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Toxicity to soil macroorganisms except arthropods

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Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Sep - Nov 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP guideline study tested with the source substance CAS 25151-96-6. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
GLP compliance:
yes (incl. QA statement)
Remarks:
Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, date of inspection 14 September 2010
Analytical monitoring:
no
Vehicle:
yes
Details on preparation and application of test substrate:
- Method of mixing into soil: the test substance was dissolved in acetone. The solution was applied to 10 g quartz sand. After evaporation of the solvent the sand/substance mixture was mixed with the artifical soil.
- Controls: the solvent was added to 10 g quartz sand. After evaporation the sand was mixed with the artificial soil
- Chemical name of vehicle: acetone
- Evaporation of vehicle before use: yes
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: Regenwurmfarm Tacke, Borken, Germany. Specimens used in the test were bred in the laboratory of the Fraunhofer IME.
- Age at test initiation: 2 - 12 month
- Weight at test initiation: 250 - 600 mg (ww)

ACCLIMATION
- Acclimation period: 5 days
- Acclimation conditions: same as in test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
20 ± 2 °C
pH:
6.4 – 7.0
Moisture:
54.7 – 60.0% of WHC
Details on test conditions:
TEST SYSTEM
- Test container (material, size): plastic containers, 17 x 12 x 6 cm
- Amount of soil or substrate: 500 - 650 g dry mass
- 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: 4

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition: 10% Sphagnum peat, 20% Kaolinite, 70% Industrial quartz sand


OTHER TEST CONDITIONS
- Photoperiod: 16:8 h day-night regime
- Light intensity: 682 – 728 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and biomass of surviving adult earthworms after 28 days and number of living offspring after 56 days

VEHICLE CONTROL PERFORMED: yes

Nominal and measured concentrations:
500 and 1000 mg/kg soil (dm)
Reference substance (positive control):
yes
Remarks:
Carbendazim
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
- Mortality at end of exposure period: none
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: -1.9% at 500 mg/kg and 1.1% at 1000 mg/kg
- No. of offspring produced: 219 specimen at 500 mg/kg and 226 specimen at 1000 mg/kg
- Morphological abnormalities: none
- Behavioural abnormalities: none
- Other biological observations: none
Results with reference substance (positive control):
NOEC (mortality) ≥ 3 mg/kg
NOEC (reproduction): 0.75 mg/kg; EC50: 1.7 mg/kg
Tests with Carbendazim are performed once a year
Reported statistics and error estimates:
Statistic calculations were performed with ToxRat® Pro 2.10, a statistical software package for ecotoxicity response analysis by ToxRat® Solutions.

The weight and survival rate of adult earthworms was determined after 28 days of exposure. The hatched offspring was collected after a test period of 56 days.

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 Nov 2012 - 10 Jan 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP Guideline study tested with the source substance CAS 68604-44-4. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
GLP compliance:
yes (incl. QA statement)
Remarks:
Staatsministerium für Umwelt und Landwirtschaft, Freistaat Sachsen, date of inspection 16-17 June 2010 and 28 September 2010
Analytical monitoring:
no
Vehicle:
yes
Details on preparation and application of test substrate:
- Method of mixing into soil: an exact weighed amount of the test substance was mixed with finely quartz sand (as vehicle) by using a pestle and mortar. For the stock mixture, 5 g of the test substance was mixed with 95 g of quartz sand. The stock mixture of the reference substance was diluted to the exact amount for each test vessel. The test substance with quartz sand and the reference substance with quartz sand were mixed thoroughly by intensive stirring with the artificial soil using the mixer with bowl.
Test organisms (species):
other: Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: Eurofins Agrosciences Services EcoChem GmbH, Niefern, Germany
- Age at test initiation: 12 month (with clitellum)
- Weight at test initiation: 285 - 584 mg (ww)

ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions: same as in test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
56 d
Test temperature:
20 ± 2 °C
pH:
6.0 ± 0.5
Moisture:
32 g/100 g dw (at test start)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass with glass lids
- Amount of soil or substrate: 500 g dry mass
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 8
- No. of replicates per vehicle control: 8
- No. of replicates per positive control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition: 10% sphagnum peat, 20% kaolin clay, 70% industrial quartz sand

OTHER TEST CONDITIONS
- Photoperiod: 16:8 h day-night regime
- Light intensity: 778 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and biomass of surviving adult earthworms after 28 days and number of living offspring after 56 days

VEHICLE CONTROL PERFORMED: yes
Nominal and measured concentrations:
nominal: 1000 mg/kg dw soil
Reference substance (positive control):
yes
Remarks:
Benomyl
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
- Mortality at end of exposure period: none
- Changes in body weigth of live adults at end of exposure period: no significant changes at the test concentration of 1000 mg/kg dw
- No. of offspring produced: 117 juveniles in the vehicle control and 123 at 1000 mg/kg dw
- Morphological abnormalities: none
- Behavioural abnormalities: none
- Other biological observations: none
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: The positive control with 1.25 mg/kg dw soil Benomyl showed 57.6% effect on reproduction
Reported statistics and error estimates:
The statistical calculation was carried out using the statistical program SigmaPlot 11.0.

