Reaction mass of (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)butan-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)butan-2-one

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:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals, Effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test[M] (2013)

Version / remarks:

The test was conducted in accordance with China HJ/T153 - 2004 Chemical Test Guideline stipulations and Chemical Registration Center of MEP. The Guidelines for the Testing of Chemicals, effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test[M]. Second edition. Beijing: China
Environmental Press. 2013: 58-62. The test was also completed under OECD TG 207: Earthworm, Acute Toxicity (1984).

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: China National Technical Committee for Standardization of Dangerous Chemical Management. GB/T 21809-2008, Chemical Earthworm, Acute Toxicity Tests [S]. Beijing: China Standardization Press (2008)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): The density of substance was reported as 0.930 mg/μL. 10g of a mixture of fine ground quartz sand was mixed with 60 μL, 106 μL, 189 μL, 336 μL and 597 μL mg test substance respectively and added to 740 g wet artificial soil to form the test substance concentrations of 100 mg/kg dry soil, 178 mg/kg dry soil, 316mg/kg dry soil, 562 mg/kg dry soil and 100 mg/kg dry weight. Four replicates were prepared for each test group by the method.
- Controls: The blank (negative) control group consisted of a test system without application of the test item.

Test organisms (species):

Eisenia sp.

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Earth worm.
- Source: Zhuhai Hongming Earthworm Breeding Park (China)
- Age at test initiation (mean and range, SD): older than 2 months.
- Weight at test initiation (mean and range, SD): mean weight 0.516 – 0.532 g. Individual weight of 0499 g – 0.545 g; Individual test item concentration groups – mean and individual weights are reported in the full study report.

ACCLIMATION
- Acclimation period: kept in the artificial soil for 24h before start of the test
- Acclimation conditions (same as test or not): Yes.
- Health during acclimation (any mortality observed): None reported; organisms were reported as breeding in the artificial soil in the 24 hours prior to the test in the non-GLP premliminary soil test.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Remarks:

Sorting the Earthworms from the artificial soil and testing their reaction to a mechanical stimulus at the front end at 7d. The death condition were checked and recorded. Any behavioural or pathological symptoms were checked and recorded.

Test temperature:

Preliminary study (non-GLP): 19.2 °C - 22.0 °C
Definitive study: 18.1 °C - 21.8 °C

pH:

pH: pH of the artificial soil was measured at the beginning of the test. Weighed 5 g of air-dried soil sample in 50 mL beaker. 25 mL CO2- free distilled water was added by cylinder. Stirred for 5 min. Standing for 1 h and then measurement of the pH. pH of the artificial soil at the start of the test:
Temperature: 21.8 °C ; First reading pH: 6.46; Second reading pH: 6.46 and Mean pH: 6.46

Moisture:

Moisture of the artificial soil was measured at the beginning and at the end of the test. The average of three weighings was calculated.
Start: Mean moisture range per test group: 24.7% - 25.9%;
Day 14 - end: Mean moisture range per test group: 23.4% - 23.8%

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 1 Litre; glass
- Amount of soil or substrate: 740 g wet artificial soil
- No. of organisms per container (treatment):
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: Not applicable.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Not applicable.
- Sampling depth (cm):
- Composition (if artificial substrate):
- % sand: 70% industrial sand (particle size between 40-100 μm)
- % silt: sphagnum peat 10% (no visible plant remains)
- % clay: 20% kaolin clay ((AR, Weight loss on ignition< 15%)
- Organic carbon (%): not reported
- Maximum water holding capacity (in % dry weight): Deionized water was added to give an overall moisture content of 35% of the dry weight, and the medium was thoroughly mixed
- Pretreatment of soil: The dry constituents were blended in the correct proportions Moisture content was then determined by drying a small sample at 1 05°C ±5 °C and re-weighing. Deionized water was added to give an overall moisture content of 35% of the dry weight, and the medium was thoroughly mixed. The pH was measured as close to pH 5.5 to 6.5 as possible.
- Storage (condition, duration): Not applicable. Artificial soild prepared prior to exposure.
- Stability and homogeneity of test material in the medium: Stable. No instability reported.

OTHER TEST CONDITIONS
- Photoperiod: Continuous light
- Light intensity: The light intensity was 600 lx at the beginning of the test and 610 lx at the end of the test.
- Other: The humidity was kept constant by the humidifier from 67.0% to 100% during the test period.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: No.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Definitive study: 0 (blank control), 100, 178, 316, 562 and 1000 mg/kg dry weight (nominal)
- Justification for using less concentrations than requested by guideline: Not applicable.
- Range finding study
- Test concentrations: Preliminary study (non-GLP): 0 (blank control), 0 (solvent control), 1.0, 10, 100, 1000 mg/kg dry weight (nominal)
- Results used to determine the conditions for the definitive study: Yes.

