Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)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:

2008/06/10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The reliabitity is rated 2 because the study followed a standard guideline of reference (NF EN ISO 6341), which describes a procedure designed to evaluate this endpoint. The results were reviewed and assessed as valid. However the study was not conducted under GLP conditions.

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:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Constituent 2

Reference substance name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

EC Number:

700-610-3

Molecular formula:

UVCB substance - not applicable

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Details on test material:

- Name of test material (as cited in study report): Sepisol Fast Blue 2BR
- Substance type:organic salt
- Physical state and appearance: dark blue powder
- Lot/batch No.: 905536
- Stability under test conditions: stable
- Storage condition of test material: at ambient temperature

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Water solubility (under test conditions): 73 mg/l and corresponding pH = 3,8 (with pure water)

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

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

Test conditions

Test temperature:

Room temperature

pH:

7.5 to 7.9 from To to T48h

Dissolved oxygen:

8.1 to 8.4 mg/L at To
7.4 to 8.1 mg/L at 48h

Nominal and measured concentrations:

8.0 - 4.6 - 2.7 - 1.6 - 0.9 - 0.55 - 0.34 - 0.19 - 0.11 - 0.061 - 0.039 - 0.023 mg/L and control

Details on test conditions:

TEST SYSTEM
- Type of flow-through (e.g. peristaltic or proportional diluter): static
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST CONCENTRATIONS
- Spacing factor for test concentrations: square root of 3
- Range finding study: 65.7 - 25.6 - 7.3 - 2.6 - 0.73 - 0.26 - 0.073 mg/L
- Test concentrations: 8.0 - 4.6 - 2.7 - 1.6 - 0.9 - 0.55 - 0.34 - 0.19 - 0.11 - 0.061 - 0.039 - 0.023 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr207)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24

Dose descriptor:

EC50

Effect conc.:

1.1

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI : 0.88 mg/L - 1.32 mg/L

Effect concentrations 2

Duration:

48

Dose descriptor:

EC50

Effect conc.:

0.13

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI: 0.10 mg/L - 0.17 mg/L

Effect concentrations 3

Duration:

48

Dose descriptor:

NOEC

Effect conc.:

0.023

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Williams' test

Results with reference substance (positive control):

- Results with reference substance valid? Results with reference substance, K2Cr2O7, are valids
- EC50/LC50: 1.0 mg/L

Reported statistics and error estimates:

EC50 calculated using the Log-Probit statistical model (Toxcalc software)
NOEC calculated using the Willams' Test (Toxcalc software).

Any other information on results incl. tables

Main Studies :

T = 24 h
Concentrations mg/L	8.0	4.6	2.7	1.6	0.9	0.55	0.34	0.19	0.11	0.061	0.039	0.023	Control
I	0	1	1	1	2	4	4	5	5	5	5	5	5
II	0	0	0	1	3	5	5	5	5	5	5	5	5
III	0	0	0	1	3	5	5	5	5	5	5	5	5
IV	0	0	1	2	2	4	5	5	5	5	5	5	5
Total moving	0	1	2	5	10	18	19	20	20	20	20	20	20
Immobilisation rate (%)	100%	95%	90%	75%	50%	10%	5%	0%	0%	0%	0%	0%	0%

Analytic serial number: 2469

Dissolved O₂at 8 mg/L: 8.0 mg/L

 

T = 48 h
Concentrations mg/L	8.0	4.6	2.7	1.6	0.9	0.55	0.34	0.19	0.11	0.061	0.039	0.023	Control
I	0	0	0	0	0	1	1	3	3	4	5	5	5
II	0	0	0	0	1	0	0	0	3	4	3	5	5
III	0	0	0	0	0	1	1	1	3	3	5	5	5
IV	0	0	0	0	0	0	1	2	4	4	3	5	5
Total moving	0	0	0	0	1	2	3	6	13	15	16	20	20
Immobilisation rate (%)	100%	100%	100%	100%	95%	90%	85%	70%	35%	25%	20%	0%	0%

Analytic serial number: 2469

Dissolved O₂at 1.6 mg/L: 7.4 mg/L

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the experimental conditions:
* 24h - EC50 = 1.1 mg/L (CI 95% : 0.88 - 1.32 mg/L)
* 48h - EC50 = 0.13 mg/L (CI 95% : 0.10 - 0.17 mg/L)
* 48h - NOEC = 0.023 mg/L

Executive summary:

The short-term toxicity (48h) on invertebrate (Daphnia magna) of the test item Sepisol Fast Blue 2BR was assessed by determining the nominal effective concentration (EC50) at which 50% of the Daphnia are immobilized. The method followed the standard NF EN ISO 6341 under static condition.

48h-EC50 values and associated confidence limits were calculated using a Log-Probit statistical model (software Toxcalc). The NOEC value was determined by using the Williams' test (software toxcalc).

The test substance was prepared in distilled water to obtain a stock solution of 73 mg/L. Immobilisation and viable abnormalities were recorded at 24h and 48h as well as pH and dissolved oxygen.

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

A preliminary assay was performed at the following concentrations: 65.7 - 25.6 - 7.3 - 2.6 - 0.73 - 0.26 - 0.073 mg/L

The main assay was performed at the following test concentrations: 8.0 - 4.6 - 2.7 - 1.6 - 0.9 - 0.55 - 0.34 - 0.19 - 0.11 - 0.061 - 0.039 - 0.023 mg/L + a control, in quadruplicate, with 20 daphnia per test and control groups.

Conclusion:

* 24h - EC50 = 1.1 mg/L (CI 95% : 0.88 - 1.32 mg/L)

* 48h - EC50 = 0.13 mg/L (CI 95% : 0.10 - 0.17 mg/L)

* 48h - NOEC = 0.023 mg/L