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

Toxicity to terrestrial plants

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to terrestrial plants: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-06-19 to 2007-07-06

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)

Version / remarks:

adopted 19 July 2006

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not required

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: Incorporation into soil as an aqueous solution

Species:

Lactuca sativa

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Lettuce
- Plant family: Asteraceae
- Source of seed: Raiffeisen Naturkraft Kiepenkerl

Species:

Phaseolus aureus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Mung bean
- Plant family: Fabaceae
- Source of seed: Carl Sperling & Co. Lüneburg, Charge 576, D 6210 H

Species:

Raphanus sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Radish
- Plant family: Brassicaceae
- Source of seed: Raiffeisen Naturkraft Kiepenkerl

Species:

Sinapis alba

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: White mustard
- Plant family: Brassicaceae
- Source of seed: Landesinstitut für Landwirtschaftliche Qualitätskontrolle

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oat
- Plant family: Poaceae
- Source of seed: Raiffeisen Genossenschaft Nordwest eG Certification authority Münster - D 993043 D/MS 1160/006

Species:

Triticum aestivum

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Common wheat
- Plant family: Poaceae
- Source of seed: Saatgutveredelung Nueckel, 57392 Schmallenberg-Winkhausen

Test type:

seedling emergence toxicity test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

14 d

Remarks:

14 days after emergence of 50 % of the control seedlings

Test temperature:

19–21 °C

pH:

5.7

Moisture:

60–90 % humidity

Details on test conditions:

TEST SYSTEM
- Test container (type, material, size): Round containers of nonporous plastic with a diameter of 85–95 mm, filled with ca. 280 g of moist soil (natural sandy soil)
- Method of seeding: Five seeds were planted in each replicate within 24 h after incorporation of the test item
- No. of seeds per container: 5
- No. of replicates per treatment group: 4
- No. of replicates per control: 4


SOURCE AND PROPERTIES OF SUBSTRATE
The soil used in the test was a natural sandy soil (Certified RefeSol 01-A; batch IME-01, loamy sand [DIN], Org C: 0.93 %, pH 5.7, clay: 5 %). The soil was sieved to 2 mm. The soil was not sterilized. The soil has been stored outdoor in high grade stainless steel basins with drainage and ground contact at the test facility.

NUTRIENT MEDIUM (if used)
- Description: Continuous bottom watering of the test container via glass fibre wicks. The water was amended with fertilizer (COMBO Grünpflanzendünger)

GROWTH CONDITIONS
- Photoperiod: 16 h light/day
- Light intensity and quality: light intensity of > 7000 lx (light colour 25, universal white)
- Day/night temperatures: 19-21°C
- Relative humidity (%): 60–90 % humidity
- Wind velocity: Not applicable
- Watering regime and schedules: Continuous
- Any pest control method/fertilization (if used): no

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Fresh shoot biomass
Pathological symptoms
Seedling emergence
At growth day 14, all seedlings were counted and the aboveground biomass was measured. For this, the wet mass of the plants was measured immediately after harvesting

Nominal and measured concentrations:

Nominal concentrations: 0, 8.2, 20.5, 51.2, 128, 320, 800, 2000, and 5000 mg/kg corresponding to 1.64, 4.10, 10.2, 25.6, 64, 160, 400, and 1000 mg a.s./kg dry mass

Reference substance (positive control):

yes

Remarks:

trichloroacetate; A test with the reference substance is regularly performed at the test facility once a year, thus fulfilling guideline recommendations.

Key result

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

363 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks on result:

other: 95% CL 316–394 mg/kg soil dw

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

1 064 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Remarks on result:

other: 95% CL 230–4902 mg/ kg soil dw

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Avena sativa

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Sinapis alba

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Sinapis alba

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

>= 400 - <= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Triticum aestivum

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Triticum aestivum

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

other: Avena sativa, Phaseolus aureus, Raphanus sativus, Triticum aestivum

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

other: Lactuca sativa, Sinapis alba

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

64 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

other: Avena sativa, Sinapis alba

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

400 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Triticum aestivum

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

160 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

< 1.64 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

10.2 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Details on results:

for details see section "Any other information on results incl. tables"

Results with reference substance (positive control):

- Results with reference substance valid?
Results are in agreement with experience EC50-values in the testing facility.

Reference substance: Seedling emergence:
Avena sativa: Control: 97.5 %; 100 mg TCA/kg: 85.0 %
Lactuca sativa: Control: 85.0 %; 100 mg TCA/kg: 62.5 %
Phaseoulus aureus: Control: 92.5 %; 100 mg TCA/kg: 77.5 %
Raphanus sativus: Control: 97.5 %; 100 mg TCA/kg: 95.0 %
Sinapis alba: Control: 90.0 %; 100 mg TCA/kg: 80.0 %
Triticum aestivum: Control: 95.0 %: 100 mg TCA/kg: 90.0 %
EC50 values could not be computed.

Reference substance: EC50 for growth (fresh mass per plant):
Avena sativa: 6 mg/kg
Lactuca sativa: 55 mg/kg
Phaseoulus aureus: 21 mg/kg
Raphanus sativus: 175 mg/kg
Sinapis alba: 46 mg/kg
Triticum aestivum: 1 mg/kg

Reported statistics and error estimates:

The percent inhibition of seedling emergence for each plant species was calculated as an absolute value and in comparison to the control. Survival of emerged seedlings was calculated as an absolute value. The percent inhibition of fresh weight was calculated in comparison to the control. All statistical analyses (Mann-Whitney U-Test, Fisher’s Exact Binomial Test, or Welch’s t-test for comparison of controls; the Chi² 2 × 2 Table Test with Bonferroni Correction, Williams’ test, or Welch’s t-test for the NOEC calculation; EC50 estimation, as appropriate) were performed with the computer software ToxRat Professional by ToxRat® Solutions GmbH. For the EC50 calculation probit analysis assuming log-normal distribution of the data was applied.

