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Ecotoxicological information

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:
The study was conducted between 01 March 2016 and 06 June 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)
Version / remarks:
19 July 2006
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor; Batch no. Ei 2985
- Expiration date of the lot/batch: 01 December 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient 10 - 30”C
Analytical monitoring:
no
Vehicle:
yes
Remarks:
Water
Details on preparation and application of test substrate:

- Method of mixing into soil (if used):
For preparations A the test item was weighed into beakers, the total volume of RO water was added and the containers were placed into a heated sonic bath, maintained at 40°C, until the test substance had formed a homogeneous dispersion.

For all lower rate preparations the test item was weighed into beakers, the portion of the final volume of RO water was added and the containers were placed into a heated sonic bath, maintained at 40°C, until the test substance had formed a homogeneous dispersion. The preparations were then transferred to volumetric flasks, and the beakers were rinsed with additional RO water which was then added to the flasks until the final volume had been achieved.

The resulting preparations were dense, white dispersions in the RO water, which became visibly less dense with declining concentration.

After preparation all the beakers and flasks were left in the sonic bath until the application to the soil. The flasks were also shaken between applications.

The prepared test item was mixed into the soil using a hand held electric mixer before it was distributed among labelled replicate pots.

The treatments were applied in the order of water control then the test item at nominal concentrations of 333.33, 1.37, 4.12, 12.34, 37.04, 111.11 and 1000 mg/kg dry soil for all species. The blades of the mixer were rinsed in RO water between treatment rates.

- Controls:
For the untreated water control group the same amounts of moist soil and volumes of RO water were used as for the treatment groups.

- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Water
Species:
Brassica oleracea var. capitata
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Cabbage
- Plant family: Cruciferae (Brassicaceae)
Species:
Daucus carota
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Carrot
- Plant family: Apiaceae (Umbelliferae)
Species:
Lactuca sativa
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Lettuce
- Plant family: Asteraceae (Compositae)
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Tomato
- Plant family: Solanaceae
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: Oat
- Plant family: Poaceae (Gramineae)
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: Onion
- Plant family: Liliaceae (Amarylladaceae)
Test type:
seedling emergence and seedling growth test
Study type:
laboratory study
Substrate type:
natural soil
Remarks:
Sandy Loam
Limit test:
no
Total exposure duration:
23 d
Remarks:
Each plant species was sown into treated soil and monitored for 14 - 19 days following a minimum of 50% germination of the control seedlings.
Test temperature:
18.0 - 30.2 ”C (mean 21.0 - 24.0”C)
pH:
6.8 - 7.4
Moisture:
9.41% prior to application of test item
Details on test conditions:
TEST SYSTEM
- Testing facility:
- Test container (type, material, size): 10 cm i.d. plastic pots
- Amount of soil: filll to depth of 7.5 cm
- Method of seeding: seeds were spaced out on the soil surface and approximately 5 mm of the same soil was added to cover the seeds
- No. of seeds per container: three seeds were used for cabbage, carrot and lettuce, two seeds for tomato and four seeds for oat and onion.
- No. of plants (retained after thinning): no thinning
- No. of replicates per treatment group: 14 replicates for cabbage, carrot and lettuce, 20 replicates for tomato and 10 replicates for oat and onion

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)

- Soil texture (if natural soil)
- % sand: 79% w/w
- % silt: 10% w/w
- % clay: 11% w/w
- Soil taxonomic classification: Sandy loam
- Soil classification system:
- Organic carbon (%): 1.1% w/w


GROWTH CONDITIONS
- See Table 1 below:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

Each pot was inspected daily and the number of emerged plants recorded, together with any herbicidal symptoms. Emergence was defined as any plant material observed above the soil surface. After 50% emergence in the controls was achieved, observations continued every one to two days until the plants were harvested. Observations included visual assessments based on the overall condition of surviving seedlings. These assessments were conducted using a uniform scoring system based on plant size and vigour, where 0 indicated no abnormal effects and 10 indicated plant death (Appendix 5).

