Fatty acids, C16-18 and C18-unsatd., esters with propylene glycol

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 Jan - 28 Mar 2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP Guideline study. In accordance 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 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 11268-1 (Effects of Pollutants on Earthworms. 1. Determination of Acute Toxicity Using Artificial Soil Substrate)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Landwirtschaft und Forsten, Wiesbaden, Germany

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil: A stock solution was prepared by weighing 5.33 g of the test substance in 13 mL acetone. The stock solution was diluted to obtain the appropriate test concentrations. 5 mL of the stock solution or the corresponding dilution were added to 20 g of fine quartz sand. After evaporation of the solvent, 2030 g of dry artificial soil were added to the test item blended sand. The soil was mixed using a laboratory mixer; in the course of applying, the soil was ventilated and moistened.
- Controls: yes, artificial soil control
- 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
- Common name: earthworm
- Source: Bred under standardised conditions (OECD 207) by IBACON, Rossdorf, Germany.
- Age at test initiation: approx. 10 months, with clitellum
- Weight at test initiation: 300 - 600 mg

ACCLIMATION
- Acclimation period: 1 d
- Acclimation conditions: same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

19 - 20 °C

pH:

5.5 - 5.9

Moisture:

test start: 27.5 - 28.2%
test end: 26.3 - 27.2%

Details on test conditions:

TEST SYSTEM
- Test container: Normal bottling jars (1 L), loosely covered by glass-lids to enable exchange of air and to minimise evaporation.
- Amount of soil or substrate: 500 g dw
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE
- Composition: according to OECD guideline 207
- Storage: Artificial soil was stored until use in a closed plastic-container.


OTHER TEST CONDITIONS
- Photoperiod: continuous light
- Light intensity: 464 - 772 lux

EFFECT PARAMETERS MEASURED: Mortality (day 7 and 14), Behavioural abnormalities (day 7 and 14), Mean body weight (day 0 and 14)

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations: 1, 10, 100, 500, 100 mg/kg dw
- Results used to determine the conditions for the definitive study: The range-finding study did not result in any mortality and no significant reduction of body weight.

Nominal and measured concentrations:

nominal: 0, 62.5, 125.0, 250.0, 500.0, 1000.0 mg/kg dw

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

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:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Details on results:

- Mortality at end of exposure period: 2.5% at a concentration of 62.5 mg/kg dw (not significant). 0% in the control.
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: 9.7% increase in body weight at 1000 mg/kg dw
- Morphological abnormalities: One worm in replicate 2 of the control group was observed with cut off body parts.

Results with reference substance (positive control):

- Relevant effect levels: LC50 (14 d) = 21.5 mg/kg dw (95% CL: 19.9 - 23.2 mg/kg dw)

Reported statistics and error estimates:

Body weight changed significantly compared to the control at a concentration of 1000 mg/kg dw (Dunn-test, α = 0.05).

Validity criteria fulfilled:

yes

Conclusions:

After 14 days the EC50 was determined to be > 1000 mg/kg soil dw.

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

05 Jan - 28 Mar 2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP Guideline study. In accordance 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.

Justification for type of information:

Please refer section 13 for read across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 11268-1 (Effects of Pollutants on Earthworms. 1. Determination of Acute Toxicity Using Artificial Soil Substrate)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Landwirtschaft und Forsten, Wiesbaden, Germany

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil: A stock solution was prepared by weighing 5.33 g of the test substance in 13 mL acetone. The stock solution was diluted to obtain the appropriate test concentrations. 5 mL of the stock solution or the corresponding dilution were added to 20 g of fine quartz sand. After evaporation of the solvent, 2030 g of dry artificial soil were added to the test item blended sand. The soil was mixed using a laboratory mixer; in the course of applying, the soil was ventilated and moistened.
- Controls: yes, artificial soil control
- 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
- Common name: earthworm
- Source: Bred under standardised conditions (OECD 207) by IBACON, Rossdorf, Germany.
- Age at test initiation: approx. 10 months, with clitellum
- Weight at test initiation: 300 - 600 mg

ACCLIMATION
- Acclimation period: 1 d
- Acclimation conditions: same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

14 d

Test temperature:

19 - 20 °C

pH:

5.5 - 5.9

Moisture:

test start: 27.5 - 28.2%
test end: 26.3 - 27.2%

Details on test conditions:

TEST SYSTEM
- Test container: Normal bottling jars (1 L), loosely covered by glass-lids to enable exchange of air and to minimise evaporation.
- Amount of soil or substrate: 500 g dw
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE
- Composition: according to OECD guideline 207
- Storage: Artificial soil was stored until use in a closed plastic-container.


