Registration Dossier

Administrative data

Endpoint:
toxicity to terrestrial plants: short-term (with study design considered suitable for long-term assessment)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
This study was conducted between 02 December 2016 and 02 May 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Relability 1 is assigned as the study was conducted to OECD TG 208 without deviation that would affect the study outcome and in compliance with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 208 (Terrestrial Plants, Growth Test)
Version / remarks:
19 July 2006
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Sponsor's identification: Oxirane, Mono (C12-14-alkyloxy) methyl derives
Description : clear colourless liquid
Batch number : AG 94A572900
Date received : 01 March 2010
Expiry date : 31 August 2015
Storage conditions: room temperature in the dark
Specific details on test material used for the study:
Chemical name: Oxirane, mono[(C12-14-alkyloxy)methyl] derivs
Product name : ARALDITE® DY-E /CH
CAS No .: 68609-97-2
Chemical formula: C4-H10-O.x(C3-H6-O.C2-H4-O)x.xl2
Purity/Concentration: Complex mixture used as product 100% (no C of A available)
Batch number : AAD1326100
Expiration date : 29 August 2019
Appearance : Clear liquid
Storage conditions: Refrigerated (2 - 8°C)

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Chemical analysis was carried out to verify the concentration of Oxirane in the acetone solutions prepared at 222.4, 2.746 and 0.915 mg/mL (applications of 1000, 12.34 and 4.12 mg/kg dry soil respectively). Aliquots (2 x 3 mL) of the treatment solutions were supplied in glass vials. Two 3 ml aliquots of prepared solutions at 222.4 and 2.746 mg/mL for the additional lettuce application were also analysed.

Test substrate

Vehicle:
yes
Remarks:
Acetone
Details on preparation and application of test substrate:
Test Soil
The soil used in the study was a sandy loam, batch number GIL1401. The percentage organic matter was 1.89%, by calculation, based on the measured organic carbon percentage of 1.1%. The carbon and potassium contents, pH and particle size of the soils were analysed and the soil was sieved prior to use to remove all particles >2 mm.

Range finding
A preliminary range finder phase was carried out at test concentrations of 0.1, 1, 10, 100 and 1000 mg/kg dry soil. For cabbage, carrot and lettuce five replicate pots of three seeds were sown, with eight pots of two seeds for tomato and four pots of four seeds for oat and onion.
Based on the results of the range finding, a dose response test was conducted at rates of 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species with an additional application at 4.12 mg/kg dry soil for tomato.

Test Item Preparation and Analysis
Treatments and soil preparation
Prior to treatment the moisture content of the soil was determined to be 17.53% and the appropriate amount of moist soil for each treatment was weighed out. Aliquots of sand were also weighed out for each treatment rate, for application of the test substance in solvent. Details are presented in the table overleaf.

Seven treatments were applied to all species in the order of untreated water control, solvent control and Oxirane at 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil. An additional application was made for tomato at 4.12 mg/kg dry soil.

Method of preparation and application
The application rates for cabbage, carrot, tomato, oat and onion were prepared in acetone as shown in the table below.

Application rate
(mg/kg dry soil) Volume of test substance (mL) Final volume of acetone (mL) Concentration (mg/mL) Solution code
1000 125 500 222.4 A
333.33 166.67 of A 500 74.133 B
111.11 166.67 of B 500 24.711 C
37.04 166.67 of C 500 8.237 D
12.34 166.67 of D 500 2.746 E
4.12 166.67 of E 500 0.915 F

The application for lettuce was repeated as germination in the controls did not meet the guideline criteria on the first occasion.
Lettuce only

Application rate
(mg/kg dry soil) Volume of test substance (mL) Final volume of acetone (mL) Concentration (mg/mL) Solution code
1000 25 100 222.4 A
333.33 33.33 of A 100 74.133 B
111.11 33.33 of B 100 24.711 C
37.04 33.33 of C 100 8.237 D
12.34 33.33 of D 100 2.746 E

The solvent solutions were applied to dry sand and when the acetone had evaporated, the sand was mixed into the appropriate amount of weighed moist soil. This soil was then moistened with reverse osmosis (RO) water and mixed using a handheld mixer to ensure that the soil had a structure suitable for sowing the seeds before it was distributed among labelled replicate pots.

