Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 1 of 24

Data Requirement: PMRA DATA CODE {............}

EPA DP Barcode 386906

OECD Data Point {............}

EPA MRID 48376401

EPA Guideline 850.4550

Test material: Nicosulfuron (DPX-V9360) 75WG plus dry surfactant (DPX-GFJ47)

Purity: Nicosulfuron 75WG contains 74.8% ai

Common name

Chemical name: IUPAC: not reported

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

 pyridinecarboxamide monohydrate (for nicosulfuron (DPX-V9360) active ingredient)

CAS No.: 111991-09-4 (for nicosulfuron active ingredient)

Synonyms: Nicosulfuron (DPX-V9360) 75WG plus Dry Surfactant (DPX-GFJ47) (1:6), 

Nicosulfuron 75WG plus DPX-GFJ47, DPX-V9360 75WG (Paste Extruded Granule), 

DPX-V9360-123 (Batch Number), DPX-V9360-122 (Batch Number), DPX-GFJ47-

001 (Batch Number), Nicosulfuron (DPX-V9360) 75WG, Nicosulfuron 75WG, 3-

Pyridinecarboxamide, 2-[[[[(4,6-dimethoxy-2-

pyrimidinyl)amino]carbonyl]amino]sulfonyl]-N,Ndimethyl, monohydrate (inverted 

CAS name for nicosulfuron active ingredient) DPX-GFJ47 (Paste Extruded Granule)

Primary Reviewer: Mia Howard Signature: 

Staff Scientist, Cambridge Environmental, Inc. Date: 10/05/12

Secondary Reviewer: Teri S. Myers Signature: 

Environmental Scientist, CDM Smith Date: 08/14/13 

Primary Reviewer: Geoffrey Sinclair, Biologisy Date: 8/26/13

EPA/OPP/EFED/ERB5

Secondary Reviewer(s): {......................................} Date: {............}

{EPA/OECD/PMRA}

Reference/Submission No.: {.....................}

Company Code {............} [For PMRA]

Active Code {............} [For PMRA] 

Use Site Category: {............} [For PMRA]

EPA PC Code 129008

Date Evaluation Completed: {dd-mm-yyyy}

CITATION: Sloman, T.L. 2004. Nicosulfuron (DPX-V9360) 75WG plus Dry Surfactant (DPX-GFJ47) (1:6): 

Influence on Growth and Growth rate of the Blue-Green Alga Anabaena flos-aquae. An unpublished study 

performed by E.I. du Pont de Nemours and Company, Haskell Laboratory for Health and Environmental 

Sciences, Newark, Delaware. Laboratory project ID: DuPont-13989. Study sponsored by E.I. du Pont de 
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 2 of 24

Nemours and Company, Wilmington, Delaware. Study initiated 06 February 2004 and completed 11 May 

2004.

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 aquatic 

nonvascular 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-by-case 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.

EXECUTIVE SUMMARY:

In a 96-hour acute toxicity study, cultures of blue-green alga (Anabaena flos-aquae) were exposed to Nicosulfuron 

75WG plus dry surfactant (DPX-GFJ47) at nominal total formulation concentrations of 0 (negative control), 43.75, 

87.5, 175, 350, and 700 mg/L under static conditions. Nominal Nicosulfuron 75WG concentrations were 0 (negative 

control), 6.25, 12.5, 25.0, 50, and 100 mg/L. Nominal DPX-GFJ47 surfactant concentrations were 0 (negative 

control), 37.5, 75, 150, 300, and 600 mg/L. Nominal nicosulfuron active ingredient concentrations were 0 (negative 

control), 4.68, 9.35, 18.7, 37.4, and 74.8 mg ai/L. Mean measured nicosulfuron active ingredient concentrations 

were <LOD (negative control), 4.95, 9.96, 20.15, 40, and 77.95 mg ai/L. The NOAEC and EC50/IC50 values based 

on cell density were <4.95 and 18 mg a.i/L, respectively. The NOAEC and EC50/IC50 values based on area under the 

curve were <4.95 and 18 mg a.i/L, respectively. The NOAEC and EC50/IC50 values based on growth rate were 

<4.95 and 39.91 mg a.i/L, respectively. The % growth inhibition of cell density in the treated algal culture as 

compared to the control ranged from 11 to 100%. The EC50 derived from cell density would classify nicosulfuron 

as slightly toxic to Anabaena flos-aquae.

No morphological abnormalities were noted.

This toxicity study is classified as scientifically sound or and satisfies the guideline requirement for an acute algal 

(Anabaena flos-aquae) toxicity study. It is therefore classified as acceptable.

Results Synopsis

Test Organism: Blue-green algae, Anabaena flos-aquae

Test Type (Flow-through, Static, Static Renewal): Static

Toxicity values based on mean measured concentrations of nicosulfuron:

Cell density

EC05: 7.7 mg ai/L 95% C.I.: 5.2 to 11 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 4.31 ± 0.541

Area under the curve

EC05: 6.6 mg ai/L 95% C.I.: 4.6 to 9.4 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 3.75 ± 0.365

Growth rate
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 3 of 24

EC05: Not calculable 95% C.I.: N/A

EC50: 39.91mg ai/L* 95% C.I.: 37.68 to 42.14 mg ai/L*

NOAEC: <4.95 mg ai/L**

Probit Slope: N/A

Endpoint(s) Effected: cell density, area under the growth curve, growth rate

*The EC50 and 95% confidence interval based on growth rate were not verified by the reviewer, due to a poor fit of 

the probit model. The values reported here are those calculated by the study author based on nominal nicosulfuron 

active ingredient concentrations.

