Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 1 of 23

Data Requirement: PMRA Data Code: 9.8.4 (TGAI) or 9.8.6 (EP)

EPA DP Barcode: 416756

OECD Data Point: IIA 8.12 (TGAI) and IIIA 10.8.1.1 (EP)

EPA Guideline: 850.4100

Test material: Nicosulfuron 40 g/L SC Purity: 39.8%

Common name

Chemical name: IUPAC: 

CAS name: 2-[[[[(4,6-Dimethoxy-2-pyrimidinyl)amino]carbonyl]amino]sulfonyl]-N,N-dimethyl3-pyridinecarboxamide

CAS No.: 111991-09-4

Synonyms: DPX-V9360 40 g/L SC, DPX-V9360-155, DPX-V9360, Nicosulfuron 40 g/L SC, 3-

Pyridinecarboxamide, 2--[[[[(4,6-Dimethoxy-2-pyrimidinyl)amino]carbonyl]amino]sulfonyl]-N,NdimethylPrimary Reviewer: Dana Worcester Signature: 

Senior Scientist, CDM Smith Date: 12/17/14

Secondary Reviewer: Teri S. Myers Signature: 

Senior Scientist, CDM Smith Date: 04/03/15 

Primary Reviewer: Hannah Yingling Date: 9/18/2015

Biologist, EPA

CITATION: Porch, J.R. and K.H. Martin. Nicosulfuron (DPX-V9360) 40 G/L SC: A Greenhouse Study to Investigate 

the Effects on Seedling Emergence and Growth of Ten Terrestrial Plants Following Soil Exposure. Unpublished 

study performed by Wildlife International, Easton, Maryland and sponsored by E. I. du Pont de Nemours and 

Company, Wilmington, Delaware. DuPont Study Number: DuPont-21826. Study completed August 27, 2007.

DISCLAIMER: This document provides guidance for EPA and PMRA reviewers on how to complete a data 

evaluation record after reviewing a scientific study concerning the acute toxicity of a pesticide to terrestrial vascular 

plants. It is not intended to prescribe conditions to any external party for conducting this study nor to establish 

absolute criteria regarding the assessment of whether the study is scientifically sound and whether the study satisfies 

any applicable data requirements. Reviewers are expected to review and to determine for each study, on a case-bycase basis, whether it is scientifically sound and provides sufficient information to satisfy applicable data 

requirements. Studies that fail to meet any of the conditions may be accepted, if appropriate; similarly, studies that 

meet all of the conditions may be rejected, if appropriate. In sum, the reviewer is to take into account the totality of 

factors related to the test methodology and results in determining the acceptability of the study. 
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 2 of 23

EXECUTIVE SUMMARY:

The effect of Nicosulfuron 40 g/L SC on the seedling emergence of monocot (corn, Zea mays; oat, Avena sativa; 

onion, Allium cepa; and sorghum, Sorghum bicolor) and dicot (cucumber, Cucumis sativus; oilseed rape, Brassica 

napus; pea, Pisum sativum; soybean, Glycine max; sugarbeet Beta vulgaris; and tomato, Lycopersicon esculentum) 

crops was studied at a nominal concentration for oilseed rape of 0 (negative), 0.000024, 0.000073, 0.00022, 

0.00066, 0.0020, 0.0059, 0.018 and 0.053 lb ai/A; and 0.00042, 0.00083, 0.0017, 0.0034, 0.0067, 0.013, 0.027 and 

0.053 lb ai/A for all others. The measured concentration was 0.053 lb ai/A for corn, oat, onion, sorghum and oilseed 

rape; 0.052 lb ai/A for cucumber, pea, sugarbeet and tomato; and 0.056 lb ai/A for soybean. 

The growth medium used in the seedling emergence test was standard soil (loam, pH 6.2, organic carbon 1.0%). On 

day 21 the surviving plants per pot were recorded and cut at soil level for measuring the plant height and dry weight

The emergence in the negative control ranged from 83 to 95%. The reviewer found significant inhibitions in 

emergence for sugarbeet of 29% at 0.00083 lb ai/A treatment (p<0.05; Dunnett’s test). There were no significant 

effects at any other treatment level for sugarbeet. There were no significant effects on emergence for any other 

species. 

The reviewer measured survival based on number planted. Negative control survival based on number planted 

ranged from 20 to 100%. The reviewer found significant inhibitions in survival compared to the negative control for 

sugarbeet, with inhibition of 43% at the 0.052 lb ai/A treatment level (p<0.05; Dunnett’s test). There were no 

significant inhibitions in survival compared to the negative control for any other species. 

There were significant inhibitions in dry weight for all species except corn, oats and pea. Inhibitions compared to the 

negative control in cucumber dry weight of 35% at 0.052 lb ai/A; onion of 42% at the 0.053 lb ai/A; oilseed rape of 

36 and 65% at the 0.018 and 0.053 lb ai/A; sorghum of 20-68% beginning at the 0.0017 lb ai/A treatments; soybean 

of 22 and 38% at the 0.027 and 0.056 lb ai/A; sugarbeet of 33 and 72% at the 0.027 and 0.052 lb ai/A; and tomato of 

46% at the 0.052 lb ai/A (Williams test, p<0.05).

