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EC number: 275-060-6 | CAS number: 70955-64-5
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_ecotoxicological-information.png)
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:
- from 2018-06-20 to 2018-06-28, with the definitive exposure phase from 2018-06-20 to 2018-06-27
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
- Version / remarks:
- 2006
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Council Regulation (EC) No. 761/2009 Method C.26
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, [[2-[(4-chloro-6-methoxy-1,3,5-triazin-2-yl)amino]ethyl]amino]sulfonyl sulfo derivs., sodium salts
- EC Number:
- 275-060-6
- EC Name:
- Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, [[2-[(4-chloro-6-methoxy-1,3,5-triazin-2-yl)amino]ethyl]amino]sulfonyl sulfo derivs., sodium salts
- Cas Number:
- 70955-64-5
- Molecular formula:
- C36.2H18.9Cl0.7CuN11.5O10.2S3.4Na2.7
- IUPAC Name:
- Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, [[2-[(4-chloro-6-methoxy-1,3,5-triazin-2-yl)amino]ethyl]amino]sulfonyl sulfo derivs., sodium salts
- Test material form:
- solid: particulate/powder
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- Determination of the test item
All test item concentrations and the control were analytically verified via HPLC-DAD at the first interval. The control, the lowest and the highest test item concentration were analytically verified at the end of the exposure (7 days) and on every renewal day. Freshly prepared and old media were analyzed. The samples were analyzed with an HPLC-DAD method. The method was implemented under non-GLP but documented in the raw data and validated. The method validation was not part of this GLP study.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- Preparation of the Test item solution
A stock solution with a nominal test item concentration of 100 mg/L was freshly prepared with dilution water and agitated until the solution was visually clear. The test item solution was clear and blue colored.
Test concentrations
Based on the results of a preliminary range finding test (see section 18) 5 concentrations were tested with a dilution factor of √10: 1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L.
Control
Six replicates (without test item) were tested under the same test conditions as the test vessels.
Test organisms
- Test organisms (species):
- Lemna gibba
- Details on test organisms:
- Test organism
Duckweed, Lemna gibba G3, Lemnaceae, Arales, Arecidae, Monocotyledonae
Young, rapidly growing plants without visible lesions or discolouration (chlorosis) were used for the test.
Reason for the selection of the test organism
According to the guideline, Lemna gibba is a suitable species because it is a representative of temperate areas commonly used for toxicity tests.
Origin
EUROFINS-GAB GMBH, Eutinger Str. 24, 75223 Niefern-Öschelbronn, Germany
Cultivation at test facility
The species is cultured in the test facility. Density is kept low to prevent conglomerates of plants on the surface. At least once per week, plants are transferred to freshly prepared growth medium. Growth media and culturing vessels are autoclaved before use to enable the breeding of axenic cultures.
Breeding vessels
Crystallisation dishes of glass, vol. 900 mL, filled with ca. 500 mL growth medium, covered with glass tops
Medium
20X-AAP-medium (Algal Assay Procedure medium),
pH-value 7.5 ± 0.1
Composition of Dilution water
Component Concentration in stock solution [g/L] Concentration in prepared medium [mg/L]
NaNO3 26 510
MgCl2 x 6 H2O 12 240
CaCl2 x 2 H2O 4.4 90
MgSO4 x 7 H2O 15 290
K2HPO4 · 3 H2O 1.4 30
NaHCO3 15 300
H3BO3 0.19 3.7
MnCl2 x 4 H2O 0.42 8.3
FeCl3 x 6 H2O 0.16 3.2
Na2-EDTA · 2 H2O 0.30 6.0
ZnCl2 3.3 mg/L 66 µg/L
CoCl2 x 6 H2O 1.4 mg/L 29 µg/L
Na2MoO4 x 2 H2O 7.3 mg/L 145 µg/L
CuCl2 x 2 H2O 0.012 mg/L 0.24 µg/L
pH-value 7.5 ± 0.1
The pH of the test medium had to be 7.5 +/- 0.1 and was adjusted prior to testing with the addition of 1 N NaOH and HCl.
