Data Evaluation Record on tbe Acute Toxicity ofTebufenozide to Aqu.atic Vascular Plants, 

Lemnagibba 

_ _ _____ _ _ _ _ _ _____________ _::EPA MRJD Number 485330-03 

Data Requirement: 

Test material: Tebufenozide 

Common name 

Chemical name: IUPAC 

EPA DP Barcode 

OECD Data Point 

EPA MRID 

EPA Guideline 

0392144 

OECD Guideline 221 

485330-03 

850.4400 

Purity: 98.6% 

CAS name: 3,5-dimethylbenzoic acid l-(1,1-dimetbylethyl)-2-(4-ethylberzoyl)hydrazide 

CAS No.: 112410-23-8 

Synonyms 

Primary Reviewer: Joan Gaidos Signature: 

Senior Scientist, Cambridge Environmental, Inc. Date: 3/ 15/12 

Primary EPA Reviewer: Andrew Sayer ~ ~~

EP A/OPP/EFEDIERB4 - I -4 Date: 5/10112 

Secondary EPA Reviewer(s): Thomas Steeger, Ph .~ ~~Date: 5/1 011 2 

EPA/OPP/EFEDIERB4 y -~~ 

EPA PC Code 129026 

Date Evaluation Completed: 4/27/ 12 

CITATION: Porch, J. 2011. Tebufenozide: A 7-Day Static-Renewal Toxicity Test with Duckweed (LemnagibbaG3). 

Unpublished study performed by Wildlife International, Ltd., Easton, Maryland. Laboratory project no.: 437 A122A. Study sponsored by Nippon Soda Co., Ltd., Chiyoda-ku, Tokyo, Japan. Study completed July 7, 20 II . 

EXECUTIVE SUMMARY: 

In a 7-day acute toxicity study, the freshwater floating aquatic vascular plants duckweed Le,nna gibba were exposed to 

tebufenozide technical grade active ingredient (98.6% ai) at nominal concentrations ofO (r egative control), 0 (solvent 

[DMF] control), 0.031, 0.063, 0. 13, 0.25, 0.50 and 1.0 mg ai/L under static. renewal conditiors. Measured concentrations 

were <LOQ (<0.0200, negative and solvent [DMF] controls), 0.031, 0.064, 0.13, 0.26, OA5, and 0.94 mg ai/L. All 

comparisons were made relative to the negative control. 

Frond density in the negative control averaged 273 fronds per rep yielding inhibitions of -2, I, I 0, 2, 0 and 9% in the 

mean-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg ai/L treatment groups as compared to the control. The 

NOAEC and EC50 values were 0.94 and >0.94 mg ai/L, respectively (based on mean-meas1tred concentrations). 

Frond biomass (dry weight) averaged 32.4 mg in the negative control, yielding inhibitions of -4, -4, -2, -11, -1 , and 0% in 

the mean-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg ai/L treatment groups, rcspt:ctively. The NOAEC and 

EC50 values were 0.94 and >0.94 mg ailL, respectively (based on mean-measured concentrations). 

Growth rate (based on frond density) averaged 0.446 days"1 in the negative control, yielding inhibitions of -1, 0, 3, I, 0, 

and 3% in the mean-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg ai/L treatment groups, respectively. The 

NOAEC and EC50 values were 0.94 and >0.94 mg a ilL, respectively (based on mean-measttred concentrations). 

After 7 days of exposure, the number of chlorotic fronds observed in the 0.031, 0.064, 0.1 3, 0.26, 0.45 and 0.94 mg/L 

treatment groups was I, I, 0, 0, 2 and 0 representing less than I% of the total number offrords in each group. Necrotic 

Page I of 15 
Data Evaluation Record on the Acute Toxicity of Tebufe.nozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

fronds in the 0.031, 0.064, 0. 13, 0.26, 0.45 and 0.94 mg/L also accounted for less than 1% of the total frond number in 

each group. Plants in the negative and solvent control groups and in all of the treatment groups appeared normal 

throughout the test with no overt signs of toxicity observed. 

This toxicity study is clas ificd as scientifically sound and is consistent with guideline recommendations for a Tier II 

aquatic plant toxicity study using the aquatic vascular plant Lemna gibba. 

