Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates

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:

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

Adequacy of study:

key study

Study period:

2007-12-12 to 2008-02-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP, All validity criteria fulfilled, complete identification of test substance

Justification for type of information:

The category and read-across approach are justified in both documents, please find this further information in the attached documentation part in the dedicated ESR for terrestrial toxicity "Supporting Read-Across document".

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

DIN ISO 11268-2:1998 (E), BBA Guideline

Deviations:

yes

Remarks:

Fresh Lufa 2.2 was used instead of artificial soil

Principles of method if other than guideline:

LUFA 2.2 soil, classified as a loamy sand, was used rather than the soil recommended by the OECD guideline as it is considered a reasonable worst case and this laboratory has experience with this soil. Moreover, the OECD soil contains 20 % kaolinite, a clay type which may influence the test results if used in conjunction with a cationic surfactant. Fresh rather than dried soil was used as this is closer to the soil that would be found in the field and is therefore expected to be more environmentally realistic.

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Vapour pressure: < 1 hPa (20 °C)
- Water solubility (under test conditions): Insoluble

Analytical monitoring:

no

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): A mixture comprised of 10 g quartz sand with the necessary quantity of the test item was prepared per treatment. The mixture was thoroughly mixed with the pre-moistened soil with a long handled blender for at least four minutes. Afterwards the soil was moistened to 54 % of its maximum water holding capacity.
- Control: Fresh LUFA 2.2 standard soil moistened with demineralised water without test or reference item
- Vehicle control: Fresh LUFA 2.2 standard soil mixed with quartz sand and moistened with demineralised water without test or reference item

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: Eisenia fetida SAVIGNY 1826, subspecies: Eisenia fetida fetida
- Source: Own breeding (Source: Bundesanstalt fuer Verbraucherschutz und Landwirtschaft (BVL), Messeweg 11/12, D-38104 Braunschweig)
- Age at test initiation (mean and range, SD): 9-10-month, not variating more than 1 month
- Weight at test initiation (mean and range, SD): 0.53 +/- 0.04 g


ACCLIMATION
- Acclimation period: 2 days prior to test start
- Acclimation conditions (same as test or not):same as test
- Health during acclimation (any mortality observed): healthy

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

55 d

Remarks:

removement of adults after 28 days

Test temperature:

20 +/- 2 °C

pH:

Table 1: pH-Values of the Test Media
Measurement of pH and of test medium was based on mixed samples of all replicates per treatment
and control.
Application rate pH-value
[mg/kg soil DW] day 0 day 56
Control 5.618 6.52
Vehicle Control 5.710 6.72
125 5.773 6.66
250 5.749 6.67
500 5.779 6.72
1000 5.914 6.75

Moisture:

Table 1: Moisture of the Test Media
Measurement of moisture content of test medium was based on mixed samples of all replicates per
treatment and control.
Application rate Moisture [%] of dry weight

[mg/kg soil DW] day 0 day 56
Control 22.77 27.21
Vehicle Control 22.83 28.25
125 26.02 27.98
250 22.51 28.53
500 23.82 30.92
1000 24.46 30.57

Details on test conditions:

TEST SYSTEM
- Test container (material, size): round plastic boxes, ID: 15.0 cm, bottom surface area: 177 cm3, height: 14 cm; transparent and perforated lids enabled sufficient gas exchange and light penetration and prevented the test substrate from drying
- Amount of soil or substrate: 600 g DW
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- No. of replicates per vehicle control: 4


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Germany / Rheinland Pfalz / Hanhofen, "Grosser Striet", No. 585, owner: Appel, Hanhofen
- Pesticide use history at the collection site: None (former 4 years)
- Collection procedures: Sampling date 2007-10-30, batch F2.24407
- Sampling depth (cm): approximately 20 cm
- Soil texture (if natural soil)
- % sand: 0.63 - 2.0 mm: 0.7 %, 0.2 - 0.63 mm: 44.4 %, 0.063 - 0.2 mm: 35.7 %
- % silt: 0.02 - 0.063 mm: 5.2 %, 0.006 - 0.02 mm: 3.9 %, 0.002 - 0.006 mm: 3.4 %
- % clay: 6.7
- Soil taxonomic classification: loamy sand
- Organic carbon (%): 2.29
- Maximum water holding capacity: 49.7 g/100 g DW
- CEC: 10 mval/100 g
- Pretreatment of soil: Drying (room temperature, until sievable), presieving 10 mm, last sieving 2 mm, moistening at approximately 19 % of water holding capacity
- Storage (condition, duration): 4 - 7 °C for 43 days prior to use