**This value was based on the study author’s analyses. The study author originally reported the NOAEC based on 

growth rate as <4.68 mg ai/L, based on nominal nicosulfuron exposure concentrations. The reviewer converted the 

NOAEC to the equivalent mean measured nicosulfuron concentration.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 4 of 24

I. MATERIALS AND METHODS

GUIDELINE FOLLOWED: The study as conducted according to the US EPA OPPTS Ecological 

Effects Test Guidelines. The following deviations from the US EPA 

OCSPP Ecological Effects Test Guidelines number 850.4550 (2012) were 

noted:

1. Three replicates were used per level. A minimum of four replicates is required.

2. Test vessels were shaken on a shaking table at 105 rpm. It is recommended that test vessels be shaken 

by hand once or twice per day rather than continuously shaken.

3. The pH of the test medium was 6.28 to 7.41 at test initiation. It is recommended that the pH of the 

medium be 7.5 ± 0.1. Lower initial pH values were associated with higher test concentrations.

4. The storage conditions of the test substance were not reported.

5. The pH and total organic carbon, particulate matter, metal, pesticide, and chlorine contents of the 

dilution water were not reported.

6. The age of the algal inoculum at test initiation was not reported. A logarithmically growing culture, 

typically 3 to 7 days old, is recommended.

7. The concentrations of test substance were not low enough to determine a NOAEC value.

These deviations did not affect the validity of the study. While cyanobacteria may be influenced in changes in 

pH the trends observed in this study did not appear to be pH dependent. Three replicates provided sufficient 

statistical power to result treatment differences and thus arrive at appropriate endpoints and conclusions.

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

statements were provided. The study was conducted in compliance with the 

US EPA FIFRA (40 CFR part 160) GLP standards with one exception: no 

characterization was performed on the blend used in this study. However, 

Nicosulfuron 75WG was characterized.

A. MATERIALS:

1. Test material Nicosulfuron (DPX-V9360) 75WG plus dry surfactant (DPX-GFJ47)(1:6)

Description: Nicosulfuron 75WG is a light brown granule. The dry surfactant (DPXGFJ47) was a brown solid.

Lot No./Batch No. :

Purity: Nicosulfuron 75WG contains 74.8% active ingredient.

Stability of compound 

under test conditions: Stable. Day 4 measured nicosulfuron concentrations ranged from 105-

109% of day 0 measured concentrations.

(OECD recommends water solubility, stability in water and light, pKa, Pow, and vapor pressure of test 

compound)

Storage conditions of 

test chemicals: Not reported

Physicochemical properties of nicosulfuron.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 5 of 24

Parameter Values Comments

Water solubility at 20ΕC Not reported 99.9 mg 

nicosulfuron/L (132 

mg/L Nicosulfuron 

75 WG) in 20X 

AAP medium with 

785 mg/L DPXGFJ47 dry 

surfactant

Vapor pressure Not reported

UV absorption Not reported

pKa Not reported

Kow Not reported

2. Test organism: 

Name: Blue-green algae, Anabaena flos-aquae

EPA requires a nonvascular species: For tier I testing, only one species, S. capricornutum, to be 

tested; for tier II testing, S. costatum, A. flos-aquae, S. capricorntum, and a freshwater diatom is 

tested. 

OECD suggests the following species are considered suitable: S. capricornutum, S. subspicatus, and 

C. vulgaris. If other species are used, the strain should be reported

Strain: Not reported

Source: In-house cultures originally obtained from the Department of Botany, 

Culture Collection of Algae, The University of Austin, Austin, Texas

Age of inoculum: Not reported

Method of cultivation: Cultured and tested in AAP nutrient medium

B. STUDY DESIGN:

1. Experimental Conditions

a. Range-finding study. A 96-hour range-finding study was performed in which algae were exposed to 

a negative control and nominal concentrations of 0.001, 0.01, 0.1, 1, 10, and 100 mg ai/L (it was not 

specified if the concentrations were total formulation, Nicosulfuron 75WG, or active ingredient 

concentrations). Inhibitions of cell density relative to the negative control were -15, -26, -17, -9, -22, 

and 99% for concentrations of 0.001, 0.01, 0.1, 1, 10, and 100 mg ai/L, respectively.

b. Definitive Study

Table 1: Experimental Parameters
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 6 of 24

Parameter Details Remarks

Criteria

Acclimation period:

Culturing media and conditions: 

(same as test or not)

Health: (any mortality observed)

Continuously cultured in-house

Same as test

None reported

EPA recommends two week acclimation 

period.

OECD recommends an amount of algae 

suitable for the inoculation of test 

cultures and incubated under the 

conditions of the test and used when still 

exponentially growing, normally after 

an incubation period of about 3 days. 

When the algal cultures contain 

deformed or abnormal cells, they must 

be discarded.

Test system

Static/static renewal

Renewal rate for static renewal

Static

N/A

EPA expects the test concentrations to 

be renewed every 3 to 4 days (one 

renewal for the 7 day test, 3-4 renewals 

for the 14 day test). 

Incubation facility

Environmental chamber on a shaker 

table

Duration of the test 96 hours

EPA requires: 96-120 hours

OECD: 72 hours 

Test vessel

Material: (glass/stainless steel)

Size:

Fill volume:

Glass

250 mL

50 mL

Sterile Erlenmeyer flaks fitted with 

foam stoppers 

OECD recommends 250 ml conical 

flasks are suitable when the volume of 

the test solution is 100 ml or use a 

culturing apparatus.