Results Synopsis 

Test Organism: Lemna gibba 

Test Type: Static renewal 

Frond density 

ECos: NA 95% C.L: N/A 

EC50: >0.94 mg ai,.L 95% C.l.: N/A 

NOAEC: 0.94 mg ai/L 

LOAEC: >0.94 mg ai/L 

Biomass 

EC05: NA 95% C.L: N/A 

EC50: >0.94 mg aiiL 95% C. I.: NIA 

NOAEC: 0.94 mg aitL 

LOAEC: >0.94 mg ai/L 

Growth rate (based anjroiUI density) 

EC05: NA 95% .: ';:-J/A 

ECso: >0.94 mg ai·'L 95% C.l.: N/A 

NOAEC: 0.94 mg ailL 

LOAEC: >0.94 mg ai/L 

Endpoint(s) Effected: Could not be determined due to a lack of toxicity. 

Most sensitive endpo int: Could not be determined due to a lack of toxicity. 

I. MATERIALS AND METHODS 

GUIDELINE FOLLOWED: This study was conducted according to a protocol based on procedures outlined 

in the U.S. Environmental Protection Agency Series 850 - Ecological Effects 

Test Guidelines (draft), OPPTS Number 850.4400 (1996). Deviations from 

OPPTS 850.4400 were noted: 

COMPLIANCE: 

l. The pH at test initiation was already higher than recommended at 8.0-8.1. By test 

termination the pH was 9.2-9.3; OPPTS guidelines suggest that pH remain in the range 

of7.5 ± 0.1. 

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

These deviations did not impact the acceptability of the study. 

Signed and dated GLP, Data Confidentiality, and Quality Assurance statements 

Page 2 of 15 
Data Evaluation Record on the Acute Toxicity of Tebufenozide to Aqt.atic Vascular Plants, 

Lemnagibba 

-------- -- --------------=EP~A MRIO Number 485330-03 

A. MATERIALS: 

1. Test material 

Description: 

Lot No./Batch No. : 

Purity: 

Stability of compound 

under test conditions: 

Storage conditions of 

test chemicals: 

were provided. This study was conducted in :ompliance with the Good 

Laboratory Practice Standards as published by the U.S. Environmental 

Protection Agency, 40 CFR Part 160 (1989); OECO Principles of GLP 

(ENV/MC/CHEM (98) 17); and Japan MAFF (I I NohSan, Notification No. 

6283, Agricultural Production Bureau, 1999), wilh the following exception: 

Periodic analyses of well water and soil for pote:ttial contaminants were not 

performed in accordance with GLP standards, tut were performed using a 

certified laboratory and standard U.S. EPA analy-ical methods. 

Tebufenozide (TGAI) 

Solid 

6326308 

98.6% 

Method validations using fortified matrix samples yielded recoveries of97. 1 ± 

6.3%. Analytical determinations performed at time 0 yielded recoveries 

ranging from 98.3 to 106% of nominal. At 3 ( oltl solution prior to renewal) 

and 5 (new solution at renewal) days, recoveries were 97.6% to 109% of 

nominal, respectively. At 7 days (old solution at test termination), recoveries 

were 96.2% to II 0% of nominal. Based on these data, the test material was 

stable under the conditions. 

(OECD recommends water solubility, stability in water and light, pKa, Pow, 

and vapor pressure oftest compound) 

The test material was stored at ambient condit10m. in the dark. 

Table 1 lists the chemical physical properties oftebufenozide. 

T a e bl 1 Ph IYSICOC . h em1. 1 ca prope rt' 1es 0 fT e b u ~ enoz• 'd e. 