Species Weight of moist soil (g) Weight of sand (g) Total weight of dry substrate (g) Volume of solvent solution applied (mL) Volume of RO water (mL)
Cabbage + carrot 17312.0 270 15,000 67.44 200
Lettuce 8886.4 139 7,700 34.62 103
Tomato 12695.6 198 11,000 49.465 147
Oat + onion 12695.6 198 11,000 49.456 147

The treated soil was then distributed among labelled replicate pots. For the untreated water and solvent control groups the same amounts of sand and moist soil was used as for the treatment groups. For the water control the dry sand was mixed into the moist soil and for the
solvent controls the appropriate volume of acetone was added and allowed to evaporate off. Water was then added as for the treated groups. The pH of the soil was recorded after application see table below.

Plant species Solvent control Water control (mg/kg dry soil)
4.12 12.34 37.04 111.11 333.33 1000
Cabbage + carrot 6.9 7.0 N/a 7.0 7.0 6.8 6.8 7.0
Lettuce 7.0 7.0 N/a 7.0 7.0 7.0 6.9 6.9
Tomato 7.0 6.9 7.1 7.2 7.1 6.9 6.9 6.9
Oat + onion 7.0 7.0 N/a 7.1 7.1 7.0 6.9 6.9
N/a not applicable.



Test organisms

open allclose all
Species:
Brassica oleracea var. capitata
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.
Species:
Daucus carota
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.
Species:
Lactuca sativa
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.
Species:
Lycopersicon esculentum
Plant group:
Dicotyledonae (dicots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.
Species:
Avena sativa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.
Species:
Allium cepa
Plant group:
Monocotyledonae (monocots)
Details on test organisms:
Species were selected based on recommendations in the OECD guidelines.

The seeds were purchased from commercial suppliers. Full details of the suppliers are recorded in the raw data. Seeds were stored dry in glass bottles and kept in darkness prior to the start of the study. Germination trials indicated that seed viability was > 70% for all species.

Study design

Test type:
seedling emergence and seedling growth test
Study type:
laboratory study
Substrate type:
natural soil
Remarks:
sandy loam, batch number GIL1401
Limit test:
no
Remarks:
Harvested 14-17 days after germination

Test conditions

Test temperature:
19.1 - 26.2”C
Moisture:
Relative humidity = 38 - 94%
Details on test conditions:
Exposure conditions
Experimental design
A total of 14 replicates, each containing three seeds were used for cabbage, carrot and lettuce with 20 replicates of two seeds for tomato and ten replicates of four seeds for oat and onion. For all species 9.5 cm internal diameter plastic pots were used, each filled to a depth of approximately 75 mm. The seeds were spaced out on the soil surface and approximately 10 mm of the same soil was added to cover the seeds. Each pot was labeled with the study number, species name, dose group, replicate number, treatment date and the name of the Study Director. Each application rate was also identified by a colour coded plastic stick label.

Environmental conditions
Plants were watered with reverse osmosis water as required, maintaining seedling health and growth (visually determined on a per-species and per-treatment basis).

Plants were maintained in controlled environment rooms providing a continuous 16-hour period of light per day. Temperatures were maintained at levels suitable for the individual species. Temperature and humidity were recorded continuously using the Rees system. Light intensity, (initially at soil level then at the top of the canopy after germination) was recorded daily at five random positions for each species throughout the exposure period. A summary of environmental data is given below.

