Reaction products of N,N'-ethane-1,2-diylbis(1,3-propanediamine), cyclohexane, peroxidized 4-butylamino-2,2,6,6-tetramethylpiperidine and 2,4,6-trichloro-1,3,5-triazine

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:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

Based on the specified water solubility the application of the test substance via an aqueous stock solution is not possible. The water solubility is not enough to prepare a sufficient stock solution with deionized water. Application of the test substance: ground in a mortar and distributed in quartz sand and added to the test substrate afterwards. The required amounts of test substance were weighed into a beaker and mixed with 25 g quartz sand.

Preparing the control assays:
For preparation of the test assays of the control, the amount of about 2535 g dry test was added to a bowl. The untreated quartz sand aliquot was added after and thoroughly mixed with the test substrate. Than the required volume of 715 mL deionized water was added to the test substrate, respectively and mixed well again. Four portions of 750 g of wet substrate were distributed to preserving jars.

Preparing the test substance assays:
For preparation of the test assays with test substance amounts of about 2535 g dry test substrate was added to a box. The quartz sand aliquots treated with the required test substance amounts were added and thoroughly mixed with the test substrate. Than the required volumes of 715 mL deionized water was added to the test substrate test substance mixtures and mixed well again. Four portions of 750 g of the treated wet test substrate for each test concentration were distributed to preserving jars.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Species: Earthworm Eisenia Fetida
Origin: ECT, Ecotoxicology GmbH, Böttgerstr. 2-14, D-65349 Flörsheim, Germany
Feeding of the earthworms: The worms were feed with oat flakes at least every second week.
Age of the animals at the start of exposure: approx. 6 months (guaranteed by the supplier)
Test animals: Animals with a weight of 325 mg – 421 mg (calculated values from the mean values of replicates) and a clear visible clitellum were used for the test
Conditions of storage at the test facility: The worms were stored in the transport box until adaptation at room temperature in the dark.
Adaptation: Stored in test substrate at test conditions over night before start of exposure
Reason for selection of species: Recommended species in the Test guidelines, its susceptibility to chemicals resembles that of true soil inhabiting species, it has a short life cycle, hatches from cocoons in 3 to 4 weeks and reaches maturity in seven to eight weeks at 20 °C

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

20 ± 2°C

pH:

pH value of the dry test substrate: 6.3 mean value of 2 single values (6.3 and 6.3, rounded to 2 significant digits)

Moisture:

Calculated water content of the test substrate at start of exposure: 30 % of dry weight
Water content of the test substrate at the end of the exposure period: 29.9 % of dry weight (single measurement)

Details on test conditions:

TEST SYSTEM
- Test container: Preserving jars, volume 1 L, covered with glass lids
- Amount of soil or substrate: 750 g ± 1 g artificial soil (wet weight)
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4 test vessels per concentration
- No. of replicates per control: 4

OTHER TEST CONDITIONS
- Photoperiod: Continuously, over the exposure period
- Light intensity: 546 Lux, mean value of 5 single values (717, 514, 483, 521, 494 [Lux]) at the start of exposure

EFFECT PARAMETERS MEASURED: Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Nominal and measured concentrations:

Nominal concentration: 0 (control), 62.5, 125, 250, 500, 1000 mg/kg soil dry weight

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

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

Duration:

14 d

Dose descriptor:

NOEC

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: none
- Any other observations: No abnormal behaviour or toxic symptoms of the worms were observed in the test substance groups or control group during the exposure.

Results with reference substance (positive control):

- Results with reference substance: The LC50 value of the reference substance for the earthworm Eisenia fetida after 14 days was not in the range of 20 mg/kg to 80 mg/kg with a value of 12.6 mg/kg. Although the LC50 is outside of the recommended range of 20-80 mg/kg the test system showed that it was sufficiently sensitive and was suitable to determine the acute toxicity of the test substance.

Validity criteria fulfilled:

yes

Description of key information

No toxic effects to Eisenia fetida could be observed at a concentration of 1000 mg/kg dw.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

The toxicity of the test substance to terrestrial organisms was investigated in a GLP guideline study according to OECD 207. In this study, no toxic effects to Eisenia fetida were observable after 14 days at the highest concentration test of 1000 mg/kg dw (BASF SE 67E0252/12G286, 2013).