Registration Dossier

Ecotoxicological information

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 2018-07-25 to 2018-08-02, with the definitive exposure phase from 2018-07-25 to 2018-08-01
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
Version / remarks:
2006
Deviations:
no
Qualifier:
according to
Guideline:
other: Council Regulation (EC) No. 761/2009 Method C.26
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

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 start (0 day, fresh medium) and at the end of the exposure (7 days, old medium). The samples were analysed with an HPLC-DAD method. The method was implemented under non-GLP but documented in the raw data and validated according to Annex I. 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. The test item was weighed out on a glass side and was inserted in a glass flask which contains the dilution water. This stock solution was stirred for one hour at room temperature at 1100 rpm.

Test concentrations
Based on the results of a preliminary range finding test , 5 concentrations were tested with a dilution factor of 4: 0.0391 - 0.156 - 0.625 - 2.50 - 10.0 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 Concentration in prepared medium
[g/L] [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:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
7 d

Test conditions

Hardness:
not measured
Test temperature:
Room temperature [°C]: min.: 23.3 max.: 24.1 mean value: 22.7
pH:
Nominal
test item concentration pH-value
[mg/L] 0 h (Fresh medium) 7 d (Old medium)
10.0 7.72 7.99
2.50 7.72 7.99
0.625 7.71 8.00
0.156 7.70 8.01
0.0391 7.70 8.01
Control 7.66 8.00
Dissolved oxygen:
not measured
Salinity:
not measured
Conductivity:
not measured
Nominal and measured concentrations:
Nominal test item concentration: 0.0391 - 0.156 - 0.625 - 2.50 - 10.0 mg/L
Geometric mean measured test item concentration: 0.300 - 0.300 - 0.300 - 1.34 - 7.79 mg/L
Details on test conditions:
Test method
Static 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
Static with application of the test item at test start. 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:
NOEC
Effect conc.:
0.039 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:
LOEC
Effect conc.:
0.156 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.:
0.803 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number growth rate inhibition
Remarks on result:
other: 95% Confidence limits: 0.600 - 1.13
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
0.095 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number growth rate inhibition
Remarks on result:
other: 95% Confidence limits: 0.0660 - 0.139
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
0.17 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number growth rate inhibition
Remarks on result:
other: 95% Confidence limits: 0.120 - 0.235
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
0.118 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight growth inhibition
Remarks on result:
other: 95% Confidence limits: 0.0902 - 0.155
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
0.211 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight growth inhibition
Remarks on result:
other: 95% Confidence limits: 0.166 - 0.264
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
0.862 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight growth inhibition
Remarks on result:
other: 95% Confidence limits: 0.711 - 1.07
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
0.073 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.0547 - 0.0953
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
0.1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.0782 - 0.125
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
0.215 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: frond number inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.179 - 0.262
Duration:
7 d
Dose descriptor:
EC10
Effect conc.:
0.079 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.0622 - 0.0993
Duration:
7 d
Dose descriptor:
EC20
Effect conc.:
0.108 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.0869 - 0.132
Duration:
7 d
Dose descriptor:
EC50
Effect conc.:
0.238 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight inhibition of yield
Remarks on result:
other: 95% Confidence limits: 0.201 - 0.285
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 from 2018-04-04 to 2018-04-12, with the definitive exposure phase from 2018-04-04 to 2018-04-11 according to OECD Guideline 221.

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 – > 8.00
Yield inhibition (number of fronds)
EyC50 4.93 4.67 ± 2.87
95% confidence interval 4.42 – 5.53
Growth rate inhibition (dry weight)
ErdwC50 > 8.00 5.61 ± 2.76
95% confidence interval -
Yield inhibition (dry weight)
EydwC50 5.95 4.75 ± 2.49
95% confidence interval 5.18 – 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. 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
[mg/L]

Repl.

No.

Frond numbers per study day

0 days*

2 days

5 days

7 days

10.0

1

12

15

24

21

2

12

18

23

22

3

12

15

22

19

Mean

12

16

23

21

 2.50

1

12

20

30

37

2

12

21

33

38

3

12

19

30

35

Mean

12

20

31

37

 0.625

1

12

21

47

53

2

12

23

48

55

3

12

23

41

46

Mean

12

22

45

51

 0.165

1

12

22

79

126

2

12

22

87

145

3

12

24

89

175

Mean

12

23

85

149

 0.0391

1

12

27

124

248

2

12

23

106

227

3

12

26

95

228

Mean

12

25

108

234

Control

1

12

24

107

237

2

12

22

96

245

3

12

23

95

222

4

12

21

94

207

5

12

24

100

234

6

12

24

104

226

Mean

12

23

99

229

* = 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
[mg/L]

Repl.

No.