Plantspecies Environmental parameters
Air temperature (°C) Relative humidity (%) Light (lux)
Cabbage 19.1 – 24.3 59.33 – 93.95 4170 - 8830
Carrot 19.1 – 24.3 59.33 – 93.95 5190 - 8440
Lettuce 19.3 – 26.2 38.07 – 85.10 5470 - 8940
Tomato 19.1 – 24.3 59.33 – 93.95 3540 - 8940
Oat 19.1 – 24.3 59.33 – 93.95 4420 - 17670
Onion 19.1 – 24.3 59.33 – 93.95 3310 - 9340
Nominal and measured concentrations:
Analysis of the concentrations of Oxirane in the treatment solutions confirmed nominal levels of the test substance were within applied limits ± 20% for all of the samples analysed, confirming accurate formulation
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Species:
Brassica oleracea var. capitata
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
943.26 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Lactuca sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Lycopersicon esculentum
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Allium cepa
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Brassica oleracea var. capitata
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
111.11 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
111.11 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Lactuca sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Lycopersicon esculentum
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Allium cepa
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
12.34 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Height
Species:
Brassica oleracea var. capitata
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
847.92 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Lactuca sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
958.39 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Lycopersicon esculentum
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
980.57 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Allium cepa
Duration:
22 d
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Brassica oleracea var. capitata
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Lactuca sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Lycopersicon esculentum
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Avena sativa
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
333.33 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Daucus carota
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
37.04 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
Allium cepa
Duration:
22 d
Dose descriptor:
NOEC
Effect conc.:
111.11 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: Dry Weight
Species:
other: All Species Tested
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Daucus carota
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Lactuca sativa
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Lycopersicon esculentum
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Avena sativa
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Allium cepa
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Species:
Brassica oleracea var. capitata
Dose descriptor:
NOEC
Effect conc.:
333.3 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
germination
Remarks on result:
other: exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion
Details on results:
Range finding
A range finding test was initially performed at rates of 0.1, 1, 10, 100 and 1000 mg/kg dry soil.
There were reductions in germination of 6.7% for cabbage at 1000 mg/kg dry soil and 20, 6.7 and 33.3% at 1, 100 and 1000 mg/kg dry soil respectively for lettuce with 40% plant mortality recorded at 1000 mg/kg dry soil.

Plant growth inhibition was observed in cabbage and lettuce at rates of 10, 100 and
1000 mg/kg dry soil and carrot, tomato, oat and onion at 1000 mg/kg dry soil only.

There were statistically significant reductions in height at 1000 mg/kg dry soil for all species.
There were statistically significant reductions in dry weight for cabbage, carrot, lettuce and onion at 1000 mg/kg dry soil, tomato at 10, 100 and 1000 mg/kg dry soil and oat at 100 and 1000 mg/kg dry soil.

Based on these results a dose response test was conducted at rates of 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species, with an additional application at 4.12 mg/kg dry soil for tomato.

Definitive
Formulation analysis
Analysis of the concentrations of Oxirane in the treatment solutions confirmed nominal levels of the test substance were within applied limits ± 20% for all of the samples analysed, confirming accurate formulation.

Germination and survival
There were no statistically significant effects on germination or survival in any plant species at any treatment rate except for cabbage at 1000 mg/kg dry soil where 17% reduction in germination in comparison to the solvent control (p<0.05) was recorded.
Germination in the water control was 100% for cabbage, carrot, lettuce, tomato and oat and 92.5% for onion. Germination in the solvent control was 100% for carrot, lettuce, tomato and oat and 97.6 and 97.5% for cabbage and onion respectively.
EC50 values for germination and survival were estimated to be >1000 mg/kg dry soil for all plant species.
The NOEC values for effects on germination and survival were 1000 mg/kg dry soil for all plant species