Parameter Values Comments 

Water solubility 0.83 ppm OP Barcode 

0226011 

Vapor pressure 2.25 X I 0'

8 mm Hg 

OP Barcode 

0226011 

UV absorption Not reported --

pK, Not reported -

4.25 

Page 3 of 15 
Data Evaluation Re, ord on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

Parameter Values Comments 

logKow DP Barcode 

0226011 

2. Test organism: 

Name: Duckweed, Lemna gibba G3 

Strain, if provided: Not reported 

Source: In-house cultures were originally obtained from the U.S. Department of Agriculture and 

maintained in culture medium at Wildlife International, Ltd., Easton, Maryland 

Age of inoculum: The inoculum was taken from a stock culture that had been transferred to 

fresh medium 7 days before testing 

Method of ultivation : Plants were cultured in 20X Algal Assay Procedure (AAP) culture medium 

B. STUDY DESIGN: 

I. Experimental Conditions 

a. Range-finding study: None reported. 

b. Definitive Study 

T a e bl 2 : E xper.menta I P · arameters 

Parameter Details Remarks 

-------------------- Criteria 

- Acclimation period: 7 days 

Culturing media and conditions: Duckweed were acclimated to the 

(same as test or not) culture medium; the culturing 

conditions were the same as test 

Health: (any mortality observed) Not reported 

Test system Fronds were transferred to newly 

Static/static renewal Static-renewal prepared test solutions on days 3 and 

5. 

Renewal rate for static rt·mewal 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). 

Test vessels were placed in a 

Incubation facility temperature-contr?lled environmental 

chamber. 

Page 4 of 15 
Data Evaluation Record on the Acute Toxicity of Tebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

Parameter 

Duration of the test 

Test vessel 

Material: (glass/stainless steel) 

Size: 

Fill volume: 

Details of growth medium name 

pH at test initiation: 

pH at test termination: 

Chelator used: 

Carbon source: 

7 days 

Glass 

250mL 

100 rnL 

8.0-8. I 

9.2-9.3 

Yes 

NaHC03 

Details 

If non-standard nutrient medium N/A 

was used, detailed composition 

provided (Yes/No) 

Dilution water 

source/type: 

pH: 

water pretreatment (if any): 

Total Organic Carbon: 

particulate matter: 

metals: 

pesticides: 

chlorine: 

Purified well water 

Adjusted to 7.5 ± 0. I if necessary 

Water was sterilized 

Not reported 

Not reported 

Not reported 

Not reported 

Not reported 

Page 5 of 15 

EPA MRJD Number 485330-03 

Remarks 

Criteria 

EPA requ.res a duration of 14 days. 

Seven day studies will be accepted for 

review by tl,e Agency. 

EPA recommends the following 

culture media: Modified Hoagland's 

E+ or 20X-AAP. Chelating agents (e.g. 

EDTA) are recommended in the 

nutrient medium for optimum cell 

growth. l ower concentrations of 

chelating agents (down to one-third 

of the normal concentration 

recommended for AAP medium) may 

be used ir. the nutrient medium used 

for test SOJ'ution preparation if it is 

suspected that the chelator will 

interact w 'th the test material. ASTM 

reference, £1415-9/and D 3978-80 

(reapproved 1987). 

EPA rt?COt1mends a pH of - 5.0. A 

solution p!J of 7.5 is acceptable if type 

20X-AAP n ltrient media is used. 
Data Evaluation Record on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRJD Number 485330-03 

- Parameter Details Remarks 

-------------------- Criteria 

Indicate how the test material is A primary stock solution was Concentration of DMF in solvent 

added to the medium (added prepared by mixing a calculated control was 0.1 mL DMF/L. 

directly or used stock so ution) amount (not reported) of test material 

into solvent dimethylformamide 

(DMF) and bringing it to volume with 

20X AAP medium in a 500 mL 

vessel. 

Aeration or agitation Neither 

Sediment used (for root2d aguatic NIA 

vascular plants) 

Origin: 

Textural classification (~'Osand, silt, 

and clay): 

Organic carbon(%): 

Geographic location: 

Number of reQiicates 

Control: 3 

Solvent control: 3 

Treatments: 3 

Number of plants/replicRte 12 plants per replicate 

-------------------- EPA requires 5 plants. 

Number of fronds/plant 4 fronds per plant 

-------------------- EPA requires 3 fronds per plant. 

Test concentrations 

Nominal: 0 (negative control), 0 (solvent -------------------- control), 0.031, 0.063, 0.13, 0.25, EPA requires at least 5 test 

0.50, and 1.0 mg ai!L concentrations with a dose range of2X or 

Mean-Measured: 3X progression. 

