Bis[N-(4-{[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-ethylethanaminium] [mono and bis(dodecanyl, branched)]-(sulfonatophenoxy) benzenesulfonate

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:

2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

tetrakis(4-{[4-(diethylamino)phenyl][4-(ethylamino)naphthalen-1-yl]methylidene}-N,N-diethylcyclohexa-2,5-dien-1-iminium) 2-(dodecan-5-yl)-4-(4-sulfonatophenoxy)benzene-1-sulfonate 2-(dodecan-5-yl)-4-[3-(dodecan-5-yl)-4-sulfonatophenoxy]benzene-1-sulfonate

EC Number:

700-579-6

Molecular formula:

C90H114O7S2N6 (MW = 1454) and C102H138O7S2N6 (MW = 1622)

IUPAC Name:

tetrakis(4-{[4-(diethylamino)phenyl][4-(ethylamino)naphthalen-1-yl]methylidene}-N,N-diethylcyclohexa-2,5-dien-1-iminium) 2-(dodecan-5-yl)-4-(4-sulfonatophenoxy)benzene-1-sulfonate 2-(dodecan-5-yl)-4-[3-(dodecan-5-yl)-4-sulfonatophenoxy]benzene-1-sulfonate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

- Batch 310551

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 14 ml of test item diluted in 1 L liters of ultra-pure water (stock solution).
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): not mentionned

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia Magna

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

Room temperature

pH:

7.7 à 7.9 from T0 to T48h

Dissolved oxygen:

8.2 mg/L to 8.3mg/L at T0
7.2mg/L to 8.0 mg/L at T48H

Nominal and measured concentrations:

Nominal concentration:
0.18 - 0.11 - 0.063 - 0.036 - 0.021 - 0.011 - 0.007 - 0.0042 - 0.0025 - 0.0014 mg/L

Details on test conditions:

TEST SYSTEM
- Type of flow-through : static
- No. of organisms per vessel: 20
- No. of vessels per concentration : 4
- No. of vessels per control (replicates): 4

TEST CONCENTRATIONS
- Range finding study: 12.6 - 4.9 - 1.4 - 0.5 - 0.14 - 0.05 - 0.014 - 0.0049 mg/L
- Test concentrations: 0.18 - 0.11 - 0.063 - 0.036 - 0.021 - 0.011 - 0.007 - 0.0042 - 0.0025 - 0.0014 mg/L
- Results used to determine the conditions for the definitive study:
12.6 mg/L: 100% immobilisation
4.9 mg/L = 100% immobilisation
1.4 mg/L = 100 % immobilisation
0.5 mg/L = 100 % immobilisation
0.14 mg/L = 100 % immobilisation
0.05 mg/L = 90% immobilisation
0.014 mg/L = 20% immobilisation
0.0049 mg/L = 0% immobilisation

Reference substance (positive control):

yes

Remarks:

K2Cr2O7

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

ca. 0.012 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mobility

Remarks on result:

other: 95% CI : 0.0095-0.015

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 0.006 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mobility

Remarks on result:

other: 95% CI: 0.0047-0.0069

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.003 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mobility

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50 (24h) = 1.0 mg/L

Reported statistics and error estimates:

EC 50 was calculated by using Log-Probit statistical model (software Toxcalc)
NOEC was calculated by using the Bonferroni t-test (software Toxcalc).

Any other information on results incl. tables

ASSAY:

 

T = 24 hours
Concen- tration (mg/L)	0.18	0.11	0.063	0.036	0.021	0.011	0.007	0.0042	0.0025	0.0014	Control
I	0	0	0	0	2	3	5	3	5	5	5
II	0	0	0	1	1	3	2	4	5	5	5
III	0	0	1	1	2	4	3	5	5	5	5
IV	0	0	0	0	1	2	3	4	5	5	5
Total moving	0	0	1	2	6	12	13	16	20	20	20
Immobilization rate %	100%	100%	95%	90%	70%	40%	35%	20%	0%	0%	0%

 

T = 48 hours
Concen- tration (mg/L)	0.18	0.11	0.063	0.036	0.021	0.011	0.007	0.0042	0.0025	0.0014	Control
I	0	0	0	0	0	1	3	2	4	5	5
II	0	0	0	0	1	1	1	4	5	5	5
III	0	0	0	1	0	0	2	4	4	5	5
IV	0	0	0	0	0	0	1	4	5	5	5
Total moving	0	0	0	0	1	2	7	14	18	20	20
Immobilization rate %	100%	100%	100%	100%	95%	90%	65%	20%	0%	0%	0%

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the experimental conditions:
24-h EC50=0.012 mg/L (IC 95% : 0.0095; 0.015)
48-h EC50=0.0057 mg/L. (IC 95% : 0.0047; 0.0069)
48-h NOEC= 0.0025 mg/mL

Executive summary:

The short-term toxicity (48 hours) on invertebrate (Daphnia magna) of the test item Sepisol Fast Blue 85219 was assessed by determining the nominal effective concentration (EC 50%) at which 50 % of the test animals are immobilized. The method followed was the standard NF EN ISO 6341 under static condition.

 

EC50 (48h) values and associated confidence limits were calculated using a Log-Probit statistical model (software Toxcalc) and NOEC was worked out by using the Bonferroni t-test (software Toxcalc).

Test System: A stock solution (14 mg/mL) of the test substance was prepared in ultra-pur water.

Immobilisations and viable abnormalities were recorded at 24 and 48 hours as well as pH and dissolved oxygen.

Validity criteria were fulfilled and study was considered as valid..

A preliminary assay was performed at the following concentration: 12.6 - 4.9 - 1.4 - 0.5 - 0.014 - 0.0049 mg/L

Test Levels: Test concentrations included control, 0.18 - 0.11 - 0.063 - 0.036 - 0.021 - 0.011 - 0.007 - 0.0042 - 0.0025 - 0.0014 mg/L. Twenty daphnia were used per test group. 3 replicates were performed.

 

Conclusion:

 24-h EC₅₀=0.012 mg/L (IC 95%: 0.0095-0.015)

 48-h EC₅₀=0.0057 mg/L (IC 95%: 0.0047-0.0069)

48-h NOEC= 0.0025 mg/mL