Table A7.5.1.3 -4: Pathological symptoms [% plants]; a = discolouration, b = deformation, c = necrosis, d = dead plant.

Test species	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
Avena sativa	–	–	–	–	–	–	–	–	–
Lactuca sativa	5 % d	5 % d	-	10 % d	15 % d	–	–	–	5 % d
Phaseolus aureus	–	–	–	–	–	–	10 % d	–	-
Raphanus sativus	–	–	–	–	–	–	–	–	–
Sinapis alba	–	5 % d	10 % d	–	–	–	5 % d	–	–
Triticum aestivum	–	–	–	–	–	–	-	–	–

 

Table A7.5.1.3-5:Emergence rate at test end [%].TI = Test item.

Test species	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
Avena sativa	95	100	100	100	85	95	95	90	95
Lactuca sativa	85	80	80	80	90	75	55	55	45
Phaseolus aureus	100	100	100	100	95	100	100	100	100
Raphanus sativus	95	95	100	100	95	100	100	100	100
Sinapis alba	100	85	95	90	95	90	80	75	85
Triticum aestivum	95	75	100	100	95	95	85	100	90

 

Table A7.5.1.3-6:Emergence related to solvent control at test end [%]; *: significant when compared with control.

Test species	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
Avena sativa	–5	–5	–5	11	0	0	5	0
Lactuca sativa	6	6	6	–6	12	35*	35*	47*
Phaseolus aureus	0	0	0	5	0	0	0	0
Raphanus sativus	0	–5	–5	0	–5	–5	–5	–5
Sinapis alba	15	5	10	5	10	20*	25*	15*
Triticum aestivum	21*	–5	–5	0	0	11	–5	5

 

Table A7.5.1.3-7:Fresh weight of the shoots.Mean values ± SD[g]TI = Test item.

Test species	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
Avena sativa	0.840	0.933	0.862	0.880	1.073	0.973	0.833	0.815	0.671
Lactuca sativa	1.008	1.277	1.269	1.403	1.227	1.060	1.051	0.405	0.080
Phaseolus aureus	1.699	1.112	0.986	0.959	1.011	0.780	0.936	0.925	0.895
Raphanus sativus	1.727	1.668	1.593	1.628	1.304	1.439	1.601	1.500	0.904
Sinapis alba	0.850	1.046	1.012	1.040	1.247	1.135	1.050	1.223	0.419
Triticum aestivum	0.718	0.776	0.789	0.761	0.761	0.772	0.779	0.719	0.551

 

Table A7.5.1.3-8:Growth inhibition related to solvent control at test end [% FM];TI = Test item; negative value = growth stimulation; *: significant when compared with the control.

Test species	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
Avena sativa	–11	–3	–5	–28	–16	1	3	20*
Lactuca sativa	–27	–26	–39	–22	–5	–4	60*	92*
Phaseolus aureus	35	42	44	40	54	45*	46*	47*
Raphanus sativus	3	8	6	25*	17*	7*	13*	48*
Sinapis alba	–23	–19	–22	–47	–34	–24	–44	51*
Triticum aestivum	–8	–10	–6	–6	–8	–9	0	23

 

Table A7.5.1.3-9:NOECs and EC₅₀values for emergence [mg/kg TM];NOEC = no observed effect concentration; EC = effect concentration; CL = confidence limits; n.d. = not determined for mathematical reasons; nominal concentrations were multiplied with a factor of 0.714 to take application loss into account.

Test species	Avena sativa	Lactuca sativa	Phaseolus aureus	Raphanus sativus	Sinapis alba	Triticum aestivum
NOEC	≥ 1000	64	≥ 1000	≥ 1000	64	≥ 1000
EC50(95% CL)	≥ 1000 (n.d.)	1064 (230–4902)	≥ 1000 (n.d.)	≥ 1000 (n.d.)	≥ 1000 (n.d.)	≥ 1000 (n.d.)

 

Table A7.5.1.3-10:NOEC and EC₅₀values for growth inhibition on the basis of fresh mass [mg/kg TM]; NOEC = no observed effect concentration; EC = effect concentration; CL = confidence limits; n.d. = not determined for mathematical reasons; nominal concentrations were multiplied with a factor of 0.714 to take application loss into account.

Test species	Avena sativa	Lactuca sativa	Phaseolus aureus	Raphanus sativus	Sinapis alba	Triticum aestivum
NOEC	400	160	<1.64	10.2	400	≥ 1000
EC50(95% CL)	≥ 1000 (n.d.)	363 (316–394)	≥ 1000 (n.d.)	≥ 1000 (n.d.)	400–1000 (n.d.)	≥ 1000 (n.d.)

 

Validity criteria fulfilled:

yes

Conclusions:

Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid had a mild inhibitory effect on growth in five of the six tested plant species. With respect to seedling emergence, there was only a mild inhibitory effect in two of the six tested plant species. The EC50 values were between 363 mg a.i./kg to 1000 mg a.i./kg, depending on the species.

Executive summary:

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid (20% a..i., as manufactured) on the emergence and growth of terrestrial plant seedlings was investigated according to the OECD guideline 208. The following test species were used: Avena sativa (oat), Lactuca sativa (lettuce), Phaseolus aureus (mung bean), Raphanus sativus (radish), Sinapis alba (mustard), and Triticum aestivum (wheat).