Effects on plant growth and health
The visual assessment, based on the overall condition of surviving seedlings for each replicate, was conducted at the end of the study and at regular intervals throughout. This assessment was conducted using a uniform scoring system based on plant size and vigour where 0 indicated no abnormal effects and 10 indicated all plants within a replicate being dead.
No signs of phytotoxicity were observed in the control groups.
The predominant treatment-related effect in surviving plants was growth inhibition.
At harvest, measurement of the height and dry weight for each surviving seedling from each replicate pot was conducted. the Results are summarised in Table 2 (below).
Following statistical analysis significant reductions in height in comparison to the solvent control were calculated for cabbage and carrot at 333.33 and 1000 mg/kg dry soil; for lettuce, tomato and oat, at 1000 mg/kg dry soil and for onion at 37.04, 111.11, 333.33 and 1000 mg/kg dry soil.
EC25 values for effects on height were 448.66, 976.36 and 462.37 mg/kg dry soil for cabbage, lettuce and onion respectively and >1000 mg/kg dry soil for carrot, tomato and oat.
EC50 values for effects on height were 943.26 mg/kg dry soil for cabbage and >1000 mg/kg dry soil for carrot, lettuce, tomato, oat and onion.
The NOEC values for effects on height were 111.11 mg/kg dry soil for cabbage and carrot, 333.33 mg/kg dry soil for lettuce, tomato and oat and 12.34 mg/kg dry soil for onion
Significant reductions in dry weight in comparison to the solvent control were calculated for cabbage, lettuce, tomato and oat at 1000 mg/kg dry soil; for carrot at 111.11, 333.33 and 1000 mg/kg dry soil and onion at 333.33 and 1000 mg/kg dry soil.
EC25 values for effects on dry weight were 518.42, 632.39, 874.55, 894.78, 717.65 and 806.87 mg/kg dry soil for cabbage, carrot, lettuce, tomato, oat and onion respectively.
EC50 values for effects on dry weight were 847.92, 958.39 and 980.57 mg/kg dry soil cabbage, lettuce and tomato respectively and >1000 mg/kg dry soil for carrot, oat, onion.
The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage, lettuce, tomato and oat, 37.04 mg/kg dry soil for carrot and 111.11 mg/kg dry soil for onion.
The study was considered valid as the validation criteria were met and no visible treatment related effects were observed in the controls.

Any other information on results incl. tables

Table 2: Summary of effects on height and dry weight

Species

Treatment

Application rate

(mg/kg dry soil)

Geometric mean height (cm)

% increase (+) or reduction (-) in mean height

pvalue

Geometric mean dry weight (mg)

% increase (+) or reduction (-) in mean dry weight

pvalue

Cabbage

Water control

0

4.425

+11

0.251T

28.005

+22

0.135T

Solvent control

0

3.995

-

-

22.884

-

-

Oxirane

12.34

4.072

+2

1.000W

26.516

+16

1.000W

37.04

4.014

+0.5

1.000W

25.082

+10

1.000W

111.11

3.839

-4

0.831W

28.546

+25

1.000W

333.33

3.146

-21

0.009**W

21.111

-8

0.727W

1000

1.904

-52

<0.001***W

10.526

-54

<0.001***W

Carrot

Water control

0

 2.313

+3

0.534T

 4.360

-3

0.781T

Solvent control

0

 2.240

-

-

 4.474

-

-

Oxirane

12.34

 2.188

-2

1.000W

 3.802

-15

0.189W

37.04

 2.372

+6

1.000W

 4.049

-10

0.189W

111.11

 2.034

-9

0.077W

 3.672

-18

0.042*W

333.33

 1.921

-14

0.004**W

 3.513

-21

0.012*W

1000

 1.761

-21

<0.001***W

 3.199

-29

<0.001***W

Lettuce

Water control

0

1.106

+0.5

0.932T

9.403

+42

0.002**T

Solvent control

0

1.101

-

-

6.641

-

-

Oxirane

12.34

1.233

+12

0.172D

9.927

+49

0.002**D

37.04

1.219

+11

0.252D

10.519

+58

<0.001***D

111.11

1.305

+19

0.015*D

9.557

+44

0.007**D

333.33

1.247

+13

0.114D

10.449

+57

<0.001***D

1000

0.840

-24

<0.001***D

3.470

-48

<0.001***D

pvalues are for the comparison with the solvent control using Williams' test(W)Dunnett’s test(D)andt-test(T)