Temperature 24 ± 2 °C
Light regime
Continuous fluorescent light, 1100 – 4440 lux
Acclimatization of the test system
The test system (the test organism) was held for 7 days under test conditions to acclimatize. These acclimatized plants were used in the test.
Study design
- Test type:
- semi-static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 7 d
Test conditions
- Hardness:
- not measured
- Test temperature:
- Room temperature [°C]: min.: 22.2 max.: 23.1 mean value: 22.5
- pH:
- Nominal test item concentration
[mg/L] pH-value
0 h (Fresh medium) 2 d (Old medium) 2 d (Fresh medium) 5 d (Old medium) 5 d (Fresh medium) 7 d (Old medium)
100 7.55 8.39 7.56 8.58 7.85 8.57
31.6 7.55 8.39 7.55 8.59 7.85 8.60
10.0 7.54 8.38 7.54 8.59 7.85 8.62
3.16 7.55 8.37 7.53 8.58 7.83 8.59
1.00 7.58 8.36 7.53 8.57 7.83 8.58
Control 7.59 8.36 7.54 8.58 7.84 8.57 - Dissolved oxygen:
- not measured
- Salinity:
- not measured
- Conductivity:
- not measured
- Nominal and measured concentrations:
- Nominal: 1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L
- Details on test conditions:
- Test method
Semi-tatic procedure
Test duration
7 days
Replicates
3 replicates per concentration level, 6 for the control.
Test vessels/test volumes
Crystallisation dishes with a volume of 500 mL, covered with glass tops and filled with 200 mL test solution were used in the test. The test vessels were placed on a black non-reflective surface to avoid stray light.
Dilution water
20X-AAP-medium according to the guideline.
Application
Semi-static, with renewal of the test solutions on day 2 and 5 (i.e. exposure to freshly prepared test and control solutions on two occasions during the test). The plants were placed into the test vessels, which already contain the test media.At the start of the exposure, 3 uniform, healthy plants (colonies of 4 fronds each), were introduced into each test vessel containing the test media. The initial frond number per test vessel was 12. The initial numbers of colonies and fronds were the same in each test vessel.
Temperature (Target)
24 ± 2 °C
Light regime (Target)
Continuous, fluorescent light, 6500 to 10000 lux on the surface of the test medium (difference of light intensity at any measured incubation place < 15 % from the mean value)
Placement of the test vessels
A randomised placement of the test vessels was carried out.
Type and frequency of measurements
The numbers of plants and fronds were determined at the start and
the end of the exposure. The number of fronds was determined every 2 - 3 days from each replicate of the control and the test concentrations. Every frond that visibly projected beyond the edge of a parent frond was counted as a separate frond. Fronds that lost their pigmentation were not counted.
Observations of frond size, appearance, indication of necrosis, chlorosis or gibbosity, colony break-up or loss of buoyancy, of root length and appearance, as well as of change in colour and destruction of roots, were made on every determination day and at the end of the exposure.
After 7 days, the determination of dry weight was carried out from 3 replicates per test concentration and 6 control replicates. Colonies from each test vessel were collected, rinsed with deionised water and then dried at 60 °C to a constant weight. Any root fragments were included. The dry weight was expressed to an accuracy of
0.1 mg.
The dry weight of the starting biomass was determined based on a sample of fronds (same number of fronds as in the test vessels) taken from the same batch used to inoculate the test vessels.
Physico-chemical Parameters
The pH-values were measured in the freshly prepared solutions before distribution into the replicates. The pH-values of the aged solution were measured from pooled replicates per concentration and control. The temperature of the medium in a surrogate vessel held under the same conditions in the growth room was recorded daily. The light intensity was measured prior to the start of the exposure at positions which had the same distance from the light source as the Lemna fronds. - Reference substance (positive control):
- yes
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 7 d
- Dose descriptor:
- LOEC
- Effect conc.:
- 31.6 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Frond number and dry weight
- Remarks on result:
- other: Growth Rate and Inhibition of Yield
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Frond number and dry weight
- Remarks on result:
- other: Growth Rate and Inhibition of Yield
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: dry weight and frond number yield and growth rate inhibition
- Details on results:
- The environmental conditions (pH-value, room temperature, light intensity) were determined to be within the acceptable limits.