The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 0, 1.64, 4.10, 10.2, 25.6, 64, 160, 400, and 1000 mg a.s./kg dry mass, respectively. The concentration of 1000 mg/kg corresponds to the limit test concentration recommended by OECD-guideline 208. The test item was incorporated into the test soil at the beginning of the experiment, and the effects on seedling emergence and growth were determined 14 days after emergence of 50 % of the control seedlings. Four replicates were prepared for the control and per concentration.

There was no concentration dependent effect on seedling emergence of Avena sativa, Phaseolus aureus, Raphanus sativus, and Triticum aestivum up to 1000 mg a.s./kg, the highest concentration tested. Thus, the NOEC for these tested species was found to be ≥ 1000 mg a.s./kg dry soil. The EC50 was found to be > 1000 mg a.s./kg dry soil.

There was a concentration dependent effect on seedling emergence of Lactuca sativa and Sinapis alba starting at 400 mg a.s./kg (NOEC = 160 mg a.s./kg).

However, the EC50 for Lactuca sativa was found to be 1062 mg a.s./kg (by extrapolation), and that for Sinapis alba > 1000 mg a.s./kg dry soil.

There were no statistically significant effects on growth (on a fresh weight per plant basis) of Triticum aestivum up to the highest test concentration (NOEC ≥ 1000 mg a.s./kg, EC50 > 1000 mg a.s./kg). Concentration related inhibition of plant growth was observed in Avena sativa (NOEC = 400 mg a.s./kg), Phaseolus aureus (NOEC ≤ 1.64 mg a.s./kg), Raphanus sativus (NOEC = 10.2 mg a.s./kg), Lactuca sativa (NOEC = 160 mg a.s./kg), and Sinapis alba (NOEC = 400 mg a.s./kg). However, for the first three of these five species, the effects were only moderate, with the growth rate never being inhibited by more than 50 %. Accordingly, the EC50 values for these tested species were found to be > 1000 mg a.s./kg dry soil. For Lactuca sativa and Sinapis alba, EC50 values of 363 mg a.s./kg and 400–1000 mg a.s./kg, respectively, were found. for the tested species was found to be >/= 1000 mg a.s./kg dry soil.

Reference 2

Endpoint:

toxicity to terrestrial plants: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2014-10-20 to 2014-11-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)

Version / remarks:

adopted 19 July, 2006

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Düsseldorf, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: triplicate samples of the application suspension with the highest item concentration was taken and analyzed analytically

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: The volume of a stock solution (Solution I: 32.0 g test item in 200 mL water; Solution II: 800 mg test item in 100 mL water) required to obtain the desired concentrations in the soil was filled up to 100 mL and mixed throughly with 3200 g dry mass test substrate (adjusted to 7.4% WHC). Subsequently, the test substrate was adjusted to approximately 35% WHC with water and mixed thoroughly. For further moistening to approx. 60 % water holding capacity after sowing.
- Controls: The control consisted of test substrate only which was treated equivalent to the test item concentrations but with water only. Controls were conducted in five replicates.

Species:

Phaseolus aureus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Mung bean
- Plant family: Fabaceae
- Source of seed: Carl Sperling & Co. Lüneburg, Germany, Charge 104
- Seed storage: The seeds were stored in the dark at the test facility at room temperature (20 ± 5 °C) and protected from moisture until use.

Species:

Raphanus sativus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Radish
- Plant family: Brassicaceae
- Source of seed: Quedlinburger Saatgut GmbH, Quedlinburg, Germany, Charge 2213h
- Seed storage: The seeds were stored in the dark at the test facility at room temperature (20 ± 5 °C) and protected from moisture until use.

Test type:

seedling emergence toxicity test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

14 d

Remarks:

14 days after emergence of 50% of the control seedlings.

Post exposure observation period:

no

Test temperature:

20 - 23 °C

pH:

4.9

Moisture:

60 - 90% humidity

Details on test conditions:

TEST SYSTEM
- Testing facility: Plant growth chambers
- Amount of soil: 310 g of moist soil
- Test container: Round containers of nonporous plastic with a diameter of 85–95 mm
- Method of seeding: Five seeds were planted in each replicate right after incorporation of the test item
- No. of seeds per container: 5
- No. of replicates per treatment group: 5
- No. of replicates per control: 5

SOURCE AND PROPERTIES OF SUBSTRATE
The soil used in the test was a natural sandy soil (Certified RefeSol 01-A; batch IME 04, loamy sand [DIN], Org C: 1.09 %, pH (CaCl2) 4.9, clay: 6.1 % [DIN]). The soil was sieved to 2 mm. The soil was not sterilized. The soil has been stored outdoor in high grade stainless steel basins with drainage and ground contact at the test facility.

NUTRIENT MEDIUM
- Description: Continuous bottom watering of the test container via glass fibre wicks. The water was amended with fertilizer as necessary.
COMPO Grünpflanzendünger was used as fertilizer. Manufactory: COMPO GmbH & Co. KG, Münster, Germany. The concentration applied in the test was 1 mL fertilizer per liter water.
Nutrient content of COMPO Grünpflanzendünger
Ammonium nitrogen 37 g/L Iron (Chelate) 0.50 g/L
Nitrate nitrogen 33 g/L Copper (Chelate) 0.02 g/L
Phosphate 30 g/L Manganese (Chelate) 0.20 g/L
Potassium oxide 60 g/L Molybdenum 0.01 g/L
Magnesium oxide 60 g/L Zinc (Chelate) 0.02 g/L
Sulfur 18 g/L Boron 0.10 g/L

GROWTH CONDITIONS
- Photoperiod: 16 hours per day
- Light source: artificial light
- Light intensity and quality: intensity of 244 - 299 µE/(m2/s) (light colour 25, universal white)
- Day/night temperatures: 20 - 23 °C
- Relative humidity: 60 - 90%
- Watering regime and schedules: Continous

