Linseed oil, oxidized

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 June - 1 October 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to established guideline and under GLP. Validity criteria were met, deviations from protocol did not adversely affect study integrity.

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not required

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil: Defined quantities of the test item (1 to 20 g nominal) was weighed into 50 mL volumetric flasks, made up with acetone and stirred for 5 minutes to completely dissolve the test item. Volumes of 40 mL of the test solutions were used to spike portions of quartz sand for the respective treatment (162 g sand per 16.2 kg soil dw). After complete evaporation of the solvent, the spiked quartz sand was mixed into the bulk test soil for the respective treatment and mixed thoroughly with an electric hand mixer.
Controls: 1: untreated soil; 2: solvent control was mixed with the same amount of quartz sand spiked with acetone but without test item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 40 ml of the solutions in acetone were mixed with 162 g sand to end up in 16.2 kg of soil (dw). The solvent is evaporated completely before mixing of the coated quartz sand through the artificial soil.
- Evaporation of vehicle before use: yes

Species:

Allium cepa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Onion
- Plant family: Amaryllidaceae
- Variety: Stuttgarter Riesen
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim Germany
- Prior seed treatment/sterilization: none

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oat
- Plant family: Poaceae
- Variety: Champion
- Source of seed: Saaten Union GmbH &, Hannover, Germany
- Prior seed treatment/sterilization: none

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Oilseed rape
- Plant family: Brassicaceae
- Variety: Liforum
- Source of seed: Landwirtschaftliche Untersuchungs- und Forschungsanstalt (LUFA), Speyer, Germany
- Prior seed treatment/sterilization: none

Species:

Cucumis sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Cucumber
- Plant family: Cucurbitaceae
- Variety: Delikatess
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim Germany
- Prior seed treatment/sterilization: none

Species:

Pisum sativum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Pea
- Plant family: Fabaceae
- Variety: Rondo
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim Germany
- Prior seed treatment/sterilization: none

Species:

other: Solanum lycopersicum

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Tomato
- Plant family: Solanaceae
- Variety: Hellfrucht
- Source of seed: Enza Zaden Deutschland GmbH & Co. KG, Dannstadt-Schauernheim Germany
- Prior seed treatment/sterilization: none

Test type:

other: Seedling emergence and growth

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

21 d

Remarks:

Starting point of test was the day where at least 50% of the seedlings had emerged in the untreated control of the species (day 0). This was 2 to 6 days after sowing, depending on the species. Final observations at 14 or 15 days after day 0.

Test temperature:

- Day/night temperatures: mean 23 (21-26) °C for B. napus; for other species: 27 (23-32) °C // acc. to study plan 22 ± 10°C

pH:

pH (0.01 M CaCl2) 5.7 ± 0.6

Moisture:

'sufficiently moistened'

Details on test conditions:

TEST SYSTEM
- Testing facility: ECT Oekotoxikologie GmbH, Böttgerstr. 2 - 14, 65439 Flörsheim am Main, Germany
- Test container (type, material, size): Common plant pots made of polypropylene (diameter 11 cm, height 8.5 cm). Each pot was placed in a separate polystyrene beaker (Kastelplast GmbH, Mainz, Germany) serving as water reservoir.
- Amount of soil: 480 ± 10 g fresh soil
- 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: 24 for Monocotyledons and 21 for Dicotyledons
- No. of replicates per treatment group: at least 4 pots with up to 6 seeds per pot. Total number of seeds per species/treatment was at least 20.
- No. of replicates per control: at least 4 pots with up to 6 seeds per pot. Total number of seeds per species/treatment was at least 20.
- No. of replicates per vehicle control: at least 4 pots with up to 6 seeds per pot. Total number of seeds per species/treatment was at least 20.

SOURCE AND PROPERTIES OF SOIL
- Geographic location: uncultivated field at Offenbach, Rhineland-Palatine, Germany
- Supplied by: Landwirtschaftliche Untersuchungs- und Forschungsanstalt (LUFA), D-67346 Speyer, Germany
- Standard soil LUFA Type 2.3, a sandy loam
- Pesticide use history at the collection site: unclutivated and no pesticides applied during last four years
- Fertiliser: in 2014, 3500 kg CaO/ha was applied three times, in June, September, December and 1463 kg MgO/ha in December
- Collection procedures: collected and sieved (=<2 mm).
- Soil texture (if natural soil): sandy loam
- Particle size distribution: < 0.002 mm: 6.5 ± 1.6 %; 0.002 - 0.05 mm: 33.8 ± 0.1 %; 0.05 - 2.0 mm: 59.7 ± 1.6 %
- Soil taxonomic classification: LUFA SOil Type 2.3; Sandy loam
- Soil classification system: USDA
- Organic carbon (%): 0.67 ± 0.03 %
- Maximum water holding capacity (in % dry weigth): 35.4 ± 1.5
- CEC: 7.5 ± 0.9 mval/100 g
- Pretreatment of soil: sieved (=< 2 mm)
- Storage (condition, duration): ambient temperature, between 10 March 2015 and 29 June 2015

