Linseed oil, oxidized

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 May 2015 - 27 July 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to international guideline (OECD guideline 222) under GLP. Validity criteria met, deviations from protocol did not adversely affect study integrity.

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)

Deviations:

no

Principles of method if other than guideline:

not relevant

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: The test item is poorly soluble in water; therefore, a stock solution was prepared in acetone.The concentration was based on the product Linseed oil, Oxidised. An appropriate volume of this stock emulsion was used to soak a portion of the quartz sand of the artificial soil. After evaporation of the solvent for 2 hours by placing under a fume hood, the quartz sand was coated by the test item. The test item coated fraction of the quartz sand was then incorporated into the remainder of the artificial soil by thoroughly mixing it in for approx. 10 minutes.
- Controls: 8 solvent replicates were tested
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution): The test item is coated to the quartz sand and then incorporated into the remainder of the artificial soil by thoroughly mixing it in for approx. 10 minutes. The solvent used for the coating process is assumed to be evaporated completely before mixing of the coated quartz sand through the artificial soil.
- Evaporation of vehicle before use: yes (2 hours under a fume hood)

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida (Lumbricidae, Earthworms)
- Source: ECT Oekotoxikologie GmbH since 1994
- Age at test initiation: 10 to 11 months
- Weight at test initiation: 250 to 600 mg

ACCLIMATION
- Acclimation period: 24 hours
- Acclimation conditions: Same as test
- Health during acclimation: No mortality observed
- Feeding: adult earthworms were fed weekly the first 4 weeks with 5g finely ground cow manure (Jesus Bruderschaft e.V. Gnadenthal, D-65597 Hünfelden) moistened with 10 mL deionised water was spread on the soil surface of each test container. If food remained uneaten the ration was reduced on demand. After removing the adults on day 28 a further 5 g of food moistened with 10 mL deionised water was mixed with the artificial soil of each test vessel.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

56 d

Remarks:

The adult earthworms were removed from the substrate after 28 days.

Post exposure observation period:

Not applicable.

Test temperature:

17.7 – 21.2 °C
Mean 19.3 °C

pH:

Soil pH-value (day 0): 6.2 – 6.3
Soil pH-value (day 56): 6.5

Moisture:

Soil moisture (day 0): 45.8 – 50.9% of WHCmax
Soil moisture (day 56): 53.3 – 63.2% of WHCmax
WHC = water holding capacity

Details on test conditions:

TEST SYSTEM
- Test container: Bellaplast containers (Company Kastelplast, D-55120 Mainz) consisting of inert (non-toxic) plastic (Polystyrol). Base area: 11 x 15.5 cm (= 170.5 cm²), height: 6 cm. The test vessels were covered with a transparent and perforated lid.
- Amount of soil or substrate: 500 g dw per replicate
- No. of organisms per container (treatment): 10
- No. of replicates per concentration: 8
- No. of replicates per solvent control: 8
- No. of replicates per deionised water control: 4

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition:
5% Sphagnum peat
20% Kaolin-Clay (kaolinite content > 30%)
~74% Quartz sand (fine sand content with particles between 50 and 200 microns higher than 50%)
0.3-1% Calcium carbonate (CaCO3)
- Pretreatment of soil: No
- Stability and homogeneity of test material in the medium: Mixed homogeneously

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark
- Light intensity: Constant intensity at substrate surface: 400 – 800 lx

VEHICLE CONTROL PERFORMED: Yes (acetone in the solvent control)

TEST CONCENTRATIONS
- Range finding study: Not covered by this study plan (previous study)
- Test concentrations: 1000 mg test item/kg soil (dw)
- Results used to determine the conditions for the definitive study: 1000 mg test item/kg soil (dw)

Nominal and measured concentrations:

1000 mg test item/kg soil (dw)

Reference substance (positive control):

yes

Remarks:

Boric acid (separate GLP study; ECT Study No.: IRR1504)

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Biomass

Key result

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Biomass

Duration:

56 d

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality adults at end of exposure period: after 28 days: 5% mortality in control, 7.5% in solvent control and 1.25% in the only concentration of the test item tested.
- Total mass of adults at beginning of test:
Control: 443.6 ± 24.3 mg fresh weight
Solvent control: 444.2 ± 32.6 mg fresh weight
At test concentration: 446.6 ± 29.1 mg fresh weight
- Mean changes in body weigth of live adults (% of initial weight) at end of exposure period:
Control: 54.4
Solvent Control: 52.0
At test concentration: 50.1
- No. of offspring produced (mean ± sd):
Control: 394.3 ± 68.6
Solvent control: 356.4 ± 44.9
At test concentration: 373.0 ± 46.7
- Morphological abnormalities and behavioural abnormalities: No other effects on behavior or morphology of the adult earthworms were observed.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: EC50: 259.1 mg Boric Acid/kg artificial soil (dw)
- Other: 2.5% mortality observed in control, no mortality at the concentrations of 150, 300 and 600 mg Boric acid/kg soil (dw)

Reported statistics and error estimates:

See attachment.

Validity criteria fulfilled:

yes

Remarks:

Mortality of the adults in the solvent control = 7.5% (required: ≤ 10%); Mean number of juveniles in the solvent control = 356.4 (required: ≥ 30); CV for the number of juveniles in the solvent control = 12.6 (required: ≤ 30%).

Conclusions:

Linseed Oil, oxidized did not show any effect on the repoduction and biomass development of earthworm Eisenia fetida. The NOEC mortality, NOECbiomass and the NOECreproduction were at or above 1000 mg test item/kg soil (dw) and accordingly, the LOECmortality, LOECbiomass and the LOECreproduction were above 1000 mg test item/kg soil (dw).

Executive summary:

A 56-day reproduction toxicity test with the earthworm Eisenia fetida in artificial soil was conducted on the substance Linseed Oil, oxidized. The respective guidelines used were the OECD Guideline No. 222 (for the Testing of Chemicals "Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei)") and the ISO Guideline 11268-2 Part 2 (2012) ("Soil Quality – Effects of Pollutants on Earthworms– Part 2: Determination of Effects on Reproduction to Eisenia fetida/Eisenia andrei".)

Except for a short term deviation in form of a temperature drop of 0.3 °C (temperature dropped below 18 °C) no deviations from the test guidelines were recorded during the test. The deviation was considered to have had no impact on the outcome and validity of this study. During the test no abnormal growth or behavioural deviations were recorded. The concentration tested was 1000 mg test item/kg soil (dw). This test concentration did not show any effect on the reproduction and biomass development of the earthworm. Thus, the NOEC for both biomass and reproduction is at or above 1000 mg test item/kg soil (dw).

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Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

a short-term study does not need to be conducted because an appropriate long-term toxicity study on terrestrial organisms is available

Description of key information

No effects were observed in a 56d-study on earthworms (reproduction and biomass) at a concentration of 1000 mg/kg dw. The NOEC is at or above 1000 mg/kg dw

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

Linseed oil, oxidized has a high potential to adsorb to the soil (Log Koc >4.96). In line with Column 2 of Annex IX, a long-term toxicity test was conducted. Therefore, the short term toxicity study with invertebrates is waived.

A 56-day reproduction toxicity test is performed with the earthworm Eisenia fetida in artificial soil according to OECD TG 222 and under GLP. The test is performed as a limit test at 1000 mg test item/kg soil (dw). No abnormal growth or behavioural deviations were recorded nor any effect on the reproduction and biomass development of the earthworm. Thus, the NOEC for both biomass and reproduction is at or above 1000 mg test item/kg soil (dw).