### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 Data Requirement: 

### PMRA Data Code EPA DP Barcode OECD Data Point EPAMRID EPA Guideline { 

### } 

### 361296,358877,358880,358882,358885 { 

### } 

### 47643006 850.1300 95.8-96.8% Purity: 

### IN L5300-20 Tribenuron-methyl IUPAC: methyl 2-[4-methoxy-6-methyl-l,3,5-triazin-2-yl(methyl) carbamoylsulfamoyl]benzoate CAS name: methyl 2-[[[[(4-methoxy-6-methyl-l,3,5-triazin-2-yl) methylamino]carbonyl]amino]-sulfonyl]benzoate CAS No.: 101200-48-0 Synonyms: DPX-L5300-20; Haskell No. 15,527 Test material: 

### Common name: Chemical name:

Primary Reviewer: 

### Christie E. Padova StaffScientist, Dynamac Corporation Secondary Reviewer: 

### Teri S. Myers Senior Scientist, Cambridge Environmental Inc. Reviewer: 

### ChristinaWendel USEPA/OCSPP/OPP/ERBII/Biologist Signature:

# {!~ 

# t. 

# P J..nr-- Date: 

### 06/16/2011 Signature:

## ~'5 Date: 

### 07/12/2011. Signature: 

# W Date: 

### 12/06/2011 Reference/Submission No.: {. } Company Code Active Code Use Site Category: 

### EPA PC Code 

### { } [For PMRA] { } [For PMRA] { } [For PMRA] 128887 Date Evaluation Completed: 

### 12/06/2011 CITATION: 

### Hutton, D.G. 1989. Chronic Toxicity of IN L5300-20 to 

### Daphnia magna. Unpublished study performed by E.!. du Pont de Nemours and Company, Inc., Newark, DE, and sponsored by E.!. du Pont de Nemours and Company, Wilmington, DE. Laboratory Study No. 164-89. Study initiated February 22,1989 and completed April 10, 1989.

Page 1 of29 

# '1111.1111 2086464

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 EXECUTIVE SUMMARY: The 21-day-chronic toxicity of IN L5300-20 (Tribenuron-methyl) to 

### Daphnia magna was studied under static-renewal conditions. Daphnids were exposed to IN L5300-20 at nominal concentrations of

# °(negative control), 31, 62, 125, 250, 500, and 1000 mg/L. Mean-measured concentrations were <17.5 «LOQ, control), 32, 56, 120,250,480, and 940 mg ai/L. The 21-day ECso for immobilization was >940 mg ai/L, the highest concentration tested. The reviewer concluded that a NOAEC could not be determined in this study due to suggestive treatmentrelated effects at the lowest treatment level (32 mg ai/L). Significant reductions (p<0.05) were detected for total offspring production at the 32, 120, 250, 480, and 940 mg ai/L treatment levels and for adult length at the 32, 250, 480, and 940 mg ai/L levels; while the responses were not entirely linear, both endpoints were significantly reduced at the lowest level suggesting that the effect may have been treatment-related. A treatment-related delay of first brood release was additionally indicated at the 940 mg ai/L level and the number ofoffspring produced/day per surviving adult was adversely affected at the 250, 480, and 940 mg ai/L treatment levels. The 21-day ECsobased on reproduction parameters was 380 (330 -440) mg ai/L for total offspring and 410 (350 - 470) mg ai/L for offspring produced/dayper surviving adult. Following 21 days ofexposure, survival averaged 85, 95, 80, 85, 78, 82, and 70% in the control and mean-measured 32, 56, 120, 250, 480, and 940 mg ai/L treatment levels, respectively. The study author's analysis detected a significant difference (p<0.05) compared to the negative control at the 940 mg ai/L level, while the reviewer's analysis did not detect any differences for this endpoint. The mean time to first brood release was 9.3 days for the negative control, and 10.4, 10.1, 10.0, 9.7, 8.8, and 15.6 days for the mean-measured 32, 56, 120,250,480, and 940 mg ai/L treatment levels, respectively. The mean number ofoffspring per surviving parent averaged 116, 85, 115, 98, 89, 36, and 2 for the control and mean-measured 32, 56, 120, 250, 480, and 940 mg ai/L levels, respectively. Mean lengths were 3.7, 3.6, 3.6, 3.6, 3.6, 3.5, and 3.3 mm for the control, and mean-measured 32, 56, 120, 250, 480, and 940 mg ai/L levels, respectively. No males, ephippia, or immobilized young were observed in the control group or any treatment level during the study, and no eggs were observed in the bottom ofany test vessel. This study is scientifically unsound

### and does not 

### satisfy the guideline requirement for a chronic toxicity study with freshwater invertebrates. The dissolved oxygen was <60% (equivalentto 5.5 mg/L at 20°C; reviewer-calculated) for anunspecifiedperiod oftime at all levels during the study. In addition, the resulting analysis ofthe data indicated that a NOAEC could not be determined, as treatment-related effects were seen in the lowest test concentration. Therefore, based on these analyses, the results from this study are not reliable, and are not acceptable for use within ecological risk assessments. This study is classified as 

### invalid.

Page 2 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 

### Results Synopsis Test Organism Age (eg. l " instar): First instar, <24 hours old Test Type (Flow-through, Static, Static Renewal): Static-renewal 21-day ECso

### (mortality/immobility): >940 mg ai/L 21-day ECso

### (reproduction): 380 mg ai/L Slope: 5.07±0.724 NOAEC: <32 mg ai/L LOAEC: 32 mg ai/L 95% C.L: N/A 95% C.L: 330-440 mg ai/L

Endpoints affected: Survival, time to first brood release, offspring production, and growth (length) Most sensitive endpoint(s): Total offspring production and growth (length) I. 

### MATERIALS AND METHODS GUIDELINE FOLLOWED: This study was conducted following guidelines outlined in OECD Guideline No. 202 (1984) and U.S. EPA Standard Evaluation Procedure, Daphnia magna 

### Life-Cycle (21-Day Renewal) Chronic Toxicity Test" (1986). Deviations from OPPTS 850.1300 (draft, 1996) guidance included: I. A NOAEC could not be determined in this study due to suggestive adverse effects on total offspring production and adult length at the lowest treatmentlevel. 2. Dissolved oxygen in the test vessels ofaged solutions was <60% (equivalentto 5.5 mg/L at 20°C; reviewer-calculated) for an unspecifiedperiod oftime at all levels during the study. 3. The source ofthe dilution water used in the toxicity test was filtered well water that had been previously used for fathead minnow culturing in a flow-through system within the test laboratory, and filtered through a (l.Sum 

### filter prior to use in the daphnid toxicity study. The water was not tested for the presence ofammonia, therefore it is not known if that may have had a negative impact on the health ofthe test organisms. 4. The type and quantity offood offered at each renewal period was not reported. 5. Although a I N NaOH solution was used to enhance solubility ofthe test substance, a solvent control group was not included in the study. Furthermore, it was unknown if concentrations ofNaOH were the same for all levels. These deviations do 

