determined. This toxicity study is classified as acceptable and satisfies the guideline requirements for the acute Daphnia study.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

"Should read: > 100 mg/L (1.2 mg/L DOC)"

Solvent Violet 13 has been tested up to the limit of solubility in the test water. According to Directive 93/21/EEC ( adaptation to Directive 67/548/EEC ) the ecotoxicological classification is not toxic to Daphnia magna after 48 hours.