Details of AAP nutrient medium

pH at test initiation: 6.28 to 7.41

The pH of the medium should be 7.5 

±0.1 at the beginning of the test.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 7 of 24

Parameter Details Remarks

Criteria

pH at test termination:

Chelator used:

Carbon source:

Salinity (for marine algae):

7.20 to 7.83

Na2EDTAx2H2O

NaHCO3

N/A

OECD recommends the medium pH 

after equilibration with air is ~8 

with less than .001 mmol/l of 

chelator if used.

EPA recommends 20X-AAP and 

chelating agents (e.g. EDTA) in the 

nutrient medium for optimum cell 

growth. Lower concentrations of 

chelating agents (down to one-third 

of the normal concentration 

recommended for AAP medium) may 

be used in the nutrient medium used 

for test solution preparation if it is

suspected that the chelator will 

interact with the test material. 

ASTM reference, E1415-91and D 

3978-80 (reapproved 1987).

If non-standard nutrient medium 

was used, detailed composition 

provided (Yes/No)

A standard medium was used and a 

detailed composition was also 

provided

Dilution water 

source/type:

pH:

salinity (for marine algae):

water pretreatment (if any):

Total Organic Carbon:

particulate matter:

metals:

pesticides:

chlorine:

Milli-Q (deionized) water

Not reported

N/A

Deionized

Not reported

Not reported

Not reported

Not reported

Not reported

EPA pH: Skeletonema costatum= ~8.0 

Others = ~7.5 from beginning to end of 

the test. EPA salinity: 30-35 ppt. EPA is 

against the use of dechlorinated water.

OECD: pH is measured at beginning of 

the test and at 72 hours, it should not 

normally deviate by more than one unit 

during the test.

Indicate how the test material is 

added to the medium (added 

directly or used stock solution)

Used a 700 mg formulation/L stock 

solution prepared in AAP nutrient 

medium.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 8 of 24

Parameter Details Remarks

Criteria

Aeration or agitation Shaken at 105 rpm OCSPP (2012) guidelines 

recommend that test vessels be 

shaken by hand once or twice per 

day rather than continuously shaken 

in order to prevent clumping of the 

cells.

Initial cells density 10,000 cells/mL

EPA requires an initial number of 3,000 

- 10,000 cells/mL. For Anabaena flosaquae, cell counts on day 2 are not 

required.

OECD recommends that the initial cell 

concentration be approximately 10,000 

cells/ml for S. capricornutum and S. 

subspicatus. When other species are 

used the biomass should be 

comparable.

Number of replicates

Control:

Solvent control:

Treatments:

3

N/A

3

OCSPP (2012) guidelines require a 

minimum of four replicates per level

EPA requires a negative and/or solvent 

control with 3 or more replicates per 

doses. Navicula sp.tests should be 

conducted with four replicate.

OECD preferably three replicates at 

each test concentration and ideally twice 

that number of controls. When a vehicle 

is used to solubilize the test substance, 

additional controls containing the 

vehicle at the highest concentration used 

in the test.

Test concentrations

Nominal total formulation:

Nominal Nicosulfuron 75WG:

0 (negative control), 43.75, 87.5, 

175, 350, and 700 mg/L

0 (negative control), 6.25, 12.5, 

*Mean measured nicosulfuron 

concentrations were calculated by 

the reviewer as the arithmetic mean 

of the measured nicosulfuron 

concentrations at days 0 and 4.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 9 of 24

Parameter Details Remarks

Criteria

Nominal DPX-GFJ47:

Nominal nicosulfuron active 

ingredient:

Mean measured* nicosulfuron 

active ingredient:

25.0, 50.0, and 100 mg/L

0 (negative control), 37.5, 75, 150, 

300, and 600 mg/L

0 (negative control), 4.68, 9.35, 

18.7, 37.4, and 74.8 mg ai/L

<0.0097 (<LOD, negative control), 

4.95, 9.96, 20.15, 40.00, and 77.95 

mg ai/L

EPA requires at least 5 test 

concentrations, with each at least 60% 

of the next higher one. 

OECD recommends at least five 

concentrations arranged in a geometric 

series, with the lowest concentration 

tested should have no observed effect on 

the growth of the algae. The highest 

concentration tested should inhibit 

growth by at least 50% relatively to the 

control and, preferably, stop growth 

completely. 

Solvent (type, percentage, if used) N/A

Method and interval of analytical 

verification

The concentration of nicosulfuron in 

the test solutions was measured on 

days 0 and 4 using high performance 

liquid chromatography

Test conditions 

Temperature:

Photoperiod:

Light intensity and quality:

24.8 to 25.0°C

Continuous

1830 to 2070 lux, cool-white 

fluorescent light

EPA temperature: Skeletonema: 20ΕC, 

Others: 24-25ΕC; EPA photoperiod: S. 

costatum 14 hr light/ 10 hr dark, 

Others: Continuous; EPA light: 

Anabaena: 2.0 Klux (±15%), Others: 4 -

5 Klux (±15%)

OECD recommended the temperature in 

the range of 21 to25oC maintained at ± 

2o

C and continuous uniform illumination 

provided at approximately 8000 Lux 

measured with a spherical collector.