Mortality, number of hatched juveniles, inhibition of reproduction and biomass change

Treatment (mg/kg dry weight)

Mean mortality after 4 weeks (%)

Juveniles per vessel (mean)

Mean inhibition of reproduction (%)

Mean biomass change after 4 weeks (%)

Control

0

117

-

35.6

1000

0

123

-5.1

39.2

Reference substance

0

50

57.6

44.1

(negative value: increase of reproduction)

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 - 19 Nov 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study with acceptable restrictions (no purity, no reference substance)
Qualifier:
according to guideline
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations:
- Sampling method: Samples were collected on Day 2 of dry mixing, Day 6 of hydration equilibration, Day 0, prior to the addition of worms for the control and treatment and at test termination (Day 14). The samples were collected using a stainless steel spatula and weighed into 40 mL VOA vials. Three replicates were collected from the treatment soil and duplicate from the control. The samples collected at termination were from a pooled sample where all four replicates for each treatment group were composited.
- Sample storage conditions before analysis:
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): The treated artificial soil was prepared by spiking 300 grams of peat with the appropriate amount of test material using volumetric pipettes and/or stainless steel and glass syringe and mixed prior to the addition of clay and sand. The treated soil and control soil was mixed on a roller mill for three days. After hydration with distilled water the soils were placed in an environment chamber at test temperature to equilibrate for 8 days.
- Controls: A control soil was prepared without the test substance.
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: Earthworm
- Source: Lot W-43, Carolina Biological Supply Co. 2700 York Road, Burlington, NC 27215-3398
- Age at test initiation (mean and range, SD): Worms with clitellum.
- Weight at test initiation: 300 - 600 mg

ACCLIMATION
- Acclimation period: 12 October - 05 November 2012
- Acclimation conditions (same as test or not): Worms were held and evaluated in worm bedding moistened with distilled water from 03 October 2012 until 12 October 2012. All study worms were transferred from worm bedding and conditioned in an artificial soil bed from 12 October 2012 until study initiation on 05 November 2012.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
14 d
Test temperature:
19.87°C -20.16°C
pH:
Control: 6.43 (Day 0); 6.41 (Day 14)
1000 mg/kg: 6.31 (Day 0); 6.43 (Day 14)
Moisture:
Control: 27.4 - 30.6% (Day 0)
1000 mg/kg: 24.2 - 29.9% (Day 0)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1 L glass jars
- Amount of soil or substrate: 750 g of soil
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- Other: Each chamber was covered with perforated plastic film during the study to maintain hydration and allow some air circulation.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 70% sand, 20% kaolin clay, 10% peat (according to OECD 207)
- Other: Distilled water- UV sterilized, deionized well water is distributed throughout the testing facikity via PVC stainless steel pipes.

OTHER TEST CONDITIONS
- Photoperiod: continuous
- Light intensity: 719 - 728 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Observations for mortality were performed on Days 7 and 14 of the test. Each test chamber was weighed after the addition of hydrated soil and worms on Day 0 to obtain a starting weight for each test chamber. On Day 7 and 14, the chambers were re-weighed to monitor any water weight loss. All surviving worms were pooled per replicate and weighed at the end of the test.
Nominal and measured concentrations:
nominal: 1000 mg/kg dw
measured:
991 mg/kg (average initial concentration prior to soil hydration)
205 mg/kg (final analysis of the soil concentration)
see table 1 under "Any other information on materials and methods" for detailed information
Key result
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: No mortality was observed at test termination in both the control and treatment group.
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: The weight indicate that there were no sublethal effects observed.

Table 2: Worm Percent Survival and Worm Weights

Treatment Group (mg/kg)

Percent Worm Survival

Mean Wet Worm Weight(g)

Day 7*

Day 14*

Day 0**

Day 14**

Control

 

 

 

 

Replicate1

10/10 (100%)

10/10 (100%)

4.2498

3.1220

Replicate2

10/10 (100%)

10/10 (100%)

4.7216

3.6122

Replicate3

10/10 (100%)

10/10 (100%)

4.3836

3.3593

Replicate4

10/10 (100%)

10/10 (100%)

4.9141

4.1418

1000 mg/kg

 

 

 

 

Replicate1

10/10 (100%)

10/10 (100%)

4.7145

3.3925

Replicate2

10/10 (100%)

10/10 (100%)

4.4095

3.5429

Replicate3

10/10 (100%)

10/10 (100%)

4.6580

3.6534

Replicate4

10/10 (100%)

10/10 (100%)

4.5076

3.5073

*Worms were observed, counted and placed back in original soil and chamber
**Worms were carefully rinsed with de-ionized water, bottled dry and weighed by replicate. The mean wet weight was determined an average of the replicate weights for each group.

Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25 Oct 2012 - 20 Feb 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP - Guideline study, tested with the source substance 2-ethyl-2-[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate (CAS 78-16-0). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Qualifier:
equivalent or similar to guideline
Guideline:
ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Vehicle:
yes
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): A stock solution was prepared by dissolving the test substance in aceton (250 mg test substance/mL acetone). A serial dilution of the stock solution was prepared for the required test substance concentrations. The solutions were subsequently mixed with sand. The mixture was added to 496.4 g ww of artificial soil. 2.8 g calcium carbonate, 5 g food (horse manure), and 143.6 mL purified water were added and the substate was mixed in a laboratory mixer.
- Chemical name of vehicle: acetone
- Evaporation of vehicle before use: yes
Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: Blades Biological Ltd, Cowden, United Kingdom
- Age at test initiation: adult, with clitellum
- Weight at test initiation (range): 310 - 594 mg

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions : same as in test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Remarks:
After 28 days the adult earthworms were removed and the cocoons and juveniles remained in the test substrate for another 28 days
Test temperature:
19 - 22 °C
pH:
6.1 - 6.3
Moisture:
33-41% of water holding capacity (WHC: 64.2 g /100 g dry soil).
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Plastic boxes (16.5 cx 12 x 6.5 cm, area about 200 cm2)
- Amount of soil or substrate: 650 g
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8 (incl. vehicle control)

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): 10% Sphagnum peat, 20% Kaolin clay, 69% Sand


OTHER TEST CONDITIONS
- Photoperiod: 16:8 h day-night regime
- Light intensity: 656-749 Lux

EFFECT PARAMETERS MEASURED:
Mortality and body weight of adult earthworms after 28 days, reproduction rate after 56 days



Nominal and measured concentrations:
Nominal test substance concentrations: 62.5, 125, 250, 500 and 1000 mg/kg dry soil.
Reference substance (positive control):
yes
Remarks:
Carbendazim, Batch No.: 12001/03
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
- Mortality at end of exposure period: control: no mortality observed; test substance: <2.5%
Results with reference substance (positive control):
75% mortality at a reference substance concentration of 8.33 mg/kg dry soil
Reported statistics and error estimates:
Changes in body weight and differences regarding mortalilty between treatment and control were evaluated by multiple Dunnett t-test (Dunnett, 1955; Dunnett, 1964) after a one-way analysis of variance (ANOVA).
The body weight change of the surviving worms in the positive control was compared to the control and was statistically evaluated by means of a Student-t test (Sachs, 1984).
Reproduction rates were analysed using a multiple Dunnett t-test [Dunnett, 1955; Dunnett, 1964] after a one-way analysis of variance (ANOVA).

Mortality, number of hatched juveniles and changes in body weight

Treatment (mg/kg dry weight)

Mortality (%)

after a 4-week exposure

Juveniles per vessel (mean)

Juvenils per vessel (% of control)

Mean body weight

per worm (% difference from test start)

Control

0

168

100

9

62.5

0

137

81

17

125

0

141

84

14

250

2.5

150

90

15

500

2.5

155

92

17

1000

0

152

90

14

Reference substance

7.5

0

0

-54
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 Nov 2012 - 10 Jan 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP - Guideline study, tested with the source substance Fatty acids, C16-18 and C18-unsatd., esters with pentaerythritol (CAS 85711-45-1). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
GLP compliance:
yes (incl. QA statement)
Remarks:
Staatsministerium für Umwelt und Landwirtschaft, Freistaat Sachsen, date of inspection 16-17 June 2010 and 28 September 2010
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil: an exact weighed amount of the test substance was mixed with finely quartz sand by using a pestle and mortar. For the stock mixture, 5 g of the test substance was mixed with 95 g of quartz sand. The stock mixture of the reference substance was diluted to the exact amount for each test vessel. The test substance with quartz sand and the reference substance with quartz sand were mixed thoroughly by intensive stirring with the artificial soil using the mixer with bowl. The water content was then adjusted at 32 g/100 g dw (with deionised water).
- Controls: yes, artificial soil without test substance
Test organisms (species):
other: Eisenia andrei
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Source: Eurofins Agrosciences Services EcoChem GmbH, Niefern, Germany
- Age at test initiation: 12 month (with clitellum)
- Weight at test initiation: 285 - 584 mg (ww)

