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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Administrative data

toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
other information
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test system is well described and controls show it functions well.

Data source

Reference Type:
Effect of aromatic compounds, humic acids and lignins on growth of the earthworm Eisenia foetida
Roy Hartenstein
Bibliographic source:
Soil biology and biochemistry 14, pp.595-599

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
Test substance was added in varying concentrations to sludge. Sludge was placed on silt loam in a petri dish and worms were added. Survival and growth were checked over 6 weeks.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Test material form:
other: liquid
Details on test material:
Chemicals obtained from Aldrich Chemical Company, Milwaukee, Wisconsin, used without further purification

Sampling and analysis

Analytical monitoring:

Test substrate

Details on preparation and application of test substrate:
- Method of application to soil surface: Test substances were processed for assay by mixing the required amount into 100 g sludge which contained 11-15% solids. Assuming exactly 13% solids in sludge the concentrations were 0.1 to 8% (w/w) dry weight.
About 30 g spiked sludge (average 13% solids) was placed over a 4 mm depth of silt loam in a 20x100 mm petri dish.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
Stock cultures maintained on horse manure or activated sludge. 2 hatchlings each under 10 mg live weight were added to Petri dishes.

Study design

Study type:
laboratory study
Substrate type:
other: silt loam
Limit test:
Total exposure duration:
6 wk
Measurements after 2, 4 and 6 weeks
Post exposure observation period:
No post-exposure period

Test conditions

Test temperature:
24 +/- 1 degrees Celsius
no data
No exact data. Sludge contained 11 to 15% of solids, placed on silt loam layer.
Details on test conditions:
Growth was determined at 2, 4 and 6 weeks by rinsing the worms in distilled water, blotting and weighing. Worms were transferred to freshly prepared substance after 4 weeks.
Nominal and measured concentrations:
Substances tested at 0, 0.1, 1, 4 and 8 % (w/w) dry weight,
5 replicates each
Reference substance (positive control):
not specified

Results and discussion

Effect concentrationsopen allclose all
6 wk
Dose descriptor:
Effect conc.:
0.1 other: % (w/w)
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
6 wk
Dose descriptor:
other: significant growth reduction
Effect conc.:
< 0.1 other: % (w/w)
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
After 6 weeks at 0.1 % mortality was 100%. This was the lowest test concentration
Details on results:
Hatchlings in control dishes would gain more than 500 mg in 6 weeks. An average weight of 582 mg +/- 85 SD, with a c.v. of 14.6% was attained by controls in 6 weeks in 45 similar assays during 16 months. This reflects both variation in earthworm growth and sludge quality. Mortality in controls was less than 2%.
Dimethoxybenzenes were severely more toxic than di- and trihydroxybenzenes and monosubstituted hydroxybenzenes
Results with reference substance (positive control):
no data
Reported statistics and error estimates:
Significant differences for growth reduction were detected by the Neuman-Keul test (Zar 1974) (but not relevant for this test substance due to 100 % mortality). Mortality was said to be significant if 5 or more earthworms died.

Applicant's summary and conclusion

Validity criteria fulfilled:
not applicable
mortality in control very low, and high growth make test reliable
100% mortality of the test organism Eisenia fetida was observed at the lowest test concentration of 0.1 % (w/w in sludge) in a mixture of active sludge and soil.
Executive summary:

Eisenia fetida was tested in spiked activated sludge with concentrations from 0.1 to 8% (w/w) dry wt, including controls. The test duration was 6 weeks. Blank control performance of the test was good. For the test substance, 100 % mortality was observed at the lowest test concentration of 0.1 % (w/w in sludge dry weight) in a mixture of activated sludge and soil.