- Results with reference substance (positive control):
- The acute toxicity of 3,5-Dichlorophenol (SIGMA-ALDRICH, batch number MKBZ0947V, purity 100.0 area %, CAS RN 591-35-5) to the monocotyledonous aquatic plant Lemna gibba was determined over a period of 7 days with exposure phase from 2018-04-04 to 2018-04-11 according to OECD Guideline 221. The plants used in the reference test were taken from the same laboratory culture as was used to determine the effects of Basic Red 23.
EC50-Values of the Reference Item based on the nominal concentrations [mg/L], (0-7 days)
Current Study Valid Range (average ± 3 x SD)
Growth rate inhibition (number of fronds)
ErC50 7.68 5.82 ± 3.18
95% confidence interval 7.31 to > 8.00
Yield inhibition (number of fronds)
EyC50 4.93 4.67 ± 2.87
95% confidence interval 4.42 to 5.53
Growth rate inhibition (dry weight)
ErdwC50 > 8.00 5.61 ± 2.76
95% confidence interval Not applicable
Yield inhibition (dry weight)
EydwC50 5.95 4.75 ± 2.49
95% confidence interval 5.18 to 6.75
SD = standard deviation
The observed responses to the reference item were within the valid range, confirming the normal sensitivity of the test system used in the study with the test item. - Reported statistics and error estimates:
- Statistics
For the determination of NOEC, LOEC and EC-values, three replicates were included for the test concentrations and six replicates for the control.
NOEC and LOEC values
NOEC/LOEC was determined by calculation of the statistical significance of inhibition of growth rates and yield in comparison to the control: One Way Analysis of Variance (ANOVA) and DUNNETT’s test were used as a standard. A normality test and an equal variance test were done first. The SHAPIRO-WILK-Test was used to test for normally distributed populations. P-values for both normality and equal variance test were 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) was =0.05.
EC-values and statistical analyses
EC10-, EC20- and EC50-values (0 - 7 d) of the growth rate and yield (frond number and dry weight) inhibition were calculated by sigmoidal dose-response regression. Calculations of the confidence intervals of EC10-, EC20- and EC50-values were carried out from the best fit values, the standard error and the t-distribution with the software GraphPad Prism.
Software
The data for the tables in this report were computer-generated and rounded for presentation from the fully derived data. Consequently, if calculated manually based on the given data, minor deviations may occur from these figures.
Calculations were carried out using the following software:
- Excel, MICROSOFT CORPORATION
- SigmaPlot, SPSS INC.
- GraphPad Prism, GRAPHPAD SOFTWARE, INC.
Any other information on results incl. tables
Frond Numbers
Nominal test item concentration |
Repl. No. |
Frond numbers per study day |
|||
0 days* |
3 days |
5 days |
7 days |
||
100 |
1 |
12 |
20 |
59 |
115 |
2 |
12 |
21 |
59 |
115 |
|
3 |
12 |
22 |
67 |
120 |
|
Mean |
12 |
21 |
62 |
117 |
|
31.6 |
1 |
12 |
20 |
70 |
152 |
2 |
12 |
19 |
53 |
102 |
|
3 |
12 |
20 |
71 |
129 |
|
Mean |
12 |
20 |
65 |
128 |
|
10.0 |
1 |
12 |
19 |
65 |
134 |
2 |
12 |
21 |
70 |
154 |
|
3 |
12 |
23 |
82 |
177 |
|
Mean |
12 |
21 |
72 |
155 |
|
3.16 |
1 |
12 |
19 |
73 |
171 |
2 |
12 |
21 |
83 |
194 |
|
3 |
12 |
21 |
72 |
168 |
|
Mean |
12 |
20 |
76 |
178 |
|
1.00 |
1 |
12 |
22 |
69 |
178 |
2 |
12 |
20 |
82 |
192 |
|
3 |
12 |
21 |
75 |
173 |
|
Mean |
12 |
21 |
75 |
181 |
|
Control |
1 |
12 |
19 |
65 |
157 |
2 |
12 |
22 |
80 |
182 |
|
3 |
12 |
25 |
84 |
206 |
|
4 |
12 |
21 |
75 |
203 |
|
5 |
12 |
19 |
72 |
179 |
|
6 |
12 |
20 |
73 |
185 |
|
Mean |
12 |
21 |
75 |
185 |
* = 3 colonies with 4 fronds each per replicate were inoculated at start of the exposure
Repl.
