Reaction product of 3,5,5-trimethyl-hexanoic acid and 2-methylpropanoic acid and pentaerythritol

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 August - 29 December 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted in accordance with International guidelines and in accordance with GLP. All guideline validity criteria were met.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark and under nitrogen.
- Stability under test conditions: Asumed stable .
- Solubility and stability of the test substance in the solvent/vehicle: Acetone stock solution prepared at 300 mg/mL
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: N/A

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: N/A
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: N/A
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Solubilised in an acetone stock solution which was incorporated into quartz sand and mixed with the artificial soil.

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): N/A

OTHER SPECIFICS: No

Analytical monitoring:

no

Details on sampling:

N/A

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of application to filter paper (if used): N/A
- Method of mixing into soil (if used): An aliquot (16 mL) of an acetone stock solution was added to quartz sand and mixed. The sand was incorportated into the artificial soil and homgenised.
- Method of application to soil surface (if used): N/A
- Controls: Solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone
- Concentration of vehicle in test medium (stock solution and final test solution): 16 mL acetone added to 4720 g soil (dry weight) i.e. 0.003 % v/w.
- Evaporation of vehicle before use: Not reported

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia foetida (redworm)
- Source: Chenggong Earthworm Raising Farm, Dongli District, Tianjing, China.
- Age at test initiation (mean and range, SD): > 2 months old
- Weight at test initiation (mean and range, SD): 300 - 430 g

ACCLIMATION
- Acclimation period: Not reported
- Acclimation conditions (same as test or not): Not reported
- Health during acclimation (any mortality observed): Not reported

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

28 d

Test temperature:

18.16 - 19.50 ºC

pH:

5.53 - 5.62

Moisture:

35 %

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Container measuring 19 cm x 12 cm x 9 cm (H x L x W)
- Amount of soil or substrate: 750 g wet weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 8
- No. of replicates per control: 0
- No. of replicates per vehicle control: 8

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Artificial soil prepared as per OECD guidance. Adjusted and maintained at 35 % moisture content with deionised water for the duration of the test.

OTHER TEST CONDITIONS
- Photoperiod: 16:8 light: dark
- Light intensity: 400-800 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Adult mortality, weight and sub-lethal effects, reproduction

VEHICLE CONTROL PERFORMED: Yes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: N/A, limit test.
- Justification for using less concentrations than requested by guideline: Limit test
- Range finding study: No
- Test concentrations: N/A
- Results used to determine the conditions for the definitive study: N/A

Nominal and measured concentrations:

Nominal: 1000 mg/kg dry weight

Reference substance (positive control):

no

Key result

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

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:

reproduction

Details on results:

- Mortality at end of exposure period: No adult mortality was observed during the test.
- Total mass of adults at beginning of test: 300-430 g
- Changes in body weight of live adults (% of initial weight) at end of exposure period: +12 % and +7 % in the control and 1000 mg/kg groups, respectively
- No. of offspring produced: 51.8 and 42.1 for the control and 1000 mg/kg groups, respectively (mean values).
- No. of unhatched cocoons: Not reported
- Morphological abnormalities: Some cocoons in the 1000 mg/kg test grup looked smaller vs the control group.
- Behavioural abnormalities: No
- Other biological observations: No

Table 1       Earthworm weights

 

Concentration (mg/kg dry soil)	Mean weight of earthworms ± sd (g)		% Increase
	Day 0	Day 28
Control	338.0 ± 36.9	378.2 ± 62.6	+12
1000	343.9 ± 34.0	368.4 ± 46.3	+7

 

Table 2       Adult earthworm mortality

 

Concentration (mg/kg dry soil)	Mean surviving adults	Mean % mortality	Sub-lethal effects
Control	10	0	None
1000	10	0	None

 

Table 3       Juvenile worms

 

Concentration (mg/kg dry soil)	Rep	Adult worm number	Cocoon number	Total juvenile worms	Mean number of juvenile worms ± sd	Reproductive abnormalities
Control	1	10	16	47	51.8 ± 14.6	None
	2	10	17	50
	3	10	15	42
	4	10	13	37
	5	10	13	42
	6	10	23	73
	7	10	24	76
	8	10	14	47
1000	1	10	15	44	42.1 ± 7.2	Some cocoons looked smaller vs control
	2	10	16	46
	3	10	19	55
	4	10	11	30
	5	10	13	42
	6	10	14	43
	7	10	14	39
	8	10	13	38

Validity criteria fulfilled:

yes

Conclusions:

The 28 day LC50 was > 1000 mg/kg dry soil weight. The 28 day NOEC for reproduction was 1000 mg/kg dry soil weight.

Executive summary:

OECD 222 (2012) - In a 28-day reproduction toxicity study, earthworms (Eisenia foetida) were exposed to Tetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid at a nominal concentration of 1000 mg a.i./kg dry weight of artificial soil in a limit test. The experiment was carried out in accordance with the OECD 222 testing guideline. 

 

The test item was solubilized in acetone, an aliquot of which was mixed with quartz sand and incorporated in the artificial soil, prepared in accordance with the testing guideline. The soil was adjusted and maintained at 35 % moisture content throughout the testing period using deionized water. A solvent control was also prepared and run in parallel with the treated soil group.

 

At Day 28, the number of surviving adults was determined. It was noted that there was no mortality observed in any of the control or test replicates. The LC₅₀ was therefore estimated to be >1000 mg/kg dry weight soil. No sub-lethal effects or significant weight gain changes were observed in the treated group versus the control.

 

The number of cocoons and juveniles produced in the control and test group was counted. No adverse effects on reproduction was observed and the NOEC was concluded to be 1000 mg/kg dry soil.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic reproduction toxicity study with earthworms.

Description of key information

28 d LC₅₀: >1000 mg/kg dw soil; OECD 222; Hairong, Y. (2012)

28 d NOEC (reproduction): => 1000 mg/kg dw soil; OECD 222; Hairong, Y. (2012)

Additional information

OECD 222 (2012) - In a 28-day reproduction toxicity study, earthworms (Eisenia foetida) were exposed to Tetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid at a nominal concentration of 1000 mg a.i./kg dry weight of artificial soil in a limit test. The experiment was carried out in accordance with the OECD 222 testing guideline. 

 

The test item was solubilized in acetone, an aliquot of which was mixed with quartz sand and incorporated in the artificial soil, prepared in accordance with the testing guideline. The soil was adjusted and maintained at 35 % moisture content throughout the testing period using deionized water. A solvent control was also prepared and run in parallel with the treated soil group.

 

At Day 28, the number of surviving adults was determined. It was noted that there was no mortality observed in any of the control or test replicates. The LC₅₀ was therefore estimated to be >1000 mg/kg dry weight soil. No sub-lethal effects or significant weight gain changes were observed in the treated group versus the control.

 

The number of cocoons and juveniles produced in the control and test group was counted. No adverse effects on reproduction was observed and the NOEC was concluded to be 1000 mg/kg dry soil.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic reproduction toxicity study with earthworms.