Plants from all treatment and control groups were harvested 14-19 days after germination of at least 50% of the control seedlings, by cutting each stem at soil level, giving a total exposure period of 21 days for cabbage and onion, 22 days for carrot and lettuce and 23 days for tomato and oat. The height of each plant was then recorded before they were placed into individually labelled paper bags and dried to a constant weight at approximately 60ºC at which time the dry weight was recorded. The height of dicotyledons was measured to the growing point and the length of the longest leaf was recorded for monocotyledons.


VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS

- Range finding study
- Test concentrations: A preliminary range finder phase was carried out at test concentrations of 0.1, 1, 10, 100 and 1000 mg/kg dry soil. For cabbage, carrot and lettuce five replicate pots of three seeds were sown for tomato eight pots of two seeds were sown, with four pots of four seeds for oat and onion.
- Results used to determine the conditions for the definitive study: Six treatments were applied for each species: a water control and the test item at rates of 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species. Additional treatments at 1.37 and 4.12 mg/kg dry soil were applied for lettuce, oat and onion.
Nominal and measured concentrations:
Six treatments were applied for each species: a water control and the test item at rates of 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species. Additional treatments at 1.37 and 4.12 mg/kg dry soil were applied for lettuce, oat and onion.
Reference substance (positive control):
no
Key result
Species:
Brassica oleracea var. capitata
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:
other: Height
Key result
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
111.11 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Key result
Species:
Lactuca sativa
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Key result
Species:
Lycopersicon esculentum
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Key result
Species:
Avena sativa
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Key result
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.
Basis for effect:
other: Height
Key result
Species:
Brassica oleracea var. capitata
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: dry weight
Key result
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry weight
Key result
Species:
Lactuca sativa
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry weight
Key result
Species:
Lycopersicon esculentum
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry weight
Key result
Species:
Avena sativa
Duration:
23 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry weight
Key result
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.
Basis for effect:
other: Dry weight
Details on results:
Range finding
There were no treatment related effects on the germination or survival of any species at rates up to 1000 mg/kg dry soil.

A statistically significant reduction in height was recorded for carrot at 1000 mg/kg dry soil. Statistically significant reductions in dry weight were recorded for cabbage, carrot, oat and onion at 1000 mg/kg dry soil.

The EC25 values for effects on height were >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 264.60 mg/kg dry soil for carrot. The EC50 values for height were >1000 mg/kg dry soil for all species. The EC25 values for effects on dry weight were 877.75, 823.34, 892.80, 474.19 and 16.93 mg/kg dry soil for cabbage, carrot, lettuce, tomato and onion respectively and >1000 mg/kg dry soil for oat. The EC50 values for effects on dry weight were 949.19 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species.

Effect on germination
There were no treatment-related effects on plant germination at rates of up to 1000 mg/kg dry soil for any of the species tested.

Germination in the water controls was 100% for cabbage, carrot, tomato and oat, 97.6% for lettuce and 77.5% for onion.
A statistically significant reduction in germination (16.7%) in comparison to the water control was recorded for cabbage at 333.33 mg/kg dry soil, however the 7.1% reduction in germination at 1000 mg/kg dry soil was not statistically significant. The observed effect at 333.33 mg/kg dry soil is therefore not considered to be treatment related.

Effects on plant survival
There was 100% survival of all species in the water controls and for carrot, lettuce, tomato and oat at all rates of application up to and including 1000 mg/kg dry soil. Minor effects on plant survival were recorded for cabbage (2.4 and 5.1% plant mortality at 111.11 and 1000 mg.kg dry soil respectively) and onion (3.1 and 2.8% plant mortality at 1.37 and 333.33 mg/kg dry soil respectively). These effects at rates <1000 mg/kg dry soil were not considered to be treatment related.

Effects on plant growth and health
Based on visual observations, there was no visible effect in oat at any treatment rate. Slight growth inhibition was observed in cabbage at all treatment rates, in carrot and lettuce at
1000 mg/kg dry soil, in tomato at 333.33 and 1000 mg /kg dry soil and in onion at 37.04, 333.33 and 1000 mg/kg dry soil, (all <10%).

The EC25 values for height were estimated to be >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 387.92 mg/kg dry soil for carrot. The EC50 values for height were estimated to be >1000 mg/kg dry soil for all species. The NOEC values for effects on height were 1000 mg/kg dry soil for cabbage, lettuce, oat and onion, 111.11 mg/kg dry soil for carrot and 333.33 mg/kg dry soil for tomato.

