(E)-anethole

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Version / remarks:

1984

GLP compliance:

yes (incl. QA statement)

Vehicle:

yes

Remarks:

Acetone

Details on preparation and application of test substrate:

The respective test item amount was weighed out for each test item concentration and dissolved in acetone (2.5 mL per replicate). The appropriate amount of spiking solution was mixed thoroughly with quartz sand (10 g per replicate). After complete evaporation of the solvent, the spiked quartz sand was added to the artificial soil.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: Breeding stock culture maintained at the test facility
- Age at test initiation (mean and range, SD): aged 2- 12 month, with clitellum
- Weight at test initiation (mean and range, SD): 0.30 - 0.64 g/worm

ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions: same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

2 wk

Test temperature:

18-22 °C

pH:

Day 0: 5.76 - 5.84
Day 14: 5.74 - 5.84

Moisture:

Day 0: 27.7 - 28.3 % of DW
Day 14: 26.2 - 27.3 % of DW

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Glass vessels, volume 1.5 L
- Amount of soil or substrate: 555 g soil dw
- 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

OTHER TEST CONDITIONS
- Photoperiod: 24 h
- Light intensity: 595 lx +- 108 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality, behaviour and pathological symtoms after 7 and 14 days

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline:
- Range finding study: yes
- Test concentrations: 1, 10, 100, 1000 mg/kg soil dw
- Results used to determine the conditions for the definitive study: 100% mortality in the 1000 mg/kg soil dw conc. and 0% Mortalöity in the 100 mg/kg soil dw conc.

Nominal and measured concentrations:

control, solvent control, 50, 100, 200, 400, 800 mg/kg soil dw

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

Key result

Duration:

2 wk

Dose descriptor:

LC50

Effect conc.:

383 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

2 wk

Dose descriptor:

NOEC

Effect conc.:

200 mg/kg soil dw

95% CI:

383 - 432

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Motality, behaviour, pathological symtoms, biomass

Details on results:

- Mortality at end of exposure period: No mortality was observed in the controls and the test item concentrations 50 to 200 mg/kg SDW after 14 days of exposure. However, at the test item concentrations 400 and 800 mg/kg SDW a
statistically significant mortality of 57.5 % and 100 % occurred, respectively.
- Changes in body weight of live adults (% of initial weight) at end of exposure period: s. table 2
- Behavioural abnormalities and pahological symtoms: No pathological symptoms or changes in earthworm behaviour was observed in the controls and the test item concentrations 50 to 200 mg/kg SDW after 14 days of exposure. At the test item concentration 400 mg/kg SDW some earthworms showed spontaneous segmentation and
separation after 7 and 14 days of exposure

Results with reference substance (positive control):

LC50 (2 wk) = 44.5 mg/kg soil dw

Reported statistics and error estimates:

- Mortality / Biomass: The arithmetic mean and NOEC were determined.- Dunnett’s Multiple Comparison Test

- LC50-values and confidence ranges for mortality: The LC50-value in terms of mortality was determined by Sigmoidal dose-response (variable slope).

Table 1: Earthworm Mortality in [%] after 7 and 14 Days of Exposure

	Mortality [%]										Statistically
Test item concentration	Day 7					Day 14					significant vs.
[mg/kg SDW]	1	2	3	4	MV	1	2	3	4	MV	Pooled Control*
Control	0	0	0	0	0	0	0	0	0	0	-
Solvent Control	0	0	0	0	0	0	0	0	0	0	-
Pooled Control	-				0	-				0	-
50	0	0	0	0	0	0	0	0	0	0	No
100	0	0	0	0	0	0	0	0	0	0	No
200	0	0	0	0	0	0	0	0	0	0	No
400	40	20	70	60	47.5	40	40	90	60	57.5	Yes
800	100	100	100	100	100	#					Yes

 

Table 2: Mean Live Weight and Mean Loss of Body Weight

	Mean live weight
Test item	Test start	Test end	Inhibition	Statistically
concentration	MV ± SD	MV ± SD	[%]	significant vs.
[mg/kg SDW]	[g/worm]	[g/worm]		Pooled Control
Control	0.48 ± 0.05	0.41 ± 0.04	15	–
Solvent control	0.48 ± 0.04	0.42 ± 0.05	13	-
Pooled Control	0.48 ± 0.04	0.42 ± 0.05	14	-
50	0.48 ± 0.06	0.42 ± 0.05	13	No
100	0.48 ± 0.06	0.40 ± 0.05	17	No
200	0.48 ± 0.05	0.40 ± 0.05	17	No
400	0.48 ± 0.06	0.38 ± 0.08	21	Yes
800	0.48 ± 0.05	#

Validity criteria fulfilled:

yes

Conclusions:

The LC50 (2 wk) of the test item to earthworm (Eisenia fetida) was determined to be = 383 mg/kg soil dry weight.

Executive summary:

The acute effects of the test item on the earthworm Eisenia fetida were determined according to OECD Guideline 207 according to GLP. The study was conducted under static conditions for 14 days with the test item concentrations 50 – 100 – 200 – 400 – 800 mg/kg SDW mixed into artificial soil containing 10 % peat. Artificial soil without test item was used as control and artificial soil without test item but treated with the solvent was used as solvent control. 40 test organisms divided into four replicates, were tested per test item concentration, control and solvent control. No statistically significant differences concerning mortality and biomass after two weeks of exposure in soil were observed between the control and the solvent control.  After 14 days, the EC50 of the test item to earthworm (Eisenia fetida) was determined to be 383 mg/kg soil dry weight.

Description of key information

Based on results of an OECD 207 compliant study with the test item an EC50 of 383 mg/kg soil dw was determined as key value for the risk assessment.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

383 mg/kg soil dw

Additional information

The acute effects of the test item on the earthworm Eisenia fetida were determined according to OECD Guideline 207 according to GLP. The study was conducted under static conditions for 14 days with the test item concentrations 50 – 100 – 200 – 400 – 800 mg/kg SDW mixed into artificial soil containing 10 % peat. Artificial soil without test item was used as control and artificial soil without test item but treated with the solvent was used as solvent control. 40 test organisms divided into four replicates, were tested per test item concentration, control and solvent control. No statistically significant differences concerning mortality and biomass after two weeks of exposure in soil were observed between the control and the solvent control.  After 14 days, the EC50 of the test item to earthworm (Eisenia fetida) was determined to be 383 mg/kg soil dry weight.