<LOQ ( <0.0200, negative and solvent 

[DMF] control), 0.031, 0.064, 0. 13, 

0.26, 0.45, and 0.94 mg ai/L 

Solvent (type, percentage, if used) Dimethylformamide (DMF), 0.1 mL 

DMF/L 

Page 6 of 15 
Data Evaluation Record on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRJD Number 485330-03 

Parameter Details Remarks 

-------------------- Criteria 

- Method and interval of analytical Samples taken from day 0, day 3 (old) 

verification and day 5 (spent) solutions and day 7 

spent solutions were analyzed via 

HPLC with UV detection (wavelength 

of220 run). 

Test conditions 

Temperature: 24.0-26.7°C 

Photoperiod: Continuous 

Light intensity and quality: 6220-6790 lux 

Warm-white fluorescent bulbs 

Reference chemical (if used} 

name: N/A 

concentrations: 

Other parameters, if any None 

2. Observations: 

T ba l e 3 : Ob servahon parameters 

I Parameters I Details I Renarks/Criteria I 

Parameters measured (e.g.,: - frond density 

number of fronds, plant dry weight -biomass (dry weight) or other toxicity symptoms) -growth rate (frond density) 

- growth rate 

- frond condition 

Measurement technique for frond Frond density was determined via 

number and other end points direct counting. Frond condition 

was visually determined. 

Biomass was measured by drying 

the plants at 60°C for ca. 48 hours 

prior to weighing. Growth rates 

were determined using equations 

that took into account frond 

number or biomass at the 

beginning and end of the test. 

Observation intervals Frond density and condition were 

determined at days 0, 3, 5, and 7. 

Page 7 of 15 
Data Evaluation Re ord on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

I Parameters 

= 

I Details I Remarks/Criteria I 

Biomass was determined at study 

initiation and termination. 

Other observations, if any None 

Indicate whether there ""'as an Yes; frond density was 273 fronds 

exponential growth in the control per replicate in the control at 7 

days. 

Were raw data included? Yes 

II. RESULTS and DISCUSSION: 

A. INHIBITORY EFFECTS: 

According to the study authors, frond density in the negative control averaged 273 fronds per rep yielding inhibitions 

of -2, I, I 0, 2, 0 and 9% in the mean-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg ai/L treatment groups as 

compared to the control (Table 4). Percent inhibition was based on pooled controls. The study author-reported 

NOAEC and ECso va' ues were 0.94 and >0.94 mg ai/L, respectively (based on mean-measured concentrations). 

Frond biomass (dry weight) averaged 32.4 mg in the negative control, yielding inhibitions of -4, -4, -2, -II, -I , and 

0% in the mean-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg ai/L treatment groups, respectively (Table 

4a). The study author-reported NOAEC and EC50 values were 0.94 and >0.94 mg ai/L, respectively (based on 

mean-measured conetmtrations). 

Growth rate (based on frond density) averaged 0.446 days·• in the negative control, yielding inhibitions of -1, 0, 3, I, 

0, and 3% in the mea:1-measured 0.031, 0.064, 0.13, 0.26, 0.45, and 0.94 mg a ilL treatment groups, respectively 

(Table 4a). The stud)- author-reported NOAEC and EC50 values were 0.94 and >0.94 mg a ilL, respectively (based on 

mean-measured concentrations). 

After 7 days of expo5-ure, the number of chlorotic fronds observed in the 0.031, 0.064, 0.13, 0.26, 0.45 and 0.94 

mg!L treatment groups was 1, I, 0, 0, 2 and 0 representing less than I %of the total number of fronds in each group. 

Necrotic fronds in the 0.031 , 0.064, 0.13, 0.26, 0.45 and 0.94 mg!L also accounted for Jess than I% of the total 

frond number in each group. Plants in the negative and solvent control groups and in all of the treatment groups 

appeared normal throughout the test with no overt signs of toxicity observed. 

