Fatty acids, tall-oil, reaction products with 2-[(2-aminoethyl)amino]ethanol

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Endpoint:

toxicity to terrestrial plants: long-term

Remarks:

Effect on terrestrial (non-target) plants: seedling emergence and seedling growth test

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Dec 2015 - July 2016

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants, Growth Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: 1583127
- Expiration date of the batch: March 04, 2017
- Purity: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature, in the dark

Analytical monitoring:

no

Remarks:

The test substance is a UVCB substance with a water solubility <10 mg/L. No analytical method for quantification of one or more ingredients was available for this UVCB substance in soil.

Details on sampling:

According to OECD 208 it is sufficient for insoluble substances to verify the test concentrations by providing the weights of the test substance added to the soil.

Vehicle:

yes

Remarks:

acetone

Details on preparation and application of test substrate:

The test item was dissolved in acetone and mixed with fine quartz sand before the test item treated sand was mixed into the soil. The amount of quartz sand depended on the water content of the used charge of soil. The amount of quartz sand and acetone per kg soil was the same in all test item treatment groups and the solvent control (2.19 mL acetone / kg dry soil).
A stock solution of 113.9002 g test item ad 250 mL acetone was prepared. For the following concentrations 60 mL out of the next higher concentration were added to 120 mL acetone. For the last two concentrations only 20 mL out of the next higher concentration were taken and 40 mL acetone was added.

Then adequate volumes of the acetone dilutions were given to fine quartz sand and mixed with a laboratory mixer. The acetone was allowed to evaporate overnight out of the sand mixtures under a fume hood. The next day, all sand mixtures were given on adequate amounts of soil and each batch was mixed thoroughly for 10 minutes in a laboratory mixer.
The acetone control was treated with the same amount of acetone and sand as the test item groups. The control was treated with the same amount of untreated sand as the test item groups and acetone control.
The soil for one concentration was treated in one or two batches and then split into the replicates.

Species:

other: Solanum lycopersicon

Plant group:

Dicotyledonae (dicots)

Species:

Helianthus annuus

Plant group:

Dicotyledonae (dicots)

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Species:

Allium cepa

Plant group:

Monocotyledonae (monocots)

Test type:

seedling emergence and seedling growth test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

21 d

Test temperature:

Mean: 21.6 °C
Min: 16.4 °C
Max: 25.6 °C

pH:

5.7 ± 0.6

Moisture:

Mean: 60 %
Min: 49 %
Max: 87 %

Details on test conditions:

TEST SYSTEM
- Testing facility: ibacon GmbH, Rossdorf, Germany
- Test container (type, material, size): commercial plastic flower pots (diameter: 12 cm or 14 cm)
- Amount of soil:
- Method of seeding: the seeds were introduced manually into the test item treated soil
- No. of seeds per container: 5
- No. of replicates per treatment group: 6
- No. of replicates per control: 6
- No. of replicates per vehicle control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
The soil was steam sterilized, delivered and analysed by the LUFA Speyer, Germany.
- Soil Type: LUFA 2.3 (USDA: sandy loam)
- Geographic location: Offenbach, Germany
- Pesticide use history at the collection site: no pesticide treatments were performed in the year of sampling and for at least the two previous years
- Sampling depth (cm): 20
- Soil texture (if natural soil): sandy loam
- Particle size: all particles ≤ 0.2 cm
- Organic carbon (%): 0.67 ± 0.03%
- pH: 5.7 ± 0.6

GROWTH CONDITIONS
The test was performed in a growth chamber under controlled conditions.
- Photoperiod: 16 h light : 8 h dark
- Light intensity: 9,870-19,930 Lux (mean: 13,823 Lux)
- Temperatures (°C): 16.4-25.6 (mean: 21.6 °C)
- Relative humidity (%): 49-87 (mean: 60 %)
- Watering regime and schedules: A spot check on soil moisture was carried out daily. Bottom watering (through saucers) was done where necessary. Water was given in order to assure optimal water supply of the plants.
- Fertilizer: After development of the first true leaves, Ferty® 9 “Hydro” (Planta-Düngemittel GmbH) at 3 g/L and Terraflor®-AZ (Terraflor GmbH) at 0.4 g/L were added to the water up to three times a week, depending on the development of the plants.

ACCLIMATION PERIOD:

EFFECT PARAMETERS MEASURED:
Fresh Weight:
The fresh weight of the above ground part of all survived plants of a pot (each pot is considered as a replicate) was determined 14 or 21 days after 50% seedling emergence in the control. If all plants of a pot were dead the value for this replicate was assessed as “0”.