### affect the acceptability ofthe study.
Page 3 of29

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates

-

### Daphnia sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 COMPLIANCE: Signed and dated GLP (1989), Quality Assurance (1989), and Data Confidentiality claims (2008) statements were provided. A. MATERIALS: 

### 1. Test Material IN L5300-20 (Tribenuron-methyl) Description: Notreported Lot No./Batch No. : Not reported Purity: 95.8 - 96.8% Stability ofcompound under test conditions: The stability oftribenuron-methyl was verified analytically at all levels weekly throughoutthe study. For all levels, replicate results were within ±20%. Storage conditions of test chemicals: Not reported th I fT 'b ti I IYSICOC 

### emica proper res 0 rr 

### enuron-me ryr, 

### Parameter Values Comments Water solubility at 20°C Not reported Vaporpressure Not reported UVabsorption Not reported pKa Not reported Kow Not reported Ph . h



### (GECD recommends watersolubility, stability in waterandlight, pKa, Pow, andvapor pressure of test compound) 2. Test Organism: Species: 

### Daphnia magna, first instar, <24 hours old 

### EPA andGECD recommendDaphnia magna Age ofthe parental stock: 25 days old 

### EPA recommends thatyoungdaphnids**~24** 

### hours old from a separateparental culture be used Page 4 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -

### Daphnia sp. 

### PMRA SubmissionNumber { } EPA MRID No. 47643006 Source:

B. STUDY DESIGN: 

### Haskell Laboratorybred culture EPA requires all test organisms mustbeproduced from laboratory rearedculture that has been maintainedforat least 

### 21 days at test conditions in dilution water with renewal of the culture medium at least three timesperweek.

1. Experimental Conditions



### a. Range-finding Study: Nominal test levels for the definitive study were based upon acute toxicity data determined prior to study initiation. The acute, static ECso (immobilization) was 720 

### mg/L for unfed Daphnia magna 

### and>1000 

### mg/L for fed Daphnia magna. 

### b. Definitive Study Table 1: Experimental Parameters Remarks Parameter

Parental acclimation: Period: Conditions: (same as test or not) Feeding: Health (any mortality observed): 

### Details

25 days Same as test Troutchow (Glencoe) and yeast (Fleischmann's) No sickness, injuries, or abnormalities were noted in the test organisms prior to initiatingthe test.



### Page 5 of29 Criteria Daphniawere cultured in 2-L beakers containing ca. 1.8 L of waterand 25 daphniaper beaker. EPA recommends thatpriorto testing, daphnids that are at least 10-12 days old(those that have hadat leastone brood) shouldbe separated from the culture, put in separate containerand maintained/orat least 

### 21 days to insure thatgoodhealth conditions arepresent

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


### Remarks Parameter Details Criteria Test condition: static renewal/flow-through: Type ofdilution system- for flow through method.

Renewal rate for static renewal

Aeration, ifany

Duration ofthe test Static-renewal

N/A Three times per week

No supplemental aeration was provided.

21 days The parent Daphnia were transferred into fresh test solutions containing food three times per week. (EPA requires consistent.flow rate of 5-10 vol124hours, meter systems calibratedbefore study andcheckedtwice daily during testperiod)

EPA recommends test chambers shouldnot be aerated

Recommendedduration is 

### 21 days. Test vessel Material: (glass/stainless steel) Size (for growth and reproduction/survival test): Fill volume: Glass

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

### 1. RecommendedMaterial: Glass, No. 

### 316 stainless steel, or 250 mL (6.1-cm diameter) per.fluorocarbonplastics 2. RecommendedSize: 250 ml Ca. 200 mL (6.8-cm depth) with 200 mlfillvolume;100ml with 80 mlfill volume DECD guideline recommends thatparent animals be maintained individually; onepervessel, with 50

- 100 ml of medium in each vessel.

Page 60f29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


### Remarks Parameter Details

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

### Criteria Source ofdilution water The dilution water used for Results from periodic analysis of testing was filtered fish tank the well water were provided; water (laboratory well water waterquality measurements ofthe thathad been used for dilution water were not taken after fathead minnow culturing in use in the fish culturing. a flow-through system, then filtered through a 0.8-f.lm The use ofold fish culture water filter). Water was originally as the dilution water is a notable obtained from a 350-foot deviation. It is unknown ifthe encased (in bedrock), and water contained ammonia as aerated and filtered (IO urn) measurements were not taken. priorto use in fish culturing. Therefore it is not known if ammonia was present in the dilution waterused in the Daphnia toxicity test; or ifit may have had a negative impacton the health ofthe test organisms.

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

Recommendedsource of dilution water includes unpollutedwell or springwaterthat has been tested for contaminants, or appropriate reconstitutedwater (seeASTM for details). Water parameters: The pH offresh solutions Hardness 75 ± 2 mg/L as CaC03 increased with increasing test concentrations. pH Fresh soln.: 7.2 to 8.8 Aged soln.: 7.1 to 8.2 DO was <60% (equivalentto 5.5 mg/L at 20°C; reviewerDissolved oxygen Fresh soln.: 8.2 to 8.7 mglL calculated) for an unspecified Aged soln.: 4.6 to 7.4 mglL period oftime at all levels during the study. Temperature 19.7 to 20.4°C Alkalinity was 82 ± 2 mglL as Total Organic Carbon Not reported CaC03, and conductivity was 169 Particulate matter Not reported ± 2 umbos/em. Inorganic ana1ytes Not reported Pesticides Not reported Raw water chemistry datawere Chlorine Not reported not provided. However, the chemical characteristics ofthe

- - - - - -------- - - - ----------- - --------- - - -- Page 

### 7 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


# I Parameter 

# I Details 

# I

... n .n ••

# R;:::;

.......

n. n 

# ··1

Frequency ofmeasurements Dissolved oxygen (DO) was Haskell Laboratory Well Water measured at each renewal are as follows: TOC: <1.5 mg/L; period in one old and one COD: <17mg/L; Solids, total new test solution from all suspended (TSS): <4 mg/L; levels. The pH was Pesticides: None detected (but measured at each renewal detection level was not reported); period in one old and one Chloride: 3.7 mg/L new test solution from the ----- - - - - - - - - - - - - - - - - - - - - ---- - - - - - - - - - - - control, 31 (low), 500 Recommendedhardness: 160 to (medium), and 1000 (high) 180 mg/L as CaC03; OECD treatment levels. The recommends>140 mg/L as temperature was CaC03 continuously monitored (not Recommended pH: 

### 7.6 to 8.0pH further specified) and shouldnot deviate by more than manually checked each 1.

# aunit for more than 

### 48 hours. workday. The hardness, OECD recommends thatpHrange total alkalinity, and be 6

- 9 anddoes not vary more conductivity ofthe control than 

### 1.5 units in anyone test. dilution water were Recommendeddissolvedoxygen: measured once weekly renewalshouldnot drop below during the test. 50%formore than 

### 48 hours. Recommended flow-through: ?,60%throughout test. Recommendedtemperature: 20 DC ±

# rc; shouldnot deviate from 20 DC by more than 

### 5 DC for more than 

### 48 hours. OECD recommends a range of

### 18 - 22°C; temperature shouldnot vary more than ± 2°C OECD guideline recommends that total organic carbon <2 mg/L Number ofreplicates: There were six toxicant levels For growth and reproduction: 10replicates per level spaced by a separation factor of2.