There were significant inhibitions in height for all species except corn, pea and tomato. The reviewer found 

significant decrease in height compared to the negative control for cucumber of 29% at the 0.052 lb ai/A treatment 

(Dunnetts test, p<0.05). There were also significant decrease in oats height compared to the negative control of 27, 

35 and 15% at the 0.013, 0.027 and 0.053 lb ai/A treatments; in onion of 23% at the 0.053 lb ai/A; in oilseed rape of 

15, 46 and 77% at the 0.0059, 0.018 and 0.053 lb ai/A levels; in sorghum of 19-71% beginning at the 0.0017 lb ai/A 

treatments; in soybean of 14, 10, and 22% at the 0.013, 0.027 and 0.056 lb ai/A; and in sugarbeet of 42% at the 

0.052 lb ai/A treatments (Williams test, p<0.05).

 

Based on the reviewers results, the most sensitive monocot species was sorghum based on dry weight, with NOAEC 

and IC25 values of 0.00083 and 0.000954 lb ai/A, respectively; and the most sensitive dicot species was oilseed rape 

based on height with NOAEC and IC25 values of 0.002 and 0.0089 lb ai/A, respectively. 

The occurrence of phytotoxic effects increased in severity and prevalence with increasing test concentrations; 

symptoms included wilting, stunting, leaf deformation, bleaching, chlorosis, necrosis, and mortality. The visual 

response score of 100 indicated normal seedlings and 0 indicated complete mortality. Maximum effects were 35-73 

for all species with the exception corn and tomato, which had maximum effects of 100 and 93, respectively. Effects 

were dose-related. 

Maximum Labeled Rate: 0.053 lb ai/A (equivalent to 60 g as/ha)
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 3 of 23

Results Synopsis

Most sensitive monocot: Sorghum, based on dry weight 

EC50/IC50: 0.00678 lb ai/A 95% C.I.: 0.00462-0.00996 lb ai/A

EC25/IC25: 0.000954 lb ai/A 95% C.I.: 0.000436-0.00185 lb ai/A

EC05/IC05: 0.0000567 lb ai/A 95% C.I.: N/A-0.000401 lb ai/A

NOEC: 0.00083 lb ai/A

Slope: N/A 95% C.I.: N/A

Most sensitive dicot: Oilseed rape, based on height

EC50/IC50: 0.0207 lb ai/A 95% C.I.: 0.0168-0.0255 lb ai/A

EC25/IC25: 0.0089 lb ai/A 95% C.I.: 0.00599-0.0123 lb ai/A

EC05/IC05: 0.00264 lb ai/A 95% C.I.: N/A-0.00441 lb ai/A

NOEC: 0.002 lb ai/A

Slope: N/A 95% C.I.: N/A

Table 1 (Tier II studies). Summary of most sensitive parameters by species (lbs ai/A).

Species Endpoint NOEC EC05/IC05 EC25/IC25 EC50/IC50

Corn Dry weight 0.053 0.000636 NC NC

Cucumber Dry weight 0.027 0.000012 0.0105 1.17

Oats Height 0.0067 0.000535 0.0329 0.577

Onion Height 0.027 0.00514 0.0535 0.273

Oilseed Rape Height 0.002 0.00264 0.0089 0.0207

Pea None 0.052 >0.052 >0.052 >0.052

Sorghum Dry weight 0.00083 0.0000567 0.000954 0.00678

Soybean Dry weight 0.013 0.00789 0.0418 0.133

Sugarbeet Dry weight 0.027 0.0118 0.0225 0.0354

Tomato Dry weight 0.027 0.0024 0.0179 0.0724

This study is scientifically sound and is classified as acceptable.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 4 of 23

I. MATERIALS AND METHODS

GUIDELINE FOLLOWED: This study was conducted in compliance with OPPTS 850.4100: Seedling 

emergence and seedling growth (January 2012) and OECD 208 Guideline 

for test of chemicals, Terrestrial Plant Test: Seedling Emergence and 

Seedling Growth Test (July 2006). The reviewer evaluated the study 

methods according to EPA Ecological Effects Test Guidelines, OCSPP 

Guideline 850.4100: Seedling Emergence and Seedling Growth. There 

were some deficiency and deviations noted by the reviewer.

1. The physico-chemical properties of the test material were not reported.

The deficiency and deviations did not have an impact on the acceptability of this study.

COMPLIANCE: Signed and dated GLP, Quality Assurance and Data Confidentiality 

statements were provided. This study was conducted in compliance with 

Good Laboratory Practice Standards as published by USEPA, August 17, 

1989, OECD Principle of Good Laboratory Practice (Paris, 1998); and

Japan MAFF, October 1, 1999. 

A. MATERIALS:

1. Test Material Nicosulfuron 

Description: Not reported.

Lot No./Batch No.: DPX-V9360-155

Purity: 38.7%

Stability of compound 

under test conditions: Analytical determinations based on measured concentration of the test 

material in the initial spray solution yielded a recoveries of 98-105% of 

nominal (n = 4). 

(OECD recommends chemical stability in water and light)

Storage conditions of 

test chemicals: The test material was stored at <40oC.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 5 of 23

Table 2. Physical/chemical properties of Nicosulfuron

Parameter Values Comments

Water solubility at 20 ̊C Not reported

Vapor pressure Not reported

UV absorption Not reported

pKa Not reported

Kow Not reported

2. Test organism: 

Monocotyledonous species: Oats (Avena sativa, Poaceae), corn (Zea mays, Poaceae), onion (Allium cepa, 

Lilaceae) and sorghum (Sorghum bicolar, Poaceae); EPA recommends four monocots in two families, 

including corn.

Dicotyledonous species: Cucumber (Cucumis sativus; Cucurbitaceae); Oilseed rape (Brassica napus, 

Brassicaceae); pea (Pisum sativum, Fabaceae); Soybean (Glycine max, Fabaceae); Sugar beet (Beta 

vulgaris; Chenopodiaceae); and Tomato (Lycopersicon esculentum, Solanaceae); EPA recommends six 

dicots in four families, including soybean and a root crop.