OTHER TEST CONDITIONS
- Photoperiod: continuous light
- Light intensity: 464 - 772 lux

EFFECT PARAMETERS MEASURED: Mortality (day 7 and 14), Behavioural abnormalities (day 7 and 14), Mean body weight (day 0 and 14)

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations: 1, 10, 100, 500, 100 mg/kg dw
- Results used to determine the conditions for the definitive study: The range-finding study did not result in any mortality and no significant reduction of body weight.

Nominal and measured concentrations:

nominal: 0, 62.5, 125.0, 250.0, 500.0, 1000.0 mg/kg dw

Reference substance (positive control):

yes

Remarks:

2-Chloroacetamide

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:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Duration:

14 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: body weight

Details on results:

- Mortality at end of exposure period: 2.5% at a concentration of 62.5 mg/kg dw (not significant). 0% in the control.
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: 9.7% increase in body weight at 1000 mg/kg dw
- Morphological abnormalities: One worm in replicate 2 of the control group was observed with cut off body parts.

Results with reference substance (positive control):

- Relevant effect levels: LC50 (14 d) = 21.5 mg/kg dw (95% CL: 19.9 - 23.2 mg/kg dw)

Reported statistics and error estimates:

Body weight changed significantly compared to the control at a concentration of 1000 mg/kg dw (Dunn-test, α = 0.05).

Validity criteria fulfilled:

yes

Conclusions:

After 14 days the EC50 was determined to be > 1000 mg/kg soil dw.

Reference 3

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 Jun - 24 Oct 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP Guideline study. In accordance 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:

EU Method C.8 (Toxicity for Earthworms: Artificial Soil Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: The test substance was mixed-up with quartz sand. The artificial soil was prepared according to guideline and mixed with the test substance in quartz sand and water to obtain a total moisture of approx. 40% (related to the dry weight).
- Controls: yes, artificial soil without test substance

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: Eisenia GmbH & Co. Produktions- und Vertriebs-KG, Kapellenstr. 25, Wiesbaden, Germany
- Age at test initiation (mean and range, SD): at least two month old
- Weight at test initiation (mean and range, SD): 300 - 595 mg
- Other: Test species were held in slightly closed Makrolen-vessels. The substrate used for the cultivation was made from peat and cowshed manure obtained from ASEF. They were fed with fresh horse dung or dried cowshed manure if needed.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

20 ± 2 °C

pH:

6.3 - 6.4

Moisture:

36-39%

Details on test conditions:

TEST SYSTEM
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

OTHER TEST CONDITIONS
- Photoperiod: continuous light
- Light intensity: approx. 400 - 600 lux

EFFECT PARAMETERS MEASURED: Mortality was recorded on day 7 and 14.

Nominal and measured concentrations:

Nominal: control - 1000 mg/kg dw

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Key result

Duration:

14 d

Dose descriptor:

LC0

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

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

Details on results:

- Mortality at end of exposure period: 5.0% (control); 7.5% (treatment)

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: 100% mortality at 100 mg/kg dw and 0% mortality at 30 mg/kg dw after 7 days
- Other: Moisture: 38 – 39%, pH: 6.0 – 6.1

Table 1: Mortality of test organisms after day 7 and day 14 of exposure

concentration (mg/kg dw)	Mortality of test organisms
	day 7			day 14
	alive	dead	%	alive	dead	%
control	39	1	2.5	38	2	5.0
1000	39	1	2.5	37	3	7.5

Validity criteria fulfilled:

yes

Conclusions:

After 14 days the LC0 was determined to be 1000 mg/kg soil dw und the LC50 was determined to be > 1000 mg/kg soil dw.