No. = replicate number
Growth Rate and Yield Inhibition based on Fronds after 7 d
Statistically significant differences of growth rates and yield
compared to control values are marked (+) and non-significant differences are marked (-).
Nominal test item concentration |
Repl. No. |
Average growth rate |
Inhibition of average growth rate |
Yield |
Inhibition of yield |
Doubling time |
|||||||||||
100 |
1 |
|
0.323 |
17 |
|
103 |
41 |
2.15 |
|||||||||
2 |
|
0.323 |
17 |
|
103 |
41 |
2.15 |
||||||||||
3 |
|
0.329 |
16 |
|
108 |
38 |
2.11 |
||||||||||
Mean |
(+) |
0.325 |
17 |
(+) |
105 |
40 |
2.13 |
||||||||||
31.6 |
1 |
|
0.363 |
7 |
|
140 |
19 |
1.91 |
|||||||||
2 |
|
0.306 |
22 |
|
90 |
48 |
2.27 |
||||||||||
3 |
|
0.339 |
13 |
|
117 |
32 |
2.04 |
||||||||||
Mean |
(+) |
0.336 |
14 |
(+) |
116 |
33 |
2.07 |
||||||||||
10.0 |
1 |
|
0.345 |
12 |
|
122 |
30 |
2.01 |
|||||||||
2 |
|
0.365 |
7 |
|
142 |
18 |
1.90 |
||||||||||
3 |
|
0.384 |
1 |
|
165 |
5 |
1.80 |
||||||||||
Mean |
(-) |
0.365 |
7 |
(-) |
143 |
17 |
1.90 |
||||||||||
3.16 |
1 |
|
0.380 |
3 |
|
159 |
8 |
1.83 |
|||||||||
2 |
|
0.398 |
-2 |
|
182 |
-5 |
1.74 |
||||||||||
3 |
|
0.377 |
3 |
|
156 |
10 |
1.84 |
||||||||||
Mean |
(-) |
0.385 |
1 |
(-) |
166 |
4 |
1.80 |
||||||||||
1.00 |
1 |
|
0.385 |
1 |
|
166 |
4 |
1.80 |
|||||||||
2 |
|
0.396 |
-2 |
|
180 |
-4 |
1.75 |
||||||||||
3 |
|
0.381 |
2 |
|
161 |
7 |
1.82 |
||||||||||
Mean |
(-) |
0.388 |
1 |
(-) |
169 |
2 |
1.79 |
||||||||||
Control |
1 |
|
0.367 |
|
|
145 |
|
1.89 |
|||||||||
2 |
|
0.388 |
|
|
170 |
|
1.78 |
||||||||||
3 |
|
0.406 |
|
|
194 |
|
1.71 |
||||||||||
4 |
|
0.404 |
|
|
191 |
|
1.72 |
||||||||||
5 |
|
0.386 |
|
|
167 |
|
1.80 |
||||||||||
6 |
|
0.391 |
|
|
173 |
|
1.77 |
||||||||||
Mean |
|
0.390 |
|
|
173 |
|
1.78 |
Repl. No. = replicate number
Specific Growth Rate and Yield Inhibition of Dry Weight after 7 d
Statistically significant differences of specific growth rates and yield compared
to control values are marked (+) and non-significant differences are marked (-).