Page 8 of 15 
Data Evaluation Record on the Acute Toxicity of Tebufenozide to AqtJ.atic Vascular Plants, 

Lemna gibba 

EPA MRID Number 485330-03 

T a bl e 4 Effi f b fi f d : ect o te u enozt e on ron num b er o fd uc k d wee ., L emna g• ibb a * 

Treatment Mean-measured (and Frond number/replicat e (mean) 

Initial frond 

nominal) concentration number/test 7 days 

mg ai/L solution 3 days 5 days 

frond m·mber % inhibition• 

Negative control 12 35 95 273 N/A 

Solvent control 12 34 90 260 N/A 

0.031 (0.031) 12 39 104 279 -2 

0.064 (0.063) 12 37 93 270 l 

0.13 (0.13) 12 36 93 247 10 

0.26 (0.25) 12 35 90 269 2 

0.45 (0.50) 12 34 92 27t 0 

0.94 ( 1.0) 12 34 88 24£ 9 

.. . * Percent mhibitlon based on pooled controls 

a Percent inhibition based on comparison to negative control. 

Table 4a: Effect of tebufenozide on growth rate and dry weight of duckweed, L emna gibba * 

Growth rate Treatment Mean-measured Growth rate Dry weight Initial frond based on frond D ·y weight % % (and nominal) concentration mg number/test density Inhibition • (mg, mean) a Inhibition • ilL solution (days ·

1

, mean) 

Negative control 12 0.446 N/A 32.4 NIA 

Solvent control 12 0.439 N/A 32.2 N/A 

0.031 (0.031) 12 0.449 -1 33.9 -4 

0.064 (0.063) 12 0.445 0 33.6 -4 

0.13 (0.13) 12 0.431 3 33.1 -2 

0.26 (0.25) 12 0.444 1 35.9 -11 

0.45 (0.50) 12 0.446 0 32.8 -1 

0.94 ( 1.0) 12 0.432 3 32.4 0 

. . . * Percent mhibition based on pooled controls 

a Percent inhibition based on comparison to negative control. 

Page 9 of 15 
Data Evaluation Re ord on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

Table 5: Statistical endpoint values (study author-reported) 

Statistical Endpoint Frond Density Biomass Growth rate (based on frond 

density) 

NOAEC (mg ai/L) 0.94 0.94 0.94 

ECos (95% CI; mg ai/L) N/A N/A N/A 

LOAEC (mg ai/L) >0.94 >0.94 >0.94 

EC50 (95% CI; mg ai/L) >0.94 >0.94 >0.94 

B. REPORTED STATISTICS: 

The study authors analyzed frond density, biomass, and growth rate (based on frond density and biomass) using 

mean-measured concentrations and SAS® (SAS Institute, Cary, NC; Version 8.2). The data were tested for 

nonnality and homog.::neity of variance (p $ 0.05) using Shapiro-Wilks' and Levene's tests, respectively. The data 

were then analyzed u5-ing ANOV A and Dunnett's t-test. Since there was less than 50% inhibition in endpoints in any 

treatment group, day 7 EC, values were not calculated. Inhibitions were determined based on the negative controls. 

Page 10 of 15 
Data Evaluation Record on the Acute Toxicity ofTebufenozide to Aqu:atic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

C. VERIFICATION OF STATISTICAL RESULTS: 

Statistical Method: The reviewer analyzed frond density, biomass, and growth rate data using both initial-measured 

and mean-measured concentrations for analysis. The replicate data were frrst tested for normality using the Shapiro 

Wilks' tests and for homogeneity of variances using Levene's test in Toxstat 3.5. All endpoint data met the 

assumptions of ANOV A, and were thus analyzed using Dunnett's test to determine the NOAEC. There were no 

statistically significant inhibitions (a=0.05) compared to the negative control for any <mdpoint. Analysis of frond 

density at 7 days yielded no significant differences (a=0.05). The EC50 values for each of the endpoints were 

empirically determined to be greater than the highest concentration tested. The reviewer's results, based on the 

statistical verification presented in Appendix I, are presented in the Executive Summary and Conclusions sections of 

this data evaluation record (DER). 