ACCLIMATION
- Acclimation period: 2 days
- Acclimation conditions: same as in test
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
56 d
Test temperature:
20 ± 2 °C
pH:
6.0 ± 0.5
Moisture:
32 g/100 g dw (at test start)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass with glass lids
- Amount of soil or substrate: 500 g dry mass
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 8
- No. of replicates per vehicle control: 8
- No. of replicates per positive control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition: 10% sphagnum peat, 20% kaolin clay, 70% industrial quartz sand

OTHER TEST CONDITIONS
- Photoperiod: 16:8 h day-night regime
- Light intensity: 778 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and biomass of surviving adult earthworms after 28 days and number of living offspring after 56 days

VEHICLE CONTROL PERFORMED: yes
Nominal and measured concentrations:
nominal: 1000 mg/kg dw soil
Reference substance (positive control):
yes
Remarks:
Benomyl
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
- Mortality at end of exposure period: none
- Changes in body weigth of live adults at end of exposure period: no significant changes at the test concentration of 1000 mg/kg dw
- No. of offspring produced: 117 juveniles in the vehicle control and 150 at 1000 mg/kg dw.
- Morphological abnormalities: none
- Behavioural abnormalities: none
- Other biological observations: none
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: The positive control with 1.25 mg/kg dw soil Benomyl showed 57.6% effect on reproduction
Reported statistics and error estimates:
The statistical calculation was carried out using the statistical program SigmaPlot 11.0.

Mortality, number of hatched juveniles, inhibition of reproduction and biomass change

Treatment (mg/kg dry weight)

Mean mortality after 4 weeks (%)

Juveniles per vessel (mean)

Mean inhibition of reproduction (%)

Mean biomass change after 4 weeks (%)

Control

0

117

-

35.6

1000

0

150

-28.8*

42.0

Reference substance

0

50

57.6

44.1

 * In this concentration earthworms produced more juveniles than earthworms in the control group. This statistical significant effect is not a negative reproduction effect. Therefore, the NOEC for reproduction was determined as ≥ 1000 mg/kg dw soil.

Description of key information

NOEC (56 d) ≥ 1000 mg/kg dw for Eisenia fetida (OECD 222)

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

Additional information

CAS 71010-76-9

The short term toxicity of decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid was investigated according to OECD guideline 207 and under GLP (Kelley, 2013). Using artificial soil that was mixed with the test substance. Earthworms of the species Eisenia fetida were exposed to the test substance for 14 days and then removed from the test substrate. The survival rate and growth (body weight) of the specimen were determined. Neither mortality or a decrease in body weight of the adult earthworms was not observed. Thus a NOEC 1000 mg/kg soil based on mortality was reported. A NOEC ≥ 1000 mg/kg soil based on the reproduction rate was determined.

 

CAS 78-16-0

In the study with a shorter chained poloyl ester the long-term toxicity of 2-ethyl-2-[[(1-oxoheptyl)oxy]methyl]propane-1,3-diyl bisheptanoate to earthworms was investigated according to OECD guideline 222 and under GLP (Eisner, 2013). The test was conducted with artificial soil that was mixed with the test substance. Earthworms of the species Eisenia fetida were exposed to the test substance for 28 days and then removed from the test substrate. The survival rate and growth (body weight) of the specimen were determined. After another 28 days the hatched juveniles were collected from the test substrate and the reproduction rate was recorded. Mortality or a decrease in body weight of the adult earthworms was not observed. Thus a NOEC 1000 mg/kg soil based on mortality was reported. A NOEC ≥ 1000 mg/kg soil based on the reproduction rate was determined.

 

CAS 68604-44-4

In the study the chronic toxicity of fatty acids, C16-18 and C18-unsatd., tetraesters with pentaerythritol to earthworms was conducted according to OECD 222 under GLP conditions (Amsel, 2013). Eisenia andrei was exposed to a test concentration of 1000 mg/kg dw in artificial soil for 56 days. Mortality and reproduction were recorded at the end of the study. No effects were observed resulting in a NOEC (28 d) of ≥ 1000 mg/kg dw based on mortality and a NOEC (56 d) of ≥ 1000 mg/kg dw based on reproduction. Due to these results it can be concluded that fatty acids, C16-18 and C18-unsatd., tetraesters with pentaerythritol (CAS 68604-44-4) will not exhibit effects on soil macroorganisms.

 

CAS 25151-96-6

A further chronic test with substance 2,2-bis(hydroxymethyl)-1,3-propanediyl dioleate was conducted according to OECD 222 under GLP conditions (Simon, 2013). Eisenia fetida was exposed to the test concentrations 500 and 1000 mg/kg dw in artificial soil for 56 days. Mortality and reproduction were recorded at the end of the study. No effects were observed resulting in a NOEC (28 d) of ≥ 1000 mg/kg dw based on mortality and a NOEC (56 d) of ≥ 1000 mg/kg dw based on reproduction.