------------------------------------------- For survival test: (same) Number of replicates should include a control(s) andat least 5 test concentrations; dilution factor shouldnot be greater than 50%. OECD recommends that at least 5 Page 

### 8 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


### Remarks Parameter Details ------------------------------------------- 

### Criteria
test concentrations be used in a geometric series with a separation factor not exceeding 3.2. Number oforganisms: Study design follows that For growth and reproduction: Four organisms per replicate described in OPPTS guidance for a flow-through study. All For survival test: (same) replicates were observed for survival, reproduction, and growth.

- - - - - ------ - - - - - ------ - - - - - ------ - - ------- Recommendednumberof organisms include 22 daphnids/test concentration; 7 test chambers shouldcontain 1 daphnideach, and

### 3 test chambers contain 

### 5 daphnids each. OEeD recommends holding a minimum of 10 daphnids individually forstatic tests. Forflow-through tests, 40 animals shouldbe dividedinto 4 groups of 

### 10animals at each test concentration.



### Page 9 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia sp. 

### PMRA SubmissionNumber { } EPA MRID No. 47643006
Remarks Parameter Details

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

### Criteria Treatment Concentrations: It was not reported ifthe nominal nominal: 

# ° (negative control), 31, 62, concentrations were corrected for 125,250,500, and 1000 purity ofthe test substance. mg/L Concentrations ofTribenuronmean-measured: <17.5 «LOQ, control), 32, methyl were determined in fresh 56, 120,250,480, and 940 solutions (all levels) on Days 0, 7, mg ai/L and 14 and in aged ('old') solutions (all levels) on Days 7, 14, and 21 using HPLC. Replicate results were maintained within 20% for all levels. Mean-measured concentrations were rounded to two significant figures by the study authors. Test organisms were transferred to new test solutions three times a week; however, concentrations were measured on day 0, 7, and 14 only. Solvent (type, percentage, ifused) I N NaOH, concentration Used to enhance solubility ofthe was not reported. test substance. A solvent control group was not included in the study. It is not known ifconcentrations ofNaOH were the same for all levels.

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

Solventconcentration shouldnot exceed0.5 mllLjorstatic tests or 0.1 mllLjor flow-through tests. Recommendedsolvents include dimethylformamide, triethylene glycol, methanol, acetone and ethanol. DEeDrecommends ::::'0.1 mL/L oj solvent. Page 

### 10 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates

-Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


### Remarks Parameter Details

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

### Criteria Lighting l6-hours light, 8-hours dark; light intensity was ca. 590
------------------------------------------- lux (not further specified) Recommended photoperiodis 16 hours lightand

### 8 hours of dark. Recovery ofchemical: 83 to 102% ofnominal Reviewer-calculated from replicate results oftest solutions. Frequency ofmeasurement: Days 0, 7, 14, and 21 LaD: Not reported LOQ: 17.5 mg ai/L Positive control {ifused, indicate Not assessed the chemical and concentrations} Otherparameters, ifany Daphnia were provided food Troutchow (Glencoe) and yeast

Feeding: in their freshly-prepared test (Fleischmann's) were fed to the solutions at each renewal daphnids during culturing; it is interval; however, the type unclear ifthe same food was used and quantity were not during the definitive test. reported. 

### 2. Observations: Table 2: Observations Remarks Parameters Details

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

### Criteria Data endpoints measured - Survival (list) - Days to first brood

----------------------------------------------- - Total no. young produced Recommendedendpoints measured: - No. young/surviving adult

- Survival of first-generation daphnids,

- Growth (terminal length)

-Number of youngproduced per female, - Dry weight (required) and length (optional) of each firstgeneration daphnidalive at the endof the test,

- Observations of other effects or clinicalsigns. 

### Page 11 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates

- Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006


### Remarks Parameters Details

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

### Criteria Observation intervals Survival and reproduction were observed at each renewal period (three times per week). Growth was determined at Day 21 (test termination). Were raw data included? Yes (biological data) Otherobservations, if any None assessed.


### II. RESULTS AND DISCUSSION A. MORTALITY: Following 21 days ofexposure, survival averaged 85% in the control group, and 95, 80, 85, 78,82, and 70% in the mean-measured 32,56, 120,250,480, and 940 mg ai/L treatment levels, respectively. The difference was statistically-reduced (p<0.05) compared to the negative control at the 940 mg ai/L level. The 21-Day ECso

### for immobilization was >940 mg ai/L, the highestmean-measured concentration. The NOAEC for survival was 480 mg ai/L. 

### Table 3: Effect ofIN L5300-20 (Tribenuron-methyl) on Survival, Reproduction, and

G th fD h .



### row 0 ap, nta 

### sp.



### Treatment Mean-Measured Survival, % 

### Days to First No. Young No. Young Mean

(and Nominal) Day 21 Brood per Parent Length, mm Cone. (mg/L) Control 85 9.3 443 116 3.7 (dilution water only) 32 (31) 95 10.4* 320* 85 3.6 56 (62) 80 10.1* 373 115 3.6 120 (125) 85 10.0 340 98 3.6 250 (250) 78 9.7 282* 89 3.6* 480 (500) 82 8.8 126* 36* 3.5* Page 12of29

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -

### Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006
Treatment Mean-Measured Survival, 

### % Days to First No. Young No. Young Mean

; 

### (and Nominal) 

### Day 21 Brood per Parent Length, mm Cone. (mgIL) 

### 940 (1000) 70* 15.6* 6* 2* 3.3* NOAEC, mg ai/L 480 480 250 120 LOAEC, mg ai/L 940 940 480 250 * Significantly different (p<0.05) compared to the control using Dunnett's Test. B. EFFECTS ON REPRODUCTION AND GROWTH: 