Nutrient solution
- Dilution of Substral (Scotts Celaflor GmbH, Mainz, Germany), 6 ml/L water

GROWTH CONDITIONS
- Photoperiod: 16 h
- Light source: SON-T-Agro high pressure metal halide, Pilips GmbH, Hamburg Germany
- Light intensity and quality: For B. napus: 206-275 microE per m2 per sec; For other species: 207 - 360 microE per m2 per sec // acc. to study plan: => 200 microE per m2 per sec
- Day/night temperatures: mean 23 (21-26) °C for B. napus; for other species: 27 (23-32) °C // acc. to study plan 22 ± 10°C
- Relative humidity (%): 53 (35-83) % for B. napus; for other species 38 (26-67)% // acc. to study plan 70% ± 25%

- Watering regime and schedules:
- Water source/type: After placing the pots in the test room, the water reservoirs were filled with deionised water to moisten the soil sufficiently. During the test, deionised water was added to the reservoirs as necessary to maintain the soil sufficiently moist. Seedlings were fertilised once during the test by adding volume of 25 mL, of a nutrient solution to the reservoirs.
The nutrient solution was prepared by diluting a commercially available liquid fertilizer, Substral® (Scotts Celaflor GmbH, Mainz, Germany) with deionised water (6.0 mL/L water).

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

- Phytotoxicity rating system/ Scoring scheme for the visual evaluation of the test species.
Category, Evaluation Description
1 Healthy Plant with normal growth, without damages apart from what is within the normal range of variability (e.g. wilted cotyledons are no damage).
2 Slightly damaged Chlorotic and/or necrotic leaves (less than ¼ of the plant).
3 Slightly damaged Single wilted leaves (less than ¼ of the plant)
4 Severly damaged Chlorotic and/or necrotic and/or wilted leaves (more than ¼ of the plant).
5 Severely damaged Plant with developmental abnormalities.
6 Dead All above-ground parts of the plant are wilted.


VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5 or 3
- Test concentrations: 49.4 - 148 - 444 - 667 - 1000 mg/kg soil dw
Range finding study
- Test concentrations: 1 - 10 - 50 - 100 - 500 - 1000mg/kg soil ds
- Results used to determine the conditions for the definitive study: Effects were observed starting at 500 mg/kg soil for A. sativa, B. napus, C. sativus and S. esculentum. For A. cepa effects were observed at 1000 mg/kg, whereas for P. sativum no effects were observed up to 1000 mg/kg

Nominal and measured concentrations:

Nominal and measured concentrations (by weight) differed less than 2%

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

Allium cepa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

EC10

Effect conc.:

398 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Remarks on result:

other: 95% CL 205 - 512 mg/kg dw

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

148 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Species:

Avena sativa

Duration:

18 d

Dose descriptor:

LOEC

Effect conc.:

444 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

EC10

Effect conc.:

468 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Remarks on result:

other: 95% CL 279-576 mg/kg dw

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

NOEC

Effect conc.:

444 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

NOEC

Effect conc.:

444 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Key result

Species:

Brassica napus

Duration:

17 d

Dose descriptor:

EC10

Effect conc.:

335 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Remarks on result:

other: 95% CL 84-432 mg/kg dw

Species:

Cucumis sativus

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

Cucumis sativus

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

Cucumis sativus

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

667 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

Cucumis sativus

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

444 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Species:

Pisum sativum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

Pisum sativum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

Pisum sativum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

Pisum sativum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Species:

other: Solanum lycopersicum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

seedling emergence

Species:

other: Solanum lycopersicum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

other: survival

Species:

other: Solanum lycopersicum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot length

Species:

other: Solanum lycopersicum

Duration:

22 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

shoot fresh weight

Details on results:

SEED GERMINATION
- Percent seed germination: see window with 'Any other information'
- Root length: not measured
- Root discolouration/malformation: not reported
- Other effects: not reported

SEEDLING EMERGENCE
- Percent seedling emergence: Seedling emergence in test item treated soil on day 7 was at least 79% and did not differ significantly from seedling emergence in the solvent control (at least 81%).
- Percent survival: Survival of the seedlings was 100% for the control, solvent control and all test item treatments.
- Plant height: see window with 'Any other information'
- Dry weight: see window with 'Any other information'
- Seed development or appearance of seedlings: Seedlings were evaluated visually on day 7 and at the end of the test.
No visible damages were observed with the plants other than what can be considered to be within the range of normal variation of growth and morphology of the plant. This applies to all specimens of all six test species either growing in the control soil, solvent control soil or in any of the test item treated soils at test item concentrations up to and including 1000 mg/kg soil dry mass.

