Perhydroimidazo[4,5-d]imidazole-2,5-dione

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:

2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test substance: Acetylendiharnstoff.
Batch number: Fixapret 140 Nr. 766.
Purity: 96.3g/100g.
Impurities: urea, sulfuric acid, glyoxal <0.1%.

Analytical monitoring:

yes

Vehicle:

yes

Details on preparation and application of test substrate:

Test substrate
69.6% Quartz sand
20.0% Kaolin clay
10.0% Sphagnum peat
0.5% Calcium carbonate

Preliminary investigations of test substrate
The water content of the test substrate before the addition of liquid was determined by drying a part of the dry test substrate at 105°C (24hrs) and following reweighting. Water content was disregarded to the calculation of the water content in the test.

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

Test species: Eisenia foetida.
Supplier: Regenwurmfarm Tacke GmbH, Klosterdiek 61, 46325 Borken-Burlo, from 17 Jan 2002.
Weight of one animal: Mean value 339mg at start of the exposition (mean value of all test concentrations and control subjects).
Age of animals at the start of the test: Between 2-12 month(s).
Feeding: None.
Acclimatisation: 24hrs before the start of the exposition at test conditions without test substance.

Study type:

laboratory study

Substrate type:

artificial soil

Total exposure duration:

14 d

Test temperature:

Exposition phase: 21-23°C.
Adaption phase: 21-23°C.
Measured continously during the adaptation and exposition phase in the exhaust of test room.

pH:

pH of test substrate: 6 ± 0.5.
Measured before and beginning of the adaptation according to DIN/ISO 10 390.

Moisture:

Water content of the test substrate in the exposition phase: 33g/100g dry weight ± 2g/100g.
Water content of the test substrate in adaptation phase: 33g/100g dry weight ± 2g/100g.
Water constant of the dry test substrate: <2g/100g dry weight.
Measurement of water content of the moistened test substrate: At the start and end of the exposition (at the end of the test exposition in a separate vessel close to the test vessels without test substance).
Measurement of water content of the dry test substrate: Before start of adaptation.

Details on test conditions:

Adaptation phase: 1 day.
Duration of exposition phase: 14 days.
Light intensity in exposition phase: 400-800 Lux.
Light intensity in adaptation phase: 400-800 Lux.

Measurement of light intensity: One time at the start of the adaptation and one time at the start of the exposition phase.
Measurement of the weight of the earthworms: origin and end of the exposition.

Test vessel: 1L glass jars with glass lids.
Test volume 750g artifical soil (moist weight).
Number of animals/vessel: 10.
Total number of animals/concentration: 40.
Number of concentrations (without control): 5.
Number of replicates: 4.
Number of parallels without animals: One for determination of water content.

Application of test substance
Test substance was weighed into 10g of quartz sand and mixed well. Afterwards, these were mixed with 740g of the dry test substrate and 250mL of demineralised water (added to adjust the water content at 33± 2% of the dry weight. The same was carried out for the control but without the test substance.

Nominal and measured concentrations:

Nominal concentrations (mg/kg): 0 (control), 62.5, 125, 250, 500, 1000.
Addition of test substance (respective to concentrations above) (mg): 0, 47, 93.6, 187.5, 374.8, 750.

Reference substance (positive control):

yes

Remarks:

2-chloracetamide

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

>= 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

other: EC80

Effect conc.:

> 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

other: LC10

Effect conc.:

>= 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

LC100

Effect conc.:

> 1 000 mg/kg soil ww

Nominal / measured:

nominal

Basis for effect:

mortality

Results with reference substance (positive control):

The results from the positive control was utilised for the validity criteria of the test. The mortality in the controls did not exceed 10% at the end of the test. The LC50 (14 days) of the control substance 2-chloracetamide was 16.5 mg/kg.

Reported statistics and error estimates:

There was no statistical calculation.

Validity criteria fulfilled:

yes

Conclusions:

The 14 day study showed no signs of toxicity (LC50) for glycoluril in the earthworm at the tested concentration (1,000 mg/kg) it was therefore concluded the LC50 is above 1,000mg/kg.

Executive summary:

To determine the acute toxicity of the test substance in earthworms (Eisenia foetida) the OECD 207 (April 1984) was followed, the tests were carried out in a laboratory with a GLP Certificate. A total of fourty (40) earthworms were tested per concentration (0, 62.5, 125, 250, 500 and 1,000 mg/kg). Mortality was assessed seven (7) and fourteen (14) days after application. At the end of the test period, no negative impact on worm biomass was detected (<10% inhibition). Additionally, no other particular behavioural or morphological changes were observed.

 

It was therefore concluded that the test substance is non-toxic to earthworms; LC50 >1,000 mg/kg based on analytically confirmed nominal concentrations and therefore does not meet the classification criteria under Regulation (EC) No. 1272/2008.

Description of key information

To determine the acute toxicity of the test substance in earthworms (Eisenia foetida) the OECD 207 (April 1984) was followed, the tests were carried out in a laboratory with a GLP Certificate. A total of fourty (40) earthworms were tested per concentration (0, 62.5, 125, 250, 500 and 1,000 mg/kg). Mortality was assessed seven (7) and fourteen (14) days after application. At the end of the test period, no negative impact on worm biomass was detected (<10% inhibition). Additionally, no other particular behavioural or morphological changes were observed.

 

It was therefore concluded that the test substance is non-toxic to earthworms; LC50 > 1,000 mg/kg based on analytically confirmed nominal concentrations and therefore does not meet the classification criteria under Regulation (EC) No. 1272/2008.

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:

1 000 mg/kg soil dw