### The mean time to first brood release was 9.3 days for the negative control, and 10.4, 10.1, 10.0,9.7,8.8, and 15.6 days for the mean-measured 32,56, 120,250,480, and 940 mg ai/L treatment levels, respectively. Differences were statistically-significant (p<0.05) compared to the control at the 32, 56, and 940 mg ai/L levels. However, the differences observed for the 32 and 56 mg ai/L levels were deemed to be not biologically significant due to the nonlinearity and heteroscedasticity ofthe data. Thus, the NOAEC for time to first brood release was 480 mg ai/L. For the control, and mean-measured 32,56, 120,250,480, and 940 mg ai/L levels, the total numbers ofyoung produced were 443,320,373,340,282, 126, and 6, respectively, and the number ofoffspring per surviving parent averaged 116, 85, 115, 98, 89, 36, and 2, respectively. For total offspring, differences were statistically-significant (p<0.05) compared to the control at the 32, 250, 480, and 940 mg ai/L levels. However, when the data were corrected for surviving parent, differences remained statistically-significantonly at the 480 and 940 mg ai/L levels. Thus, the NOAEC for offspring production was 250 mg ai/L. It 

### was reported that the observed ECso

### forreproduction (calculated using failure to produce eggs as the observation criterion) was 

### ca. 900 mg ai/L. No males, ephippia, or immobilizedyoung were observed in the control group or any treatment level during the study, and no eggs were observed in the bottom ofany test vessel. Growth, measured as length at study termination, was the most sensitive endpoint. Mean lengths were 3.7,3.6,3.6,3.6,3.6,3.5, and 3.3 mm for the control, and mean-measured 32, 56, 120,250,480, and 940 mg ai/L levels, respectively. Differences were statisticallysignificant (p<0.05) comparedto the control at the 250, 480, and 940 mg ai/L levels, respectively. The NOAEC for growth was 120 mg ai/L. 

### C. REPORTED STATISTICS: 

### Data that were statistically analyzed included organism survival, time to first brood release, Page 13 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 total number ofoffspring, number ofoffspring produced per surviving adult and total body length. Survival data were analyzed using Fisher's Exact test and the Cochran-Armitage trend test. Reproductive and growth endpoints were analyzed using a one-way analysis of variance (ANOYA) followed by Dunnett's procedure. During this study, no concentration caused 2:50% immobilization; therefore, the 21-day ECso was empirically estimated to be greater than the highest tested concentration. The 21-day ECso for reproduction was calculated by Probit analysis using failure to produce eggs as the observation criterion. All results were reported in terms ofmean-measured concentrations. Parental Survival ECso: > 940 mg ai/L NOAEC: 480 mg ai/L LOAEC: 940 mg ai/L Time to First Brood NOAEC: 480 mg ai/L LOAEC: 940 mg ai/L Offspring/surviving adult ECso: ca. 

### 900 mg ai/L Probit slope: Not reported NOAEC: 250 mg ai/L LOAEC: 480 mg ai/L Total Length NOAEC: 120 mg ai/L LOAEC: 250 mg ai/L 95% C.l.: N/A

95% C.l.: Not reported 95% C.I.: Not reported



### D. VERIFICATION OF STATISTICAL RESULTS: Statistical Method(s): The reviewer determined the NOAEC for immobility using Fisher's Exact Test. Reproduction (time to first brood, total offspring, offspring produced/day per surviving adult) and growth (length) endpoint data were determined to be normally distributed using the Chi-square test and variances were confirmed to be homogeneous using Levene's test. The NOAEC and LOAEC for these endpoints were determined using ANOYA, followed by Dunnett's test. These analyses were conducted using Toxstat 3.5. The reviewer determinedthe 21-day ECsofor reproductive endpoints (total offspring production and offspring produced/day per surviving adult) using the probitmethod via Nuthatch statistical software. Parental Survival ECso: > 

### 940 mg ai/L 95% C.l.: N/A Page 14of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates

-

### Daphnia sp. PMRA SubmissionNumber {........} EPA MRID No. 47643006 NOAEC: 940 mg ai/L LOAEC: >940 mg ai/L Time to First Brood NOAEC: 480 mg ai/L LOAEC: 940 mg ai/L Total Offspring produced ECso: 

### 380 mg ai/L Probit slope: 5.07±0.724 NOAEC: <32 mg ai/L LOAEC: 32 mg ai/L 95% 

## c.r. 330-440 mg ai/L

Offspring/day per surviving adult ECso: 

### 410 mg ai/L 95% C.l.: 350-470 mg ai/L Probit slope: 5.64±0.920 NOAEC: 120 mg ai/L LOAEC: 250 mg ai/L Total Length NOAEC: <32 mg ai/L LOAEC: 32 mg ai/L 

### E. STUDY DEFICIENCIES: This study is considered scientificallyunsound, as the dissolved oxygen was <60% (equivalentto 5.5 mg/L at 20°C; reviewer-calculated) for an unspecifiedperiod oftime at all levels during the study. In addition, the source ofthe dilution (fish culture water) could have contained ammonia in the water, which was neithermeasured nor reported in the water quality measurements. The combination ofthese multiple stressors may have impacted the test organisms, and after analysis ofthe data the reviewerconcludedthat aNOAEC could not be determined for this study due to significant (p<0.05) suggestive treatment effects on total offspring production and adult length at the lowesttreatment level. Furthermore, the following notable deviations from OPPTS 850.1300 guidance were observed: dissolved oxygen in the test vessels ofaged solutions was <60%; and although a 1 N NaOH solution was used to enhance solubility ofthe test substance, a solvent control group was not included in the study. Additionally, it was unknown what the concentrations ofNaOH were and if those concentrations were the same for all treatment levels. 

### F. REVIEWER'S COMMENTS: The reviewer's conclusions differed from the study author's. While both analyses detected significantreductions in total offspring production at the lowest treatment level, the study Page 15 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber {........} EPA MRID No. 47643006 author dismissedthe effect because overall it was not dose-dependent. The study author did not note a similar reduction for adult length at the lowest level, which the reviewer's analysis detected. Additionally, the reviewer's analysis detected a lower NOAEC for offspring produced/day per surviving adult than the study author's analysis, but the reviewer's analysis did not detect any effect on adult immobility. The reviewer's more conservative conclusions regarding reproduction and length are presented in the Executive Summary and Conclusions sections. All OPPTS 850.1300 validity requirements were fulfilled. Specifically, control mortality was 15% (no more than 20% permissible), control daphnids each produced an average of 116 young (minimum of60 required), and no ephippia were produced by control daphnids. It

### was reported that a 1 N sodium hydroxide (NaOH) solution was used to bring the pH ofthe stock solutions up to ca. 

### 9 before testing in order to increase solubility ofthe test substance. No further details were provided, so it was unknown ifconcentrations ofNaOH were the same for all levels. A solvent control group was not included in the study design. Test samples collected for analytical verification were stored refrigerated prior to analysis. Aliquots were analyzed for Tribenuron-methyl either directly or following dilution with water using high-performance liquid chromatography (HPLC) with UV detection (230 nm). The experimental phase ofthe definitive study was conducted from February 22 to March 15, 1989. 