Nominal and measured concentrations:

Preliminary study (non-GLP): 0 (blank control), 0 (solvent control), 1.0, 10, 100, 1000 mg/kg dry weight (nominal)
Definitive study: 0 (blank control), 100, 178, 316, 562 and 1000 mg/kg dry weight (nominal)

Reference substance (positive control):

yes

Remarks:

chloracetamide

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

152 mg/kg soil ww

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% C.I. = 130 - 174 mg/kg dry soil

Results with reference substance (positive control):

- Results with reference substance valid? Yes.
- Relevant effect levels: 18.3 (C.I. 17.5 - 19.1) mg/kg
- Other: Not applicable.

Reported statistics and error estimates:

Mortality/concentration data are recorded and the median lethal concentration (LC50) and its 95% confidence limits estimated using a Probit Analysis Programme.

Test 1. Mortality during the artificial soil test

Group	Number of earthworms	7d			14d
		Mortalities	Mortality (%)	Modified mortality (%)	Mortalities	Mortality (%)	Modified mortality (%)
Blank control	40	0	0	0	0	0	0
100 mg/kg dry soil	40	7	17.5	17.5	11	27.5	27.5
178 mg/kg dry soil	40	14	35	35	20	50	50
316 mg/kg dry soil	40	37	92.5	92.5	38	95	95
562 mg/kg dry soil	40	40	100	100	40	100	100
1000 mg/kg dry soil	40	40	100	100	40	100	100

Note:

The modified mortality (%) = (Number of mortalities in treatment group - Number of mortalities in control group) / (Number treatment group - Number in control group) x 100%

 

All earthworms in blank control were under the surface of the soil and healthy throughout the exposure period (7d and 14d). No other significant pathological symptoms or abnormal behaviour (other than mortality) were reported in any treatment groups.

Validity criteria fulfilled:

yes

Conclusions:

The test substance 14d LC50 was determined to be 132 (C.I. 130 - 174) mg/kg dry soil based on nominal loading

Executive summary:

The acute toxicity of the test item to earth worm (Eisenia foetida) was determined in a 14d acute toxicity test in artificial soil according to OECD TG 207 and the China CRC MEP guidelines, Effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test (2013) under GLP. Following a non-GLP preliminary test a definitive test of five test groups of 0 (blank control), 100, 178, 316, 562 and 1000 mg/kg dry weight (nominal) were conducted with four replicates per group in artificial sand. The test substance was mixed in a 10 g of a mixture of fine ground quartz sand, the test item at defined dry weight nominal concentration to 740 g wet artificial soil to form the test substance concentrations. The test temperature was maintained at 18.1 - 21.8 °C under continuous light at 600 lux. The test system was validated against a chloracetamide reference item. There was no mortality in the blank control group and no sign of abnormal behaviour. The 14d LC50 was determined to be 132 (C.I. 130 - 174) mg/kg dry soil based on nominal loading. All relevant guideline criteria were met.

Description of key information

LC50 (earthworm) = 152 (C.I. 130 -172) mg/kg dry weight nominal, 14 day, OECD TG 207, 2015

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

152 mg/kg soil dw

Additional information

The acute toxicity of the test item to earth worm (Eisenia foetida) was determined in a 14d acute toxicity test in artificial soil according to OECD TG 207 and the China CRC MEP guidelines, Effects on Biotic Systems, 207 Earthworm, Acute Toxicity Test (2013) under GLP. Following a non-GLP preliminary test a definitive test of five test groups of 0 (blank control), 100, 178, 316, 562 and 1000 mg/kg dry weight (nominal) were conducted with four replicates per group in artificial sand. The test substance was mixed in a 10 g of a mixture of fine ground quartz sand, the test item at defined dry weight nominal concentration to 740 g wet artificial soil to form the test substance concentrations. The test temperature was maintained at 18.1 - 21.8 °C under continuous light at 600 lux. The test system was validated against a chloracetamide reference item. There was no mortality in the blank control group and no sign of abnormal behaviour. The 14d LC50 was determined to be 132 (C.I. 130 - 174) mg/kg dry soil based on nominal loading. All relevant guideline criteria were met.