1H-Benz[e]indolium, 2-[2-[2-chloro-3-[2-[1,3-dihydro-1,1-dimethyl-3-(4-sulfobutyl)-2H-benz[e]indol-2-ylidene]ethylidene]-1-cyclohexen-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfobutyl)-, inner salt, ammonium salt (1:1)

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)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Test guideline 3

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14,2000.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

EC Number:

700-626-0

Cas Number:

1073607-50-7

Molecular formula:

C46 H50 Cl N2 O6 S2 . H4 N

IUPAC Name:

2-[(1E)-2-[(3Z)-2-chloro-3-{2-[(2E)-1,1-dimethyl-3-(4-sulfobutyl)-1H,2H,3H-benzo[e]indol-2-ylidene]ethylidene}cyclohex-1-en-1-yl]ethenyl]-1,1-dimethyl-3-(4-sulfonatobutyl)-1H-benzo[e]indol-3-ium amine

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Aanlysis performed at the start and after 48h of exposure.

Test solutions

Vehicle:

no

Details on test solutions:

Preparation of test solutions started with a loading rate of 100mg/l applying a 15-minute period of ultrasonic treatment in combination with stirring. Thereafter, the aqueous dispersion was magnetically stirred for three days to ensure reaching maximum dissolution in test medium. The resulting green dispersion contained precipitate and floating small particles at the surface and through the dispersion. The dispersion was consequently filtered througha 0.45µm membrane filter to remove the undissolved material. This resulted in a very slightly green filtrate that was used as highest test concentration. The lower test concentrations were prepared by subsequent dilutions of the filtrat in test medium. The dilutions were all clear and colourless.

Test organisms

Test organisms (species):

Daphnia magna

Study design

Test type:

static

Total exposure duration:

48 h

Test conditions

Hardness:

180mg/l expressed as CaCO3

Test temperature:

18-22°C

pH:

7.5-7.9

Dissolved oxygen:

8.7-8.8mg/l

Salinity:

The test medium is an adjusted ISO medium made by dissolving several macro salts, trace elements, macro nutrients and vitamins to tap water purified by reverse osmosis (medium called M7, as presribed by Dr. Elendt-Schneider: Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33)

Nominal and measured concentrations:

Analysis of duplicate samples taken from the filtered solution at the start of the test showed a measured concentration of 0.2mg/l. Measured concentrations in the dilutions were all below the limit of quantification. During the exposure period the measured concentration in the filtered solution decreased below the limit of quantification, i.e. below 0.1mg/l. This was expected to be due to the very low solubility in test medium, i.e. the start concentrations might stil be above the limit of solubility.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.32 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: immobility (including mortality)

Remarks on result:

other: The initial measured exposure concentration corresponds to a loading concentration of >100mg/l.

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.32 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: immobility (including mortality)

Remarks on result:

other: The initial measured exposure concentration corresponds to a loading concentration of >100mg/l.

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.32 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: immobility (including mortalilty)

Remarks on result:

other: The initial measured exposure concentration corresponds to a loading concentration of 100mg/l.

Any other information on results incl. tables

Acceptability of the test:

1.. In the control, no daphnids became immobilised.

2. The oxygen concentration at the end of the test was >+ 3mg/l in control and test vessels.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48h-EC50 exceeded an initial measured concentration of 0.32mg/l being the maximum soluble concentration of the test substance in test medium.
Due to the very low solubility in test medium concentrations toxic for Daphnia magna could not be reached.