1-Ethoxy-2,3-difluor-4-[4-(trans-4-propylcyclohexyl)-1-cyclohexen-1-yl]-benzene

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

Study period:

Aug 10, 2010 - Dec 17, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

other: Guidelines for the testing of chemicals (HJ/T153-2004) and Guidelines for the testing of chemicals No. 207, China

Deviations:

not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

not specified

Principles of method if other than guideline:

Due to the limited water solubility (below 100 mg/L) the test was conducted under Chinese regulation at 100 mg/kg soil.

GLP compliance:

no

Remarks:

no OECD GLP

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

A defined artificial soil of the following composition was used for this assay.

Sphagnum Peat 10 % (pH 5.5-6.0, no visible plant remains)
Kaolin Clay 20 % (kaolinite content preferably above 30 %)
Industrial Sand 68 % (fine sand should be dominant with more than 50 % of the particles between 50-200 μm)
CaC03 2 % (AR)

The moisture content of all constituents above was determined by drying a small sample at 105 °C and re-weighing and then blended in the correct proportions and mixed thoroughly. Deionised water was added, and the medium was thoroughly mixed. The test substance was mixed directly with artificial soil by motor stirring machine. A high concentration mixture of test substance and dry artificial soil was used as the test stock soil.
Test substance stock soil with various volumes was mixed thoroughly with artificial soil to prepare a set of test medium.
750 g (wet weight) of test medium placed into each container, with ten earthworms each. Those earthworms, that were washed with distilled water quickly before use, are placed on the test medium surface. The containers are covered with perforated plastic film to prevent the test medium from drying.

Test organisms (species):

Eisenia fetida

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida
- Source: Jialiming Earthworm Breeding Farm in Tianjin, China
- Age at test initiation: 3 months
- Weight at test initiation: 0.29 +/- 0.04 g

ACCLIMATION
- Acclimation period: 7 days (14 days without medication)
- Acclimation conditions: same as test
- Health during acclimation (any mortality observed): no mortality, no other findings

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

20 +/- 22 °C

pH:

7.52 - 7.84

Moisture:

34.9 %

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 1 L glass vessel, closed
- Amount of soil or substrate: 750 g
- 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

- Composition of soil: Please refer to "Details on preparation and application of test substrate".

OTHER TEST CONDITIONS
- Photoperiod: 24 h/day
- Light intensity: 573 - 581 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
day 7 and 14

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
A range finding study using test concentrations 0, 0.1, 1.0, 10 and 100 mg/kg dry weight of artificial soil using 2 replicates per concentration was performed. The duration was 7 days, temperature 20 +/- 2 °C and humidity 80 % with continuous light at 4000-6000 Lux.
- Results used to determine the conditions for the definitive study: yes

Nominal and measured concentrations:

100 mg/kg soil

Reference substance (positive control):

yes

Remarks:

Chloroacetamide

Duration:

7 d

Dose descriptor:

LC50

Effect conc.:

> 100 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

14 d

Dose descriptor:

LD50

Effect conc.:

> 100 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

The mortality of the blank control group on day 7 was 0 %. The mortality of the treatment group on day 7 was 2.5 %. The mortality of control group on day 14 was 2.5 % and 10 % for the treatment group.

Results with reference substance (positive control):

The 24 h LC50 of chloroacetamide was calculated as 3.3 * 10^-3 mg/cm^2 (validity crititeria: 1.5 to 4.5 * 10^-3 mg/cm^2).

Reported statistics and error estimates:

Standard statistical methods have been applied for data processing.

Validity criteria fulfilled:

yes

Conclusions:

The 7-day LD50 and the 14-day LD50 for the test substance to earthworms (Eisenia fetida) based on nominal test concentrations were higher than 100 mg/kg dry soil weight.

Executive summary:

In this study, the acute toxicity of the test substance to Eisenia fetida was determined in a 14 days artificial soil test. The study design was similar to OECD 207 and according to guidelines for the testing of chemicals (HJ/T153-2004) and guidelines for the testing of chemicals No. 207, China. Based on the results of range-finding test, one treatment group and one blank control group, which had four replicates of 10 worms per concentration each were included in a limit test. The nominal concentration of the treatment group was 100 mg/kg dry artificial soil weight. The number of mortalities were determined after 7 and 14 days of exposure. Mortality of the earthworms in blank control group was 2.5 % after 14 days, which was not more than 10 %, so the test was valid. The mortality of the treatment group on day 7 was 2.5 % and after 14 days 10 %. The 24 h LC50 of the positive control substance chloroacetamide was calculated as 3.3 * 10^-3 mg/cm^2, thus meeting the validity crititeria of 1.5 to 4.5 * 10^-3 mg/cm^2. Under conditions of the test, the test substance had no acute toxicity effects on the earthworm. The 7-day LD50 and the 14-day LD50 for the test substance to earthworms (Eisenia fetida) based on nominal test concentrations were higher than 100 mg/kg dry soil weight.

Description of key information

The 7-day LD50 and the 14-day LD50 for the test substance to earthworms (Eisenia fetida) based on nominal test concentrations were higher than 100 mg/kg dry soil weight.

Additional information

In this study, the acute toxicity of the test substance to Eisenia fetida was determined in a 14 days artificial soil test. The study design was similar to OECD 207 and according to guidelines for the testing of chemicals (HJ/T153-2004) and guidelines for the testing of chemicals No. 207, China. Based on the results of range-finding test, one treatment group and one blank control group, which had four replicates of 10 worms per concentration each were included in a limit test. The nominal concentration of the treatment group was 100 mg/kg dry artificial soil weight. The number of mortalities were determined after 7 and 14 days of exposure. Mortality of the earthworms in blank control group was 2.5 % after 14 days, which was not more than 10 %, so the test was valid. The mortality of the treatment group on day 7 was 2.5 % and after 14 days 10 %. The 24 h LC50 of the positive control substance chloroacetamide was calculated as 3.3 * 10^-3 mg/cm^2, thus meeting the validity crititeria of 1.5 to 4.5 * 10^-3 mg/cm^2. Under conditions of the test, the test substance had no acute toxicity effects on the earthworm. The 7-day LD50 and the 14-day LD50 for the test substance to earthworms (Eisenia fetida) based on nominal test concentrations were higher than 100 mg/kg dry soil weight.