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Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-11-22 to 2018-12-13
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 208 (Terrestrial Plants, Growth Test)
Version / remarks:
July 2006
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Details on sampling:
Determination of the test item
The concentration of the test item EDTA-CuK2 was confirmed by analytical verification of the stock solution and control.
The samples were analysed with a photometer method, implemented under non-GLP and documented finally in the GLP raw data.
Vehicle:
no
Details on preparation and application of test substrate:
The control and test item treatments were applied once at the beginning of the study. A stock solution of 60 g/L was prepared in demineralised water, diluted to various concentrations and incorporated into the soil. All concentrations of the test item were related to the dry weight of the soil. The test item solutions and additional demineralised water were added to the soil to adjust the humidity of the soil to a moisture of 45 % of the maximum water holding capacity (MWHC) were added to the soil. Subsequently, the soil was mixed carefully with a mixer (3 min) to ensure a homogeneous distribution.
Due to large soil amounts per concentration, several batches of 7.8 kg soil dry weight were prepared per concentration and control and applied separately.
Total soil amount:
Each test item concentration and control: 15.6 kg DW

Application Conditions of the Soil
Application date 2018-11-22
Dry Weight (DW) of the soil before application [g/100 g soil WW] 96.2
Soil amount per batch [kg DW] 7.80
[kg WW] 8.11
Maximum water holding capacity (MWHC) of the soil [g/100 g soil DW] 37.0
Added water amount per application to reach 45 % of MWHC [g/ 100 g DW] 12.7
Soil amount (with 45 % of MWHC) per replicate [kg] 0.700

The treated soil was transferred to the test container and the seeds were sown.
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
Common name: Oats
Plant family: Poaceae
Variety: KWS Contender
Source of seed: KWS Lochow GmbH, Ferdinand-von-Lochow-Straße 5, 29303 Bergen, Germany
Prior seed treatment / Sterilization: No
Seed storage: The seeds were stored dark at the test facility at room temperature (22 +/- 2 °C) and protected from moisture until use.
Species:
Brassica napus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Common name: Rape
Plant family: Brassicaceae
Variety: Sherlock
Source of seed: KWS Saat SE, Grimsehlstraße 31, 37574 Einbeck, Germany
Prior seed treatment / Sterilization: No
Seed storage: The seeds were stored dark at the test facility at room temperature (22 +/- 2 °C) and protected from moisture until use.
Species:
Glycine max (G. soja)
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Common name: Soybean
Plant family: Fabaceae
Variety: Obelix
Source of seed: Delley Samen und Pflanzen AG, Schloss Delley 40, route de Portalban, CH-1567 Delley, Switzerland
Prior seed treatment / Sterilization: No
Seed storage: The seeds were stored dark at the test facility at room temperature (22 +/- 2 °C) and protected from moisture until use.
Test type:
seedling emergence and seedling growth test
Study type:
laboratory study
Substrate type:
other: 2:1 Mixture of natural soil LUFA 2.2 (loamy sand (DIN classification)) and quartz sand
Limit test:
no
Total exposure duration:
21 d
Test temperature:
nominal: 22 +/- 10 °C; actual: 13.8 - 21.8 °C
pH:
5.36 ± 0.04 (soil)
Moisture:
relative humidity:
nominal: 70 +/- 30 %; 39.0 - 89.2 %
Details on test conditions:
TEST SYSTEM
- Testing facility: Noack Laboratorien GmbH
- Test container (type, material, size): Non-porous plastic containers (standard flower pots) with a diameter of ca. 12 cm were used. The pots were labelled with the study number and all necessary additional information to ensure unique identification.
- Amount of soil: 700 g per test container
- Method of seeding: Test containers were filled each with the same amount of treated soil (see application technique) and seeding holes were made with a seedling pistil. One seed was given into each hole and covered with soil. During cultivation the test containers were bottom watered and fertilized as necessary.
- No. of seeds per container: Oats, rape: 5; Soybean: 3
- No. of replicates per treatment group: 8
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
A 2:1 mixture of natural soil LUFA 2.2 (batch number: Sp2.23618, loamy sand (DIN classification)) and quartz sand (12a) was used.
Grain size: <= 2 mm
Clay (< 0.002 mm): 6.0 %
Silt (0.002 - 0.063 mm): 9.1 %
Sand (0.063 - 2 mm): 84.9 %
Carbon content: 0.986 %
pH-value: 5.36 ± 0.04

NUTRIENT MEDIUM (if used)
HAKAPHOS® SOFT SPEZIAL 16+8+22(+3)
For watering a 0.05 ‰ solution was used
Origin: Grünes Landhaus, St. Godehard-Str. 23, 31139 Hildesheim, Germany

