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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 Aug - 28 Sep 2014
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
according to guideline
other: SEPA, the Guidelines for the Testing of Chemicals, 2004, 207 Earthworm, Acute Toxicity Test.
GLP compliance:
Analytical monitoring:
Details on preparation and application of test substrate:
- Method of mixing into soil: 3000.8 mg of test substances was dissolved in 30mL dichloromethane to give 100 mg/mL solvent stock solution. An aliquot solvent stock solution was added to similar amount of silica sand. The solvent was allowed to evaporate before the incorporation into formulated artificial soil with deionised reverse osmosis water. These were mixed by mixer to give control, solvent control, 1000 mg/kg respectively with a nominal moisture content of 40-60% by mass of the total water-holding capacity with the deionized water.
- Controls: blank control vehicle control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dichloromethane, organic solvent
- Concentration of vehicle in test medium (stock solution and final test solution): 10 mg/kg soil dry weight
- Evaporation of vehicle before use: the solvent was allowed to evaporate before use in artificial soil.
Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
- Common name: earthworms
- Source: Shanghai Yinquan Agriculture Development Co., Ltd., China
- Age at test initiation (mean and range, SD): not stated
- Weight at test initiation (mean and range, SD): 325 - 344 mg

- Acclimation period: 14 days
- Acclimation conditions: breeding on a substrate consisting of 50% peat and 50% cow manure (v/v) and kept under 20 ± 2 °C for 7 days prior to testing.
- Health during acclimation (any mortality observed): not stated
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
Total exposure duration:
14 d
Post exposure observation period:
Test temperature:
20.1 - 21.0 °C
5.81 - 6.01 test initiation
5.92 - 6.05 test end
28.90 - 30.21 test initiation
27.56 - 28.84 test end
Details on test conditions:
- Test container (material, size): 1 L closed glass vessel
- Amount of soil or substrate: 750 g wet weight
- No. of organisms per container (treatment): 40
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4

- Composition of artificial substrate:
Sphagnum peat: 10% (w/w)
Kaolinite clay (Sigma-Aldrich): 20% (w/w)
Silica sand (screened with 0.2 mm sieve): 70% (w/w)
- Organic carbon (%): not stated
- Maximum water holding capacity (in % dry weigth): not stated
- CEC: not stated
- Pretreatment of soil: none
- Storage (condition, duration): not stated
- Stability and homogeneity of test material in the medium: not stated

- Photoperiod: continuous
- Light intensity: 507-611 Lux

mortality after 7 and 14 days


- Range finding study
- Test concentrations: 0, 10, 100, 1000 mg/kg soil dry weight, solvent control (dichloromethane 10 mg/kg dry soil)
- Results used to determine the conditions for the definitive study: LC50 > 1000 mg/kg soil dry weight
Nominal and measured concentrations:
Range finding test: 0, 10, 100, 1000 mg/kg dry soil (nominal)
Limit test: 0 and 1000 mg/kg dry soil (nominal)
Reference substance (positive control):
chloroacetamide, CAS 79-07-2
14 d
Dose descriptor:
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Mortality at end of exposure period in treatment group: 2.5%
- Mortality at end of exposure period in the control group: 2.5%
- Mortality at end of exposure period in the solvent control group: 0%
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: see table 2
- Morphological abnormalities: none
- Behavioural abnormalities: none
Results with reference substance (positive control):
14d LC50 = 11 mg/kg soil dry weight
Reported statistics and error estimates:
Homoscedastic t-Test: differences of the average body weight of the surviving earthworms between the control group and the treatment groups at the end of the study.
Toxcalc software v5.0.26: data analyses

Table 1. Mortality of Eisenia fetida after 7 and 14 days in the limit test:

  Control Solvent control Test substance
1000 mg/kg dry soil
Duration [d] 7 14 7 14 7 14
No. earthworms 40 40 40 40 40 40
No. dead earthworms 1 1 0 0 1 1
Mortality [%] 2.5 2.5 0 0 2.5 2.5

Table 2. Average body weight of the surviving earthworms in the limit test:

  Replicate No. earthworms
Day 0
Mean weight
Day 0
No. earthworms
Day 14
Mean weight
Day 14
Weight reduction after 14 days [% ± SD]
Control A 10 329 9 316 5.56%± 1.72
B 10 344 10 322
C 10 344 10 318
D 10 325 10 311



A 10 341 10 324 12.45% ± 6.61
B 10 368 10 295
C 10 351 10 296
D 10 326 10 296
Test substance
1000 mg/kg dry soil
A 10 344 9 316 18.87% ± 11.66
B 10 391 10 254
C 10 346 10 300
D 10 400 10 324

Description of key information

LC50 (14 d) > 1000 mg/kg dry soil (OECD 207)

Key value for chemical safety assessment

Additional information

The short term effects of Reaction mass of 1,1'-(isopropylidene)bis[3,5-dibromo-4-(2,3-dibromo-2-methylpropoxy)benzene] and 1,3-dibromo-2-(2,3-dibromo-2-methylpropoxy)-5-{2-[3,5-dibromo-4-(2,3,3-tribromo-2-methylpropoxy)phenyl]propan-2-yl}benzene to soil macroorganisms were investigated in a study by Jing (2014) according to OECD guideline 207 and GLP. The test was performed with the annelid Eisenia fetida as the test organism and artificial soil. Dichloromethane was used as a vehicle. The test organism was exposed to a single nominal concentration of 1000 mg/kg dry soil for 14 d. At test termination after 14 d 2.5% mortality was observed in the control and in the test substance treatment but not in the solvent control. The resulted LC50 (14 d) was > 1000 mg/kg dry soil.