*p<0.05, **p<0.01, ***p<0.001

- not applicable

Summary of effects on height and dry weight (continued)

 

Species

Treatment

Application rate

(mg/kg dry soil)

Geometric mean height (cm)

% increase (+) or reduction (-) in mean height

pvalue

Geometric mean dry weight (mg)

% increase (+) or reduction (-) in mean dry weight

pvalue

Tomato

Water control

0

 5.205

+3

0.494T

50.501

-1

0.920T

Solvent control

0

 5.031

 -

-

50.994

-

-

Oxirane

 

4.12

 4.683

-7

0.505D

 45.450

-11

0.692D

12.34

 4.825

-4

0.905D

 50.378

-1

1.000D

37.04

 5.215

+4

0.950D

 56.934

+12

0.728D

111.11

 5.142

+2

0.996D

 54.316

+7

0.969D

333.33

 5.213

+4

0.953D

 52.617

+3

1.000D

 

1000

 3.848

-24

<0.001***D

 22.825

-55

<0.001***D

Oat

Water control

0

39.482

+2

0.530T

61.353

+3

0.708T

Solvent control

0

38.763

0

-

59.779

-

-

Oxirane

 

12.34

39.439

+2

1.000W

57.111

-4

1.000W

37.04

38.993

+1

1.000W

60.084

+0.5

1.000W

111.11

38.096

-2

0.824W

59.724

0

1.000W

333.33

38.329

-1

0.824W

56.557

-5

0.563W

1000

30.852

-20

<0.001***W

32.863

-45

<0.001***W

Onion

Water control

0

10.390

-5

0.331T

4.391

+3

0.656T

Solvent control

0

10.945

0

-

4.255

-

-

Oxirane

 

12.34

11.172

+2

1.000W

4.622

+9

1.000W

37.04

9.420

-14

0.007**W

3.927

-8

0.503W

111.11

9.107

-17

0.001**W

4.087

- 4

0.503W

333.33

8.354

-24

<0.001***W

3.641

-14

0.036*W

1000

7.850

-28

<0.001***W

3.134

-26

<0.001***W

                                                                          

pvalues are for the comparison with the solvent control using Williams' test(W), Dunnett’s test(D)and the

t-test(T)

 *p<0.05, **p<0.01 , ***p<0.001

- not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
There were no statistically significant effects on germination or survival in any plant species at any treatment rate except for cabbage at 1000 mg/kg dry soil where 17% reduction in germination in comparison to the solvent control (p<0.05) was recorded.
Germination in the water control was 100% for cabbage, carrot, lettuce, tomato and oat and 92.5% for onion. Germination in the solvent control was 100% for carrot, lettuce, tomato and oat and 97.6 and 97.5% for cabbage and onion respectively.
EC50 values for germination and survival were estimated to be >1000 mg/kg dry soil for all plant species.
The NOEC values for effects on germination and survival were 1000 mg/kg dry soil for all plant species.
Following statistical analysis significant reductions in height in comparison to the solvent control were calculated for cabbage and carrot at 333.33 and 1000 mg/kg dry soil; for lettuce, tomato and oat, at 1000 mg/kg dry soil and for onion at 37.04, 111.11, 333.33 and 1000 mg/kg dry soil.
EC25 values for effects on height were 448.66, 976.36 and 462.37 mg/kg dry soil for cabbage, lettuce and onion respectively and >1000 mg/kg dry soil for carrot, tomato and oat.
EC50 values for effects on height were 943.26 mg/kg dry soil for cabbage and >1000 mg/kg dry soil for carrot, lettuce, tomato, oat and onion.
The NOEC values for effects on height were 111.11 mg/kg dry soil for cabbage and carrot, 333.33 mg/kg dry soil for lettuce, tomato and oat and 12.34 mg/kg dry soil for onion
Significant reductions in dry weight in comparison to the solvent control were calculated for cabbage, lettuce, tomato and oat at 1000 mg/kg dry soil; for carrot at 111.11, 333.33 and 1000 mg/kg dry soil and onion at 333.33 and 1000 mg/kg dry soil.
EC25 values for effects on dry weight were 518.42, 632.39, 874.55, 894.78, 717.65 and 806.87 mg/kg dry soil for cabbage, carrot, lettuce, tomato, oat and onion respectively.
EC50 values for effects on dry weight were 847.92, 958.39 and 980.57 mg/kg dry soil for cabbage, lettuce and tomato respectively and >1000 mg/kg dry soil for carrot, oat, onion.
The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage, lettuce, tomato and oat and 37.04 mg/kg dry soil for carrot and 111.11 mg/kg dry soil for onion.
The study was considered valid as the validation criteria were met and no visible phytotoxic effects were observed in the controls.
Executive summary:

This study was carried out with cabbage, carrot, lettuce, tomato, oat and onion to determine if the test substance exerts a detrimental effect to plants during early critical stages in their development in a maximum dose test designed to evaluate the phytotoxic effects of the test substance.

Following initial range finding at 0.1, 1, 10, 100 and 1000 mg/kg, the definitive study was conducted as a dose response test at 12.34, 37.04, 111.11, 333.33 and 1000 mg/kg dry soil for all species with an additional rate of 4.12 mg/kg dry soil for tomato.  

Each plant species was sown into treated soil.  A total of 14 pots of three seeds each were sown for cabbage, carrot and lettuce, 20 pots of two seeds each for tomato and 10 pots of four seeds each for oat and onion for all application rates of Oxirane, mono[(C12-14-alkyloxy) methyl] derivs, (due to the length of the name the test item will be referred to as Oxirane throughout the remainder of the report) and the water and solvent controls.  The soil used was a sandy loam, sieved to remove all particles > 2mm.

After application and sowing, the pots were checked daily and the numbers of emerged seedlings were recorded.  Emergence, survival and sub-lethal effects were recorded daily until at least 50% emergence of control seedlings and thereafter every one to three days.  Plants were harvested 14 - 17 days after 50% germination in the controls, giving a total exposure period of 21 days for cabbage, lettuce and oat and 22 days for carrot, tomato and onion. The height of each individual plant was measured and they were then dried before being weighed to obtain a measurement of dry weight.

Effect on germination and plant survival

There were no statistically significant effects on germination or survival in any plant species at any treatment rate except for cabbage at 1000 mg/kg dry soil where 17% reduction in germination in comparison to the solvent control (p<0.05) was recorded.  

Germination in the water control was 100% for cabbage, carrot, lettuce, tomato and oat and 92.5% for onion.  Germination in the solvent control was 100% for carrot, lettuce, tomato and oat and 97.6 and 97.5% for cabbage and onion respectively.

EC50 values for germination and survival were estimated to be >1000 mg/kg dry soil for all plant species.

The NOEC values for effects on germination and survival were 1000 mg/kg dry soil for all plant species.

Effect on plant growth and health

No signs of phytotoxicity were observed in the control groups.

The predominant treatment-related effect in surviving plants was growth inhibition.

At harvest, measurement of the height and dry weight for each surviving seedling from each replicate pot was conducted.  

Following statistical analysis significant reductions in height in comparison to the solvent control were calculated for cabbage and carrot at 333.33 and 1000 mg/kg dry soil; for lettuce, tomato and oat at 1000 mg/kg dry soil and for onion at 37.04, 111.11, 333.33 and 1000 mg/kg dry soil.