Reference chemical (if used)

name:

concentrations:

N/A

Other parameters, if any N/A

2. Observations: 
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 10 of 24

Table 2: Observation parameters

Parameters Details Remarks

Criteria

Parameters measured including the 

growth inhibition/other toxicity 

symptoms

Healthy cell count

Area under the growth curve

Growth rate EPA recommends the growth of the algae 

expressed as the cell count per mL, biomass 

per volume, or degree of growth as 

determined by spectrophotometric means.

Measurement technique for cell 

density and other end points

Cell density was determined 

using a hemacytometer and 

compound microscope. The 

area under the growth curve was 

calculated based on cell density. 

The growth rate was calculated 

using a logarithmic equation 

based on cell density.

EPA recommends the measurement 

technique of cell counts or chlorophyll a

OECD recommends the electronic particle 

counter, microscope with counting 

chamber, fluorimeter, spectrophotometer, 

and colorimeter. (note: in order to provide 

useful measurements at low cell 

concentrations when using a 

spectrophotometer, it may be necessary to 

use cuvettes with a light path of at least 4 

cm).

Observation intervals Every 24 hours

EPA and OECD: every 24 hours.

Other observations, if any Cells were visually observed for 

abnormalities. A recovery study 

was performed subsequently to 

the definitive test in order to 

determine if the effect of the test 

substance was algistatic or 

algicidal.

Indicate whether there was an

exponential growth in the control

Yes. After 96 hours, the cell 

density of the negative control 

was 656.0x104 cells/mL. EPA requires control cell count at 

termination to be 2X initial count or by a 

factor of at least 16 during the test.

OECD: cell concentration in control 

cultures should have increased by a factor 

of at least 16 within three days.

Were raw data included? Yes

II. RESULTS and DISCUSSION:

A. INHIBITORY EFFECTS:

After 96 hours, the cell density of the negative control was 656.0x104 cells/mL, yielding inhibitions compared 

to the negative control of 11, 40, 52, 95, and 100% for nominal nicosulfuron active ingredient concentrations of 
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 11 of 24

4.86, 9.35, 18.7, 37.4, and 74.8 mg ai/L, respectively. The NOAEC and EC50 values calculated by the study 

author based on nominal nicosulfuron active ingredient concentrations were <4.68 and 14.77 mg ai/L, 

respectively.

After 96 hours, the growth rate of the negative control was 0.0676, yielding inhibitions compared to the 

negative control of 2, 8, 12, 46, and 100% for nominal nicosulfuron active ingredient concentrations of 4.86, 

9.35, 18.7, 37.4, and 74.8 mg ai/L, respectively. The NOAEC and EC50 values calculated by the study author 

based on nominal nicosulfuron active ingredient concentrations were <4.68 and 39.9 mg ai/L, respectively.

After 96 hours, the area under the curve of the negative control was 112.120x106

, yielding inhibitions compared 

to the negative control of 12, 47, 53, 92, and 100% for nominal nicosulfuron active ingredient concentrations of 

4.86, 9.35, 18.7, 37.4, and 74.8 mg ai/L, respectively. The NOAEC and EC50 values calculated by the study 

author based on nominal nicosulfuron active ingredient concentrations were <4.68 and 15.48 mg ai/L, 

respectively.

No morphological abnormalities were noted. There was an increase in pH during the study. From the results of 

the recovery test, the effects of the test substance were determined to be algistatic within 144 hours for nominal 

nicosulfuron concentrations <74.8 mg/L.

Table 3: Effect of Nicosulfuron 75WG plus dry surfactant (DPX-GFJ47) on algal growth (blue-green algae, 

Anabaena flos-aquae)

Treatment 

Total formulation 

(nominal 

Nicosulfuron 75 

WG) [nominal 

DPX-GFJ47 

surfactant] 

{nominal 

nicosulfuron active 

ingredient} <mean 

measured 

nicosulfuron active 

ingredient> 

concentrations 

mg/L

Initial cell

density (x104

cells/mL)

Cell density (x104 cells/mL) at

48 hours 72 hours 96 hours

cell count % inhibition

Negative control 1 26.7 109.3 656.0 N/A

43.75 (6.25) [37.5] 

{4.86} <4.95>

1 22.7 93.3 585.3 11

87.5 (12.5) [75] 

{9.35} <9.96>

1 14.7 89.3 390.6 40

175 (25.0) [150] 

{18.7} <20.15>

1 93.3 56.0 312.0 52
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 12 of 24

350 (50) [300] 

{37.4} <40.00>

1 8.0 14.7 34.7 95

700 (100) [600] 

{74.8} <77.95>

1 2.7 0 0 100

Reference chemical 

(if used)

N/A

Table 4: Effect of Nicosulfuron 75WG plus dry surfactant (DPX-GFJ47) on algal growth (blue-green algae, 

Anabaena flos-aqaue)

Treatment 

Total formulation 

(nominal 

Nicosulfuron 75 

WG) [nominal 

DPX-GFJ47 

surfactant] 

{nominal 

nicosulfuron active 

ingredient} <mean 

measured 

nicosulfuron active 

ingredient> 

concentrations 

mg/L

Growth rate 

(0-96 hour) 

% 

inhibition*

Area under 

the growth 

curve (x106

)

% 

inhibition*

Negative control 0.0676 N/A 112.120 N/A

43.75 (6.25) [37.5] 

{4.86} <4.95>

0.0664 2 98.680 12

87.5 (12.5) [75] 

{9.35} <9.96>

0.0622 8 71.120 37

175 (25.0) [150] 

{18.7} <20.15>

0.0598 12 53.000 53

350 (50) [300] 

{37.4} <40.00>

0.0365 46 9.480 92

700 (100) [600] 

{74.8} <77.95>

0.000 100 0.400 100

*Calculated by the reviewer relative to the control , using mean values

Table 5: Statistical endpoint values.* (calculated by the study author)

Statistical Endpoint Nominal total Nominal Nominal DPX- Nominal 
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 13 of 24

formulation 

concentration 

(mg/L) 

Nicosulfuron 75 

WG concentration 

(mg/L)

GFJ47 surfactant

(mg/L)

nicosulfuron active 

ingredient (mg 

ai/L)

Cell density

NOAEC

EC50 (95% C.I.)

