Bis(N,N-dimethylpropane-1,3-diamine-N)[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]cobalt(1+) chloride

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:

2017-08-08 to 2017-10-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Bis(N,N-dimethylpropane-1,3-diamine-N)[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]cobalt(1+) chloride

EC Number:

284-943-5

EC Name:

Bis(N,N-dimethylpropane-1,3-diamine-N)[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]cobalt(1+) chloride

Cas Number:

84989-53-7

Molecular formula:

C42H44CoN12.Cl

IUPAC Name:

hydroxylamine

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: at test start and after 24 hours from fresh and aged solutions and after 48 hours from aged solutions.
- Fortified samples: stock solutions of the test item (9202 mg/L, 9580 mg/L and 1040 mg/L purity not considered) were prepared in methanol/water (1:1, v/v). The 9202 and 9580 mg/L solutions were used for fortification of 70 mg/L recovery samples. Dilutions (100 mg/L) were prepared in test medium and used for fortification of 1.8 mg/L recovery samples.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
- Stock solution: 50 mg of test item was mixed with test medium and homogenised by shaking for 15 minutes of sonication; Final volume is 1000mL
- Dilutions of stock solution: 3 dilutions were made from the stock solution with test medium, to reach a final volume of 455mL per dilution
- Controls: test water without addition of the test item

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: straus, clone 5
- Feeding during test: no feeding during test
- Feeding of culture (before test): three times a week with single cell green algae

Study design

Test type:

semi-static

Water media type:

freshwater

Remarks:

Elendt M4 medium

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

232 mg/L CaCO3

Test temperature:

19.0 – 20.6 °C, with a standard deviation of 0.4

pH:

7.85 – 8.30, with a standard deviation of 0.12

Dissolved oxygen:

9.3 mg/L

Nominal and measured concentrations:

Nominal concentrations: 50.0, 22.7, 10.3, 4.70 and 2.13 mg of test item/L and control
Measured concentrations (based on geometric mean): 12.6, 7.53, 3.14, 1.78, 0.900 and 0 mg of test item/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessel
- Type: covered with a glass plate (to reduce evaporation)
- Material, size, headspace, fill volume: 100 mL capacity filled with 50mL of test solution
- No. of organisms: 20 daphnids per test concentration (4 replicates of 5 animals each)
- No. of treatment groups: 6 (5 dose rates of the test item and a control)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4
- Culture medium different from test medium: same as Elendt M4

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours light/dark cycle

EFFECT PARAMETERS MEASURED: Immobilisation

RANGE-FINDING STUDY
- Test concentrations: 100, 50, 10 and 1mg/L, control
- Sampling points : at test start, at 24h and 48h
- Results used to determine the conditions for the definitive study:
- after 24h 5% of the daphnids were found to be immobile in the control, 10% at test item concentration of 50 mg/L and 15% at test item concentration of 100%
- after 48h 10% of the daphnids were found immobile in the control, 20% at test item concentration of 10 mg/L and 75 % at test item concentration of 50 mg/L and 65 % immobilisation was observed at the highest test item concentration of 100 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, two concentrations of this substance (1.00 mg/L, 2.00 mg/L) were tested around the same time period as the study

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

6.31 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.78 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Observations in control group: no immobilisation observed during test period

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Test concencentrations: 1 and 2 mg/L (20 organisms per test concentration)
- Relevant effect levels: EC50 (24h) = 1.48 mg test item/L

Any other information on results incl. tables

Biological results:

Results of the test, 48h values:

	nominal test item concentration [mg/L]
	Control	2.13	4.70	10.3	22.7	50.0
	immobilised daphnids after 48 h
Group 1	0	0	0	0	3	4
Group 2	0	0	0	2	3	3
Group 3	0	0	0	1	3	4
Group 4	0	0	0	1	4	5
Σ	0	0	0	4	13	16
%	0	0	0	20	65	80

Daphnids in all test concentrations were coloured blue and were clotted together. At the nominal test concentrations 10.3, 22.7 and 50 mg/L blue precipitation was observed at the bottom of the test vessel. The mobility of daphnids was however not influenced by the clotting nor by the formation of flakes or precipitates.

Analytical results:

Test item			Sampling	Test item		Test item concentration
nominal [mg/L]			[h]	[mg/L]	% of nominal	measured[mg/L]
Control			0 fresh	n.d.	-	0
			24 aged	n.d.	-
			24 fresh	n.d.	-
			48 aged	n.d.	-
2.13			0 fresh	< LOQ	-	0.900
			24 aged	< LOQ	-
			24 fresh	< LOQ	-
			48 aged	< LOQ	-
4.70			0 fresh	3.12	66	1.78
			24 aged	1.90	40
			24 fresh	1.90	40
			48 aged	< LOQ	-
10.3			0 fresh	6.05	59	3.14
			24 aged	4.86	47
			24 fresh	3.66	36
			48 aged	< LOQ	-
22.7			0 fresh	10.2	45	7.53
			24 aged	8.80	39
			24 fresh	7.07	31
			48 aged	5.07	22
50.0			0 fresh	23.3	47	12.6
			24 aged	9.80	20
			24 fresh	12.1	24
			48 aged	9.13	18

Because the measured concentrations (geometric mean) were used to derive the effect levels, the difference between 0h fresh and 24h fresh is less critical and does not impact the assessment.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EC50 for Daphnia magna is of 6.31 mg/L.

Executive summary:

The short-term toxicity to Daphnia magna was determined according to OECD 202 guideline with no deviations. The study was conducted under semi-static conditions. The daphnids were exposed over a period of 48 hours to different nominal concentrations of 50.0, 22.7, 10.3, 4.70 and 2.13 mg of the test item/L and a control. The test concentrations were analytically verified. Since the test concentrations decreased over time, results were expressed as geometric mean of measured concentrations. The nominal concentrations correspond to the measured ones 12.6, 7.53, 3.14, 1.78, 0.900 mg of test item/L and 0, respectively (geometric mean).

After 24h exposure no immobilization was observed in the control and up to and including 22.7 mg/L (nominal) and 15% was immobilized at 50 mg/L. After 48h exposure, no immobilisation was observed below and including 4.7 mg/L (nominal). 20 % of the daphnids were immobile at 10.3 mg/L, 65% at 22.7 mg/L and 80% at 50 mg/L. Daphnids in all test concentrations were coloured blue and were clotted together. At the nominal test concentrations 10.3, 22.7 and 50 mg/L blue precipitation was observed at the bottom of the test vessel. The mobility of daphnids was however not influenced by the clotting nor by the formation of flakes or precipitates.

The EC50 value was determined to be 6.31 mg/L (geometric mean of measured concentrations).