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Ecotoxicological information

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:
7.7.2010 - 23.8.2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was carried out according to recommended method in accordance with internationally valid GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
yes
Remarks:
See section Overall remarks
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Copper phtalocyanine monosulfonated and disulfonated
EC Number:
700-870-8
Molecular formula:
C32H15N8Cu(HSO3)n where n=1.3-1.6
IUPAC Name:
Copper phtalocyanine monosulfonated and disulfonated
Constituent 2
Reference substance name:
Copper phtalocyanine mono- and disulphonated
IUPAC Name:
Copper phtalocyanine mono- and disulphonated
Constituent 3
Reference substance name:
Hysperse 12
IUPAC Name:
Hysperse 12
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Copper phtalocynine mono- and disulphonated
- Molecular formula: C32H15N8Cu(SO3H)1.4x2.5 H20
- Molecular weight: 733.3
- Substance type: technical product
- Physical state: powder
- Lot/batch No.: 01
- Expiration date of the lot/batch:
- Stability under test conditions: stable
- Storage condition of test material: The substance was stored in closed vessel in dry room at the temperature bellow 25ºC in dark.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Limit test: The analytical determination of the test substance concentration were performed at the beginning and at the end of the test. The samples for analysis were taken from the highest (100 mg.L-1) test concentration. The sample for analysis (0 hours) was prepared at the beginning of the test and immediately delivered in transport box to analytical laboratory.
Preliminary test: The analytical determination of the test substance concentrations was performed at the beginning and at the end of the test. The samples for analysis were taken from the highest (100 mg.L-1) and lowest (1 mg.L-1) test concentrations. The samples for analysis (0 hours) were prepared at the beginning of the test and immediately delivered in transport box to analytical laboratory. The samples were analyzed on the day of delivery. The samples for analysis after 48 hours were delivered to analytical laboratory immediately after the end of testing. The samples were analyzed on the day of delivery. All samples were stored at laboratory temperature.
Full test: The analytical determination of the test substance concentrations was performed at the beginning and at the end of the test. The samples for analysis were taken from the highest (100 mg.L-1) and lowest (10 mg.L-1) test concentrations The samples for analysis (0 hours) were prepared at the beginning of the test by Analytical Group I and immediately delivered in transport box to Ecotoxicity laboratory. The samples were analyzed on the day of preparation.
The samples for analysis after 48 hours were delivered to analytical laboratory immediately after the end of testing. The samples were analyzed on the day of delivery. All samples were stored at laboratory temperature.

Test solutions

Vehicle:
no
Details on test solutions:
The stock solution of the test substance was prepared in the dilution water. 100 mg of the test substance was weighed into 1000 mL of the dilution water for the limit test.
The concentrations of solutions used in the preliminary test and full test were obtained by dilution of the stock solution with dilution water.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnia
- Strain: Daphnia magna Straus
- Source: own laboratory breeding
- Age at study initiation: < 24 h
- Method of breeding:
The grow-up gravid females are separated into dilution water. Young daphnia born in 24 hours are segregated to the handling container from which they entered the experiment. At handling a care should be taken of not to penetrate the air under animal’s shell.
- Feeding during test: no
- Aeration during test: no

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
none

Test conditions

Hardness:
Content of (Ca+Mg) in dilution water: 2.49 mmol/L limit test)
Content of (Ca+Mg) in dilution water: 2.52 mmol/L (preliminary test)
Content of (Ca+Mg) in dilution water: 2.49 mmol/L (full test)

Test temperature:
20.3 = 22.0 °C (limit test)
20.0 - 20.1ºC (preliminary test)
20.2 - 20.0ºC (full test)

pH:
pH = 7.3 - 7.8 (limit test)
pH = 7.4 - 7.9 (preliminary test)
pH = 7.8 -8.7 (full test)

Dissolved oxygen:
Concentration of dissolved oxygen 8.1 - 8.4 mg/L (limit test)
Concentration of dissolved oxygen 7.5 - 8.0 mg/L (preliminary test)
Concentration of dissolved oxygen 7.8 - 8.7 mg/L (full test)

Nominal and measured concentrations:
Loading rate limit test: 100mg/L (nominal), 92.5 and 89.5 (measured)
Change of concentration in interval 0 - 48 h at 100.0 mg/l (nominal): -3.2 %
Loading rate preliminary test : 100, 50, 10, 5, 1 mg/L (nominal)
Change of concentration in interval 0 - 48 h at 1.0 mg/l (nominal): 0 %
at 100 mg/L (nominal): -3.8 %
Loading rate full test: 100, 80, 53, 35, 23, 15, 10 mg/L (nominal)
Change of concentration in interval 0 - 48 h at 10 mg/l (nominal): -13.7 %
at 100 mg/L (nominal): -8.6 %


Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type: open beaker
- Material: glas
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Standard dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D into 1000 mL of deionised water.
A. 117.6 g CaCl2.2H20 in 1 L of deionised water
B. 49.3 g MgSO4.7H2O in 1 L of deionised water
C. 25.9 g NaHCO3 in 1 L of deionised water
D. 2.3 g KCl in 1 L of deionised water
The dilution water should be of pH 7.8 ± 0.2.
The sum of Ca + Mg ions should be approx. 2.5 mmol/L.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Lighting: natural variation of daily light and dark
- Light intensity: daily