EFFECT PARAMETERS MEASURED:
- At test end: growth (on fresh weight basis), pathological symptoms, seedling emergence, seedling survival

Nominal and measured concentrations:

Nominal concentrations: 0 (control), 12.8, 42.2, 139, 459, 1515, and 5000 mg test item/kg dry mass corresponding to 2.50, 8.23, 27.1, 89.5, 295, and 975 mg a.s./kg dry mass
Measured concentrations in application suspension with the highest item concentration (nominal 160 g/L): C12-PDA: 10.1 - 12.0 g/L (101% relative recovery), N-C12 Gly: 3.0 - 3.5 g/L (101.9% relative recovery) N’-C12 Gly: 4.7 - 5.7 g/L (109.4% relative recovery)

Reference substance (positive control):

yes

Remarks:

trichloroacetate; A test with the reference substance is regularly performed at the test facility once a year

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

801 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 295 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Species:

Raphanus sativus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Phaseolus aureus

Duration:

14 d

Dose descriptor:

EC50

Effect conc.:

> 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Details on results:

- Phatological symptoms: None observed.
- Seedling emergence was not affected significantly in any species tested. Germination rate was not inhibited due to the test item up to a concentration of 5000 mg test itemper kg dry soil (NOEC emergence ≥ 5000 mg/kg dry soil), which was the highest concentration tested.
- Post-emergence survival: no statistically significant effect occurs up to a concentration of 5000 mg test item per kg dry soil (NOEC survival ≥ 5000 mg/kg dry soil).
- Biomass on a per plant basis of Phaseolus aureus was not significantly affected by test item up to and including 5000 mg test item per kg soil. The biomass of Raphanus sativus was significantly reduced by the test item starting at 5000 mg test item per kg soil. The NOEC was 1515 mg test item per kg soil, representing 295 mg a.s./kg. The EC50 biomass was found to be 4109 mg test item per kg dry soil (95 % CL: 3321 – 5434 mg/kg), representing 801 mg a.s./kg (95 % CL: 648 – 1060 mg/kg).

Results with reference substance (positive control):

Growth Tests with trichloroacetate and test species Phaseolus aureus (mung bean) and Raphanus sativus (radish). The results of the last reference study are in agreement with experience EC50-values obtained in this institute:
Seedling emergence:
Phaseoulus aureus: Control 100.0 % 100 mg TCA/kg 80.0 %
Raphanus sativus: Control 81.3 % 1000 mg TCA/kg 50.0 %
EC50 values could not be computed.
EC50 for growth:
Phaseoulus aureus: 26 mg/kg (fresh mass per plant)
Raphanus sativus: > 1000 mg/kg (fresh mass per plant)

Reported statistics and error estimates:

The percent inhibition of seedling emergence for each plant species was calculated as an absolute value and in comparison to the control. Survival of emerged seedlings was calculated as an absolute value. The percent inhibition of fresh weight was calculated in comparison to the control. All statistical tests - depending on normal distribution (after Shapiro-Wilks test) and homogeneity of variances (after Levene test) the Fisher`s Exact Binomial Test, Dunnett`s Multiple t-test, or Williams t-test for the NOEC calculation; ECX calculation - were performed with the computer software ToxRat Professional by ToxRat® Solutions GmbH. For the ECX calculation a probit-analysis assuming log-normal distribution of the values was applied.

Table 1:  Emergence rate at test end [%]. TI = Test item; DM = Dry mass; Concentrations given as nominal values.

Test species	Control	12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus	92	96	100	96	100	96	100
Raphanus sativus	76	64	64	72	84	72	72

Table 2:  Emergence inhibition related to control at test end [%]. TI = Test item; DM = Dry mass; Concentrations given as nominal valus; No significant deviation when compared to control (Fisher`s Exact Binomial Test, p = 0.05, one-sided smaller); negative value = emergence stimulation.

Test species		12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus		-4	-9	-4	-9	-4	-9
Raphanus sativus		16	16	5	-11	5	5

Table 3:  Fresh weight of the shoots.Mean values ± SD[g]. TI = Test item; DM = Dry mass; Concentrations given as nominal values.

Test species	Control	12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus	0.849	0.699	0.962	0.647	0.920	0.934	0.721
	±	±	±	±	±	±	±
	0.129	0.106	0.239	0.101	0.088	0.120	0.065
Raphanus sativus	0.841	1.207	0.824	0.954	0.882	0.630	0.380
	±	±	±	±	±	±	±
	0.352	0.234	0.174	0.379	0.320	0.104	0.113

 

Table 4:  Growth inhibition related to control at test end [% FM]. TI = Test item; FM = Fresh mass; DM = Dry mass; Concentrations given as nominal valus; *: significant when compared to control (Williams t-test, p = 0.05, one-sided smaller); negative value = growth stimulation.

Test species		12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus		18	-13*	24	-8	-10	15
Raphanus sativus		-44	2	-13	-5	25	55*

Table 5:  Post-emergence survival rate at test end [%]. TI = Test item; DM = Dry mass. Concentrations given as nominal values.

Test species	Control	12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus	100	96	100	92	100	100	96
Raphanus sativus	89	94	75	78	62	100	94

 

Table 6:  Post-emergence survival inhibition related to control at test end [%]. TI = Test item; DM = Dry mass; No significant deviation when compared to control (Fisher`s Exact Binomial Test, p = 0.05, one-sided smaller); negative value = emergence stimulation. Concentrations given as nominal values.