### G. CONCLUSIONS: This study is scientifically unsound

### and is thus not acceptable. The dissolved oxygen was <60% (equivalentto 5.5 mg/L at 20°C; reviewer-calculated) for an unspecified period of time at all levels during the study, and the source ofthe dilution water (fish culture water) could have contained ammonia. The combination ofthese multiple stressors may have negatively impactedthe test organisms, and the resulting analysis ofthe data indicated that a NOAEC could not be determined «32 mg ai/L, the lowest test concentration). Based upon a treatment-relatedreduction in offspring/day per surviving adult and in terminal lengths of surviving daphnia at the 32 mg ai/L treatment level (the most sensitive endpoints), the overall NOAEC for the study could not be determined. A treatment-related reduction in offspring production/day per surviving adult was also indicated at the 250, 480 and 940 mg ai/L levels, and atreatment-related delay offirst brood release was indicated at the 940 mg ai/L level. The ECsovalues for immobility and reproduction (total offspring) were >940 and 380 mg ai/L, respectively. Based on the major deviations that were noted above, the results from this study are not reliable, and are not acceptable for use. Therefore, this study is classified as 

### invalid. 21-day ECso(mortality/immobility): >940 mg ai/L 21-day ECso (reproduction): 380 mg ai/L Page 16of29 95% C.L: N/A 95% C.L: 330-440 mg ai/L

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp, 

### PMRA SubmissionNumber { } EPA MRID No. 47643006 Slope: 5.07±O.724 NOAEC: <32 mg ai/L LOAEC: 32 mg ai/L Endpoints affected: Survival, time to first brood release, offspring production, and growth (length) Most sensitive endpoint(s): Total offspring production and growth (length) 

### III. REFERENCES: Hutton, D.G. 1989. "Short-term Study Protocol Sheet", MR-4581-665, Haskell Number 15,527, February 20, 1989. "Chronic Toxicity Test on Compounds with Daphnia magna", 

### Haskell laboratory Aquatic Test SOP-T23. OECD. 1984. "OECD Guideline for Testing ofChemicals: 202",4 April 84. Rexrode, M, and T.M. Armitage. 1986. "Standard Evaluation Procedure, Daphnia magna 

### LifeCycle (21-Day Renewal) chronic Toxicity Test". EPA 540/9-86-141, U.S. EPA, Office of Pesticide programs, Washington, DC, July 1986. Hutton, D.G. 1984. "48-HourEC50 to Daphnia magna", 

### Haskell Laboratory Report No. 461- 84, Revised November 15, 1984. Hutton, D.G. "Static Acute 48-HourEC50 ofIN L5300-20 to FedDaphnia magna", 

### Haskell Laboratory Report to be issued. Siegel, S. 1956. Nonparametric Statisticsfor the BehavioralSciences. 

### McGraw-Hill, New York, pp. 96-104. Finney, D.J. 1971. Probit Analysis, 3rd 

### Ed. Cambridge University Press. 333 pp. Steel, R.G.D., and J.H. Torrie. 1980. Principles andProcedures a/Statistics. 

### 2nd

### Edition, McGraw-Hill, New York. Milliken, G.A., and D.E. Johnson. 1984. Analysis

## 0/ Messy Data, 

### Volume I, Lifetime Learning Publications, Belmont, CA. Haskell Laboratory MR-5659.

Page 17 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA 

### MRID No. 47643006 APPENDIX I. OUTPUT OF REVIEWER'S STATISTICAL VERIFICATION: Summary of Fisher's Exact Tests NUMBER NUMBER SIG GROUP IDENTIFICATION EXPOSED DEAD 0.05

------------------------- ----------- ----------- CONTROL 40 6 1 32 40 2 2 56 40 8 3 120 40 6 4 250 40 9 5 480 40 7 6 940 40 12
Title: File: First Day of Brood 3006b Transform: Chi-Square Test for Normality NO TRANSFORMATION

INTERVAL <-1.5 Actual and Expected Frequencies -1.5 to <-0.5 -0.5 to 0.5 >0.5 to 1.5 >1. 5 EXPECTED OBSERVED 4.6900 1 16.9400 20 26.7400 30 16.9400 16 4.6900 3 Chi-Square 

### = 4.5145 Critical Chi-Square 

### = 13.277 9.488 (p-value 

### = 0.3408) (alpha 

### = 0.01 , df 

### = 4) (alpha 

### = 0.05 , df 

### = 4) Data PASS normality test (alpha = 0.01). Continue analysis. Title: File: First Day of Brood 3006b Transform: NO TRANSFORMATION

Levene's Test for Homogeneity of Variance ANOVA Table SOURCE Between Within (Error) Total DF
6 63 69 SS 53.0857 106.0000 159.0857 MS 8.8476 1.6825 F 5.2585

Critical F 3.1028 (alpha 2.2464 (alpha 0.01, df 0.05, df 6,63) 6,63) (p-value = 0.0002)

Since F > Critical F REJECT Ho: All equal (alpha



### Page 18 of29 0.01)

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006 Title: File: First Day of Brood 3006b Transform: ANOVA Table NO TRANSFORMATION

SOURCE Between Within (Error) Total DF 6 63 69 SS 313.7714 283.5000 597.2714 MS 52.2952

4.5000 F 11.6212

Critical F 

### = 3.1028 

### = 2.2464 Since F > Critical F (alpha 

### = 0.01, df 

### = 6,63) (alpha 

### = 0.05, df 

### = 6,63) REJECT Ho: All equal (alpha (p-value = 0.0000) 0.05) Title: File: First Day of Brood 3006b Dunnett's Test Transform: TABLE 1 OF 2 NO TRANSFORMATION Ho:Control>Treatment TRANSFORMED MEAN CALCULATED IN SIG GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT 0.05

-------------------- ----------- ------------------ ------ 1 Control 9.3000 9.3000 2 32 10.4000 10.4000 1.1595 3 56 10.1000 10.1000 0.8433 4 120 10.0000 10.0000 0.7379 5 250 9.7000 9.7000 0.4216 6 480 8.8000 8.8000 -0.5270 7 940 15.6000 15.6000 6.6408 

### * Dunnett critical value = 2.3400 (1 Tailed, alpha 

### = 0.05, df [used] 6,60) (Actual df 

### = 6,63) Title: File: First Day of Brood 3006b Dunnett's Test Transform: TABLE 2 OF 2 NO TRANSFORMATION Ho:Control>Treatment

---------------------------------------------------------------------------- NUM OF MIN SIG DIFF % OF DIFFERENCE GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

-------------------- ------- ---------------- ------- ------------ 1 Control 10 2 32 10 2.2199 23.9 1.1000 3 56 10 2.2199 23.9 0.8000 4 120 10 2.2199 23.9 0.7000 5 250 10 2.2199 23.9 0.4000 6 480 10 2.2199 23.9 -0.5000 7 940 10 2.2199 23.9 6.3000 