VEGETATIVE VIGOUR
- Percent seed germination: Seedling emergence in the control soil was at least 83% and survival of seedlings in the controls was 100% for all six test species. Hence, the requirements defined by the test guideline regarding emergence and survival of seedlings were fulfilled.
- Plant height: see window with 'Any other information'
- Dry weight: see window with 'Any other information'
- Abnormal seed development or appearance: Seedlings were evaluated visually on day 7 and at the end of the test.
No visible damages were observed with the plants other than what can be considered to be within the range of normal variation of growth and morphology of the plant. This applies to all specimens of all six test species either growing in the control soil, solvent control soil or in any of the test item treated soils at test item concentrations up to and including 1000 mg/kg soil dry mass.

Reported statistics and error estimates:

Probit Analysis was used to determine effective concentrations EC10, EC20 and EC50. Emergence and survival data were checked by the Fisher's exact binominal test with Bonferroni correction to determine the NOEC and LOEC. Significance level was alpha = 0.05, one sided greater .
Normal distribution was checked by Shapiro-Wilk’s Test, and homogeneity of variances by Levene’s Test. NOEC and LOEC for shoot length and shoot growth were determined applying the Williams Test for homogeneous variances, and by Welch-t Test for inhomogeneous variances. Significance level was alpha = 0.05, two-sided.
Statistical evaluations were performed using the ToxRat software Version 2.10 (ToxRat Solutions GmbH, 2010).

Statistics for EC10, EC20 and EC50 could be performed on the results for A. sativa and . napus (mg/kg soil dw):
A. sativa: Shoot fresh weight: EC10: 398 <205-512> ; EC20: 657 <509-771> ; EC50 n.d.
B. napus: Shoot length: EC10: 468 <279-576> ; EC20: 736 <609-852>; EC50: n.d.
B. napus: Shoot fresh weight: EC10: 335 <184-434> ; EC20: 448 <302-536>; EC50: 778 <681-927>

Tables with other statistics are reported under 'Any other information'

Number of seeds introduced at starting point of test, number of emerged seeds, and number of survived seedlings in the untreated control pots.

	A. cepa	A. sativa	B. napus	C. sativus	P. sativum	S. lycopersicum
Seeds introduced:	24	24	21	21	21	21
Day 0 [days after sowing]:	6	3	2	3	6	6
Emerged until day 7:	20	24	21	19	20	21
Live seedlings at test end:	20	24	21	19	20	21
Emerged [% of seeds sown]:	83	100	100	91	95	100
Survived [% of emerged]:	100	100	100	100	100	100

Seedling emergence and survival.

Test item concentration	A. cepa	A. sativa	B. napus	C. sativus	P. sativum	S. lycopersicum
[mg/kg soil]
	Number of seeds sown per treatment
	24	24	21	21	21	21
	Number of emerged seeds per treatment on day 7
Control	20	24	21	19	20	21
Solvent Control	23	22	21	17	20	20
49.4	20	24	20	19	21	19
148	22	23	21	19	21	19
444	19	23	19	20	21	21
667	19	24	20	19	21	20
1000	19	24	20	20	19	16^(a)
	Number of live seedlings at test end
Control	20	24	21	19	20	21
Solvent Control	23	22	21	17	20	20
49.4	20	24	20	19	21	19
148	22	23	21	19	21	19
444	19	23	19	20	21	21
667	19	24	20	19	21	20
1000	19	24	20	20	19	16^(a)
	Relative number of emerged seeds [% of seeds sown]
Control	83	100	100	91	95	100
Solvent Control	96	92	100	81	95	95
49.4	83	100	95	91	100	91
148	92	96	100	91	100	91
444	79	96	91	95	100	100
667	79	100	95	91	100	95
1000	79	100	95	95	91	89
	Relative number of live seedlings at test end [% of emerged]
Control	100	100	100	100	100	100
Solvent Control	100	100	100	100	100	100
49.4	100	100	100	100	100	100
148	100	100	100	100	100	100
444	100	100	100	100	100	100
667	100	100	100	100	100	100
1000	100	100	100	100	100	100

(^a) only 6 pots (18 seeds) instead of 7 pots (21 seeds) were prepared in treatment T5 (1000 mg/kg soil).