Average growth rate

[d-1]

Inhibition of average growth rate
[%]

Yield


[fronds]

Inhibition of yield

[%]

Doubling time

[d]

10.0

1

 

0.0799

81

 

9

96

8.67

2

 

0.0866

79

 

10

95

8.00

3

 

0.0656

84

 

7

97

10.6

Mean

(+)

0.0770

82

(+)

9

96

9.08

 2.50

1

 

0.161

62

 

25

88

4.31

2

 

0.165

61

 

26

88

4.21

3

 

0.153

64

 

23

89

4.53

Mean

(+)

0.159

62

(+)

25

89

4.35

 0.625

1

 

0.212

50

 

41

81

3.27

2

 

0.217

48

 

43

80

3.19

3

 

0.192

54

 

34

84

3.61

Mean

(+)

0.207

51

(+)

39

82

3.35

 0.165

1

 

0.336

20

 

114

47

2.06

2

 

0.356

15

 

133

39

1.95

3

 

0.383

9

 

163

25

1.81

Mean

(+)

0.358

15

(+)

137

37

1.94

 0.0391

1

 

0.433

-3

 

236

-9

1.60

2

 

0.420

0

 

215

1

1.65

3

 

0.421

0

 

216

0

1.65

Mean

(-)

0.424

-1

(-)

222

-3

1.63

Control

1

 

0.426

 

 

225

 

1.63

2

 

0.431

 

 

233

 

1.61

3

 

0.417

 

 

210

 

1.66

4

 

0.407

 

 

195

 

1.70

5

 

0.424

 

 

222

 

1.63

6

 

0.419

 

 

214

 

1.65

Mean

 

0.421

 

 

217

 

1.65

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
[mg/L]

Repl.

No.

Dry weight


[mg]

Specific dry weight

growth rate

[d-1]

Inhibition of specific dry weight growth rate
[%]

Yield of dry weight


[mg]

Inhibition of yield dry weight
 
[%]

10.0

1

3.1

 

0.0777

82

 

1.3

96

2

3.6

 

0.0990

77

 

1.8

95

3

2.8

 

0.0631

86

 

1.0

97

Mean

3.2

(+)

0.0800

82

(+)

1.4

96

 2.50

1

5.0

 

0.146

67

 

3.2

91

2

5.7

 

0.165

62

 

3.9

89

3

4.6

 

0.134

69

 

2.8

92

Mean

5.1

(+)

0.148

66

(+)

3.3

91

 0.625

1

9.4

 

0.236

46

 

7.6

79

2

10.3

 

0.249

43

 

8.5

77

3

8.8

 

0.227

48

 

7.0

81

Mean

9.5

(+)

0.237

46

(+)

7.7

79

 0.165

1

22.5

 

0.361

17

 

20.7

43

2

27.4

 

0.389

11

 

25.6

30

3

28.1

 

0.393

10

 

26.3

28

Mean

26.0

(+)

0.381

13

(+)

24.2

34

 0.0391

1

43.5

 

0.455

-4

 

41.7

-15

2

39.8

 

0.442

-1

 

38.0

-4

3

37.1

 

0.432

1

 

35.3

3

Mean

40.1

(-)

0.443

-2

(-)

38.3

-5

Control

1

38.1

 

0.436

 

 

36.3

 

2

40.3

 

0.444

 

 

38.5

 

3

38.0

 

0.436

 

 

36.2

 

4

36.3

 

0.429

 

 

34.5

 

5

38.8

 

0.439

 

 

37.0

 

6

37.8

 

0.435

 

 

36.0

 

Mean

38.2

 

0.436

 

 

36.4

 

The initial biomass dry weight was 1.8 mg per replicate.

Repl. No. = replicate number

Colony Number (Plants) on Days 0 and 7

 

Nominal test item concentration
[mg/L]

Replicate

No.


Colony number

Day 0

Day 7

10.0

1

3

19

2

3

11

3

3

10

Mean

3

13

 2.50

1

3

22

2

3

23

3

3

20

Mean

3

22

 0.625

1

3

13

2

3

13

3

3

11

Mean

3

12

 0.165

1

3

22

2

3

23

3

3

29

Mean

3

25

 0.0391

1

3

38

2

3

35

3

3

32

Mean

3

35

Control

1

3

31

2

3

29

3

3

28

4

3

23

5

3

36

6

3

29

Mean

3

29

Nominal test item concentration
[mg/L]

Observations on day

2

5

7

10.0

2.4 ++
3.2 ++
3.3 +++
4.1 ++

2.4 ++
2.5 +
3.2 +++
3.3 +++
4.1 ++

2.1 ++
2.4 ++
2.5 ++
3.2 +++
3.3 +++
4.1 ++

 2.50

2.4 +
3.2 +
3.3 ++

2.4 ++
2.5 +
3.2 +++
3.3 ++
4.1 ++

2.1 +
2.4 ++
2.5 +
3.2 +++
3.3 ++
4.1 ++

 0.625

3.3 +

3.2 +
3.3 +

3.2 +
3.3 +

 0.156

1

1

3.2 +

 0.0391

1

1

1

Control

1

1

1

Observations were made compared to the appearance of control colonies (plants) and test media

 

1      = no observedeffects

2.1   = chlorosis

2.4   = discoloration of the fronds

2.5   = smaller frond

3.2  = losses of roots

3.3   = discoloration of roots

4.1   = break up of plants

+      = slight effects

++    = medium effects

+++  = strong effects

The concentrations ofBasicBlue 45Methylsulfatwere determined at the start of the exposure in the fresh media (0 h) and at the end of the exposure in the old media (7 d) of all tested concentration levels and the control via HPLC-DAD.For results seeTable9.Details of the analytical method are presented in part 14.