OECD recommends a minimum of three species selected for testing, at least one from each of the following categories: 

Category 1: ryegrass, rice, oat, wheat, and sorghum; Category 2: mustard, rape, radish, turnip, and Chinese cabbage; 

Category 3: vetch, mung bean, red clover, fenugreek, lettuce, and cress.

Seed source: Corn and oilseed rape from Pioneer Hi-Bred International, Inc.; Cucumber from Ferry-Morse 

Meyer Seed Co.; Oat from Gries Seed Farm; Onion from Wannamaker Seeds; Pea and Tomato from Meyer 

Seed Co.; Sorghum from N.C. Organics; Soybean from Missouri Foundation Seeds; and Sugarbeet from 

Betaseed Inc.

Prior seed treatment/sterilization: The seeds were not treated with any type of fungicides, insecticides, or 

any pesticides.

Historical % germination of seed: Corn, 96%; cucumber, 82%; oat, 97%; oilseed rape, 100%; onion, 

>80%; pea, 85%; soybean, 98%; sugarbeet, 90%; and tomato, 90%.

Seed storage, if any: Not reported.

B. STUDY DESIGN:

1. Experimental Conditions 

a. Limit test: None.

b. Range-finding study: None.

c. Definitive Study
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 6 of 23

Table 3: Experimental Parameters - Seedling Emergence.

Parameters Seedling Emergence

Details Remarks

Criteria

Duration of the test 21 days

Recommended test duration is 14-21 days.

OECD recommends that the test be terminated no 

sooner than 14 days after 50 percent of the control 

seedlings have emerged

Number of 

seeds/plants/species/ 

replicate

4 pots (replicates) with 10 seeds 

per pot.

Ten seeds per replicate should be used.

OECD recommends a minimum of five seeds planted

in each replicate within 24 hours of incorporation 

of the test substance. All seeds of each species for 

each test should be of the same size class. The seed 

should not be imbibed.

Number of replicates

Control:

Adjuvant control:

Treated:

4

N/A

4

Four replicates per dose should be used.

OECD recommends a minimum of four replicates 

per treatment
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 7 of 23

Parameters Seedling Emergence

Details Remarks

Criteria

Test concentrations (lb 

ai/A)

Nominal:

Measured: 

All species except oilseed rape:

0 (negative), 0.00042, 0.00083, 

0.0017, 0.0034, 0.0067, 0.013, 

0.027 and 0.053 lb ai/A; 

Oilseed rape:

0 (negative), 0.000024, 0.000073, 

0.00022, 0.00066, 0.0020, 0.0059, 

0.018 and 0.053 lb ai/A.

Corn, oat, onion, and sorghum:

0 (negative), 0.00042, 0.00083, 

0.0017, 0.0034, 0.0067, 0.013, 

0.027 and 0.053 lb ai/A for corn, 

oat, onion, and sorghum; 

Cucumber, pea, sugarbeet, and 

tomato:

0 (negative), 0 0.00042, 0.00083, 

0.0017, 0.0034, 0.0067, 0.013, 

0.027and 0.052 lb ai/A;

Oilseed rape:

0 (negative), 0.000024, 0.000073, 

0.00022, 0.00066, 0.0020, 0.0059, 

0.018 and 0.053 lb ai/A;

Soybean

0 (negative), 0.00042, 0.00083, 

0.0017, 0.0034, 0.0067, 0.013, 

0.027 and 0.056 lb ai/A .

The high dose concentration was measured, all 

others were nominal.

Five test concentrations should be used with a dose 

range of 2X or 3X progression

OECD recommends three concentrations, 

preferably with application rates equivalent to 0.0 

(control), 1.0, 10.0 and 100 mg substance per kg of 

oven-dried soil.

Method and interval of 

analytical verification

LOQ:

LOD:

Duplicate samples were taken 

from the negative control and the 

treatment groups for analysis via 

HPLC with Ultracarb C8column

with UV detection (240 nm).

Not reported.

Lowest analytical standard 

analyzed.

Adjuvant (type, percentage, 

if used)

N/A
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 8 of 23

Parameters Seedling Emergence

Details Remarks

Criteria

Test container (pot)

Size/Volume

Material: 

(glass/polystyrene)

Pots with diameter of 16 cm and 

12 cm depth.

Plastic

Non-porous containers should be used.

OECD recommends that non-porous plastic or 

glazed pot be used.

Growth facility Greenhouse

Method/depth of seeding A template was used to prepare 10 

holes and seeds introduced 

manually. Seeds were covered 

depressing the soil surface.

Test material application

Application time including 

the plant growth stage

Number of application 

Application interval

Method of application

After planting.

1

N/A- single application

The test material was applied 

using a linear cabinet track 

sprayer (DeVries Research, Teejet 

8001 EVSV nozzle, 40 psi and 

applied 41 cm from the soil 

surface.

Details of soil used

Geographic location

Depth of soil collection

Soil texture

% sand

N/A

N/A

Loam

51

Non-sterile top soil.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 9 of 23

Parameters Seedling Emergence

Details Remarks

Criteria

% silt

% clay

pH:

% organic carbon

CEC

Moisture at 1/3 atm (%)

38

11

6.2

1.0

7.1

14.6

Soil mixes containing sandy loam, loam, or clay 

loam soil with no greater than 2% organic matter 

are preferable. Glass beads, rock wool, and 100% 

acid washed sand are not preferred.

