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Ecotoxicological information

Toxicity to terrestrial plants

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Reference
Endpoint:
toxicity to terrestrial plants: short-term (with study design considered suitable for long-term assessment)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 Dec 2016 - 23 Jan 2017
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:
2006
Deviations:
yes
Remarks:
see section"Any other information on material and methods incl tables"
GLP compliance:
yes (incl. QA statement)
Remarks:
Hessisches Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil: test item were weighed out for each treatment separately and ground with fine quartz sand and afterwards mixed into the test soil; concentration of sand added to the soil was 1% (i.e. 10 g/kg soil dw).
- Controls: control soil received sand which has not been treated with the test item
- Others: Application rate in detail is shown in Tables 3-5 in section "Any other information on material and methods incl tables"
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: Onion
- Plant family: Amaryllidaceae
- Cultivar: Stuttgarter Riesen
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim, Germany
- Lot number: 4333216
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
- Common name: oat
- Plant family: Poaceae
- Lot number: n.a.
- Source of seed: Nordsaat Saatzucht GmbH, Langenstein, Germany
- Cultivar: Canyon
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
Species:
Brassica napus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: rape
- Plant family: Brassicaceae
- Lot number: n.a.
- Source of seed: Feldsaaten Freudenberger, Krefeld, Germany
- Cultivar: Akela
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
Species:
Cucumis sativus
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Cucumber
- Plant family: Cucurbitaceae
- Lot number: 4291508
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim, Germany
- Cultivar: Delikatess
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
Species:
Pisum sativum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Pea
- Plant family: Fabaceae
- Lot number: 117024NU
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim, Germany
- Cultivar: Rondo
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
Species:
other: Solanum lycopersicum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
- Common name: Tomato
- Plant family: Solanaceae
- Lot number: 4317738
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim, Germany
- Cultivar: Hellfrucht
- Seed storage: under cool and dry conditions in the dark
- Pretreatment of seeds: seeds had neither been dressed with fungicides nor imbibed with water
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 air temperature ranging from 20.4 to 24.5 °C


pH:
5.9 ± 0.6 (soil, 0.01 M CaCl2)
Details on test conditions:
TEST SYSTEM
- Testing facility: ECT GmbH
- Test container: polypropylene pots (diameter 11 cm, height 8.5 cm), placed in a separate polystyrene beaker (Kastelplast GmbH, Mainz, Germany) serving as water reservoir
- Amount of soil: 468 ± 10 g soil fresh weight
- Method of seeding: seeds were randomly picked with a pair of tweezers and pressed into the soil at a uniform distance between seeds
- No. of seeds per container: 6 (for monocotyledonous) and 3 (for dicotyledonous)
- No. of replicates per treatment group: 4 (for monocotyledonous) and 7 (for dicotyledonous)
- No. of replicates per control: at least 4 (for monocotyledonous) and 7 (for dicotyledonous)

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: standard soil LUFA Type 2.3. (Batch F 2.3 4316) was collected from an uncultivated field at Offenbach, Rhineland-Palatinate, Germany on 24 Oct 2016 (information on soil properties was provided by the Landwirtschaftliche Untersuchungs- und Forschungsanstalt (LUFA) Speyer, Germany)
- Pesticide use history at the collection site: field had neither received any organic fertilisers nor plant protection substances during the last four years before sampling
Soil texture
- % sand (0.05 – 2.0 mm): 59.6 ± 1.4
- % silt (0.002 – 0.05 mm): 33.6 ± 0.5
- % clay (< 0.002 mm): 6.8 ± 1.6
- Soil taxonomic classification: sandy loam
- Soil classification system: USDA
- Nitrogen (%): 0.08 ± 0.01
- Organic carbon (%): 0.67 ± 0.03
- Maximum water holding capacity (in % dry weigth): 35.6 ± 1.4
- CEC: 7.6 ± 0.8 meq/100 g
- Pretreatment of soil: soil was sieved (≤2 mm)
- Storage (condition, duration): soil was stored at ECT GmbH at ambient temperature until usage

NUTRIENT MEDIUM (if used)
- Description: nutrient solution (“Substral®”, 6 ml/L H2O) was added several times during the trial

GROWTH CONDITIONS
- Photoperiod: 16 h
- Light source: artificial lighting (SON-T-Agro high pressure metal halide lamps, Philips GmbH, Hamburg, Germany)
- Light intensity [μE m-2 s-1]: see Table 4 in section "Any other information on material and methods incl tables"
- Day/night temperatures: see Table 4 in section "Any other information on material and methods incl tables"
- Relative humidity (%): see Table 4 in section "Any other information on material and methods incl tables"
- Watering regime and schedules: plants were consequently watered enough to avoid dehydration of the plants
- Volume applied: deionised water was added to the reservoirs as necessary to maintain the soil sufficiently moist