Based on Mean-measured Concentrations: 

Frond density 

ECos: NA 95% C.l.: N/A 

ECso: >0.94 mg ai/L 95% C. I.: N/A 

NOAEC: 0.94 mg ai/L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Biomass 

ECo5: NA 95% C.I.: N/A 

ECso: >0.94 mg ai/L 95% C. I.: N/A 

NOAEC: 0.94 mg ai/L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Growth rate (based on f rond density) 

EC05: NA 95% C.l.: N/A 

EC5o: >0.94 mg ai/L 95% C.I.: N/A 

NOAEC: 0.94 mg ai/L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Endpoint(s) Effected: Could not be determined due to a lack of toxicity. 

Most sensitive endpoint: Could not be determined due to a lack of toxicity. 

D. STUDY DEFICIENCIES: 

None. 

E. REVIEWER'S COMMENTS: 

The reviewer's and the study authors' results were in agreement. The most sensitive endpoint could not be 

determined due to lack of toxicity. The reviewer's results (and the study author's ECsJ value for growth rate) are 

presented in the Executive Summary and Conclusions sections of this data evaluation 'ecord (DER). 

The in-life phase of the study was conducted from May 20 to May 27, 20 II. 

F. CONCLUSIONS: 

Page II of 15 
Data Evaluation Reeord on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagihha 

EPA MRID Number 485330-03 

This study is scientif:cally sound and classified as ACCEPT ABLE. The most sensitive endpoint could not be 

determined due to a lack of toxicity. The 7-day NOAEC and EC50 values were 0.96 and >0.96 mg ai/L, based on 

mean-measured concentrations and the absence of statistically significant effects at the highest concentration tested .. 

Based on Mean-measured Concentrations: 

Frond density 

EC05: NA 95% C.L: N/A 

EC50: >0.94 mg aiiL 95% C.I.: N/A 

NOAEC: 0.94 mg ai1L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Biomass 

ECo5: NA 95% C.l.: N/A 

EC50: >0.94 mg ai1

L 95% C.l.: N/A 

NOAEC: 0.94 mg ai'L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Growtlr rate (based on f rond density) 

EC05: NA 95% C. I.: N/A 

EC50: >0.94 mg ai/L 95% C.l.: NIA 

NOAEC: 0.94 mg ai/L 

LOAEC: >0.94 mg ai/L 

Probit Slope: Could not be determined 

Endpoint(s) Effected: Could not be determined due to a lack of toxicity. 

Most sensitive endpoint: Could not be determined due to a lack of toxicity. 

III. REFERENCES: 

Organization for Economic Cooperation and Development. 2006. Guidelines for Testing of Chemicals, Guideline 

221: Lemna sp., Growth Inhibition Test. Adopted 23 March 2006. 

U.S. Environmental Protection Agency. 1996. Series 850 - Ecological Effects Test Guidelines (draft), OPPTS 

Number 850.4400: Aquatic Plant Toxicity Test using Lemna spp., Tiers I and II. Washington, D.C. 

ASTM Standard Guicle E 1415-91 E. 1991. Standard Guide for Conducting Acute Toxicity Tests with Lemna gibba 

GJ. American Society for Testing and Materials, Philadelphia, PA. 

Microsoft Corporation, Microsoft Excel2000. Copyright 1985- 1999. 

West, Inc. and David D. Gulley, 1996. TOXSTAT Version 3.5. Western Ecosystems Technology, Inc. Cheyenne, 

Wyoming. 

Page 12 of 15 
Data Evaluation Record on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRJD Number 485330-03 

APPENDIX I. OUTPUT OF REVIEWER'S STATISTICAL VERIFICATION: 

Title: 3303 frond no . 7 day 

File: 3303fd Transform: 

Shapi ro - Wi l k' s Test for Normality 

D 8587.3333 

w = 0.9363 

Critical w 

w 

0 .8840 (alpha 

0 .9160 (alpha 

0.01 

0 .05 

N 

N 

24) 

24 ) 

NO TR\NSFORMATION 

Data PASS norma lity t est (alpha = 0.01). Continue analysi s. 