Germination:
Germination was checked daily on weekdays (except weekends) until 50% of the control plants had emerged. Further checks were done weekly.

Mortality:
The number of living and dead plants was recorded 7 and 14 days or 7, 14 and 21 days after 50% seedling emergence in the control. A plant was considered dead if no living tissue could be found on the leaves or shoots. All other plants were considered living.

Phytotoxicity:
Visual phytotoxicity (e.g. chlorosis, necrosis, abnormal growth) was recorded weekly according to EPPO Standard PP 1/135 (3) after 50% seedling emergence in the control.

Growth Stages:
Growth stages at day 14 or 21 after 50% seedling emergence in the control were reported according to BBCH-Monograph - Growth stages.

- Phytotoxicity rating system: EPPO Standard PP 1/135 (3)

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3
- Range finding study: yes
- Test concentrations in the definitive study: Based on the results of the non-GLP range finder the following concentrations of 400112 were prepared:
1,000 / 333 / 111 / 37.0 / 12.3 / 4.12 / 1.37 mg test item/kg soil dry weight (nominal)

Nominal and measured concentrations:

nominal: 1,000 / 333 / 111 / 37.0 / 12.3 mg test item/kg soil dry weight. For Allium cepa also 4.12 and 1.37 mg test item/kg soil dry weight were tested.

Reference substance (positive control):

no

Key result

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

37 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

111 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

EC10

Effect conc.:

60.3 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

179 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

1 434 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Remarks:

the EC value is extrapolated

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Brassica napus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

111 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Brassica napus

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

333 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Brassica napus

Duration:

14 d

Dose descriptor:

EC10

Effect conc.:

278 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Brassica napus

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

376 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Brassica napus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

671 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Glycine max (G. soja)

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

333 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Glycine max (G. soja)

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Helianthus annuus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

111 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Helianthus annuus

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

333 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Helianthus annuus

Duration:

14 d

Dose descriptor:

EC10

Effect conc.:

215 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Helianthus annuus

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

360 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

Helianthus annuus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

966 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

other: Solanum lycopersicon

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

111 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

other: Solanum lycopersicon

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

333 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

other: Solanum lycopersicon

Duration:

14 d

Dose descriptor:

EC10

Effect conc.:

132 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

other: Solanum lycopersicon

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

187 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Species:

other: Solanum lycopersicon

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

362 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

growth

Details on results:

Validity of the Study
Germination Rate of the Control Seeds: 83 -100%, validity criterion was met.
Mean Survival of Emerged Control Seedlings: 96 - 100%, validity criterion was met.

Growth and Morphology of the Control Seedlings:
The control seedlings exhibited no visible phytotoxic effects and the plants exhibited only normal variation in growth and morphology for that particular species; validity criterion was met. (On account of the mortality in the control for Allium cepa (4.0 %) the phytotoxicity values for these replicates are not “0”, but no further phytotoxicity was observed.)

Biological Observations
1. Brassica napus
Fresh Weight (14 days):
As control and solvent control were not statistically significant different, the fresh weight results were compared to the pooled control. The fresh weight was reduced at 37.0 mg test item/kg dry soil and at higher concentrations (-1.0% to -72.0%). This reduction was statistically significant at 333 and 1,000 mg test item/kg dry soil (-15.3% and -72.0%).
The plant development was delayed at 1,000 mg test item/kg dry soil (growth stages at test end: BBCH 13-14 in the control and BBCH 12 to 13 at 1,000 mg test item/kg dry soil).
Germination:
The germination rate ranged between 83 and 100% (control: 100%, solvent control: 97%). There was no statistically significant difference to the pooled control.
Mortality (14 days): No mortality occurred.
Phytotoxicity (14 days): Phytotoxic effects were growth reduction and leaf deformations. Strong effects were observed at 1,000 mg/kg dry soil (55%).

2. Glycine max
Fresh Weight (14 days):
As control and solvent control were statistically significant different, the fresh weight results were compared to the solvent control. The fresh weight was reduced at all tested concentrations (-0.4% to -21.5%). The reduction was statistically significant compared the solvent control at 1,000 mg test item/kg dry soil (-21.5%).
The plant development was not delayed (growth stages at test end: BBCH 12-13 in the control and BBCH 12-13 at 1,000 mg test item/kg dry soil).
Germination:
The germination rate ranged between 97 and 100% (control: 97%, solvent control: 100%). There was no statistically significant difference compared to the pooled control.
Mortality (14 days): No mortality occurred.
Phytotoxicity (14 days): The only phytotoxic effect was growth reduction and one minor leaf deformation and one minor stem deformation. The effects remained below 5% for all treatment groups.

