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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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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:
Mar 3, 2015 to Jul 17, 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Version / remarks:
1984
Qualifier:
according to guideline
Guideline:
other: HJ/T 153-2004. The guidelines for the testing of chemicals [S]. Beijing: State Environmental Protection Administration, 2004
Qualifier:
according to guideline
Guideline:
other: CRC-MEP. The Guidelines for the Testing of Chemicals-Effects on Biotic Systems. [M]. The second edition. Beijing: China Environment Press. 2013: 58-62
Qualifier:
according to guideline
Guideline:
other: GBff 21809-2008, Chemicals-Test method of earthworm acute toxicity test [S]. Beijing: Standardization Administration of China, 2008
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method C.8 (Toxicity for Earthworms: Artificial Soil Test)
Version / remarks:
2008
GLP compliance:
yes
Remarks:
According to the guidelines of chemical testing good laboratory practices in China
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch No. 6Fl 1027000
- Expiration date of the lot/batch: 2 years
- Purity: 96%
Analytical monitoring:
no
Vehicle:
yes
Remarks:
Acetone (in the definitive test)
Details on preparation and application of test substrate:
- Method of mixing into soil:
FOR THE RANGE FINDING TEST: the test substance was directly added into the soil. A total amount of 10 g mixture of the test substance and quartz sand was prepared firstly and then mixed with 550 g dry weight of soil for each test container. The medium was thoroughly mixed and deionised water was added finally to give an overall moisture content of about 35% dw.
FOR THE DEFINITIVE TEST: A stock solution was prepared be dissolving 5.2088 g test substance into 50.0 mL acetone. Test substrates at different concentrations were prepared by adding the stock solution to 10 g quartz sand and mixed. After evaporating the acetone at the room temperature, 10 g quartz sand then mixed with 550 g dry weight of soil for each test container. The medium was thoroughly mixed and deionised water was added finally to give an overall moisture content of about 35% dw.
- Evaporation of vehicle before use: For the definitive test, the acetone was evaporated at room temperature
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: An earthworm nursery of Nanjing city in China (Batch No.: E20150327E).
- Age at test initiation (mean and range, SD): Not specified
- Weight at test initiation (mean and range, SD): Range of average weight was 332-421 mg

ACCLIMATION
- Acclimation period: before the test, worms were acclimated in test soil for 17 days. Worms were then pre-cultured in test soils for another 24hrs. Worms were washed quickly with distilled water and surplus water absorbed on filter paper before use
- Acclimation conditions (same as test or not): Not reported (Noting that worms were acclimated in test soils for 24hrs)
- Health during acclimation (any mortality observed): Not specified
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
14 d
Remarks:
14 days duration for range finding test and 14 days duration for definitive test
Post exposure observation period:
Mortality assessed on 7d and 14 d; number and type of behavioural and other pathological abnormalities (e.g. lethargic or rigid worms) were determined during each mortality assessment at 7d and 14 d.
Test temperature:
20.5°C to 21.8°C
pH:
6.10 - 6.39
Moisture:
27% to 35%
Details on test conditions:
TEST SYSTEM
- Amount of soil or substrate: dry weight: 560 g each
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: For the definitive test, 4 replications for each concentration level
- No. of replicates per control: 4
- No. of replicates per solvent control: 4

SOURCE AND PROPERTIES OF SUBSTRATE
- Artificial soil:
10% sphagnum peat
20% kaolin clay
70% quartz sand

OTHER TEST CONDITIONS
- Photoperiod: continuous light
- Light intensity: 400 lux to 800 lux

EFFECT PARAMETERS MEASURED: mortality and behavioral effects; observations taken at 7 and 14 days

VEHICLE CONTROL PERFORMED: Yes (solvent control)

TEST CONCENTRATIONS
- Definitive test concentrations: 100, 150, 225, 338, 506 mg/kg artificial soil (dry weight of soil)
- Spacing factor for test concentrations: 1.5

- Range finding study
- Test concentrations: 100, 500 and 1000 mg/kg artificial soil (dry weight)
- Results used to determine the conditions for the definitive study: Yes
Nominal and measured concentrations:
100, 150, 225, 338, 506 mg/kg artificial soil (dry weight of soil)
Reference substance (positive control):
yes
Remarks:
2-Chloracetamide, CAS: 79-07-2; purity: 2: 98.5%; Lot number: 20130905; Sinopharm Chemical Reagent Co. Ltd.
Key result
Duration:
7 d
Dose descriptor:
LC50
Effect conc.:
184 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence 174-194 mg/kg dw soil
Duration:
7 d
Dose descriptor:
NOEC
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:
LC50
Effect conc.:
150 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% confidence 141 to 160 mg/kg dw soil
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Results with reference substance (positive control):
The 7-days LC50 and 14-days LC50 for 2-chloracetamide were 29.3 mg/kg and 21.2 mg/kg, respectively.

The 7-day and 14-day LC50 values for 2-chloracetamide are determined 3 times a year to assure that the laboratory test conditions are adequate and had not changed significantly.
Reported statistics and error estimates:
The LC50 and 95% confidence intervals were used to calculate by Trimmed Spearman-Karber Method (Version 1.5, USEPA).

Summary of validation criteria:

(1)         Control group

A blank control group, comprising the same number of test worms as each treatment, was also treated with the same methods while without using the test substance.

(2)         Reference control

The 7-day and 14-day LC50 values of a reference substance (2-chloracetamide) were determined 3 times a year to assure that the laboratory test conditions were adequate and had not changed significantly. The 7-days LC50 and 14-days LC50 for reference substance were 29.3 mg/kg and 21.2 mg/kg, respectively.

(3)         Validity of test result

Mortality in the untreated control group at the end of the study must not exceed 10%. Otherwise the test should be considered invalid.

Validity criteria fulfilled:
yes
Conclusions:
The test was conducted to determine the acute toxicity of the test item to earthworm. Based on the result of range-finding test and definitive test, the 7-d and 14-d LC50 of the test item in artificial soil were determined to be 184 mg/kg dw soil and 150 mg/kg dw soil, respectively. The highest concentration causing no mortality 7 day and 14 day LC0 were 100 mg/kg dry weight soil.
Executive summary:

The acute toxicity of 1,3,4-triisopropylbenzen was tested under static conditions in a 14-day toxicity study on the earthworm (Eisenia fetida) in a study similar to OECD TG 207. Based on nominal concentrations, the 7-day LC50 was determined to be 184 mg/L with a NOEC of 100 mg/L based on mortality. The 14-day nominal LC50 was determined to be 150 mg/L with a NOEC of 100 mg/L based on mortality.

Description of key information

The acute toxicity of 1,3,4-triisopropylbenzen was tested under static conditions in a 14-day toxicity study on the earthworm (Eisenia fetida) in a study similar to OECD TG 207. Based on nominal concentrations, the 7-day LC50 was determined to be 184 mg/L with a NOEC of 100 mg/L based on mortality. The 14-day nominal LC50 was determined to be 150 mg/L with a NOEC of 100 mg/L based on mortality.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:
150 mg/kg soil dw

Additional information