Fatty acids, C16, C18 and C18-unsaturated, C12-15 alcohol (linear and branched), esters

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 Mar - 30 May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline Study

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

Qualifier:

according to guideline

Guideline:

ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction)

Qualifier:

according to guideline

Guideline:

other: Biologische Bundesanstalt für Land- und Forstwirtschaft Bundesrepublik Deutschland: Richtlinien für die Prüfung von Pflanzenschutzmitteln im Zulassungsverfahren Part VI, 2-2 (January, 1994)

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss Federal Office of Public Health, Bern, Switzerland

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): No aqueous application solution could be prepared for the dosing of the test item due to its limited water solubility. However, the test item was completely soluble in dichloromethane (organic solvent) at the required concentrations. Thus, the test item was dissolved and diluted in dichloromethane and applied to a portion of sand. An application solution for the highest test item concentration was prepared by dissolving 7500 mg of the test item completely in dichloromethane. The volume of the application solution was made up to 30 mL with dichloromethane resulting in a test item concentration of 250 mg/mL dichloromethane. Aliquots of this application solution were further diluted with dichloromethane to obtain the application solutions for the lower test concentrations. For each test concentration, 10 mL of the corresponding application solution was applied to 50 g sand in a mortar. The solvent was completely evaporated at room temperature under a hood in 1.5 hours. The remainder (sand/test item mixture) was intensively mixed by means of a pestle and then mixed on a roller mixer for 30 minutes. Then, the sand/test item mixture was divided into aliquots of 10 g (corrected for the amount of test item applied to 10 g of sand) and filled into separate vials. The individual mixtures were prepared just before test start. The 10 g aliquots of the sand/test item mixture were quantitatively incorporated into the four replicates of artificial soil per test concentration and the moisture content was adjusted.
- Controls: yes, the same artificial soil substrate with addition of sand treated analogously to the test substrates, but without test item.
- Chemical name of vehicle: dichloromethane
- Evaporation of vehicle before use: yes

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: The worms were obtained from a synchronized culture maintained at Harlan Laboratories. During breeding and keeping, they were fed with suitable food (i.e., horse manure and potatoes).
- Age at test initiation: 2-3 months old adults with a clitellum. The age of the worms from the synchronized culture did not differ by more than 4 weeks.
- Weight at test initiation: 314 - 596 mg

ACCLIMATION
- Acclimation period: One day prior to the test start, the test organisms were acclimatized to the artificial soil and test temperature.
- Acclimation conditions (same as test or not): same as test

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Test temperature:

18 - 22 °C

pH:

5.8 - 6.3 (start of the test)
5.7 - 6.0 (end of the test)

Moisture:

33-34% (start of the test)
39-42% (end of the test)

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Plastic boxes (length about 16.5 cm, width about 12 cm, height about 6.5 cm, area about 200 cm²). The test vessels were covered by transparent lids to prevent worms from escaping and to reduce evaporation during the test period. The lids were perforated to allow air exchange.
- Amount of soil or substrate: Each test vessel contained a defined amount of dry artificial soil, corresponding to a layer depth of approximately 5-6 cm
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): Sphagnum peat: 10%; Kaolin clay: 20%; Sand: 69%; CaCO3: about 0.6%; food: 1%
- Maximum water holding capacity: 64.2 g water per 100 g dry soil (ISO guideline 11268-2)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 560-720 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality and adverse effects (after 4 weeks), body weight (beginning and end of first 4 weeks), reproduction (after 8 weeks), assessment of food consumption (each feeding date)

VEHICLE CONTROL PERFORMED: no

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: A range-finding study was performed in order to derive the test concentrations. No detailed information on performance of range-finding study are given in the report.

Nominal and measured concentrations:

nominal: control, 62.5, 125, 250, 500 and 1000 mg/kg soil dw

Reference substance (positive control):

yes

Remarks:

Derosal (R)

Duration:

8 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

8 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

4 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

4 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

4 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

4 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Details on results:

- Mortality at end of exposure period: 0%
- Total mass of adults at beginning of test: 424 - 469 mg
- Changes in body weigth of live adults (% of initial weight) at end of exposure period: The mean body weight of the adult worms in the control had increased during the 4-week exposure period on average by 34 mg per worm (8% of the mean initial weight). At all test concentrations of 2-octyldodecyl isooctadecanoate, the mean increase in body weight was between 6% and 12% and statistically not significantly lower than in the control (results of a Dunnett t-test, one-sided smaller, alpha = 0.05).
- No. of offspring produced: 143 - 173 (mean)

Results with reference substance (positive control):

At the single concentration of the reference item Derosal (R) (8.33 mg formulation /kg dry soil, i.e. 5 mg a.i./kg dry soil), the mortality rate of the adult worms was 2.5%. The mean decrease of 23% in body wet weight of the earthworms in the positive control during the exposure period was significantly different to the control (results of a Student-t-test, one-sided smaller, alpha = 0.05). The reproduction rate of Eisenia fetida in the positive control was completely inhibited.