GROWTH CONDITIONS
- Photoperiod: 16/8
- Light source: Special-fluorescent tube Fluora and Daylight (Osram)
- Light intensity and quality: nominal: > 3500 lx; actual: 4570 ± 491
- Temperatures: nominal: 22 +/- 10 °C; 13.8 - 21.8 °C
- Relative humidity (%): nominal: 70 +/- 30 %; actual: 39.0 - 89.2 %
- Watering regime and schedules: Bottom watering of the test containers with fertilized tap water as needed.
- Interval of applications: once at experimental starting
- Method of application: spray application

ACCLIMATION PERIOD: None

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

- Phytotoxicity rating system (if used):
During the 21 day observation period (from day of application) the plants were observed on day 7, 14 and 21, for number of emerged seedlings, number of dead plants and visual phytotoxic effects.
The rating of the treated plants was done in relation to the untreated control plants. Observations included all variations, either inhibitory or stimulatory, between the treated replicates and the untreated controls. Such variations may be phytotoxic symptoms (e.g. chlorosis, necrosis, wilting), effects of growth and development rates. 
Observation Parameters
(1) Deformation of leaves (curling / twisting / sticking / …)
(2) Shoot deformations (curling / bending / …)
(3) Stunted growth
(4) Necrosis
(5) Chlorosis
(6) Lighter green
(7) Darker green
(8) Loss of colour
(9) Discoloration of leaves
(10) Wilting
(E) Dead plant (Number of dead plants)

Assessment of the Effects
- = no effects observed / normal growth
+/- = slight effects
+ = medium effects
++ = strong effects 


VEHICLE CONTROL PERFORMED: yes/no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Range finding study
The preliminary study was performed under the same laboratory conditions as specified for the definitive study with 4 replicates per concentration and control. Test duration was 21 days. At test end shoot fresh weight and emergence were determined.

Inhibition of Shoot Fresh Weight
Test concentration
[mg/kg DW] Inhibition [%]
Oats Rape Soybean
10 -5 -4 -9
100 0 6 3
1000 29 87 -

Inhibition of Rate of Emergence
Test concentration
[mg/kg DW] Inhibition [%]
Oats Rape Soybean
10 10 0 0
100 10 0 11
1000 10 55 100
Negative values = promoted growth / promoted emergence
DW = soil dry weight
- not emerged
Nominal and measured concentrations:
Nominal: 1000 - 400 - 160 - 64 - 25.6 mg/kg DW
Reference substance (positive control):
no
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoor fresh weight
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
964 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
CI (95%) = 746 - > 1000 mg/kg soil DW
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
160 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
490 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Remarks on result:
other:
Remarks:
CI (95%) = 364 - 626 mg/kg soil DW
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
160 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
230 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
CI (95 %) = 182 - 300 mg/kg sol DW
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
403 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
CI (95%) = 317 - 500 mg/kg soil DW
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
462 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
CI (95%) = 354 - 579 mg/kg soil DW
Key result
Species:
Brassica napus
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
705 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
CI (95%) = 578 - 851 mg/kg soil DW
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot height dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot height dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot height dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot height
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot height dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot fresh weight dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot fresh weight dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot fresh weight dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth
Remarks:
shoot fresh weight
Remarks on result:
other:
Remarks:
1000 mg/kg DW: no germination. No shoot fresh weight dertermination possible.
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
160 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
400 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC25
Effect conc.:
49.9 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
CI (95%) = < 25.6 - 202 mg/kg
Key result
Species:
Glycine max (G. soja)
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
225 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other:
Remarks:
CI (95%) = 107 - 410 mg/kg soil DW
Details on results:
The percentage of inhibition of shoot height, shoot fresh weight and rate of emergence is given ibelow. Test item-related statistically significant inhibitions are given in bold. The tabulated results represent rounded mean values calculated on the exact raw data.