The EC25 values for dry weight were estimated to be 800.42 and 548.29 mg/kg dry soil for cabbage and carrot respectively and >1000 mg/kg dry soil for lettuce, tomato, oat and onion. The EC50 values for dry weight were estimated to be 878.35 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species. The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage and carrot and 1000 mg/kg dry soil for lettuce, tomato, oat and onion.

The study was considered valid as the validation criteria were met and no visible treatment related effects were observed in the controls.
Results with reference substance (positive control):
N/A
Validity criteria fulfilled:
yes
Conclusions:
There were no treatment-related effects on plant germination or survival for carrot, lettuce, tomato, oat or onion for all species at rates of up to 1000 mg/kg dry soil. There was a 16.7% reduction in germination for cabbage at 333.33 mg/kg dry soil and a 5.1% reduction in survival at 1000 mg/kg dry soil.

Based on visual observations, there were no effects of >10% at rates up to 1000 mg/kg dry soil for any of the species tested.
The EC25 values for height were estimated to be >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 387.92 mg/kg dry soil for carrot. The EC50 values for height were estimated to be >1000 mg/kg dry soil for all species. The NOEC values for effects on height were 1000 mg/kg dry soil for cabbage, lettuce, oat and onion, 111.11 mg/kg dry soil for carrot and 333.33 mg/kg dry soil for tomato.

The EC25 values for dry weight were estimated to be 800.42 and 548.29 mg/kg dry soil for cabbage and carrot respectively and >1000 mg/kg dry soil for lettuce, tomato, oat and onion. The EC50 values for dry weight were estimated to be 878.35 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species. The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage and carrot and 1000 mg/kg dry soil for lettuce, tomato, oat and onion.

The study was considered valid as the validation criteria were met and no visible treatment related effects were observed in the controls.
Executive summary:

This study was carried out with four dicotyledons, (cabbage, carrot, lettuce and tomato) and two monocotyledons, (oat and onion), to determine the toxicity of TOFA_TETA_PAA_BADGE_CGE_Adduct (hereafter referred to as the test item in the text and TOFA in tables) to seedling emergence and seedling growth of non-target plant species during early critical stages in their development, in a dose response test designed to evaluate the phytotoxic effects of the test substance. Each plant species was sown into treated soil and monitored for 14 - 19 days following a minimum of 50% germination in the controls.

Following initial range finding at 0.1, 1, 10, 100 and 1000 mg/kg dry soil, the definitive study was conducted as a dose response test at nominal rates of 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species. Additional rates of 1.37 and 4.12 mg/kg dry soil were included for lettuce, oat and onion.

For cabbage, carrot and lettuce 14 replicate pots of three seeds were sown for each treatment rate, with 20 pots of two seeds for tomato and ten pots of four seeds for oat and onion. An untreated water control was also included for all species. The soil used was a sandy loam, sieved to remove all particles > 2 mm.

After application and sowing, the pots were checked daily and the numbers of emerged seedlings were recorded. Emergence, survival and sub-lethal effects were recorded daily up to a minimum of 50% emergence of control seedlings and thereafter every one to three days. Plants were harvested 14 - 19 days after 50% control germination, giving a total exposure period of 21 days for cabbage and onion, 22 days for carrot and lettuce and 23 days for tomato and oat. The height of each individual plant was then measured before the plants were dried and weighed to obtain a measurement of dry weight.

Effect on germination and plant survival

Germination in the water controls was 100% for cabbage, carrot, tomato and oat; 97.6% for lettuce and 77.5% for onion. A statistically significant reduction in germination (16.7%) in comparison to the water control was recorded for cabbage at 333.33 mg/kg dry soil.

There was 100% survival of all species in the water controls and for carrot, lettuce, tomato and oat at all rates of application up to and including 1000 mg/kg dry soil. Minor effects on plant survival were recorded for cabbage (2.4 and 5.1% plant mortality at 111.11 and 1000 mg.kg dry soil respectively) and onion (3.1 and 2.8% plant mortality at 1.37 and 333.33 mg/kg dry soil respectively). Where these effects occurred at rates <1000 mg/kg dry soil they were not considered to be treatment related.