OECD prefers the soil to be sieved (0.5 cm) to 

remove coarse fragments. Carbon content should 

not exceed 1.5% (3% organic matter). Fine particles 

(under 20um) makeup should be between 10 and 

20%. The recommended pH is between 5.0 and 7.5.

Details of nutrient medium, 

if used

N/A

Watering regime and 

schedules

Water source/type:

Volume applied:

Interval of application:

Method of application:

Sub-irrigation.

Domestic well water.

Daily.

The plants were bottom watered

daily.

EPA prefers that bottom watering be utilized for 

seedling emergence studies so that the chemical is 

not leached out of the soil during the test.

Any pest control 

method/fertilization, if used

None reported.

Test conditions

Temperature:

Photoperiod:

Light intensity and quality:

Relative humidity:

16-31 ̊C for corn, oat, onion, 

sorghum and oilseed rape; 16-

30 ̊C for cucumber, pea, sugarbeet 

and tomato; 18-31oC for soybean

16L:8D

Natural sunlight supplemented 

with artificial light.

8.8-19.1 moles/m2/day for corn, 

oat, onion, sorghum and oilseed 

rape; 10.1-19.1 moles/m2/day for 

cucumber, pea, sugarbeet and 

tomato and 10.7-13.3 

moles/m2/day for soybean

13-86% for soybean and 9-80%

for all others

EPA prefers that the cold vs warm loving plants be 

tested in two separate groups to optimize plant 

growth.

OECD prefers that the temperature, humidity and 

light conditions be suitable for maintaining normal 

growth of each species for the test period.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 10 of 23

Parameters Seedling Emergence

Details Remarks

Criteria

Reference chemical (if 

used)

Name:

Concentrations: 

N/A

Other parameters, if any None

2. Observations: 

Table 4: Observation Parameters - Seedling Emergence.

Parameters Seedling Emergence

Details Remarks

Parameters measured (e.g., number 

of germinated seeds, emerged 

seedlings, plant height, dry weight 

or other endpoints)

- Emergence

- Survival

- Shoot height

- Total dry weight

- Phytotoxicity

Measurement technique for each 

parameter 

Emergence and 

phytotoxicity were 

visually determined. 

Survival was defined as 

the percent of emerged by 

the study author. Total

plant weight was 

estimated measuring the 

total dry weight per 

replicate following oven 

drying. 

Observation intervals Each pot was inspected 

weekly, emergence and 

phytotoxicity assessments 

performed. Dry weight 

and shoot height were 

recorded at study 

termination.

Other observations, if any N/A

Were raw data included? Yes
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 11 of 23

Phytotoxicity rating system, if used

0%= complete mortality or 

defoliation; 90%=at the threshold 

of visual detection and 100%=

No injury or effect.

II. RESULTS and DISCUSSION:

A. INHIBITORY EFFECTS:

1. Seedling Emergence: 

The emergence in the negative control ranged from 83 to 95%. The study author reported no significant 

inhibitions in emergence for any species at any treatment level (p>0.05). The reviewer found significant 

inhibitions in emergence for sugarbeet of 29% at 0.00083 lb ai/A treatment (p<0.05; Dunnett’s test). There 

were no significant effects at any other treatment level for sugarbeet. There were no significant effects on 

emergence for any other species. 

The study author reported % survived based on number emerged. The study author survival in the negative 

control ranged from 89 to 100%. The study author reported no significant inhibitions in survival for any 

species except sugarbeet, which had significant effects at the 0.052 lb ai/A (p<0.05; Jonckheere-Terpstra 

Step-Down test; inhibitions not reported). The reviewer measured survival based on number planted. 

Negative control survival based on number planted ranged from 20 to 100%. The reviewer found 

significant inhibitions in survival compared to the negative control for sugarbeet, with inhibition of 43% at

the 0.052 lb ai/A treatment level (p<0.05; Dunnett’s test). There were no significant inhibitions in survival 

compared to the negative control for any other species. 

The study author found inhibitions in dry weight for all species except corn, oats and pea. Cucumber had 

significant inhibitions compared to the negative control of up to a maximum of 35% beginning at 0.0067 lb 

ai/A; onion inhibitions of 42% at 0.053 lb ai/A; oilseed rape of up to 65% beginning at 0.0059 lb ai/A; 

sorghum of up to 68% beginning at 0.0017 lb ai/A, soybean of up to 38% beginning at 0.013 lb ai/A; 

sugarbeet of 72% at 0.053 lb ai/A and tomato of up to 46% beginning at 0.013 lb ai/A (p<0.05; JonckheereTerpstra Step-Down test). The reviewer found significant inhibitions in dry weight for all species except

corn, oat and pea. The reviewer found were significant inhibitions in dry weight for all species except corn, 

oats and pea. There were significant inhibitions compared to the negative control in cucumber dry weight of 

35% at 0.052 lb ai/A; onion of 42% at the 0.053 lb ai/A; oilseed rape of 36 and 65% at the 0.018 and 0.053 

lb ai/A; sorghum of 20-68% beginning at the 0.0017 lb ai/A treatments; soybean of 22 and 38% at the 0.027 

and 0.056 lb ai/A; sugarbeet of 33 and 72% at the 0.027 and 0.052 lb ai/A; and tomato of 46% at the 0.052 

lb ai/A (Williams test, p<0.05).