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


EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Adult mortality, biomass, behaviour and pathological symptoms after 28 days
Reproductive output after 55 days

VEHICLE CONTROL PERFORMED: yes


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: Due to the results of the preliminary test no effects were expected.
- Range finding study (NON-GLP)
- Test concentrations: 100 - 500 - 1000 - 2000 mg/kg soil DW
- Results used to determine the conditions for the definitive study: No mortality or significant change of live weight was determined.

Nominal and measured concentrations:

Nominal application rates: 125 - 250 - 500 - 1000 mg/kg soil dry weight (DW)

Reference substance (positive control):

yes

Remarks:

1.0 - 2.5 - 5.0 mg Carbendazim/kg soil DW

Duration:

55 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Reproduction, mortality, body weight

Details on results:

- Mortality at end of exposure period: No significant mortality (> 10 %) was observed in any of the tested test item and control groups.
- Total mass of adults at beginning of test: 0.53 g (mean)
- Changes in body weigth of live adults (% of initial weight) at end of exposure period:
Control: 6.3 +/- 3.44, Vehicle Control: 12.8 +/- 7.50, 125 mg/kg soil DW: 7.9 +/- 5.75, 250 mg/kg soil DW: 6.3 +/- 7.72, 500 mg/kg soil DW: 12.1 +/- 3.60, 1000 mg/kg soil DW: 9.1 +/- 2.86
- No. of offspring produced:
Control: 291 +/- 46.1, Vehicle Control: 344 +/- 11.5, 125 mg/kg soil DW: 333 +/- 82.9, 250 mg/kg soil DW: 271 +/- 37.7, 500 mg/kg soil DW: 310 +/- 25.7, 1000 mg/kg soil DW: 297 +/- 32.3
- No. of unhatched cocoons: Not determined
- 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: EC50 (Reproduction) = 1.19 mg/kg

Reported statistics and error estimates:

One Way Analysis of Variance (ANOVA and Dunnett's Method) was carried out for determination of statistically significant differences between control and vehicle control and further between pooled control and test item concentrations. Prior to Analysis of Variance, a Normality Test and an Equal Variance Test were conducted. P-values for both normality and equal variance test are 0.05. The a-value for ANOVA test (acceptable probability of incorrectly concluding that there is a difference) is a = 0.05.

Adult mortality:

No significant mortality (> 10 %) was observed in any of the tested doses of the test item and the control groups.

Behaviour and Pathological Symptoms:

No pathological and behavioural symptoms were found at the tested application rates and control groups.

Live weight:

At test start earthworms had a mean weight of 0.53 - 0.6 g. The body weights of earthworms in tested doses and control groups increased during test period. Effects on body live weights after 28 days were tested for statistical significance by One Way Analysis of Variance (ANOVA). There was no statistically significant difference when compared to the control.

Reproduction rate:

The average number of juveniles of treated groups ranged between 271 and 333 individuals. Mean number of juveniles found in control groups were between 291 and 344. The coefficient of variation calculated for the reproduction of control was lower than 30 %. No statistically significant difference to the control groups was determined (ANOVA).

Validity criteria fulfilled:

yes

Conclusions:

The test item did not induce significant effects on adult worms in terms of reproduction, mortality, body weight in natural soil (fresh LUFA 2.2) on Eisenia fetida fetida. The NOEC was 1000 mg/kg soil DW.