Oats: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence
Test concentration
[mg/kg DW] Shoot
height1)
[cm] Inhibition

[%] Shoot fresh weight
[mg] Inhibition

[%] Rate of emergence1)
[%] Inhibition

[%]
Control 38.9 - 989 - 98 -
25.6 38.5 1 1125 -14 85 13
64 41.1 -6 1179 -19 98 0
160 39.6 -2 1033 -4 100 -2
400 39.3 -1 1042 -5 93 5
1000 32.0 18 723 27 90 8


Rape: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence
Test concentration
[mg/kg DW] Shoot
height1)
[cm] Inhibition

[%] Shoot fresh weight
[mg] Inhibition

[%] Rate of emergence1)
[%] Inhibition

[%]
Control 15.0 1278 98
25.6 14.7 2 1158 9 100 -2
64 14.1 6 1113 13 98 0
160 14.3 5 1122 12 93 5
400 11.4 24 627 51 80 18
1000 8.7 42 378 70 28 71

Soybean: Inhibition [%] of Shoot Height, Shoot Fresh Weight and Rate of Emergence
Test concentration
[mg/kg DW] Shoot
height
[cm] Inhibition

[%] Shoot fresh weight1)
[mg] Inhibition

[%] Rate of emergence
[%] Inhibition

[%]
Control 16.4 1959 75
25.6 17.1 -4 1882 4 58 23
64 16.7 -2 1727 12 54 28
160 15.7 4 1710 13 46 39
400 15.3 7 1222 38 21 72
1000 # - # - 0 100
Negative values = Promoted growth
Bold = Statistically significant inhibition
1) Normality test failed
#) For the test concentration 1000 mg/kg DW no seedling emergence occurred. Therefore, no shoot height and
shoot fresh weight measurements were possible.
Results with reference substance (positive control):
None
Reported statistics and error estimates:
Percentage of inhibition
The percentage of inhibition of biomass growth (shoot height, shoot fresh weight) and rate of emergence were calculated in relation to the control. All dead plants were excluded from the calculation of shoot height and fresh weight (above-ground biomass).

NOEC / LOEC of biomass growth / emergence
One Way Analysis of Variance (ANOVA) was carried out for the determination of statistically significant differences compared to control replicates. When running a One Way Analysis of Variance, a Normality test (Shapiro-Wilk) and an Equal Variance test (Brown-Forsythe) were done first. P-values for both, Normality and Equal Variance test, are 0.05. The alpha-value for ANOVA (acceptable probability of incorrectly concluding that there is a difference) is alpha = 0.05.

Failure of the normality test and / or the equal variance test can be caused by extremely homogeneous emergence and growth patterns as opposed to higher variances in other treatments. Due to the high and even number of replicates in the control and treatment groups, the failure had no influence on the robustness of the calculations.

EC-values and confidence intervals of biomass growth
EC25- and EC50- values were calculated by sigmoidal dose-response regression using GraphPadPrism. Four parameter were used for dose-response fitting. Calculation of the confidence intervals for EC-values was carried out using standard procedures provided by GraphPadPrism.
Validity criteria fulfilled:
yes
Conclusions:
Potential toxic effects of the test item were assessed on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and at test end (day 21) by shoot height and shoot fresh weight determination.
The results of the control for all plant species met the required validity criteria. Therefore, the test is considered to be valid.

Shoot Height: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)
Species NOEC
[mg/kg DW] LOEC
[mg/kg DW] EC25
[mg/kg DW] p = 95%
[mg/kg DW] EC50
[mg/kg DW] p = 95%
[mg/kg DW]
Oats 400 1000 > 1000 ̶ > 1000 ̶
Rape 160 400 490 364 - 626 > 1000 ̶
Soybean ≥ 400# > 400# > 400# ̶ > 400# ̶


Shoot Fresh Weight: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)
Species NOEC
[mg/kg DW] LOEC
[mg/kg DW] EC25
[mg/kg DW] p = 95%
[mg/kg DW] EC50
[mg/kg DW] p = 95%
[mg/kg DW]
Oats 400 1000 964 746 - > 1000 > 1000 ̶
Rape 160 400 230 182 - 300 403 317 - 500
Soybean ≥ 400# > 400# 279 120 - > 400# > 400# ̶


Number of Emerged Seedlings: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)
Species NOEC
[mg/kg DW] LOEC
[mg/kg DW] EC25
[mg/kg DW] p = 95%
[mg/kg DW] EC50
[mg/kg DW] p = 95%
[mg/kg DW]
Oats ≥ 1000 > 1000 > 1000 ̶ > 1000 ̶
Rape 400 1000 462 354 - 579 705 578 - 851
Soybean 160 400 49.9 < 25.6 - 202 225 107 - 410
-) Not determinable
#) For the test concentration 1000 mg/kg DW no seedling emergence occurred. Therefore, no shoot height and
shoot fresh weight measurements were possible.
Executive summary:

Summary

The phytotoxicity of EDTA-CuK2 to three terrestrial plant species was determined according to OECD 208 (July 2006) in a seedling emergence and seedling growth tests over a period of 21 days.

Test Design

Test systems comprised one monocotyledon (oats, Poaceae) and two dicotyledons (rape, Brassicaceae; soybean, Fabaceae).

