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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

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Administrative data

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
acceptable for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report date:
1993

Materials and methods

Test guidelineopen allclose all
Qualifier:
no guideline followed
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Version / remarks:
the test was performed similar to OECD TG 207 Earthworm, Acute Toxicity Tests, but extending exposure time to 21 d and including the examination of additional parameters.
Deviations:
yes
Remarks:
exposure time, parameters examined
Principles of method if other than guideline:
Artificial soil test for chronic toxicity on Eisenia fetida with an exposure time of 21 d and scoring cocoons and number of offspring in addition to mortality of adults.
Note: at the time of the study, there was no test guideline available for chronic toxicity testing in soil with Eisenia fetida.
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Phenanthrene
EC Number:
201-581-5
EC Name:
Phenanthrene
Cas Number:
85-01-8
Molecular formula:
C14H10
IUPAC Name:
phenanthrene
Test material form:
solid
Details on test material:
- Substance type: organic
- for additional information see study record
Specific details on test material used for the study:
- Name of test material (as cited in study report): phenanthrene
- Source of test material: Aldrich Chemie
- Analyticl purity: 98 %

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
SOIL
- Concentrations: 10, 100, and 1000 mg/kg soil dw
- Sampling interval: at day 0 and 21

WORM TISSUE
- Concentrations: 10, 100, and 1000 mg/kg soil dw
- Sampling interval: at day 0, 3, 7, 14, and 21

Test substrate

Vehicle:
no
Details on preparation and application of test substrate:
Preparation of the test substance:
- Method of mixing into sand: the test substance for each test concentration was dissolved in acetone and coated onto quartz sand (M32) as carrier. 350 g sand were coated for each test concentration.
- Method of mixing the coated sand into soil: 350 g portions of the coated sand were mixed into artificial soil consisting of the remainder of M32 sand (medium grain size approx. 250 µm, no organic matter), kaolin, milled sphagnum peat (7:2:1). CaCO3 was added at a ratio of 4 g/kg of dry soil. Finally the soil was wetted to 52 - 55 % humidity by taking regard of the inherent moisture of the peat used. After addition of the test substance, the spiked soil was incubated at 4 °C for 33 days to allow equilibrium partitioning of test material in the test soil

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
annelids

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
21 d

Test conditions

Test temperature:
Room temperature
pH:
6.7 - 7.0
Moisture:
Approx. 52 – 55 % (medium for adult)
Approx. 30 – 35 % (medium for cocoons)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 2 L Kilner jar
- Amount of soil or substrate: 0.75 kg/container
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: no vehicle

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10 % peat,
20 % kaolin,
70 % quartz sand,
approx. 0.4 % calcium carbonate,
10 g dried cow manure per kg soil (dw) as food for the worms
- Organic carbon (%): no data
- Maximum water holding capacity (in % dry weigth): no data
- Pretreatment of soil - main test: after addition of the test substance, the spiked soil was incubated at 4 °C for 33 days to allow equilibrium partitioning of test material in the test soil
- Pretreatment of soil - cocoon test: for the maintenance and incubation of the cocoons, the peat was sieved to a finer degree than used in acute testing (grain size 0.5 instead of 1.0 mm). The moisture content of this medium was 30-35 % instead of 52-55 %
- Storage (condition, duration): Preparation with test material one month in advance, storage at 4 °C to allow equilibrium partitioning between soil particles, organic material and soil moisture.
- Stability and homogeneity of test material in the medium: little or no degradation during storage

OTHER TEST CONDITIONS
- Photoperiod: no data
- Light intensity: 0-60 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, reproduction (cocoon production, young per cocoon) after 21 d

VEHICLE CONTROL PERFORMED: no (not applicable)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 3
- Range finding study: no data
Nominal and measured concentrations:
Nominal: 0, 10, 32, 100, 320, 1000 mg/kg (dw) artificial soil
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
ca. 5.8 mg/kg soil dw
Nominal / measured:
estimated
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
number of cocoons
Remarks on result:
other: original value (17 mg/kg soil dw) recalculated to a standard soil with 3.4 % organic matter
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
ca. 17 mg/kg soil dw
Nominal / measured:
estimated
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
number of cocoons
Remarks on result:
other: estimation based on geom. mean of measured concentrations at nominal 100 mg/kg soil dw
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
number of cocoons
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
ca. 240 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Remarks:
number of cocoons
Remarks on result:
other: EC50 calculated by maximum likelyhood model
Duration:
21 d
Dose descriptor:
LC0
Effect conc.:
320 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adult
Duration:
21 d
Dose descriptor:
LC50
Effect conc.:
> 320 - < 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adult
Duration:
21 d
Dose descriptor:
LC100
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adult
Remarks on result:
other: mortality was 50 % (20 of 40 test organisms died) at 1000 mg/kg soil dw (highest concentration tested)
Details on results:
- High decrease in test material concentrations from day 0 to day 21 (for 100 mg/kg soil dw nominal concentration: measured concentrations: 120 and 2.43 mg/kg soil dw at day 0 and day 21, respectively)
- Other biological observations: At 320 mg/kg soil (dw) and higher, adult animals were sluggish.
Reported statistics and error estimates:
No statistical analyses of the effect data

Any other information on results incl. tables

Measured phenanthrene concentrations in soil

Time of incubation

[Days]

Nominal concentration

[mg/kg soil]

0

10

100

1000

Residual concentrations in soil

0

0.025

12

120

1210

21

0.07

 1.43

2.43

   73

The pristine test substance disappeared rapidly from soil.

Measured concentrations in Eisenia fetida

Time of incubation

Nominal concentration [mg/kg soil]

[Days]

0

10

100

1000

Concentration in tissue [mg/kg dw Eisenia fetida]

0

0.03

 --

 --

  --

3

 5.0

120

 740

7

 2.2

  9.3

 120

14

 0.4

  0.47

    6.5

21

0.08

 0.06

  0.27

    6.3

Mortality and reproduction data

Test Substance

Adult

Cocoons

Concentration
(nominal/
measured)1

[mg/kg soil]

Survival after 21 d

[number survivors/

initial number]

Average number of cocoons/surviving adults

Unfertilised cocoons

Average number of

worms per fertilised cocoon

            0

39/40

56/39

6

3.7

10 (n)

40/40

56/40

2

3.6

32 (n)

40/40

51/40

5

3.7

100 (n)

40/40

51/40

3

3.5

320 (n)

40/40

17/40

3

4.3

1000 (n)

20/40

4/20

1

1.1

1TS concentrations were nominal (n)

Due to the short residence time of phenanthrene in soil, the actual exposure concentrations were lower, and therefore, the real effect concentration could be much lower, with an EC50 in the order of 10 -100 mg/kg and a NOEC for reproduction of 4 – 50 mg/kg.

Thus, reasonable estimates arrive at a NOEC (reprod.) of 17 mg/kg and EC50 of 60 mg/kg dw.

           

Applicant's summary and conclusion