Test species		12.8 mg TI per kg DM	42.2 mg TI per kg DM	139 mg TI per kg DM	459 mg TI per kg DM	1515 mg TI per kg DM	5000 mg TI per kg DM
Phaseolus aureus		4	0	8	0	0	4
Raphanus sativus		-5	16	13	31	-12	-6

Validity criteria fulfilled:

yes

Conclusions:

Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid has only moderate effects on terrestrial plants under the chosen test conditions. The relevant NOEC for the most sensitive endpoint (growth of Raphanus sativus) was found to be 1515 mg/kg dry soil, representing 295 mg a.s./kg. The EC50 was calculated to be 4109 mg/kg dry soil, representing 801 mg a.s./kg.

Executive summary:

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid on the emergence and growth of terrestrial plant seedlings was investigated according to the OECD guideline 208. The following test species were used: Phaseolus aureus (mung bean) and Raphanus sativus (radish), which are representatives for dicotyledonous.

The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 12.8, 42.2, 139, 459, 1515, and 5000 mg test item/kg dry mass corresponding to 2.50, 8.23, 27.1, 89.5, 295, and 975 mg a.s./kg dry mass, respectively. A concentration of 1000 mg/kg corresponds to the limit test concentration recommended by OECD-guideline 208. The test item was incorporated into the test soil at the beginning of the experiment, and the effects on seedling emergence and growth were determined 14 days after emergence of 50 % of the control seedlings. Five replicates were prepared for the control and per concentration.

There was no concentration dependent effect on seedling emergence of P. aureus (mung bean) and R. sativus (radish) up to and including 5000 mg test item per kg dry soil. The EC50 was found to be > 5000 mg/kg dry soil. Also, there were no statistically significant effects on seedling emergence due to the test item. Thus, the NOEC for the species tested was found to be ≥ 5000 mg test item /kg dry soil corresponding to 975 mg a.s./kg dry soil.

There was no concentration dependent effect on growth (on fresh weight basis) of P. aureus up to and including 5000 mg test item per kg dry soil. The EC50 was found to be > 5000 mg/kg dry soil. Also, there were no statistically significant effects on growth of this species due to the test item. Thus, the NOEC for P. aureus was found to be ≥ 5000 mg test item /kg dry soil corresponding to 975 mg a.s./kg dry soil.

There was no concentration dependent effect on seedling survival of P. aureus and R. sativus up to and including 5000 mg test item per kg dry soil corresponding to 975 mg a.s./kg dry soil. The EC50 was found to be > 5000 mg test item/kg dry soil. Also, there were no statistically significant effects on seedling emergence due to the test item. Thus, the NOEC for the species tested was found to be ≥ 5000 mg/kg dry soil corresponding to 975 mg a.s./kg dry soil.

However, Raphanus sativus showed effects starting at concentrations above 1515 mg test item /kg dry soil. The EC50 was found to be 4109 mg test item per kg dry soil (95 % CL: 3321 – 5434) corresponding to 801

mg a.s./kg dry soil. The NOEC for Raphanus sativus was found to be 1515 mg test item/kg dry soil corresponding to 295 mg a.s./kg dry soil.

According to these results, the test item had a mild inhibitory effect on growth in one of the two plant species tested. With respect to seedling emergence and post-emergence survival, no inhibitory effect occurred.

Reference 3

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-10-22 to 2015-01-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: DIN EN ISO 22030

Version / remarks:

2011

Deviations:

yes

Remarks:

Deviating from the guideline, substrate for oat cultivation was not moistened to 80 % WHC due to the soil properties. From experience we know that moisture exceeding 60 % WHC significantly results in seed putrefaction in the soil applied.

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Düsseldorf, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 313, 625, 1250, 2500 and 5000 mg/kg soil dw (test start), 313, 1250 and 5000 mg/kg soil dw (test end), triplicate samples

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil: The volume of a stock solution (21.0 g in 100 mL) required to obtain the desired concentrations in the soil was filled up to 100 mL with water (except of the 50 mL sample) and added to 2159 g fresh mass soil (representing 2100 g dry mass soil). Additional 105.4 mL of water - used for purging and moistening – were added to the soil (now adjusted on approx. 35 % water holding capacity). 3100 g fresh mass soil was moistened with 223 mL water to adjust on ca. 35 % WHC for control soil. The soil was mixed thoroughly and placed into the test container.
- Controls: The control consisted of test substrate only. Six replicates were conducted. However, due to technical reasons (glass fibre wick was disconnected with the water reservoir) one replicate of the B. rapa controls dried out during emergence period and had to be discarded. For evaluation of effects on B. rapa only five replicates were available. However, the requirement of the guideline regarding a minimum number of four replicates is still fulfilled.

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oat
- Plant family: Poaceae
- Source of seed: Raiffeisen Genossenschaft Nordwest eG, Germany, May 2011, Cultivar, charge: no information provided
- Seed storage: The seeds were stored in the dark at the test facility at room temperature (20 ± 5 °C) and protected from moisture until use.

Species:

Brassica rapa

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: turnip
- Plant family: Brassicaceae
- Source of seed: Carolina Biological Supply Company, USA, Carolina® Wisconsin Fast Plants, Cultivar “Rapid-cycling Brassica rapa Rbr”, charge 15-8805
- Seed storage: The seeds were stored in the dark at the test facility at room temperature (20 ± 5 °C) and protected from moisture until use.

Test type:

other: growth and fertility test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

no

Total exposure duration:

47 d

Remarks:

68 days after apllication for Avena sativa, 50 days after application for Brassica rapa

Test temperature:

20 – 22 °C

pH:

4.9

Moisture:

Soil was only moistened to 35 % WHCmax for seed incorporation. However, soil was moistened to 55 % WHCmax subsequently for germination.