The study author found inhibitions in height for all species except corn, pea and tomato. Cucumber had 

significant inhibitions compared to the negative control of up to a maximum of 29% beginning at 0.013 lb 

ai/A; oat inhibitions of up to 35% beginning at 0.0067 lb ai/A; onion inhibitions of up to 23% at 0.0067 lb 

ai/A; oilseed rape of 46 and 77% at 0.018 and 0.053 lb ai/A; sorghum of up to 71% beginning at 0.0017 lb 

ai/A, soybean beginning at 0.013 lb ai/A (inhibitions not reported); and sugarbeet of 2 and 42% at 0.027 

and 0.052 lb ai/A (p<0.05; Jonckheere-Terpstra Step-Down test). The reviewer found significant 

inhibitions in height for all species except corn, pea and tomato. The reviewer found significant decrease in 

height compared to the negative control for cucumber of 29% at the 0.052 lb ai/A treatment (Dunnetts test, 

p<0.05). There were also significant decreases in oats height compared to the negative control of 27, 35 and 

15% at the 0.013, 0.027 and 0.053 lb ai/A treatments; in onion of 23% at the 0.053 lb ai/A; in oilseed rape 

of 15, 46 and 77% at the 0.0059, 0.018 and 0.053 lb ai/A levels; in sorghum of 19-71% beginning at the 
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 12 of 23

0.0017 lb ai/A treatments; in soybean of 14, 10, and 22% at the 0.013, 0.027 and 0.056 lb ai/A; and in

sugarbeet of 42% at the 0.052 lb ai/A treatments (Williams test, p<0.05).

Based on the study author’s results, the most sensitive monocot species was sorghum based on height, with 

NOAEC and ER50 values of 0.0008 and 0.0063 lb ai/A, respectively; and the most sensitive dicot species 

was oilseed rape based on height, with NOAEC and ER50 values of 0.0059 and 0.0204 lb ai/A, respectively.

The occurrence of phytotoxic effects increased in severity and prevalence with increasing test 

concentrations; symptoms included wilting, stunting, leaf deformation, bleaching, chlorosis, necrosis, and 

mortality. The visual response score of 100 indicated normal seedlings and 0 indicated complete mortality.

Maximum effects were 35-73 for all species with the exception corn and tomato, which had maximum 

effects of 100 and 93, respectively. Effects were dose-related. 

B. REPORTED STATISTICS: 

Emergence, survival, dry weight, and height means and standard deviations were determined for control and 

treatment groups. Treatments were compared to the negative control. NOEC was determined using JonckheereTerpstra test. EC25 and EC50 and their 95% confidence intervals were determined using non-linear regression 

analysis. All statistical determinations were made with 95% certainty. Nominal concentrations were used for all 

analyses.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

Plants: Seedling Emergence

PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 13 of 23

Table 5: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for height (lbs ai/A)*

height (cm) NOEC IC05 95%CI IC25 95%CI IC50 95%CI slope 95%CI

Corn 26.9-29 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber14.7-7.35 0.0067 ND N/A 0.0373 0.0016-

0.0533

>0.053 N/A N/A N/A

Oats214.8-23.1 0.0034 ND N/A 0.0329 0.0018-

0.6145

>0.053 N/A N/A N/A

Onion36.38-9.35 0.0034 ND N/A >0.53 N/A >0.053 N/A N/A N/A

Oilseed Rape41.73-8.3 0.0059 ND N/A 0.0088 0.0055-

0.0142

0.0204 0.0155-

0.0272

N/A N/A

Pea 8.8-11.1 >0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sorghum55.23-18 0.00083 ND N/A 0.0012 0.00042-

0.0082

0.0063 0.0019-

0.0214

N/A N/A

Soybean615.4-19.7 0.0067 ND N/A 0.0533 0.0000-

0.0000

>0.053 N/A N/A N/A

Sugarbeet73.23-6.73 0.0133 ND N/A 0.0401 0.0212-

>0.053

>0.053 N/A N/A N/A

Tomato 4.7-6.85 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

ND- Not determined. NC- Not calculable.

*All statistically significant inhibitions based on Jonckheere-Terpstra Step-Down test, p<0.05.

1 Significant inhibition in cucumber height of 21, 12 and 29% at the 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative control.

2 Significant inhibitions in oat height of 8, 27, 35 and 15% at the 0.0067, 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative 

control.

3 Significant inhibition in onion height of 2, 5, 7 and 23% at the 0.0067, 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative 

control.

4 Significant inhibition in oilseed rape height of 46 and 77% at the 0.018 and 0.053 lb ai/A levels compared to the negative control.

5 Significant inhibition in sorghum height of 18-71% beginning at the 0.0017 lb ai/A treatments compared to the negative control.

6 Significant inhibition in soybean height at the 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative control (inhibitions not 

reported).

7 Significant inhibition in sugarbeet of 2 and 42% at the 0.027 and 0.052 lb ai/A treatments compared to the negative control.
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Table 5a: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for biomass (lbs ai/A)*

weight (mg) NOEC IC05 95%CI IC25 95%CI IC50 95%CI slope 95%CI

Corn 1.39-1.67 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber10.675-1.06 0.0034 ND N/A 0.0106 0.00042-

>0.053

>0.53 N/A N/A N/A

Oats 0.429-0.606 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Onion20.0388-0.0773 0.0266 ND N/A 0.0281 0.0199-

0.0398

>0.053 N/A N/A N/A

Oilseed Rape30.0928-0.279 0.0020 ND N/A 0.0099 0.0050-

0.0198

0.0300 0.0202-

0.0445

N/A N/A

Pea 1.35-1.84 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sorghum40.168-0.519 0.0008 ND N/A 0.0010 0.0004-

0.0101

0.0068 0.0016-

0.0287

N/A N/A

Soybean52.32-3.75 0.0067 ND N/A 0.0328 0.0144-

>0.053

>0.053 N/A N/A N/A

Sugarbeet60.055-0.26 0.0266 ND N/A 0.0223 0.0072-

>0.053

0.0355 0.0186-

>0.053

N/A N/A

Tomato70.138-0.303 0.0067 ND N/A 0.0181 0.0052-

>0.053

>0.053 N/A N/A N/A

ND- Not determined. NC- Not calculable.