Nominal test item concentration |
Repl. No. |
Dry weight |
Specific dry weight growth rate |
Inhibition of specific dry weight growth rate |
Yield of dry weight |
Inhibition of yield dry weight |
|||||||||||
100 |
1 |
18.2 |
|
0.323 |
17 |
|
16.3 |
39 |
|||||||||
2 |
19.0 |
|
0.329 |
15 |
|
17.1 |
36 |
||||||||||
3 |
18.7 |
|
0.327 |
16 |
|
16.8 |
38 |
||||||||||
Mean |
18.6 |
(+) |
0.326 |
16 |
(+) |
16.7 |
38 |
||||||||||
31.6 |
1 |
24.5 |
|
0.365 |
6 |
|
22.6 |
16 |
|||||||||
2 |
17.7 |
|
0.319 |
18 |
|
15.8 |
41 |
||||||||||
3 |
22.8 |
|
0.355 |
9 |
|
20.9 |
22 |
||||||||||
Mean |
21.7 |
(+) |
0.346 |
11 |
(+) |
19.8 |
27 |
||||||||||
10.0 |
1 |
22.5 |
|
0.353 |
9 |
|
20.6 |
23 |
|||||||||
2 |
23.5 |
|
0.359 |
7 |
|
21.6 |
20 |
||||||||||
3 |
28.9 |
|
0.389 |
0 |
|
27.0 |
0 |
||||||||||
Mean |
25.0 |
(-) |
0.367 |
5 |
(-) |
23.1 |
14 |
||||||||||
3.16 |
1 |
26.2 |
|
0.375 |
3 |
|
24.3 |
10 |
|||||||||
2 |
29.2 |
|
0.390 |
-1 |
|
27.3 |
-1 |
||||||||||
3 |
26.5 |
|
0.376 |
3 |
|
24.6 |
9 |
||||||||||
Mean |
27.3 |
(-) |
0.381 |
2 |
(-) |
25.4 |
6 |
||||||||||
1.00 |
1 |
27.9 |
|
0.384 |
1 |
|
26.0 |
3 |
|||||||||
2 |
28.4 |
|
0.386 |
0 |
|
26.5 |
1 |
||||||||||
3 |
26.7 |
|
0.378 |
3 |
|
24.8 |
8 |
||||||||||
Mean |
27.7 |
(-) |
0.383 |
1 |
(-) |
25.8 |
4 |
||||||||||
Control |
1 |
24.9 |
|
0.368 |
|
|
23.0 |
|
|||||||||
2 |
31.1 |
|
0.399 |
|
|
29.2 |
|
||||||||||
3 |
31.1 |
|
0.399 |
|
|
29.2 |
|
||||||||||
4 |
28.8 |
|
0.388 |
|
|
26.9 |
|
||||||||||
5 |
27.8 |
|
0.383 |
|
|
25.9 |
|
||||||||||
6 |
29.1 |
|
0.390 |
|
|
27.2 |
|
||||||||||
Mean |
28.8 |
|
0.388 |
|
|
26.9 |
|
The initial biomass dry weight was 1.9 mg per replicate.