Nominal test item concentrations [mg/kg soil] and shoot length [mm]at the end of the test (mean±standard deviation)

Test item [mg/kg soil]	A.cepa	A.sativa	B.napus
Control	101 ± 17	332 ± 15	115 ± 10
Solvent Control	115 ± 29	319 ± 27	120 ± 8
49.4	105 ± 19	313 ± 15	126 ± 10
148	104 ± 13	307 ± 15	119 ± 6
444	121 ± 21	316 ± 12	111 ± 8
667	123 ± 13	313 ± 6	95.6 ± 10*
1000	121± 13	296± 13	85.7± 7*
	C. sativus	P. sativum	S. lycopersicum
Control	109± 5	151± 21	188± 7
Solvent Control	104± 4	155± 13	187± 11
49.4	109± 8	161± 11	199± 14
148	110± 5	155± 9	194± 8
444	103± 6	165± 13	190± 11
667	97.0± 4*	155± 19	187± 11
1000	89.3± 3*	160± 16	185± 13

Nominal test item concentrations [mg/kg soil] and shoot fresh weight [g] at the end of the test (mean and standard deviation).

Test item [mg/kg soil]	A. cepa	A. sativa	B. napus
Control	0.135± 0.038	0.748± 0.037	1.46± 0.158
Solvent Control	0.165± 0.070	0.757± 0.048	1.37± 0.094
49.4	0.128± 0.039	0.744± 0.065	1.51± 0.330
148	0.142± 0.024	0.730± 0.027	1.39± 0.173
444	0.170± 0.034	0.662± 0.041*	1.15± 0.231
667	0.166± 0.029	0.621± 0.037*	0.742± 0.138*
1000	0.165± 0.021	0.506± 0.021*	0.517± 0.079*
	C. sativus	P. sativum	S. lycopersicum
C	2.14± 0.20	1.51± 0.29	2.34± 0.26
SC	2.15± 0.28	1.58± 0.16	2.34± 0.35
49.4	2.17± 0.33	1.68± 0.09	2.78± 0.70
148	2.51± 0.29	1.61± 0.13	2.65± 0.37
444	2.11± 0.19	1.68± 0.11	2.49± 0.22
667	1.80± 0.25*	1.54± 0.20	2.47± 0.24
1000	1.49± 0.18*	1.66± 0.22	2.16± 0.53

(*) = significantly different from the solvent control (SC) (p <0.05, two sided).

Validity criteria fulfilled:

yes

Remarks:

1. More than 70% of the controls emerged. 23 Less than 10% of the control plants died. 3. No phytotoxic effects were observed in the controls. 4.Environmental conditions per species were identical and growing media contained the same amount of soil

Conclusions:

For all species the NOEC for seedling emergence and survival was at or above 1000 mg/kg soil dw.
Out of the 6 test species, shoot length was affected in two test species and shoot fresh weight was affected in three test species, with the lowest NOEC and LOEC of 148 and 444 mg/kg soil dw, respectively (shoot fresh weight of Avena sativa, oat). The accompanying EC10 was 398 mg/kg dw <95%C.L. 205-512 mg/kg dw>.
The lowest EC10 of 335 mg/kg dw <95% C.L. 184-432 mg/kg dw> was observed with the shoot fresh weight in Brassica napus (rape).

Executive summary:

The toxicity of Blow Linseed Oil, oxidized, was studies on six plant species according to OECD Test Guideline 208: Terrestrial Plant Test: Seedling Emergence and Seedling Growth Test. The study was conducted in compliance with the Principles for Good Laboratory Practice

Methodology

Seeds of two monocotyledonous species, Avena sativa (oats) and Allium cepa (onion), and four dicotyledonous species, Brassica napus (oilseed rape), Cucumis sativus (cucumber), Pisum sativum (peas) and Solanum lycopersicum (tomato) were planted in a natural sandy loam soil (standard soil LUFA Sp2.3) immediately after test item application and left to grow under controlled conditions for 14 to 15 days. Each species was tested at 5 test item concentrations: 49.4, 148, 444, 667 and 1000 mg test item/kg soil dry weight (dose response test). The test item was dissolved in acetone, spiked onto quartz sand and mixed thoroughly into the soil. The control was untreated soil, while the solvent control was mixed with the same amount of acetone spiked quartz sand without test item. Plants were allowed to emerge and grow for 14 to 15 days following 50% emergence of the control plants. Soils were supplied with water or nutrient solution by bottom watering. The test was performed in growth chambers equipped with artificial lighting for 16 hours per day and air conditioning. The pots were placed randomly at the beginning of the test and were re-arranged at minimum weekly intervals. On day 7 after 50% emergence, seedlings were evaluated visually. At test end, final counting, visual evaluation, and harvest to determine shoot length and shoot fresh weight took place.