The measured concentrations ofBasicBlue45 Methylsulfatin the fresh media (0 d) were in the range of < LOQ to 102% and in the old media (7 d) in the range of < LOQ to 58% of nominal test item concentrations. The measured test item concentrations were not within the range of±20% of the nominal concentrations, the geometric mean measured concentrations were calculated.

 

Table9:      Measured Concentrations and Percent of Nominal Concentration of the Test Item
Basic Blue45Methylsulfatduring the Definitive Test

Sampling date

fresh medium

old medium

 

Start of analysis

2018-07-25

2018-08-01

 

Nominal test item concentration

[mg/L]

Basic Blue45Methylsulfat

 

Meas. conc.

[mg/L]

%

Meas. conc.

[mg/L]

%

Geometric mean measured
concentration
[mg/L]2)

10.0

10.2

102

5.93

58

7.79

2.50

2.23

89

0.811

36

1.34

0.625

0.4551)

73

< LOQ

0.300

0.156

< LOQ

< LOQ

0.300

0.0391

< LOQ

< LOQ

0.300

Control

< LOQ

< LOQ

 

Meas. conc.    = measured concentration of the test item, mean value of two injections, dilution factor

  taken into account

%                     = percent of the nominal concentration of the test item

LOQ                = limit of quantification (0.600 mg test item/L)

1)                     = < LOQ, but70% LOQ

2)                     = If the measured concentration was < LOQ, 1/2 LOQ (0.300 mg test item/L) was used for
   the calculation of the
 geometric mean measured concentration.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this study, Basic Blue 45 Methylsulfat 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 inhibition of the specific growth rate of fronds (ErC50) and dry weight (ErdwC50) were 0.803 (0.600 – 1.13) mg/L and 0.862 (0.711 – 1.07 mg/L, respectively. The EC50-values for inhibition of yield (fronds, EyC50) and dry weight inhibition of yield (EydwC50) were 0.215 (0.179 – 0.262) mg/L and 0.238 (0.201 – 0.285) mg/L, respectively. For further values (EC10-values, EC20-values and NOEC/LOEC) of the test item Basic Blue 45 Methylsulfat.
Executive summary:

The effects of the test item Basic Blue 45 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 facility from 2018 -07 -25 to 2018 -08 -02, with the definitive exposure phase from 2018 -07 -25 to 2018 -08 -01.

 

Lemna gibba was 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 4: 0.0391 - 0.156 - 0.625 - 2.50 - 10.0 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.The validity criteria of the test guideline were fulfilled. The test item solutions were clear and concentration related blue coloured throughout the exposure period.

 

The concentrations of the test item Basic Blue45 Methylsulfat and the control were analysed via HPLC-DAD at the beginning and end of the exposure.

The measured concentrations of Basic Blue 45 Methylsulfat in the fresh media (0 d) were in the range of > LOQ to 102% and in the old media (7 d) in the range of < LOQ to 58% of the nominal test item concentrations. All effect values are given based on the nominal test item concentrations.

 

NOEC-, LOEC-, EC-Values and 95% Confidence Intervals ofBasic Blue 45 Methylsulfat after 7 Days of Exposure

(based on the nominal test item concentration [mg/L])

Frond number

Dry weight

Growth Rate Inhibition [mg/L]

NOEC

0.0391

NOEC

0.0391

LOEC

0.156

LOEC

0.156

ErC10

0.0953 (0.0660 – 0.139)

ErdwC10

0.118 (0.0902 – 0.155)

ErC20

0.170 (0.120 – 0.235)

ErdwC20

0.211 (0.166 – 0.264)

ErC50

0.803 (0.600 – 1.13)

ErdwC50

0.862 (0.711 – 1.07)

Inhibition of Yield [mg/L]

NOEC

0.0391

NOEC

0.0391

LOEC

0.156

LOEC

0.156

EyC10

0.0727 (0.0547 – 0.0953)

EydwC10

0.0787 (0.0622 – 0.0993)

EyC20

0.100 (0.0782 – 0.125)

EydwC20

0.108 (0.0869 – 0.132)

EyC50

0.215 (0.179 – 0.262)

EydwC50

0.238 (0.201 – 0.285)