Dodeca (lithium, sodium)-2-({4-[4-(4-{bis[alkyl-(sulfonatoalkoxy)-4-({-sulfonato-[(substituted-phenyl)diazenyl]phenyl} diazenyl)anilino]-triazin-yl}-6-[alkyl-(sulfonatoalkoxy)-4-(sulfonato--[(substituted-phenyl)diazenyl]phenyl)diazenyl)anilino]piperazin-yl)-triazin-ylamino]-alkyl-5-(sulfonatoalkoxy)phenyl}diazenyl)-5-[(sulfonatophenyl)diazenyl]benzensulfonate

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:

22-24 December 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted following the GLP principles and according to the OECD and Japanese guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

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:

other: "Alga growth inhibition test, Daphnia sp. acute immobilisation test, and fish acute test" Yakushokuhatsu No. 1121002, Heisei 15.11.13 Seikyoku No. 2, Kanpokihatsu No. 031121002, November 21, 2003; the latest revision November 20, 2006.

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): E-BW102
- Substance type: reddish-brown powder
- Physical state: solid
- Lot/batch No.: MB-1
- Storage condition of test material: room temperature, shaded, air-tight

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 10,18, 32, 56, 100
- Sampling method: from the middle layer of one vessel at teh time of preparation and at the end of exposure
- Sample storage conditions before analysis: not reported

Test solutions

Vehicle:

no

Details on test solutions:

The test solutions were prepared just before exposure. First, 200 mg test substance was dissolved in 2L dilution water resulting in a concentration of 100 mg/L. this was diluted further with dilution water resulting in concentrations of 56, 32, 18 and 10 mg/L. All these solutions were made in a final volume of 500 ml.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:Daphnia
- Strain: Daphnia magna
- Source: National Institute for Environmental Studies, Japan
- Age at study initiation (mean and range, SD): < 24 h old juvenile females
- Feeding during test: none


ACCLIMATION
- Acclimation period: 48 h (for the parental Daphnida that produced the juveniles)
- Acclimation conditions (same as test or not): yes
- Type and amount of food: unicellular green algae, Chlorella vulgaris
- Feeding frequency: 6 mg C/2 L/ day for 25 daphnids
- Health during acclimation (any mortality observed): < 20 % mortality in parental daphnids 2 weeks prior the start of the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

54 mg/L CaCO3

Test temperature:

19.8 - 20.3

pH:

7.7 - 7.9

Dissolved oxygen:

8.4 - 8.8 mg/L

Nominal and measured concentrations:

Nominal: 10, 18, 32, 56, 100 mg/L
Measured: 8.64, 17.0, 29.5, 52.8, 96.4 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Type: closed
- Material, size, headspace, fill volume: glass, 100 ml
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated Yokohama city tap water
- Total organic carbon: < 0.3 mg/L
- Particulate matter: < 1 mg/L
- Metals: Al 0.05 mg/L
- Pesticides: < 0.001 mg/L
- Chlorine: < 0.1 mg/L
- Alkalinity: 37 mg CaCO3/L pH 4.8
- Ca/mg ratio: 14/3.3
- Conductivity: 13 mS/m
- Culture medium different from test medium: no
- Intervals of water quality measurement: every 6 months

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/ 8 h darkness
- Light intensity: 800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : median effect concentration for immobility

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: yes
- Test concentrations: 1.0, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: 10% immobility was recorded after 48 h exposure to 100 mg/L in the range finding test.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

the measured concentrations were within 20% of nominal

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: no
- Observations on body length and weight: not reported
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no precipitation, floating solids, suspended solids
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: yes
- EC50: 0.81 mg/L (95 C.I. 0.75-0.87 mg/L)
- Other: the sensitivity tests were performed at every 6 months. the above results were the most recent in regard to the date of the experiment start.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this experiment, E-BW 102 did not induce acute immobilisation in daphnids up to 100 mg/L (EC50 > 100 mg/L).