Reference 4

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

27 Jun - 24 Oct 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

GLP Guideline study. In accordance 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.

Justification for type of information:

Please refer section 13 for the read across justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

EU Method C.8 (Toxicity for Earthworms: Artificial Soil Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: The test substance was mixed-up with quartz sand. The artificial soil was prepared according to guideline and mixed with the test substance in quartz sand and water to obtain a total moisture of approx. 40% (related to the dry weight).
- Controls: yes, artificial soil without test substance

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: Eisenia GmbH & Co. Produktions- und Vertriebs-KG, Kapellenstr. 25, Wiesbaden, Germany
- Age at test initiation (mean and range, SD): at least two month old
- Weight at test initiation (mean and range, SD): 300 - 595 mg
- Other: Test species were held in slightly closed Makrolen-vessels. The substrate used for the cultivation was made from peat and cowshed manure obtained from ASEF. They were fed with fresh horse dung or dried cowshed manure if needed.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Test temperature:

20 ± 2 °C

pH:

6.3 - 6.4

Moisture:

36-39%

Details on test conditions:

TEST SYSTEM
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4

OTHER TEST CONDITIONS
- Photoperiod: continuous light
- Light intensity: approx. 400 - 600 lux

EFFECT PARAMETERS MEASURED: Mortality was recorded on day 7 and 14.

Nominal and measured concentrations:

Nominal: control - 1000 mg/kg dw

Reference substance (positive control):

yes

Remarks:

chloroacetamide

Key result

Duration:

14 d

Dose descriptor:

LC0

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

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

Details on results:

- Mortality at end of exposure period: 5.0% (control); 7.5% (treatment)

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: 100% mortality at 100 mg/kg dw and 0% mortality at 30 mg/kg dw after 7 days
- Other: Moisture: 38 – 39%, pH: 6.0 – 6.1

Table 1: Mortality of test organisms after day 7 and day 14 of exposure

concentration (mg/kg dw)	Mortality of test organisms
	day 7			day 14
	alive	dead	%	alive	dead	%
control	39	1	2.5	38	2	5.0
1000	39	1	2.5	37	3	7.5

Validity criteria fulfilled:

yes

Conclusions:

After 14 days the LC0 was determined to be 1000 mg/kg soil dw und the LC50 was determined to be > 1000 mg/kg soil dw.

Description of key information

LC50 (14 d) > 1000 mg/kg dw for Eisenia fetida (OECD 207/EU Method C.8)

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil macroorganisms:

1 000 mg/kg soil dw

Additional information

Since no studies investigating the toxicity to soil macroorganism of the test substance are available, in accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5 a read across was conducted to the two structurally related substances butylene glycol dicaprylate / dicaprate (CAS 853947-59-8) and decanoic acid, mixed diesters with octanoic acid and propylene glycol (CAS 68583-51-7). This read-across is justified in detail in the overall summary (IUCLID chapter 6.3) and within the analogue justification in IUCLID Section 13.

The study with butylene glycol dicarprylate/dicaprate was performed according to OECD 207 under GLP conditions (Lührs, 2001). Eisenia fetida was exposed to 5 different test substance concentrations up to 1000 mg/kg dw in artificial soil for 14 d. Mortality and body weight were recorded at the end of the study. No mortality was observed resulting in a LC50 of > 1000 mg/kg dw. The body weight increased significantly at a test concentration of 1000 mg/kg dw. Therefore, a NOEC of 500 mg/kg dw was determined.

Eisenia foetida foetida was used in the second study according to EU Method C.8 (GLP) to evaluate the toxicity of decanoic acid, mixed diesters with octanoic acid and propylene glycol to soil macroorganisms (Scholz, 1996). A limit test with a concentration of 1000 mg/kg dw did not result in mortality after 14 d of exposure. Therefore, a LC50 > 1000 mg/kg dw was determined.

Based on the results from structurally related read-across substances (in accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5) which are characterized by an equal ecotoxicological profile, it can be concluded that the target substance will not exhibit toxic effects to soil macroorganisms.