### Page 19of29

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA Submission Number { } EPA MRID No. 47643006

Title: File: First Day of Brood 3006b William's Test Transform:

- TABLE 1 OF 2 NO TRANSFORMATION Ho: Control>Treatment ORIGINAL TRANSFORMED ISOTONIZED GROUP IDENTIFICATION N MEAN MEAN MEAN

------ -------------------- ----------- ----------- ----------- 1 Control 10 9.3000 9.3000 9.3000 2 32 10 10.4000 10.4000 9.8000 3 56 10 10.1000 10.1000 9.8000 4 120 10 10.0000 10.0000 9.8000 5 250 10 9.7000 9.7000 9.8000 6 480 10 8.8000 8.8000 9.8000 7 940 10 15.6000 15.6000 15.6000
Title: File: First Day of Brood 3006b William's Test Transform:

- TABLE 2 OF 2 NO TRANSFORMATION Ho: Control>Treatment

---------------------------------------------------------------------------- COMPARED CALC. SIG TABLE DEGREES OF IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM USED

-------------------- ----------- ----------- ----------- ------------ Control 9.3000 32 9.8000 0.5270 1.6700 k= 1, v=60 56 9.8000 0.5270 1.7500 k= 2, v=60 120 9.8000 0.5270 1.7700 k= 3, v=60 250 9.8000 0.5270 1.7800 k= 4, v=60 480 9.8000 0.5270 1.7900 k= 5, v=60 940 15.6000 6.6408 

### * 1.7900 k= 6, v=60

s = 2.1213

WARNING: Procedure has used isotonized means which differ from original (transformed) means. Title: File: Total Offspring produced 30060 Transform: Chi-Square Test for Normality NO TRANSFORMATION

INTERVAL <-1. 5 Actual and Expected Frequencies -1.5 to <-0.5 -0.5 to 0.5 >0.5 to 1.5 >1. 5 EXPECTED OBSERVED 4.6900 1 16.9400 23 26.7400 25 16.9400 14 4.6900 7 Chi-Square = 6.8323 Critical Chi-Square = 13.277 9.488 (p-value = 0.1450) (alpha = 0.01 , df = 4) (alpha = 0.05 , df = 4) Data PASS normality test (alpha = 0.01). Continue analysis. 

### Page 20 of29

### Data Evaluation Record on the Chronic Toxicity ofIN L5300-20 

### (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia sp. 

### PMRA SubmissionNumber { } EPA MRID No. 47643006

Title: File: Total Offspring produced 30060 Transform:

Levene's Test for Homogeneity ANOVA Table NO TRANSFORMATION of Variance

SOURCE DF SS MS F Between Within (Error) 6 63 65694.0857 221621.9000 10949.0143 3517.8079 3.1125
Total 69 287315.9857 (p-value = 0.0098)

Critical F 

### = 3.1028 

### = 2.2464 Since F > Critical F (alpha = 0.01, df = 6,63) (alpha 

### = 0.05, df 

### = 6,63) REJECT Ho: All equal (alpha 0.01) Title: File: Total Offspring produced 30060 Transform: ANOVA Table NO TRANSFORMATION
SOURCE DF SS MS F Between Within (Error) 6 63 1388497.2857 572605.3000 231416.2143 9088.9730 25.4612
Total 69 1961102.5857 (p-value 

### = 0.0000) Critical F 

### = 3.1028 

### = 2.2464 Since F > Critical F (alpha 

### = 0.01, df 

### = 6,63) (alpha 

### = 0.05, df 

### = 6,63) REJECT Ho: All equal (alpha 0.05)
NO TRANSFORMATION Ho:Control<Treatment Transform: TABLE 1 OF 2 Total Offspring produced 30060 Dunnett's Test Title: File:
TRANSFORMED MEAN CALCULATED IN SIG GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT 0.05

-------------------- ----------- ------------------ ------ 1 Control 442.8000 442.8000 2 32 319.9000 319.9000 2.8826 

### * 3 56 373.1000 373.1000 1.6348 4 120 339.9000 339.9000 2.4135 

### * 5 250 282.0000 282.0000 3.7715 

### * 6 480 125.5000 125.5000 7.4421 

### * 7 940 5.5000 5.5000 10.2567 

### * Dunnett critical value 

### = 2.3400 (I Tailed, alpha 

### = 0.05, df [used] 6,60) (Actual df 

### = 6,63)


### Page 21 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia sp. PMRA Submission Number { } EPA MRID No. 47643006

Title: File: Total Offspring produced 30060 Transform: Dunnett's Test TABLE 2 OF 2 NO TRANSFORMATION Ho:Control<Treatment

---------------------------------------------------------------------------- NUM OF MIN SIG DIFF % OF DIFFERENCE GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

-------------------- ------- ---------------- ------- ------------ 1 Control 10 2 32 10 99.7673 22.5 122.9000 3 56 10 99.7673 22.5 69.7000 4 120 10 99.7673 22.5 102.9000 5 250 10 99.7673 22.5 160.8000 6 480 10 99.7673 22.5 317.3000 7 940 10 99.7673 22.5 437.3000

Title: File: Total Offspring produced 30060 William's Test Transform: - TABLE 1 OF 2 NO TRANSFORMATION Ho: Control<Treatment

---------------------------------------------------------------------------- ORIGINAL TRANSFORMED ISOTONIZED GROUP IDENTIFICATION N MEAN MEAN MEAN

------ -------------------- ----------- ----------- ----------- 1 Control 10 442.8000 442.8000 442.8000 2 32 10 319.9000 319.9000 346.5000 3 56 10 373.1000 373.1000 346.5000 4 120 10 339.9000 339.9000 339.9000 5 250 10 282.0000 282.0000 282.0000 6 480 10 125.5000 125.5000 125.5000 7 940 10 5.5000 5.5000 5.5000

Title: File: Total Offspring produced 30060 William's Test Transform: - TABLE 2 OF 2 NO TRANSFORMATION Ho: Control<Treatment

---------------------------------------------------------------------------- COMPARED CALC. SIG TABLE DEGREES OF IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM USED

-------------------- ----------- ----------- ----------- ------------ Control 442.8000 32 346.5000 2.2587 

### * 1.6700 k= 1, v=60 56 346.5000 2.2587 

### * 1.7500 k= 2, v=60 120 339.9000 2.4135 

### * 1.7700 k= 3, v=60 250 282.0000 3.7715 

### * 1.7800 k= 4, v=60 480 125.5000 7.4421 

### * 1.7900 k= 5, v=60 940 5.5000 10.2567 

### * 1.7900 k= 6, v=60

s = 95.3361

WARNING: Procedure has used isotonized means which differ from original (transformed) means. 