3. Helianthus annuus
Fresh Weight (14 days):
As control and solvent control were not statistically significant different, the fresh weight results were compared to the pooled control. The fresh weight was reduced at all tested concentrations (-0.9% to -52.5%). This reduction was statistically significant at 333 and 1,000 mg test item/kg dry soil (-14.3 and -52.5%).
The plant development at the highest concentrations was slightly delayed compared to the control (growth stages at test end: BBCH 14 in the control and BBCH 12-14 at 1,000 mg test item/kg dry soil.
Germination:
The germination rate ranged between 90 and 97% (controls: 93%). There was no statistically significant difference to the pooled control.
Mortality (14 days): No mortality occurred.
Phytotoxicity (14 days):The only phytotoxic effect was growth reduction and chlorosis (and in one pot leaf deformation). Effects were observed at 1,000 mg test item/kg dry soil (15%).

4. Solanum lycopersicum
Fresh Weight (14 days):
As control and solvent control were not statistically significant different, the fresh weight results were compared to the pooled control. The fresh weight was reduced at 37.0 mg test item/kg dry soil and at higher concentrations (-6.2 to -90.7%). This reduction was statistically significant at 333 and 1,000 mg test item/kg dry soil (-45.5 and -90.7%, respectively).
The plant development was not delayed compared to the control (growth stages at test end: BBCH 12-13 in the control and BBCH 12 at 1,000 mg test item/kg dry soil; the solvent control showed a reduced BBCH 10-12, this effect did not extend to the fresh weight).
Germination:
The germination rate ranged between 73 and 100% (control: 87%, solvent control: 97%). There was no statistically significant difference compared to the pooled control.
Mortality (14 days): Mortality was observed at 111 mg test item/kg dry soil (3%) and at 1,000 mg test item/kg dry soil (14%). The effects were not statistically significant compared to the control.
Phytotoxicity (14 days): The main phytotoxic effect was growth reduction, in a few pots necrosis occured. Clear effects were observed at 333 and at 1,000 mg test item/kg dry soil (13% and 83%, respectively).

5. Avena sativa
Fresh Weight (14 days):
As control and solvent control were not statistically significant different, the fresh weight results were compared to the pooled control. The fresh weight was reduced at all tested concentrations (-3.9 to -44.8%). The reduction was statistically significant at 111 mg test item/kg dry soil and at higher concentrations (-15.6 to -44.8%).
The plant development was not delayed compared to the control (growth stages at test end: BBCH 12-13 in the controls and BBCH 12 at 1,000 mg test item/kg dry soil).
Germination:
The germination rate ranged between 93 and 100% (control: 100%, solvent control: 97%). There was no statistically significant difference compared to the pooled control.
Mortality (14 days): No mortality occurred.
Phytotoxicity (14 days):
The only phytotoxic effect was growth reduction. The maximum effect was 13% at 1,000 mg test item/kg dry soil.

6. Allium cepa
Fresh Weight (21 days):
As control and solvent control were not statistically significant different, the fresh weight results were compared to the pooled control. The fresh weight was reduced at 333 and at 1,000 mg test item/kg dry soil (-1.0% and -21.4%, respectively). This reduction was not statistically significant.
The plant development was not delayed compared to the control (growth stages at test end: BBCH 11-13 in the control and BBCH 12 at 1,000 mg test item/kg dry soil).
Germination:
The germination rate ranged between 90 and 97% (control: 83%; solvent control: 100%). There was no statistically significant difference to the pooled control.
Mortality (21 days): Mortality was observed at 333 mg test item/kg dry soil (3%) and at 1,000 mg test item/kg dry soil (4%). The effects were not statistically significant compared to the control.
Mortality was also observed at the control and solvent control (4% and 3%, respectively).
Phytotoxicity (21 days):
Phytotoxic effects at 333 mg test item/kg dry soil (8%) and 1,000 mg test item/kg dry soil (17%) were chlorosis, growth reduction and necrosis. The phytotoxic effects in the controls were due to mortality in the control. No further phytotoxicity was observed in the control.