Details on test conditions:

TEST SYSTEM
- Test container (type, material, size): Plastic pots, approximately 9 x 9 cm side length and with a soil surface area of ca. 81 cm².
- Amount of soil: ca. 530 g moist test substrate (ca. 500 g dry soil).
- Method of seeding: Seeds were sown according to agricultural practice and the recommended seeding depths for the species.
- No. of seeds per container: 10
- No. of plants (retained after thinning): 8
- No. of replicates per treatment group: 4
- No. of replicates per control: 6 (for B. rapa only five replicates were available.)

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
The soil used in the test was a sieved (4 - 5 mm) natural sandy soil (Certified RefeSol 01-A; batch IME-04, loamy sand [DIN], Org C: 1.09 %, pH 4.9, sand: 77 %, silt: 17 %, clay: 6 % [DIN], WHCmax: 291 g/kg, CECeff: 11.9 mmolc/kg). The soil was not sterilized.

NUTRIENT MEDIUM (if used)
- Description: Continuous bottom watering of the test container via glass fibre wicks. The water was amended with fertilizer as necessary.
COMPO Grünpflanzendünger was used as fertilizer. Manufactory: COMPO GmbH & Co. KG, Münster, Germany. The concentration applied in the test was 1 mL fertilizer per liter water.
Nutrient content of COMPO Grünpflanzendünger
Ammonium nitrogen 37 g/L Iron (Chelate) 0.50 g/L
Nitrate nitrogen 33 g/L Copper (Chelate) 0.02 g/L
Phosphate 30 g/L Manganese (Chelate) 0.20 g/L
Potassium oxide 60 g/L Molybdenum 0.01 g/L
Magnesium oxide 60 g/L Zinc (Chelate) 0.02 g/L
Sulfur 18 g/L Boron 0.10 g/L

GROWTH CONDITIONS
- Photoperiod: 16 hours per day
- Light intensity and quality: 11,400 – 14,800 lx
- Day/night temperatures: 20 – 22 °C
- Relative humidity (%): 60 – 85 %
- Watering regime and schedules: The watering took place from the bottom from a reservoir according to consumption. A glass fibre wick, originating from a water reservoir and passing through the bottom of the container, was used to ensure consistent soil moisture.

EFFECT PARAMETERS MEASURED:
- Growth phase according to BBCH scale
- Total number of flowers (oat)
- Number of seed pods with fertile seeds (turnip)
- Fresh mass and length of shoots (without inflorescences or seed pods)
- Fresh mass of inflorescences (oat) and seed pods (turnip)
- Water content and dry weight of shoots, inflorescences and seed pods per vessel
- Mortality as a percentage of the number of plants after lens out at day 7 after emergence.

TEST CONCENTRATIONS
The initial measured test item concentrations in the soil at test start (sampling directly after test item incorporation) corresponded to 93.7 %, 82.1 %, 81.2 %, 72.7 %, 77.9 %, and 75.4 % of the nominal dose introduced into the test vessels in the treatments with 156.3, 312.5, 625, 1250, 2500 and 5000 mg test item per kg dry soil nominal concentration, respectively.

Nominal and measured concentrations:

Nominal concentrations: 313 (tested with B. rapa only), 625, 1250, 2500 and 5000 mg test item per kg dry mass soil, representing 60.9 (tested with B. rapa only), 122, 244, 488 and 975 mg a.s./kg

Reference substance (positive control):

yes

Remarks:

trichloroacetate; A test with the reference substance is regularly performed at the test facility once a year, thus fulfilling guideline recommendations.

Key result

Species:

Avena sativa

Duration:

68 d

Dose descriptor:

NOEC

Effect conc.:

244 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Key result

Species:

Avena sativa

Duration:

68 d

Dose descriptor:

NOEC

Effect conc.:

244 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: fertility

Species:

Brassica rapa

Duration:

50 d

Dose descriptor:

NOEC

Effect conc.:

488 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Species:

Brassica rapa

Duration:

50 d

Dose descriptor:

NOEC

Effect conc.:

488 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: fertility

Species:

Brassica rapa

Duration:

50 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Avena sativa

Duration:

68 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

seedling emergence

Species:

Brassica rapa

Duration:

50 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Species:

Avena sativa

Duration:

68 d

Dose descriptor:

NOEC

Effect conc.:

>= 975 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival

Details on results:

- Phatological symptoms: None observed.
- Seedling emergence: The test item had no effects on seedling emergence of Brassica rapa and Avena sativa. The NOEC for emergence at test end was ≥ 5000 mg test item per kg dry soil, representing ≥ 975 mg a.s. per kg dry soil.

- Post-emergence survival: Brassica rapa and Avena sativa was not affected by the test item through the whole study. The NOEC for post-emergence survival at test end was ≥ 5000 mg test item per kg dry soil, representing ≥ 975 mg a.s. per kg dry soil.

- Shoot length and biomass: Brassica rapa and Avena sativa were affected significantly by the test item test item on growth day 14. At test end, the data on shoot length of Brassica rapa and Avena sativa still showed concentration depending effects. The NOEC for shoot length of Brassica rapa and Avena sativa at test end are 2500 and 1250 mg test item per kg dry soil, representing 488 and 244 mg a.s. per kg dry soil, respectively. Biomass of Avena sativa was still affected at test end.. Biomass of Brassica rapa was not affected anymore at test end. The NOEC for biomass of Brassica rapa and Avena sativa at test end are ≥ 5000 and 1250 mg test item per kg dry soil, representing ≥ 975 and 244 mg a.s. per kg dry soil, respectively.