<43.75

138.25 (115.78 to

160.72)

<6.25 

19.75 (16.54 to 

22.96)

<37.5

118.5 (99.24 to 

137.76)

<4.68

14.77 (12.37 to 

17.17)

Area under the growth curve

NOAEC

EC50 (95% C.I.)

<43.75

144.83 (127.61 to 

161.98)

<6.25

20.69 (18.23 to 

23.14)

<37.5

124.14 (109.38 to 

138.84)

<4.68

15.48 (13.64 to 

17.31)

Growth rate

NOAEC

EC50 (95% C.I.)

<43.75

373.52 (352.66 to 

394.38)

<6.25

53.36 (50.38 to 

56.34)

<37.5

320.16 (302.28 to 

338.04)

<4.68

39.91 (37.68 to 

42.14)

* Do not use this table, if the study was deemed unacceptable.

B. REPORTED STATISTICS:

The study author statistically analyzed the endpoints for cell density, area under the growth curve, and growth 

rate using nominal total formulation, Nicosulfuron 75WG, DPX-GFJ47, and nicosulfuron active ingredient 

concentrations. The EC50 values and 95% confidence intervals were determined using inverse weighted leastsquares non-linear regression. The EC50 value based on cell density was calculated using probit analysis. The 

NOAEC values based on cell density and area under the curve were determined using Jonckheere-Terpstra tests. 

The NOAEC value based on growth rate was determined using a non-parametric multiple comparison test.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 14 of 24

C. VERIFICATION OF STATISTICAL RESULTS:

Statistical Method: The reviewer analyzed the endpoints for cell density, area under the curve, and growth rate 

using mean measured nicosulfuron (active ingredient) concentrations. The reviewer calculated the mean 

measured concentrations as the arithmetic mean of the day 0 and day 4 measured concentrations. The data were 

assessed for normality and homogeneity of variance using Shapiro-Wilk’s and Levene’s tests, respectively. The 

cell density and area under the curve data met all assumptions of normality and homogeneity of variance and 

therefore the NOAEC values based on these endpoints were determined using ANOVA followed by William’s 

tests. The growth rate data met the assumptions of homogeneity of variance, but not normality, and 

transformation of the data did not correct the issue. Therefore, Wilcoxon’s Rank Sum Test was used to 

calculate the NOAEC value based on growth rate. The preceding analyses were all performed using Toxstat 3.5 

statistical software. Despite 100% inhibition at the highest test level, Wilcoxon’s test failed to detect any 

significant differences in growth rate compared to the negative control. Because of the inhibition observed, the 

reviewer deferred to the NOAEC value calculated by the study author in the Executive Summary and 

Conclusions sections of this report. ECx values for cell density and area under the growth curve were 

calculated using probit analysis via Nuthatch statistical software. The ECx value based on growth rate could 

not be determined using the probit method and therefore the reviewer deferred to the study author’s calculated 

EC50 value in the Executive Summary and Conclusions sections

Cell density

EC05: 7.7 mg ai/L 95% C.I.: 5.2 to 11 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 4.31 ± 0.541

Area under the curve

EC05: 6.6 mg ai/L 95% C.I.: 4.6 to 9.4 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 3.75 ± 0.365

Growth rate

EC05: Not calculable 95% C.I.: N/A

EC50: Not calculable 95% C.I.: N/A

NOAEC: 77.95 mg ai/L

Probit Slope: N/A

D. STUDY DEFICIENCIES:

The deviations did not affect the validity of the study. While cyanobacteria may be influenced in changes in 

pH the trends observed in this study did not appear to be pH dependent. Three replicates provided sufficient 

statistical power to result treatment differences and thus arrive at appropriate endpoints and conclusions.

E. REVIEWER’S COMMENTS:

The results of the reviewer generally agree with those of the study author. The reviewer based all analyses on 

mean measured nicosulfuron concentrations, whereas the study author used nominal concentrations only. The 

statistical methods of the reviewer and study author also differed. Therefore, the results of the reviewer and 

study author differ slightly.

The study author did not specify how the NOAEC based on growth rate was determined. It was only noted that 

“appropriate” non-parametric multiple comparison methods were used.
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 15 of 24

The in-life phase of the definitive test was conducted from 16-20 February 2004.

F. CONCLUSIONS: 

This study is scientifically sound and generally meets the guideline requirements for a toxicity test with a 

freshwater cyanobacteria. It is therefore classified as acceptable. The most sensitive endpoints based on mean 

measured concentrations were cell density and area under the growth curve, with NOAEC and EC50 values of 

<4.95 and 18 mg ai/L, respectively.

