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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Administrative data

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 Aug 2012 - 29 Aug 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
yes
Remarks:
At four days the measured soil temperature increased up to 23.1 °C at maximum for limited time intervals. However, test results have not been impacted.
Qualifier:
according to guideline
Guideline:
EU Method C.8 (Toxicity for Earthworms: Artificial Soil Test)
GLP compliance:
yes (incl. QA statement)
Remarks:
Hessisches Ministerium für Umwelt, Energie, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany

Test material

Constituent 1
Reference substance name:
Glycerides, C14-18 and C16-18-unsatd. mono-, di- and tri-
EC Number:
293-187-5
EC Name:
Glycerides, C14-18 and C16-18-unsatd. mono-, di- and tri-
Cas Number:
91052-28-7
IUPAC Name:
91052-28-7
Details on test material:
- Name of test material (as cited in study report): Glycerides, C14 - C18 and C16 - C18 unsat.
- Substance type: liquid viscous/ yellow
- Physical state: liquid
- Analytical purity: 100 % UVCB
- Lot/batch No.: 0007891181
- Expiration date of the lot/batch: 04 Sep 2013
- Storage condition of test material: stored at ambient temperature
- Water solubility: poorly soluble
- Acetone solubility: soluble (≥ 40 g/L)

Sampling and analysis

Analytical monitoring:
no

Test substrate

Vehicle:
yes
Details on preparation and application of test substrate:
- Method of mixing into soil: The test substance was poorly soluble in water, therefore a volatile organic solvent was used. The stock solution was prepared in acetone. The test substance was dissolved in an amount of acetone sufficient to prepare a stock solution. An appropriate amount of the stock solution was used to soak a portion of quartz sand of the artificial soil. After evaporation of the solvent by placing under a fume hood, the quartz sand was coated with the test substance. The test substance-coated fraction of the quartz sand was then incorporated into the remainder of the artificial soil by thoroughly mixing for approximately 5 min. The respective batches of treated test substrate were mixed by a portable drilling machine equipped with a stirring staff thoroughly to obtain homogeneous distribution of the test substance.
- Controls: blank and solvent control
- Evaporation of vehicle before use: yes, under fume hood

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Earthworm
- Source: The earthworms used in this test were kept at ECT Oekotoxicologie GmbH since February 1994. They were originally delivered by Co. Landenberger (D-72355, Schoemberg)
- Age at test initiation and weight at test initiation: Only adult earthworms (with clitellum) with a fresh weight between 300 and 600 mg were used.

ACCLIMATION
- Acclimation period: Adult earthworms were selected out of the culture and acclimatised for at least 24 h in non-treated artificial soil

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
14 d

Test conditions

Test temperature:
20.0 - 23.1°C
pH:
5.8-6.0
Moisture:
49.5-51.3% of Water Holding Capacity (WHC)
41.1-42.6% of dry mass
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Test vessels were 1.7 L preservation jars, covered by perforated twist-off tops
- Amount of soil: 500 g dry weight per test vessel
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (artificial substrate): The composition of the artificial soil was based on OECD guideline 207 (for percentages refer to dry weight):
10% Sphagnum peat (no visible plant remains; finely ground and air-dried)
20% Kaolin-clay (kaolinite content > 30%)
69-70% Quartz sand (fine sand content with particles between 50 and 200 microns higher than 50%)
0.3-1% Calcium carbonate (CaCO3)
- Maximum water holding capacity (in % dry weigth): 40-60%
- Pretreatment of soil: The air-dried peat was shredded in a chaff-cutter. Afterwards, all parts of the artificial soil were mixed homogeneously. The pH value was adjusted to 6 ± 0.5 using calcium carbonate. The moisture content was adjusted to 40-60% of max WHC using deionised water. The water content of the artificial soil was determined by drying a small sample at 105°C.

OTHER TEST CONDITIONS
- Photoperiod: Permanent
- Light intensity: 523-604 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Number of dead adult earthworms after 7 and 14 d, body weight at the start of the test and after 14 d, morphology and behaviour after 7 and 14 d

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Limit test
Nominal and measured concentrations:
1000 mg/kg artificial soil dry weight (nominal, limit test)
Reference substance (positive control):
yes
Remarks:
Chloroacetamide

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
14 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:
14 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil ww
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: growth, morphology and behaviour
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: growth, morphology and behaviour
Details on results:
- No mortality was observed at the solvent control, blank control and the tested limit concentration of 1000 mg/kg dw
- The biomass of individual adults after 14 d was 90.4% of the initial weight in the solvent control, 91.7% in the control and 96.4% at 1000 mg test substance/kg artificial soil dry weight
- No other symptoms (morphological, behavioural) were observed
Results with reference substance (positive control):
LC50 (14 d) = 42.7 mg/kg (within the validity range given in the ISO Guideline 11268-1 (1997))
Reported statistics and error estimates:
Mortality was not evaluated statistically since no mortality was observed at the limit concentration tested. Biomass was not evaluated statistically since difference in biomass development between control and the limit concentration was 0.9% only.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Under the test conditions, the nominal 14 d NOEC of the test substance to earthworms was >1000 mg/kg dw.
Executive summary:

A study was conducted to determine the short term toxicity of the test substance to earthworms according to OECD guideline 207 and EU method C.8. The test organisms were exposed to a limit concentration of 1000 mg/kg dw of the test substance admixed in soil and the mortality, growth, morphological changes and behaviour of the treated organisms were observed for 14 days post exposure. A solvent control, negative control and a positive control (chloroacetamide) were also kept simultaneously. The LC50 of the positive control was determined to be in the guideline-recommended range. Under the test conditions, the nominal 14 d NOEC of the test substance to earthworms was >1000 mg/kg dw (Moser, 2013).