Executive summary:

Effects of N-(hydrogenated tallow alkyl)trimethylenediamines (Batch no. S000905) on mortality, biomass and the reproductive potential of the earthworm species Eisenia fetida fetida (Savigny) were determined according to the guidelines DIN ISO 11268-2 (1998), OECD 222 (2004) and BBA (1994) from December 12, 2007 to February 5, 2008 at Dr.U.Noack-Laboratorien in D-31157 Sarstedt. The study was conducted under static conditions over 8 weeks with the four different application rates 125 - 250 - 500 - 1000 mg/kg soil dry weight (DW) mixed with natural soil (fresh LUFA 2.2). Eighty adult earthworms with clitellum in control and 40 test organisms in vehicle control and test item concentrations with an individual weight of 0.35 to 0.60 g, divided into eight and four replicates, respectively were inserted. For statistical evaluation control and vehicle control were pooled because analysis of variance showed no significant differences between controls.

After 28 days no significant differences occurred concerning mortality, pathological symptoms or behaviour of the adult earthworms at all tested application rates and in the control groups. The individual body live weights increased in the control as well as in the test item groups. After further 28 days the reproduction (average number of juveniles) ranged from 291 to 344 in the control groups and from 271 to 333 juveniles in the test item groups, respectively (Table 7). There was no statistically significant change of mortality, biomass and reproduction compared to the pooled control at all application rates (Table 1). The NOEC and LOEC are summarized in Table 2. The validity criteria recommended by the test guidelines were fulfilled.

Table 1:     Summary of Observed Effects in the Tested Doses

Effects	N-(hydrogenated tallow alkyl)trimethylenediamines
[mg test item/kg soil DW]	125	250	500	1000
Mortality of adult earthworms	No	No	No	No
Reduction of body weight versus  control	No	No	No	No
Reduced reproduction rate    (Number of juveniles)	No	No	No	No
Summary of all test effects	No	No	No	No

No     =    no statistical significance after comparison with pooled controls (ANOVA, Dunnett's method)

Table 2:     NOEC and LOEC for Reproduction, Mortality and Body Weight

N-(hydrogenated tallow alkyl)trimethylenediamines
	Application rate [mg/kg soil DW]
NOECreproduction, mortality, body weight	1000
LOECreproduction, mortality, body weight	> 1000

NOEC for N-(hydrogenated tallow alkyl)trimethylenediamines with respect to mortality, body weight and reproduction of Eisenia fetida fetida,

tested in natural soil (fresh LUFA 2.2), was 1000 mg/kg soil DW.

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

Study period:

October 2020

Justification for type of information:

REPORTING FORMAT FOR THE READ-ACROSS APPROACH
The category and read-across approach are justified in both documents, please find this further information in the attached documentation part in this dedicated ESR for terrestrial toxicity.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other:

Version / remarks:

REACH TGD, Chapter R.6: QSARs and grouping of chemicals.

Deviations:

not specified

GLP compliance:

no

Key result

Duration:

55 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

reproduction, mortality, body weight

Description of key information

No toxicity of Amines, N-C16-18-alkyl (evennumbered) propane-1,3-diamine (CAS  133779-11-0) to earthworms was observed at the highest concentration tested i.e. 1000 mg/kg sediment dw.

Key value for chemical safety assessment

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

1 000 mg/kg soil dw

Additional information

One chronic earthworm test (OECD 222, GLP, reliability 2) has been performed with Amines, N-C16-18-alkyl (evennumbered) propane-1,3-diamine (CAS 133779-11-0). Effects on mortality, biomass and reproduction were not observed. Therefore the NOEC earthworm is equal to the highest test concentration of 1000 mg a.i./kg soil dw after 56 days of exposure.

It is considered to be justified to use these results also for other diamines and the registered substance (CAS 1313206 -64 -2) because only a limited difference is expected for the terrestrial compartment because the main exposure route for sorbing substances will be via ingestion and because the main driver in diamines sorption is ionic interaction.

As confirmed by the physico-chemical properties of diamines acetates, the read-acoss from diamines is justified. Indeed, the presence of acetates has no effect on the fate of diamines acetates in the environment and thus the sorption to soil is expected to be similar between diamines and diamines acetates. Thus these toxicity data obtained for the soil compartment can be used as read-across for Amines, N-(C16 -18 and C18 -unsatd. alkyl)trimethylenedi-, diacetates (CAS 1313206 -64 -2).

 

(see the read-across justification in the attached justification for read-across).