The validity criteria were fulfilled as seedling emergence in the control was ≥ 70% and seedling survival in the control was ≥ 90% for all six species. No phytotoxicity was observed in the controls.

Emergence and Survival

Seedling emergence in the control soil was at least 83%. Seedling emergence in test item treated soil was at least 79% and did not differ significantly from seedling emergence in the solvent control (at least 81%).

Survival of the seedlings was 100% for the control, solvent control and all test item treatments.

Visual appearance

No damage was observed other than what can be considered to be within the range ofnormal variation of growth and morphology of the plant.

Shoot length and fresh weight

Shoot length was significantly reduced at test item concentrations of ≥667 mg/kg soil in B. napus and C. sativus. No effects on shoot length were observed in A. cepa, A. sativa, P. sativum and S. lycopersicum at concentrations up to and including 1000 mg/kg soil.

Shoot fresh weight was significantly reduced at ≥444 mg/kg soil in A. sativa and at ≥667 mg/kg soil in B. napus and C. sativus. No effects on shoot fresh weight were observed in A. cepa, P. sativum and S. lycopersicum at concentrations up to and including 1000 mg/kg soil.

Conclusion:      

The No-Observed-Effect-Concentration (NOEC) with regard to seedling emergence and survival of seedlings was at or above 1000 mg/kg soil dw.

With regard to shoot length two test species and with regard to shoot fresh weight three out of the six test species were affected, resulting in a lowest NOEC and LOEC of 148 and 444 mg/kg soil dry mass, respectively, observed with the fresh weight of Avena sativa (oat). The accompanying EC₁₀ was 398 mg/kg dw <95%C.L. 205-512 mg/kg dw>.

The lowest EC₁₀of 335 mg/kg dw <95%C.L. 184 - 432 mg/kg dw> was observed with the shoot fresh weight in Brassica napus (rape).

Description of key information

Out of the 6 test species, shoot length was affected in two test species and shoot fresh weight was affected in three test species, with the lowest NOEC and LOEC of 148 and 444 mg/kg soil dw, respectively. The accompanying EC10 was 398 mg/kg dw <95% C.L. 205-512 mg/kg dw>.

The lowest EC10 of 335 mg/kg <95% C.L. 184-432 mg/kg dw> was observed with the shoot fresh weight in another species.

Key value for chemical safety assessment

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

335 mg/kg soil dw

Additional information

A toxicity study on six plant species was performed according to OECD TG208 and in compliance with GLP.

The emergence and growth of six test species (two monocotyledons, Avena sativa (oats) and Allium cepa (onion)) and four dicotyledons (Brassica napus (oilseed rape), Cucumis sativus (cucumber), Pisum sativum (peas) and Solanum lycopersicum (tomato)) in a natural sandy loam soil was followed for 17 to 21 days while exposed to 49.4, 148, 444, 667 and 1000 mg test item/kg soil dry weight. Seedling emergence in test item treated soil did not differ significantly from seedling emergence in the solvent control. No visual damage was observed. Shoot length was significantly reduced at test item concentrations of ≥667 mg/kg soil in B. napus and C. sativus. No effects on shoot length were observed in A. cepa, A. sativa, P. sativum and S. lycopersicum at concentrations up to and including 1000 mg/kg soil. Shoot fresh weight was significantly reduced at ≥444 mg/kg soil in A. sativa and at ≥667 mg/kg soil in B. napus and C. sativus. No effects on shoot fresh weight were observed in A. cepa, P. sativum and S. lycopersicum at concentrations up to and including 1000 mg/kg soil.

NOEC seedling emergence and survival of seedlings was at or above 1000 mg/kg soil dw for all species.

Shoot length was affected in 2 species and fresh shoot weight in 3 species.

The lowest NOEC and LOEC were 148 and 444 mg/kg soil dry mass, respectively, observed for the fresh weight of Avena sativa(oat). The accompanying EC10 was 398 mg/kg dw <95% C.L. 205-512 mg/kg dw>.

The lowest EC₁₀of 335 <184-432) mg/kg dw was observed with the shoot fresh weight in Brassica napus (rape). Thus the values of 335 mg/kg is the lowest value (EC₁₀) for plants.