EFFECT PARAMETERS MEASURED: immobilization of daphnia



Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
24.7 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% confidence limit: 14.2 – 35.1 mg/L
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
21.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% confidence limit:12.0 – 30.8 mg/L
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
24 h
Dose descriptor:
EC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC100
Effect conc.:
80 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: no
- Observations on body length and weight: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses:no

Results with reference substance (positive control):
- Results with reference substance are valid
The results of the verification test with K2Cr2O7, carried out in period from03.10.2012. to 05.10.2012 were following:

24h – EC50 = 1.5 mg/L (95 % confidence limit: 1.29 – 1.70 mg/L)
48h – EC50 = 0.83 mg/L (95 % confidence limit: 0.71 – 0.96 mg/L)

The result of the last interlaboratory test arranged by CSlab spol. s r. o. in 2013for potassium dichromate was:
48-hour EC50 = 0.58 – 1.34 mg/L
The immobilisation of test organism by refernece substance obtained from last reference test meets the calculated range from interkaboratory test.
Reported statistics and error estimates:
The value of EC50 was calculated by means of software “Toxicita” (product of Water Research Institute, Ostrava, Czech Republic). The NOELR and EL100 values are identified by direct observation.
The loading rates of the test substance were used for all evaluations and results.

Any other information on results incl. tables

Limit test

Table 1.Immobilisation of daphnia – limit test

Nominal concentration

 mg·L-1

 100

 C

Number of mobile animals

 24 hours/1stseries

 3

 10

Number of mobile animals

 24 hours/2ndseries

 3

 9

Number of mobile animals

24 hours/total

 6

 19

Immobilisation %

24 hours

70

 5

Number of mobile animals

 48 hours/1stseries

 1

 10

Number of mobile animals

 48 hours/2ndseries

 0

 8

Number of mobile animals

 48 hours/total

 1

 18

Immobilisation %

 48 hours

 95

 10

 

Preliminary test

Table 5.Immobilisation of daphnia – preliminary test

Nominal concentration

mg·L-1

 100

 50

 10

 5

 1

 C

Number of mobile animals

 24 hours/1stseries

 0

 5

 9

 10

 10

 10

Number of mobile animals

 24 hours/2ndseries

 0

 7

 10

 10

 5

 10

Number of mobile animals

24 hours/total

 0

 12

 19

 20

 15

 20

Immobilisation %

 24 hours

 100

 40

 5

 0

 25

 0

Number of mobile animals

 48 hours/1stseries

 0

 3

 9

 10

 10

 10

Number of mobile animals

 48 hours/2ndseries

 0

 3

 8

 10

 5

 10

Number of mobile animals

 48 hours/total

 0

 6

 17

 20

 15

 20

Immobilisation %

 48 hours

 100

 70

 15

 0

 25

 0

 

Full test

Table 9.Immobilisation of daphnia – full test

 

Nominal concentration

 mg·L-1

 100

 80

 53

 35

 23

 15

10

 C

Number of mobile animals

 24 hours/1st series

 1

 0

 1

 2

 5

 10

 9

 10

Number of mobile animals

 24 hours/2nd series

 0

 1

 1

 7

 3

 8

 10

 10

Number of mobile animals

 24 hours/total

 1

 1

 2

 9

 8

 18

19

 20

Immobilisation %

 24 hours

 95

 95

 90

 55

 60

 10

 5

 0

Number of mobile animals

48 hours/1st series

 0

 0

 1

 2

 3

 9

 9

 10

Number of mobile animals

 48 hours/2nd series

 0

 0

 1

 4

 3

 8

 10

 9

Number of mobile animals

 48 hours/total

 0

 0

 2

 6

 6

 17

 19

 19

Immobilisation %

  48 hours

 100

 100

 90

 70

 70

 15

 5

 5

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
see Overall remarks
Executive summary:

The substance -Hysperse 12- was tested in acute immobilisation test on Daphnia magna.

The test was performed according to method C.2 Daphnia sp. Acute Immobilisation Test, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

The limit test, preliminary test and full test were performed as static.

 

The study was performed with the analytical determination of the test substance concentrations. Concentration of the test substance was satisfactorily maintained within ± 20 per cent of the nominal or measured initial concentration throughout the test. So the nominal concentrations were used for all evaluations and results.

 

Test results:

24 hour-EC50 = 24.7 mg/L   (95% confidence limit: 14.2 - 35.1 mg/L)   (nominal concentration)  

48 hour-EC50 = 21.4 mg/L   (95% confidence limit: 12.0 – 30.8 mg/L)  (nominal concentration)

   

24 hour-NOEC = 10.0 mg/L          (nominal concentration)

48 hour-NOEC = 10.0 mg/L          (nominal concentration)

 

24 hour-EC100 > 100 mg/L (nominal concentration)

48 hour-EL100 = 80 mg/L          (nominal concentration)