The test was conducted with the test concentrations of1000 - 400 - 160 - 64 - 25.6 mg/kg soil dry weight (DW) (factor 2.5).

The test item was incorporated into the soil in which the seeds were sown. The test containers were bottom watered with nutrient solution as needed.

 

Results

The stock solution was sampled prior to application and subsequently analytically verified. The measured concentration of EDTA-CuK2 was 101 % indicating the correct preparation of the stock solution.

Potential toxic effects of the test item were assessed on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and at test end by shoot height and shoot fresh weight determination.

 

The results of the control for all plant species met the required validity criteria. Therefore, the test is considered to be valid.

 

Shoot Height: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

400

1000

> 1000

̶

> 1000

̶

Rape

160

400

490

364 - 626

> 1000

̶

Soybean

≥ 400#

> 400#

> 400#

̶

> 400#

̶

 

Shoot Fresh Weight: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

400

1000

964

746 - > 1000

> 1000

̶

Rape

160

400

230

182 - 300

403

317 - 500

Soybean

≥ 400#

> 400#

279

120 - > 400#

> 400#

̶

 

 Number of Emerged Seedlings: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

≥ 1000

> 1000

> 1000

̶

> 1000

̶

Rape

400

1000

462

354 - 579

705

578 - 851

Soybean

160

400

49.9

< 25.6 - 202

225

107 - 410

-) Not determinable
#) For the test concentration 1000 mg/kg DW no seedling emergence occurred. Therefore, no shoot height and
   shoot fresh weight measurements were possible.

Description of key information

One toxicity to terrestrial plant tests is performed with EDTA-CuK2 according to OECD 208 using three different plant species. the lowest NOEC of 160 mg test item/kg dw was observed for the growth, shootheight and shoot wet weight of Brassica napus (rape). The lowest EC50 of 225 mg test item/kg dw was observed for the number of emerged seedlings for Glycine max (soybean). The test item contained 92% active ingredient. The effect data based on active are then: NOEC of 147.2 mg a.i./kg dw was observed for the growth, shootheight and shoot wet weight of Brassica napus (rape). The lowest EC50 of 207 mg a.i./kg dw

Key value for chemical safety assessment

Short-term EC50 or LC50 for terrestrial plants:
207 mg/kg soil dw
Long-term EC10, LC10 or NOEC for terrestrial plants:
147.2 mg/kg soil dw

Additional information

The phytotoxicity of EDTA-CuK2 to three terrestrial plant species was determined according to OECD 208 (July 2006) in a seedling emergence and seedling growth tests over a period of 21 days.

 

The test systems comprised one monocotyledon (oats, Poaceae) and two dicotyledons (rape, Brassicaceae; soybean, Fabaceae). 

The test was conducted with the test concentrations of 1000 - 400 - 160 - 64 - 25.6 mg/kg soil dry weight (DW) (factor 2.5).

 The test item dissolves well in water and was incorporated into the soil in which the seeds were sown. The test containers were bottom watered with nutrient solution as needed.

 

The stock solution was sampled prior to application and subsequently analytically verified. The measured concentration of EDTA-CuK2was 101 % indicating the correct preparation of the stock solution.

 

Potential toxic effects of the test item were assessed on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and at test end by shoot height and shoot fresh weight determination.

 

The results of the control for all plant species met the required validity criteria. Therefore, the test is considered to be valid.


 

Table1:          Shoot Height: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

400

1000

> 1000

̶

> 1000

̶

Rape

160

400

490

364 - 626

> 1000

̶

Soybean

≥ 400#

> 400#

> 400#

̶

> 400#

̶


Table2:          Shoot Fresh Weight: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

400

1000

964

746 - > 1000

> 1000

̶

Rape

160

400

230

182 - 300

403

317 - 500

Soybean

≥ 400#

> 400#

279

120 - > 400#

> 400#

̶

 

 

Table3:          Number of Emerged Seedlings: NOEC, LOEC, EC25- and EC50-Values with Confidence Range (p)

Species

NOEC
[mg/kg DW]

LOEC
[mg/kg DW]

EC25
[mg/kg DW]

p = 95%
[mg/kg DW]

EC50
[mg/kg DW]

p = 95%
[mg/kg DW]

Oats

≥ 1000

> 1000

> 1000

̶

> 1000

̶

Rape

400

1000

462

354 - 579

705

578 - 851

Soybean

160

400

49.9

< 25.6 - 202

225

107 - 410

-)Not determinable
#) For the test concentration 1000 mg/kg DW no seedling emergence occurred. Therefore, no shoot height and
   shoot fresh weight measurements were possible.