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

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:

14 Aug - 28 Sep 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SEPA, the Guidelines for the Testing of Chemicals, 2004, 207 Earthworm, Acute Toxicity Test.

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

yes

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:

annelids

Details on test organisms:

TEST ORGANISM
- 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
- 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:

yes

Total exposure duration:

14 d

Post exposure observation period:

none

Test temperature:

20.1 - 21.0 °C

pH:

5.81 - 6.01 test initiation
5.92 - 6.05 test end

Moisture:

28.90 - 30.21 test initiation
27.56 - 28.84 test end

Details on test conditions:

TEST SYSTEM
- 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

SOURCE AND PROPERTIES OF SUBSTRATE
- 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

OTHER TEST CONDITIONS
- Photoperiod: continuous
- Light intensity: 507-611 Lux

EFFECT PARAMETERS MEASURED:
mortality after 7 and 14 days

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- 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):

yes

Remarks:

chloroacetamide, CAS 79-07-2

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

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 weight
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
Solvent control	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 weight	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)

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.