### Page 22 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -Daphnia 

### sp. PMRA Submission Number { } EPA MRID No. 47643006
30060 : Total Offspring produced Williams Test [One-Sided Test for Decrease, alpha = 0.050000 ] Dose Isotone Means T-bar P-value Significance



### o 32 56 120 250 480 940 443 347 347 340 282 126 5.5 2.259 2.259 2.413 3.771 7.442 10.26 0.014 0.016 0.011 <0.005 <0.005 <0.005 

### * * * 

### * * 

### * "*"=Significant; "N.S."=Not Significant. Estimates of EC% Parameter Estimate 95% Bounds Std.Err. Lower Bound Lower Upper /Estimate EC5 1.8E+02 1.3E+02 2.5E+02 0.072 0.72 EC10 2.1E+02 1.6E+02 2.8E+02 0.062 0.75 EC25 2.8E+02 2.3E+02 3.5E+02 0.047 0.81 EC50 3.8E+02 3.3E+02 4.4E+02 0.032 0.86 Slope = 5.07 Std.Err. = 0.724 Goodness of fit: p = 0.055 based on DF= 4.0 63. 30060 : Total Offspring produced Observed vs. Predicted Treatment Group Means

------------------------------------------------------------------------ Dose #Reps. Obs. Pred. Obs. Pred. %Change Mean Mean -Pred. %Control 0.00 10.0 443. 367. 76.1 100. 0.00 32.0 10.0 320. 367. -46.8 100. 2.42e-06 56.0 10.0 373. 367. 6.40 100. 0.00119 120. 10.0 340. 365. -24.8 99.5 0.542 250. 10.0 282. 302. -20.4 82.5 17.5 480. 10.0 126. 113. 12.7 30.8 69.2 940. 10.0 5.50 8.71 -3.21 2.37 97.6



### Page 23 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA Submission Number { } EPA MRID No. 47643006 Title: File: Young/Day/Surviving Parent 3006y Transform: Chi-Square Test for Normality NO TRANSFORMATION

INTERVAL <-1. 5 Actual and Expected Frequencies -1.5 to <-0.5 -0.5 to 0.5 >0.5 to 1.5 >1. 5 EXPECTED OBSERVED 4.6900 2 16.9400 23 26.7400 28 16.9400 9 4.6900 8 Chi-Square 

### = 9.8278 Critical Chi-Square = 13.277 9.488 (p-value 

### = 0.0434) (alpha 

### = 0.01 , df 

### = 4) (alpha = 0.05 , df = 4) Data PASS normality test (alpha = 0.01). Continue analysis. Title: File: Young/Day/Surviving Parent 3006y Transform:

Levene's Test for Homogeneity ANOVA Table NO TRANSFORMATION of Variance

SOURCE Between Within (Error) Total DF 6 63 69 SS 7134.5337 23405.2440 30539.7777 MS 1189.0890

371.5118 F 3.2007

(p-value 

### = 0.0083) Critical F 

### = 3.1028

= 2.2464 Since F > Critical F (alpha 

### = 0.01, df 

### = 6(63) (alpha 

### = 0.05, df 

### = 6,63) REJECT Ho: All equal (alpha 0.01) Title: File: Young/Day/Surviving Parent 3006y Transform: ANOVA Table NO TRANSFORMATION

SOURCE Between Within (Error) Total DF
6 63 69 SS 109122.2480

58272.0840 167394.3320 MS 18187.0413 924.9537 F 19.6627

(p-value 

### = 0.0000) Critical F = 3.1028 

### = 2.2464 Since F > Critical F (alpha 

### = 0.01, df 

### = 6(63) (alpha 

### = 0.05, df 

### = 6,63) REJECT Ho: All equal (alpha



### Page 24 of29 0.05)

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006

Transform: TABLE 1 OF 2 Title: File: Young/Day/Surviving Parent 3006y Dunnett's Test NO TRANSFORMATION Ho:Control<Treatment TRANSFORMED MEAN CALCULATED IN SIG GROUP IDENTIFICATION MEAN ORIGINAL UNITS T STAT 0.05

-------------------- ----------- ------------------ ------ 1 Control 115.5000 115.5000 2 32 84.5000 84.5000 2.2792 3 56 114.9000 114.9000 0.0441 4 120 97.9000 97.9000 1.2940 5 250 89.2000 89.2000 1.9337 6 480 35.7000 35.7000 5.8672 

### * 7 940 1.8600 1.8600 8.3552 

### * Dunnett critical value = 2.3400 (1 Tailed, alpha = 0.05, df [used] =6,60) (Actual df = 6,63)

Transform: TABLE 2 OF 2 Title: File: Young/Day/Surviving Parent 3006y Dunnett's Test NO TRANSFORMATION Ho:Control<Treatment

---------------------------------------------------------------------------- NUM OF MIN SIG DIFF % OF DIFFERENCE GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

-------------------- ------- ---------------- ------- ------------ 1 Control 10 2 32 10 31.8266 27.6 31.0000 3 56 10 31.8266 27.6 0.6000 4 120 10 31.8266 27.6 17.6000 5 250 10 31.8266 27.6 26.3000 6 480 10 31.8266 27.6 79.8000 7 940 10 31.8266 27.6 113.6400

Title: File: Young/Day/Surviving Parent 3006y Transform: William's Test - TABLE 1 OF 2 NO TRANSFORMATION Ho: Control<Treatment ORIGINAL TRANSFORMED ISOTONIZED GROUP IDENTIFICATION N MEAN MEAN MEAN

------ -------------------- ----------- ----------- ----------- 1 Control 10 115.5000 115.5000 115.5000 2 32 10 84.5000 84.5000 99.7000 3 56 10 114.9000 114.9000 99.7000 4 120 10 97.9000 97.9000 97.9000 5 250 10 89.2000 89.2000 89.2000 6 480 10 35.7000 35.7000 35.7000 7 940 10 1.8600 1.8600 1.8600



### Page 25 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to FreshwaterInvertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006

Title: File: Young/Day/Surviving Parent 3006y Transform: William's Test - TABLE 2 OF 2 NO TRANSFORMATION Ho: Control<Treatment COMPARED CALC. SIG TABLE DEGREES OF IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM USED

-------------------- ----------- ----------- ----------- ------------ Control 115.5000 32 99.7000 1. 1617 1.6700 k= 1, v=60 56 99.7000 1. 1617 1.7500 k= 2, v=60 120 97.9000 1.2940 1.7700 k= 3, v=60 250 89.2000 1.9337 * 1.7800 k= 4, v=60 480 35.7000 5.8672 * 1.7900 k= 5, v=60 940 1.8600 8.3552 * 1.7900 k= 6, v=60

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

s = 30.4131

WARNING: Procedure has used isotonized means which differ from original (transformed) means. Williams Test [One-Sided Test for Decrease, alpha = Dose Isotone T-bar P-value Means 0.050000 ] Significance