Repl. No. = replicate number
Colony Number (Plants) on Days 0 and 7
Nominal test item concentration |
Replicate No. |
Colony number |
|||||
Day 0 |
Day 7 |
||||||
100 |
1 |
3 |
17 |
||||
2 |
3 |
18 |
|||||
3 |
3 |
17 |
|||||
Mean |
3 |
17 |
|||||
31.6 |
1 |
3 |
20 |
||||
2 |
3 |
13 |
|||||
3 |
3 |
17 |
|||||
Mean |
3 |
17 |
|||||
10.0 |
1 |
3 |
18 |
||||
2 |
3 |
18 |
|||||
3 |
3 |
22 |
|||||
Mean |
3 |
19 |
|||||
3.16 |
1 |
3 |
18 |
||||
2 |
3 |
22 |
|||||
3 |
3 |
18 |
|||||
Mean |
3 |
19 |
|||||
1.00 |
1 |
3 |
19 |
||||
2 |
3 |
19 |
|||||
3 |
3 |
18 |
|||||
Mean |
3 |
19 |
|||||
Control |
1 |
3 |
18 |
||||
2 |
3 |
19 |
|||||
3 |
3 |
24 |
|||||
4 |
3 |
22 |
|||||
5 |
3 |
19 |
|||||
6 |
3 |
18 |
|||||
Mean |
3 |
20 |
Further Observations on Days 2, 5 and 7
Nominal test item concentration |
Observations on day |
||
2 |
5 |
7 |
|
100 |
1 |
1 |
3.3 + |
31.6 |
1 |
1 |
3.3 + |
10.0 |
1 |
1 |
1 |
3.16 |
1 |
1 |
1 |
1.00 |
1 |
1 |
1 |
Control |
1 |
1 |
1 |
Observations were made compared to the appearance of control colonies (plants) and test media
1 = no observedeffects
3.3 = discoloration of roots
+ = slight effects
++ = medium effects
+++ = strong effects
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- In this study, Reactive Blue 71 was found to inhibit the growth of the monocotyledonous aquatic plant Lemna gibba after 7-day exposure under static conditions, with the following effect values (nominal test item concentrations): The EC50-values for both inhibition of the specific growth rate (fronds and dry weight) (ErC50, ErdwC50) were > 100 and yield (fronds and dry weight) (EyC50, EydwC50) were > 100.
- Executive summary:
The effects of the test item Reactive Blue 71 on the growth of the monocotyledonous aquatic plant species Lemna gibba was determined according to the principles of OECD 221 and Council Regulation (EC) No. 761/2009 Method C. 26 at the test facilityfrom2018-06-20 to 2018-06-28, with the definitive exposure phase from 2018-06-20 to 2018-06-27.
Lemna gibbawas exposed to the test item for 7 days under static conditions. Based on a preliminary test, 5 nominal test item concentration levels were tested in a geometrical series with a dilution factor of√10:1.00 - 3.16 - 10.0 - 31.6 - 100 mg/L. Three replicates were investigated for each test concentration and six for the control. Frond numbers were assessed on days 0, 2, 5 and 7. Environmental parameters (light, pH and temperature) were within the acceptable limits. All test solutions were clear and concentration-related blue coloured throughout the exposure.The validity criteria of the test guideline were fulfilled.
The concentrations ofthe test itemReactive Blue71and the controlwere analysed via HPLC-DAD at the first interval. The control, the lowest and the highest test item concentration were analytically verified at the end of the exposure and on every renewal day. Details of the analytical method are presented in section13.
At the start of the first interval the measured concentrations ofReactive Blue 71were in the range of 101 and 107% of the nominal values. At the end of the first interval the measured concentrations were between 98 and 105% of the nominal values. For details of the analytical results and method, please refer to section8.1.3. All effect values are given based on the nominal test item concentrations(Table 1).
Table1: NOEC-, LOEC-, EC-Values and 95% Confidence Intervals ofReactive Blue 71after 7 Days of Exposure
(based on the nominal test item concentration [mg/L])
Frond number
Dry weight
Growth Rate Inhibition [mg/L]
NOEC
10.0
10.0
LOEC
31.6
31.6
ErC10
16.4 (8.04 – 36.6)
27.1 (12.5 – 61.0)
ErC20
> 100
> 100
ErC50
> 100
> 100
Inhibition of Yield [mg/L]
NOEC
10.0
10.0
LOEC
31.6
31.6
EyC10
5.92 (<1.00 – 12.5)
6.33 (<1.00 – 16.2)
EyC20
12.0 (5.45 – 26.6)
18.0 (6.97 – 43.2)
EyC50
> 100
> 100
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