1-Octadecanaminium,N,N-Dimethyl-N-Octadecyl-,(Sp-4-2)-[29h,31h-Phthalocyanine2-Sulfonato(3-)-N 29,N 30,N 31,N 32]Cuprate(1-)

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study; well documented study report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Physical state: Blue powder
- Analytical purity: 96.3%
- Composition of test material, percentage of components: NDA
- Storage condition of test material: At room temperature in the dark in the container in which it was received.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

-Concentrations: the concentration of the test material in the test solutions were impossible to be verified by chemical analysis due to its low solubility in organic solvents other than dimethylformamide (DMF).
-Sampling method: at the start of the test, a sample of the stock was taken and analyzed for the concentration of the test substance; at the end of the test each blank solution was sampled and analyzed in the same manner.
- Sample storage conditions before analysis: No data.

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Nominal exposure concentration preparations.
- Controls: Blank control and solvent control.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethylformamide (DMF)
- Concentration of vehicle in test medium (stock solution and final test solution): 0.1 ml DMF/L in culture medium.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The test solutions were clear by visual assessment

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Selenastrum capricornutum
- Strain: ATCC 22662
- Source (laboratory, culture collection): From laboratory cultures maintained under axenic conditions.
- Age of inoculum (at test initiation): A culture of the alga in the exponential growth phase was used as inoculum of the test.
- Method of cultivation: No data.
ACCLIMATION
- Acclimation period: No data.
- Culturing media and conditions (same as test or not): Yes.
- Any deformed or abnormal cells observed: No data.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Hardness:

No data

Test temperature:

The daily temperature measurements recorded in the incubator ranged from 23.3 to 23.9 °C. The hourly temperature measurements recorded automatically, ranged from 24.4 to 25.0 °C.

pH:

At the start of the test, the pH of all the test solutions was 7.4, and at the end of the test the range was 7.9-8.1.

Dissolved oxygen:

No data

Salinity:

No data

Nominal and measured concentrations:

Nominal concentrations: 0.010, 0.018, 0.032, 0.056, 0.1 mg/L, with a control and solvent control

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: The test vessels were borosilicate glass conical flasks of 250 ml nominal capacity closed with polyurethane foam bungs. Each algal test falsk contained 100 ml of test sulutions.
- Initial cells density: 1.03 x 104 cells /ml.
- Control end cells density: 1.03 x 104 cells /ml.
- No. of organisms per vessel: 1.03 x 106 cells in 100 ml media.
- No. of vessels per concentration (replicates): triplicate each concentration of the test substance.
- No. of vessels per control (replicates): triplicate cultures of the culture medium control
- No. of vessels per vehicle control (replicates): 6 replicate cultures of the solvent control
- Other: One blank vessel (without algal inoculum) for each control and treatment was incubated concurrently.
GROWTH MEDIUM
- Standard medium used: yes
OTHER TEST CONDITIONS
- Sterile test conditions: Yes.
- Light intensity and quality: measured once during the study, was 8420 lux (by cosine receptor). This was also measured in terms of quantum response and was 66.4 microeinsteins/m2/sec
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: electronic particle counter.

TEST CONCENTRATIONS
No range finding study was performed.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

exceeding solubility

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

exceeding solubility

Effect concentrations 3

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks:

exceeding solubility

Effect concentrations 4

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks:

exceeding solubility

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

No data

Any other information on results incl. tables

Table 1. Algal Cell Densities

Nominal Conc. of Test Material ( mg/L)	Algal cell density (x 10^4cell/ml) (Mean)
	Day 1	Day 2	Day 3
Control (N = 3)	6.07	38.2	211
Solvent Control (N = 6)	6.06	38.9	201
0.01	5.44	38.2	204
0.018	5.12	33.1	193
0.032	5.36	38.4	207
0.056	5.58	35.6	201
0.10	5.34	34.2	184

Table 2. Mean Areas Under The Growth Curve, And Percentages Of The Solvent Control Indicating Statistically Significant Differences

Nominal Conc. of Test Material ( mg/L)	Mean Area under Growth Curve (0-72 h)	% of Solvent Control
Control	147.1	103
Solvent Control	143.1	-
0.010	142.9	100
0.018	132.1	92
0.032	144.7	101
0.056	139.1	97
0.10	128.9	90

There were no statistically significant differences from the solvent control (P = 0.05).

Table 3. Man Growth Rates Over The Test Period, And Percentages Of The Solvent Control Indicating Statistically Significant Differences

Nominal Conc. of Test Material ( mg/L)	Mean Growth Rate (0-72 h)	% of Solvent Control
Control	1.774	101
Solvent Control	1.758	-
0.010	1.726	100
0.018	1.744	99
0.032	1.768	101
0.056	1.758	100
0.10	1.728	98

There were no statistically significant differences from the solvent control (P = 0.05).

Table 4. Test Solution pH Measurements

Nominal Conc. of Test Material ( mg/L)	Day 0	Day 3
		Replicate A	Replicate B
Control	7.4	8.0	8.1
Solvent Control	7.4	8.1	8.0
0.010	7.4	8.0	8.0
0.018	7.4	7.9	8.0
0.032	7.4	8.0	8.1
0.056	7.4	7.9	8.1
0.10	7.4	7.9	8.0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The EC values based on the 72 h nominal test concentration were greater than 0.1 mg/L, correspondingly the NOEC was greater than or equal to 0.1 mg/L based on areas under the growth curve, or on logarithmic growth rate over the test period.

Executive summary:

In a 72 h acute toxicity study, the cultures of the green algaSelenastratum capricornutumwere exposed to the test material at a nominal concentrations of 0.010, 0.018, 0.032, 0.056, 0.1 mg/L, with a control and solvent control, under static conditions in accordance with OECD 201.  The NOEC and EC₅₀values based on logarithmic growth rate over the test period were 0.10 and >0.10 mg/L (nominal), respectively.  The NOEC and EC₅₀values based on areas under the growth curve were 0.10 and >0.10 mg/L (nominal), respectively. There were no compound related phytotoxic effects.