*All statistically significant inhibitions based on Jonckheere-Terpstra Step-Down test, p<0.05.

1 Significant inhibition in cucumber dry weight of 35, 28, 15 and 35% at the 0.0067, 0.13, 0.027 and 0.053 lb ai/A treatments compared to the 

negative control.

2 Significant inhibition in onion dry weight of 42% at the 0.053 lb ai/A treatments compared to the negative control.

3 Significant inhibition in oilseed rape dry weight of 22, 36 and 65% at the 0.0059, 0.018 and 0.053 lb ai/A levels compared to the negative control.

4 Significant inhibition in sorghum dry weight of 19-68% beginning at the 0.0017 lb ai/A treatments compared to the negative control.

5 Significant inhibition in soybean dry weight of 16, 22 and 38% at the 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative control.

6 Significant inhibition in sugarbeet dry weight of 72% at the 0.053 lb ai/A treatments compared to the negative control. 

7 Significant inhibition in tomato dry weight of 13, 13 and 46% at the 0.013, 0.027 and 0.052 lb ai/A treatments compared to the negative control.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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Page 15 of 23

Table 5b: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for emergence (lbs ai/A)

% NOEC EC05 95%CI EC25 95%CI EC50 95%CI slope 95%CI

Corn 88-98 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber 55-83 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Oats 83-95 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Onion 85-95 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Oilseed Rape 80-93 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Pea 88-98 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sorghum 93-100 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Soybean 85-98 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sugarbeet 68-90 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Tomato 75-90 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

ND- Not determined. NC- Not calculable.
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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Page 16 of 23

Table 5c: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for survival (lbs ai/A); based on # emerged

% NOEC EC05 95%CI EC25 95%CI EC50 95%CI Slope* 95%CI

Corn ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Oats ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Onion ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Oilseed Rape ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Pea ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sorghum ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Soybean ND 0.0563 ND N/A >0.053 N/A >0.053 N/A N/A N/A

Sugarbeet1 ND 0.053 ND N/A 0.044 0.036-

0.052

>0.053 N/A N/A N/A

Tomato ND 0.053 ND N/A >0.053 N/A >0.053 N/A N/A N/A

ND- Not determined. NC- Not calculable.

1 Significant inhibition at the 0.052 lb ai/A treatment compared to the negative control (Jonckheere-Terpstra Step-Down test, p<0.05; inhibition not 

reported).
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Page 17 of 23

Mid-study emergence

Control Corn Cucumber Oats Onion Oilseed 

Rape

Pea Sorghum Soybean Sugarbeet Tomato Formulation 

Blank

83-95 85-98 55-83 80-95 85-95 78-93 83-98 88-100 78-90 60-90 75-90 N/A

Plant Injury Index*

Control Corn Cucumber Oats Onion Oilseed 

Rape

Pea Sorghum Soybean Sugarbeet Tomato Formulation 

Blank

0 100 70-98 73-100 75-100 35-95 75-100 38-100 80-100 55-100 93-100 N/A

0%=Complete mortality; 100%=No visual response.

.

C. VERIFICATION OF STATISTICAL RESULTS BY THE REVIEWER:

All analyses were conducted comparing treated to the negative control. These analyses were conducted using 

CETIS version 1.8.7.12 and backend settings approved for use by EFED on 3/25/14. Data for each endpoint 

were tested to determine if their distributions were normal and if their variances were homogeneous using 

Shapiro-Wilk’s and Levene’s tests, respectively. Data that satisfied these assumptions were subjected to 

Dunnett’s and William’s tests, and data that did not satisfy these assumptions were subjected to the nonparametric MannWhitney-U and Jonckheere’s tests. Nominal concentrations were used for all analyses except 

for high dose concentrations which were measured. Linear (survival and emergence) and nonlinear (height and 

dry weight) regression models were used to interpret EC/ICx values
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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Table 6: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for height (lbs ai/A)

height 

(cm)

NOEC IC05 95%CI IC25 95%CI IC50 95%CI slope 95%CI

Corn 26.9-29 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber14.7-7.35 0.027 0.000585 N/A0.005160.0363 0.00844-

0.118

0.638 0.00669-

60.9

N/A N/A

Oats214.8-23.1 0.0067 0.000535 N/A0.002790.0329 0.0135-

0.072

0.577 0.0415-

8.02

N/A N/A

Onion36.38-9.35 0.027 0.00514 N/A0.01390.0535 0.0235-

0.104

0.273 0.0274-

2.73

N/A N/A

Oilseed Rape41.73-8.3 0.002 0.00264 N/A0.004410.0089 0.00599-

0.0123

0.0207 0.0168-

0.0255

N/A N/A

Pea 8.8-11.1 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Sorghum55.23-18 0.00083 0.000102 N/A0.0004310.0011

6

0.000674-

0.00186

0.00628 0.0047-

0.00839

N/A N/A

Soybean615.4-19.7 0.0067 0.0000707 N/A0.005052.2 0.000107-

683

>0.056 N/A N/A N/A

Sugarbeet73.23-6.73 0.027 0.00335 N/A0.01060.0231 0.0111-

0.0412

0.0882 0.0264-

0.294

N/A N/A

Tomato 4.7-6.85 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

ND- Not determined. NC- Not calculable.