Cell density

EC05: 7.7 mg ai/L 95% C.I.: 5.2 to 11 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 4.31 ± 0.541

Area under the curve

EC05: 6.6 mg ai/L 95% C.I.: 4.6 to 9.4 mg ai/L

EC50: 18 mg ai/L 95% C.I.: 15 to 22 mg ai/L

NOAEC: <4.95 mg ai/L

Probit Slope: 3.75 ± 0.365

Growth rate

EC05: Not calculable 95% C.I.: N/A

EC50: 39.91mg ai/L* 95% C.I.: 37.68 to 42.14 mg ai/L*

NOAEC: <4.95 mg ai/L**

Probit Slope: N/A

Endpoint(s) Effected: cell density, area under the growth curve, growth rate

*The EC50 and 95% confidence interval based on growth rate were not verified by the reviewer, due to a poor fit of 

the probit model. The values reported here are those calculated by the study author based on nominal nicosulfuron 

active ingredient concentrations.

**This value was based on the study author’s analyses. The study author originally reported the NOAEC based on 

growth rate as <4.68 mg ai/L, based on nominal nicosulfuron exposure concentrations. The reviewer converted the 

NOAEC to the equivalent mean measured nicosulfuron concentration.

III. REFERENCES: 

1. DuPont Haskell Laboratory (2004). Solubility of Nicosulfiiron in Algal Media Solution

(20X AAP) of Nicosulfiiron (DPX-V9360) 75WG plus Dry Surfactant (DPX-GFJ47) (1:6).

Unpublished report, DuPont-14106.

2. Miller, W.E.; Greene, J.C.; Shiroyama, T. The Selenastrum capricornutum Printz Algal

Assay Bottle Test; U.S. Environmental Protection Agency; U.S. Government Printing Office:

Washington, DC, 1978; EPA-600/9-78-018.

3. American Society for Testing and Materials (ASTM). (1990). "Standard Guide for

Conducting Static 96-h Toxicity Tests with Microalgae" in ASTM Annual Book of Standards,

E1218-90. Vol. 11.04, Philadelphia, PA.

4. Environmental Protection Agency, National Eutrophication Research Program (1971). Algal

Assay Procedure: Bottle Test (AAP:BT). National Environmental Research Center,

Corvallis, OR 9330.

5. Snedecor, G.W. and Cochran, W.G. (1967). Statistical Methods. 6th edition. The Iowa State

University Press, Ames.

6. Bruce, R.D. and Versteeg, D.J. (1992). A statistical procedure for modeling continuous
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 16 of 24

toxicity data. Environ. Toxicol. Chem. 11:1485-1494.

7. Hochberg, Y. and. Tamhane, A.C. (1987). Multiple Comparison Procedures, Wiley, New

York.

8. Jonckheere, A.R. (1954). A distribution-free k-sample test against ordered alternatives,

Biometrika41, 133.

9. SAS Software Version 8.2, SAS Institute, Cary, NC, 1999.

10. SAS Software Version 9.1, SAS Institute, Cary, NC, 2004.

11. Slob, W. (2002). Dose-Response Modeling of Continuous Endpoints, Toxicological

Sciences 66, 298-312.

12. Terpstra, T.J. (1952). The Asymptotic Normality and Consistency of Kendall's Test Against

Trend, When Ties are Present in One Ranking, Indag. Math. 14, 327-333.

13. U.S.EPA. (1995). The Use of the Benchmark Dose Approach in Health Risk Assessment,

Risk Assessment Forum, EPA/630/R-94/007, United States Environmental Protection

Agency, Washington, DC. Principal authors: K. Crump, B. Allen, E. Faustman.

14. Westfall, P.H., Tobias R.D., Rom D., Wolfmger R.D. and Hochberg Y. (1999). Multiple

comparisons and multiple tests, SAS Institute, Cary, North Carolina.

APPENDIX I. OUTPUT OF THE REVIEWER’S STATISTICAL VERIFICATION

CELL DENSITY

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY 

 File: 6401C Transform: NO 

TRANSFORMATION

 Shapiro - Wilk's Test for Normality

 ---------------------------------------------------------------------------

-

 D = 694400.0000

 W = 0.9481

 Critical W = 0.8580 (alpha = 0.01 , N = 18)

 W = 0.8970 (alpha = 0.05 , N = 18)

 ---------------------------------------------------------------------------

-

Data PASS normality test (alpha = 0.01). Continue analysis.

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY 

 File: 6401C Transform: NO 

TRANSFORMATION

 Levene's Test for Homogeneity of Variance

 ANOVA Table

 ---------------------------------------------------------------------------

-
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 17 of 24

 SOURCE DF SS MS F

 ---------------------------------------------------------------------------

-

 Between 5 126577.7778 25315.5556 0.7978

 Within (Error) 12 380800.0000 31733.3333

 ---------------------------------------------------------------------------

-

 Total 17 507377.7778

 ---------------------------------------------------------------------------

-

 (p-value = 0.5719)

 Critical F = 5.0643 (alpha = 0.01, df = 5,12)

 = 3.1059 (alpha = 0.05, df = 5,12)

 Since F < Critical F FAIL TO REJECT Ho: All equal (alpha = 0.01)

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY 

 File: 6401C Transform: NO 

TRANSFORMATION

 ANOVA Table

 ---------------------------------------------------------------------------

-

 SOURCE DF SS MS F

 ---------------------------------------------------------------------------

-

 Between 5 111479111.1111 22295822.2222 385.2965

 Within (Error) 12 694400.0000 57866.6667

 ---------------------------------------------------------------------------

-

 Total 17 112173511.1111

 ---------------------------------------------------------------------------

-

 (p-value = 0.0000)

 Critical F = 5.0643 (alpha = 0.01, df = 5,12)

 = 3.1059 (alpha = 0.05, df = 5,12)