Title : 3303 frond no . 7 day 

File: 3303fd Transform: NO TRJ;NSFORMATION 

Levene' s Test for Homogeneity of Variance 

SOURCE 

Between 

Withi n (Error ) 

Total 

Critical F 

ANOVA Table 

OF ss 

7 535. 3333 

16 6124 .6667 

23 6660.0000 

4 . 025 9 (alpha = 0 . 01 , df 

2.6572 (al pha = 0 .05, df 

7, 16) 

7, 16) 

MS F 

76.4762 0.1998 

382.7917 

(p-value = 0.9809) 

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

Title: 3303 frond no. 7 day 

File: 3303fd Transform : NO TRANSFORMATION 

ANOVA Table 

---------------------------------------------------------------·-------------

SOURCE OF ss MS F 

------- --- -- ----- --- ------ --- ---- -- -- -- -- ----- -- ---- -- --- --·~- ---- ---- --- Between 7 3071.6250 4 38.8036 0.8176 

With in (Error ) 16 8587. 3333 536.7083 

--------------------------------------------------------------··------------- Tota l 23 11658. 9583 

--------------------------------------------------------------··------------- (p - val ue = 0 . 5864) 

Page 13 of 15 
Data Evaluation Re•;ord on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

Critical F 4 . 0259 (alpha = 0 .01, df = 7,16) 

2 . 6572 (alpha = 0.05, df = 7, 16) 

Since F < Cri t ical F FAIL TO REJECT Ho: All equal (alpha 

Titl e : 3303 fron1 no . 7 day 

0 . 05) 

Fil e : 3303fd Transform: NO TRANSFORMATION 

Dunnett's Te st TABLE 1 OF 2 Ho :Control<Treatment 

TRANSFORMED MEAN CALCULATED IN SIG 

GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT 0 . 05 

-------------------- ----------- ------------------ 1 Neg. Control 273 . 3333 273 . 3333 

2 solven:: control 259 . 6667 259 . 6667 0.7225 

3 0 . 031 2 79 . 0000 279.0000 -0.2996 

4 0 . 064 2 70 . 3333 270.3333 0.1586 

5 0 .13 247 . 0000 247.0000 1 . 3921 

6 0 . 26 268.6667 268 . 6667 0.2467 

7 0 . 45 273 . 6667 273.6667 -0 .0176 

8 0 . 94 248.0000 248.0000 1 .3393 

Dunnett critical value = 2 . 5600 (1 Ta iled, alpha = 0.05, df = 7,16) 

Titl e: 3303 frond no . 7 day 

File: 3303fd Transform: NO TRANSFORMATION 

Dunnett's Test. TABLE 2 OF 2 Ho:Control <Treatment 

-------------------·-------------------------------------------------------- NUM OF MIN SIG DIFF 

GROUP IDENTIFICATION REPS (IN ORIG . UNITS) 

-------------------- ------- ------- --------- 1 Neg. Control 3 

2 sol ven:: control 3 48 .4243 

3 0.031 3 48 .4243 

4 0.064 3 48 . 4243 

5 0. 13 3 48.4243 

6 0 . 26 3 48.4243 

7 0 . 45 3 48.4243 

8 0 . 94 3 48 . 4243 

TEBUFENO.TXT : 3303 frond no. 7 day (Vi a Nuthatch) 

Wi lliams Test 

[One -Sided Test for Decrease, alpha = 0 .050000 l 

Dose 

0 

0.031 

0 . 06 4 

0 . 13 

0 .26 

0 . 45 

Isotone 

Means 

271 

271 

27::> 

263 

263 

263 

T- bar P- value Significance 

- 0.258 

- 0.2374 

0.2098 

0.2098 

0 . 2098 

N.S. 

N.S. 

N.S . 

N.S. 

N.S. 

Page 14 of 15 

% OF DIFFERENCE 

CONTROL FROM CONTROL 

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

17 . 7 13.6667 

17 . 7 -5. 6667 

17 . 7 3 . 0000 

17 . 7 26 . 3333 

17. 7 4 . 6667 

17 .7 - 0 . 3333 

17. 7 25 . 3333 
Data Evaluation Record on the Acute Toxicity ofTebufenozide to Aquatic Vascular Plants, 

Lemnagibba 

EPA MRID Number 485330-03 

0.94 248 1. 146 N.S. 

"* " =Signifi cant; "N . S."=Not Significant . 

Page 15 of 15