Effect on plant growth and health

At harvest, measurement of the height and dry weight for each surviving seedling from each replicate pot was conducted. Following statistical analysis significant reductions in height were estimated for carrot at 333.33 and 1000 mg/kg dry soil and for tomato at 1000 mg/kg dry soil in comparison to the water control. Significant reductions in dry weight were estimated for cabbage and carrot at 1000 mg/kg dry soil. There were no statistically significant adverse effects in either height or dry weight for lettuce, oat and onion at rates up to 1000 mg/kg dry soil.

Based on visual observations, there were no effects of >10% at rates up to 1000 mg/kg dry soil for any of the species tested. The EC25 values for effects on height were estimated to be >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 387.92 mg/kg dry soil for carrot.

The EC50 values for effects on height were estimated to be >1000 mg/kg dry soil for all species. The NOEC values for effects on height were 1000 mg/kg dry soil for cabbage, lettuce, oat and onion, 111.11 mg/kg dry soil for carrot and 333.33 mg/kg dry soil for tomato. The EC25 values for effects on dry weight were estimated to be 800.42 and 548.29 mg/kg dry soil for cabbage and carrot respectively and >1000 mg/kg dry soil for lettuce, tomato, oat and onion.

The EC50 values for the effects on dry weight were estimated to be 878.35 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species. The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage and carrot and 1000 mg/kg dry soil for lettuce, tomato, oat and onion.

Conclusion

There were no treatment-related effects on plant germination or survival for carrot, lettuce, tomato, oat, onion for all species at rates of up to 1000 mg/kg dry soil. There was a 16.7% reduction in germination for cabbage at 333.33 mg/kg dry soil and a 5.1% reduction in survival at 1000 mg/kg dry soil.

The EC25 values for effects on height were estimated to be >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 387.92 mg/kg dry soil for carrot. The EC50 values for effects on height were estimated to be >1000 mg/kg dry soil for all species. The NOEC values for effects on height were 1000 mg/kg dry soil for cabbage, lettuce, oat and onion, 111.11 mg/kg dry soil for carrot and 333.33 mg/kg dry soil for tomato.

The EC25 values for effects on dry weight were estimated to be 800.42 and 548.29 mg/kg dry soil for cabbage and carrot respectively and >1000 mg/kg dry soil for lettuce, tomato, oat and onion. The EC50 values for effects on dry weight were estimated to be 878.35 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species. The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage and carrot and 1000 mg/kg dry soil for lettuce, tomato, oat and onion.

The study was considered valid as the validation criteria were met and no visible treatment related effects were observed in the controls.

Description of key information

There were no treatment-related effects on plant germination or survival for carrot, lettuce, tomato, oat or onion for all species at rates of up to 1000 mg/kg dry soil. There was a 16.7% reduction in germination for cabbage at 333.33 mg/kg dry soil and a 5.1% reduction in survival at 1000 mg/kg dry soil.

Based on visual observations, there were no effects of >10% at rates up to 1000 mg/kg dry soil for any of the species tested.

The EC25 values for height were estimated to be >1000 mg/kg dry soil for cabbage, lettuce, tomato, oat and onion and 387.92 mg/kg dry soil for carrot. The EC50 values for height were estimated to be >1000 mg/kg dry soil for all species. The NOEC values for effects on height were 1000 mg/kg dry soil for cabbage, lettuce, oat and onion, 111.11 mg/kg dry soil for carrot and 333.33 mg/kg dry soil for tomato.

The EC25 values for dry weight were estimated to be 800.42 and 548.29 mg/kg dry soil for cabbage and carrot respectively and >1000 mg/kg dry soil for lettuce, tomato, oat and onion. The EC50 values for dry weight were estimated to be 878.35 mg/kg dry soil for carrot and >1000 mg/kg dry soil for all other species. The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage and carrot and 1000 mg/kg dry soil for lettuce, tomato, oat and onion.

The study was considered valid as the validation criteria were met and no visible treatment related effects were observed in the controls.

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for terrestrial plants:
111.11 mg/kg soil dw

Additional information