 Since F > Critical F REJECT Ho: All equal (alpha = 0.05)

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY 

 File: 6401C Transform: NO 

TRANSFORMATION
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 18 of 24

 William's Test - TABLE 1 OF 2 Ho: Control<Treatment

 ---------------------------------------------------------------------------

-

 ORIGINAL TRANSFORMED ISOTONIZED 

 GROUP IDENTIFICATION N MEAN MEAN MEAN 

 ------ -------------------- --- ----------- ----------- ----------

-

 1 NEG CONTROL 3 6560.0000 6560.0000 6560.0000

 2 4.95 3 5853.3333 5853.3333 5853.3333

 3 9.96 3 3906.6667 3906.6667 3906.6667

 4 20.15 3 3120.0000 3120.0000 3120.0000

 5 40 3 346.6667 346.6667 346.6667

 6 77.95 3 0.0000 0.0000 0.0000

 ---------------------------------------------------------------------------

-

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY 

 File: 6401C Transform: NO 

TRANSFORMATION

 William's Test - TABLE 2 OF 2 Ho: Control<Treatment

 ---------------------------------------------------------------------------

-

 COMPARED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM 

USED

 -------------------- ----------- ----------- ----- ----------- -----------

-

 NEG CONTROL 6560.0000

 4.95 5853.3333 3.5979 * 1.7800 k= 1, 

v=12

 9.96 3906.6667 13.5090 * 1.8700 k= 2, 

v=12

 20.15 3120.0000 17.5142 * 1.9000 k= 3, 

v=12

 40 346.6667 31.6341 * 1.9200 k= 4, 

v=12

 77.95 0.0000 33.3991 * 1.9300 k= 5, 

v=12

 ---------------------------------------------------------------------------

-

 s = 240.5549

------------------------------------------------------------------------

Estimates of EC% 

------------------------------------------------------------------------
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 19 of 24

Parameter Estimate 95% Bounds Std.Err. Lower Bound

 Lower Upper /Estimate

EC5 7.7 5.2 11. 0.079 0.68

EC10 9.3 6.7 13. 0.069 0.71

EC25 13. 9.9 17. 0.053 0.77

EC50 18. 15. 22. 0.037 0.83

 Slope = 4.31 Std.Err. = 0.541 

!!!Poor fit: p < 0.001 based on DF= 3.00 12.0 

------------------------------------------------------------------------

6401C : NICOSULFURON+SURFACTANT A.FLOS-AQUAE CELL DENSITY

------------------------------------------------------------------------

Observed vs. Predicted Treatment Group Means 

------------------------------------------------------------------------

 Dose #Reps. Obs. Pred. Obs. Pred. %Change

 Mean Mean -Pred. %Control

 0.00 3.00 6.56e+03 5.85e+03 713. 100. 0.00 

 4.95 3.00 5.85e+03 5.81e+03 46.1 99.3 0.684 

 9.96 3.00 3.91e+03 5.13e+03 -1.22e+03 87.6 12.4 

 20.1 3.00 3.12e+03 2.55e+03 569. 43.6 56.4 

 40.0 3.00 347. 436. -88.9 7.45 92.6 

 78.0 3.00 0.00 20.8 -20.8 0.356 99.6

AREA UNDER THE CURVE

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE 

 File: 6401A Transform: NO 

TRANSFORMATION

 Shapiro - Wilk's Test for Normality

 ---------------------------------------------------------------------------

-

 D = 151.3920

 W = 0.9909

 Critical W = 0.8580 (alpha = 0.01 , N = 18)

 W = 0.8970 (alpha = 0.05 , N = 18)

 ---------------------------------------------------------------------------

-

 Data PASS normality test (alpha = 0.01). Continue analysis.

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE 
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 20 of 24

 File: 6401A Transform: NO 

TRANSFORMATION

 Levene's Test for Homogeneity of Variance

 ANOVA Table

 ---------------------------------------------------------------------------

-

 SOURCE DF SS MS F

 ---------------------------------------------------------------------------

-

 Between 5 24.3616 4.8723 0.6156

 Within (Error) 12 94.9824 7.9152

 ---------------------------------------------------------------------------

-

 Total 17 119.3440

 ---------------------------------------------------------------------------

-

 (p-value = 0.6906)

 Critical F = 5.0643 (alpha = 0.01, df = 5,12)

 = 3.1059 (alpha = 0.05, df = 5,12)

 Since F < Critical F FAIL TO REJECT Ho: All equal (alpha = 0.01)

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE 

 File: 6401A Transform: NO 

TRANSFORMATION

 ANOVA Table

 ---------------------------------------------------------------------------

-

 SOURCE DF SS MS F

 ---------------------------------------------------------------------------

-

 Between 5 31353.6448 6270.7290 497.0457

 Within (Error) 12 151.3920 12.6160

 ---------------------------------------------------------------------------

-

 Total 17 31505.0368

 ---------------------------------------------------------------------------

-

 (p-value = 0.0000)

 Critical F = 5.0643 (alpha = 0.01, df = 5,12)

 = 3.1059 (alpha = 0.05, df = 5,12)
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 21 of 24

 Since F > Critical F REJECT Ho: All equal (alpha = 0.05)