1 Significant decrease in cucumber height, inhibition of 29% at the 0.052 lb ai/A treatments compared to the negative control (Dunnetts test, 

p<0.05).

2 Significant decrease in oats height, inhibition of 27, 35 and 15% at the 0.013, 0.027 and 0.053 lb ai/A treatments compared to the negative control 

(Williams test, p<0.05).

3 Significant decrease in onion height, inhibition of 23% at the 0.053 lb ai/A treatments compared to the negative control (Williams test, p<0.05).

4 Significant decrease in oilseed rape height, inhibitions of 15, 46 and 77% at the 0.0059, 0.018 and 0.053 lb ai/A levels compared to the negative 

control (Williams test, p<0.05).

5 Significant decrease in sorghum height, inhibition of 19-71% beginning at the 0.0017 lb ai/A treatments compared to the negative control 

(Williams test, p<0.05).

6 Significant decrease in soybean height, inhibition of 14, 10, and 22% at the 0.013, 0.027 and 0.056 lb ai/A treatments compared to the negative 

control (Williams test, p<0.05). 

7 Significant decrease in sugarbeet height, inhibition of 42% at the 0.052 lb ai/A treatments compared to the negative control (Williams test, 

p<0.05).
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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Table 6a: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for biomass (lbs ai/A)

weight (mg) NOEC IC05 95%CI IC25 95%CI IC50 95%CI slope 95%CI

Corn 1.39-1.67 0.053 0.000636 N/A0.00029NC N/A NC N/A N/A N/A

Cucumber10.675-1.06 0.027 0.000012 N/A0.003060.0105 0.00156-

0.0525

1.17 0.000936-

1470

N/A N/A

Oats 0.429-0.606 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Onion20.0388-0.0773 0.027 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Oilseed Rape30.0928-0.279 0.0059 0.00206 N/A0.004880.01 0.00499-

0.0171

0.0301 0.021-

0.0431

N/A N/A

Pea 1.35-1.84 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Sorghum40.168-0.519 0.00083 0.0000567 N/A0.0004010.000954 0.000436-

0.00185

0.00678 0.00462-

0.00996

N/A N/A

Soybean52.32-3.75 0.013 0.00789 N/A0.01930.0418 0.0239-

0.0659

0.133 0.0341-

0.518

N/A N/A

Sugarbeet60.055-0.26 0.013 0.0118 N/A0.02030.0225 0.008-

0.0346

0.0354 0.0264-

0.0473

N/A N/A

Tomato70.138-0.303 0.027 0.0024 N/A0.006780.0179 0.00964-

0.0299

0.0724 0.0318-

0.165

N/A N/A

ND- Not determined. NC- Not calculable.

1 Significant decrease in cucumber dry weight, inhibition of 35% at the 0.052 lb ai/A treatments compared to the negative control (Williams test, 

p<0.05).

2 Significant decrease in onion dry weight, inhibition of 42% at the 0.053 lb ai/A treatments compared to the negative control (Williams test, 

p<0.05).

3 Significant decrease in oilseed rape dry weight, inhibitions of 36 and 65% at the 0.018 and 0.053 lb ai/A levels compared to the negative control 

(Williams test, p<0.05).

4 Significant decrease in sorghum dry weight, inhibition of 20-68% beginning at the 0.0017 lb ai/A treatments compared to the negative control 

(Williams test, p<0.05).

5 Significant decrease in soybean dry weight, inhibition of 22 and 38% at the 0.027 and 0.056 lb ai/A treatments compared to the negative control 

(Williams test, p<0.05); 

6 Significant decrease in sugarbeet dry weight, inhibition of 33 and 72% at the 0.027 and 0.052 lb ai/A treatments compared to the negative control 

(Williams test, p<0.05).

7 Significant decrease in tomato dry weight, inhibition of 46% at the 0.052 lb ai/A treatments compared to the negative control Williams test, 

p<0.05).
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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PMRA Submission Number {..............} EPA MRID Number 49169704 

Page 20 of 23

Table 6b: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for emergence (lbs ai/A)

% NOEC EC05 95%CI EC25 95%CI EC50 95%CI slope 95%CI

Corn 88-98 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber 55-83 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Oats 83-95 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Onion 85-95 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Oilseed Rape 80-93 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Pea 85-98 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Sorghum 93-100 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Soybean 85-98 0.056 >0.056 N/A >0.056 N/A >0.056 N/A N/A N/A

Sugarbeet160-90 0.00042 N/C N/A N/C N/A N/C N/A N/A N/A

Tomato 83-90 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

ND- Not determined. NC- Not calculable.

1 There was a significant inhibition of 29% in sugarbeet dry weight at the 0.00083 lb ai/A level (Dunnett’s test, p<0.05).
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Table 6c: Effect of Nicosulfuron on 21-Day Seedling Emergence

Species Results summary for survival (lbs ai/A); based on # planted

% NOEC EC05 95%CI EC25 95%CI EC50 95%CI slope 95%CI

Corn 88-98 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Cucumber 50-78 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Oats 83-95 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Onion 85-95 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Oilseed Rape 80-88 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Pea 85-98 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

Sorghum 90-98 0.053 >0.053 N/A >0.053 N/A >0.053 N/A N/A N/A

Soybean 80-98 0.056 >0.056 N/A >0.056 N/A >0.056 N/A N/A N/A

Sugarbeet143-90 0.027 NC N/A NC N/A NC N/A N/A N/A

Tomato 93-90 0.052 >0.052 N/A >0.052 N/A >0.052 N/A N/A N/A

ND- Not determined. NC- Not calculable.