- Fertility: Number of flowers in B. rapa were decreased significantly. The NOEC was found to be 2500 mg test item per kg dry soil, representing 488 mg a.s. per kg dry soil. Number of fertile seed pods at test end was significantly affected by the test item. The NOEC was found to be 2500 mg test item per kg dry soil, representing 488 mg a.s. per kg dry soil. Weight of fertile seed pods was not affected up to and including a concentration of 5000 test item per kg dry soil, representing 975 mg a.s. per kg dry soil, which was the highest concentration tested. All endpoints showed only a slight concentration depending response. Number of inflorescences in A. sativa was decreased significantly. The NOEC was found to be 1250 mg test item per kg dry soil, representing 244 mg a.s. per kg dry soil. Weight of inflorescences was not affected up to and including a concentration of 5000 test item per kg dry soil, representing 975 mg a.s. per kg dry soil, which was the highest concentration tested.

Results with reference substance (positive control):

In order to confirm the reproducibility of the study as well as the germination capacity and sensitivity of the test species, non-GLP Terrestrial Plants, Growth Tests with trichloroacetate were performed. The acute results (harvest at growth day 14) of the last reference study are in agreement with experience EC50-values obtained in this institute.

Seedling emergence:
Avena sativa: Control 88.3 % 100 mg TCA/kg 87.5 %
Brassica rapa: Control 100.0 % 100 mg TCA/kg 100.0 %
EC50 values could not be computed.

Post-emergence survival:
Avena sativa: Control 100.0 % 100 mg TCA/kg 93.8 %
Brassica rapa: Control 100.0 % 100 mg TCA/kg 93.8 %
EC50 values could not be computed.

EC50 for growth (shoot length):
Avena sativa: 89 mg/kg
Brassica rapa: > 100 mg/kg

NOEC for growth (shoot length):
Avena sativa: 10 mg/kg
Brassica rapa: 10 mg/kg

EC50 for biomass (fresh weight):
Avena sativa: 81 mg/kg
Brassica rapa: > 100 mg/kg

NOEC for biomass (fresh weight):
Avena sativa: 10 mg/kg
Brassica rapa: ≥ 100 mg/kg

Reported statistics and error estimates:

All statistical tests (depending on normal distribution and homogeneity of variances the Fisher`s Exact Binomial Test, Williams Multiple Sequential t-test, Dunnett`s Multiple t-test, or Welch t-test for the NOEC calculation; ECx calculation) were performed with the computer software ToxRat Professional by ToxRat® Solutions GmbH. For the ECx calculation a probitanalysis assuming log-normal distribution of the values was applied. Statistical comparisons were applied one-sided smaller (p ≤ 0.05) in accordance with the guideline, since the aim of the study was to obtain relevant chronic NOECs.

Validity criteria fulfilled:

yes

Remarks:

The test is considered valid since control show: A percent emergence of ≥ 75 % (mean of the replicates); A healthy growth and the occurrence of flowers at least after 3 weeks (fast growing turnip) and 8 weeks (oat); A maximum of one dead plant per vessel.

Conclusions:

The relevant chronic endpoint was number of inflorescences in A. sativa with a NOEC of 1250 mg tes item per kg dry soil, representing 244 mg a.s. per kg dry soil.

Executive summary:

The effect of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid on the emergence, growth and fertility of terrestrial plant seedlings Avena sativa (oat) and Brassica rapa (turnip) was investigated according to the guideline DIN EN ISO 22030.

The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 313, 625, 1250, 2500 and 5000 mg test item/kg dry soil, representing 60.9, 122, 244, 488 and 975 mg a.s./kg. For Brassica rapa, an additional treatment of 156 mg test item/kg dry soil (30.5 mg a.s./kg) was applied. The test item was incorporated into the test soil at the beginning of the experiment. The effects on seedling emergence, growth and fertility were determined after the plants have built inflorescences (oat) or even seed pods (turnip). Additionally, growth was determined 14 days after emergence of 50 % of the control seedlings, representing the acute endpoint according to OECD 208. Six and four replicates were prepared for the control and per concentration, respectively.

According to the guideline, evaluation was done on the basis of nominal test item concentrations. However, to verify test item concentration in the test matrix, start and end concentrations in the artificial soil were measured. Seedling emergence and post-emergence survival of Brassica rapa and Avena sativa was not affected up to and including a concentration of 5000 mg test item per kg dry soil, representing 975 mg a.s./kg, which was the highest concentration tested.

Growth measured as shoot length and biomass, was affected significantly by the test item TEGO on growth day 14 and still at test end for both test species (except of biomass of B. rapa at test end). The most sensitive NOEC for shoot growth (both, shoot length and fresh weight) of Brassica rapa and Avena sativa at test end were 2500 and 1250 mg test item per kg dry soil, representing 488 and 244 mg a.s./kg, respectively.

Fertility was affected significantly by the test item. The NOEC for number of inflorescences in A. sativa at test end was found to be 1250 mg test item per kg dry soil, representing 244 mg a.s. per kg dry soil. For B. rapa, the number of flowers at day 14 was significantly affected starting at 5000 mg test item per kg dry soil, representing 975 mg a.s. per kg dry soil. At test end number of seed pods was still affected. The NOEC for number of inflorescences in A. sativa at test end was found to be 1250 mg test item per kg dry soil, representing 244 mg a.s. per kg dry soil. However, the weight of the seed pods was not affected. Therefore, the relevant NOEC for B. rapa was found to be 2500 mg test item per kg dry soil, representing 488 mg a.s. per kg dry soil.