0 116 32 99.7 1.162 N.S. 56 99.7 1.162 N.S. 120 97.9 1.294 N.S. 250 89.2 1.934 0.036 * 480 35.7 5.867 <0.005 * 940 1. 86 8.355 <0.005 *

"*"=Significant; "N.S."=Not Significant. Estimates of EC% Parameter EC5 EC10 EC25 EC50 Estimate 2.1E+02 2.4E+02 3.1E+02 4.1E+02 95% Bounds Lower Upper 1.5E+02 2.9E+02 1.8E+02 3.2E+02 2.5E+02 3.8E+02 3.SE+02 4.7E+02 Std. Err. 0.073 0.063 0.048 0.033 Lower Bound /Estimate 0.72 0.75 0.80 0.86 Slope = 5.64 Std.Err. = 0.920 Goodness of fit: p = 0.17 based on DF= 4.0 63. 3006Y : Young/Day/Surviving Parent Observed vs. Predicted Treatment Group Means 

### Page 26 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA 

### MRID No. 47643006
Dose #Reps. Obs. Pred. Obs. Pred. %Change Mean Mean -Pred. %Control 0.00 10.0 116. 103. 12.5 100. 0.00 32.0 10.0 84.5 103. -18.5 100. 2.43e-08 56.0 10.0 115. 103. 11.9 100. 6.11e-05 120. 10.0 97.9 103. -4.92 99.9 0.142 250. 10.0 89.2 90.8 -1.63 88.2 11. 8 480. 10.0 35.7 35.0 0.729 34.0 66.0 940. 10.0 1. 86 2.02 -0.165 1. 97 98.0 Title: Total length File: 30061 Transform: NO TRANSFORMATION Chi-Square Test for Normality

INTERVAL <-1.5 Actual and Expected Frequencies -1.5 to <-0.5 -0.5 to 0.5 >0.5 to 1.5 >1. 5 EXPECTED OBSERVED 15.4100 18 55.6600 52 87.8600 83 55.6600 63 15.4100 14 Chi-Square 

### = 2.0418 (p-value 

### = 0.7281) Critical Chi-Square 13.277 9.488 (alpha 

### = 0.01 , df 

### = 4) (alpha 

### = 0.05 , df 

### = 4) Data PASS normality test (alpha 

### = 0.01). Continue analysis. Title: File: Total length 30061 Transform: NO TRANSFORMATION

Levene's Test for Homogeneity of Variance ANOVA Table SOURCE Between Within (Error) Total DF 6 223 229 SS 0.0994 2.8039 2.9033 MS 0.0166 0.0126 F 1.3178

(p-value 

### = 0.2501) Critical F 

### = 2.8837

= 2.13 94 Since F < Critical F (alpha 

### = 0.01, df 

### = 6,223) (alpha 

### = 0.05, df 

### = 6,223) FAIL TO REJECT Ho: All equal (alpha



### Page 27 of29 0.01)

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -

### Daphnia sp, PMRA SubmissionNumber { } EPA MRID No. 47643006

Title: File: Total length 30061 Transform: ANOVA Table NO TRANSFORMATION

SOURCE DF SS MS F Between Within (Error) Total 6 223 229 3.1652 6.9435 10.1087 0.5275 0.0311 16.9423

Critical F = 2.8837
= 2.1394 Since F > Critical F (alpha = 0.01, df 

### = 6,223) (alpha 

### = 0.05, df 

### = 6,223) REJECT Ho: All equal (alpha (p-value = 0.0000) 0.05) Title: File: Total length 30061 Bonferroni t-Test Transform: TABLE 1 OF 2 NO TRANSFORMATION Ho: Control<Treatment TRANSFORMED MEAN CALCULATED IN SIG GROUP IDENTIFICATION MEAN ORIGINAL UNITS t STAT 0.05

-------------------- ----------- ------------------ ------ 1 Control 3.7118 3.7118 2 32 3.6000 3.6000 2.6831 * 3 56 3.6344 3.6344 1.7807 4 120 3.6353 3.6353 1.7868 5 250 3.5516 3.5516 3.6547 * 6 480 3.5485 3.5485 3.7866 * 7 940 3.2929 3.2929 9.3026 * Bonferroni t critical value = 2.4122 (1 Tailed, alpha 

### = 0.05, df 

### = 6,223) Title: File: Total length 30061 Bonferroni t-Test Transform: TABLE 2 OF 2 NO TRANSFORMATION Ho: Control<Treatment

---------------------------------------------------------------------------- NUM OF MIN SIG DIFF % OF DIFFERENCE GROUP IDENTIFICATION REPS (IN ORIG. UNITS) CONTROL FROM CONTROL

-------------------- ------- ---------------- ------- ------------ 1 Control 34 2 32 38 0.1005 2.7 0.1118 3 56 32 0.1048 2.8 0.0774 4 120 34 0.1032 2.8 0.0765 5 250 31 0.1057 2.8 0.1602 6 480 33 0.1040 2.8 0.1633 7 940 28 0.1086 2.9 0.4189



### Page 28 of29

### Data Evaluation Record on the Chronic Toxicity of IN L5300-20 (Tribenuron-methyl) to Freshwater Invertebrates -Daphnia 

### sp. PMRA SubmissionNumber { } EPA MRID No. 47643006

Title: File: Total length 30061 William's Test Transform:

- TABLE 1 OF 2 NO TRANSFORMATION Ho: Control<Treatment ORIGINAL TRANSFORMED ISOTONIZED GROUP IDENTIFICATION N MEAN MEAN MEAN

------**------~-------------** ----------- ----------- ----------- 1 Control 34 3.7118 3.7118 3.7118 2 32 38 3.6000 3.6000 3.6221 3 56 32 3.6344 3.6344 3.6221 4 120 34 3.6353 3.6353 3.6221 5 250 31 3.5516 3.5516 3.5516 6 480 33 3.5485 3.5485 3.5485 7 940 28 3.2929 3.2929 3.2929
Title: File: Total length 30061 William's Test Transform:

- TABLE 2 OF 2 NO TRANSFORMATION Ho: Control<Treatment COMPARED CALC. SIG TABLE DEGREES OF IDENTIFICATION MEANS WILLIAMS 0.05 WILLIAMS FREEDOM USED

-------------------- ----------- ----------- ----------- ------------ Control 3.7118 32 3.6221 2.1522 

### * 1.6600 k= 1, v=120 56 3.6221 2.0628 

### * 1.7300 k= 2, v=120 120 3.6221 2.0948 

### * 1.7500 k= 3, v=120 250 3.5516 3.6547 

### * 1.7700 k= 4, v=120 480 3.5485 3.7866 

### * 1.7700 k= 5, v=120 940 3.2929 9.3026 

### * 1.7800 k= 6, v=120

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

s = 0.1765

WARNING: Procedure has used isotonized means which differ from original (transformed) means.



### Page 29 of29