1 Significant decrease in sugarbeet survival, inhibition of 43% at the 0.052 lb ai/A treatments compared to the negative control (Dunnett’s test, 

p<0.05).
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Mid-study emergence

Control Corn Cucumber Oats Onion Oilseed 

Rape

Pea Sorghum Soybean Sugarbeet Tomato Formulation 

Blank

Not reported N/A

Plant Injury Index*

Control Corn Cucumber Oats Onion Oilseed 

Rape

Pea Sorghum Soybean Sugarbeet Tomato Formulation 

Blank

0 100 70-98 73-100 75-100 35-95 75-100 38-100 80-100 55-100 93-100 N/A

0%=Complete mortality; 100%=No visual response.

Monocot

Most sensitive monocot: Sorghum, based on dry weight

EC50/IC50: 0.00678 lb ai/A 95% C.I.: 0.00462-0.00996 lb ai/A

EC25/IC25: 0.000954 lb ai/A 95% C.I.: 0.000436-0.00185 lb ai/A

EC05/IC05: 0.0000567lb ai/A 95% C.I.: N/A-0.000401 lb ai/A

NOEC: 0.00083 lb ai/A

Slope: N/A 95% C.I.: N/A

Most sensitive dicot: Oilseed rape, based on height

EC50/IC50: 0.0207 lb ai/A 95% C.I.: 0.0168-0.0255 lb ai/A

EC25/IC25: 0.0089 lb ai/A 95% C.I.: 0.00599-0.0123 lb ai/A

EC05/IC05: 0.00264 lb ai/A 95% C.I.: N/A-0.00441 lb ai/A

NOEC: 0.002 lb ai/A

Slope: N/A 95% C.I.: N/A

D. STUDY DEFICIENCIES: 

1. The physico-chemical properties of the test material were not reported.

E. REVIEWER’S COMMENTS: 

The reviewer and study author results are in general agreement. The study author reported the most 

sensitive monocot species was sorghum based on height, with NOAEC and IR50 values of 0.0008 and 

0.0068 lb ai/A, respectively; and the most sensitive dicot species was oilseed rape based on height, with 

NOAEC and IR50 values of 0.0059 and 0.0155 lb ai/A, respectively. In comparison, based on the reviewers 
Data Evaluation Record on the Acute Toxicity of Nicosulfuron to Terrestrial Vascular 

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Page 23 of 23

results, the most sensitive monocot species was sorghum based on dry weight, with NOAEC and IC25 values 

of 0.00083 and 0.000954 lb ai/A, respectively; and the most sensitive dicot species was oilseed rape based 

on height, with NOAEC and IC25 values of 0.002 and 0.0089 lb ai/A, respectively. The study author based 

most sensitive species results on the EC50 value, while the reviewer based the most sensitive species on IC25. 

Also, if the study author had used their calculated IC25 for sorghum, dry weight would have been the most 

sensitive endpoint. These differences most likely resulted in the difference in the monocot endpoint of 

height vs. dry weight for sorghum. The reviewer’s results are presented in the Executive Summary and 

Conclusions sections of this DER.

The in-life portion of this study was initiated on March 13, 2007 to May 24, 2007. 

F. CONCLUSIONS: 

This study is scientifically sound and is classified as acceptable. The most sensitive monocot species was 

sorghum based on dry weight, with NOAEC and IC25 values of 0.00083and 0.000954 lb ai/A, respectively; 

and the most sensitive dicot species was oilseed rape based on height, with NOAEC and IC25 values of 

0.002 and 0.0089 lb ai/A, respectively.

Most sensitive monocot and IC25: Sorghum (dry weight, 0.000954 lb ai/A).

Most sensitive dicot and IC25: Oilseed rape (height, 0.0089 lb ai/A).

III. REFERENCES: 

1. U.S. Environmental Protection Agency, Series 850-Ecological Effects Test Guidelines (draft), PPPTS 

Number 850.4100: Terrestrial Plant Toxicity, Tier I (Seedling Emergence). 1996.

2. U.S. Environmental Protection Agency, Series 850-Ecological Effects Test Guidelines (draft), PPPTS 

Number 850.4225: Terrestrial Plant Toxicity, Tier II (Seedling Emergence). 1996.

3. Bruce, Robert D. and Donald J. Versteeg. 1992 A Statistical Procedure for Modeling Continuous 

Toxicity Data. Environmental Toxicology and Chemistry. 11:1485-1494.

4. U.S. Environmental Protection Agency, “Pesticide Assessment Guidelines, Subdivision J, Hazard 

Evaluation Nontarget Plants, Series 122-1 and 123-1”, 22 October 1982.

5. OECD, OECD: Guideline for the Testing of Chemicals, Guideline 208. Terrestrial Plant Test: 

Seedling Emergence and Seedling Growth Test, OECD, July 2006.

6. Frans, R. E. and Talbert, R. E., Design of Fields Experiment and the Measurements and Analysis of 

Plant Responses. Pages 15-23 in B. Truelove, ed. Research Methods in Weed Science. Southern Weed 

Science Society, Auburn University, Alabama, 1977.

7. SAS Institute, Inc. 1999. SAS/STAT User’s Guide, Version 8. Cary, NC, SAS Institute, Inc.