N,N'-bis{3-[di-(branched and cyclic) alkylamino]alkyl}alkanediamide

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:

Sep 24, 2014 to Jan 28, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

GLP compliance:

yes

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Stability under test conditions: stable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The 5.603g test substances dissolved in 50 mL chloroform, configured for 112 g/L by using the liquid reserves. A certain amount of the stock solution with 10g quartz sand mixing, volatile dry chloroform. A total amount of 10g 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.
- Final preparation of a solid: The medium was thoroughly mixed and deionised water was added finally to give an overall moisture content of about 35 per cent of the dry weight. Moisture content was determined by drying a small sample at 105°C and re-weighing. The complete mixture was moist but not so wet that water appears when the artificial soil is compressed.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1000 mg/kg (dry soil)

Vehicle:

yes

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name:
- Source: The worms were obtained from an · earthworm nursery of Nanjing city in China. The earthworms newly obtained in the lab will be released from quarantine for 14 days and then this batch was turned into pre-culture.
- Age at test initiation (mean and range, SD): Worms for this study were adult (two months old with clitellum)
- Weight at test initiation (mean and range, SD): with an individual weight of 346 mg~ 375 mg and the average weight of 362 mg.

ACCLIMATION
- Acclimation period: 53 days
- Acclimation conditions (same as test or not): same

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

20.8°C to 22.2°C

pH:

6.1 to 6.3

Moisture:

In range of 27% to 35%

Details on test conditions:

TEST SYSTEM
- Test container (material, size):
- Amount of soil or substrate: 560g
- No. of organisms per container (treatment): 10 worms each
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 4

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

VEHICLE CONTROL PERFORMED: yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: NA
- Range finding study: A pre-test was performed with test substance concentrations of 10.0, 100 and 1000 mg/kg artificial soil (dry weight). Simultaneously one blank control and one solvent control were stablished under the same conditions.
- Results used to determine the conditions for the definitive study: Based on the results of the range-finding test, a Limit Test was undertaken with test substance concentration of 1000 mg/kg (dry weight of soil), a blank control and solvent control.

Nominal and measured concentrations:

1000 mg/kg (dry weight of soil)

Reference substance (positive control):

yes

Key result

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

Key result

Duration:

14 d

Dose descriptor:

LC0

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: no effect
- Changes in body weight of live adults (% of initial weight) at end of exposure period: After the test, weight reductions of the blank control, solvent control and test group were 52 mg - 56 mg, 52 mg - 55 mg and 52 mg - 53 mg, respectively.
- Behavioural abnormalities: At the beginning of the administration, test worms soon disappeared into the test soil medium. During the test, all earthworms remained in the soil.

Reported statistics and error estimates:

The LC50 and 95% confidence limits can be used to calculate by Trimmed Speannan-Karber Method (Version 1.5, USEPA).
The body weight test data was analyzed by analysis of variance (ANOVA, Dunnett's tests, SPSS 17.0)

Validity criteria fulfilled:

yes

Conclusions:

The 14 d LC50 of the test substance in the artificial soil was determined to be greater than 1000 mg/kg dry weight soil.

Executive summary:

The test was conducted to determine the acute tox1c1ty of test substances to earthworm according to "The guidelines for the testing of chemicals" (HJ/T 153-2004) and "The Guidelines for the Testing of Chemicals-Effects on Biotic Systems" (the second edition. Beijing: China Environmental Science Press, 2013); while with reference to Procedure 207 of the 'Guidelines for Testing of Chemicals' of the OECD: "Earthworm, Acute Toxicity Tests" (1984) etc.

During this test, the artificial soils was used with the moisture content of the test medium controlled in range of 27% to 35% with pH from 6.1 to 6.3 The test conditions were maintained as follows: temperature of 20.8 °C to 22.2 °C, humidity of 80% to 85% and continuous light with intensity of 400 lux to 800 lux.

Based on the result of range-finding test and the Limit Test, the 14 d LC50 of the test substance in the artificial soil was determined to be greater than 1000 mg/kg dry weight soil. The highest concentration causing no mortality (14 d-LC0) was 1000 mg/kg dry weight soil.

Description of key information

The test was conducted to determine the acute toxicity of test substances to earthworm according to "The guidelines for the testing of chemicals" (HJ/T 153-2004) and "The Guidelines for the Testing of Chemicals-Effects on Biotic Systems" (the second edition. Beijing: China Environmental Science Press, 2013); while with reference to Procedure 207 of the 'Guidelines for Testing of Chemicals' of the OECD: "Earthworm, Acute Toxicity Tests" (1984) etc.

During this test, the artificial soils was used with the moisture content of the test medium controlled in range of 27% to 35% with pH from 6.1 to 6.3 The test conditions were maintained as follows: temperature of 20.8 °C to 22.2 °C, humidity of 80% to 85% and continuous light with intensity of 400 lux to 800 lux.

Based on the result of range-finding test and the Limit Test, the 14 d LC50 of the test substance in the artificial soil was determined to be greater than 1000 mg/kg dry weight soil. The highest concentration causing no mortality (14 d-LC0) was 1000 mg/kg dry weight soil.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Long-term EC10, LC10 or NOEC for soil macroorganisms:

1 000 mg/kg soil dw