Reported statistics and error estimates:

Reproduction of earthworms: Welch t-test, one-sided smaller, alpha = 0.05
Mean body weight: Mean value not statistically significantly smaller compared to the control (Dunnett t-test, one-sided smaller, alpha = 0.05); Mean value statistically significantly smaller than the control (Student t-test, one-sided smaller, alpha = 0.05).

Table 1: Summary of effects on earthworms

Parameters	Control	2-octyldodecyl isooctadecanoate [mg/kg dry soil]					Derosal^® [mg/kg dry soil]
		62.5	125	250	500	1000	8.33
Mortality [%] after a 4-week exposure	1.25	0	0	0	0	0	2.5
Mean change in body fresh weight [%] after a 4-week exposure	8	9	6	12	8	10	-23
Mean reproduction rate (juveniles per replicate)	143	144	173	142	147	149	0.0
Reproduction rate (% of control)	100	101	121	99	103	104	0.0

Table 2: Reproduction of earthworms

Treatment [mg/kg dry soil]	Vessel No.	Surviving adults	Juvenile worms per vessel					Juvenile worms per surviving adult					Statistical evaluation
			#	mean	SD	CV [%]	% of control	#	mean	SD	CV [%]	% of control
Control	1	10	115					11.5
	2	10	101					10.1
	3	10	112					11.2
	4	10	161					16.1
	5	10	162					16.2
	6	9	140					15.6
	7	10	176					17.6
	8	10	175	143	30.0	21.0	100	17.5	14.5	3.0	21.0	100	---
62.5	1	10	113					11.3
	2	10	155					15.5
	3	10	164					16.4
	4	10	145	144	22.2	15.4	101	14.5	14.4	2.2	15.4	100	n.s.
125	1	10	178					17.8
	2	10	128					12.8
	3	10	198					19.8
	4	10	187	173	30.9	17.9	121	18.7	17.3	3.1	17.9	119	n.s.
250	1	10	169					16.9
	2	10	143					14.3
	3	10	136					13.6
	4	10	120	142	20.4	14.4	99	12.0	14.2	2.0	14.4	98	n.s.
500	1	10	89					8.9
	2	10	115					11.5
	3	10	249					24.9
	4	10	135	147	70.6	48.0	103	13.5	14.7	7.1	48.0	102	n.s.
1000	1	10	146					14.6
	2	10	120					12.0
	3	10	191					19.1
	4	10	138	149	30.2	20.3	104	13.8	14.9	3.0	20.3	103	n.s.
Derosal (R) 8.33 mg/kg dry soil	1	9	0					0.0
	2	10	0					0.0
	3	10	0					0.0
	4	10	0	0	0.0	n.a.	0.0	0.0	0.0	0.0	n.a.	0.0	---

Reference 2

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Please refer to the analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Duration:

8 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Duration:

8 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Duration:

4 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Duration:

4 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Duration:

4 wk

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Duration:

4 wk

Dose descriptor:

LOEC

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks on result:

other:

Remarks:

Source: RA-A, CAS 93803-87-3, Harlan Laboratories Ltd, 2013, E. fetida, 8 wk

Description of key information

NOEC (56 d) ≥ 1000 mg/kg soil dw (OECD 222, E. fetida, reproduction); read-across

Additional information

There are no experimental studies available, in which the toxicity of Fatty acids, C16, C18 and C18-unsaturated, C12-15 alcohol (linear and branched), esters to soil macroorganisms except arthropods was investigated. Therefore, a read-across approach was pursued to compile relevant data from two structurally and chemically related source substances in order to fulfill the standard information requirements laid down in Annex X of REACH Regulation (EC) No 1907/2006. Read-across was conducted in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5.

The source substance 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) is characterized by similar fatty acid and alcohol components as the target substance and is therefore considered a suitable representative for the assessment of the toxicity of the target substance to soil macroorganisms. A detailed read-across justification is provided in IUCLID section 13.

 

The available study with the source substance 2-octyldodecyl isooctadecanoate (CAS 93803-87-3) was conducted according to OECD guideline 222 and in compliance with GLP.

The earthworm Eisenia fetida was exposed to five (nominal) test item concentrations ranging from 62.5 – 1000 mg/kg artificial soil (dw) for 56 d. Mortality/growth was assessed after 4 weeks and reproduction was assessed after 8 weeks.

At the end of the test, no significant effects on mortality/growth and reproduction were found, resulting in a NOEC (56 d) ≥ 1000 mg/kg soil dw.

Based on the high degree of structural and chemical similarity between the target and source substances, the target substance is expected to exhibit a similar ecotoxicological profile as the source substance. Therefore, it can be concluded that the target substance Fatty acids, C16, C18 and C18-unsaturated, C12-15 alcohol (linear and branched), esters does not cause toxic effects to soil macroorganisms.