Description of key information

EC50 (14 d) = 363 mg a.i./kg soil dw (nominal, Lactuca sativa, OECD guideline 208, growth)

NOEC (68 d) = 244 mg a.i./kg soil dw (nominal, Avena sativa, DIN EN ISO 22030, growth)

Key value for chemical safety assessment

Short-term EC50 or LC50 for terrestrial plants:

363 mg/kg soil dw

Long-term EC10, LC10 or NOEC for terrestrial plants:

244 mg/kg soil dw

Additional information

Two short-term and one long-term study are available testing the toxicity of Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid to terrestrial plants.

The key short-term study assessed the effects on the emergence and growth of terrestrial plant seedlings according to OECD guideline 208 for 14 days. The following test species were used: Avena sativa (oat), Lactuca sativa (lettuce), Phaseolus aureus (mung bean), Raphanus sativus (radish), Sinapis alba (mustard), and Triticum aestivum (wheat). The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 0, 1.64, 4.10, 10.2, 25.6, 64, 160, 400, and 1000 mg a.i./kg dry mass, respectively. There was no concentration dependent effect on seedling emergence of Avena sativa, Phaseolus aureus, Raphanus sativus, and Triticum aestivum up to 1000 mg a.i./kg, the highest concentration tested. Thus, the NOEC (14 d) for these tested species was found to be ≥ 1000 mg a.i./kg dry soil. The EC50 (14 d) was found to be > 1000 mg a.i./kg dry soil. There was a concentration dependent effect on seedling emergence of Lactuca sativa and Sinapis alba starting at 400 mg a.i./kg (NOEC = 160 mg a.i./kg). However, the EC50 (14 d) for Lactuca sativa was found to be 1062 mg a.i./kg (by extrapolation), and that for Sinapis alba > 1000 mg a.i./kg dry soil. There were no statistically significant effects on growth (on a fresh weight per plant basis) of Triticum aestivum up to the highest test concentration (NOEC ≥ 1000 mg a.i./kg, EC50 > 1000 mg a.i./kg). Concentration related inhibition of plant growth was observed in Avena sativa (NOEC = 400 mg a.i./kg), Phaseolus aureus (NOEC ≤ 1.64 mg a.i./kg), Raphanus sativus (NOEC = 10.2 mg a.i./kg), Lactuca sativa (NOEC = 160 mg a.i./kg), and Sinapis alba (NOEC = 400 mg a.i./kg). However, for the first three of these five species, the effects were only moderate, with the growth rate never being inhibited by more than 50 %. Accordingly, the EC50 (14 d) values for these tested species were found to be > 1000 mg a.i./kg dry soil. For Lactuca sativa and Sinapis alba, EC50 (14 d) values of 363 mg a.i./kg and 400–1000 mg a.i./kg, respectively, were found.

Another short-term toxicity test tested the effects on Amines, N-C10-C16-alkyltrimethylenedi-, reaction products with chloroacetic acid according to the OECD guideline 208. The following test species were used: Phaseolus aureus (mung bean) and Raphanus sativus (radish).The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 12.8, 42.2, 139, 459, 1515, and 5000 mg test item/kg dry mass corresponding to 2.50, 8.23, 27.1, 89.5, 295, and 975 mg a.i./kg dry mass, respectively. The NOEC (14 d) for P. aureus was found to be ≥ 5000 mg/kg dry soil corresponding to 975 mg a.i./kg dry soil. The relevant NOEC (14 d) for the most sensitive endpoint (growth of Raphanus sativus) was found to be 1515 mg test item per kg dry soil, representing 295 mg a.i./kg. The EC50 (14 d) was calculated to be 4109 mg test item per kg dry soil, representing 801 mg a.i./kg. According to these results, the test item had a mild inhibitory effect on growth in one of the two plant species tested. With respect to seedling emergence and post-emergence survival, no inhibitory effect occurred.

In the long term-study assessed the effects on the emergence, growth and fertility of terrestrial plant seedlings A. sativa and Brassica rapa (turnip) according to the guideline DIN EN ISO 22030 for 68 and 50 days, respectively.

The seeds were placed in a natural sandy soil containing the test item at nominal concentrations of 313, 625, 1250, 2500 and 5000 mg test item/kg dry soil, representing 60.9, 122, 244, 488 and 975 mg a.i./kg. For B. rapa, an additional treatment of 156 mg test item/kg dry soil (30.5 mg a.i./kg) was applied. The effects on seedling emergence, growth and fertility were determined after the plants have built inflorescences or even seed pods. Additionally, growth was determined 14 days after emergence of 50 % of the control seedlings, representing the acute endpoint according to OECD 208.

According to the guideline, evaluation was done on the basis of nominal test item concentrations. Seedling emergence and post-emergence survival of B. rapa and A. sativa was not affected up to and including a concentration of 5000 mg test item per kg dry soil, representing 975 mg a.i./kg, which was the highest concentration tested.

Fertility was affected significantly by the test item. The NOEC (68 d) for number of inflorescences in A. sativa at test end was found to be 1250 mg test item per kg dry soil, representing 244 mg a.i. per kg dry soil. For B. rapa, the number of flowers at day 14 was significantly affected starting at 5000 mg test item per kg dry soil, representing 975 mg a.i. per kg dry soil. At test end number of seed pods was still affected. The NOEC (50 d) for number of inflorescences in A. sativa at test end was found to be 1250 mg test item per kg dry soil, representing 244 mg a.i. per kg dry soil. However, the weight of the seed pods was not affected. Therefore, the relevant NOEC (50 d) for B. rapa was found to be 2500 mg test item per kg dry soil, representing 488 mg a.i. per kg dry soil.

The most sensitive NOEC for shoot growth (both, shoot length and fresh weight) of B. rapa and A. sativa at test end were 2500 and 1250 mg test item per kg dry soil, representing 488 and 244 mg a.i./kg, respectively.