Cuprate(4-), [[3,3',3'',3'''-[(29H,31H-phthalocyanine-1,8,15,22-tetrayl-.kappa.N29,.kappa.N30,.kappa.N31,.kappa.N32)tetrakis(sulfonyl)]tetrakis[1-propanesulfonato]](6-)]-, sodium (1:4), (SP-4-1)-

Toxicity to aquatic plants other than algae

Administrative data

Endpoint:

toxicity to aquatic plants other than algae

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental phase 2013-11-29 to 2013-12-23

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Meets the criteria for classification as relaible without restriction according to Klimisch et al (1997).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 221 (Lemna sp. Growth Inhibition Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

tetrasodium 1,8,15,22-tetra-(3-sulfoxypropylsulfonyl)copper phthalocyanine

EC Number:

696-145-8

Cas Number:

944730-39-6

Molecular formula:

C44H36CuN8O20S8 .4Na

IUPAC Name:

tetrasodium 1,8,15,22-tetra-(3-sulfoxypropylsulfonyl)copper phthalocyanine

Test material form:

other: solid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 120 mg/l test material
- Sample storage conditions before analysis: Day 7 samples were analysed on the day of receipt while the day 0 samples were frozen prior to analysis

Test solutions

Vehicle:

no

Details on test solutions:

An amount of test item (240 mg) was dissolved in culture medium and the volume adjusted to 2 liters to give the required test concentration of 120 mg/L. The prepared concentration was inverted several times to ensure adequate mixing and homogeneity. The concentration and stability of the test item in the test preparations were verified by chemical analysis on Day 0 (fresh media) and Day 7 (old media).

Test organisms

Test organisms (species):

Lemna gibba

Details on test organisms:

The test was carried out using Lemna minor. A culture of Lemna minor was obtained from Canadian Phycological Culture Centre, University of Waterloo, Ontario, Canada. Cultures were maintained in the laboratory by the periodic replenishment of culture medium. The culture was maintained in the laboratory at a temperature of 24 ± 2 ºC under continuous illumination (intensity approximately 7000 lux) for at least 7 days prior to the start of the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

7 d

Test conditions

Test temperature:

24 +/- 2ºC

pH:

6.6-7.6

Nominal and measured concentrations:

Chemical analysis of the 120 mg/L test preparation on Day 0 (fresh media) and Day 7 (old media) showed measured test concentrations of 102% and 104% of nominal respectively and so the results are based on nominal test concentrations only.

Details on test conditions:

TEST SYSTEM
- Test vessel: Three conical flasks each containing 250 mL of solution were prepared for the control and six flasks for the treatment group.
- No. of colonies per vessel: 3
- No. of fronds per colony: 3


GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:


OTHER TEST CONDITIONS
The flasks were then incubated at 24 ± 2 ºC under constant illumination (intensity approximately 7000 lux) for 7 days.

EFFECT PARAMETERS MEASURED
The number of fronds present in each test and control culture was recorded on days 0, 2, 5 and 7 along with observations on frond size, appearance, root length and number of colonies present.

In addition the dry weight of the fronds in each control and treatment group was determined on day 7. At the start of the test six replicate samples of fronds identical to those used to inoculate the test vessels were taken and the dry weight determined. At the end of the test dry weight of colonies from each control and test vessel was determined by blotting the colonies dry and drying at 60 °C to constant weight.


RANGE-FINDING STUDY
- Test concentrations: 120 mg/l
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

3,5-Dichlorophenol

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

7 d

Dose descriptor:

EC20

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 4

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 5

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Effect concentrations 6

Duration:

7 d

Dose descriptor:

EC20

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Effect concentrations 7

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Effect concentrations 8

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Effect concentrations 9

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 10

Duration:

7 d

Dose descriptor:

EC20

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 11

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 12

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 13

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 14

Duration:

7 d

Dose descriptor:

EC20

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 15

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 16

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Any other information on results incl. tables

A positive control used 3,5-dichlorophenol as the reference item at concentrations of 0.625, 1.25, 2.5, 5.0 and 10 mg/L.

 

Exposure conditions and data evaluation for the positive control were similar to those in the definitive test.

 

Exposure of Lemna minor to the reference item gave the following results:

 

Response Variable	Measurement Variable	EC50(mg/L)	95% Confidence Limits	No Observed Effect Concentration (NOEC) (mg/L)	Lowest Observed Effect Concentration (LOEC) (mg/L)
Average Specific Growth Rate	Frond Number	3.7	3.3 – 4.0	1.25	2.5
	Dry Weight	2.8	2.6 – 3.0	1.25	2.5
Yield	Frond Number	3.0	2.7 – 3.3	1.25	2.5
	Dry Weight	2.0	1.8 – 2.2	1.25	2.5

 

The results from the positive control with3,5-dichlorophenol were within the normal ranges for this reference item.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The average specific growth rate EC50 for frond number and dry weight was >120 mg/l. On this basis the test substance is considered to be non-toxic to Lemna gibba in this study.

Executive summary:

Introduction

This study was undertaken to assess the effect of the test item on the growth of the freshwater plant Lemna minor. The method followed that described in the OECD Guideline No 221 'Lemna sp. Growth Inhibition Test (March 2006)'

Results & Conclusion

The average specific growth rate EC50 for frond number and dry weight was >120 mg/l. On this basis the test substance is considered to be non-toxic to Lemna gibba in this study.