Amines, N-C10-16-alkyltrimethylenedi-, reaction products with chloroacetic acid

Toxicity to soil microorganisms

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil microorganisms

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-06-26 to 2007-07-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 216 (Soil Microorganisms: Nitrogen Transformation Test)

Version / remarks:

2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 217 (Soil Microorganisms: Carbon Transformation Test)

Version / remarks:

2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.21 (Soil Microorganisms: Nitrogen Transformation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.22 (Soil Microorganisms: Carbon Transformation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not required

Vehicle:

no

Details on preparation and application of test substrate:

AMENDMENT OF SOIL
- Type of organic substrate:
Nitrogen transformation experiment: Sieved and spiked soil was amended by the addition of powdered plant material (lucerne-grass-green meal) at a plant-soil ratio of 5 g plant per kilogram of soil (dry mass).

APPLICATION OF TEST SUBSTANCE TO SOIL
- Method: The quantity of test item required to obtain the desired concentrations was filled up to 60 mL with water (the amount of water needed to adjust the test substrate to 52 - 58 % water holding capacity), added to the soil, mixed thoroughly and placed into the test container.

Test organisms (inoculum):

soil

Total exposure duration:

28 d

Test temperature:

19.5-20.0°C

Moisture:

55 % of the WHCmax with a range of 3 %; controlled weekly by weighing; adjustment using deionised water as needed

Organic carbon content (% dry weight):

0.93

Nitrogen content (% dry weight):

0.09

Details on test conditions:

TEST SYSTEM
- Amount of soil: 500 g (Nitrogen transformation), 1000g (Carbon transformation)
- No. of replicates per concentration: 3
- No. of replicates per control: 4

SOURCE AND PROPERTIES OF SUBSTRATE
- Geographical reference of sampling site: D-57377 Schmallenberg, Germany
- History of site: Agricultural field with no culture
- Treatments with pesticides or fertilizers: No plant protection product for at least one year prior to sampling and no organic or mineral fertilisation six or three months prior to sampling
- % sand/silt/clay: 71 / 24 / 5
- pH (in water): 5.7

EFFECT PARAMETERS MEASURED:
Nitrogen transformation:
Nitrate: photometrically (Spectroquant® NOVA 400) on day 0 and 28
Nitrate was extracted from soil by shaking samples (10 g dry mass) with 0.1 M KCl solution at a ratio of 5 mL of KCl solution per gram dry weight for 60 minutes at 150 rpm. The mixtures were filtered and the liquid phases were photometrically analysed for nitrate.
Carbon transformation:
Glucose-induced respiration rates: oxygen consumption (Sapromat® Voith Inc.) on day 0 and 28
The glucose amended soil samples were incubated in an apparatus for continuous measurement of respiration rates (Sapromat® Voith Inc.) at 20 ± 2 °C. The oxygen consumed was measured for 12 consecutive hours.

For further details, refer to section "Any other information on materials and methods incl. tables".

Nominal and measured concentrations:

Nominal concentrations: 8.2, 20.5, 51.2, 128, 320, 800, 2000, and 5000 mg test item/kg, corresponding to 1.64, 4.10, 10.24, 25.6, 64, 160, 400, and 1000 mg a.s./kg dry mass

Reference substance (positive control):

no

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: nitrate formation rate, respiration rate

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

64 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

nitrate formation rate

Remarks on result:

other:

Remarks:

Recalculated value

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: nitrate formation rate, respiration rate

Reported statistics and error estimates:

NOEC: William’s test (one-sided)
Concentration-effect relationship: Probit-analysis assuming log-normal distribution (Computer software ToxRat Professional version 2.09 (release 30.10.2005), ToxRat® Solutions GmbH)

Table A7.5.1.1-5:Mean nitrate content [mg/kg] and deviation from control [%]; a.s. = active substance; concentrations given as nominal concentration per kg dry soil; significant differences were not detected compared to control (Williams t-test, p = 0.05, one-sided).

	Control	1.64 mg a.s./kg	4.10 mg a.s./kg	10.2 mg a.s./kg	25.6 mg a.s./kg	64 mg a.s./kg	160 mg a.s./kg	400 mg a.s./kg	1000 mg a.s./kg
Nitrate [mg/kg]
Test start	78.8	95.0	95.0	121.7	123.3	105.0	106.7	110.0	93.3
Day 28	202.5	190.0	210.0	203.5	226.7	240.0	243.3	296.7	428.3
Deviation [%]		–6.2	3.7	0.4	11.9	18.5	20.2	46.5	111.5

 

Table A7.5.1.1-6:Mean short-term respiration rate ([mg O₂/(kg × h)] and deviation from control [%]; a.s. = active substance; SIR = Substrate induced respiration; concentrations given as nominal concentration per kg dry soil; significant differences were not detected compared to control (Williams’ t-test, p = 0.05, one-sided).

	Control	1.64 mg a.s./kg	4.10 mg a.s./kg	10.2 mg a.s./kg	25.6 mg a.s./kg	64 mg a.s./kg	160 mg a.s./kg	400 mg a.s./kg	1000 mg a.s./kg
SIR [mg O2/(kg × h)]
Test start	5.17	4.51	5.42	6.06	7.15	6.07	5.91	7.34	5.82
Day 28	3.88	4.13	3.11	3.02	3.35	3.52	3.97	6.87	10.22
Deviation [%]		6.4	–19.8	–22.2	–13.7	–9.3	2.3	77.1	163.4

 

Validity criteria fulfilled:

yes

Conclusions:

The NOEC of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid for both, nitrogen and carbon transformation by soil microorganism was >/= 1000 mg a.i./kg dry soil. The EC50 was > 1000 mg a.i./kg dry soil.

Executive summary:

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid (20% a.i., as manufactured) on nitrogen and carbon transformation by soil microorganisms was investigated according to the OECD guidelines 216 and 217. The test item was incorporated into loamy sand soil at nominal concentrations of 1.64, 4.10, 10.24, 25.6, 64, 160, 400, and 1000 mg a.s./kg dry mass at test start. The effects on nitrogen transformation and short-term respiration were determined 28 days after start of incubation.

With respect to the nitrogen transformation capacity and short-term respiration, no inhibitory effect of the test item was observed. The NOEC for both, nitrogen and carbon transformation was >/= 1000 mg a.i./kg dry soil. The EC50 was > 1000 mg a.i./kg dry soil.

Description of key information

NOEC (28 d): 64 mg/kg soil dw (nominal, nitrogen and carbon transformation by soil microorganisms, OECD guidelines 216 and 217)

Key value for chemical safety assessment

Short-term EC50 for soil microorganisms:

1 000 mg/kg soil dw

Long-term EC10 or NOEC for soil microorganisms:

64 mg/kg soil dw

Additional information

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid (20% a.i., as manufactured) on nitrogen and carbon transformation by soil microorganisms was investigated according to the OECD guidelines 216 and 217. The test item was incorporated into loamy sand soil at nominal concentrations of 1.64, 4.10, 10.24, 25.6, 64, 160, 400, and 1000 mg a.i./kg dry mass at test start. The effects on nitrogen transformation and short-term respiration were determined 28 days after start of incubation.

With respect to the nitrogen transformation capacity and short-term respiration, no inhibitory effect of the test item was observed. The NOEC for both, nitrogen and carbon transformation was >/= 1000 mg a.i./kg dry soil. The EC50 was > 1000 mg a.i./kg dry soil. A recalculation revealed that inhibitory effect can be seen at 160 mg a.i./kg dry soil. Therefore, the NOEC is 64 mg a.i./kg dry soil.