EFFECT PARAMETERS MEASURED: emergence rate, survival of seedlings, shoot fresh weight, and visual detrimental effects (chlorosis, necrosis, development abnormalities). Day 0 of the test was defined as the day by which at least 50% of the seedlings in the untreated control pots of a particular species had emerged. Visual detrimental effects were assessed on day 7, day 14, and at the end of the test; Shoot fresh weight was evaluated only at the end of the test

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.16
Range finding study (non-GLP)
- Test concentrations: between 1.0 and 1000 mg/kg soil dry weight
- Results used to determine the conditions for the definitive study: Based on the results of the non-GLP range-finding test the main test was designed (results are summarized in table 1 in section "Any other information on results incl tables"
Nominal and measured concentrations:
Control, 10, 31.6, 100, 316 and 1000 mg test item /kg soil dw
Reference substance (positive control):
no
Species:
Brassica napus
Duration:
19 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: all parameters
Remarks:
emergence, survival and shoot fresh weight
Species:
Pisum sativum
Duration:
20 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: all parameters
Remarks:
emergence, survival and shoot fresh weight
Species:
other: all remaining species
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: all parameters
Remarks:
emergence, survival and shoot fresh weight
Details on results:
SEEDLING EMERGENCE
- Percent seedling emergence (untreated control pot):
79.2% (A. cepa)
87.5% (A. sativa)
90.5% (B. napus)
90.5% (C. sativus)
90.5% (P. sativum)
95.2% (S.lycopersicum)

- Percent survival (untreated control pot):
100% (A.cepa)
100% (A. sativa)
100% (B. napus)
100% (C. sativus)
94.7% (P. sativum)
100% (S. lycopersicum)

- Visual appearance: Plants growing in treated soil did not differ in their visual appearance from plants growing in non-treated control soil and did not exhibit any visible test item related damages.

Reported statistics and error estimates:
Number of emerged and number of survived seedlings and shoot fresh weight at the end of the test were considered for statistical data analysis. Determined NOEC values for emergence rate, survival as well as shoot fresh weight are >1000 mg/kg dw for every tested species. EC10, EC25 and EC50 values are assumed to be higher than >1000 mg/kg dw

Seedling Emergence and Survival

Emergence in the control was at least 79.2%. Survival of seedlings in the controls was at least 94.7%. Hence, the requirements defined by the test guideline regarding emergence and survival of seedlings were fulfilled. Neither emergence nor survival of the seedlings was statistically significantly affected in any of the species at test item concentrations up to and including 1000 mg/kg soil dry weight which was the highest tested concentration.

 

Visual Appearance

Plants growing in treated soil did not differ in their visual appearance from plants growing in non-treated control soil and did not exhibit any visible test item related damages.

 

Seedling Shoot Fresh Weight

Shoot fresh weight of plants growing in test item treated soil was not statistically significantly different from shoot weight of the plants growing in untreated control soil.

 

Validity criteria fulfilled:
yes

Description of key information

NOEC ≥1000 mg/kg soil dry weight (all parameters, all test species, OECD 208)

Key value for chemical safety assessment

Additional information

One experimental study is available investigating the potential effects of the test item (CAS No. 10094-45-8) on seedling emergence and growth (Croda, 2017) of plants. The study was performed according to OECD guideline 208 under GLP conditions. For this study six species of non-target terrestrial plants (2 monocots and 4 dicots) were planted in a natural sandy loam soil (standard soil LUFA Sp 2.3) immediately after test item application. Five test item concentrations were tested with each species: 10, 31.6, 100, 316 and 1000 mg test item /kg soil dry weight and left to grow under controlled conditions for 19 days (B. napus), 20 days (P. sativum) or 21 days (remaining species) following 50% emergence of the control plants. Test item was mixed with fine quartz sand and added to soil. The concentration of sand added to the soil was 1% (i.e. 10 g/kg soil). Soils were supplied with water and a nutrient solution as needed by bottom watering. The test was performed in growth chambers equipped with artificial lighting (201 to 352 μE m-2 s-1, for 16 hours per day) and air conditioning (air temperature ranging from 20.9 to 24.5 °C, relative humidity ranging from 22.5% to 43.1%). On day 7 and 14 or 15 (Brassica napus) after 50% emergence of control, seedlings were evaluated visually. At test end, seedlings were counted, evaluated visually, and harvest to determine shoot fresh weight. The effects of the test item on the emergence and growth of seedlings were tested in the laboratory. At concentrations up to and including 1000 mg/kg soil dry weight the test item had no statistically significant adverse effect on either seedling emergence, seedling survival or shoot fresh weight of any of the six test species. The NOEC and LOEC were therefore determined to be ≥1000 mg/kg soil dry weight and > 1000 mg/kg soil dry weight, respectively. Due to the lack of adverse effects effective concentrations could not be calculated and are assumed to be higher than 1000 mg/kg dry weight.