Validity criteria fulfilled:

yes

Conclusions:

400112 was tested for effects on seedling emergence and seedling growth of six plant species out of six different plant families.
The most sensitive species in terms of fresh weight were Solanum lycopersicum, Brassica napus and Helianthus annuus with EC50 values of 362, 671 and 966 mg test item/kg dry soil, respectively (EC20 values of 187, 376 and 360 mg test item/kg dry soil, respectively). For Avena sativa the EC50 value was extrapolated to 1434 mg test item/kg dry soil. For Glycine max and Allium cepa the effects were too low to calculate effect concentration values.
The resulting NOEC (and LOEC) values were 37 (111) mg test item/kg dry soil for Avena sativa, 111 (333) mg test item/kg dry soil for Brassica napus, Helianthus annuus, and Solanum lycopersicon, 333 (1000) mg test item/kg dry soil for Glycine max, and ≥ 1000 (>1000) mg test item/kg dry soil.

Executive summary:

400112 was tested for effects on seedling emergence and seedling growth of six plant species out of six different plant families. The test was conducted according to OECD guideline 208 under GLP. The following species were tested: Solanum lycopersicum, Brassica napus, Helianthus annuus, Glycine max, Allium cepa, and Avena sativa. Based on a non GLP range finding test the following concentrations of test item 400112 were tested: 1,000 / 333 / 111 / 37.0 / 12.3 mg test item/kg soil dry weight. For Allium cepa also 4.12 and 1.37 mg test item/kg soil dry weight were tested. As 400112 is not water soluble acetone was used as carrier. The test item was mixed into the soil before sowing. 30 seeds were tested per concentration and species. The exposure time was 14 to 21 days after 50% germination in the control depending on the growth of the seedlings. The endpoints were: growth (fresh weight), germination, mortality, and phytotoxicity. The most sensitive species in terms of fresh weight were Solanum lycopersicum, Brassica napus and Helianthus annuus with EC₅₀ values of 362, 671 and 966 mg test item/kg dry soil, respectively (EC₂₀ values of 187, 376 and 360 mg test item/kg dry soil, respectively). For Avena sativa the EC₅₀ value was extrapolated to 1434 mg test item/kg dry soil. For Glycine max and Allium cepa the effects were too low to calculate effect concentration values. The resulting NOEC (and LOEC) values were 37 (111) mg test item/kg dry soil for Avena sativa, 111 (333) mg test item/kg dry soil for Brassica napus, Helianthus annuus, and Solanum lycopersicon, 333 (1000) mg test item/kg dry soil for Glycine max, and ≥ 1000 (>1000) mg test item/kg dry soil.The germination rate and the mortality were not statistically significantly reduced for any species tested. The main phytotoxic symptom was growth reduction. Necrosis, chlorosis and deformations of leaves or stems were observed occasionally or seldom.

The validity criteria were fulfilled.

Description of key information

400112 was tested for effects on seedling emergence and seedling growth of six plant species out of six different plant families. The test was conducted according to OECD guideline 208 under GLP. The following species were tested: Solanum lycopersicum, Brassica napus, Helianthus annuus, Glycine max, Allium cepa, and Avena sativa. Based on a non GLP range finding test the following concentrations of test item 400112 were tested: 1,000 / 333 / 111 / 37.0 / 12.3 mg test item/kg soil dry weight. For Allium cepa also 4.12 and 1.37 mg test item/kg soil dry weight were tested. As 400112 is not water soluble acetone was used as carrier. The test item was mixed into the soil before sowing. 30 seeds were tested per concentration and species. The exposure time was 14 to 21 days after 50% germination in the control depending on the growth of the seedlings. The endpoints were: growth (fresh weight), germination, mortality, and phytotoxicity. The most sensitive species in terms of fresh weight were Solanum lycopersicum, Brassica napus and Helianthus annuus with EC50 values of 362, 671 and 966 mg test item/kg dry soil, respectively (EC20 values of 187, 376 and 360 mg test item/kg dry soil, respectively). For Avena sativa the EC50 value was extrapolated to 1434 mg test item/kg dry soil. For Glycine max and Allium cepa the effects were too low to calculate EC (effect concentration) values. The resulting NOEC (and LOEC) values were 37 (111) mg test item/kg dry soil for Avena sativa, 111 (333) mg test item/kg dry soil for Brassica napus, Helianthus annuus, and Solanum lycopersicon, 333 (1000) mg test item/kg dry soil for Glycine max, and ≥ 1000 (>1000) mg test item/kg dry soil.

The germination rate and the mortality were not statistically significantly reduced for any species tested. The main phytotoxic symptom was growth reduction. Necrosis, chlorosis and deformations of leaves or stems were observed occasionally or seldom.

The validity criteria were fulfilled.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for terrestrial plants:

37 mg/kg soil dw