EC25 values for effects on height were 448.66, 976.36 and 462.37 mg/kg dry soil for cabbage, lettuce and onion respectively and >1000 mg/kg dry soil for carrot, tomato and oat.  

EC50 values for effects on height were 943.26 mg/kg dry soil for cabbage and >1000 mg/kg dry soil for carrot, lettuce, tomato, oat and onion.

The NOEC values for effects on height were 111.11 mg/kg dry soil for cabbage and carrot, 333.33 mg/kg dry soil for lettuce, tomato and oat and 12.34 mg/kg dry soil for onion

Significant reductions in dry weight in comparison to the solvent control were calculated for cabbage, lettuce, tomato and oat at 1000 mg/kg dry soil; for carrot at 111.11, 333.33 and 1000 mg/kg dry soil and onion at 333.33 and 1000 mg/kg dry soil.

EC25 values for effects on dry weight were 518.42, 632.39, 874.55, 894.78, 717.65 and 806.87 mg/kg dry soil for cabbage, carrot, lettuce, tomato, oat and onion respectively.

EC50 values for effects on dry weight were 847.92, 958.39 and 980.57 mg/kg dry soil for cabbage, lettuce and tomato respectively and >1000 mg/kg dry soil for carrot, oat, onion.

The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage, lettuce, tomato and oat and 37.04 mg/kg dry soil for carrot and 111.11 mg/kg dry soil for onion.

Conclusion

There were no statistically significant effects on germination or survival in any plant species at any treatment rate except for cabbage at 1000 mg/kg dry soil where 17% reduction in germination in comparison to the solvent control (p<0.05) was recorded. 

Germination in the water control was 100% for cabbage, carrot, lettuce, tomato and oat and 92.5% for onion. Germination in the solvent control was 100% for carrot, lettuce, tomato and oat and 97.6 and 97.5% for cabbage and onion respectively.

EC50values for germination and survival were estimated to be >1000 mg/kg dry soil for all plant species.

The NOEC values for effects on germination and survival were 1000 mg/kg dry soil for all plant species.

Following statistical analysis significant reductions in height in comparison to the solvent control were calculated for cabbage and carrot at 333.33 and 1000 mg/kg dry soil; for lettuce, tomato and oat, at 1000 mg/kg dry soil and for onion at 37.04, 111.11, 333.33 and 1000 mg/kg dry soil.

EC25values for effects on height were 448.66, 976.36 and 462.37 mg/kg dry soil for cabbage, lettuce and onion respectively and >1000 mg/kg dry soil for carrot, tomato and oat. 

EC50values for effects on height were 943.26 mg/kg dry soil for cabbage and >1000 mg/kg dry soil for carrot, lettuce, tomato, oat and onion.

The NOEC values for effects on height were 111.11 mg/kg dry soil for cabbage and carrot, 333.33 mg/kg dry soil for lettuce, tomato and oat and 12.34 mg/kg dry soil for onion

Significant reductions in dry weight in comparison to the solvent control were calculated for cabbage, lettuce, tomato and oat at 1000 mg/kg dry soil; for carrot at 111.11, 333.33 and 1000 mg/kg dry soil and onion at 333.33 and 1000 mg/kg dry soil.

EC25values for effects on dry weight were 518.42, 632.39, 874.55, 894.78, 717.65 and 806.87 mg/kg dry soil for cabbage, carrot, lettuce, tomato, oat and onion respectively.

EC50values for effects on dry weight were 847.92, 958.39 and 980.57 mg/kg dry soil for cabbage, lettuce and tomato respectively and >1000 mg/kg dry soil for carrot, oat and onion.

The NOEC values for effects on dry weight were 333.33 mg/kg dry soil for cabbage, lettuce, tomato and oat and 37.04 mg/kg dry soil for carrot and 111.11 mg/kg dry soil for onion.

The study was considered valid as the validation criteria were met and no visible phytotoxic effects were observed in the controls.