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE 

 File: 6401A Transform: NO 

TRANSFORMATION

 William's Test - TABLE 1 OF 2 Ho: Control<Treatment

 ---------------------------------------------------------------------------

-

 ORIGINAL TRANSFORMED ISOTONIZED 

 GROUP IDENTIFICATION N MEAN MEAN MEAN 

 ------ -------------------- --- ----------- ----------- ----------

-

 1 NEG CONTROL 3 112.1200 112.1200 112.1200

 2 4.95 3 98.6800 98.6800 98.6800

 3 9.96 3 71.1200 71.1200 71.1200

 4 20.15 3 53.0000 53.0000 53.0000

 5 40 3 9.4800 9.4800 9.4800

 6 77.95 3 0.4000 0.4000 0.4000

 ---------------------------------------------------------------------------

-

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE 

 File: 6401A Transform: NO 

TRANSFORMATION

 William's Test - TABLE 2 OF 2 Ho: Control<Treatment

 ---------------------------------------------------------------------------

-

 COMPARED CALC. SIG TABLE DEGREES OF

 IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM 

USED

 -------------------- ----------- ----------- ----- ----------- -----------

-

 NEG CONTROL 112.1200

 4.95 98.6800 4.6343 * 1.7800 k= 1, 

v=12

 9.96 71.1200 14.1374 * 1.8700 k= 2, 

v=12

 20.15 53.0000 20.3854 * 1.9000 k= 3, 

v=12

 40 9.4800 35.3917 * 1.9200 k= 4, 

v=12

 77.95 0.4000 38.5226 * 1.9300 k= 5, 

v=12

 ---------------------------------------------------------------------------

-
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 22 of 24

 s = 3.5519

------------------------------------------------------------------------

Estimates of EC% 

------------------------------------------------------------------------

Parameter Estimate 95% Bounds Std.Err. Lower Bound

 Lower Upper /Estimate

EC5 6.6 4.6 9.4 0.071 0.70

EC10 8.3 6.1 11. 0.063 0.74

EC25 12. 9.5 15. 0.048 0.79

EC50 18. 15. 22. 0.034 0.85

 Slope = 3.73 Std.Err. = 0.365 

!!!Poor fit: p < 0.001 based on DF= 3.00 12.0 

------------------------------------------------------------------------

6401A : NICOSULFURON+SURFACTANT A.FLOS-AQUAE AREA UNDER CURVE

------------------------------------------------------------------------

Observed vs. Predicted Treatment Group Means 

------------------------------------------------------------------------

 Dose #Reps. Obs. Pred. Obs. Pred. %Change

 Mean Mean -Pred. %Control

 0.00 3.00 112. 102. 9.67 100. 0.00 

 4.95 3.00 98.7 101. -1.98 98.3 1.74 

 9.96 3.00 71.1 85.7 -14.5 83.6 16.4 

 20.1 3.00 53.0 44.6 8.36 43.6 56.4 

 40.0 3.00 9.48 10.4 -0.948 10.2 89.8 

 78.0 3.00 0.400 0.958 -0.558 0.936 99.1

GROWTH RATE

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE GROWTH RATE 

 File: 6401G Transform: NO 

TRANSFORMATION

 Shapiro - Wilk's Test for Normality

 ---------------------------------------------------------------------------

-

 D = 0.0000

 W = 0.7892

 Critical W = 0.8580 (alpha = 0.01 , N = 18)
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 23 of 24

 W = 0.8970 (alpha = 0.05 , N = 18)

 ---------------------------------------------------------------------------

-

 Data FAIL normality test (alpha = 0.01). Try another transformation.

 Warning - The first three homogeneity tests are sensitive to nonnormality

 and should not be performed with this data as is. 

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE GROWTH RATE 

 File: 6401G Transform: NO 

TRANSFORMATION

 Levene's Test for Homogeneity of Variance

 ANOVA Table

 ---------------------------------------------------------------------------

-

 SOURCE DF SS MS F

 ---------------------------------------------------------------------------

-

 Between 5 0.0000 0.0000 2.5136

 Within (Error) 12 0.0000 0.0000

 ---------------------------------------------------------------------------

-

 Total 17 0.0000

 ---------------------------------------------------------------------------

-

 (p-value = 0.0886)

 Critical F = 5.0643 (alpha = 0.01, df = 5,12)

 = 3.1059 (alpha = 0.05, df = 5,12)

 Since F < Critical F FAIL TO REJECT Ho: All equal (alpha = 0.01)

 Title: NICOSULFURON+SURFACTANT A.FLOS-AQUAE GROWTH RATE 

 File: 6401G Transform: NO 

TRANSFORMATION

 Wilcoxon's Rank Sum Test w/ Bonferroni Adjustment Ho: Control<Treatment

 ---------------------------------------------------------------------------

-

 MEAN IN RANK CRIT. SIG

 GROUP IDENTIFICATION ORIGINAL UNITS SUM VALUE REPS 0.05

 ----- -------------------- ----------- ------- ------ ----- ---
Data Evaluation Report on the Acute Toxicity of Nicosulfuron (DPX-V9360) 75WG plus 

Dry Surfactant (DPX-GFJ47) (1:6) to Algae, Anabaena flos-aquae

PMRA Submission Number {........} EPA MRID Number 48376401

Page 24 of 24

 1 NEG CONTROL 0.0676

 2 4.95 0.0664 6.00 None 3 

 3 9.96 0.0622 6.00 None 3 

 4 20.15 0.0598 6.00 None 3 

 5 40 0.0365 6.00 None 3 

 6 77.95 0.0000 6.00 None 3 

 ---------------------------------------------------------------------------

-

 Critical values are 1 tailed ( k = 5 )

 WARNING - There are no critical values for this combination

 of groups and replicates.