Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, sulfonated

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study from supporting substance (structural analogue)

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)

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.

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 (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

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

none

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

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

Key result

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

Key result

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.

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/1^stseries	0	5	9	10	10	10
Number of mobile animals	24 hours/2^ndseries	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/1^stseries	0	3	9	10	10	10
Number of mobile animals	48 hours/2^ndseries	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

 

Validity criteria fulfilled:

yes

Remarks:

see Overall remarks

Conclusions:

Scute immobilisation test on Daphnia magna, performed according to method C.2 Daphnia sp. Acute Immobilisation Test, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008, resulted in the following:
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)    

Executive summary:

The substance was tested in acute immobilisation test on Daphnia magna, 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)    

Description of key information

The substance was tested in acute immobilisation test on Daphnia magna, 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)    

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

21.4 mg/L